zearalenone and deoxynivalenol

Mycotoxin, as one of the most common pollutants in foodstuffs, poses great threat to food security and human health. Specifically, deoxynivalenol (DON) and zearalenone (ZEN)-two mycotoxin contaminants with considerable toxicity widely existing in food products-have aroused broad public concerns. Adding to this picture, modified forms of DON and ZEN, have emerged as another potential environmental and health threat, owing to their higher re-transformation rate into parent mycotoxins inducing accumulation of mycotoxin in humans and animals. Given this, a better understanding of the toxicity of modified mycotoxins is urgently needed. Moreover, the lack of toxicity data means a proper risk assessment of modified mycotoxins remains challenging. To better evaluate the toxicity of modified DON and ZEN, we have reviewed the relationship between their structures and toxicities. The toxicity mechanisms behind modified DON and ZEN have also been discussed; briefly, these involve acute, subacute, chronic, and combined toxicities. In addition, this review also addresses the global occurrence of modified DON and ZEN, and summarizes novel methods-including in silico analysis and implementation of relative potency factors-for risk assessment of modified DON and ZEN. Finally, the health risk assessment of modified DON and ZEN has also been discussed comprehensively.

Topics: Animals; Food Contamination; Humans; Mycotoxins; Risk Assessment; Trichothecenes; Zearalenone

Mycotoxin contamination of cereals used for feed can cause intoxication, especially in farm animals; therefore, efficient analytical tools for the qualitative and quantitative analysis of toxic fungal metabolites in feed are required. Current trends in food/feed analysis are focusing on the application of biosensor technologies that offer fast and highly selective and sensitive detection with minimal sample treatment and reagents required. The article presents an overview of the recent progress of the development of biosensors for deoxynivalenol and zearalenone determination in cereals and feed. Novel biosensitive materials and highly sensitive detection methods applied for the sensors and the application of these sensors to food/feed products, the limit, and the time of detection are discussed.

Topics: Animal Feed; Animals; Biosensing Techniques; Edible Grain; Food Analysis; Food Contamination; Fungi; Fusarium; Mycotoxins; Quality Control; Trichothecenes; Zearalenone

Deoxynivalenol (DON) and Zearalenone (ZEN) are two commonly co-occurring mycotoxins produced by members of the genus

Topics: Animals; Drug Interactions; Food Chain; Food Contamination; Humans; Trichothecenes; Zearalenone

Human exposure to mycotoxins occurs mostly through dietary intake, although exposure through dermal and inhalation routes has also been shown. Depending on the type of mycotoxins, the applied dose and duration of exposure, a particular toxin can cause either chronic or acute illnesses such as kidney failure and cancer. Thus, understanding the biotransformation of mycotoxins and identification of reliable biomarkers in the human body is important for accurate risk assessment of mycotoxin exposure. This review provides a comprehensive overview of worldwide aflatoxins, fumonisins, ochratoxin, zearalenone and deoxynivalenol mycotoxin biomonitoring studies reported in the last 18 years. The studies performed in Africa, Europe, Asia and America are based on the measurement of a limited number of mycotoxin biomarkers and do not provide a comprehensive risk assessment of the mycotoxin exposure. Although the findings represent a small segment of a much larger health risk of mycotoxins exposure, it is acknowledged that a multianalyte approach covering bioconjugated and other metabolites of most often occurring mycotoxins would better reflect the extent of the global exposure problems with these highly toxic compounds.

Topics: Aflatoxins; Africa; Biomarkers; Body Fluids; Dietary Exposure; Environmental Monitoring; Europe; Fumonisins; Humans; Ochratoxins; Risk Assessment; Trichothecenes; Zearalenone

This study aimed to estimate the prevalence and concentration of total aflatoxin (TAF) ochratoxin A (OTA), zearalenone (ZEN) and deoxynivalenol (DON) in bread, cornflakes, breakfast cereals and pasta-based products through meta-analysis. The required databases including (PubMed and Scopus databases) were investigated to collect data on the concentration and prevalence of mentioned mycotoxins in cereal-based products. Among 2461 explored articles in identification step, 38 articles with 9627 samples were included in the conducted meta-analysis. The prevalence and concentration of studied mycotoxins varied with the cereal-based food studied. In this context, the overall rank order of mycotoxins prevalence in the cereal foods was OTA > DON > ZEN > TAF > 15-ADON > 3-ADON. Also, the overall rank order of mycotoxins based on concentration in the cereal foods investigated was DON > ZEN > 15-ADON > OTA > 3-ADON > TAF. The findings of this meta-analysis may be useful for the building of risk assessment models aiming to derive data for the development of specific actions to reduce the exposure to OTA, ZEN, TAF, and DON through the consumption of the cereal-based products.

Topics: Aflatoxins; Edible Grain; Food Contamination; Ochratoxins; Prevalence; Trichothecenes; Zearalenone

The study aimed to perform a meta-analysis on the fate of ochratoxin A (OTA), zearalenone (ZEN), deoxynivalenol (DON) and total aflatoxin (TAF) during steps of bread and pasta-based products processing. A total of twenty and eight articles (549 data) collected from 1983 through June 2017 were included. Some of the investigated processing such as milling and fermentation caused an increase in the concentration of DON and TAF; although they reduce the concentration of ZEN and OTA. Also, heat processing (cooking) decrease the DON, OTA, and TAF and increase the concentration of ZEN in bread. Cooking reduces the concentration of DON and ZEN in the biscuit. Cooking of pasta reduces the content of DON; however, it increases the concentration of TFA. The findings showed that the mycotoxins and their fate were influenced differently by the unit operations steps involved in the preparation of the different cereal-based products.

Topics: Aflatoxins; Edible Grain; Food Contamination; Mycotoxins; Ochratoxins; Trichothecenes; Zearalenone

There is an increasing awareness of the deleterious effects attributed to mycotoxins during their fate within the gut, particularly for deoxynivalenol (DON), zearalenone (ZEN), ochratoxin A (OTA), fumonisin B1 (FB1), aflatoxin B1 (AFB1), and patulin (PAT). Evidence indicates that disruption of the epithelial barrier is well established. However, intestinal barrier function on its luminal side involves two other partners, mucus and microbiota, which have rarely been considered in the context of mycotoxin exposure. The current review aimed at providing a summary of DON, ZEN, OTA, FB1, AFB1, and PAT effects on intestinal barrier function, with special focus on mucus and microbiota. DON, ZEN, OTA, FB1, AFB1, and PAT are known to markedly affect epithelial cell integrity and functions. Regarding mucus, DON is the most documentated mycotoxin. In vivo, toxicological impact of DON generally has only been assessed through goblet cell number. Evaluation of the mycotoxins/mucus interplay considering other indicators such as composition, thickness, and penetrability of mucus, mucin O-glycosylation thus warrants further attention. With respect to microbiota, few short-term studies to date have been reported indicating deleterious effects. However, long-term exposure to mycotoxins may also produce significant changes in microbiota composition and metabolic activity, which requires further experimentation. In conclusion, mucus and microbiota are key targets for dietary mycotoxins although assessment of induced effects is preliminary. A significant research effort is now underway to determine the adverse consequences of mycotoxins on mucus and microbiota considered as individual but also as tightly connected gut players.

Topics: Aflatoxin B1; Animals; Fumonisins; Gastrointestinal Microbiome; Humans; Intestinal Mucosa; Intestines; Mycotoxins; Ochratoxins; Patulin; Trichothecenes; Zearalenone

Mycotoxins are fungal secondary metabolites with bioaccumulation levels leading to their carry-over into animal fluids, organs, and tissues. As a consequence, mycotoxin determination in biological samples from humans and animals has been reported worldwide. Since most mycotoxins show toxic effects at low concentrations and considering the extremely low levels present in biological samples, the application of reliable detection methods is required. This review summarizes the information regarding the studies involving mycotoxin determination in biological samples over the last 10 years. Relevant data on extraction methodology, detection techniques, sample size, limits of detection, and quantitation are presented herein. Briefly, liquid-liquid extraction followed by LC-MS/MS determination was the most common technique. The most analyzed mycotoxin was ochratoxin A, followed by zearalenone and deoxynivalenol-including their metabolites, enniatins, fumonisins, aflatoxins, T-2 and HT-2 toxins. Moreover, the studies were classified by their purpose, mainly focused on the development of analytical methodologies, mycotoxin biomonitoring, and exposure assessment. The study of tissue distribution, bioaccumulation, carry-over, persistence and transference of mycotoxins, as well as, toxicokinetics and ADME (absorption, distribution, metabolism and excretion) were other proposed goals for biological sample analysis. Finally, an overview of risk assessment was discussed.

Topics: Aflatoxins; Animals; Chromatography, High Pressure Liquid; Chromatography, Liquid; Fumonisins; Humans; Mycotoxins; Ochratoxins; Spectrometry, Mass, Electrospray Ionization; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

Mycotoxins that commonly occur in cereal grains and other products can contaminate finished processed foods on account of their high toxicity. The mycotoxins that are commonly associated with food grains include aflatoxins, ochratoxin A, fumonisins, deoxynivalenol, and zearalenone. Various food-processing operations include sorting, trimming, cleaning, cooking, baking, frying, roasting, flaking, and extrusion that have variable effects on mycotoxins. The nature of the processing operation viz. physical, chemical, or thermal plays an important role in this; usually, the processes that utilize the higher temperatures have greater effects on mycotoxin dissipation. In general, the processes are known to reduce mycotoxin concentrations significantly, but do not eliminate them completely. However, roasting and extrusion processing result in lowest mycotoxin concentrations, since these involve higher temperatures. It is observed that very high temperatures are needed to bring about high reduction in mycotoxin concentrations, approaching acceptable background levels. The treatment with chemicals like ammonia, bicarbonate, citric acid, or sodium bisulfite is also effective in resulting in significant decline in mycotoxin concentrations.

Topics: Aflatoxins; Edible Grain; Food Contamination; Food Handling; Food Microbiology; Fumonisins; Hot Temperature; Mycotoxins; Ochratoxins; Trichothecenes; Zearalenone

This review summarizes the information regarding the in vivo studies of Fusarium mycotoxins in the last decade. The most common studies are classified as subacute toxicity, subchronic toxicity, acute toxicity, toxicokinetic studies and teratogenicity in order of importance. The most used animals in in vivo studies are pigs, rats, chickens and mice. Fumonisin B1, deoxynivalenol, zearalenone, nivalenol and T-2 toxin are the most studied fusarotoxins. Studies with combinations of mycotoxins are also frequent, deoxynivalenol generally being one of them. The predominant route of administration is oral, administered mostly in the form of naturally contaminated feed. Other administration routes also used are intraperitoneal, intravenous and subcutaneous. In vivo research on Fusarium mycotoxins has increased since 2010 highlighting the need for such studies in the field of food and feed safety.

Topics: Animal Feed; Animals; Chickens; Consumer Product Safety; Disease Models, Animal; Food Contamination; Food Microbiology; Fumonisins; Fusarium; Mice; Mycotoxins; Rats; Swine; T-2 Toxin; Trichothecenes; Zearalenone

The term mycotoxin was coined for toxic metabolites secreted by some fungi in food, food products and feed. The most prominent mycotoxins include aflatoxins (AFs), deoxynivalenol, zearalenone, ochratoxin, fumonisin and patulin. Among these some are proved to be strong carcinogenic agents such as AFs B1 while others are under suspicion to have carcinogenic effects. Ingestion of such mycotoxin-contaminated food and feed pose to a threat, mycotoxicoses. Various conventional techniques are available for the detection of mycotoxins, but unfortunately as a consequence of their constraint, the development of new and rapid techniques is the need of the hour. The use of nanotechnology for the development of nanobiosensors would be the alternative sensitive methods for the rapid detection of mycotoxins. Implementation of nanomaterials in the fabrication of nanobiosensors and their use for the detection of the mycotoxins in food and feed is the centre of interest of this review. We have inventoried nanomaterials applied for weaving nanobiosensors, which includes carbon nanotubes, nanowires, nanoparticles, quantum dots, nanorods and nanofibers. In addition, we have extensively reviewed available nanobiosensors specific for different mycotoxins, their advantages and challenges.

Topics: Aflatoxins; Animal Feed; Edible Grain; Food Contamination; Fumonisins; Humans; Mycotoxins; Nanostructures; Nanotechnology; Ochratoxins; Trichothecenes; Zearalenone

This review gives an overview of flow cytometry applications to toxicological studies of several physiological target sites of mycotoxins on different mammalian cell lines. Mycotoxins are secondary metabolites of fungi that may be present in food, feed, air and water. The increasing presence of mycotoxins in crops, their wide distribution in the food chain, and their potential for toxicity demonstrate the need for further knowledge. Flow cytometry has become a valuable tool in mycotoxin studies in recent years for the rapid analysis of single cells in a mixture. In toxicology, the power of these methods lies in the possibility of determining a wide range of cell parameters, providing valuable information to elucidate cell growth and viability, metabolic activity, mitochondrial membrane potential and membrane integrity mechanisms. There are studies using flow cytometry technique on Alternaria, Aspergillus, Fusarium and Penicillium mycotoxins including information about cell type, assay conditions and functional parameters. Most of the studies collected in the literature are on deoxynivalenol and zearalenone mycotoxins. Cell cycle analysis and apoptosis are the processes more widely investigated.

Topics: Alternaria; Animals; Aspergillus; Cell Line; Cell Survival; Flow Cytometry; Fusarium; Humans; Mammals; Membrane Potential, Mitochondrial; Mycotoxins; Penicillium; Trichothecenes; Zearalenone

Mycotoxins are secondary metabolites produced by fungi especially those belonging to the genus Aspergillus, Penicillum and Fusarium. Mycotoxin contamination can occur in all agricultural commodities in the field and/or during storage, if conditions are favourable to fungal growth. Regarding animal feed, five mycotoxins (aflatoxins, deoxynivalenol, zearalenone, fumonisins and ochratoxin A) are covered by EU legislation (regulation or recommendation). Transgressions of these limits are rarely observed in official monitoring programs. However, low level contamination by Fusarium toxins is very common (e.g., deoxynivalenol (DON) is typically found in more than 50% of the samples) and co-contamination is frequently observed. Multi-mycotoxin studies reported 75%-100% of the samples to contain more than one mycotoxin which could impact animal health at already low doses. Co-occurrence of mycotoxins is likely to arise for at least three different reasons (i) most fungi are able to simultaneously produce a number of mycotoxins, (ii) commodities can be contaminated by several fungi, and (iii) completed feed is made from various commodities. In the present paper, we reviewed the data published since 2004 concerning the contamination of animal feed with single or combinations of mycotoxins and highlighted the occurrence of these co-contaminations.

Topics: Aflatoxins; Animal Feed; Animals; Aspergillus; Europe; Food Contamination; Fumonisins; Fusarium; Guidelines as Topic; Mycotoxins; Ochratoxins; Trichothecenes; Zearalenone

The contamination of cereal grains with toxic secondary metabolites of fungi, mycotoxins, is a permanent challenge in animal nutrition as health and performance of the animals may be compromised as well as the quality of animal derived food. Therefore the present article reviews the issue of mycotoxins in animal nutrition. As the Fusarium toxins deoxynivalenol (DON) and zearalenone (ZON) are of particular importance under the production conditions in central Europe and Germany, with respect to their frequent occurrence in toxicologically relevant concentrations, special emphasis is layed on those mycotoxins. The effects of DON and ZON on susceptible animals as well as management strategies to cope with the contamination of grain with those toxins are reviewed.

Topics: Animal Feed; Animals; Food Contamination; Trichothecenes; Zearalenone

This review summarizes the toxicological data on the effects of the mycotoxins zearalenone (ZON), its metabolites, and deoxynivalenol (DON) on different parameters relating to reproductive and non-reproductive organs in female pigs. In vivo, 22 mg ZON kg(-1) in the diet cause alterations in the reproductive tract of swine such as in the uterus, and affects follicular and embryo development. ZON and its metabolites have been shown to bind competitively to oestrogen receptors in an in vitro system. The feeding of pigs with a 9 mg DON kg(-1)-contaminated diet can act on protein synthesis, humoral and cellular immune response depending on dose, exposure and timing of functional immune assay, and affect liver and spleen cell structures. Beside these effects, reproductive alterations were observed in pigs, too. Both in vivo and in vitro exposure to DON decreased oocyte and embryo development. In vitro application of DON to uterine cells inhibits their proliferation rate and modulates the process of translation at a different molecular level when compared with the in vivo application. The histopathological results provide evidence of spleen and liver dysfunction in the absence of clinical signs, especially in pigs fed higher concentrations of Fusarium toxin-contaminated wheat. Prepuberal gilts react more sensitively to DON > ZON feeding compared with pregnant sows. In the liver, histopathological changes such as glycogen decrease and interlobular collagen uptake were only observed in prepuberal gilts, whereas enhancement of haemosiderin was found in both perpuberal gilts and pregnant sows. This review presents some of the current knowledge on the biological activities of ZON and DON in pig. Altogether, ZON affects reproduction of pigs most seriously because it possesses oestrogenic activity. However, DON affects reproduction in pigs via indirect effects such as reduced feed intake, resulting in reduced growth or impairment of function in vital organs such as liver and spleen.

Topics: Animal Feed; Animals; Female; Food Contamination; Genitalia, Female; Liver; Mycotoxins; Spleen; Swine; Trichothecenes; Uterus; Zearalenone

Extrusion cooking is one of the fastest growing food-processing operations in recent years due to several advantages over traditional methods. Apart from its main goal of improving the quality of intermediate and final processed products, it may incidentally also improve safety because of the potential to reduce mycotoxin levels in cereals. This review is focused on extrusion cooking and aims to give a general overview of its impact in reducing mycotoxin levels in cereals. Extrusion cooking generally decreases the mycotoxins levels at rates depending on different factors such as the type of extruder, the type of screw, the die configuration, the initial mycotoxin concentration, the barrel temperature, the screw speed, the moisture content of the raw material and the use of additives. Reductions of 100, 95 and 83% for fumonisins, aflatoxins and zearalenone, respectively, have been reported during extrusion cooking of cereals, while lower reductions were observed for deoxynivalenol, ochratoxin A and moniliformin, where maximum reductions did not exceed 55, 40 and 30%, respectively.

Topics: Aflatoxins; Carcinogens; Carcinogens, Environmental; Cooking; Cyclobutanes; Edible Grain; Estrogens, Non-Steroidal; Food Contamination; Fumonisins; Hydrolysis; Mycotoxins; Ochratoxins; Trichothecenes; Zearalenone

The objectives of this review are to provide 1) information on the FDA Feed Contaminants Program, 2) the legal history of aflatoxins and their current action levels, 3) a report on the levels of aflatoxins, fumonisins, vomitoxin, ochratoxin A, and zearalenone in domestic and import surveillance samples of feed during fiscal years 1989 through 1992, and 4) information on naturally occurring toxins encountered recently by the Center for Veterinary Medicine. Ten of 644 (1.6%) domestic corn samples and 7 of 106 (6.6%) domestic cottonseed samples contained aflatoxins at levels > 300 ppb. The mean fumonisin level in the 1990 survey of 85 corn screening samples was 12.1 ppm, and the values ranged from 2.6 to 32 ppm. The mean vomitoxin levels in the 1991 survey of 207 winter wheat samples and 206 spring wheat samples was 2.4 and .9 ppm, respectively. Ochratoxin A was not detected in 168 samples. Zearalenone was detected at levels > .15 ppm in only 1 of 161 samples. Cottonseed containing 13,000 ppm gossypol was recently implicated in the deaths of dairy cows. Crambe meal and canola meal are sanctioned for use in feed with certain restrictions, including the levels of glucosinolates. The FDA is continuing its surveillance and will strive to provide guidance on the increasing number of naturally occurring toxins.

Topics: Aflatoxins; Alkaloids; Animal Feed; Animals; Animals, Domestic; Erucic Acids; Food Contamination; Fumonisins; Glucosinolates; Gossypol; Mycotoxins; Ochratoxins; Toxins, Biological; Trichothecenes; Zearalenone

Aflatoxins, zearalenone, deoxynivalenol, fumonisins, and their respective metabolites require specific procedures for their determination because of their diverse chemistry and occurrence in complex matrices of feedstuffs and foods. Major sources of error in the analysis of these mycotoxins arise from inadequate sampling and inefficient extraction and cleanup procedures. The determinative step in the assay for each of these toxins is sensitive to levels below those that are considered detrimental to humans and animals. Aflatoxins can be determined in grains and animal fluids and tissues by TLC, HPLC, gas chromatography-mass spectrometry (GC-MS), and ELISA procedures. Zearalenone, an estrogenic mycotoxin, can readily be determined in cereal grains and foods by HPLC (50 ng/g) and by TLC (300 ng/g). No incurred levels of zearalenone or its metabolites have been detected in animal tissues destined for human consumption. Deoxynivalenol can be determined in wheat and corn at 300 ng/g by a rapid TLC procedure and at 325 ng/g by a GC method. Although not tested collaboratively, an HPLC procedure and an ELISA screening procedure are capable of detecting deoxynivalenol at low (nanograms/gram) levels in feedstuffs and foods. The recently characterized fumonisins can be detected by TLC, HPLC, and GC-MS at levels below those now considered harmful. Thin-layer chromatography and HPLC (with fluorescence detection of derivatives) procedures can detect fumonisins at approximately 100 ng/g; GC-MS is required for detection at lower levels.

Topics: Aflatoxins; Animal Feed; Animals; Food Contamination; Food Microbiology; Humans; Mycotoxins; Trichothecenes; Zearalenone

Topics: Animal Feed; Animals; Carcinogens; Edible Grain; Food Analysis; Foodborne Diseases; Humans; Mycotoxins; Neoplasms; Trichothecenes; Zearalenone

The author reports a number of practical cases, indicating that moulds and mycotoxins may play an important role in Dutch pig farming. Therefore it seems desirable to him to form a study group that after investigation advises how problems with mycotoxins in farm animals can be restricted in the future.

Topics: Aflatoxins; Animals; Mycotoxins; Ochratoxins; Swine; Swine Diseases; Trichothecenes; Zearalenone

Mycotoxins are metabolic products of mycotoxins which have various chemical structures and show various toxic effects. Deoxynivalenol (vomitoxin) is an important economic factor in pig production due to growth depression and suppression of the immune system. Previous studies have shown that the 1-ppm limit in the sole feed for pigs should not be exceeded. Studies of methods of detoxification have as yet not produced conclusive results. Zearalenone has an tolerable effect and may lead to fertility disturbances on the oestrogen production in pigs and can cause remarkable economic damage even in the ppb range. Recommendations of upper limits cannot be made on the basis of the available results. The kidney toxin ochratoxin A is of importance in pig and poultry breeding and--due to its accumulation in the tissue--represents a possible source of danger to man. Since a possible carcinogenic effect of the toxin cannot be excluded, its content in animal rations should be kept as low as possible. For ruminants mycotoxins as a whole do not represent a particular source of danger as these substances can be degraded or converted by rumen flora.

Topics: Animal Feed; Animals; Animals, Domestic; Cattle; Drug Residues; Female; Food Contamination; Humans; Male; Ochratoxins; Poultry; Pregnancy; Swine; Trichothecenes; Zearalenone

Molds are parasitic plants that are ubiquitous in livestock feedstuffs. Even though molds themselves reduce the quality of grains, their synthesis of chemical substances termed mycotoxins causes the greatest monetary loss to the animal industry. Five major mycotoxins that impair growth and reproductive efficiency in North America are aflatoxins, zearalenone, deoxynivalenol, ochratoxin, and ergot. Aflatoxins are produced by Aspergillus flavus and Aspergillus parasiticus. Consumption of grains containing aflatoxins by swine affects reproduction indirectly by reducing feed intake and growth. In swine, aflatoxins impair liver and kidney function, delay blood clotting, increase susceptibility to bruising, and interfere with cellular humoral immune systems. Ruminants are comparatively resistant to aflatoxicosis, but presence of aflatoxins in milk of dairy cows is closely monitored for human safety. Depending on environmental conditions, Fusarium roseum can produce either zearalenone or deoxynivalenol. Days 7 to 10 postmating seem to be a critical period of gestation for zearalenone to exert its detrimental actions on early embryonic development. Presence of deoxynivalenol in swine feedstuffs decreases feed intake, causes feed refusal, and induces occasional vomiting. Several species of Penicillium and Aspergillus produce ochratoxin, a mycotoxin that causes necrosis of kidney tissue. Ergot alkaloids produced by Claviceps purpurea on wheat can cause reproductive problems and are associated with lactational failure in swine. Various methods have been developed to remove mycotoxins from infected feedstuffs. Chemical analyses in laboratories as well as diagnostic kits suitable for use at the elevator or farm can be used successfully to identify which mycotoxins are present in suspect feedstuffs.

Topics: Aflatoxins; Animals; Animals, Domestic; Aspergillus; Claviceps; Ergotism; Fusarium; Mycotoxicosis; Mycotoxins; Ochratoxins; Penicillium; Reproduction; Trichothecenes; Zearalenone

Topics: Animal Feed; Animals; Cyclobutanes; Food Contamination; Food Microbiology; Fusarium; Humans; Mutagens; Mycotoxins; Polyenes; Trichothecenes; Zearalenone

Sows were fed naturally contaminated diets containing: (i) 100 ppb zearalenone (ZEN) one week before farrowing and during the lactation period (at 26 days), (ii) 100 ppb ZEN one week before farrowing and 300 ppb ZEN during the lactation period, or (iii) 300 ppb ZEN one week before farrowing and during the lactation period. All diets contained 250 ppb deoxynivalenol (DON). The highest levels of ZEN, α-ZEL, or β-ZEL were observed in the serum of sows fed 300 ppb ZEN before farrowing and during lactation. However, only α-ZEL was significantly increased in the colostrum and milk of these sows. Sows fed the 300 ppb ZEN during the complete trial presented a significant decrease in backfat thickness before farrowing. This effect was accompanied by a decrease in serum leptin levels. These sows also presented a decrease in estradiol levels and this effect was observed in their piglets exposed during lactation, which presented increased glucagon-like peptide 1, but no changes in serum levels of ZEN, α-ZEL, or β-ZEL. Although all sows were fed the same levels of DON, the serum levels of DON and de-epoxy-DON were increased only in the serum of piglets from the sows fed a diet with the highest ZEN levels during the whole experimental period. Moreover, these piglets presented gut inflammation, as indicated by significantly increased calprotectin levels in their serum.

Topics: Animal Feed; Animals; Diet; Drug Residues; Female; Food Contamination; Lactation; Milk; Swine; Trichothecenes; Zearalenone

Topics: Animal Feed; Animals; Bilirubin; Creatinine; European Union; Fatty Acids; Food Contamination; Fumonisins; Fusarium; gamma-Glutamyltransferase; Glutathione; Kidney; Lipid Metabolism; Liver; Phospholipids; Spleen; Swine; Trichothecenes; Urea; Weaning; Zearalenone

To investigate the usefulness of follicular fluid (FF) in relation to blood plasma and bile as indicators of exposure of dairy cows to ZEN, DON and their metabolites, a dose-response study was performed with 30 dairy cows. The cows, 10 in each group (named CON; FUS-50, FUS-100), received a diet with three different concentrations of Fusarium toxin-contaminated maize. Thereby, the following dietary concentration were reached: CON (0.02 mg ZEN and 0.07 mg DON, per kg dry matter, DM), FUS-50 (0.33 mg ZEN and 2.62 mg DON, per kg DM) and FUS-100 (0.66 mg ZEN and 5.24 mg DON, per kg DM). ZEN, DON and de-epoxy-DON (de-DON) were detected in FF. Based on the linear regression between toxin concentration in plasma and FF, it seems that about 50% (m = 0.5) of ZEN present in plasma is present in FF while an increase of 1 ng/ml DON or de-DON in plasma is paralleled by an increase of 1.5 ng/ml DON or 1.1 ng/ml de-DON in FF. ZEN, DON and their metabolites, except zearalenone (ZAN), were also detected in bile. Contrary to DON and de-DON, ZEN and its metabolites were accumulated in bile so that the concentration of ZEN and metabolites was much higher than for DON and de-DON. The main compound was β-zearalenol (β-ZEL). The biliary ZEN, α-zearalenol (α-ZEL) and β-ZEL concentration correlated linearly with each other with an uncertainty of <15% (r(2) ≥ 0.86), whereas the ratio between ZEN: α-ZEL: β-ZEL was about 1.5:1:11. With the help of established linear relationship between toxin intake and toxin concentration, bile could be used as diagnostic indicator to assess the exposure of cows.

Topics: Animals; Bile; Cattle; Cattle Diseases; Drug Residues; Female; Follicular Fluid; Trichothecenes; Zearalenone

This study was conducted to investigate the effects of deoxynivalenol (DON) and zearalenone (ZEA) on some biochemical indices of broiler chickens. Twenty-four Ross 308 hybrid broiler chickens of both sexes were fed diets containing maize contaminated with Fusarium mycotoxins. The diets included a control diet (DON 0.60 mg/kg feed; ZEA 0.07 mg/kg feed), an experimental 1 diet (DON 3.4 mg kg⁻¹ feed; ZEA 3.4 mg kg⁻¹ feed), and an experimental 2 diet (DON 8.2 mg kg⁻¹ feed; ZEA 8.3 mg kg⁻¹ feed). Contaminated diets were fed from 14 days of age for 14 days. Blood samples were collected from 4-week-old birds. Chicks fed a diet containing a low level of contaminated maize (experimental 1) had decreased plasma potassium, magnesium, phosphorus, total protein, albumin, triglycerides, free glycerol concentrations and increased cholesterol and calcium levels as well as alkaline phosphatase (ALP) and aspartate aminotransferase (AST) enzyme activities as compared to the control. Feeding a diet contaminated with high levels of mycotoxins (experimental 2) resulted in decreased plasma potassium, magnesium, total protein, albumin, triglycerides, free glycerol concentrations and increased plasma ALP, alanine aminotransferase (ALT) and AST enzyme activities. The effect of mycotoxin-contaminated diets on ALP activity was dose dependent. Chloride concentration was not affected by the diets. It can be concluded that feeding diets contaminated with both levels of Fusarium mycotoxins significantly affected protein, lipid and mineral metabolism as well as AST and ALP enzyme activities in broiler chickens.

Topics: Alanine Transaminase; Albumins; Alkaline Phosphatase; Animal Feed; Animals; Blood Proteins; Chickens; Cholesterol; Diet; Female; Food Contamination; Fusarium; Glycerol; Male; Potassium; Trichothecenes; Triglycerides; Zearalenone

Fusarium spp. moulds are common in moderate climate regions of North America, Asia and Europe. They produce hepatotoxic and nephrotoxic mycotoxins, acting like estrogens, impairing hemopoesis and immunosuppressing. Actively dividing skin cells, lymphatic tissue, haemopoetic tissue and gastrointestinal tissue are the most sensitive for these trichothecenes action. The mucosal membrane of the gastrointestinal tract is the first barrier of the organism contacting with foreign antigens like feed proteins, natural toxins, saprophytic and pathogenic microflora and mycotoxins. The aim of this study was to perform histological estimation of the porcine small intestine after short term intoxication with low doses of deoxynivalenol (DON), T-2 toxin (T-2) and zearalenone (ZEA) obtained from wheat naturally contaminated with Fusarium moulds. Experimental pigs (n=5) were fed for 14 days feed containing DON, T-2 and ZEA (28.9, 11.5 and 33.2 microg kg(-1) of feed). On the last day of the experiment, the animals were euthanised and samples of the jejunum were collected for histological examination. In the experimental pigs, normally developed intestinal villi and crypts were found. However, number of acidophilic granulocytes in the mucous membrane and decreased numbers of goblet cells, increased numbers of endothelial lymphocytes and numerous plasma cells in intestinal epithelium was observed. On the surface of the intestinal epithelium the glycocalyx was poorly developed. The results obtained suggest that short term intoxication with low doses of DON, T-2 and ZEA does not cause significant changes in the histological structure of the small intestine in the pig. However, low concentrations of DON, T-2 and ZEA probably influence enterocytes metabolism and evoke inflammation of the mucous membrane of the small intestine.

Topics: Animal Feed; Animals; Food Contamination; Intestinal Diseases; Intestinal Mucosa; Intestine, Small; Swine; Swine Diseases; T-2 Toxin; Trichothecenes; Zearalenone

This study was to investigate the effects of the combination of deoxynivalenol (DON) and zearalenone (ZON) on pigs. Twenty-four weaning piglets were divided into a control group fed a diet free of mycotoxins and a toxin group fed a diet containing 1 mg/kg DON and 250 microg/kg ZON. The results showed that supplementation of DON and ZON in diets had extensive effects on pigs. More specifically, DON and ZON caused levels of total protein, albumin, and globulin in sera to decrease (p < 0.05) by 14.5%, 6.5% and 11.3%, respectively, and at the same time increased (p < 0.05) the serum enzyme activities of gamma-glutamyltransferase, aspartate aminotransferase and alanine aminotransferase by 72.0%, 32.6% and 36.6%, respectively. In addition, DON and ZON decreased (p < 0.05) the level of anticlassical swine fever antibody titers by 14.8%. Real-time PCR showed that DON and ZON caused the mRNA expression levels of IFN-gamma, TNF-alpha, IL-2, to decrease (p < 0.05) by 36.0%, 29.0% and 35.4%, respectively. Histopathological studies demonstrated that DON and ZON caused abnormalities in the liver, spleen, lymph nodes, uterus, and kidney. The concentrations of DON and ZON used in this study are in line with the published critical values permitted by BML. Our study clearly put the standard and adequacy of safety measures for these toxins into question. The authors suggest that with the increasing availability of cellular and molecular technologies, it is time to revisit the safety standards for toxins in feeds so as to make feeds safer, providing consumers with safer products.

Topics: Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Diet; Drug Therapy, Combination; Female; Swine; Swine Diseases; Trichothecenes; Zearalenone

Topics: Chromatography, High Pressure Liquid; Edible Grain; Food Contamination; Mycotoxins; Tandem Mass Spectrometry; Zearalenone

Topics: Animals; Brazil; Edible Grain; Food Contamination; Fusarium; Hordeum; Humans; Mycotoxins; Zearalenone

The study aimed to investigate toxicity and the mechanism of toxicity of two

Topics: Cell Cycle; Cell Proliferation; DNA; Hep G2 Cells; Humans; Mycotoxins; Zearalenone

The common mycotoxins in polluted grains are aflatoxin B1(AFB1), zearalenone (ZEN) and deoxynivalenol (DON). Because of the potential threat to humans and animals, it is necessary to detect mycotoxin contaminants rapidly. At present, later flow immunoassay (LFIA) is one of the most frequently used methods for rapid analysis. However, multistep sample pretreatment processes and organic solvents are also required to extract mycotoxins from grains. In this study, we developed a one-step and "green" sample pretreatment method without using organic solvents. By combining with LFIA test strips and a handheld detection device, an on-site method for the rapid detection of AFB1, ZEN and DON was developed. The LODs for AFB1, ZEN and DON in corn are 0.90 μg/kg, 7.11 μg/kg and 10.6 μg/kg, respectively, and the working ranges are from 1.25 μg/kg to 40 μg/kg, 20 μg/kg to 2000 μg/kg and 35 μg/kg to 1500 μg/kg, respectively. This method has been successfully applied to the detection of AFB1, ZEN and DON in corn, rice and peanut, with recoveries of 89 ± 3%-106 ± 3%, 86 ± 2%-108 ± 7% and 90 ± 2%-106 ± 10%, respectively. The detection results for the AFB1, ZEN and DON residues in certified reference materials by this method were in good agreement with their certificate values.

Topics: Aflatoxin B1; Animals; Arachis; Food Contamination; Humans; Mycotoxins; Oryza; Zea mays; Zearalenone

Topics: Avena; Chromatography, Liquid; Edible Grain; Food Contamination; Fusarium; Glucosides; Mycotoxins; Scotland; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes, Type B; Zearalenone

Edible insects are a solid alternative to meet the growing demand for animal protein. However, there are doubts regarding the safety of insect consumption. Mycotoxins are substances of concern for food safety, as they may cause harmful effects on the human organism and accumulate in the tissues of some animals. This study focuses on the characteristics of the main mycotoxins, the mitigation of human consumption of contaminated insects, and the effects of mycotoxins on insect metabolism. To date, studies reported the interaction of the mycotoxins aflatoxin B1, ochratoxin A, zearalenone, deoxynivalenol, fumonisin B1, and T-2, isolated or combined, in three insect species from Coleoptera and one from Diptera order. The use of rearing substrates with low mycotoxin contamination did not reduce the survival and development of insects. Fasting practices and replacing contaminated substrate with a decontaminated one decreased the concentration of mycotoxins in insects. There is no evidence that mycotoxins accumulate in the tissues of the insects' larvae. Coleoptera species showed high excretion capacity, while Hermetia illucens had a lower excretion capacity of ochratoxin A, zearalenone, and deoxynivalenol. Thus, a substrate with low mycotoxin contamination could be used for raising edible insects, particularly from the Coleoptera order.

Topics: Animals; Coleoptera; Diptera; Edible Grain; Edible Insects; Food Contamination; Humans; Mycotoxins; Zearalenone

Female pigs respond sensitive both to DON and ZEN with anorexia and endocrine disruption, respectively, when critical diet concentrations are exceeded. Therefore, the frequent co-contamination of feed by DON and ZEN requires their parallel inactivation. The additive ZenA hydrolyzes ZEN while SBS inactivates DON through sulfonation. Both supplements were simultaneously added (+, 2.5 g SBS and 100 U ZenA/kg) to a control diet (CON-, 0.04 mg DON and < 0.004 mg ZEN/kg; CON+, 0.03 mg DON and < 0.004 mg ZEN/kg) and a Fusarium toxin contaminated diet (FUS-, 2.57 mg DON and 0.24 mg ZEN/kg; FUS+, 2.04 mg DON and 0.24 mg ZEN/kg). The 4 diets were fed to 20 female weaned piglets each (6 kg initial body weight) for 35 days; the piglets were sacrificed thereafter for collecting samples. Supplements improved performance and modified metabolism and hematology independent of dietary DON contamination. The mechanisms behind these changes could not be clarified and require further consideration. SBS reduced DON concentration in feed by approximately 20% and to the same extent in blood plasma and urine suggesting that no further DON sulfonate formation occurred in the digestive tract before absorbing DON in the upper digestive tract or that additionally formed DON sulfonates escaped absorption. DON sulfonates were detected in feces suggesting that unabsorbed DON sulfonates reached feces and/or that unabsorbed DON was sulfonated in the hindgut. The observed reduction rate of 20% was evaluated to be insufficient for feeding practice. Galenic form of SBS added to dry feed needs to be improved to support the DON sulfonation in the proximal digestive tract.ZenA was active in the digestive tract as demonstrated by the presence of its hydrolyzed none-estrogenic reaction products hydrolyzed ZEN (HZEN) and decarboxylated and hydrolyzed ZEN (DHZEN) both in feces, systemic circulation, and urine of group FUS+ compared to group FUS-. The presence of these hydrolysis products was paralleled by a significant decrease in high-estrogenic ZEN concentrations which, in turn, was related to a decrease in relative weights of uteri and ovaries when compared to group FUS-. Thus, ZenA was proven to be effective; both in terms of biomarkers and biological effects.

Topics: Animal Feed; Animals; Female; Food Contamination; Fusarium; Hydrolases; Swine; Trichothecenes; Zearalenone

There is a possibility for exposed lactating mammalians to transfer some contaminants to their milk. This study aimed to determine the levels and changes of Zearalenone (ZEN), Deoxynivalenol (DON) mycotoxins for the first five months in human milk.. Voluntary lactating mothers having infants with gestational length ≥ 37 weeks were enrolled between August 2017 and June 2018 in Şanlıurfa. Mothers and infants with chronic health problems were not included in the study. Human milk samples were taken at three different times; on enrollment (Day 6-10, visit 1), between 4 and 6 weeks postpartum (visit 2), and between 14 and 19 weeks postpartum (visit 3). Mycotoxin levels in human milk were measured utilizing Helica brand commercial kit.. Nineteen voluntary mothers and their breastfed infants with three human milk samples completed the study. The mean ages of mothers and infant (± SD) were 27.4 (± 5.4) years and 7.6 (± 0.9) days on enrollment. Median levels of ZEN and DON in human milk samples were 0.39 and 16.7 ng/mL, respectively. None of the cases had a ZEN daily intake higher than 250 ng/kg bw per day. However, three fourth of the cases had DON intake higher than > 1000 ng/kg bw per day. When adjusted for infant weight for age and sex, both ZEN levels and daily intake were decreased progressively from visit 1 to visit 3 (p < 0.001). DON levels and daily intake at visit 2 were found to be significantly lower in samples of visit 3 than that taken in visit 2 (p = 0.004 and p < 0.001, respectively).. Breast milk monitoring study revealed that ZEN and DON mycotoxins were present in the mother-infant environment. Contamination levels changed during the lactation period.

Topics: Adult; Animals; Female; Follow-Up Studies; Humans; Infant; Lactation; Mammals; Milk, Human; Mycotoxins; Young Adult; Zearalenone

Deoxynivalenol (DON) and zearalenone (ZEN) are often detected in plant materials used to produce feed for pre-pubertal gilts. Daily exposure to small amounts of these mycotoxins causes subclinical conditions in pigs and affects various biological processes (e.g. mycotoxin biotransformation). The aim of this preclinical study was to evaluate the effect of low monotonic doses of DON and ZEN (12 µg/kg body weight-BW-and 40 µg/kg BW, respectively), administered alone or in combination to 36 prepubertal gilts for 42 days, on the degree of immunohistochemical expression of oestrogen receptors (ERs) in the liver and the mRNA expression of genes encoding selected liver enzymes during biotransformation processes. The level of expression of the analysed genes proves that the tested mycotoxins exhibit variable biological activity at different stages of biotransformation. The biological activity of low doses of mycotoxins determines their metabolic activity. Therefore, taking into account the impact of low doses of mycotoxins on energy-intensive processes and their endogenous metabolism, it seems that the observed situation may lead to the activation of adaptation mechanisms.

Topics: Animals; Female; Liver; Mycotoxins; Receptors, Estrogen; Sus scrofa; Swine; Zearalenone

Cattle are deemed less susceptible to mycotoxins due to the limited internal exposure resulting from rumen microbiota activity. However, the significant amounts of Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) frequently detected in bovine follicular fluid samples suggest that they could affect ovarian function. Both mycotoxins trigger several patterns of cell death and activate the NLRP3 inflammasome in the intestine. In vitro studies have reported a number of adverse effects on bovine oocytes. However, the biological relevance of such findings with regard to realistic concentrations of DON and ZEN in bovine follicular fluid is still not clear. Hence, it is important to better characterize the effects of dietary exposure to DON and ZEN on the bovine ovary. Using bovine primary theca cells, this study investigated the effects of real-life patterns for bovine ovary exposure to DON and ZEN, but also DON metabolite DOM-1, on cell death and NLRP3 inflammasome activation. Exposure to DON starting from 0.1 μM significantly decreased theca cell viability. The kinetics of phosphatidylserine translocation and loss of membrane integrity showed that ZEN and DON, but not DOM-1, induce an apoptotic phenotype. qPCR analysis of the expression of NLRP3, PYCARD, IL-1β, IL-18, and GSDMD in primary theca cells at concentrations of mycotoxin previously reported in cow follicular fluid clearly indicated that DON and DOM-1 individually and in mixture, but not ZEN, activate NLRP3 inflammasome. Altogether, these results suggest that real-life dietary exposure of cattle to DON may induce inflammatory disorders in the ovary.

Topics: Animals; Apoptosis; Cattle; Female; Fusarium; Inflammasomes; Mycotoxins; NLR Family, Pyrin Domain-Containing 3 Protein; Theca Cells; Zearalenone

Fusarium species are common fungal pathogens of maize. Fusarium graminearum and Fusarium verticillioides, among others, can cause maize ear rot, and they are also mycotoxin producers. The aims of this work were to determine the frequency and diversity of Fusarium species in Uruguayan maize kernels, evaluate the toxigenic potential of the isolates, determine toxin contamination levels on freshly harvested grain, and assess the sensitivity of main Fusarium species against fungicides. Fusarium verticillioides was the most frequent species isolated, followed by Fusarium graminearum sensu stricto. Of F. verticillioides isolates studied for fumonisin production, 72% produced fumonisin B1 and 32% fumonisin B2. Considering in vitro toxin production by F. graminearum sensu stricto isolates, deoxynivalenol was the main toxin produced, followed by zearalenone and nivalenol. Fumonisins were the most frequently found toxins on freshly harvested maize samples (98% in 2018 and 86% in 2019), and also, fumonisin B1 was the toxin with highest concentration in both years studied (4860 µg/kg in 2018 and 1453 µg/kg in 2019). Deoxynivalenol and zearalenone were also found as contaminants. Metconazole and epoxiconazole were the most effective fungicides tested on F. verticillioides isolates. Fusarium graminearum sensu stricto isolates also were more sensitive to metconazole compared to other fungicides; nevertheless, epoxiconazole was less efficient in controlling this species. This is the first study that reports Fusarium species and mycotoxin contamination levels associated with maize grain in Uruguay. Its detection is the main step to develop management strategies in order to minimize fungal infection in maize crops.

Topics: Edible Grain; Food Contamination; Fumonisins; Fungicides, Industrial; Fusarium; Mycotoxins; Uruguay; Zea mays; Zearalenone

Mycotoxins are secondary fungal metabolites which pose a significant threat for global food and feed security [...].

Topics: Animal Feed; Animals; Food Contamination; Mycotoxins; Trichothecenes; Zearalenone

Zearalenone (ZEA) and deoxynivalenol (DON) are widely found in various feeds, which harms livestock's reproductive health. Both mitochondria and endoplasmic reticulum (ER) can regulate cell apoptosis. This study aimed to explore the regulatory mechanism of endoplasmic reticulum stress (ERS) on ZEA- combined with DON-induced mitochondrial pathway apoptosis in piglet Sertoli cells (SCs). The results showed that ZEA + DON damaged the ultrastructure of the cells, induced apoptosis, decreased mitochondrial membrane potential, promoted the expression of cytochrome c (CytC), and decreased the cell survival rate. Furthermore, ZEA + DON increased the relative mRNA and protein expression of

Topics: Animals; Apoptosis; Caspase 3; Endoplasmic Reticulum Stress; Male; Mitochondria; Proto-Oncogene Proteins c-bcl-2; RNA, Messenger; Sertoli Cells; Swine; Tumor Suppressor Protein p53; Zearalenone

Deoxynivalenol (DON) and zearalenone (ZEA), which are commonly found in feed products, exhibit serious negative effects on the reproductive systems of domestic animals. However, the toxicity of mycotoxins on the uterine function of donkey (Equus asinus) remains unclear. This study investigated the biological effects of DON and ZEA exposure on donkey endometrial epithelial cells (EECs). It was administered 10 μM and 30 μM DON and ZEA to cells cultured in vitro. The results showed that 10 μM DON exposure markedly changed the expression levels of pyroptosis-associated genes and that 30 μM ZEA exposure changed the expression levels of inflammation-associated genes in EECs. The mRNA expression of cancer-promoting genes was markedly upregulated in cells exposed to DON and 30 μM ZEA; in particular, 10 μM and 30 μM DON and ZEA markedly disturbed the expression of androgen and estrogen secretion-related genes. Furthermore, Q-PCR, Western blot, and immunofluorescence analyses verified the different expression patterns of related genes in DON- and ZEA-exposed EECs. Collectively, these results illustrated the impact of exposure to different toxins and concrete toxicity on the mRNA expression of EECs from donkey in vitro.

Topics: Animals; Epithelial Cells; Equidae; Mycotoxins; Trichothecenes; Zearalenone

Raw Ca-based montmorillonite (MMT) was treated by H

Topics: Acids; Adsorption; Aflatoxin B1; Bentonite; Calcium; Inactivation, Metabolic; Mycotoxins; Temperature; Trichothecenes; Zearalenone

Mycotoxins can impart different types of combined toxicity to humans and animals, therefore, it is critical to understand the underlying mechanisms to eliminate the harm. Herein a combination of zearalenone (ZEA) at 2 μM and deoxynivalenol (DON) at 0.1 μM decreased cell viability and increased ROS level in HepG2 cells, suggesting synergistic toxicity exerted by ZEA and DON even at their low toxic concentrations. Moreover, apoptosis and inflammatory response were promoted after the co-exposure of ZEA and DON, indicated by the increased expression of BAX, Caspase-3, IL-1β and IL-6 genes. Such synergistic toxicity was closely associated with miR-221-mediated PTEN/PI3K/AKT signal pathway, with a negative regulatory relationship between PTEN and PI3K/AKT signaling. MiR-221 could influence cell viability and ROS level to counter the combined toxicity of ZEA and DON through targeting directly PTEN gene. This study demonstrated the toxicological impact of mycotoxin interactions on cells, and critical role of the interplay between miRNAs and PTEN in monitoring the synergistic toxicity of mycotoxin mixture.

Topics: Blotting, Western; Drug Combinations; Drug Synergism; Hep G2 Cells; Humans; Inhibitory Concentration 50; MicroRNAs; Oncogene Protein v-akt; Phosphatidylinositol 3-Kinase; PTEN Phosphohydrolase; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Trichothecenes; Zearalenone

Mycotoxins such as zearalenone (ZEN), deoxynivalenol (DON) and T-2 toxin (T-2) are the most poisonous biological toxins in food pollution. Mycotoxin contaminations are a global health issue. The aim of the current study was to use porcine Leydig cells as a model to explore the toxic effects and underlying mechanisms of ZEN, DON and T-2. The 50% inhibitory concentration (IC

Topics: 3-Hydroxysteroid Dehydrogenases; Animals; Apoptosis; Cholesterol Side-Chain Cleavage Enzyme; Leydig Cells; Male; Phosphoproteins; Progesterone; Swine; T-2 Toxin; Testosterone; Trichothecenes; Zearalenone

Regulations limiting aflatoxin levels in animal feed and guidance values for maximum levels for fumonisins (FB1 and FB2), deoxynivalenol (DON), ochratoxin A (OTA), zearalenone (ZON), HT-2, and T-2 toxins are in place both to protect animal health and to minimize potential transfer to animal products for human consumption. A multi-mycotoxin method which can handle complex feed matrices such as distillers dried grains with solubles (DDGS) is essential for analysis and accurate quantification without the need to revert to separately analyze individual mycotoxins.. The objective of this study is to generate single laboratory validation data for a method employing a multi-antibody immunoaffinity column (IAC) capable of providing cleanup for eleven mycotoxins, followed by LC-MS/MS quantification without the need for isotopic labelled and matrix-matched standards. The applicability of method is to be demonstrated for corn feed, pig feed, and DDGS by fortification and naturally occurring mycotoxins covering the range of regulated limits.. Feed sample (1 kg) ground by milling to approximately 1-2 mm particle size and sub-sample (5 g) extracted with acetonitrile-water-formic acid, passing through a multi-mycotoxin IAC, washing, and eluting prior to LC-MS/MS analysis monitoring selected ion transitions.. Recoveries were in the range 74 to 117% (excluding five outliers) for aflatoxins, FB1, FB2, DON, OTA, ZON, HT-2, and T2- toxins spiked into three commercial animal feed matrixes (n = 84) and within-day RSDs averaged 1.7 to 10.3% (n = 99).. Single laboratory validation of a multi-antibody IAC method coupled with LC-MS/MS has shown the method to be suitable for accurate quantification of eleven regulated mycotoxins in DDGS, pig feed, and poultry feed.. IAC method capable of accurately quantifying eleven regulated mycotoxins in complex feed matrices.

Topics: Aflatoxins; Animal Feed; Animals; Chromatography, Liquid; Food Contamination; Fumonisins; Humans; Mycotoxins; Ochratoxins; Swine; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

The main aim of this study was to determine the content of mycotoxins, such as: deoxynivalenol (DON), zearalenone (ZEA) and fumonisins (FUM) in cereal products, and such products intended for infants. The secondary objective was to assess consumer exposure to the DON, ZEA and FUM occurring in cereal products, including those intended for infants and young children.. The study included cereals and cereal products such as flours, grits, pastas, products of the bakery industry, snacks and cereal products intended for infants and young children, available in retail outlets in the Małopolska Province of Poland. DON content was determined by high-performance liquid chromatography with a DAD detector, while the contents of ZEA and FUM were detected by high-performance liquid chromatography with fluorescence detection.. The determined concentration of mycotoxins exceeded the maximum level specified in food law in only two cases. DON level in maize flour was 1511.0 μg kg. Levels higher than those permissible for the examined cereal products were noted in only two cases. FUMs were the most commonly found Fusarium mycotoxins, followed by DON and ZEA. The mean exposure doses of the assessed mycotoxins, resulting from the consumption of cereal products in the selected populations, were at low levels (reaching a maximum of 6.81%) and did not exceed the tolerable daily intake (TDI) or provisional maximum tolerable daily intake (PMTDI). Therefore, the observed average chronic exposure dose not pose a health risk to consumers.

Topics: Child; Child, Preschool; Edible Grain; Food Contamination; Fumonisins; Humans; Infant; Trichothecenes; Zearalenone

Deoxynivalenol (DON), which is one of the prevalent mycotoxins in food and feeds, exerts adverse effects on animal and human health. These effects are mainly associated with its ribotoxic properties, although few studies suggest the involvement of other mechanisms of action. To assess the ability of DON to disrupt estrogen signaling, we conducted an in vitro study using MCF-7 and MDA-MB-231 cells. After 72h, DON reduced cell viability in both cell lines, thus highlighting its well-known cytotoxic effect. However, after 6h, DON increased the expression of estrogen-responsive genes, hence demonstrating the stimulation of estrogen signaling by this mycotoxin after a short-term exposure. This effect was partially reversed by siRNA-mediated silencing of ERα expression and by 4-hydroxytamoxifen (ERα antagonist), but neither by G36 (GPER antagonist) nor by the siRNA-mediated silencing of PPARγ2 expression. Moreover, DON exposure induced an increase in the level of ERα phosphorylation at serine 167. Furthermore, when combined with zearalenone (a naturally co-occurring mycotoxin recognized as an endocrine disruptor), DON increased the expression of estrogen-responsive genes to a greater extent than each individual compound taken separately. Taken together, our results suggest, for the first time, that DON can disrupt estrogen signaling through the ligand-independent activation of ERα.

Topics: Animals; Estrogen Receptor alpha; Estrogens; Ligands; Mycotoxins; RNA, Small Interfering; Transcriptional Activation; Trichothecenes; Zearalenone

This study applied multi-mycotoxin liquid chromatography with tandem mass spectrometric detection (LC-MS/MS) methods to determine the biomarkers of exposure in urine and serum samples from a dose-response study with pigs. The 24 studied pigs were divided into three groups: a control and two experimental ones (with different levels of feed contamination). They were exposed to feed prepared from cereals contaminated with deoxynivalenol (DON), zearalenone (ZEN), ochratoxin A (OTA) and citrinin (CIT) for 14 days. After that, both experimental groups received the same feed as the control group for the next 14 days to determine the kinetics of the disappearance of mycotoxin biomarkers. Urine samples were collected daily in the morning and blood samples-eight-times during the experiment. The study reported herein was the first prolonged exposure experiment for multiple mycotoxins like OTA and CIT in pigs. The urinary and serum levels of all biomarkers correlated well with the respective toxin intake; thereby demonstrating that they are suitable biomarkers of exposure in pigs. Urine is a good candidate to monitor DON, ZEN, OTA, CIT exposure while serum may be used to monitor DON, OTA and CIT. Additionally, OTA has even been quantified in both matrices in the experimental groups two weeks after changing the contaminated feed back to the control, this result differed from those produced by the other mycotoxins which were only quantified during the first two weeks. Therefore both matrices are suitable candidates to monitor prolonged OTA exposure in pigs.

Topics: Animal Feed; Animals; Biomarkers; Citrinin; Female; Food Contamination; Ochratoxins; Sus scrofa; Trichothecenes; Zearalenone

Deoxynivalenol (DON) is a mycotoxin produced by fungi of. A total of 19 genital tracts (vagina, cervix, uterus, oviducts and ovaries) including the urinary bladder (n = 15) from reproductively failed gilts and different parity sows submitted from 8 farms in 2019/20 were examined pathomorphologically. DON as well as zearalenone (ZEA) were determined by using high performance liquid chromatography in 11 individual and 2 pooled (2 and 4 animals, respectively) bile samples. Microbiologic examinations of uterine (n = 17) and bladder (n = 12) specimens were additionally performed.. The results of the present study allow for the postulation that DON may cause fertility problems. Subsequent pathomorphological examinations of genital organs and the urinary bladder are recommended. When ≥ 2 organs are chronically inflamed and the uteri are additionally microbiologically positive, a contribution of DON may be assumed and confirmation via bile analysis is warranted.. Deoxynivalenol (DON) ist ein Mykotoxin, das von Pilzen der. Es wurden 19 Genitaltrakte (Vagina, Zervix, Uterus, Eileiter und Ovarien) inklusive Harnblasen (n = 15) fruchtbarkeitsgestörter Jung- und Altsauen unterschiedlicher Wurfnummern, die in den Jahren 2019/20 aus 8 Betrieben eingesandt wurden, pathomorphologisch untersucht. In 11 individuellen Galle- und 2 Poolproben (2 bzw. 4 Tiere) erfolgte die Bestimmung von DON und Zearalenon (ZEA) mittels Hochleistungsflüssigkeitchromatografie. Zudem unterlagen 17 Uteri und 12 Harnblasen einer mikrobiologischen Untersuchung.. Es wird postuliert, dass DON Fruchtbarkeitsprobleme verursachen kann. Zur Abklärung empfehlen sich pathomorphologische Untersuchungen der Genitalorgane und Harnblase. Sind ≥ 2 Organe chronisch entzündlich verändert und besteht zudem eine bakterielle Besiedelung des Uterus, ist eine Beteiligung von DON anzunehmen und durch eine Untersuchung im Gallensaft zu bestätigen.

Topics: Animals; Escherichia coli; Female; Mycotoxins; Pregnancy; Swine; Trichothecenes; Zearalenone

This study investigated the impact of malting of six wheat cultivars inoculated with

Topics: Biotransformation; Food Contamination; Food Handling; Food Microbiology; Fusarium; Trichothecenes; Triticum; Zearalenone

Mycotoxicoses in animals are caused by exposure to mycotoxin-contaminated feeds. Disease risk is managed using dietary adsorbing agents which reduce oral bioavailability. The objective of this work was to evaluate the efficacy of three selected yeast products as mycotoxin binders using in vitro and in vivo models. Their capacity to adsorb deoxynivalenol (DON), zearalenone (ZEA), and ochratoxin A (OTA) was evaluated using an in vitro model designed to simulate the pH conditions during gastric passage in a monogastric animal. Results showed that only one product, an enzymatic yeast hydrolysate (YHY) of a novel strain

Topics: Adsorption; Fungicides, Industrial; Mycotoxins; Ochratoxins; Poisons; Saccharomyces cerevisiae; Trichothecenes; Yeast, Dried; Zearalenone

A novel dual near-infrared fluorescence-based lateral flow immunosensor was developed to determine zearalenone and deoxynivalenol in maize. Two near-infrared dyes with distinct fluorescence characteristics were utilized to separately label the anti-zearalenone and anti-deoxynivalenol antibodies as detection reagents. The capture antigens zearalenone-BSA and deoxynivalenol-BSA were mixed and immobilized on the same test line of nitrocellulose membrane. This assay format facilitates simultaneous detection of the two mycotoxins on a single test line. After optimizing experimental parameters, the limits of detection for zearalenone and deoxynivalenol were as low as 0.55 μg/kg and 3.8 μg/kg in maize, respectively. The spiking experiment yielded recovery ratios ranging from 81.7% to 107.3% with coefficients of variation less than 14% demonstrating high assay accuracy and precision. Moreover, the actual sample analysis produced consistent results between this method and instrumental method. Therefore, the developed immunosensor can serve as an accurate and efficient approach for monitoring mycotoxins in agricultural products.

Topics: Animals; Antibodies; Cattle; Fluorescent Dyes; Immunoassay; Limit of Detection; Mycotoxins; Reproducibility of Results; Serum Albumin, Bovine; Spectroscopy, Near-Infrared; Trichothecenes; Zea mays; Zearalenone

Deoxynivalenol (DON) is found widely in foods and feeds that are contaminated with mildew and is one of the most harmful mycotoxins, threating not only human health but also impacting animal husbandry. Various physical, chemical and biological detoxification strategies have been applied in the past to reduce mycotoxin contamination. As a practical and economic method, addition of montmorillonite (Mt) offers the potential to eliminate mycotoxins, especially aflatoxin B1 (AFB

Topics: Adsorption; Aflatoxin B1; Bentonite; Hydrogen-Ion Concentration; Kinetics; Trichothecenes; Zearalenone

Topics: Animal Feed; Biotransformation; Chromatography, Liquid; Food Microbiology; Fusarium; Germany; Glucosides; Mass Spectrometry; Risk Assessment; Trichothecenes; Zea mays; Zearalenone; Zeranol

The immunotoxicity of zearalenone (ZEA) and deoxynivalenol (DON), two of the most common environmental mycotoxins, has been well investigated. However, due to the complexity of the immune system, especially during bacterial infection, many types of immune cells are involved in invasion resistance and bacterial clearance. Of these, T helper 2 (Th2) cells, which are members of the helper T cell family, assist B cells to activate and differentiate into antibody-secreting cells, participate in humoral immune response, and, ultimately, eliminate pathogens. Thus, it is important to identify the stage at which these toxins affect the immune function, and to clarity the underlying mechanisms. In this study, mice infected with Listeria monocytogenes (Listeria) were used to study the effects of ZEA, DON, and ZEA + DON on Th2 differentiation, Interleukin-4 Receptor (IL-4R) expression, costimulatory molecules expression and cytokine secretion after Listeria infection. Naive CD4

Topics: Animals; CD40 Ligand; Cell Differentiation; Cells, Cultured; Cytokines; Disease Models, Animal; Female; Host-Pathogen Interactions; Inducible T-Cell Co-Stimulator Protein; Listeria monocytogenes; Listeriosis; Lymphocyte Activation; Mice, Inbred BALB C; Mice, Inbred C57BL; Receptors, Interleukin-4; Signal Transduction; Spleen; Th2 Cells; Trichothecenes; Zearalenone

Toxic effects among fumonisins B (FB), deoxynivalenol (DON) and zearalenone (ZEN) administered alone and combined were investigated in 84-day-old ducks during force-feeding. 75 male ducks, divided into five groups of 15 animals, received daily during the meal a capsule containing the desired among of toxin. Treated animals received dietary levels of toxins equivalent to 20 mg FB1+FB2/kg (FB), 5 mg DON/kg (DON), 0.5 mg ZEN/kg (ZEN) and 20, 5 and 0.5 mg/kg of FB, DON and ZEN (FBDONZEN), respectively. Control birds received capsules with no toxin. After 12 days, a decrease in body weight gain accompanied by an increase in the feed conversion ratio was observed in ducks exposed to FBDONZEN, whereas there was no effect on performances in ducks exposed to FB, DON and ZEN separately. No difference among groups was observed in relative organ weight, biochemistry, histopathology and several variables used to measure oxidative damage and testicular function. A sphinganine to sphingosine ratio of 0.32, 1.19 and 1.04, was measured in liver in controls and in ducks exposed to FB and FBDONZEN, respectively. Concentrations of FB1 in liver were 13.34 and 15.4 ng/g in ducks exposed to FB and FBDONZEN, respectively. Together ZEN and its metabolites were measured after enzymatic hydrolysis of the conjugated forms. Mean concentrations of α-zearalenol in liver were 0.82 and 0.54 ng/g in ducks exposed to ZEN and FBDONZEN, respectively. β-zearalenol was 2.3-fold less abundant than α-zearalenol, whereas ZEN was only found in trace amounts. In conclusion, this study suggests that decreased performance may occur in ducks exposed to a combination of FB, DON and ZEN, but does not reveal any other interaction between mycotoxins in any of the other variables measured.

Topics: Animal Feed; Animal Husbandry; Animals; Biomarkers; Dietary Exposure; Ducks; European Union; Food Microbiology; Fumonisins; Humans; Kidney; Liver; Male; Maximum Tolerated Dose; No-Observed-Adverse-Effect Level; Organ Size; Risk Assessment; Trichothecenes; Weight Gain; Zearalenone

We assessed the mycobiota diversity and mycotoxin levels present in wild rice (Oryza latifolia) from the Pantanal region of Brazil; fundamental aspects of which are severely understudied as an edible plant from a natural ecosystem. We found multiple fungal species contaminating the rice samples; the most frequent genera being Fusarium, Nigrospora and Cladosporium (35.9%, 26.1% and 15%, respectively). Within the Fusarium genus, the wild rice samples were mostly contaminated by the Fusarium incarnatum-equiseti species complex (FIESC) (80%) along with Fusarium fujikuroi species complex (20%). Phylogenetic analysis supported multiple FIESC species and gave support to the presence of two putative new groups within the complex (LN1 and LN2). Deoxynivalenol (DON) and zearalenone (ZEN) chemical analysis showed that most of the isolates were DON/ZEN producers and some were defined as high ZEN producers, displaying abundant ZEN levels over DON (over 19 times more). Suggesting that ZEN likely has a key adaptive role for FIESC in wild rice (O. latifolia). Mycotoxin determination in the rice samples revealed high frequency of ZEN, and 85% of rice samples had levels >100 μg/kg; the recommended limit set by regulatory agencies. DON was only detected in 5.2% of the samples. Our data shows that FIESC species are the main source of ZEN contamination in wild rice and the excessive levels of ZEN found in the rice samples raises considerable safety concerns regarding wild rice consumption by humans and animals.

Topics: Animals; Brazil; Ecosystem; Food Contamination; Fusarium; Humans; Oryza; Phylogeny; Trichothecenes; Zearalenone

Deoxynivalenol (DON, vomitoxin) and Zearalenone (ZEA) are mycotoxin contaminants of cereals and cereal products that pose a significant threat to food safety. The aim of the study was to investigate the occurrence of DON and ZEA in different organic and conventional unprocessed cereals and cereal products that are available on the Polish agricultural fields and market. A total of 78 unprocessed cereal and cereal product samples of organic and conventional production were sampled from agricultural fields situated in western Poland and from available on the Polish market packaged comercial products produced by different domestic manufacturers. All samples were analyzed for DON and ZEA by HPLC with fluorescence detection (HPLC-FLD).. Results. Co-occurrence of DON was detected in cereals from the organic production system, the average content was 285.25 ± 134,04 μg kg. Mycotoxins contamination seen in organic cereals and cereal products does not statistical differ from that witnessed in their conventional counterparts.

Topics: Agriculture; Edible Grain; Flour; Food Contamination; Food, Organic; Organic Agriculture; Poland; Trichothecenes; Triticum; Zearalenone

Zearalenone (ZEA) and Deoxynivalenol (DON) are two mycotoxins highly detected in agricultural products and feed. Both mycotoxins produce reproductive toxicity and pose a serious threat to human and animal health, among which pigs are the most sensitive animals. Sertoli cells (SCs) play an important role in spermatogenesis; however, the combined toxicity of ZEA and DON and the screening of effective protective agents remains to be determined. By studying the effects of N-acetylcysteine (NAC) on the cells exposed to 20 μM of ZEA and 0.6 μM of DON, we explored the protective mechanism of NAC (4 mM) on the cytotoxic injury of piglets SCs induced by both mycotoxins. The results showed that the combination of ZEA and DON destroy organelles and SCs structures, NAC significantly alleviates the damage caused by ZEA and DON. NAC also significantly increased the expression and distribution of zonula occludens 1 (ZO-1), decreased the relative mRNA and protein expression levels of Bax, Bid, caspase-3, and caspase-9, and increased Bcl-2 expression level and inhibited the decrease of mitochondrial membrane potential. Further, NAC also eases the cell cycle arrest and oxidative stress caused by ZEA and DON. In summary, our results show that NAC could alleviate SCs injury via reducing the oxidative damage and apoptosis caused by ZEA and DON.

Topics: Acetylcysteine; Animals; Male; Sertoli Cells; Swine; Trichothecenes; Zearalenone

A microcosm study was conducted at two different temperatures under laboratory conditions to investigate the regulatory capacity and the interactive performance of two soil fauna species (Aporrectodea caliginosa, earthworms, and Proisotoma minuta, collembolans) on the reduction of Fusarium toxins in contaminated maize stubbles. Single and mixed species treatments were exposed to artificially infected maize stubbles highly contaminated with the mycotoxins deoxynivalenol (DON) (10,462 µg kg

Topics: Chromatography, Liquid; Food Contamination; Fusarium; Mass Spectrometry; Mycotoxins; Soil; Temperature; Trichothecenes; Zea mays; Zearalenone

Based on the fact that mycotoxins and the food-borne bacteria coexist in the natural environment and pose a significant health hazard to humans and animals, it is important to investigate the immunosuppressive mechanism of ZEA (zearalenone), DON (deoxynivalenol), and their combination in bacterial infections. In this study, we established a mouse model of mycotoxin low-dose exposure combined with Listeria monocytogenes infection and investigated the effects of ZEA, DON and their combination on Th1-mediated anti-intracellular bacterial infection based on CD4

Topics: Animals; CD4-Positive T-Lymphocytes; Cell Differentiation; Immunity, Cellular; Listeria monocytogenes; Mice; T-Lymphocytes; Trichothecenes; Zearalenone

Deoxynivalenol (DON), zearalenone (ZEN), and fumonisin B

Topics: Animals; Comet Assay; DNA Damage; Fumonisins; Male; Mycotoxins; Oryza; Rats; Rats, Sprague-Dawley; Trichothecenes; Zearalenone

Mycotoxins, toxins of fungal origin, can directly or indirectly contaminate food and feed and are poisonous to livestock and humans. While a large amount is known about their occurrence in crops, food, and feeds, little is known about mycotoxin amounts in soil. However, soil is known as a major fungal habitat and a potential sink for mycotoxins in the environment. Furthermore, there is neither a reliable detection nor an extraction method for mycotoxins testing in different soil textures or for potential deficits due to aging processes. Therefore, the aim of the present study was to present a reliable extraction and detection method for the simultaneous quantification of the most common mycotoxins, deoxynivalenol (DON) and zearalenone (ZEA), via liquid chromatography-tandem mass spectrometry (LC-MS/MS). This method was validated with six different samples with different textures and different soil organic matter (SOM). Deuterated standards were used to overcome possible matrix effects. This extraction method could eliminate potential aging processes. The recovery rate was always >80% for DON and >82% for ZEA. The quantification limits were 1 ng per g soil for DON and 0.5 ng per g soil for ZEA.

Topics: Chromatography, Liquid; Food Contamination; Humans; Mycotoxins; Soil; Soil Pollutants; Tandem Mass Spectrometry; Trichothecenes; Zea mays; Zearalenone

Topics: Crops, Agricultural; Edible Grain; Food Contamination; Food Handling; Fumonisins; Fusarium; Microwaves; Mycotoxins; Trichothecenes; Triticum; Zea mays; Zearalenone

Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins that contaminate a wide range of grains and crops. In this study, a one-step time-resolved single-channel immunochromatographic test strip based on europium ion polystyrene fluorescence microspheres was first developed for sensitive and quantitative detection of DON and ZEN. The concentration of the artificial antigen and the mass ratio of the monoclonal antibody to fluorescent microspheres for conjugation were optimized to simplify the sample addition process during immunochromatographic assay and improve the on-site detection efficiency. The limits of detection (LOD) of the single-channel immunochromatographic test strip for DON and ZEN detection were 0.17 and 0.54 μg/L, respectively. Meanwhile, the dual-channel immunochromatographic test strip was designed to simultaneously detect DON and ZEN, with LODs of 0.24 and 0.69 μg/L achieved for DON and ZEN, respectively. The developed test strips also yielded recovery results consistent with that obtained by LC-MS/MS for DON and ZEN detection in real samples of wheat and corn flour, confirming the practicability and reliability of the test strip. The developed immunochromatographic test strips realize quick and sensitive detection of DON and ZEN, exhibiting potential for broad applications in the point-of-care testing platform of multiple mycotoxins in agricultural products. Graphic abstract.

Topics: Edible Grain; Fluorescence; Immunoassay; Limit of Detection; Reagent Strips; Trichothecenes; Zea mays; Zearalenone

Human are exposed to a wide range of mycotoxins through dietary food intake, including processed food. Even most of the mycotoxin exposure assessment studies are based on analysis of foodstuffs, and evaluation of dietary intake through food consumption patterns and human biomonitoring methods are rising as a reliable alternative to approach the individual exposures, overcoming the limitations of the indirect dietary assessment. In this study, human urine samples were analyzed, seeking the presence of deoxynivalenol (DON), ochratoxin A (OTA), zearalenone (ZEA), and their metabolites. For this purpose, 40 urine samples from female and male adult residents in the city of Valencia (Spain) were evaluated by liquid chromatography quadrupole time-of-flight mass spectrometry (LC-ESI-qTOF) after salting-out liquid-liquid extraction. Analytical data showed that 72.5% of analyzed samples were contaminated by at least one mycotoxin at variable levels. The most prevalent mycotoxins were de-epoxy DON (DOM-1) (53%), ZEA (40%), and α-zearalenol (αZOL) (43%), while OTA was only detected in one sample. The mean concentrations in positive samples were DON (9.07 ng/mL), DOM-1 (20.28 ng/mL), ZEA (6.70 ng/mL), ZEA-14 glucoside (ZEA-14-Glc) (12.43 ng/mL), αZOL (27.44 ng/mL), αZOL-14 glucoside (αZOL-14-Glc) (12.84 ng/mL), and OTA (11.73 ng/mL). Finally, probable daily intakes (PDIs) were calculated and compared with the established tolerable daily intakes (TDIs) to estimate the potential risk of exposure to the studied mycotoxins. The calculated PDI was below the TDI value established for DON in both female and male adults, reaching a percentage up to 30%; however, this percentage increased up to 92% considering total DON (DON + DOM-1). On the other hand, the PDI obtained for ZEA and its metabolites were higher than the TDI value fixed, but the low urine excretion rate (10%) considered should be highlighted. Finally, the PDI calculated in the detected positive sample for OTA exceeded the TDI value. The findings of the present study confirm the presence of the studied mycotoxins and their metabolites as some of the most prevalent in urine.

Topics: Adolescent; Adult; Aged; Biological Monitoring; Biomarkers; Chromatography, Liquid; Dietary Exposure; Female; Humans; Male; Middle Aged; Ochratoxins; Risk Assessment; Spectrometry, Mass, Electrospray Ionization; Trichothecenes; Young Adult; Zearalenone

(1) Background: Deoxynivalenol (DON) and zearalenone (ZEA) are type B trichothecene mycotoxins that exert serious toxic effects on the reproduction of domestic animals. However, there is little information about the toxicity of mycotoxins on testis development in

Topics: Animals; Apoptosis; Cells, Cultured; Equidae; Gene Expression Regulation; Gene Regulatory Networks; Male; RNA, Messenger; Sertoli Cells; Transcriptome; Trichothecenes; Zearalenone

Fusarium mycotoxins and their derivatives are frequently detected in freshly harvested forage maize. This study assessed the time course effects during ensiling of forage maize on the fate of Fusarium mycotoxins, using laboratory-scale silos and artificially contaminated raw material. A multi-mycotoxin liquid chromatography-high-resolution mass spectrometry (LC-HRMS) method was used to determine the levels of deoxynivalenol (DON), zearalenone (ZEN) and their derivatives DON-3-glucoside, 3-acetyl-DON, 15-acetyl-DON, deepoxy-DON, α-zearalenol and β-zearalenol. A significant increase of DON was observed during ensiling, whereas the levels of DON-3-glucoside and its acetylated forms proportionally decreased. In contrast, levels of ZEN, α-zearalenol and β-zearalenol were not affected by the ensiling process. Based on these findings, ensiling is not a practical method for reducing the total amount of Fusarium mycotoxins present at harvest.

Topics: Chromatography, Liquid; Fusarium; Silage; Tandem Mass Spectrometry; Trichothecenes; Zea mays; Zearalenone

Plant-based materials used in the production of pig feed are very often contaminated with deoxynivalenol and zearalenone. Daily intake of small amounts of these mycotoxins with feed induces various subclinical states in gilts and influences different biological processes. The aim of this preclinical study was to determine the correlation between monotonic doses of zearalenone and deoxynivalenol (40 μg/kg body weight and 12 μg/kg body weight, respectively, administered over a period of 42 days) and the immunohistochemical expression of estrogen receptors in the intestinal tract and the mRNA expression of selected colonic enzymes. The immunohistochemical expression of estrogen receptor alpha was observed in the colon, but its intensity varied in different weeks of exposure. A minor increase in estrogen receptor beta expression was noted only in the colon, whereas the expression of cytochrome P450 1A1 enzyme mRNA and mRNA isoform of the glutathione S-transferase π gene decreased. The observed correlations suggest that the risk of loss of control over the biotransformation and biological activity of the parent compounds in distal intestinal mucosa is delayed.

Topics: Animals; Body Weight; Colon; Cytochrome P-450 CYP1A1; Estrogens, Non-Steroidal; Female; Glutathione Transferase; Intestinal Mucosa; Intestines; Mycotoxins; Poisons; Receptors, Estrogen; RNA, Messenger; Sus scrofa; Swine; Trichothecenes; Zearalenone

Fluorescent pseudomonads colonizing wheat ears have a high antagonistic potential against phytopathogenic fungi. To check this hypothesis, the bacterial antagonist Pseudomonas simiae 9

Topics: Alternaria; Antibiosis; Biological Control Agents; Fusarium; Lactones; Mycotoxins; Pseudomonas; Tenuazonic Acid; Trichothecenes; Triticum; Zearalenone

Topics: Aflatoxins; Biomarkers; Child; Child, Preschool; Diet; Environmental Exposure; Female; Food Contamination; Humans; Male; Mycotoxins; Ochratoxins; Surveys and Questionnaires; T-2 Toxin; Trichothecenes; United Kingdom; Zearalenone; Zeranol

A dietary exposure assessment to sum of deoxynivalenol (DON) forms, sum of T-2/HT-2 toxins (T2/HT2) and zearalenone (ZEA) was conducted for Czech children 4-6 years and Czech men and women 18-59 years. Retail foods (25 different commodities, n = 336) were assessed by LC-MS/MS methods. The 95th percentile chronic exposure to sum of DON forms was determined in children from 648 to 1030 ng/kg bw/day (LB/lower bound/and UB/upper bound/), in men from 362 to 923 ng/kg bw/day and in women from 272 to 490 ng/kg bw/day. The 95th percentile chronic exposure to sum T2/HT2 was determined in children from 6.5 to 31 ng/kg bw/day, in men from 1.9 to 11.2 ng/kg bw/day and in women from 2.5 to 11.5 ng/kg bw/day. The 95th percentile chronic exposure to ZEA was determined in children from 11.9 to 24.9 ng/kg bw/day, in men from 5.9 to 27.5 ng/kg bw/day and in women from 4.8 to 12.6 ng/kg bw/day. The risk linked with the mean and the 95th percentile chronic exposure (LB scenario) to the sum of DON forms, sum of T2/HT2 and ZEA is considered to be out of health concern for the selected population groups.

Topics: Adolescent; Adult; Beer; Child; Child, Preschool; Dietary Exposure; Edible Grain; Female; Food Contamination; Humans; Male; Middle Aged; Mycotoxins; T-2 Toxin; Trichothecenes; Young Adult; Zearalenone

A survey on 120 cereal samples (barley, maize, rice and wheat) from Algerian markets has been carried out to evaluate the presence of 15 mycotoxins (ochratoxin A, deoxynivalenol, fumonisin B1 and B2, T-2 and HT-2 toxins, zearalenone, fusarenon X, citrinin, sterigmatocystin, enniatins A, A1, B and B1, and beauvericin). With this purpose, a QuEChERS-based extraction and ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) were used. Analytical results showed that 78 cereal samples (65%) were contaminated with at least one toxin, while 50% were contaminated with three to nine mycotoxins. T-2 toxin, citrinin, beauvericin and deoxynivalenol were the most commonly found mycotoxins (frequency of 50%, 41.6%, 40.8% and 33.3%, respectively). Fumonisins (B1 + B2), enniatins B and B1, deoxynivalenol and zearalenone registered high concentrations (289-48878 µg/kg, 1.2-5288 µg/kg, 15-4569 µg/kg, 48-2055 µg/kg and 10.4-579 µg/kg, respectively). Furthermore, concentrations higher than those allowed by the European Union (EU) were observed in 21, 8 and 1 samples for fumonisins, zearalenone and deoxinivalenol, respectively. As a conclusion, the high levels of fumonisins (B1 + B2) in maize and deoxynivalenol, zearalenone and HT-2 + T-2 toxins in wheat, represent a health risk for the average adult consumer in Algeria. These results pointed out the necessity of a consistent control and the definition of maximum allowed levels for mycotoxins in Algerian foodstuffs.

Topics: Algeria; Chromatography, High Pressure Liquid; Dietary Exposure; Edible Grain; Food Contamination; Fumonisins; Humans; Limit of Detection; Maximum Allowable Concentration; Reproducibility of Results; Risk Assessment; Tandem Mass Spectrometry; Trichothecenes; Triticum; Zea mays; Zearalenone

Topics: Antibodies, Immobilized; Antibodies, Monoclonal; Cadmium Compounds; Food Contamination; Immunoassay; Luminescent Agents; Mycotoxins; Proof of Concept Study; Quantum Dots; Selenium Compounds; Sulfides; Trichothecenes; Triticum; Zea mays; Zearalenone; Zinc Compounds

Mycotoxins are a major contaminant of pig feed and have negative effects on health and performance. The present study investigated the impact of single or repeated acute challenges with a diet naturally contaminated with deoxynivalenol (DON) and zearalenone (ZEN) on growth performances of finishing pigs and their fecal microbiota composition. A total of 160 pigs (castrated males and females) in two successive batches were randomly divided into four experimental groups of 40 pigs each. The control group received a control finisher diet from 99 to 154 days of age. Challenged groups were subjected to a 7-day acute challenge by being fed a DON- and ZEN-contaminated diet (3.02 mg DON/kg feed and 0.76 mg ZEN/kg feed) at 113 days (group DC), 134 days (group CD) or both 113 and 134 days (group DD). Microbiota composition was analyzed via 16S rRNA sequencing from fecal samples collected from the 80 females at 99, 119, 140 and 154 days. Challenged pigs (i.e. groups DC, CD and DD) reduced their average daily feed intake by 25% and 27% (P < 0.001) and feed efficiency by 34% and 28% (P < 0.05) during the first and second mycotoxin exposure, respectively. Microbiota composition was affected by mycotoxin exposure (P = 0.07 during the first exposure and P = 0.01 during the second exposure). At the family level, mycotoxin exposure significantly (P < 0.05) decreased the relative abundances of Ruminococcaceae, Streptococcaceae and Veillonellaceae and increased that of Erysipelotrichaceae at both 119 and 140 days of age. After the 7-day DON/ZEN challenge, the relative abundance of 6 to 148 operational taxonomic units (OTUs) differed among the treatment groups. However, none of these OTUs changed in all treatment groups. Using 27 functional pathways, pigs exposed to DON/ZEN challenges could be distinguished from control pigs using sparse partial least squares discriminant analysis, with a 15% misclassification rate. Regarding the functionality of these predictors, two pathways were involved in detoxifying mycotoxins: drug metabolism and xenobiotic metabolism by cytochrome P450. In challenged pigs, microbiota composition returned to the initial state within 3 weeks after the end of a single or repeated DON/ZEN challenge, highlighting the resilience of the gut microbiome. The feeding and growth performances of the pigs during challenge periods were significantly correlated with biological pathways related to health problems and modifications in host metabolism. To conclude, sho

Topics: Animal Feed; Animals; Diet; Female; Food Contamination; Male; Microbiota; RNA, Ribosomal, 16S; Swine; Trichothecenes; Zearalenone

Currently there are few reports on the combined immunotoxicity of zearaleone (ZEA) and deoxynivalenol (DON). Since the two coexist naturally, it is necessary to understand the immunotoxicity caused by the two mycotoxins alone and in combination. To examine T lymphocytes activation and immune effect during activation, we used mouse primary spleen T lymphocytes as the experimental material and concanavalin (Con A) as the stimulator. The effects of ZEA, DON, and their combined exposure on T lymphocytes immune related function and the relationship between the activation of the mitogen-activated protein kinase (MAPK) signaling pathway and mycotoxin induced T lymphocytes apoptosis were studied in vitro. Specifically, T lymphocytes were isolated from primary mouse splenic lymphocytes, activated by Con A and then exposed to different concentrations of ZEA, DON, and their combinations. Our results showed that ZEA and DON alone and their combinations (20:1) can decrease the cell viability of T lymphocytes activated by Con A. The inhibitory effect of the combined groups was greater than that of the single mycotoxins, showing a synergistic effect. In addition, single or combined mycotoxins can lead to intracellular and surface ultrastructure damage of T lymphocytes, inhibit the expression of CD25 and CD278 and inhibit the synthesis of effect molecules poreforming protein (PFP), granzyme A (GZMA), and tumor necrosis factor-α (TNF-α). Meanwhile, the single mycotoxin or combined mycotoxins can promote the apoptosis of T lymphocytes which was accompanied by the overactivation of MAPK. After using the inhibitors of extracellular regulated protein kinases (ERK) and c-Jun N-terminal kinase (JNK) in the MAPK pathway, we found that the apoptosis of the cells induced by the ZEA was significantly decreased, and the apoptosis of the cells induced by DON had no significant changes. This suggests that the activation of MAPK induced by ZEA can promote the apoptosis of T lymphocytes, but the activation of MAPK induced by DON is not directly related to T cell apoptosis.

Topics: Animals; Apoptosis; Cell Survival; Immunotoxins; Interleukin-2 Receptor alpha Subunit; JNK Mitogen-Activated Protein Kinases; Mice; Mitogen-Activated Protein Kinases; Mycotoxins; Signal Transduction; T-Lymphocytes; Trichothecenes; Tumor Necrosis Factor-alpha; Zearalenone

The purpose of this study was to determine the effects of single and combined administrations of deoxynivalenol (DON) and zearalenone (ZEN) on the histology and ultrastructure of pig liver. The study was performed on immature gilts, which were divided into four equal groups. Animals in the experimental groups received DON at a dose of 12 μg/kg body weight (BW) per day, ZEN at 40 μg/kg BW per day, or a mixture of DON (12 μg/kg BW per day) and ZEN (40 μg/kg BW). The control group received vehicle. The animals were killed after 1, 3, and 6 weeks of experiment. Treatment with mycotoxins resulted in several changes in liver histology and ultrastructure, including: (1) an increase in the thickness of the perilobular connective tissue and its penetration to the lobules in gilts receiving DON and DON + ZEN; (2) an increase in the total microscopic liver score (histology activity index (HAI)) in pigs receiving DON and DON + ZEN; (3) dilatation of hepatic sinusoids in pigs receiving ZEN, DON and DON + ZEN; (4) temporary changes in glycogen content in all experimental groups; (5) an increase in iron accumulation in the hepatocytes of gilts treated with ZEN and DON + ZEN; (6) changes in endoplasmic reticulum organization in the hepatocytes of pigs receiving toxins; (7) changes in morphology of Browicz-Kupffer cells after treatment with ZEN, DON, and DON + ZEN. The results show that low doses of mycotoxins used in the present study, even when applied for a short period, affected liver morphology.

Topics: Animals; Chemical and Drug Induced Liver Injury; Female; Glycogen; Iron; Liver; Necrosis; Sus scrofa; Trichothecenes; Zearalenone

Mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) can negatively affect pig health. However, little is known about their effects on boar semen. We assessed the individual and combined effects of DON and ZEN on boar semen in vitro. In a pretrial, we determined the minimum dose (MiD) of each mycotoxin that induces a significant alteration of sperm progressive motility, as investigated using computer-assisted semen analysis (CASA). In the main trial, the individual and combined effects of each mycotoxin's MiD on sperm motility and kinetics (CASA analysis), morphology (SpermBlue staining), viability (calcein-propidium iodide staining), membrane functional status (hypoosmotic swelling test), and chromatin integrity (acridine orange staining) were analyzed. Pretrial results suggested a MiD of 50.6 μM and 62.8 μM for DON and ZEN, respectively. In the main trial, DON and ZEN administered at MiD significantly affected CASA parameters (e.g., increase of immotile spermatozoa, reduction of progressive motile spermatozoa), decreased sperm viability, and affected sperm morphology (head abnormalities) and membrane functional status. DON and ZEN showed less than additive effects on most parameters tested and a synergistic effect on viability and on two CASA parameters. In conclusion, DON and ZEN showed individual and combined toxic effects on boar semen in vitro.

Topics: Animals; Cell Survival; Drug Interactions; Male; Semen; Sperm Motility; Spermatozoa; Swine; Trichothecenes; Zearalenone

The intestinal epithelium is the first barrier against food contaminants and is highly sensitive to Fusarium toxins, especially deoxynivalenol (DON) and zearalenone (ZEA). Here, we explored the effects of low doses of DON and/or ZEA in naturally moldy diets on intestinal functions in piglets, including inflammatory responses, epithelial barrier, and microbial composition. Piglets were treated with a control diet (CON), DON diet (1000.6 μg/kg), ZEA diet (269.1 μg/kg), and DON + ZEA diet (1007.5 + 265.4 μg/kg), respectively, for 3 weeks and then switched to the same CON diet for another 2 weeks. In the first period, even the selected low doses of DON or ZEA in the diet resulted in intestinal inflammation, diminish protein expression (claudin-4) and altered gut microbiota populations. Whereas upon switching to the CON diet for another 2 weeks, the deleterious effect of ZEA and DON on IL-1β and Bifidobacterium population could not be recovered. Additionally, combined DON and ZEA negatively affected body weight gain and feed consumption of piglets, as well as shown synergistic effects on evoking pro-inflammatory cytokines contents (TNF-α, IL-1β, and IL-6) and perturbing the cecum microbiota profile (E. coli, Lactobacillus, and Bifidobacterium). Collectively, chronic consumption of DON and ZEA contaminated feed or food, even at low doses, can induce intestinal damage and may have consequences for animal and human health.

Topics: Animal Feed; Animals; Body Weight; Cecum; Cytokines; Diet; Dose-Response Relationship, Drug; Drug Synergism; Female; Food Contamination; Fusarium; Gene Expression; Hordeum; Immunity, Mucosal; Inflammation; Intestinal Mucosa; Jejunum; Swine; Tight Junction Proteins; Trichothecenes; Zea mays; Zearalenone

Topics: Animal Feed; Animals; Chickens; Diet; Dietary Supplements; Fumonisins; Liver; Male; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Turkeys; Zearalenone; Zeranol

Mycotoxin management in agriculture is an essential challenge for maintaining the health of both animals and humans. Choosing the right adsorbent is still a question for many breeders and an important criterion for feed manufacturers. New adsorbents are still being sought. Graphene oxide is a promising material in the field of nanotechnology, which excels in its adsorption properties. Presented in vitro study investigates graphene oxide for the binding of mycotoxins from crushed wheat. The results show that graphene oxide has an adsorption capacity for aflatoxin 0.045 mg/g, zearalenone 0.53 mg/g and deoxynivalenol 1.69 mg/g at 37° C. In vitro simulation of crushed wheat digestion showed rapid adsorption during the gastric phase. Of the minerals, Mg, Cu and Zn were the most adsorbed. The applied dose of graphene oxide of 10 mg/g caused only a slight inhibition of the digestive enzymes α-amylase and trypsin compared to pepsin and gastric lipase. In vitro results indicated the suitability of graphene oxide in the adsorption of the aflatoxin, zearalenone and deoxynivalenol.

Topics: Adsorption; Aflatoxin B1; Animal Feed; Animals; Digestion; Food Contamination; Gastrointestinal Absorption; Graphite; Humans; In Vitro Techniques; Mycotoxins; Nanostructures; Trichothecenes; Triticum; Zearalenone

Agricultural commodities, particularly cereals can be contaminated with mycotoxins during the pre- and post-harvest stage. The main goal of this study was to evaluate the efficacy of magnetic zeolite nanocomposite (MZNC) as an adsorbent for the reduction of mycotoxins in barley flour. The MZNC is synthesised using an eco-friendly and efficient procedure and characterised by zeta potential, field emission scanning electron microscopy, and energy-dispersive X-ray spectroscopy. The adsorbent amount that affects the adsorption capacity was optimised. Low amounts of the nanocomposite removed >99% of aflatoxins, 50% of ochratoxin A, 22% of zearalenone, and 1.8% of the deoxynivalenol from the contaminated sample and adsorption by MZNC was better than the natural zeolite; this phenomenon is related to the wide surface of nanocomposites. Results provide new insights into possible future research that could overcome the challenges of using nanotechnology to eliminate mycotoxins from agricultural products. It can be hoped that the presence of cheap and eco-friendly mycotoxin binders such as the MZNC that is synthesised and utilised in this research will help to produce secure food and feed products.

Topics: Adsorption; Aflatoxins; Centaurea; Edible Grain; Hordeum; Magnetic Phenomena; Microscopy, Electron, Scanning; Mycotoxins; Nanotechnology; Ochratoxins; Plant Extracts; Powders; Trichothecenes; Zearalenone; Zeolites

An analytical method for the simultaneous determination of trichothecenes-namely, nivalenol (NIV), deoxynivalenol (DON) and its acetylated derivatives (3- and 15-acetyl-DON), T-2 and HT-2 toxins-and zearalenone (ZEN) in wheat, wheat flour, and wheat crackers was validated through a collaborative study involving 15 participants from 10 countries. The validation study, performed within the M/520 standardization mandate of the European Commission, was carried out according to the IUPAC (International Union of Pure and Applied Chemistry) International Harmonized Protocol. The method was based on mycotoxin extraction from the homogenized sample material with a mixture of acetonitrile-water followed by purification and concentration on a solid phase extraction column. High-performance liquid chromatography coupled with tandem mass spectrometry was used for mycotoxin detection, using isotopically labelled mycotoxins as internal standards. The tested contamination ranges were from 27.7 to 378 μg/kg for NIV, from 234 to 2420 μg/kg for DON, from 18.5 to 137 μg/kg for 3-acetyl-DON, from 11.4 to 142 μg/kg for 15-acetyl-DON, from 2.1 to 37.6 μg/kg for T-2 toxin, from 6.6 to 134 μg/kg for HT-2 toxin, and from 31.6 to 230 μg/kg for ZEN. Recoveries were in the range 71-97% with the lowest values for NIV, the most polar mycotoxin. The relative standard deviation for repeatability (RSD

Topics: Chromatography, Liquid; Flour; Food Contamination; Humans; Intersectoral Collaboration; Mass Spectrometry; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Triticum; Whole Grains; Zearalenone

Mycotoxins are difficult to monitor continuously, and a tool to assess the risk would help to judge if there is a particular risk due to the inclusion of certain feed ingredients. For this, the toxin contents of 97 commercial fish feeds have been estimated, and the most prominent toxins in fish feed are calculated to be deoxynivalenol, zearalenone, fumonisins and enniatins. These pose a risk to fish well-being, as can be calculated by the Bayesian models for determining the critical concentrations 5% (CC5) for the different toxins. Besides fishmeal, wheat, soybean products and corn are regularly used as fish feed ingredients. The calculated scenarios show that fish are at high risk of toxin contamination if feed ingredients of low quality are chosen for feed production. Due to this, specific maximum allowable levels for several mycotoxins in fish feeds should be established.

Topics: Animal Feed; Animals; Aquaculture; Aspergillus; Fishes; Food Contamination; Fumonisins; Fusarium; Mycotoxins; Ochratoxins; Penicillium; Risk Assessment; Trichothecenes; Triticum; Zea mays; Zearalenone

Enzymatic treatment is an attractive method for mycotoxin detoxification, which ideally prefers the use of one or a few enzymes. However, this is challenged by the diverse structures and co-contamination of multiple mycotoxins in food and feed. Lignin-degrading fungi have been discovered to detoxify organics including mycotoxins. Manganese peroxidase (MnP) is a major enzyme responsible for lignin oxidative depolymerization in such fungi. Here, we demonstrate that eight MnPs from different lignocellulose-degrading fungi (five from

Topics: Aflatoxin B1; Food Contamination; Fumonisins; Fungi; Malonates; Metabolic Detoxication, Phase I; Mycotoxins; Naphthalenesulfonates; Peroxidases; Trichothecenes; Zearalenone

Topics: Animal Feed; Animals; Chickens; Diet; Fumonisins; Liver; Male; Muscles; Trichothecenes; Turkeys; Zearalenone

Mycotoxin removers include enzymes and adsorbents that may be used in animal feeds to eliminate the toxic effects of mycotoxins. This study aimed to determine the removability of two different types of mycotoxin removers, adsorbents and enzyme degradation reagents (EDRs), in the simulated gastrointestinal conditions of pigs and poultry. Seven commercial mycotoxin removers, including five EDRs and two adsorbents, were tested in vitro. In this study, the supplemented dosages of mycotoxin removers used in pig and poultry feeds were the commercial recommendation ranging from 0.05% to 0.2%. For pigs, the in vitro gastric and small intestinal simulations were performed by immersing the mycotoxin-tainted feed in artificial gastric juice (AGJ) at pH 2.5 for 5 h or in artificial intestinal juice (AIJ) at pH 6.5 for 2 h to mimick in vivo conditions. For poultry, mycotoxin-tainted feeds were immersed in AGJ for 2 h at pH 4.5 and 0.5 h at pH of 2.5, respectively, to simulate crop/glandular stomach and gizzard conditions; the small intestinal simulation was in AIJ for 2 h at pH 6.5. For the pig, EDRs and adsorbents had deoxynivalenol (DON) removability (1 mg/kg) of 56% to 100% and 15% to 19%, respectively. Under the concentration of 0.5 mg/kg, the zearalenone (ZEN) removability by EDRs and adsorbents was 65% to 100% and 0% to 36%, respectively. For the simulation in poultry, the removability of DON by EDRs and adsorbents (5 mg/kg) was 56% to 79% and 1% to 36%, respectively; for the concentration of 0.5 mg/kg, the removability of ZEN by EDRs and adsorbents was 38% to 69% and 7% to 9%, respectively. These results suggest that EDRs are more effective in reducing DON and ZEN contamination compared to the adsorbent methods in the simulated gastrointestinal tracts of pig and poultry. The recoveries of DON and ZEN of pig in vitro gastrointestinal simulations were higher than 86.4% and 84.7%, respectively, with 88.8% and 85.9%, respectively, in poultry. These results demonstrated the stability and accuracy of our mycotoxin extraction process and in vitro simulation efficiency.

Topics: Adsorption; Aluminum Silicates; Animals; Bacillus subtilis; Carboxylesterase; Carboxypeptidases; Cell Wall; Food Contamination; Gastric Juice; Hydrolases; Indicators and Reagents; Oxidoreductases; Poultry; Swine; Trichothecenes; Yeasts; Zearalenone

The aim of this study was to evaluate the effect of 15 commercial yeasts in the mitigation of the Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) during the brewing process. Saccharomyces strains (10 strains of S. cerevisiae and 5 of S. pastorianus) were used to ferment DON and ZEN contaminated wort. Wort samples were taken every 24 h during fermentation, while mycotoxin analysis in yeast was performed at the end of fermentation (96 h); additionally, pH and ethanol content were measured daily. For mycotoxin analysis, after immunoaffinity purification of sample extracts, analysis was performed using an Ultra-High-Pressure Liquid Chromatograph coupled with a diode array or fluorescence detector (UHPLC-DAD/FLD). Mycotoxin presence had no significant effect on the ethanol production during brewing. At the end of fermentation, 10-17% of DON and 30-70% of ZEN had been removed, 6% of the initial concentration of DON and 31% of the ZEN being adsorbed by the yeast. Beermakers must pay careful attention to the raw material since a high percentage of DON could be present at the end of the beer fermentation process. Future studies should focus on the quantification of "masked" mycotoxins that are relevant to food security.

Topics: Beer; Chromatography, High Pressure Liquid; Fermentation; Food Analysis; Food Contamination; Food Microbiology; Fusarium; Hydrogen-Ion Concentration; Mycotoxins; Reproducibility of Results; Saccharomyces; Saccharomyces cerevisiae; Trichothecenes; Zearalenone

Topics: Animal Feed; Animals; Diet; European Union; Food Contamination; Fumonisins; Mycotoxins; Trichothecenes; Turkey; Turkeys; Zearalenone

Topics: Aflatoxins; China; Dietary Exposure; Food Contamination; Food Safety; Fumonisins; Humans; Legislation, Food; Maximum Allowable Concentration; Mycotoxins; Trichothecenes; Zea mays; Zearalenone

In this study, 120 silage samples collected in 2015 from farms in Poland were analysed by a multimycotoxin method based on liquid chromatography coupled with tandem mass spectrometry. The study included toxins which are regulated within the European Union (aflatoxins, deoxynivalenol, fumonisins, T-2/HT-2 toxins, ochratoxin A and zearalenone) and non-regulated mycotoxins (enniatins, beauvericin, 8-ketotrichothecenes, sterigmatocystin, zearalenone derivatives). All silage samples were positive for at least one mycotoxin, and 61% of samples contained five or more mycotoxins simultaneously. The most frequently detected toxins were deoxynivalenol, nivalenol, zearalenone, enniatins and beauvericin, although the levels of these toxins were relatively low. The mean concentration of deoxynivalenol and zearalenon was 406 and 80.6 μg/kg, respectively, and two toxins were positive-correlated. This is the first study that provides information about emerging mycotoxins contaminating silage in Poland.

Topics: Chromatography, Liquid; Fumonisins; Fusarium; Mycotoxins; Poland; Silage; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

To observe the variation in accumulation of Fusarium and Alternaria mycotoxins across a topographically heterogeneous field and tested biotic (fungal and bacterial abundance) and abiotic (microclimate) parameters as explanatory variables.. We selected a wheat field characterized by a diversified topography, to be responsible for variations in productivity and in canopy-driven microclimate. Fusarium and Alternaria mycotoxins where quantified in wheat ears at three sampling dates between flowering and harvest at 40 points. Tenuazonic acid (TeA), alternariol (AOH), alternariol monomethyl ether (AME), tentoxin (TEN), deoxynivalenol (DON), zearalenone (ZEN) and deoxynivalenol-3-Glucoside (DON.3G) were quantified. In canopy temperature, air and soil humidity were recorded for each point with data-loggers. Fusarium spp. as trichothecene producers, Alternaria spp. and fungal abundances were assessed using qPCR. Pseudomonas fluorescens bacteria were quantified with a culture based method. We only found DON, DON.3G, TeA and TEN to be ubiquitous across the whole field, while AME, AOH and ZEN were only occasionally detected. Fusarium was more abundant in spots with high soil humidity, while Alternaria in warmer and drier spots. Mycotoxins correlated differently to the observed explanatory variables: positive correlations between DON accumulation, tri 5 gene and Fusarium abundance were clearly detected. The correlations among the others observed variables, such as microclimatic conditions, varied among the sampling dates. The results of statistical model identification do not exclude that species coexistence could influence mycotoxin production.. Fusarium and Alternaria mycotoxins accumulation varies heavily across the field and the sampling dates, providing the realism of landscape-scale studies. Mycotoxin concentrations appear to be partially explained by biotic and abiotic variables.. We provide a useful experimental design and useful data for understanding the dynamics of mycotoxin biosynthesis in wheat.

Topics: Alternaria; Food Contamination; Fusarium; Glucosides; Lactones; Microclimate; Mycotoxins; Pseudomonas fluorescens; Secondary Metabolism; Soil Microbiology; Tenuazonic Acid; Trichothecenes; Triticum; Zearalenone

Topics: Edible Grain; Food Contamination; Fusarium; Hordeum; Phylogeny; Trichothecenes; Zearalenone

Metabolic profiling in Caco-2 cells was studied for the combined toxic effects of deoxynivalenol (DON), zearalenone (ZEN), and Aflatoxin B

Topics: Aflatoxin B1; Apoptosis; bcl-2-Associated X Protein; Caco-2 Cells; Caspase 3; Caspase 8; Cell Survival; Humans; Metabolic Networks and Pathways; Trichothecenes; Tumor Suppressor Protein p53; Zearalenone

Mycotoxins are widely studied by many research groups in all aspects, but the stability of these compounds needs further research for clarification. The objective of this study is to evaluate deoxynivalenol and zearalenone stability during all steps of the malting and brewing processes. The levels of these compounds decreased significantly during the production process (barley to beer). During the malting process, the DON levels decreased significantly in the steeping, germination, and malting steps (62%, 51.5%, and 68%, respectively). Considering ZEN, when the levels were compared between barley and the last step of the process, a significant decrease was observed. Most of the mycotoxins produced were transferred to the rootlets and spent grains, which is advantageous considering the final product. Furthermore, the mycotoxin dietary intake estimation was included in this study. The results proved that if the concentrations of target mycotoxins in raw material are under the limits established by the regulations, the levels decrease during the malting and brewing processes and make the beer secure for consumers. The quality of the five commodities involved in the beer process plays a decisive role in the creation of a safe final product.

Topics: Adult; Beer; Dietary Exposure; Food Contamination; Food Industry; Fusarium; Hordeum; Humans; Trichothecenes; Zearalenone

Contamination of feed by mycotoxins is a global epidemic that has a sizeable impact on animal health and causes economic losses. Mycotoxins, including aflatoxins (AFs), zearalenone (ZEN), fumonisins (FUMs), deoxynivalenol (DON), and ochratoxin A (OTA), lead to acute and chronic adverse effects in pigs. Animal feed and feed ingredients are commonly contaminated by one or more mycotoxins worldwide; however, the prevalence of mycotoxin contamination in feed and feed ingredients in Taiwan remains unclear. A total of 820 cornmeal and corn-based swine feed (pregnancy and nursery diets) samples provided by feed and animal producers were analyzed using the enzyme-linked immunosorbent assay method between January 2015 and December 2017 to determine the presence of mycotoxins. The results revealed that the most prevalent mycotoxin in Taiwan was DON, with 91.4% of positive samples between 2015 and 2017, followed by ZEN, AFs, and FUMs, with 70.2%, 58.0%, and 50.4% of positive samples, respectively. A similar prevalence of mycotoxins was observed in cornmeal and corn-based swine feed. Furthermore, 7.7% of the analyzed feed samples contained one mycotoxin, and 91.3% contained multiple mycotoxins. DON was the most prevalent mycotoxin in cornmeal and corn-based swine feed in Taiwan. Moreover, a high incidence of contamination by multiple mycotoxins was observed in swine feed. Awareness of mycotoxin presence in feed and development of mycotoxin detoxification strategies are unmet needs.

Topics: Aflatoxins; Animal Feed; Animals; Diet; Enzyme-Linked Immunosorbent Assay; Food Contamination; Fumonisins; Mycotoxins; Ochratoxins; Swine; Taiwan; Trichothecenes; Zea mays; Zearalenone

The fungus Fusarium graminearum is the causative agent of economically significant plant diseases such as Fusarium Healed Blight (FHB) of cereals, its mycotoxins as deoxynivalenol (DON), Nivalenol (NIV) and Zearalenone (ZEN) contaminate wheat and other grains. The objectives of the present study were to determine the mechanism by which the bacterium Pseudomonas aeruginosa inhibits the growth of F. graminearum. Our results indicate that P. aeruginosa metabolites as pyocyanin has effective antifungal properties. Pyocyanin was produced by P. aeruginosa when cultured on mineral salt medium and reached a maximum concentration after 72 h. Pyocyanin significantly decreased mycotoxins of F. graminearum, a 25 mg/ml of pyocyanin for 72 h decreased DON by 68.7% and NIV by 57.7%.Real-Time PCR analysis demonstrated that the antifungal effect is mediated by downregulation of the Pleiotropic Drug Resistance (PDR) subfamily FgABC3. 25 mg/ml of pyocyanin decreased FgABC3-mRNA by 60%, inhibited the fungal growth and decreased the area of mycelial growth at 12, 24, 36 and 72 h post incubation by 40-50%. Deletion of FgABC3 led to enhanced accumulation of DON and NIV by 40 and 60%, respectively.The data presented in this report may have significance in understanding mechanism by which certain bacterial metabolites exert a beneficial effect and for developing antifungal drugs.

Topics: Antifungal Agents; Drug Resistance; Edible Grain; Fusarium; Mycotoxins; Pseudomonas aeruginosa; Pyocyanine; Trichothecenes; Triticum; Zearalenone

Mycotoxins are toxic metabolites produced by fungi or molds, which may cause serious harm to human health through polluted cereal foods. In order to measure the typical mycotoxin contaminations in wheat and corn, a surface plasmon resonance (SPR) method was established using SPR sensor chip that was fabricated based on self-assembled monolayer. The minimum detection limit of aflatoxin B1, ochratoxin A, zearalenone and deoxynivalenol were identified as 0.59 ng/mL, 1.27 ng/mL, 7.07 ng/mL and 3.26 ng/mL, respectively. The cross-reactivity for all four mycotoxins were demonstrated to be low. Moreover, the test data were compared with HPLC-MS/MS confirmatory analysis results and good agreement was found between them. In conclusion, the SPR method for simultaneously detecting four mycotoxins has been developed with high sensitivity, good linearity and specificity, which can meet the detection requirements of cereal foods.

Topics: Aflatoxin B1; Chromatography, High Pressure Liquid; Cross Reactions; Food Analysis; Food Contamination; Hydrazones; Limit of Detection; Mycotoxins; Ochratoxins; Sensitivity and Specificity; Surface Plasmon Resonance; Tandem Mass Spectrometry; Trichothecenes; Triticum; Zea mays; Zearalenone

Numerous surveys have highlighted the natural co-occurrence of deoxynivalenol (DON) and zearalenone (ZEA) mycotoxins in food and feed. Nevertheless, data regarding cellular mechanisms involved in response to their individual and simultaneous exposures are lacking. In this study, in order to analyze how low mycotoxin doses could impact cellular physiology and homeostasis, proteomic profiles of proliferating human hepatic cells (HepaRG) exposed for 1h and 24h to low DON and ZEA cytotoxicity levels (0.2 and 20μM respectively), alone or in combination, were analyzed by LC-MS/MS. Proteome analyses of mycotoxin-treated cells identified 4000 proteins with about 1.4% and 3.7% of these proteins exhibiting a significantly modified abundance compared to controls after 1h or 24h, respectively. Analysis of the Gene Ontology biological process annotations showed that cell cycle, proliferation and/or development as well as on DNA metabolic processes were affected for most treatments. Overall, different proteins, and thus biological processes, were impacted depending on the considered mycotoxin and exposure duration. Finally, despite the important proteome changes observed following 24h exposure to both mycotoxins, only the uptake of ZEA by the cells was suggested by the mycotoxin quantification in cell supernatants.. This study investigated the proteomic changes that occurred after DON and ZEA (individually and in combination) short exposures at low cytotoxicity levels in proliferating HepaRG cells using LC-MS/MS. The obtained results showed that the cellular response is time- and mycotoxin or mixture-dependent. In particular, after 1h exposure, the DON+ZEA combination led to more proteomic changes than DON or ZEA alone, whereas the opposite was observed after 24h. In addition, the significant cellular response to stress induced by ZEA after 24h exposure seemed to be reduced when combined with DON. Thus, these results supported a possible mitigation by the hepatocytes when exposed to the mycotoxin mixture for a long duration. These findings represent an essential step to further explore adaptive cell response to mycotoxin exposure using with more complex incubation kinetics and combining different "omics" tools. Moreover, as mycotoxin quantification in cell supernatants showed different behaviors for DON and ZEA, this also raises the question about how mycotoxins actually trigger the cell response.

Topics: Cell Cycle; Cell Proliferation; DNA; Drug Interactions; Environmental Exposure; Hepatocytes; Humans; Mycotoxins; Proteome; Trichothecenes; Zearalenone

Deoxynivalenol (DON) and zearalenone (ZEA) are mycotoxins primarily produced by Fusarium species and commonly co-occur in European grains. Some in vitro studies reported synergistic combined effects on cell viability reduction for these two natural food contaminants. However, most of these studies were carried out on conventional cell culture systems involving only one cell type and thus did not include cell-cell communication that is closer to in vivo conditions. In this context, we developed easy bi- and tri-culture systems using the Caco-2 (intestinal epithelial cells), THP-1 (monocytes) and HepaRG (hepatic cells) human cell lines in a proliferating state. Individual and combined cytotoxic effects of DON and ZEA were then assessed using co-cultures during 48 h. In bi-culture systems, results showed that only the highest tested dose of ZEA (IC

Topics: Cell Communication; Cell Line; Cell Survival; Chromatography, High Pressure Liquid; Coculture Techniques; Humans; Mycotoxins; Spectrometry, Mass, Electrospray Ionization; Toxicity Tests, Acute; Trichothecenes; Zearalenone

This paper presents data on the occurrence of Fusarium toxins - zearalenone (ZEA), deoxynivalenol (DON) and fumonisins (FUMs) B1 and B2 - in corn flours and corn flakes marketed in Serbia. A total of 71 samples were collected over 2013-2016 and analysed using HPLC with UV or fluorescence detection. In the case of corn flours, none of the samples taken in 2013 exhibited the presence of ZEA or DON, whereas 90% were positive for FUMs. In 2015, occurrence was very high: ZEA 93%, DON 86% and FUMs 100% (mean 43.3, 322.6 and 323.0 μg kg

Topics: Commerce; Food Analysis; Food Contamination; Fumonisins; Fusarium; Serbia; Trichothecenes; Zea mays; Zearalenone

A feeding experiment with piglets was performed to examine the efficacy of a wet preservation of Fusarium (FUS)-contaminated maize with sodium sulphite (SoS) based on deoxynivalenol (DON) and zearalenone (ZEN) residue levels in urine, bile and liquor and health traits of piglets. For this purpose, 80 castrated male piglets (7.57 ± 0.92 kg BW) were assigned to four treatment groups: CON- (control diet, with 0.09 mg DON and <0.01 mg ZEN/kg diet), CON+ (diet CON-, wet-preserved with 5 g SoS/kg maize; containing 0.05 mg DON and <0.01 mg ZEN/kg diet), FUS- (diet with mycotoxin-contaminated maize; containing 5.36 mg DON and 0.29 mg ZEN/kg diet), and FUS+ (diet FUS-, wet-preserved with 5 g SoS/kg maize; resulting in 0.83 mg DON and 0.27 mg ZEN/kg diet). After 42 d, 40 piglets (n = 10 per group) were sampled. A clear reduction of DON levels by approximately 75% was detected in all specimens of pigs fed diet FUS+. ZEN was detected in all urine, bile and liquor samples, while their metabolites were only detectable in urine and bile. Additionally, their concentrations were not influenced by SoS treatment. Among the health-related traits, feeding of FUS diets increased the total counts of leukocytes and segmented neutrophil granulocytes irrespective of SoS treatment. SoS treatment increased the total blood protein content slightly with a similar numerical trend in albumin concentration. These effects occurred at an obviously lower level in FUS-fed groups. Moreover, SoS treatment recovered the reduction of NO production induced by feeding diet FUS- indicating an effect on the redox level. As this effect only occurred in group FUS+, it is obviously related to the adverse effects of the Fusarium toxins. In conclusion, treatment of FUS-contaminated maize with SoS decreased the inner exposure with DON as indicated by the lower DON levels in various piglet specimens. However, health-related traits did not consistently reflect this decreased exposure.

Topics: Animal Feed; Animals; Decontamination; Diet; Fusarium; Male; Mycotoxins; Random Allocation; Sulfites; Sus scrofa; Trichothecenes; Zea mays; Zearalenone

Wheat bran is an important source for human and animal feed. Its nutritional aspects include a high content of fibre and minerals, as well as phenolic compounds that help prevent chronic diseases. However, wheat can be susceptible to contamination by fungus, which can produce mycotoxins such as deoxynivalenol (DON) and zearalenone (ZEN), causing adverse health effects. Therefore, methods should be developed to reduce possible contamination. Ozone can be used for this purpose as it is considered safe and environmental friendly. The aim of this study was to evaluate the reduction of DON and ZEN concentrations in wheat bran using the ozonation process as well as to evaluate the effect of ozonation on the nutritional quality of bran. Considering this, wheat bran naturally contaminated with both DON and ZEN was processed using ozone at different conditions. The nutritional quality of the bran was evaluated after processing considering the following aspects: the total phenolic content and the bran antioxidant capacity (by using both DPPH and ABTS radicals). The results showed that the degradation of ZEN was higher and faster than the degradation of DON, which could be explained by their molecular structures. The total phenolic content and antioxidant capacity of the bran were not affected by the ozonation process, which is preferable from a nutritional point of view. Therefore, ozonation was demonstrated to be a possible method for reducing mycotoxins in wheat bran, although more studies are needed in order to better understand and optimise processing and product quality.

Topics: Dietary Fiber; Food Contamination; Nutritive Value; Ozone; Trichothecenes; Zearalenone

Mycotoxins produced by different species of fungi may coexist in cereals and feedstuffs, and could be highly toxic for humans and animals. For quantification of multiple mycotoxins in cereals, we developed a paper-based mycotoxin immune-affinity array. First, paper-based microzone arrays were fabricated by photolithography. Then, monoclonal mycotoxin antibodies were added in a copolymerization reaction with a cross-linker to form an immune-affinity monolith on the paper-based microzone array. With use of a competitive immune-response format, paper-based mycotoxin immune-affinity arrays were successfully applied to detect mycotoxins in samples. The detection limits for deoxynivalenol, zearalenone, T-2 toxin, and HT-2 toxin were 62.7, 10.8, 0.36, and 0.23 μg·kg

Topics: Animals; Antibodies, Immobilized; Edible Grain; Equipment Design; Food Analysis; Food Contamination; Humans; Immunoassay; Limit of Detection; Luminescent Measurements; Mycotoxins; Paper; Polymerization; Reproducibility of Results; T-2 Toxin; Trichothecenes; Zearalenone

Urine metabolic profiling of mice was conducted utilizing gas chromatography-mass spectrometry (GC-MS) to investigate the combinatory effect of mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) on the metabolism of the mice. Experiments were conducted by means of five-week-old mice which were individually exposed to 2 mg/kg DON, 20 mg/kg ZEN and the mixture of DON and ZEN (2 mg/kg and 20 mg/kg, respectively). The intragastric administration was applied for three weeks and urine samples were collected for metabolic analysis. Univariate and multivariate analysis were applied to data matrix processing along with respective pathway analysis by MetaMapp and CytoScape. The results showed that the combined DON and ZEN administration resulted in lower significant changes, compared to the individual mycotoxin treated groups verified by heatmap. Metabolic pathways network mapping indicated that the combined mycotoxins treated groups showed a little effect on the metabolites in most pathways, especially in glucose metabolism and its downstream amino acid metabolism. In glucose metabolism, the content of galactose, mannitol, galactonic acid, myo-inositol, tagatose was drastically down-regulated. Furthermore, the organic acids, pyruvate, and amino acids metabolism displayed the same phenomenon. In conclusion, the combined DON/ZEN administration might lead to an "antagonistic effect" in mice metabolism.

Topics: Animals; Drug Information Services; Female; Gas Chromatography-Mass Spectrometry; Male; Metabolomics; Mice; Multivariate Analysis; Trichothecenes; Urinalysis; Zearalenone

The objective of this study was to investigate the individual and combined contamination of aflatoxin B₁ (AFB₁), zearalenone (ZEN) and deoxynivalenol (DON) in feedstuffs from different Provinces of China between 2016 and 2017. A total of 1569 samples, including 742 feed ingredients and 827 complete pig feed samples, were collected from various regions of China for mycotoxins analysis. The results showed that individual occurrence rates of AFB₁, ZEN, and DON were more than 83.3%, 88%, and 74.5%, respectively, in all the tested samples. DON was the most prevalent contaminant, followed by ZEN and AFB₁, with the average concentrations ranging from 450.0-4381.5 μg/kg, 2.3-729.2 μg/kg, and 1.3-10.0 μg/kg, respectively. Notable, 38.2%, 10.8%, and 0.6% of complete pig feeds were contaminated with DON, ZEN, and AFB₁ over China's regulatory limits, respectively. Moreover, over 75.0% analyzed samples were co-contaminated with two or three mycotoxins. In conclusion, the current study revealed that the feedstuffs in China were severely contaminated with DON, followed by ZEN and AFB₁ during the past two years. These findings highlight the importance of monitoring mycotoxins in livestock feed and implementing feed management and bioremediation strategies to reduce mycotoxin exposure.

Topics: Aflatoxin B1; Animal Feed; China; Environmental Monitoring; Food Contamination; Trichothecenes; Zearalenone

Barley (Hordeum vulgare L.) is an important cereal crop for food and represents one of the main ingredients in beer production. Considering the importance of barley and its derived products, the knowledge about the mycotoxin contamination in the barley production is essential in order to assess its safety. In this study, the levels of deoxynivalenol (DON) and zearalenone (ZEN) in brewing barley were determined using a LC-MS/MS method. A survey was conducted in 2015 to estimate the mycotoxin levels in these products (n = 76) from four crop regions in Brazil. The results showed high levels of DON and ZEN in the analyzed samples, with contamination levels of 94 and 73.6%, respectively. The mean levels of DON and ZEN ranged from 1700 to 7500 μg/kg and from 300 to 630 μg/kg, respectively. Barley samples from regions 1 and 2 presented higher levels of ZEN and DON, respectively, and those from region 4 presented lower levels of both. Co-occurrence of DON and ZEN was seen in the majority of the barley grain samples, and the mycotoxin content was above the maximum levels established by the Brazilian and European regulations.

Topics: Beer; Brazil; Chromatography, Liquid; Food Contamination; Hordeum; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

The contamination of feed with mycotoxins results in reduced growth, feed refusal, immunosuppression, and health problems. Deoxynivalenol (DON) and zearalenone (ZEN) are among the most important mycotoxins. The aim of the study was to examine the effects of low doses of these mycotoxins on the histological structure and ultrastructure of the large intestine in the pig. The study was performed on 36 immature gilts of mixed breed (White Polish Big × Polish White Earhanging), which were divided into four groups administrated per os with ZEN at 40 µg/kg BW, DON at 12 µg/kg BW, a mixture of ZEN (40 µg/kg BW) and DON (12 µg/kg BW) or a placebo. The pigs were killed by intravenous overdose of pentobarbital after one, three, and six weeks of treatment. The cecum, ascending and descending colon samples were prepared for light and electron microscopy. Administration of toxins did not influence the architecture of the mucosa and submucosa in the large intestine. ZEN and ZEN + DON significantly decreased the number of goblet cells in the cecum and descending colon. The mycotoxins changed the number of lymphocytes and plasma cells in the large intestine, which usually increased in number. However, this effect differed between the intestine segments and toxins. Mycotoxins induced some changes in the ultrastructure of the mucosal epithelium. They did not affect the expression of proliferative cell nuclear antigen and the intestinal barrier permeability. The obtained results indicate that mycotoxins especially ZEN may influence the defense mechanisms of the large intestine.

Topics: Animals; Female; Intestine, Large; Microscopy; Swine; Trichothecenes; Zearalenone

Topics: Beer; Food Contamination; Food Handling; Fumonisins; Hot Temperature; Nigeria; Spices; T-2 Toxin; Trichothecenes; Zearalenone

In this report, a UPLC-ESI-MS/MS method for the simultaneous determination of aflatoxins, ochratoxin A, zearalenone, deoxynivalenol, fumonisins, T-2 and HT-2 toxins, fusarenone X, diacetoxyscirpenol, and 3- and 15-acetyldeoxynivalenol in feedstuffs was developed. A quadrupole-time-of-flight mass spectrometer detector (QTOF-MS) operating in full scan mode was combined with the UPLC-ESI-MS/MS system to confirm the identity of detected mycotoxins and to identify other possible microbial metabolites occurring in samples. Sixty-two feed samples from the Spanish market were analyzed. Extraction of metabolites was carried out with acetonitrile-water-formic acid (80:19:1, v/v/v). Method detection and quantification limits and performance criteria set by Commission Regulation (EC) No 401/2006 were fulfilled. Relatively high levels of the main regulated mycotoxins and presence of non-regulated mycotoxins in feed samples were found. This is the first study in which mycotoxins and other microbial metabolites occurring in feed are studied using a UPLC-QTOF-MS system being therefore a reference report.

Topics: Aflatoxins; Animal Feed; Chromatography, High Pressure Liquid; Fumonisins; Mass Spectrometry; Mycotoxins; Ochratoxins; T-2 Toxin; Trichothecenes; Zearalenone

Topics: Animals; Behavior, Animal; Food Contamination; Larva; Mycotoxins; Ochratoxins; Trichothecenes; Water Pollutants, Chemical; Zearalenone; Zebrafish; Zeranol

Most plant materials are contaminated with small doses of Fusarium mycotoxins and its modified forms that exert subclinical toxic effects on humans and animals. The aim of this study was to evaluate the carry-over of zearalenone and deoxynivalenol (pure parent compounds) to intestinal and liver tissues during 6 weeks of exposure to mycotoxins administered per os to gilts. The experiment was performed on 36 gilts with average body weight of 25 ± 2 kg, divided into 2 groups: an experimental group (group E, administered zearalenone at 40 μg/kg BW and deoxynivalenol at 12 μg/kg BW daily with feed) and a control group administered placebo. Tissue saturation with mycotoxins was analysed by liquid chromatography in samples collected at weekly intervals. Six gilts were euthanized in each week of the study. The conducted analyses revealed: (i) a non-uniform increase in zearalenone levels in the duodenum, jejunum, ascending colon and the liver; and (ii) an increase in deoxynivalenol levels, mainly in the ileum, caecum, ascending colon and the transverse colon, and a minor increase in the liver. The degree of tissue saturation was determined by the type of mycotoxin, but not by the time of exposure.

Topics: Animal Feed; Animals; Chromatography, Liquid; Female; Food Contamination; Intestinal Mucosa; Intestines; Liver; Swine; Trichothecenes; Zearalenone

Deoxynivalenol (DON) and zearalenone (ZEN) can seriously affect animal health, with potentially severe economic losses. Previous studies have demonstrated that gut microbiota plays a significant role in detoxification. We analyzed the colon contents from three groups of pigs (fed either a standard diet, or a diet with 8 mg/kg DON or ZEN). Bacterial 16S rRNA gene amplicons were obtained from the colon contents, and sequenced using next-generation sequencing on the MiSeq platform. Overall, 2,444,635 gene sequences were generated, with ≥2000 sequences examined. Firmicutes and Bacteroidetes were the dominant phyla in all three groups. The sequences of

Topics: Animal Feed; Animals; Bacteria; Colon; Diet; Food Contamination; Microbiota; RNA, Ribosomal, 16S; Swine; Trichothecenes; Zearalenone

The aim of this study was to model fusarium mycotoxins against agronomic factors in order to identify those that have the greatest impact on mycotoxin levels in harvested wheat. To achieve this, fusarium mycotoxins levels were monitored, and associated agronomic data collected, in approximately 150 English wheat fields/year between 2006 and 2013. Results showed large seasonal variation in fusarium mycotoxin levels, with high levels in 2008 (13% and 29% exceeding legal limit for unprocessed soft wheat intended for human consumption for deoxynivalenol (DON) and zearalenone (ZON), respectively) and 2012 (10% and 15% exceeding legal limit for unprocessed soft wheat intended for human consumption for DON and ZON, respectively) and low levels in 2006 and 2011 (no samples exceeding legal limits for unprocessed soft wheat intended for human consumption for DON or ZON). Analysis of agronomic factors identified previous crop, cultivation and variety as the greatest risk factors. The greatest risk of mycotoxin development in grain was following maize as a previous crop and minimum tillage. The combined effect of these factors gave respective average DON and ZON levels 20 and 14 times higher than other previous crop and cultivation combinations. A newly quantified risk factor was harvest date. A 1-month delay in harvest resulted in a 10 and 25 times greater mean DON and ZON concentration, respectively, when compared to crops harvested around the long-term regional average harvest date. These results highlight the highly seasonal variation in fusarium mycotoxins in wheat and the agronomic factors that should be avoided to minimise fusarium mycotoxin levels in harvested wheat.

Topics: Crops, Agricultural; Fusarium; Humans; Trichothecenes; Triticum; Zearalenone

Zearalenone (ZEA) and deoxynivalenol (DON) are toxic fungal secondary metabolites, found mainly in contaminated food, that are associated with serious health problems. It is important to identify undesirable toxins and metabolites that may be present in human milk. The aim of this study was to evaluate human milk ZEA and DON levels, total daily intake of ZEA and DON; and their possible relationship with maternal dietary habits.. We enrolled 90 lactating mothers who had 7- to 90-day-old babies. A dietary questionnaire was completed by each of the mothers. Human milk samples were obtained from 90 mothers, and human milk ZEA and DON levels were evaluated with the solid-phase direct enzyme immunoassay. The total daily intake (TDI) was calculated for the 63 exclusively breastfed infants.. ZEA was detected in all human milk samples; median was 173.8 ng/L (35.7-682 ng/L). The calculated median TDI for ZEA was 33.0 ng/kg body weight (bw) (10.4-120.5 ng/kg) among exclusively breast-fed infants, none of them had a TDI that was above the previously defined threshold levels. Human milk ZEA levels were associated with the maternal consumption of meat, fish, dry fig, dried apricot, flaked red spice and spice. The median DON levels was 3924 ng/L (400-14997 ng/L). The median TDI of DON was 750 ng/kg (240-2774 ng/kg) among exclusively breastfed infants and 36% out of them, the TDI for DON was above the previously defined threshold level. Human milk DON levels were associated with the maternal meat consumption.. Our findings are indicative of dietary exposure to mycotoxins during the pregnancy and lactation periods in nursing mothers. Further, the excessive TDI values for DON observed in 36% of the exclusively breastfed infants point to the need for further regulations and recommendations on the dietary habits of pregnant/nursing mothers in order to avoid exposure to potential mycotoxins.

Topics: Adolescent; Adult; Diet; Dietary Exposure; Female; Humans; Meat; Milk, Human; Trichothecenes; Turkey; Young Adult; Zearalenone

Putative proton coupled di-peptide transporters, PTR2s, are found in filamentous fungi in different numbers and their function during fungal development and plant infection is unresolved. In Fusarium graminearum, the cause of head blight in cereals, we identified four putative PTR2 transporters (FgPTR2A-D). The genes did not cluster together in phylogenetic analyses and only FgPTR2A and FgPTR2C were able to complement a PTR2 deficient yeast mutant in uptake of di-peptides. All FgPTR2s are continuously expressed throughout the fungal lifecycle, although at different levels. In silico analyses of existing expression-data show that FgPTR2B is found at higher levels than the others in planta and during sexual development. Deletion mutants of FgPTR2A, FgPTR2C, and FgPTR2D had a higher production of deoxynivalenol (DON) and zearalenone and lower production of fusarielin H than the wild type. Perithecium development was reduced in these mutants but unaffected by deletion of FgPTR2B. Conidia production was reduced in the FgPTR2B mutant and unaffected by deletion of the other PTR2 transporters. Sexual development and secondary metabolite production are known to be linked at the regulatory level and the results suggest that PTR2s are active in nitrogen turnover and thereby influence signal processes.

Topics: Dipeptides; Fusarium; Gene Deletion; Gene Expression Profiling; Membrane Transport Proteins; Recombination, Genetic; Secondary Metabolism; Spores, Fungal; Trichothecenes; Zearalenone

Topics: Animal Feed; Animals; Endotoxemia; Escherichia coli; Food Contamination; Kinetics; Lipopolysaccharides; Swine; Trichothecenes; Zearalenone

Topics: Animals; Antioxidants; Apoptosis; Cell Survival; Cells, Cultured; Dose-Response Relationship, Drug; Drug Synergism; Lymphocytes; Oxidative Stress; Spleen; Swine; Trichothecenes; Zearalenone

The aim of this study was to investigate the occurrence of deoxynivalenol (DON) and zearalenone (ZEN) in unprocessed cereals and soybean sampled in 2014 and 2015 from different fields located in Croatian regions. A total of 306 samples were analysed for DON and 415 samples for ZEN concentrations using quantitative ELISA methods. In 2014, DON and ZEN were determined in all samples in the mean concentrations of 1,461 ± 2,265 µg/kg and 656 ± 853 µg/kg, respectively, while in 2015 these means were 2,687 ± 2,731 µg/kg and 1,140 ± 1,630 µg/kg, respectively. The cultivation year significantly (p < 0.05) influenced mycotoxin concentrations, whereas the influence of cultivation region was seen with ZEN for all cereals except soybean, and not seen with DON at all. A higher contamination determined during 2015 could be explained by high to extreme humidity evidenced in the period of cereals' growth and harvesting.

Topics: Croatia; Edible Grain; Food Contamination; Food Microbiology; Fusarium; Glycine max; Trichothecenes; Zearalenone

The detecting labels used for lateral flow immunoassays (LFAs) have been traditionally gold nanoparticles (GNPs) and, more recently, luminescent nanoparticles, such as quantum dots (QDs). However, these labels have low sensitivity and are costly, in particular, for trace detection of mycotoxins in cereals. Here, we provided a simple preparation procedure for amorphous carbon nanoparticles (ACNPs) and described multiplex LFAs employing ACNPs as labels (ACNP-LFAs) for detecting three Fusarium mycotoxins. The analytical performance of ACNPs in LFA was compared to GNPs and QDs using the same immunoreagents, except for the labels, allowing for their analytical characteristics to be objectively compared. The visual limit of detection for ACNP-LFAs in buffer was 8-fold better than GNPs and 2-fold better than QDs. Under optimized conditions, the quantitative limit of detection of ACNP-LFAs in maize was as low as 20 μg/kg for deoxynivalenol, 13 μg/kg for T-2 toxin, and 1 μg/kg for zearalenone. These measurements were much lower than the action level of these mycotoxins in maize. The accuracy and precision of the ACNP-LFAs were evaluated by analysis of spiked and incurred maize samples with recoveries of 84.6-109% and coefficients of variation below 13%. The results of ACNP-LFAs using naturally incurred maize samples showed good agreement with results from high-performance liquid chromatography-tandem mass spectrometry, indicating that ACNPs were more sensitive labels than and a promising alternative to GNPs used in LFAs for detecting mycotoxins in cereals.

Topics: Carbon; Food Contamination; Fusarium; Immunoassay; Mycotoxins; Nanoparticles; T-2 Toxin; Trichothecenes; Zea mays; Zearalenone

While numerous surveys highlighted the natural co-occurrence of mycotoxins in food, data about their toxicological combined effects is still limited. This is especially the case for chronic exposure conditions, although the latter are more representative of the mycotoxin risk associated with food consumption than acute exposure. In the present study, cell viability and gene expression levels of relevant hepatocyte-specific functions were evaluated for the HepaRG human liver cell line exposed to deoxynivalenol (DON) and/or zearalenone (ZEA) during 14, 28 and 42days at three subtoxic concentrations corresponding to i) the determined average exposure dose of French adult population, ii) the tolerable daily intake established by the Joint FAO/WHO Expert Committee and iii) the maximum level permitted by the European regulation in cereals intended for direct human consumption. For the latter, DON and DON+ZEA induced 90% cell mortality after 14days. In addition, depending on the considered toxin or mixture, doses and exposure periods, important variations of gene expression levels were observed. Despite the fact that in vitro conditions differ from the in vivo situation, the obtained results clearly highlighted that long-term toxicological effects of chronic exposure to mycotoxin combinations should be further investigated and, if necessary, taken into consideration at the regulatory level.

Topics: Cell Line; Cell Survival; Drug Administration Schedule; Drug Therapy, Combination; Gene Expression Regulation; Hepatocytes; Humans; Trichothecenes; Zearalenone

Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEA) are frequently occurring in feed of pigs together. The aim of this study was to evaluate the possible in vitro effects of DON and ZEA, alone or their combination on steroid secretion of porcine ovarian granulosa cells (GCs). A species-specific model with porcine ovarian GCs was used to study the potential endocrine disrupting effects of DON and ZEA alone and in co-exposure. Progesterone (P4) and estradiol (E2) were determined by radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA). The results of this study demonstrate that DON alone at the higher concentrations may act to stimulate P4 (at 1,000, 2,000, 3,000 and 5,000 ng mL

Topics: Animals; Cell Proliferation; Cells, Cultured; Endocrine Disruptors; Estradiol; Female; Fusarium; Granulosa Cells; Progesterone; Steroids; Swine; Trichothecenes; Zearalenone

While, Aflatoxin B

Topics: Aflatoxin B1; Animal Feed; Animals; Animals, Genetically Modified; Cell Line; Cell Survival; Larva; Liver; Trichothecenes; Water Pollutants, Chemical; Zearalenone; Zebrafish

Corn with naturally occurring aflatoxin (AF), wheat with naturally occurring doxynivalenol (DON), and barley with naturally occurring zearalenone (ZEA) were used to make rations for feeding turkey hen poults to 6 weeks of age. Control rations with equal amounts of corn, wheat, and barley were also fed. The control rations did contain some DON while both sets of rations contained ZEA. Within each grain source, there were 4 treatments: the control ration plus 3 rations each with a different feed additive which were evaluated for the potential to lessen potential mycotoxin effects on bird performance and physiology. The additives were Biomin BioFix (2 lb/ton), Kemin Kallsil (4 lb/ton), and Nutriad UNIKE (3 lb/ton). The mycotoxin rations reduced poult body weight (2.31 vs. 2.08 ± 0.02 kg) and increased (worsened) poult feed conversion (1.47 vs. 1.51 ± 0.01) at 6 wk. Feeding the poults the mycotoxin feed also resulted in organ and physiological changes typical of feeding dietary aflatoxin although a combined effect of AF, DON, and ZEA which cannot be dismissed. The feed additives resulted in improved feed conversion to 6 wk in both grain treatment groups. The observed physiological effect of feeding the additives was to reduce relative gizzard weight for both groups and to lessen the increase in relative kidney weight for the birds fed the mycotoxin feed. In conclusion, the feed additives used in this study did alleviate the effect of dietary mycotoxins to some degree, especially with respect to feed conversion. Further studies of longer duration are warranted.

Topics: Aflatoxins; Animal Feed; Animals; Diet; Dietary Supplements; Female; Mycotoxins; Protective Agents; Random Allocation; Trichothecenes; Turkeys; Zearalenone

(1) Background and (2) Methods: A 14-day in vivo, multitoxic (pure mycotoxins) rat experiment was conducted with zearalenone (ZEA; 15 μg/animal/day), deoxynivalenol (DON; 30 μg/animal/day) and fumonisin B₁ (FB₁; 150 μg/animal/day), as individual mycotoxins, binary (FD, FZ and DZ) and ternary combinations (FDZ), via gavage in 1 mL water boluses. (3) Results: Body weight was unaffected, while liver (ZEA↑ vs. DON) and kidney weight (ZEA↑ vs. FDZ) increased. Hepatocellular membrane lipid fatty acids (FAs) referred to ceramide synthesis disturbance (C20:0, C22:0), and decreased unsaturation (C22:5 n3 and unsat. index), mainly induced by DON and to a lesser extent by ZEA. The DON-FB₁ interaction was additive on C20:0 in liver lipids. In renal phospholipids, ZEA had the strongest effect on the FA profile, affecting the saturated (C18:0) and many n6 FAs; ZEA was in an antagonistic relationship with FB₁ (C18:0) or DON (C18:2 n6, C20:1 n9). Hepatic oxidative stress was the most expressed in FD (reduced glutathione and glutathione peroxidase), while the nephrotoxic effect was further supported by lipid peroxidation (malondialdehyde) in the DON treatment. (4) Conclusions: In vivo study results refer to multiple mycotoxin interactions on membrane FAs, antioxidants and lipid peroxidation compounds, needing further testing.

Topics: Animals; Fatty Acids; Fumonisins; Kidney; Liver; Male; Membrane Lipids; Organ Size; Rats, Wistar; Trichothecenes; Zearalenone

Topics: Edible Grain; Fusarium; Plant Diseases; Trichothecenes; Triticum; Zearalenone

Topics: Chromosome Mapping; Crosses, Genetic; Disease Resistance; Genetic Linkage; Genotype; Gibberella; Phenotype; Plant Breeding; Plant Diseases; Polymorphism, Single Nucleotide; Quantitative Trait Loci; Trichothecenes; Zea mays; Zearalenone

Mycotoxins are toxic secondary metabolites produced by a range of fungi and are common contaminants of agricultural crops. These toxins are chemically diverse and structurally stable, enabling them to enter the food chain which can lead to numerous adverse health effects in animals and humans. Although mycotoxin exposure is associated with the development of several cancers, it has proved challenging to show a direct connection between exposure and oncogenic change. This study investigates the in vitro cytotoxicity, molecular mechanisms and secondary signalling responses associated with the exposure to three major mycotoxins, fumonisin B1 (FB1), deoxynivalenol (Don) and zearalenone (Zea). The cytotoxicity of FB1, Don and Zea were investigated in cultured HepG2 and Caco-2 cells using cell viability assays as well as flow cytometry. FB1 proved to be less cytotoxic than its counterparts, while Don and Zea demonstrated high cytotoxicity through an apoptotic mechanism. Expression profiles of 84 genes involved in mediating communication between tumour cells and the cellular mediators of inflammation as well as the innate immune system were also studied. The expression profiles associated with the different mycotoxins were further explored for functional networks, biological functions, canonical pathways, toxicological association as well as to predict network associations between the differentially expressed genes. RT-qPCR revealed the significant differential expression of 46 genes, including the expression of several genes strongly associated with cancer and aberrant inflammatory signalling, after mycotoxin exposure. Aberrant inflammatory signalling seems to be a credible contributing factor that initiates the malignant change observed in cells exposed to mycotoxins.

Topics: Apoptosis; Caco-2 Cells; Caspase 3; Caspase 7; Computer Simulation; Flow Cytometry; Fumonisins; Gene Expression Profiling; Gene Expression Regulation; Hep G2 Cells; Humans; L-Lactate Dehydrogenase; Mycotoxins; Signal Transduction; Trichothecenes; Zearalenone

Dairy cows experience a negative energy balance at the onset of lactation which results in an enhanced vulnerability for infectious diseases. Any dietary imbalances, including Fusarium toxin contamination, might therefore exacerbate this situation. The aim of the present investigations was to study the effects of increasing dietary concentrations of deoxynivalenol (DON) and zearalenone (ZEN) on clinical-chemical, haematological and immunological traits up to week 14 of lactation. For this purpose, ten cows each were assigned to a control group (CON; 0.02 mg ZEN and 0.06 mg DON per kg diet at 88 % DM), toxin level 1 (TOX-1; 0.29 mg ZEN and 2.31 mg DON per kg diet at 88 % DM) and toxin level 2 (TOX-2; 0.58 mg ZEN and 4.61 mg DON per kg diet at 88 % DM). The measured values of most parameters were affected by parturition but only a few of them were further modified by dietary treatment. For example, the time-dependent decrease in haemoglobin concentration, haematocrit and erythrocyte counts occurred at a significantly higher level for group TOX-2 while a serum glucose increase was missing in this group. Proportions of CD4+ and CD8+ cells decreased significantly over time solely in group TOX-2 while the CD4+/CD8+ ratio remained uninfluenced. Ability of granulocytes to mount an oxidative burst tended to increase at the end of the study in groups TOX-1 and TOX-2 while the opposite was observed in group CON. The results of this time-limited study indicate that feeding of Fusarium-toxin contaminated diets in early lactation affects health related parameters without compromising milking performance. However, long-term consequences of the observed effects on health need to be addressed in further studies.

Topics: Animal Feed; Animals; Blood Chemical Analysis; Blood Glucose; Cattle; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Diet; Hemoglobins; Lactation; Lymphocyte Count; Poisoning; Trichothecenes; Zearalenone

The aim of this study was to determine whether exposure to low doses of ZEN + DON induces changes in serum biochemical and hematological parameters in pre-pubertal gilts. In the evaluated groups, minor but statistically significant changes were noted in selected serum biochemical parameters, including glucose, total cholesterol, ALT, AST, AP, total bilirubin, P

Topics: Animals; Blood Glucose; Body Weight; Cholesterol; Erythrocyte Count; Glucose; Leukocyte Count; Sexual Maturation; Swine; Trichothecenes; Zearalenone

Topics: Antibodies, Immobilized; Edible Grain; Food Contamination; Gold Colloid; Metal Nanoparticles; Mycotoxins; Quantum Dots; Trichothecenes; Zearalenone

Topics: Antibodies, Monoclonal; Cadmium; Drug Compounding; Humans; Immunoassay; Immunoconjugates; Indium; Limit of Detection; Mycotoxins; Nanostructures; Phosphines; Quantum Dots; Silicon Dioxide; Solubility; Sulfides; Trichothecenes; Triticum; Water; Zea mays; Zearalenone; Zinc Compounds

Fusarium mycotoxins deoxynivalenol (DON) and zearalenone (ZEN) are the mostcommon contaminants in cereals worldwide, causing a wide range of adverse health effects onanimals and humans. Many environmental factors can affect the production of these mycotoxins.Here, we have used response surface methodology (RSM) to optimize the Fusarium graminearumstrain 29 culture conditions for maximal toxin production. Three factors, medium pH, incubationtemperature and time, were optimized using a Box-Behnken design (BBD). The optimizedconditions for DON production were pH 4.91 and an incubation temperature of 23.75 °C for 28 days,while maximal ZEN production required pH 9.00 and an incubation temperature of 15.05 °C for 28days. The maximum levels of DON and ZEN production were 2811.17 ng/mL and 23789.70 ng/mL,respectively. Considering the total level of DON and ZEN, desirable yields of the mycotoxins werestill obtained with medium pH of 6.86, an incubation temperature of 17.76 °C and a time of 28 days.The corresponding experimental values, from the validation experiments, fitted well with thesepredictions. This suggests that RSM could be used to optimize Fusarium mycotoxin levels, whichare further purified for use as potential mycotoxin standards. Furthermore, it shows that acidic pHis a determinant for DON production, while an alkaline environment and lower temperature(approximately 15 °C) are favorable for ZEN accumulation. After extraction, separation andpurification processes, the isolated mycotoxins were obtained through a simple purification process,with desirable yields, and acceptable purity. The mycotoxins could be used as potential analyticalstandards or chemical reagents for routine analysis.

Topics: Fusarium; Trichothecenes; Zea mays; Zearalenone

This study investigated the fungal diversity and presence of deoxynivalenol and zearalenone in 150 samples of freshly harvested wheat grains collected in three regions of Brazil (Sao Paulo, Parana, and Rio Grande do Sul). Analysis of the mycobiota showed a predominance of Alternaria sp., Fusarium sp. and Epicoccum sp. Microdochium nivale (23%), a fungus rarely found in Brazilian crops, was detected in Sao Paulo. Four members of the Fusarium graminearum species complex were isolated: F. graminearum s.s. (37%), Fusarium meridionale (46%), Fusarium cortaderiae (13%), and Fusarium austroamericanum (3%). Toxin analysis revealed 99% contamination with deoxynivalenol (mean 706 μg/kg). The frequency of zearalenone varied greatly across regions: wheat grains from Rio Grande do Sul (84%) and Sao Paulo (12%) had median concentrations of 70.9 and 57.9 μg/kg, respectively. ZEA was not detected in the samples from Parana. A total of six samples were above the maximum tolerated level recommended by the European Commission for ZEA in wheat grains. This study provided new insights into the natural mycobiota of Brazilian wheat, demonstrating contamination of most samples with deoxynivalenol and high frequency of zearalenone in samples from Rio Grande do Sul.

Topics: Brazil; Chromatography, Liquid; Crops, Agricultural; DNA, Fungal; Food Contamination; Food Microbiology; Fusarium; Limit of Detection; Reproducibility of Results; Tandem Mass Spectrometry; Trichothecenes; Triticum; Zearalenone

Diet change and fatness are supposed to challenge the immune system of the cow. Therefore, immunological and haematological consequences of adaptation to and continued feeding of a high-energy diet were studied in eight non-pregnant, non-lactating Holstein cows over 16 weeks. Blood haptoglobin concentration remained unaltered, suggesting that an acute phase reaction was not induced. Stimulation ability of peripheral blood mononuclear cells and stimulated oxidative burst capacity of granulocytes increased significantly in the course of the experiment after an initial drop. While total leucocyte counts increased, the proportion of granulocytes increased and that of lymphocytes decreased at the same time as the ratio of CD4(+)/CD8(+) lymphocytes did. Capability of rumen microbes to detoxify the immune-modulating mycotoxin deoxynivalenol (DON) was not compromised as indicated by the exclusive presence of de-DON as the detoxified DON metabolite in blood. In conclusion, both diet change and prolonged positive energy balance influenced the bovine immune system.

Topics: Adipose Tissue; Animal Feed; Animals; Cattle; Dairying; Diet; Energy Metabolism; Female; Leukocytes, Mononuclear; Mycotoxins; Respiratory Burst; Trichothecenes; Zearalenone

The effects of deoxynivalenol (DON) and zearalenone (ZEA) on reproduction in ruminants are unclear. This study was performed to evaluate the impact of DON and ZEA hydroxylated metabolites, α-zearalenol (α-Zol) and β-zearalenone (β-Zol), on cell proliferation, steroidogenesis and gene expression using bovine granulosa cells (GC). Cell proliferation was negatively affected after exposure to β-Zol at 31 μM and after exposure to α-Zol (3.1 μM) alone and combined with DON (3.3 μM). DON and α-Zol decreased steroidogenesis, while β-Zol at high concentration had stimulatory effects. DON and β-Zol increased CYP19A1 mRNA abundance. CYP11A1 mRNA abundance was stimulated by DON, alone and combined with α-Zol and β-Zol, whereas was inhibited by β-Zol alone. Generally mycotoxins effects on cell proliferation, steroidogenesis and gene expression were influenced by the presence or absence of IGF1. In conclusion DON and ZEA metabolites may impair in vitro cell proliferation, steroid production and gene expression in cattle.

Topics: Animals; Cattle; Cell Proliferation; Dose-Response Relationship, Drug; Female; Gene Expression; Granulosa Cells; In Vitro Techniques; Steroids; Trichothecenes; Zearalenone

This study aimed to evaluate the effects of the Fusarium toxin zearalenone (ZEA) and deoxynivalenol (DON) on splenic antioxidant functions, IFN levels, and T-cell subsets in mice. Herein, 360 mice were assigned to nine groups for a 12-day study. Mice were administered an intraperitoneal injection for 4 consecutive days with different concentrations of ZEA alone, DON alone, or ZEA+DON. Spleen and blood samples were collected on days 0, 3, 5, 8, and 12. Mice in each of the experimental groups showed dysreglated splenic antioxidant functions, IFN levels, and T-cell subset frequencies, suggesting that the immune system had been affected. The ZEA+DON-treated groups, especially the group that received a higher concentration of ZEA+DON (Group D2Z2), showed more obvious effects on the dysregulation of splenic antioxidant functions, IFN levels, and T-cell subsets. This finding suggested that DON and ZEA exerted synergistic effects.

Topics: Animals; Drug Synergism; Fusarium; Injections, Intraperitoneal; Interferons; Malondialdehyde; Mice; Mycotoxins; Spleen; T-Lymphocyte Subsets; Trichothecenes; Zearalenone

Between 2012 and 2014, 2528 feed ingredient and complete feed samples were collected from central China. Numbers of 2083, 255 and 190 samples were analysed for aflatoxin B1 (AFB1), zearalenone (ZEN) and deoxynivalenol (DON), respectively, by high-performance liquid chromatography in combination with UV or fluorescence detection. The incidence rates of AFB1, ZEN and DON contamination of feed ingredients and complete feeds were 33.9%, 90.2% and 77.4%, respectively. The percentage of positive samples for AFB1 ranged from 13.1% to 97.1%. Cottonseed meal presented the most serious contamination by AFB1. ZEN and DON contamination levels of feeds ranged from 50% to 100%, indicating serious contamination over the studied 3-year period. This study demonstrates that AFB1, ZEN and DON contamination of feeds in central China is serious and differs over the years. Feeds are mostly contaminated with ZEN, followed by DON and AFB1.

Topics: Aflatoxin B1; Animal Feed; Animals; China; Chromatography, High Pressure Liquid; Diet; Food Contamination; Gossypium; Humans; Seeds; Trichothecenes; Zearalenone

The worldwide contamination of grains designated to human and animal feeding with Fusarium mycotoxins is a significant problem. Among Fusarium mycotoxins, deoxynivalenol (DON) and zearalenone (ZEA) are the most prevalent mycotoxins found in cereals. Co-occurrence of DON and ZEA is also very frequent and indicates that these mycotoxins might be involved in a wide range of synergistic or additive interactions. Both mycotoxins have been linked to various male reproduction problems including downregulation of steroidogenesis. In this study, the impact of DON and ZEA alone or in combination on the viability and steroid production of Leydig cell line MA-10 was determined. The ability of vitamin E, sesamin and their combination to prevent oxidative stress and restore progesterone secretion in DON- and ZEA-exposed cells was also determined. Results showed that MA-10 cells were more sensitive to the effect of DON compared to ZEA. DON and ZEA also significantly reduced MA-10 progesterone secretion after forskolin activation but no significant interactions between DON and ZEA were detected. Preventive treatment with the combination of vitamin E and sesamin significantly reduced ROS production and increased cell survival after exposition to DON and ZEA. However this treatment failed to restore normal progesterone secretion. In conclusion, both DON and ZEA are deleterious to steroidogenesis in Leydig cells. Prevention of oxidative stress caused by DON and ZEA was effective to restore cell viability but failed to restore other functions of Leydig cells suggesting that ROS production is not the main cause of steroidogenic failure in DON and ZEA treated MA-10 cells.

Topics: Animals; Antioxidants; Cell Line; Cell Survival; Colforsin; Gene Expression Regulation; Leydig Cells; Male; Mice; Mycotoxins; Oxidative Stress; Progesterone; Reactive Oxygen Species; Steroids; Trichothecenes; Zearalenone

A survey was conducted to determine whether mycotoxins were present in the foods consumed by red-crowned cranes (Grus japonensis) in the Yancheng Biosphere Reserve, China. Collected in the reserve's core, buffer, and experimental zones during overwintering periods of 2013 to 2015, a total of 113 food samples were analyzed for aflatoxin B1, deoxynivalenol, zearalenone, T-2 toxin, and ochratoxin A using high performance liquid chromatography (HPLC). The contamination incidences vary among different zones and the mycotoxins levels of different food samples also presented disparity. Average mycotoxin concentration from rice grain was greater than that from other food types. Among mycotoxin-positive samples, 59.3% were simultaneously contaminated with more than one toxin. This study demonstrated for the first time that red-crowned cranes were exposed to mycotoxins in the Yancheng Biosphere Reserve and suggested that artificial wetlands could not be considered good habitats for the birds in this reserve, especially rice fields.

Topics: Aflatoxin B1; Animals; Birds; Endangered Species; Environmental Exposure; Food Analysis; Food Contamination; Mycotoxins; Ochratoxins; T-2 Toxin; Trichothecenes; Zearalenone

The Fusarium graminearum species complex infects several cereals and causes the reduction of grain yield and quality. Many factors influence the extent of Fusarium infection and mycotoxin levels. Such factors include crop rotation. In the present study, we explored the effect of rice or maize as former crops on mycotoxin accumulation in wheat grains.. More than 97% of samples were contaminated with deoxynivalenol (DON). DON concentrations in wheat grains from rice and maize rotation fields were 884.37 and 235.78 µg kg(-1) . Zearalenone (ZEN) was detected in 45% of samples which were mainly collected from maize-wheat rotation systems. Fusarium strains were isolated and more F. graminearum sensu stricto (s. str.) isolates were cultured from wheat samples obtained from maize rotation fields. DON levels produced by Fusarium isolates from rice rotation fields were higher than those of samples from maize rotation fields.. Rice-wheat rotation favours DON accumulation, while more ZEN contamination may occur in maize-wheat rotation models. Appropriate crop rotation may help to reduce toxin levels in wheat grains. © 2016 Society of Chemical Industry.

Topics: China; Crop Production; Crops, Agricultural; Environmental Pollutants; Food Contamination; Fusarium; Molecular Typing; Mycological Typing Techniques; Mycotoxins; Oryza; Seeds; Soil Microbiology; Spatio-Temporal Analysis; Trichothecenes; Triticum; Zea mays; Zearalenone

Fusarium mycotoxins deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN) were investigated in wheat from the 2009 and 2010 crop years. Samples (n = 745) from commercial fields were collected in four wheat producing regions (WPR) which differed in weather conditions. Analyses were performed using HPLC-DAD. Contamination with ZEN, DON and NIV occurred in 56, 86 and 50%, respectively. Also, mean concentrations were different: DON = 1046 µg kg(-1), NIV < 100 µg kg(-1) and ZEN = 82 µg kg(-1). Co-occurrence of ZEN, DON and NIV was observed in 74% of the samples from 2009 and in 12% from 2010. Wet/cold region WPR I had the highest mycotoxin concentration. Wet/moderately hot region WPR II had the lowest mycotoxin levels. Furthermore, the mean concentration of each mycotoxin was higher in samples from 2009 as compared with those from 2010. Precipitation during flowering or harvest periods may explain these results.

Topics: Brazil; Chromatography, High Pressure Liquid; Climate; Diet; Edible Grain; Food Contamination; Fusarium; Humans; Trichothecenes; Triticum; Weather; Zearalenone

Four diets contaminated with 1.1 to 5.0 mg/kg deoxynivalenol (DON) and 0.4 to 2.4 mg/kg zearalenone (ZEA) were fed to four groups of six growing Large White pigs. Urine samples were collected after 3 to 4 days and again after 6 to 7 days on the diets. On each sampling day, half of the animals were sampled in the morning, after an 8-h fast, and the other half were sampled in the afternoon, after 7 h of ad libitum access to feed. The urinary concentrations of DON, DON-glucuronide, DON-3-sulphate, de-epoxy-DON, as well as of ZEA, ZEA-14-glucuronide, α-zearalenol and α-zearalenol-14-glucuronide, analysed using LC-MS/MS, were used to calculate urinary DON and ZEA equivalent concentrations (DONe and ZEAe). The urinary concentration of DONe (P < 0.001), but not of ZEAe (P = 0.31), was lower in the fasted than that in the fed animals. The urinary DONe/creatinine and ZEAe/creatinine ratios were highly correlated with DON and ZEA intake per kg body weight the day preceding sampling (r = 0.76 and 0.77; P < 0.001). The correlations between DON intake during the 7 h preceding urine sampling in the afternoon and urinary DONe/creatinine ratio (r = 0.88) as well as between mean ZEA intake during 3 days preceding urine sampling and urinary ZEAe/creatinine ratio (r = 0.84) were even higher, reflecting the plasma elimination half-time of several hours for DON and of more than 3 days for ZEA. ZEAe analysed in enzymatically hydrolysed urine using an ELISA kit was highly correlated with the LC-MS/MS data (r = 0.94). The urinary DONe and ZEAe to creatinine ratios, analysed in pooled urine samples of several pigs fed the same diet, can be used to estimate their exposure to DON and ZEA.

Topics: Animal Feed; Animals; Biomarkers; Fungi; Mycotoxins; Swine; Trichothecenes; Zea mays; Zearalenone

The Fusarium mycotoxins deoxynivalenol (DON), zearalenone (ZEN) and T-2 frequently contaminate grain crops in Middle and Eastern Europe. In this survey, 116 cereal samples (maize, wheat, barley and oat) were examined for DON, ZEN and T-2 mycotoxins. Samples were collected from different areas in two Hungarian regions (North and South Transdanubia). The method of analysis was indirect competitive ELISA. Maize was the most contaminated grain regarding DON (86%), ZEN (41%) and T-2 (55%) toxins. The average results of the deoxynivalenol and zearalenone tests of maize proved to be significantly higher than those of barley or oat. DON was the most represented Fusarium mycotoxin followed by T-2 and ZEN. The examination of these mycotoxins would be necessary at a larger scale as to re-evaluate permissible levels, so increase of the monitoring programme would be advisable for the future.

Topics: Avena; Crops, Agricultural; Diet; Edible Grain; Food Contamination; Food Microbiology; Fusarium; Hordeum; Humans; Hungary; Mycotoxins; T-2 Toxin; Trichothecenes; Triticum; Zea mays; Zearalenone

Seventy-two piglets (6.0 kg BW) were randomly distributed within six different dietary treatments to evaluate the effect of deoxynivalenol (DON) and the potential of four antioxidant feed additives in mitigating the adverse effects of DON on growth performances and oxidative status. Dietary treatments were as follows: control diet 0.8 mg/kg DON; contaminated diet (DON-contaminated diet) 3.1 mg/kg DON; and four contaminated diets, each supplemented with a different antioxidant feed additive, DON + vitamins, DON + organic selenium (Se)/glutathione (GSH), DON + quercetin, and DON + COMB (vitamins + Se/GSH + quercetin from the other treatments). Although DON was the main mycotoxin in the contaminated diet, this diet also contained 1.8 mg/kg of zearalenone (ZEN). The "mycotoxin" effects therefore included the combined effect of these two mycotoxins, DON, and ZEN. The DON-ZEN ingestion did not affect growth performances, average daily gain (ADG), average daily feed intake (ADFI), and feed efficiency (G:F ratio), but partially induced oxidative stress in weaned pigs as shown by increased malondialdehyde (MDA) content in the plasma and superoxide dismutase (SOD) activity in liver (P < 0.05). However, no change in the activity of other antioxidant enzymes or GSH concentrations was observed in plasma and liver of piglets fed the DON-contaminated diet (P > 0.05). Supplementation with individual antioxidant feed additive had a limited effect in weaned pigs fed DON-ZEN-contaminated diets. Combination of antioxidants (vitamins A, C, and E, quercetin, and organic Se/GSH) reduced plasma and liver MDA content and SOD activity in liver (P < 0.05) of piglets fed DON-ZEN-contaminated diets. Furthermore, this combination also reduced MDA content in the ileum (P < 0.05), although activity of glutathione peroxidases (GPx), SOD or catalase (CAT) in the ileum was not affected by DON-ZEN contamination or antioxidant supplements. In conclusion, DON-ZEN contamination induced oxidative stress in weaned pigs and combination of antioxidant feed additives restored partially the oxidative status. Further studies will be necessary to assess whether the effects of antioxidant feed additives on oxidative status are specific when feed is contaminated with DON-ZEN.

Topics: Animal Feed; Animals; Antioxidants; Female; Food Additives; Fusarium; Intestinal Mucosa; Liver; Male; Mycotoxins; Swine; Trichothecenes; Weaning; Zea mays; Zearalenone

Fusarium graminearum is a destructive pathogen of cereals that can cause stalk rot in maize. Stalk rot results in yield losses due to impaired grain filling, premature senescence, and lodging, which limits production and harvesting of ears. In addition, mycotoxins can make infected tissues unfit for silage. Our objectives were to evaluate the natural variation in stalk rot resistance among maize inbreds, to establish whether deoxynivalenol (DON)- and zearalenone (ZEA)-deficient strains are pathogenic on a panel of diverse inbreds, and to quantify the accumulation of DON in infected stalk tissue. Wild-type F. graminearum and mycotoxin mutants (DON and ZEA) were used to separately inoculate stalks of 9-week-old plants of 20 inbreds in the greenhouse. Plants were evaluated for lesion area at the inoculation point at 0, 2, 14, and 28 days postinoculation and tissues around lesions were sampled to determine the DON content. Regardless of their ability to produce DON or ZEA, all tested F. graminearum strains caused stalk rot; however, significant differences in disease levels were detected. Among the tested inbreds, Mp717 was resistant to all three F. graminearum strains while Mp317 and HP301 were only partially resistant. Accumulation of DON was significantly lower in infected stalks of the resistant and partially resistant inbreds than the susceptible inbreds. Analysis of the 20 inbreds using data from 17 simple-sequence repeats revealed population structure among the individuals; however, there was no association between genetic clustering and stalk rot resistance. These findings are an additional step toward breeding maize inbreds suitable for planting in fields infested with F. graminearum.

Topics: DNA, Plant; Fusarium; Gene Expression Regulation, Fungal; Genetic Predisposition to Disease; Mutation; Plant Diseases; Trichothecenes; Zea mays; Zearalenone

The present feeding study was carried out to examine the effects of Fusarium toxin-contaminated diets on performance and slaughtering characteristics and on the transfer of the Fusarium toxins zearalenone (ZEN), deoxynivalenol (DON) and their metabolites into physiological matrices. A total of 61 bulls (483 ± 46 kg) were fed with graded proportions of Fusarium toxin-contaminated feed over a period of 10 weeks. The total mixed rations (TMR) consisted of 47 % grass silage, 20 % press pulp silage, and 33 % concentrate on dry matter (DM) basis. Increasing toxin concentrations were achieved by the exchange of control maize with Fusarium toxin-contaminated maize in the concentrates. Thus, dietary toxin concentrations between 0.08 and 0.69 mg ZEN and 0.36 and 8.31 mg DON per kg DM were covered by the four feeding groups. Based on increasing DM intake with increasing mycotoxin contaminations of the diet, the live weight gain and energy intake differed significantly between the groups. No effects were observed on slaughtering characteristics and organ weights. ZEN, α-zeralenol, β-zeralenol (β-ZEL), zeralanone, α-zearalanol, β-zearalanol, DON, and de-deepoxy-DON (de-DON) were simultaneously determined in urine, plasma, and liquor whereby quantifiable concentrations of ZEN, β-ZEL, DON, and de-DON were found in urine, of DON and de-DON in plasma, and solely of de-DON in liquor. Based on overall results it can be concluded that current EU-guidance values for critical concentrations of DON and ZEN can be regarded as safe levels also for growing bulls. Urine and blood toxin residue levels can be used to assess exposure of bulls.

Topics: Animal Feed; Animal Structures; Animals; Body Weight; Cattle; Cattle Diseases; Cerebrospinal Fluid; Food Contamination; Male; Plasma; Poisoning; Trichothecenes; Urine; Zea mays; Zearalenone

Zearalenone and deoxynivalenol are secondary metabolites of fungi of the genus Fusarium. The presence of mycotoxins in cereals and the resulting contamination of feeds and foods pose health risks for animals and humans. The dangers associated with high doses of mycotoxins have been extensively researched but very little is known about NOAEL (No Observed Adverse Effect Level) doses or exposure to a combination of mycotoxins (mixed mycotoxicoses). The aim of this study was to determine the effects of six-week exposure to NOAEL doses of individual and combined mycotoxins on the subpopulations of CD4⁺8(-), CD4(-)8⁺ and CD4⁺8⁺ lymphocytes in the peripheral blood of pigs. The experiment was performed on 72 gilts with average body weight of 25 kg, divided into three experimental groups (E1, E2 and E3, administered zearalenone (ZEN), deoxynivalenol (DON) and ZEN + DON, respectively, on a daily basis) and a control group (C) receiving placebo. Changes in lymphocyte subpopulations were evaluated by flow cytometry at weekly intervals (experimental days 7, 14, 21, 28, 35 and 42). A linear increase in the percentage of CD4⁺8⁺ lymphocytes was highly correlated with time (r = 0.682) in group C. The correlations and linear increase in the above subpopulation were disrupted in the remaining groups. In group E3, a statistically significant (p < 0.05) decrease in CD4⁺8⁺ counts was observed in week 5, which could point to a transient depletion of regulatory mechanisms of immune responses. The noted results also suggest that in mixed mycotoxicosis, ZEN and DON exerted stronger immunomodulatory effects.

Topics: Animals; Food Contamination; Fusarium; Lymphocyte Subsets; Mycotoxins; No-Observed-Adverse-Effect Level; Swine; Trichothecenes; Zearalenone

Dietary exposure of Hong Kong adults to mycotoxins and their metabolites including aflatoxins (AFs), ochratoxin A (OTA), fumonisins (FNs), deoxynivalenol (DON), acetyldeoxynivalenols (AcDONs) and zearalenone (ZEA) was estimated using the Total Diet Study (TDS) approach to assess the associated health risk to the local people. Sixty commonly consumed food items, collected in four seasons, were sampled and prepared as consumed. These mycotoxins were primarily found at low levels. The highest mean levels (upper bound) were: AFs, 1.50 µg kg(-)(1) in legumes, nuts and seed; OTA, 0.22 µg kg(-)(1) in sugars and confectionery; FNs, 9.76 µg kg(-)(1) in cereals and their products; DON and AcDONs, 33.1 µg kg(-)(1) in cereals and their products; and ZEA, 53.8 µg kg(-)(1) in fats and oils. The estimated dietary exposures of Hong Kong adults to the mycotoxins analysed were well below the respective health-based guidance values, where available. For AFs, the upper-bound exposure for high consumers is 0.0049 µg kg bw(-)(1) day(-)(1), which was estimated to contribute to about 7.7 (< 1%) of liver cancer cases when compared with 1222 liver cancer cases per year in Hong Kong. The percentage contributions of the estimated 95th percentile dietary exposures (lower and upper bound) to the health-based guidance values of individual mycotoxins were: ochratoxin A, 3.6-9.2%; fumonisins, 0.04-8.5%; deoxynivalenol and acetyldeoxynivalenols, 21.7-28.2%; and zearalenone 3.3-34.5%. The findings indicate that dietary exposures to all the mycotoxins analysed in this study were unlikely to pose an unacceptable health risk to the Hong Kong population.

Topics: Adult; Aflatoxins; Diet; Environmental Exposure; Food Contamination; Fumonisins; Hong Kong; Humans; Liver Neoplasms; Maximum Allowable Concentration; Mycotoxins; Nutrition Surveys; Ochratoxins; Seasons; Surveys and Questionnaires; Trichothecenes; Zearalenone

Over a 4-year period (2010-13), a survey aiming at determining the occurrence of Fusarium spp. and their relations to mycotoxins in mature grains took place in southern Belgium. The most prevalent species were F. graminearum, F. avenaceum, F. poae and F. culmorum, with large variations between years and locations. An even proportion of mating type found for F. avenaceum, F. culmorum, F. cerealis and F. tricinctum is usually a sign of ongoing sexual recombination. In contrast, an unbalanced proportion of mating type was found for F. poae and no MAT1-2 allele was present in the F. langsethiae population. Genetic chemotyping indicates a majority of deoxynivalenol (DON)-producing strains in F. culmorum (78%, all 3-ADON producers) and F. graminearum (95%, mostly 15-ADON producers), while all F. cerealis strains belong to the nivalenol (NIV) chemotype. Between 2011 and 2013, DON, NIV, enniatins (ENNs) and moniliformin (MON) were found in each field in various concentrations. By comparison, beauvericin (BEA) was scarcely detected and T-2 toxin, zearalenone and α- and β-zearalenols were never detected. Principal component analysis revealed correlations of DON with F. graminearum, ENNs and MON with F. avenaceum and NIV with F. culmorum, F. cerealis and F. poae. BEA was associated with the presence of F. tricinctum and, to a lesser extent, with the presence of F. poae. The use of genetic chemotype data revealed that DON concentrations were mostly influenced by DON-producing strains of F. graminearum and F. culmorum, whereas the concentrations of NIV were influenced by the number of NIV-producing strains of both species added to the number of F. cerealis and F. poae strains. This study emphasises the need to pay attention to less-studied Fusarium spp. for future Fusarium head blight management strategies, as they commonly co-occur in the field and are associated with a broad spectrum of mycotoxins.

Topics: Belgium; Depsipeptides; DNA, Fungal; Edible Grain; Food Contamination; Fusarium; Genes, Mating Type, Fungal; Humans; Mycotoxins; Principal Component Analysis; Trichothecenes; Zearalenone

The levels of 26 mycotoxins were determined in 147 samples of the grain of cereals cultivated in five regions of Poland during the 2014 growing season. The HPLC-HRMS (time-of-flight) analytical technique was used. An analytical procedure to simultaneously determine 26 mycotoxins in grain was developed, tested and verified. Samples from eastern and southern Poland were more contaminated with mycotoxins than the samples from northern and western Poland. Toxins produced by Fusarium fungi were the main contaminants found. Some deoxynivalenol (DON) was found in 100% of the tested samples of wheat (Osiny, Borusowa, Werbkowice), triticale, winter barley and oats, while the maximum permissible DON level (as defined in the EU Commission Regulation No. 1881/2006) was exceeded in 10 samples. Zearalenone (ZEN), DON metabolites and enniatins were also commonly found. The presence of mycotoxins in grain reflected the prevailing weather conditions during the plant flowering/earing stages, which were favorable for the development of blight. Among all investigated wheat genotypes, cv. Fidelius was the least contaminated, while Bamberka, Forkida and Kampana were the most contaminated. However, the single-factor ANOVA analysis of variance did not reveal (at a statistical significance level α = 0.05) any differences between levels of mycotoxins in individual genotypes. Triticale was the most contaminated grain among all of the tested varieties. ZEN, DON and the sum of 3-acetyldexynivalenol and 15-acetyldeoxynivalenol (3- and 15-ADON) were found in 100% of the tested triticale samples at concentrations within the 4-86, 196-1326 and 36-374 µg·kg(-1) range, respectively. Of particular concern was the fact that some "emerging mycotoxins" (enniatins) (in addition to commonly-known and legally-regulated mycotoxins) were also found in the tested triticale samples (enniatin B (Enn-B), enniatin B1 (Enn-B1), enniatin A-1 (Enn-A1), 100% of samples, and enniatin A (Enn-A), 70% of samples). Depending on the toxin, they were found at levels between 8 and 3328 µg·kg(-1).

Topics: Avena; Edible Grain; Food Contamination; Fusarium; Genotype; Hordeum; Mycotoxins; Poland; Trichothecenes; Triticale; Triticum; Weather; Zearalenone

Both deoxynivalenol (DON), zearalenone (ZEN), and their metabolites are known to modulate immune cells in various species whereby viability and proliferation are influenced. Such effects were rarely examined in horses. Therefore, one aim of the present study was to titrate the inhibitory concentrations of DON, 3-acetyl-DON (3AcDON), de-epoxy-DON (DOM-1), ZEN, and α- and β-zearalenol (ZEL) at which viability and proliferation of equine PBMC were reduced by 50 % (IC50) and 10 % (IC10) in vitro. For evaluation of practical relevance of the in vitro findings, a further aim was to screen horses for the background occurrence of DON, ZEN, and their metabolites in systemic circulation and to relate toxin residues both to the inhibitory toxin concentrations and to hematological and clinical-chemical characteristics.The IC50 (μM) for DON, 3AcDON, β-ZEL, α-ZEL, and ZEN were determined at 3.09, 25.90, 75.44, 97.44, and 98.15 in unstimulated cells, respectively, while in proliferating cells, the corresponding IC50 values were 0.73, 6.89, 45.16, 75.96, and 82.51. Neither viability nor proliferation was influenced by DOM-1 up to a concentration of 100 μM.The in vivo screening (N = 49) revealed the occurrence of ZEN (N = 24), α-ZEL (N = 3), β-ZEL (N = 37), DON, and DOM-1 (N = 2). The detected concentrations were much lower than the corresponding IC50 while the IC10 of DON and β-ZEL for proliferating PBMC corresponded to approximately 26 and 35 ng/mL which might be relevant when contaminated diets are fed.Clinical-chemical and hematological traits were not related to mycotoxin residue levels excepting blood urea nitrogen which was positively correlated to the sum of β-ZEL, α-ZEL, and ZEN concentration. Whether this reflects simply the feeding history of the horses or renal failures giving rise to a prolonged half-life of the toxins needs to be clarified further.

Topics: Animals; Blood Chemical Analysis; Cell Proliferation; Cell Survival; Female; Horses; Inhibitory Concentration 50; Leukocytes, Mononuclear; Male; Trichothecenes; Zearalenone

In this paper, a novel highly sensitive chemiluminescence immune-affinity 96 spots monolith array was developed to detect deoxynivalenol (DON), zearalenone (ZEN), T-2 toxin (T-2), and fumonisin B1 (FB1) in corn samples. Firstly, the monolith array was prepared through on suit UV-initiated copolymerization using polyethylene glycol diacrylate (PEGDA) as cross-linker, glycidyl methacrylate (GMA) as functional monomer and polyethylene glycol 200 (PEG 200) as the porogen. Subsequently, the four mycotoxins immune-affinity monolith array was prepared by immobilization of DON, ZEN, T-2, and FB1 antibody. The mole ratio of PEGDA/GMA, UV exposure time, and the volume ratio of PEG 200/PEGDA were optimized to improve the performances of the immune-affinity monolith array. For the mycotoxins immune-affinity monolith array based on chemiluminescence detection, the limit of detection was 0.0036ng/mL (DON), 0.0048ng/mL (ZEN), 0.0039ng/mL (T-2), and 0.0017ng/mL (FB1), respectively. The linear response in the range of 0.01-0.1ng/mL (R(2)=0.98). The results showed that the proposed four mycotoxins immune-affinity monolith array was a stable, accurate, and highly sensitive method to determine levels of DON, ZEN, T-2, and FB1 in real samples.

Topics: Antibodies, Immobilized; Equipment Design; Food Contamination; Food Microbiology; Fumonisins; Immunoassay; Limit of Detection; Luminescent Measurements; Protein Array Analysis; T-2 Toxin; Trichothecenes; Zea mays; Zearalenone

The aim of this study was to find effects of Fusarium toxins on brain injury in mice. We evaluated the individual and combined effect of the Fusarium toxins zearalenone and deoxynivalenol on the mouse brain. We examined brain weight, protein, antioxidant indicators, and apoptosis. After 3 and 5days of treatment, increased levels of nitric oxide, total nitric oxide synthase, hydroxyl radical scavenging, and malondialdehyde were observed in the treatment groups. This was accompanied by reduced levels of brain protein, superoxide dismutase (apart from the low-dose zearalenone groups), glutathione, glutathione peroxidase activity, and percentage of apoptotic cells. By day 12, most of these indicators had returned to control group levels. The effects of zearalenone and deoxynivalenol were dose-dependent, and were synergistic in combination. Our results suggest that brain function is affected by zearalenone and deoxynivalenol.

Topics: Animals; Antioxidants; Apoptosis; Brain; Dose-Response Relationship, Drug; Drug Synergism; Enzymes; Female; Fusarium; Malondialdehyde; Mice; Neurotransmitter Agents; Organ Size; Trichothecenes; Zearalenone

Fusarium genera can produce trichothecenes like deoxynivalenol (DON), zearalenone (ZEN) and T-2 toxin, which can occur in feed cereal grains. Enzyme-linked immunosorbent assays (ELISA) tests of different Hungarian swine feedstuff proved that these mycotoxins were present. In this survey, 45 feed samples from 3 significant Hungarian swine feedstuff manufacturers were tested. ELISA methodology validation showed mean recovery rates in ranges from 85.3% to 98.1%, with intermediate precision of 86.9-96.9% and variation coefficients of 3.4-5.7% and 5.9-7.1%, respectively. The results showed that among Fusarium toxins, generally DON was present in the highest concentration, followed by T-2 and finally ZEN in all tested swine feeds. Each of the mycotoxins was found above the limit of detection in all swine feedstuffs. Boars feed's DON (average ± standard deviation was 872 ± 139 µg kg

Topics: Animal Feed; Animal Husbandry; Animals; Edible Grain; Enzyme-Linked Immunosorbent Assay; Female; Food Contamination; Food Handling; Food Inspection; Hungary; Limit of Detection; Male; Poaceae; Reproducibility of Results; Statistics, Nonparametric; Sus scrofa; T-2 Toxin; Trichothecenes; Zearalenone

During an exposure, humans and animals are most often exposed to a mixture rather than individual mycotoxins. In this study, a Human Embryonic Kidney 293 cell (HEK-293) fluorescence sensor was developed to detect and evaluate mycotoxins, deoxynivalenol (DON) and zearalenone (ZEN) compounds, produced by Fusarium culmorum that are common food contaminants. TRE-copGFP (green fluorescent protein) and ERE-TagRFP (red fluorescent protein) plasmids were constructed and cotransfected into HEK-293 cells through a highly efficient, lipid-mediated, DNA-transfection procedure. Results show that fluorescence intensity was proportional to DON and ZEN concentrations, ranging from 2 to 40 ng/mL and 10 to 100 ng/mL respectively, with a detection limit of 0.75 ng/mL and 3.2 ng/mL respectively. The EC50 of DON and ZEN are 30.13 ng/mL and 76.63 ng/mL respectively. Additionally, ZEN may have a synergistic effect on enhancing AP-1 activity of the toxicity pathway of DON. These data indicate the high sensitivity and effectiveness of our biosensor system in the evaluation of the combined toxicity of ZEN, DON and their derivatives. In addition, this approach is suitable for an early warning method for the detection of ZEN and DON family mycotoxins contamination without higher-priced, conventional analytical chemistry methods.

Topics: Biosensing Techniques; Cell Separation; DNA; Flow Cytometry; Food Contamination; Fusarium; Green Fluorescent Proteins; HEK293 Cells; Humans; Lipids; Luminescent Proteins; Mycotoxins; Reactive Oxygen Species; Recombinant Proteins; Red Fluorescent Protein; Transfection; Trichothecenes; Zearalenone

The actual health risk from exposure to combined mycotoxins is unknown, and few studies have focused on changes to cellular biological systems (e.g., metabolomics) caused by combined mycotoxic effects. To evaluate the combined mycotoxic effects of deoxynivalenol (DON) and zearalenone (ZEN) on the level of cellular biological systems, gas chromatographic, time-of-flight mass spectroscopy (GC-TOF/MS) of the complete murine macrophage ANA-1 cell metabolome was implemented in this study. Using optimized chromatography and mass spectrometry parameters, the metabolites detected by GC-TOF/MS were identified and processed using multivariate statistical analysis, including principal component analysis (PCA) and orthogonal projection on latent-structures discriminant analysis (OPLS-DA). The metabolite sets were screened for further pathway analysis under rules of t-test (P) value < 0.05, VIP value > 1, and similarity value > 500. The mainly interfered metabolism pathways were categorized into two dominant types: amino acid metabolism and glycometabolism. Four metabolites, palmitic acid, 1-monopalmitin, ribose-5-phosphate and 2-deoxy-D-galactose, occur only under combined "DON + ZEN" treatment, indicating abnormal metabolism in ANA-1 cells. The metabolic state of ANA-1 cells under induction by combined "DON + ZEN" illustrates that DON may inhibit the estrogenic effects of ZEN. Thus, the combined effect of "DON + ZEN" may exacerbate toxicity in the pentose phosphate pathway, while palmitic acid metabolism is likely a new pathway effected by the combination, "DON + ZEN."

Topics: Animals; Cell Line; Gas Chromatography-Mass Spectrometry; Macrophages; Metabolomics; Mice; Trichothecenes; Zearalenone

Urinary biomarkers of mycotoxin exposure were evaluated in a group of celiac patients (

Topics: Adult; Biomarkers; Body Mass Index; Case-Control Studies; Celiac Disease; Diet; Diet, Gluten-Free; Female; Food Contamination; Food Microbiology; Fumonisins; Humans; Italy; Male; Middle Aged; Mycotoxins; Trichothecenes; Zearalenone

The purpose of this research was to investigate the toxicity of zearalenone (ZEA) on the growing performance, genital organs, serum hormones and histopathological changes of pre-pubertal female gilts, and to evaluate the efficacy of Bacillus subtilis ANSB01G in alleviating ZEA toxicosis in gilts. Eighteen pre-pubertal female gilts were randomly allocated to three treatments with one replicate per treatment. The gilts were fed following three diets for 24 days: the Control group was given a basic diet with normal corn; Treatment 1 (T1) was prepared by substituting corn naturally contaminated with ZEA for all normal corn in the basic diet (with a final concentrations of 238.57 μg kg(-1) of ZEA); and Treatment 2 (T2) was prepared by mixing the T1 diet with 2 kg T(-1) of fermented-dried culture of ANSB01G. The results showed that the presence of ZEA in diets significantly increased the vulva size and reproductive organ weight of the T1 gilts as compared with the Control group, and the addition of ANSB01G to diet naturally contaminated with ZEA obviously ameliorated these symptoms, as was observed in the T2 group. The presence of low doses of ZEA in the T1 diet had no significant effect on the level of follicle-stimulating hormone (FSH), luteotrophic hormone (LH) or serum oestradiol (E2) in the serum of gilts, but the prolactin (PRL) level in group T1 increased significantly. The gilts of the T1 group exhibited conspicuous cell enlargement and fatty degeneration of the corpus uteri, swelling, inflammation and lymphocyte infiltration of liver cells as compared with the Control group. The presence of ANSB01G can alleviate these hyperoestrogenic effects caused by ZEA, maintaining the body of gilt in a normal and healthy status. It is suggested that reproductive organs of gilts are seriously affected even if they are fed a low dose of ZEA in less time, and the addition of B. subtilis ANSB01G can effectively alleviate ZEA toxicosis in gilts.

Topics: Aflatoxins; Animal Feed; Animals; Bacillus subtilis; Estradiol; Estrogens, Non-Steroidal; Female; Follicle Stimulating Hormone; Food Contamination; Food Microbiology; Hormones; Liver; Mycotoxicosis; Organ Size; Prolactin; Swine; Trichothecenes; Vulva; Zea mays; Zearalenone

This study aimed to assess whether pathogenic Fusarium graminearum isolates from wheat and maize were more aggressive on their host of origin and whether aggressiveness was influenced further by B-trichothecene chemotype. Fifteen isolates were selected from a contemporary collection of isolates surveyed in New York in 2011 to 2012 to represent diversity of host of origin and chemotype. Three pathogenicity assays were used to evaluate and compare these isolates. Fusarium head blight (FHB) severity and trichothecene production in wheat, and maize seedling blight were evaluated in greenhouse inoculation experiments, and Gibberella ear rot (GER) severity and trichothecene production were evaluated in maize ears inoculated in the field. Our results showed among F. graminearum isolates a wide variation in aggressiveness and mycotoxin production toward wheat and maize and these isolates could not be structured by their host of origin or by chemotype. Moreover, aggressiveness rank order changed according to the host/organ evaluated. This indicates that relative susceptibility at the seedling stage may not predict susceptibility of ears. Significant correlations were observed of total trichothecenes (deoxynivalenol [DON] and its acetylated derivatives) produced with FHB and GER severity on wheat and maize, respectively. One isolate did not produce DON or ADON in wheat or maize kernels, yet was aggressive on both hosts. Nine of the fifteen isolates produced small amounts of zearalenone (ZON) in maize kernels, but not in wheat kernels, and ZON level was not correlated with GER severity. F. graminearum isolates from New York showed wide variation in aggressiveness and mycotoxin production toward susceptible wheat and maize. Neither host of origin nor trichothecene chemotype appeared to structure the populations we sampled.

Topics: DNA, Fungal; Fusarium; Genotype; Mycotoxins; New York; Plant Diseases; Seedlings; Seeds; Trichothecenes; Triticum; Virulence; Zea mays; Zearalenone

Deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEA) are mycotoxins commonly produced by Fusarium species. The purpose of the present study was to investigate the effects of DON alone and in combination with NIV and ZEA on several parameters including weight gain and histological aspects of pigs submitted to chronic intoxication. Twenty, 5-week-old piglets received for 28 days one of the following diets: a control diet, a diet mono- contaminated with DON (1.5mg/kg), a diet multi-contaminated with DON (2mg/kg)+NIV (1.3mg/kg)+ZEA (1.5mg/kg) or a diet contaminated with DON (3mg/kg)+NIV (1.3mg/kg)+ZEA (1.5mg/kg). Animals fed the multi-contaminated diets presented a significant decrease in weight gain over the total period. The chronic ingestion of the contaminated diets induced a significant increase on histological changes on the intestine, liver and lymphoid organs. In addition, a significant increase on lymphocyte apoptosis was observed in lymph nodes and spleen in the animals receiving the contaminated diets. These data provide a better understanding of the possible effects of Fusarium toxins, alone or in combinations on the morphology of the intestine and lymphoid organs, which would contribute to the risk assessment of these toxins.

Topics: Animal Feed; Animals; Cell Proliferation; Drug Interactions; Food Contamination; Intestines; Liver; Male; Swine; Trichothecenes; Weight Gain; Zearalenone

This study was performed to determine the individual and combined cytotoxic effects of Aflatoxin B1 (AFB1), zearalenone (ZEA), deoxynivalenol (DON) and fumonisin B1 (FB1) on BRL 3A rat liver cells. After the mycotoxins treated the BRL 3A cells for 12, 24 and 48 h, cell viability was determined using the MTT assay. The cytotoxicity of individual mycotoxins on BRL 3A cell viability in decreasing order were DON > AFB1 > ZEA > FB1. The central composite design (CCD) was used to assess the toxicity of binary and ternary mixtures of these mycotoxins. The mixtures of AFB1+ZEA and AFB1+DON showed the synergetic toxic effects on BRL 3A cells. These toxins decreased the viability of cells by inducing intracellular reactive oxygen species (ROS) production and promoting apoptosis in the BRL 3A cells. This effect was mediated by an upregulation of the stress and apoptotic genes Hsp70, p53, Bax, Caspase-3 and Caspase-8, along with a downregulation of the antiapoptotic gene Bcl-2. In conclusion, our results suggested that the coexistence of AFB1 and ZEA or DON in agricultural products could be more hepatotoxic than individually, suggests that the toxicological interactions of these toxins need to be better understood to assess health risks.

Topics: Aflatoxin B1; Animals; Apoptosis; bcl-2-Associated X Protein; Caspase 3; Caspase 8; Cell Line; Cell Survival; Down-Regulation; Fumonisins; Hepatocytes; Liver; Rats; Reactive Oxygen Species; Trichothecenes; Zearalenone

Sampling of straw bales from wheat, barley, and oats was carried out after harvest showing large variations in deoxynivalenol (DON) and zearalenone (ZEN) levels. In the wheat field, DON was detected in all straw samples with an average DON concentration of 976 μg/kg and a median of 525 μg/kg, while in four bales, the concentrations were above 3000 μg/kg. For ZEN, the concentrations were more uniform with an average concentration of 11 μg/kg. The barley straw bales were all positive for DON with an average concentration of 449 μg/kg and three bales above 800 μg/kg. In oat straw, the average DON concentration was 6719 μg/kg with the lowest concentration at 2614 μg/kg and eight samples above 8000 μg/kg. ZEN contamination was detected in all bales with an average concentration of 53 μg/kg with the highest concentration at 219 μg/kg. Oat bales from another field showed an average concentration of 16,382 μg/kg. ZEN concentrations in the oat bales were on average 153 μg/kg with a maximum at 284 μg/kg. Levels of Fusarium graminearum DNA were higher in oat straw (max 6444 pg DNA/mg straw) compared to straw from wheat or barley. The significance of mycotoxin exposure from straw should not be neglected particularly in years when high levels of DON and ZEN are also detected in the feed grain. With a limited number of samples preferably using a sampling probe, it is possible to distinguish lots of straw that should not be used as bedding material for pigs.

Topics: DNA, Fungal; Edible Grain; Food Contamination; Fusarium; Plant Stems; Trichothecenes; Zearalenone

Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins produced by fungi of the genus Fusarium which frequently contaminate maize and grain cereals. Mycotoxin-contaminated feed endangers animal health and leads to economic losses in animal production. Several mycotoxin elimination strategies, including the use of commercially available DON and ZEN detoxifying agents, have been developed. However, frequently there is no scientific proof of the efficacy of such adsorbents and degrading products. We therefore tested 20 commercially available products claiming to detoxify DON and/or ZEN either by biodegradation (4 products) or a combination of degradation and adsorption (16 products) under aerobic and anaerobic conditions at approx. pH 7. Under the applied conditions, a complete reduction of DON and consequent formation of the known non-toxic metabolite DOM-1 was exclusively observed in samples taken from the anaerobic degradation experiment of one product. For all other products, incubated under aerobic and anaerobic conditions, a maximum DON reduction of 17% after 72 h of incubation was detected. Aerobic and anaerobic incubation of only one tested product resulted in complete ZEN reduction as well as in the formation of the less-toxic metabolites DHZEN and HZEN. With this product, 68-97% of the toxin was metabolised within 3 h. After 24 h, a ZEN reduction ≥ 60% was obtained with four additional products during aerobic incubation only. Six of the 20 investigated products produced α- and/or β-ZEL, which are metabolites showing similar oestrogenic activity compared to ZEN. Aerobic and anaerobic degradation to unknown metabolites with unidentified toxicity was obtained with 10 and 3 products, respectively. The results of our study demonstrate the importance of in vitro experiments to critically screen agents claiming mycotoxin detoxification.

Topics: Edible Grain; Food Additives; Food Contamination; Fusarium; Tandem Mass Spectrometry; Trichothecenes; Zea mays; Zearalenone

A dose-response study was carried out to examine the carryover of zearalenone (ZEN), deoxynivalenol (DON) and their metabolites into bovine milk. Therefore, a feeding trial with 30 dairy cows fed with three different levels of Fusarium (FUS) toxin-contaminated maize was performed. A control group (0.02 mg ZEN kg(-1) dry matter (DM) and 0.07 DON kg(-1) DM) was compared with two groups fed contaminated diets. The first diet contained 0.33 mg ZEN kg(-1) DM and 2.62 mg DON kg(-1) DM (group FUS-50) and the second diet contained 0.66 mg ZEN kg(-1) DM and 5.24 mg DON kg(-1) DM (group FUS-100). For milk sample analysis, a new cost-efficient sample preparation method was developed for the simultaneous determination of ZEN, DON and their metabolites. The method comprised the separation of the milk fat followed by an SPE clean-up on Oasis HLB and a LC-MS/MS measurement. The less toxic metabolite de-epoxy-DON had the highest detected concentration (5.6 ng ml(-1) milk) in the milk samples obtained from the feeding trial. Additionally, ZEN (up to 0.29 ng ml(-1)), α-zearalenol (up to 0.17 ng ml(-1)), β-zearalenol (up to 0.95 ng ml(-1)) and DON (up to 2.5 ng ml(-1)) were detected in these samples. The milk toxin concentrations of cows fed the control diet were significantly lower compared with cows fed the contaminated diet. The calculated carryover rates ranged between 0 and 0.0075 for ZEN and metabolites and between 0 and 0.0017 for DON independent of exposure. It can be concluded that dietary toxin concentrations in the feed below or close to the current guidance values do not pose a risk for consumers due to negligible carryover rates.

Topics: Animal Feed; Animals; Cattle; Chromatography, Liquid; Female; Food Contamination; Fusarium; Milk; Solid Phase Extraction; Tandem Mass Spectrometry; Trichothecenes; Zea mays; Zearalenone

In this study, a novel and simple cell-based electrochemical biosensor was developed to assess the individual and combined toxicity of deoxynivalenol (DON) and zearalenone (ZEN) on BEL-7402 cells. The sensor was fabricated by modification with AuNPs, p-aminothiophenol, and folic acid in succession. The BEL-7402 cells which had a good activity were adhered on the electrode through the high affinity between the folate receptor and folic acid selectivity. We used the collagen to maintain the cell adhesion and viability. Electrochemical impedance spectroscopy (EIS) was developed to evaluate the individual and combined toxicity of DON and ZEN. Our results indicate that DON and ZEN caused a marked decrease in the cell viability in a dose-dependent manner. The value of electrochemical impedance spectroscopy decreased with the concentration of DON and ZEN in range of 0.1-20, 0.1-50 μg/ml with the detection limit as 0.03, 0.05 μg/ml, respectively, the IC50 for DON and ZEN as obtained by the proposed electrochemical method were 7.1 μg/ml and 24.6 μg/ml, respectively, and the combination of two mycotoxins appears to generate an additive response. The electrochemical cytotoxicity evaluation result was confirmed by biological assays. Compared to conventional methods, this electrochemical test is inexpensive, highly sensitive, and fast to respond, with long-term monitoring and real-time measurements. The proposed method provides a new avenue for evaluating the toxicity of mycotoxins.

Topics: Biological Assay; Cell Line, Tumor; Cell Survival; Dielectric Spectroscopy; Drug Combinations; Equipment Design; Equipment Failure Analysis; Humans; Lethal Dose 50; Mycotoxins; Neoplasms, Experimental; Reproducibility of Results; Sensitivity and Specificity; Toxicity Tests; Trichothecenes; Zearalenone

The objective of the presented study was to examine the influence of Fusarium mycotoxins (zearalenone--ZEN and deoxynivalenol--DON), administered separately and in combination, on the activity of cecal enzymes (β-glucosidase and β-glucuronidase) in gilts which were fed fodder con- taminated with these mycotoxins. The activity of β-glucosidase and β-glucuronidase varied in the range of 0.170-1.236 μmol · h(-1) · mg(-1) and 8.701-96.704 μmol · h(-1) · mg(-1), respectively. In the first two weeks, the toxins had no significant effect on the activity of β-glucosidase and β-glucuronidase in the ascending and descending colon. After week 3 and later on, ZEN and DON administered as a mix- ture led to the highest increase in the activity of both enzymes. Administered separately, DON affected the activity of enzymes more than ZEN. From the third week of the experiment, an increase in the activity of CW β-glucosidase and β-glucuronidase was observed.

Topics: Animal Feed; Animals; beta-Glucosidase; Cecum; Female; Fusarium; Gene Expression Regulation, Enzymologic; Mycotoxicosis; Swine; Swine Diseases; Trichothecenes; Zearalenone

This paper describes the use of two immunoaffinity columns (IACs) coupled in tandem, providing selective clean-up, based on targeted mycotoxins known to co-occur in specific matrices. An IAC for aflatoxins+ochratoxin A+fumonisins (AOF) was combined with an IAC for deoxynivalenol+zearalenone+T-2/HT-2 toxins (DZT); an IAC for ochratoxin A (O) was combined with a DZT column; and an aflatoxin+ochratoxin (AO) column was combined with a DZT column. By combining pairs of columns it was demonstrated that specific clean-up can be achieved as required for different matrices. Samples of rye flour, maize, breakfast cereal and wholemeal bread were analysed for mycotoxins regulated in the EU, by spiking at levels close to EU limits for adult and infant foods. After IAC clean-up extracts were analysed by LC-MS/MS with quantification using multiple reaction monitoring. Recoveries were found to be in range from 60 to 108%, RSDs below 10% depending on the matrix and mycotoxin combination and LOQs ranged from 0.1n g/g for aflatoxin B1 to 13.0 ng/g for deoxynivalenol. Surplus cereal proficiency test materials (FAPAS(®)) were also analysed with found levels of mycotoxins falling within the satisfactory range of concentrations (Z score ≤ ± 2), demonstrating the accuracy of the proposed multi-mycotoxin IAC methods.

Topics: Aflatoxins; Chemistry Techniques, Analytical; Chromatography, Liquid; Edible Grain; Food Analysis; Fumonisins; Mycotoxins; Ochratoxins; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

Mycotoxin producing moulds may contaminate numerous agricultural commodities either before harvest or during storage. A varied diet consisting of different foods may therefore be contaminated with a range of mycotoxins. The aim of the present study was to study concurrent exposure to mycotoxins through urinary multi-biomarker analysis, as well as its possible associations with the diet. Urinary samples from 252 adults, participating in the Swedish national dietary survey Riksmaten 2010-11, were collected together with a 4-day diet record. Concurrent mycotoxin exposure was studied using a multi-biomarker LC-MS/MS method. The results revealed that exposure to mycotoxins is common and concurrent exposure to more than one toxin was found in 69% of the study population. However, when comparing the number of toxins detected with the reported consumption data it was difficult to distinguish food patterns which would indicate an increased risk of exposure to many mycotoxins simultaneously. This is the first study to investigate concurrent mycotoxin exposure and urinary levels of fumonisin B1 (FB1), fumonisin B2 (FB2), nivalenol (NIV), ochratoxin A (OTA), zearalenone (ZEA), α-zearalenol (α-ZOL), β-zearalenol (β-ZOL) and de-epoxydeoxynivalenol (DOM-1) among adults in Sweden.

Topics: Adult; Aged; Asymptomatic Diseases; Biomarkers; Diet; Diet Records; Environmental Monitoring; Female; Food Contamination; Foodborne Diseases; Humans; Internet; Male; Middle Aged; Mycotoxins; Nutrition Surveys; Ochratoxins; Prevalence; Sweden; Trichothecenes; Zearalenone

This work evaluated the chemical composition and mycotoxin incidence in corn silage from 5 Brazilian dairy-producing regions: Castro, in central-eastern Paraná State (n=32); Toledo, in southwestern Paraná (n=20); southeastern Goiás (n=14); southern Minas Gerais (n=23); and western Santa Catarina (n=20). On each dairy farm, an infrared thermography camera was used to identify 3 sampling sites that exhibited the highest temperature, a moderate temperature, and the lowest temperature on the silo face, and 1 sample was collected from each site. The chemical composition and concentrations of mycotoxins were evaluated, including the levels of aflatoxins B1, B2, G1, and G2; zearalenone; ochratoxin A; deoxynivalenol; and fumonisins B1 and B2. The corn silage showed a highly variable chemical composition, containing, on average, 7.1±1.1%, 52.5±5.4%, and 65.2±3.6% crude protein, neutral detergent fiber, and total digestible nutrients, respectively. Mycotoxins were found in more than 91% of the samples, with zearalenone being the most prevalent (72.8%). All samples from the Castro region contained zearalenone at a high average concentration (334±374µg/kg), even in well-preserved silage. The incidence of aflatoxin B1 was low (0.92%). Silage temperature and the presence of mycotoxins were not correlated; similarly, differences were not observed in the concentration or incidence of mycotoxins across silage locations with different temperatures. Infrared thermography is an accurate tool for identifying heat sites, but temperature cannot be used to predict the chemical composition or the incidence of mycotoxins that have been analyzed, within the silage. The pre-harvest phase of the ensiling process is most likely the main source of mycotoxins in silage.

Topics: Aflatoxin B1; Brazil; Dairying; Food Contamination; Food Microbiology; Fumonisins; Infrared Rays; Mycotoxins; Ochratoxins; Silage; Surveys and Questionnaires; Thermography; Trichothecenes; Zea mays; Zearalenone

Maize is one of the most important crops and Poland is the fifth largest producing country in Europe. Diseases caused by Fusarium spp. can affect the yield and grain quality of maize because of contamination with numerous mycotoxins produced by these fungi. The present study was performed to identify the prevailing Fusarium species and the environmental factors affecting their frequencies and the contamination of grain with the main mycotoxins deoxynivalenol (DON), zearalenone (ZON) and fumonisin B1 (FB1). Thirty kernel samples were collected in three locations in 2011 and in seven locations in 2012 from three hybrids. On average, 25.24% kernels were colonized by Fusarium spp. (424 strains were isolated). Fusarium verticillioides and F. temperatum were the most prevalent species, F. subglutinans, F. proliferatum and F. graminearum were in minor abundance. In total, 272 isolates of F. verticillioides and 81 isolates of F. temperatum were identified. Fusarium temperatum frequency ranged from 1.70% to 28.57% and differences between locations were significant. Fumonisin B1 was found in all tested samples. DON was found in 66.67% and ZON in 43.33% of samples. Rainfall amount positively affected F. temperatum and F. subglutinans frequency in opposite to mean temperatures in July. On the other hand, relationships between frequency of these species and historical data from 1950-2000 for annual temperature range were negative in contrast to the coldest quarter temperatures.

Topics: Crops, Agricultural; Edible Grain; Environment; Europe; Food Contamination; Fumonisins; Fungi; Fusariosis; Fusarium; Mycotoxins; Poland; Temperature; Trichothecenes; Zea mays; Zearalenone

To elucidate the dietary exposure to deoxynivalenol (DON) and zearalenone (ZEN) from cereal-based products in Chinese populations using the probabilistic assessment approach.. A total of 292 wheat flours and 347 corn-based products were collected from sampling sites of 107 supermarkets or farmers markets, which were randomly selected from 44 cities of 13 provinces in 2009 by the stratified cluster random sampling method. Then, DON and ZEN contamination levels in these samples above analyzed by UPLC-MS/MS in combination with the food consumption data of 68 959 respondents, who were divided into group 1 aged 3 to 13 years old, and group 2 aged 14 and over 14 years old (≥14 years old), obtained by China National Nutrition and Health Survey in 2002 were investigated. A probabilistic assessment model using Monte Carlo simulation was applied to derive the intake distribution of P(1)-P(99) percentile of dietary exposure to DON and ZEN. Meanwhile, all parameters related to dietary exposure to both toxins were compared with either the provisional maximum tolerable daily intake (PMTDI) of 1 µg·kg(-1)·d(-1) for DON, or the tolerable daily intake (TDI) of 0.25 µg·kg(-1)·d(-1) for ZEN in order to evaluate the risk of dietary intake of two toxins and find the minimum percentile of dietary exposure to these two toxins. The statistical differences of dietary exposure to these two toxins between two groups were achieved by t test.. The detection frequencies of DON in wheat flours and corn-based products were 100% (292/292) and 97.4% (338/347), respectively. A total of 21 out of 639 samples (wheat flours: 5/292, corn-based products: 16/347) were positive for DON at the levels exceeding the Chinese regulatory limit of 1 000 µg/kg for DON. And the detection frequencies of ZEN in wheat flours and corn-based products were 53.4% (156/292) and 87.6% (304/347), respectively.54 out of 347 corn-based products and no wheat flours were positive for ZEN at the levels exceeding the Chinese regulatory limit of 60 µg/kg for ZEN. Meanwhile, the mean values (95% CI) of the P(50), P(75), P(90), P(95), P(97.5) and P(99) percentile of dietary exposure to DON in populations of 3 to 13 years old were 0.170 (0.170-0.171), 0.762 (0.759-0.765), 2.066 (2.038-2.069), 3.515 (3.501-3.530), 5.342 (5.314-5.372), and 9.220 (9.155-9.279) µg · kg(-1)·d(-1), which were higher than those in populations of ≥14 years old (0.131 (0.130-0.131), 0.500 (0.498-0.501), 1.280 (1.276-1.285), 2.138 (2.128-2.14), 3.510 (3.494-3.527), and 5.512 (5.474-5.546) µg·kg(-1)·d(-1)), with t values of 87.19, 163.87, 164.66, 157.78, 105.47 and 96.31, and all P values less than 0.001. And the mean values (95% CI) of the P(50), P(75), P(90), P(95), P(97.5) and P(99) percentile of dietary exposure to ZEN in populations of 3 to 13 years old were 0.001 (0.001-0.001), 0.006 (0.006-0.006), 0.039 (0.038-0.039), 0.101 (0.100-0.101), 0.195 (0.194-0.197) and 0.378 (0.374-0.381) µg · kg(-1)·d(-1), which were also higher than those in populations of ≥14 years old (0.001 (0.001-0.001), 0.004 (0.004-0.004), 0.026 (0.026-0.026), 0.061 (0.060-0.061), 0.115 (0.115-0.116) and 0.232 (0.231-0.235) µg·kg(-1)·d(-1)) with T-values of 151.11, 73.80, 96.81, 100.81, 91.93 and 76.13, and all P values less than 0.001. Besides, the minimum percentile of dietary exposure to DON in populations of 3 to 13 years old and ≥14 years old exceeded the corresponding PMTDI of 1 µg·kg(-1)·d(-1) was found in the probability distribution of P(76) (99% percentile = 1.03 µg·kg(-1)·d(-1)) and P(84) (95% percentile = 1.01 µg·kg(-1)·d(-1)) percentile, respectively. And the minimum percentile of dietary exposure to ZEN in populations of 3 to 13 years old and ≥14 years old exceeded the corresponding TDI of 0.25 µg·kg(-1)·d(-1) was found in the probability distribution of P(97) (95% percentile = 0.25 µg·kg(-1)·d(-1)) and P(98) (90% percentile = 0.26 µg·kg(-1)·d(-1)) percentile, respectively.. The contamination levels of DON and ZEN in wheat flours and corn-based products and the risk of dietary exposure to both DON and ZEN in populations in Chinese populations were at relatively low levels. The dietary exposure to both DON and ZEN in populations of 3 to 13 years old was higher than those in populations of ≥14 years old . Populations of 3 to 13 years old were the populations at the high risk of dietary exposure to both mycotoxins.

Topics: Adolescent; Adult; Child; Child, Preschool; China; Diet; Edible Grain; Food Contamination; Humans; Middle Aged; Mycotoxins; Tandem Mass Spectrometry; Trichothecenes; Triticum; Zea mays; Zearalenone

Field experiments were conducted to identify the impact of post-anthesis rainfall on the concentration of deoxynivalenol (DON) and zearalenone (ZON) in harvested wheat grain. Winter wheat plots were inoculated with Fusarium graminearum at stem extension (GS31) and prothioconazole was applied at mid-anthesis (GS65) to split plots and plots were subsequently mist irrigated for 5 days. Plots were either covered by polytunnels, irrigated by sprinklers or left as non-irrigated uncovered control plots after medium-milk (GS75). Plots were harvested either when ripe (GS92; early harvest) or three weeks later (late harvest). Fusarium head blight (FHB) was assessed each week from inoculation. At harvest, yield and grain quality was measured and grains were analysed for DON and ZON. Differences in rainfall resulted in contrasting disease pressure in the two experiments, with low FHB in the first experiment and high FHB in the second. Difference in FHB resulted in large differences in grain yield, quality and mycotoxin content. DON concentration was significantly (P < 0.05) higher in irrigated compared to covered and control plots in the first experiment, whereas in the second experiment, DON was significantly (P < 0.05) higher in the covered plots compared to the control and irrigated plots. ZON concentration was significantly (P < 0.05) higher in irrigated plots in both experiments. Later harvesting resulted in an approximate fivefold increase in ZON in the first experiment, but was not significantly different in the second experiment. Prothioconazole significantly (P < 0.05) reduced DON in both experiments, but gave inconsistent reductions to ZON. This is the first report to show that the post-anthesis rainfall can significantly increase ZON in wheat, which can increase further with a delayed harvest but may be significantly reduced with the application of prothioconazole. Importantly, in the absence of moisture late season, ZON remains at very low concentrations even when wheat is severely affected by FHB.

Topics: Animal Feed; Animals; Edible Grain; Food Contamination; Fungicides, Industrial; Fusarium; Mycotoxins; Plant Diseases; Rain; Time Factors; Triazoles; Trichothecenes; Triticum; Zearalenone

This study was performed to investigate the individual and combined toxic effects of deoxynivalenol (DON) and zearalenone (ZEA) on mouse kidney. A total of 360 female mice were divided into nine groups. Each group received intraperitoneal injection of solvent (control), DON, ZEA, or DON+ZEA four times for 12d. Results showed that ZEA and/or DON increased the apoptosis rate in the kidney, as well as the levels of serum creatinine and blood urea nitrogen. DON and/or ZEA also induced renal oxidative stress as indicated by increased malondialdehyde concentration and nitric oxide level and reduced superoxide dismutase enzyme activity and hydroxyl radical inhibiting capacity. The observed changes were dose and time dependent. This study reports that DON and/or ZEA induced apoptosis, dysfunction, and oxidative stress in mouse kidney. Furthermore, the combination of DON+ZEA exhibited a sub-additive nephrotoxic effect.

Topics: Animals; Apoptosis; Blood Urea Nitrogen; Creatinine; Dose-Response Relationship, Drug; Female; Injections, Intraperitoneal; Kidney; Mice; Oxidative Stress; Trichothecenes; Zearalenone

Aflatoxins, ochratoxins, fumonisins, deoxynivalenol, and zearalenone are of significant public health concern as they can cause serious adverse effects in different organs including the liver, kidney, and immune system in humans. These toxic secondary metabolites are produced by filamentous fungi mainly in the genus Aspergillus, Penicillium, and Fusarium. It is challenging to control the formation of mycotoxins due to the worldwide occurrence of these fungi in food and the environment. In addition to raw agricultural commodities, mycotoxins tend to remain in finished food products as they may not be destroyed by conventional processing techniques. Hence, much of our concern is directed to chronic health effects through long-term exposure to one or multiple mycotoxins from contaminated foods. Ideally risk assessment requires a comprehensive data, including toxicological and epidemiological studies as well as surveillance and exposure assessment. Setting of regulatory limits for mycotoxins is considered necessary to protect human health from mycotoxin exposure. Although advances in analytical techniques provide basic yet critical tool in regulation as well as all aspects of scientific research, it has been acknowledged that different forms of mycotoxins such as analogs and conjugated mycotoxins may constitute a significant source of dietary exposure. Further studies should be warranted to correlate mycotoxin exposure and human health possibly via identification and validation of suitable biomarkers.

Topics: Aflatoxins; Aspergillus; Environmental Exposure; Food Contamination; Fumonisins; Fungi; Fusarium; Humans; Mycotoxins; Ochratoxins; Penicillium; Public Health; Trichothecenes; Zearalenone

Immature gilts were administered per os with zearalenone (ZEN) at 40 μg/kg BW (group Z, n = 9), deoxynivalenol (DON) at 12 μg/kg BW (group D, n = 9), a mixture of ZEN and DON (group M, n = 9) or a placebo (group C, n = 9) over a period of six weeks. The pigs were sacrificed after one, three, or six weeks of the treatment (12 pigs per each time-point). Histological investigations revealed an increase in the mucosal thickness and the crypt depth as well as a decrease in the ratio of the villus height to the crypt depth in groups D and M after six weeks of exposure to the mycotoxins. The number of goblet cells in the villus epithelium was elevated in groups Z and M after one week and in group D after three weeks. The administration of ZEN increased the lymphocyte number in the villus epithelium after 1 week and the plasma cell quantity in the lamina propria after one, three, and six weeks of the experiment. DON treatment resulted in an increase in the lymphocyte number in the villus epithelium and the lamina propria after six weeks, and in the plasma cell quantity in the lamina propria after one, three, and six weeks of exposure. In group M, lymphocyte counts in the epithelium and the lamina propria increased significantly after six weeks. Neither mycotoxin induced significant adverse changes in the ultrastructure of the mucosal epithelium and the lamina propria or in the intestinal barrier permeability. Our results indicate that immune cells are the principal target of low doses of ZEN and DON.

Topics: Animals; Female; Fusarium; Goblet Cells; Humans; Intestinal Mucosa; Jejunum; Lymphocyte Count; Lymphocytes; Mycotoxins; Plasma Cells; Sus scrofa; Trichothecenes; Zearalenone

Organic farming does not allow the use of conventional mineral fertilizers and crop protection products. As a result, in our experiments we chose to grow different species of cereals and to see how cereal species affect mycotoxin accumulation. This study describes the occurrence of deoxynivalenol (DON), zearalenone (ZEA) and T-2/HT-2 toxin in a survey of spelt and common wheat and their bran as well as flour. The analysis was conducted using an enzyme-linked immunosorbent assay (ELISA) method. The concentrations of DON, ZEA and T-2/HT-2 in Triticum spelta and T. aestivum were influenced by species, cereal type and year interaction. The highest concentrations of these mycotoxins were found in spelt grain with glumes, in spelt glumes and in spring wheat. These results show significantly higher concentrations of Fusarium toxins in glumes than in dehulled grain, which indicates the possible protective effect of spelt wheat glumes. The lowest DON, ZEA and T-2/HT-2 concentrations were determined in spelt grain without glumes. The research shows that it is potentially risky to produce bran from grain in which mycotoxin concentrations are below limits by European Union Regulation No. 1881/2006, since the concentration of mycotoxins in bran can be several times higher than that in grain. As a result, although bran is a dietary product characterised by good digestive properties, it can become a harmful product that can cause unpredictable health damage.

Topics: Dietary Fiber; Enzyme-Linked Immunosorbent Assay; European Union; Flour; Food Contamination; Food Microbiology; Food Safety; Food, Organic; Fusarium; Hazard Analysis and Critical Control Points; Humans; Mycotoxins; Organic Agriculture; T-2 Toxin; Trichothecenes; Triticum; Weather; Zearalenone

A feeding trial with dairy cows fed graded proportions of a Fusarium toxin contaminated maize containing mainly deoxynivalenol (DON) was carried out to relate the plasma levels of DON, zearalenone (ZEN) and their metabolites to the performance. German Holstein cows (n=30) were divided into three groups (n=10 in each): CON (0.02mgZEN and 0.07mgDON, per kg dry matter, DM), FUS-50 (0.33mg ZEN and 2.62mgDON, per kg DM), FUS-100 (0.66mgZEN and 5.24mgDON, per kg DM). The average performance level was characterised by daily DM intake, energy balance and milk yield which were not affected by the DON and ZEN levels in feed. DON, de-epoxy-DON (de-DON) and ZEN were detected simultaneously in all plasma samples. A linear relationship between toxin intake and plasma levels could be established. Moreover, a linear relationship between DON and de-DON concentration could be derived. It was concluded that DON and ZEN intake of 0.5mgZEN/kg and 5mgDON/kg (current guidance values) had no considerable effects on the performance parameter of dairy cows. Furthermore, increased plasma concentrations of ZEN, DON and de-DON may hint on toxin exposure through the diets.

Topics: Animal Feed; Animals; Cattle; Dairying; Fusarium; Trichothecenes; Zea mays; Zearalenone

Clonostachys rosea strain IK726 is a mycoparasitic fungus capable of controlling mycotoxin-producing Fusarium species, including F. graminearum and F. culmorum, known to produce Zearalenone (ZEA) and Deoxynivalenol (DON). DON is a type B trichothecene known to interfere with protein synthesis in eukaryotes. ZEA is a estrogenic-mimicing mycotoxin that exhibits antifungal growth. C. rosea produces the enzyme zearalenone hydrolase (ZHD101), which degrades ZEA. However, the molecular basis of resistance to DON in C. rosea is not understood. We have exploited a genome-wide transcriptomic approach to identify genes induced by DON and ZEA in order to investigate the molecular basis of mycotoxin resistance C. rosea.. We generated DON- and ZEA-induced cDNA libraries based on suppression subtractive hybridization. A total of 443 and 446 sequenced clones (corresponding to 58 and 65 genes) from the DON- and ZEA-induced library, respectively, were analysed. DON-induced transcripts represented genes encoding metabolic enzymes such as cytochrome P450, cytochrome c oxidase and stress response proteins. In contrast, transcripts encoding the ZEA-detoxifying enzyme ZHD101 and those encoding a number of ATP-Binding Cassette (ABC) transporter transcripts were highly frequent in the ZEA-induced library. Subsequent bioinformatics analysis predicted that all transcripts with similarity to ABC transporters could be ascribed to only 2 ABC transporters genes, and phylogenetic analysis of the predicted ABC transporters suggested that they belong to group G (pleiotropic drug transporters) of the fungal ABC transporter gene family. This is the first report suggesting involvement of ABC transporters in ZEA tolerance. Expression patterns of a selected set of DON- and ZEA-induced genes were validated by the use of quantitative RT-PCR after exposure to the toxins. The qRT-PCR results obtained confirm the expression patterns suggested from the EST redundancy data.. The present study identifies a number of transcripts encoding proteins that are potentially involved in conferring resistance to DON and ZEA in the mycoparasitic fungus C. rosea. Whilst metabolic readjustment is potentially the key to withstanding DON, the fungus produces ZHD101 to detoxify ZEA and ABC transporters to transport ZEA or its degradation products out from the fungal cell.

Topics: ATP-Binding Cassette Transporters; Drug Resistance, Fungal; Expressed Sequence Tags; Fungal Proteins; Fusarium; Gene Expression Profiling; Gene Expression Regulation, Fungal; Gene Library; Genome, Fungal; Hypocreales; Mycotoxins; Trichothecenes; Zearalenone

This study comprises analyses of contents of mycotoxins, such as deoxynivalenol and zearalenone, as well as the level of oxidative stress in ears of a susceptible wheat cultivar Hanseat and cv. Arina, resistant to a pathogenic fungus Fusarium graminearum. Starting from 48 h after inoculation, a marked increase was observed in the contents of these mycotoxins in ears of wheat; however, the greatest accumulation was recorded in the late period after inoculation, i.e., during development of disease. Up to 120 h after inoculation, in ears of both wheat cultivars, the level of deoxynivalenol was higher than that of zearalenone. The susceptible cultivar was characterized by a much greater accumulation of deoxynivalenol than the resistant cultivar. At the same time, in this cultivar, in the time from 0 to 72 h after inoculation, a marked post-infection increase was observed in the generation of the superoxide radical (O2•-). Additionally, its level, at all the time points after inoculation, was higher than in the control. In wheat cv. Arina, a markedly higher level of O2•- generation in relation to the control was found up to two hours after inoculation and, next, at a later time after inoculation. In turn, the level of semiquinone radicals detected by electron paramagnetic resonance (EPR) increased at later culture times, both in cv. Hanseat and Arina; however, in infested ears of wheat, it was generally lower than in the control. Analysis of disease symptoms revealed the presence of more extensive lesions in ears of a susceptible wheat cv. Hanseat than resistant cv. Arina. Additionally, ergosterol level as a fungal growth indicator was higher in ears of susceptible wheat than in the resistant cultivar.

Topics: Ergosterol; Fusarium; Oxidative Stress; Quinones; Superoxides; Trichothecenes; Triticum; Zearalenone

Fumonisin B1 (FB1) is a Fusarium mycotoxin frequently occurring in corn in combination with deoxynivalenol (DON) and zearalenone. The aim of this study was to determine if FB1, alone and combined with DON or α-zearalenol (ZEA), zearalenone major active metabolite, can affect granulosa cell proliferation, steroid production, and gene expression in swine. Porcine granulosa cells were cultured for 2 days in serum-containing medium followed by 1 or 2 days in serum-free medium with or without added treatments. Fumonisin B1 had inhibitory effects on granulosa cell proliferation. Deoxynivalenol strongly inhibited cell growth, and no significant difference was detected in combination with FB1. α-Zearalenol showed a stimulatory effect on granulosa cell numbers even in combination with FB1. Regarding steroid production, FB1 increased progesterone production, and FB1 had no effect on estradiol production. Deoxynivalenol strongly inhibited progesterone and estradiol production, and FB1 had no significant effect on this response. α-Zearalenol increased progesterone production, and its combination with FB1 produced additive effects. α-Zearalenol had no effect on estradiol production, whereas it decreased estradiol production when co-treated with FB1. Fumonisin B1 was found to decrease CYP11A1 messenger RNA abundance, and the stimulatory effect of FB1 on progesterone production was found to be not dependent on 3-hydroxy-3-methylglutaryl-coenzyme A reductase activity suggesting that FB1 increases progesterone production through a different mechanism. The results show that these Fusarium mycotoxins can influence porcine granulosa cell proliferation and steroid production, thereby demonstrating their potential reproductive effects on swine.

Topics: Animals; Cell Proliferation; Cells, Cultured; Cholesterol Side-Chain Cleavage Enzyme; Dose-Response Relationship, Drug; Drug Interactions; Estradiol; Female; Follicle Stimulating Hormone; Fumonisins; Gonadal Steroid Hormones; Granulosa Cells; Hydroxymethylglutaryl-CoA Reductase Inhibitors; Insulin-Like Growth Factor I; Progesterone; RNA, Messenger; Simvastatin; Swine; Trichothecenes; Zearalenone

Improperly practiced postharvest procedures can pose mycotoxin-related risks in the production of medicinal herbs. As a health food with pharmacological supplements, cereal-based adlay has been broadly used in oriental medical practice. Compared with the standard production protocol, three provisional critical control points (CCPs) in the conventional procedure were identified and assessed for mycotoxin contamination in the adlay from small farms in Korea. Although various mycotoxins were present, the prevalence of deoxynivalenol (DON) or zearalenone (ZEN) was relatively high in the adlay. In terms of drying conditions, field drying in the conventional pathway was associated with more exposure to DON than heated-air drying. Moreover, the DON or ZEN levels in chaff were higher than the levels in the inner grain, suggesting that the hulling process as another CCP would reduce the DON or ZEN exposure. In particular, the DON or ZEN levels in adlay stored for protracted periods without dehulling were very high, but a lower storage temperature of 12°C was not effective at significantly reducing these mycotoxins. In this case, the inner grain was more contaminated with DON or ZEN than the chaff after protracted storage because surface fungi, which produce mycotoxins, can penetrate deep into grain with time. Heated-air drying and nonprotracted storage limited DON contamination in adlay. More importantly, an early dehulling process should be adopted as an easy preventive action to reduce the risk of exposure to DON or ZEN in adlay postharvest. This is monitored as a central CCP for safer production of adlay from local farms.

Topics: Coix; Dietary Supplements; Food Contamination; Food Handling; Republic of Korea; Trichothecenes; Zearalenone

A three-year (2010-2012) survey was conducted to assess the prevalence and concentrations of deoxynivalenol (DON) and zearalenone (ZEN) in wheat from several regions of Jiangsu province, China, which are heavily impacted by Fusarium head blight. A total of 180 wheat samples were obtained from the infected fields that spread 21 counties. DON and ZEN levels were determined by liquid chromatography-mass spectrometry (LC-MS/MS). DON was found in 74.4% of samples at levels ranging from 14.52 to 41157.13 μg/kg (mean 488.02 μg/kg), while ZEN was found in 12.8% of samples at levels ranging from 10.13 to 3048.88 μg/kg (mean 73.04 μg/kg). In years and regions of higher rainfall, DON and ZEN levels were higher in samples. These results are necessary to take a vigilant attitude to prevent human intake of trichothecenes and protect human's health from the risk of exposure to these toxins.

Topics: China; Fusarium; Humans; Trichothecenes; Triticum; Zearalenone

It is expected that humans are exposed to combined mycotoxins, which occur simultaneously in the food items, than to individual compounds and that can increase their potential toxicity. Considering this coincident production, deoxynivalenol (DON) and zearalenone (ZEN) as they are produced by several Fusarium species, can interfere at a cellular level. Therefore, these two toxins were chosen to study their interactive effects on human colon carcinoma cells (HCT116), using the endpoints including cell viability, cell cycle analysis, mitochondrial transmembrane potential (ΔΨm) determination and permeability transition pore (PTP) opening. Our results showed that DON and ZEN caused a marked decrease of cell viability in a dose-dependent manner, mediated by an activation of the mitochondrial apoptotic process; characterized by PTP opening and the loss of ΔΨm. Nevertheless, combined DON and ZEN reduced all the toxicities observed with the mycotoxins separately. Therefore, the combination of the two mycotoxins appears as a sub-additive response.

Topics: Apoptosis; Cell Death; Cell Survival; HCT116 Cells; Humans; Membrane Potential, Mitochondrial; Mitochondrial Membrane Transport Proteins; Mitochondrial Permeability Transition Pore; Mycotoxins; Trichothecenes; Zearalenone

A new lateral flow immunoassay (LFA) is proposed for qualitative and/or semiquantitative determination of aflatoxin B1 (AFB1), zearalenone (ZEA), deoxynivalenol (DON), and their analogues (AFs, ZEAs, DONs) in cereal samples. Each of the mycotoxin specific antibody was class specific and there was no cross reactivity to other groups of compounds. The visual limits of detection (vLOD) of the strip were 0.03, 1.6, and 10 μg/kg for AFB1, ZEA and DON, respectively. The calculated limits of detection (cLOD) were 0.05, 1, and 3 μg/kg, respectively. Meanwhile the cutoff values were achieved at 1, 50, and 60 μg/kg for AFB1, ZEA and DON, respectively. Recoveries ranged from 80% to 122% and RSD from 5% to 20%. Both the vLOD and cLOD for the three mycotoxins were lower than the EU maximum levels. Analysis of naturally contaminated maize samples resulted in a good agreement between the multiplex LFA and LC-MS/MS (100% for DONs and AFs, and 81% for ZEAs). Careful analysis of the results further explained the general overestimation of LFA compared to chromatographic methods for quantification of mycotoxins.

Topics: Aflatoxin B1; Antibody Specificity; Immunoassay; Limit of Detection; Mycotoxins; Tandem Mass Spectrometry; Trichothecenes; Triticum; Zea mays; Zearalenone

A rapid method has been developed for the determination of aflatoxins B1, B2, G1 and G2, ochratoxin A, zearalenone, T-2 toxin, deoxynivalenol and fumonisin B1 in cereal-based complementary foods for infants and young children. The mycotoxins were extracted from the samples using a modified QuEChERS-based extraction procedure without any further clean-up step. The separation of the analytes was carried out on an Agilent XDB C18 column (50 mm x 2.1 mm, 1.8 microm) using the mobile phases of acetonitrile and 5 mmol/L ammonium acetate-0.1% (volume percentage) formic acid aqueous solution with gradient elution. The extract was determined by rapid resolution liquid chromatography-tandem mass spectrometry. Matrix-matched calibration was used for the quantification. The proposed method showed good linear correlation with the correlation coefficients all above 0.98. The blank samples were fortified at three levels, and the recoveries ranged from 77.6% to 105.7% with the RSDs from 2.5% to 13.7%. The limits of detection ranged from 0.1 microg/kg to 15.8 microg/kg. The developed method was successfully applied for mycotoxin analysis in 41 samples bought from markets. The method shows advantages in both accuracy and sensitivity, and is suitable for the rapid multi-aflatoxin screening in cereal-based complementary foods for infants and young children.

Topics: Acetonitriles; Aflatoxin B1; Chromatography, Liquid; Edible Grain; Fumonisins; Humans; Infant; Infant Food; Mass Spectrometry; Mycotoxins; Ochratoxins; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

The occurrence of the most widespread type A and B trichothecenes and of zearalenone was surveyed in soft and durum wheat produced in northern Italy. A total of 293 wheat fields, grown in the years 2009-2011, were surveyed; for each field, weather and cropping system data were collected. The results indicated a high deoxynivalenol incidence, with durum always more contaminated than soft wheat; in 2010, the percentage of durum wheat samples exceeding the European Commission legal limit was 39.6%. As regards type A trichothecenes, widespread contamination was observed in 2010. In soft wheat, an incidence of 70% and 85% was found for T-2 and HT-2 toxins, respectively; all the durum wheat samples were contaminated. The trichothecene contamination was affected by weather conditions; copious rainfall and high relative humidity (RH) during flowering occurred in 2010, when the highest contamination of both type A and B trichothecenes was found.

Topics: Food Contamination; Fusarium; Humans; Italy; Seeds; T-2 Toxin; Trichothecenes; Triticum; Weather; Zearalenone

A sensitive and selective liquid chromatography tandem mass spectrometry method using negative electrospray ionisation (LC-ESI-MS/MS) was developed for the simultaneous determination of zearalenone (ZEN), deoxynivalenol (DON) and their metabolites α-zearalenol, β-zearalenol, zearalanone, α-zearalanol, β-zearalanol and de-epoxy-deoxynivalenol in pig serum. For method development, different sample preparation columns were tested for their suitability for extraction and clean up. Finally, preparation of serum samples was carried out using Oasis™ HLB solid-phase extraction (SPE) columns. The analyte concentrations were determined by the use of isotopically labelled internal standards (IS). The method was in-house validated for all analytes. Calibration graphs (0.3-480 ng/ml) were prepared and high degree of linearity was achieved (r ≥ 0.99). Results for method precision ranged between 2.7 and 21.5 % for inter-day and between 1.1 and 11.1 % for intra-day. The recoveries were in the range of 82-131 %. Limits of detection and quantification ranged 0.03-0.71 and 0.08-2.37 ng/ml, respectively. The method has been successfully used for quantitative determination of ZEN, DON and their metabolites in pig serum from a feeding trial with practically relevant ZEN and DON concentrations. This method is precise and reproducible and can be used as a multi-biomarker method to assess animal exposure to these mycotoxins and for diagnosis of intoxications.

Topics: Animals; Chromatography, Liquid; Female; Limit of Detection; Metabolomics; Reproducibility of Results; Swine; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

The differential risk of exposure to fumonisin (FB), deoxynivalenol (DON), and zearalenone (ZEA) mycotoxins to the South African population, residing in the nine Provinces was assessed during a cross-sectional grain consumer survey. The relative per capita maize intake (g/day) was stratified by gender, ethnicity, and Province and the probable daily intake (PDI) for each mycotoxin (ng/kg body weight/day) calculated utilizing SPECIAL and SUPER dry milled maize fractions representing different exposure scenarios. Men consumed on an average more maize (173 g/day) than women (142 g/day) whereas the black African ethnic group had the highest intake (279 g/day) followed by the Colored group (169 g/day) with the Asian/Indian and White groups consuming lower quantities of 101 and 80 g/day, respectively. The estimated mean PDIs for the various subgroups and Provinces, utilizing the different dry milled maize fractions, were below the provisional maximum tolerable daily intake (PMTDI) for each mycotoxin. A distinct and more sensitive mycotoxin risk assessment model (MYCORAM) for exposure, stratified by Province and ethnicity were developed utilizing specific maize intake increments (g/kg body weight/day) that provides information on the percentage of the population exposed above the PMTDI for each mycotoxin. Evaluation of the MYCORAM utilizing commercial and EXPERIMENTALLY DERIVED: SPECIAL milling fractions, containing predefined mycotoxins levels, predicts the percentage of maize consumers exposed above the respective PMTDI. Safety modeling using the MYCORAM could also predict a maximum tolerated level adequate to safeguard all South African maize consumers including the most vulnerable groups.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Body Weight; Cross-Sectional Studies; Eating; Ethnicity; Feeding Behavior; Female; Food Handling; Food Microbiology; Fumonisins; Humans; Male; Middle Aged; Residence Characteristics; Risk Assessment; Risk Factors; Sex Factors; South America; Surveys and Questionnaires; Trichothecenes; Young Adult; Zea mays; Zearalenone

The contamination of plant material with mycotoxins, in particular of the genus Fusarium, is common in the natural environment. Multiparous female wild boars are exposed to feed contaminated with zearalenone (ZEN) and deoxynivalenol throughout the year. The aim of this study was to determine the concentrations of the above mycotoxins in multiparous female wild boars and to describe their effect on the histological structure of the ovaries at the beginning of astronomical winter. Toxicological examinations revealed 0.291 ng/ml of ZEN, 0.406 ng/ml of α-zearalenol (α-ZEL), 0.392 ng/ml β-zearalenol (β-ZEL) and an absence of deoxynivalenol (values below the sensitivity of the method) in the blood plasma of multiparous female wild boars. Numerous ovarian follicles at various stages of development, characterized by different degree of damage, were observed. Numerous deformed resting ovarian follicles were noted directly under the epithelium, and signs of follicular atresia and hyalinization were observed. Blood vessels in the medulla of the ovary were dilated, which probably improved the distribution of ZEN in the ovaries. Higher substrate (ZEN) concentrations in the ovaries led to an insignificant increase in the staining intensity of 3β-HSD and 17β-HSD clusters. The observed changes could contribute to prolonging the initial stage of late anestrus in multiparous female wild boars.

Topics: Animals; Endocrine Disruptors; Environmental Monitoring; Estrous Cycle; Female; Mycotoxins; Ovary; Seasons; Sus scrofa; Trichothecenes; Zearalenone

The objective of the study was to determine the effect of exposure of pigs to the Fusarium mycotoxins zearalenone (ZEN) and deoxynivalenol (DON), administered together and separately, on the colon microbiota. An experiment was conducted for 42 days on gilts, randomly assigned to four groups and administered either ZEN, DON, ZEN+DON, or a placebo. The number of aerobic mesophilic bacteria, yeasts, molds, anaerobic Clostridium perfringens, fecal streptococci, Enterobacteriaceae, Escherichia coli, and lactic acid bacteria (LAB) were determined in the contents of the ascending colon. The influence of mycotoxins on the functional diversity of the colonic microbiota was assessed using EcoPlate tests (Biolog). Analysis revealed the predominance of LAB in all groups of pigs. Zearalenone, administered separately and together with DON, was found to have an adverse effect on mesophilic aerobic bacteria, but only after long exposure to this mycotoxin. During the six weeks of the experiment, the concentration of C. perfringens, E. coli, and other bacteria in the family Enterobacteriaceae was most considerably reduced in the experimental groups exposed to zearalenone, both separately and together with DON. Mycotoxins also affected the functional biodiversity of microorganisms. Both Shannon's diversity index and the number of catabolized substrates in Biolog plate (the R index) were much higher in the group subjected to mixed mycotoxicosis.

Topics: Animals; Bacteria, Aerobic; Bacterial Load; Biodiversity; Clostridium; Colon, Ascending; Colony Count, Microbial; Enterococcaceae; Escherichia coli; Fungi; Fusarium; Microbiota; Mycotoxicosis; No-Observed-Adverse-Effect Level; Swine; Trichothecenes; Zearalenone

Members of the Fusarium graminearum species complex (FGSC) are important pathogens on wheat, maize, barley, and rice in China. Harvested grains are often contaminated by mycotoxins, such as the trichothecene nivalenol (NIV) and deoxynivalenol (DON) and the estrogenic mycotoxin zearalenone (ZEN), which is a big threat to humans and animals. In this study, 97 isolates were collected from maize, wheat, and rice in Jiangsu and Anhui provinces in 2013 and characterized by species- and chemotype-specific PCR. F. graminearum sensu stricto (s. str.) was predominant on maize, while most of the isolates collected from rice and wheat were identified as F. asiaticum. Fusarium isolates from three hosts varied in trichothecene chemotypes. The 3-acetyldeoxynivalenol (3ADON) chemotype predominated on wheat and rice population, while 15ADON was prevailing in the remaining isolates. Sequence analysis of the translation elongation factor 1α and trichodiene synthase indicated the accuracy of the above conclusion. Additionally, phylogenetic analysis suggested four groups with strong correlation with species, chemotype, and host. These isolates were also evaluated for their sensitivity to carbendazim and mycotoxins production. The maize population was less sensitive than the other two. The DON levels were similar in three populations, while those isolates on maize produced more ZEN. More DON was produced in carbendazim resistant strains than sensitive ones, but it seemed that carbendazim resistance had no effect on ZEN production in wheat culture.

Topics: Benzimidazoles; Carbamates; China; DNA, Fungal; Fungicides, Industrial; Fusarium; Genotype; Oryza; Phylogeny; Sequence Analysis, DNA; Trichothecenes; Triticum; Zea mays; Zearalenone

The objective of this study was to develop a 1-step simultaneous lateral flow strip test for the rapid and simple detection of deoxynivalenol (DON) and zearalenone (ZEA) in grains. Two monoclonal antibodies (MAbs) against DON and ZEA were respectively conjugated with gold nanoparticles and used to develop a lateral flow strip test for a single toxin and multiple toxins. First, individual lateral flow strips for a single toxin were optimized, and their conditions were used to develop a simultaneous lateral flow strip for multiple toxins. Limits of detection of both lateral flow strip tests for DON and ZEA were the same (DON: 50 ng/mL, ZEA: 1 ng/mL). Both methods showed cross-reactivity for α-zearalenol and β-zearalenol, but no cross-reaction to other mycotoxins. The results can be completed obtained within 15 min. The cut-off values of the simultaneous lateral flow strip for the spiked rice and corn were 500 and 10 ng/g for DON and ZEA, respectively. The results demonstrated that the developed simultaneous lateral flow strip test offers a rapid, easy-to-use, and portable analytical system and can be used as a convenient qualitative tool for the on-site detection of DON and ZEA in food and agricultural commodities.. Simultaneous lateral strip test is useful for a rapid detection of DON and ZEA at a time in food and grain samples.

Topics: Antibodies, Monoclonal; Edible Grain; Food Analysis; Food Contamination; Gold; Metal Nanoparticles; Reagent Strips; Sensitivity and Specificity; Time Factors; Trichothecenes; Zearalenone

In this study, the exposure of a German population (n = 101) to mycotoxins was assessed using an LC-MS/MS urinary multibiomarker approach. Food consumption of the participants was documented with a food frequency questionnaire to correlate mycotoxin exposure with individual nutritional habits.. The presence of 23 urinary biomarkers including trichothecenes (deoxynivalenol (DON), DON-3-glucuronide (DON-3-GlcA), T-2 toxin, HT-2 toxin (HT-2, HT-2-toxin-4-glucuronide (HT-2-GlcA), fumonisins (fumonisin B1, fumonisin B2), aflatoxins (aflatoxin B1, aflatoxin G2, aflatoxin B2, aflatoxin M1), zearalenone and derivatives (zearalanone, α-zearalenol, β-zearalenol, zearalenone-14-O-glucuronide, zearalanone-14-O-glucuronide, α-zearalenol-14-O-glucuronide/β-zearalenol-14-O-glucuronide), ochratoxin A, ochratoxin alpha, enniatin B and dihydrocitrinone was evaluated using a validated, sensitive "dilute and shoot"-LC-MS/MS method applying Scheduled MRM(TM) technology. Six mycotoxins and urinary metabolites were detected (DON, DON-3-GlcA, zearalenone-14-O-glucuronide, T-2 toxin, enniatin B, and dihydrocitrinone) in 87% of the samples in single- or co-occurence. Only DON and DON-3-GlcA were detectable in quantifiable amounts. A provisional mean daily intake of 0.52 μg DON/kg body weight was calculated. No statistical evidence for the correlation of staple food intake and urinary biomarker concentration could be determined.. The results of this study suggest a low everyday exposure of the investigated German population to mycotoxins, but reveal peak exposures above the widely accepted tolerable daily intake to DON in parts of the population.

Topics: Adult; Aflatoxins; Chromatography, High Pressure Liquid; Chromatography, Liquid; Feeding Behavior; Female; Food Contamination; Food Microbiology; Fumonisins; Germany; Glucuronides; Humans; Male; Mycotoxins; Ochratoxins; Reproducibility of Results; Surveys and Questionnaires; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Young Adult; Zearalenone; Zeranol

A survey of the contamination of wheat, barley, and Japanese retail food by four Fusarium mycotoxins, deoxynivalenol (DON), zearalenone (ZEN), T-2 toxin (T-2), and HT-2 toxin (HT-2), was performed between 2010 and 2012. A method for the simultaneous determination of the four mycotoxins by liquid chromatography-tandem mass spectrometry was validated by a small-scale interlaboratory study using two spiked wheat samples (DON was spiked at 20 and 100 μg/kg and ZEN, T-2, and HT-2 at 6 and 20 μg/kg in the respective samples). The recovery of the four mycotoxins ranged from 77.3 to 107.2%. A total of 557 samples of 10 different commodities were analyzed over 3 years by this validated method. Both T-2 and HT-2 were detected in wheat, wheat flour, barley, Job's tears products, beer, corn grits, azuki beans, soybeans, and rice with mixed grains. Only T-2 toxin was detected in sesame seeds. The highest concentrations of T-2 toxin (48.4 μg/kg) and HT-2 toxin (85.0 μg/kg) were present in azuki beans and wheat, respectively. DON was frequently detected in wheat, wheat flour, beer, and corn grits. The contamination level of wheat was below the provisional standard in Japan (1,100 μg/kg). The maximum contamination level of DON was present in a sample of a Job's tears product (1,093 μg/kg). ZEN was frequently detected in Job's tears products, corn grits, azuki beans, rice with mixed grains, and sesame seeds. A sample of a Job's tears product presented the highest ZEN contamination (153 μg/kg). These results indicate that continuous monitoring by multiple laboratories is effective and necessary due to the percentage of positive samples detected.

Topics: Beer; Flour; Food Contamination; Food Microbiology; Fusarium; Hordeum; Japan; Mycotoxins; T-2 Toxin; Trichothecenes; Triticum; Zearalenone

This study was performed to assess the individual and combined toxic effects of aflatoxin B1 (AFB1), zearalenone (ZEA) and deoxynivalenol (DON) within the liver of mice. A total of 56 4-week-old weanling female mice were divided into seven groups (n = 8). For 2 weeks, each group received an oral administration of either solvent (control), AFB1, ZEA, DON, AFB1 + ZEA, AFB1 + DON or ZEA + DON per day. The results showed that AFB1, ZEA and DON induced liver injury, indicated by elevated relative liver weight, activities of alanine aminotransferase (ALT) and/or aspartate aminotransferase (AST), as well as decreased albumin (ALB) and/or total protein (TP) concentration in the serum. These mycotoxins also decreased hepatic total antioxidant capacity (T-AOC), and/or increased the concentration of malondialdehyde (MDA). Moreover, AFB1 + DON displayed synergistic effects, while AFB1 + ZEA displayed antagonistic effects on those parameters previously described. Furthermore, the apoptotic potential was demonstrated associated with an upregulation of the apoptotic genes Caspase-3 and Bax, along with a downregulation of the antiapoptotic gene Bcl-2 in liver. In conclusion, this study provides a better understanding of the toxic effects of AFB1, ZEA, DON, alone or in combinations on the liver of mice, which could contribute to the risk assessment of these mycotoxins in food and feed.

Topics: Aflatoxin B1; Alanine Transaminase; Animals; Antioxidants; Apoptosis Regulatory Proteins; Aspartate Aminotransferases; Chemical and Drug Induced Liver Injury; Drug Interactions; Female; Growth; Liver Function Tests; Malondialdehyde; Mice; Mycotoxins; Organ Size; Serum Albumin; Trichothecenes; Zearalenone

To evaluate the effects of the mycotoxins deoxynivalenol (DON) and zearalenone (ZEA) on pigs and the benefits of two mycotoxin mitigation strategies, gilts (n = 84, 9.1 ± 0.1 kg) were allotted to four treatments: CON (control); MT (4.8 mg/kg feed DON and 0.3 mg/kg feed ZEA); MT-YC (MT + 2 g/kg of yeast cell wall product); and MT-YF (MT + 2 g/kg of yeast fermentation product). After 42 days of feeding, pigs fed MT had reduced (p < 0.05) growth performance compared with pigs fed CON. Pigs fed MT-YF had greater (p < 0.05) average daily gain and tended to have greater (p = 0.080) average daily feed intake than MT, whereas pigs fed MT-YC did not differ from MT. Oxidative DNA damage increased (p < 0.05) in MT, whereas pigs fed MT-YF tended to have lower (p = 0.067) oxidative stress. Liver hydropic degeneration was increased (p < 0.05) in MT in contrast to CON and MT-YF, and tended to be greater (p = 0.079) than MT-YC. Collectively, feeding diets contaminated with mycotoxins significantly reduced growth performance and impacted pig health. The yeast additives had varied ability to reduce mycotoxin effects on pig growth and health, but may still play a beneficial role in reducing the overall impacts of a mycotoxin challenge on pigs.

Topics: Animal Feed; Animals; Food Contamination; Food Microbiology; Liver; Oxidative Stress; Saccharomyces cerevisiae; Swine; Trichothecenes; Zearalenone

Fusarium toxin-contaminated ground maize was hydrothermally treated in the presence of different combinations of chemicals in order to simultaneously reduce zearalenone (ZEA) and deoxynivalenol (DON) concentrations. Treatments were carried out in a laboratory conditioner at 80 °C and 17 % moisture. Six different treatments were performed, consisting of 3 doses of methylamine (MMA; 2.5, 5 and 10 g/kg maize) at a constant dose of 5 g sodium metabisulfite (SBS)/kg, either with or without the addition of 20 g calcium hydroxide (Ca(OH)2)/kg. The used maize was contaminated with approximately 45.99 mg DON/kg and 3.46 mg ZEA/kg. Without the addition of Ca(OH)2, DON reductions reached approximately 82% after 1-min treatment and the toxin disappeared nearly completely after 10 min when 2.5 or 5 g MMA were applied. ZEA concentrations were only marginally affected. In the presence of Ca(OH)2, reductions in DON concentrations were lower, but were enhanced by increasing doses of MMA. ZEA concentrations were reduced by 72, 85 and 95% within the first 5 min of the treatment at MMA dosages of 2.5, 5 and 10 g/kg maize, respectively. The application of SBS in combination with a strong alkaline during hydrothermal treatment seems to be a promising approach to simultaneously decontaminate even high amounts of DON and ZEA in ground maize and may contribute to reduce the toxin load of diets.

Topics: Calcium Hydroxide; Decontamination; Food Contamination; Hot Temperature; Methylamines; Sulfites; Trichothecenes; Zea mays; Zearalenone

Mycotoxins which mainly consist of Aflatoxin (AF), Zearalenone (ZEN) and Deoxynivalenol (DON) are commonly found in many food commodities. Although each component has been shown to cause liver toxicity and oxidative stress in several species, there is no evidence regarding the effect of naturally contained multiple mycotoxins on tissue toxicity and oxidative stress in vivo. In the present study, mycotoxins-contaminated maize (AF 597 µg/kg, ZEN 729 µg/kg, DON 3.1 mg/kg maize) was incorporated into the diet at three different doses (0, 5 and 20%) to feed the mice, and blood and tissue samples were collected to examine the oxidative stress related indexes. The results showed that the indexes of liver, kidney and spleen were all increased and the liver and kidney morphologies changed in the mycotoxin-treated mice. Also, the treatment resulted in the elevated glutathione peroxidase (GPx) activity and malondialdehyde (MDA) level in the serum and liver, indicating the presence of the oxidative stress. Moreover, the decrease of catalase (CAT) activity in the serum, liver and kidney as well as superoxide dismutase (SOD) activity in the liver and kidney tissue further confirmed the occurrence of oxidative stress. In conclusion, our data indicate that the naturally contained mycotoxins are toxic in vivo and able to induce the oxidant stress in the mouse.

Topics: Aflatoxins; Alanine Transaminase; Animals; Aspartate Aminotransferases; Diet; Female; Kidney; Liver; Mice; Oxidative Stress; Spleen; Trichothecenes; Zea mays; Zearalenone

Fusarium mycotoxins occur worldwide in foods such as cereals and animal forages, leading to acute and chronic exposures in human and animals. Intestinal epithelial cells (IECs) are an important first target site for these dietary toxins. This study investigated the cytotoxicity of four common Fusarium mycotoxins, deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEA) and fumonisin B1 (FB1) on a normal porcine jejunal epithelial cell line, IPEC-J2. A dose response relationship between individual mycotoxins and cell viability (MTT assay) was initially investigated, and subsequently cytotoxic and non-cytotoxic concentrations were selected to investigate combinations of two, three and all four of the mycotoxins. For individual mycotoxins, a dose response was observed with cell viability, such that the potency ranking was NIV>DON>ZEA>FB1. At cytotoxic doses of individual mycotoxins, all mixtures gave reduced cell viability compared to control. At noncytotoxic concentrations of individual mycotoxins, all mixtures were cytotoxic with DON-NIV, DON-ZEA, DON-NIV-FB1, DON-ZEA-FB1, NIV-ZEA-FB1 and all four mixed causing the greatest loss of cell viability. The latter observation in particular raises concerns over safety margins based on single toxin species, and suggests that the effects of multiple complex mixtures need to be better understood to assess health risks.

Topics: Animals; Cell Survival; Cells, Cultured; Dose-Response Relationship, Drug; Drug Synergism; Epithelial Cells; Fumonisins; Jejunum; Mycotoxins; Swine; Trichothecenes; Zearalenone

We assessed the production of the mycotoxins fumonisin, deoxynivalenol and zearalenone during the ensiling of corn. Corn was harvested at yellow-ripe or full-ripe stage and separated into the stem and leaf parts and the ear parts, including bracts. Each material was ensiled under five conditions: (1) no fungus added, anaerobic conditions; (2) no fungus added, aerobic conditions; (3) mycotoxin-producing fungus added, anaerobic conditions; (4) mycotoxin-producing fungus added, aerobic conditions; and (5) mycotoxin-producing fungus added to autoclaved material, aerobic conditions. After 40 days of ensilage, we analyzed the silage fermentative quality and mycotoxin concentration. The fermentative quality of all materials was good in treatments (1) and (3), because the pH < 4 increased the lactic acid content preventing mycotoxin levels from increasing. In treatments (2) and (4), fermentative quality of all materials was poor, and mycotoxin levels were slightly increased. In treatment (5), fermentative quality was poor, and mycotoxin levels were increased remarkably. These results indicate that mycotoxins are not produced under anaerobic conditions and are hardly produced under aerobic condition during the ensiling of corn. Our findings suggest that almost all mycotoxins in corn silage are produced pre-harvest.

Topics: Fumonisins; Silage; Trichothecenes; Zea mays; Zearalenone

This study reports on the detailed investigation of human deoxynivalenol (DON) and zearalenone (ZEN) in vivo metabolism through the analysis of urine samples obtained from one volunteer following a naturally contaminated diet containing 138μg DON and 10μg ZEN over a period of four days. Based on the mycotoxin intake and the concentrations of mycotoxin conjugates in urine, a mass balance was established. The average rates of DON excretion and glucuronidation were determined to be 68 and 76%, respectively. The investigation of formed glucuronides revealed DON-15-glucuronide as main conjugation product besides DON-3-glucuronide. Furthermore, for the first time in human urine a third DON-glucuronide was detected and the fate of ingested masked DON forms (3-acetyl-DON and DON-3-glucoside) was preliminary assessed. The mean excretion rate of ZEN was determined to be 9.4%. ZEN was mainly present in its glucuronide form and in some samples ZEN-14-glucuronide was directly determined 3-10h after exposure. For the first time concrete figures have become available for the excretion pattern of DON and ZEN-glucuronides throughout a day, the comparison of total DON in 24h and first morning urine samples and the urinary excretion rate of total ZEN in humans following exposure through naturally contaminated food. Therefore, valuable preliminary information has been obtained through the chosen experimental approach although the study involved only one single individual and needs to be confirmed in larger monitoring studies. The presented experiment contributes to a better understanding of human DON and ZEN in vivo metabolism and thereby supports advanced exposure and risk assessment to increase food safety and examine the relationship between these mycotoxins and potentially associated chronic diseases in the future.

Topics: Adult; Chromatography, High Pressure Liquid; Food Contamination; Food Microbiology; Fusarium; Glucuronides; Humans; Male; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Time Factors; Trichothecenes; Zearalenone

In the present study, we evaluated the nephrotoxicity of individual mycotoxins and combinations of aflatoxin B1 (AFB1), zearalenone (ZEA), deoxynivalenol (DON), and fumonisin B1 (FB1) to livestock using porcine kidney 15 cells (PK-15) as a disease model via biochemical approaches. The toxicity of individual mycotoxins on cell viability and cell membrane damage was determined using the MTT and lactate dehydrogenase (LDH) assays, respectively. Individual cytotoxicity of mycotoxins in increasing order were FB1

Topics: Aflatoxin B1; Animals; Apoptosis; Cell Line; Cell Survival; Fumonisins; Kidney; Mycotoxins; Reactive Oxygen Species; Swine; Trichothecenes; Zearalenone

Deoxynivalenol (DON) and zearalenone (ZEN) contaminated maize was hydrothermally treated in the presence of sodium metabisulphite (SBS), methylamine and calcium hydroxide (Ca(OH)2) and included into diets for female piglets to evaluate effects on performance, organ weights, development of hyperestrogenism, serum biochemical parameters, stimulation of peripheral blood mononuclear cells and toxin residues in serum. For this purpose, both uncontaminated maize (CON) and Fusarium toxin-contaminated maize (FUS) were included into diets either untreated (-) or treated (+) according to a 2 by 2-factorial design. One-hundred female weaned piglets were assigned to one of the four treatment groups (n = 25) CON-, CON+, FUS- and FUS+ with DON/ZEN concentrations of 0.43/0.03, 0.04/0.0, 3.67/0.32 and 0.36/0.08 mg per kg diet, respectively. After a feeding period of 27 days, 20 piglets (n = 5) were slaughtered. Performance parameters such as feed intake, live weight gain and feed-to-gain ratio remained unaffected by the treatments. Uterus weights were significantly reduced in group FUS+ compared to FUS- (p = 0.028), while visceral organ weights were not influenced. Vulva width in relation to body weight was highest in group FUS- at the end of the trial, while hydrothermal treatment significantly reduced the parameter (p < 0.01). The highest toxin and toxin metabolite concentrations in serum were detected in group FUS-, whereas ingestion of diet FUS+ reduced the concentrations to the level of the control groups. Serum biochemical and haematological parameters were mainly within the given reference ranges and showed no treatment-related alterations. Stimulation of peripheral blood mononuclear cells was not affected. An effective detoxification of maize by hydrothermal treatment in the presence of SBS, methylamine and Ca(OH)2 could be demonstrated by means of serum toxin analyses. No undesired side effects of the treated-feed stuff or the chemicals themselves on the health of piglets were detected.

Topics: Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Calcium Hydroxide; Diet; Female; Food Contamination; Fusarium; Methylamines; Sulfites; Swine; Trichothecenes; Zea mays; Zearalenone

Fusarium graminearum is a necrotrophic plant pathogen of cereals that produces mycotoxins such as deoxynivalenol (DON) and zearalenone (ZEA) in grains. The stress-activated mitogen-activated protein kinase FgOS-2 is a central regulator in F. graminearum and controls, among others, virulence and DON and ZEA production. Here, we characterized the ATF/CREB-activating transcription factor FgAtf1, a regulator that functions downstream of FgOS-2. We created deletion and overexpression mutants of Fgatf1, the latter being also in an FgOS-2 deletion mutant. FgAtf1 localizes to the nucleus and appears to interact with FgOS-2 under osmotic stress conditions. Deletion mutants in Fgatf1 (ΔFgatf1) are more sensitive to osmotic stress and less sensitive to oxidative stress compared with the wild type. Furthermore, sexual reproduction is delayed. ΔFgatf1 strains produced higher amounts of DON under in vitro induction conditions than that of the wild type. However, during wheat infection, DON production by ΔFgatf1 is strongly reduced. The ΔFgatf1 strains displayed strongly reduced virulence to wheat and maize. Interestingly, constitutive expression of Fgatf1 in the wild type led to hypervirulence on wheat, maize, and Brachypodium distachyon. Moreover, constitutive expression of Fgatf1 in the ΔFgOS-2 mutant background almost complements ΔFgOS-2-phenotypes. These data suggest that FgAtf1 may be the most important transcription factor regulated by FgOS-2.

Topics: Activating Transcription Factor 1; Adaptation, Physiological; Brachypodium; Cell Nucleus; Edible Grain; Fungal Proteins; Fusarium; Gene Expression; Gene Expression Regulation, Fungal; Inflorescence; Mitogen-Activated Protein Kinases; Models, Biological; Osmotic Pressure; Oxidative Stress; Plant Diseases; Secondary Metabolism; Sequence Deletion; Spores, Fungal; Trichothecenes; Triticum; Virulence; Zea mays; Zearalenone

Wheat is often infected by Fusarium species producing mycotoxins, which may pose health risks to humans and animals. Deoxynivalenol (DON) is the most important Fusarium toxin in Swedish wheat and has previously been shown to be produced mainly by Fusarium graminearum. However, less is known about the co-occurrence of DON and F. graminearum with other toxins and Fusarium species in Sweden. This study examined the distribution of the most important toxigenic Fusarium species and their toxins in winter wheat (2009 and 2011) and spring wheat (2010 and 2011). DNA from seven species was quantified with qPCR and the toxin levels were quantified with a multitoxin analysis method based on liquid chromatography/electrospray ionisation-tandem mass spectrometry (HPLC/ESI-MS/MS). The method enabled detection of many fungal metabolites, including DON, zearalenone (ZEA), nivalenol (NIV), T-2 toxin, HT-2 toxins, moniliformin (MON), beauvericin (BEA), and enniatins (ENNs). It was found that Fusarium poae and Fusarium avenaceum were present in almost all samples. Other common Fusarium species were F. graminearum and F. culmorum, present in more than 70% of samples. Several species occurred at lower DNA levels in 2011 than in other years, but the reverse was true for F. graminearum and Fusarium langsethiae. The most prevalent toxins were ENNs, present in 100% of samples. DON was also common, especially in spring wheat, whereas ZEA and NIV were common in 2009 and in winter wheat, but less common in 2011 and in spring wheat. Only three samples of spring wheat contained T-2 or HT-2 above LOQ. Annual mean levels of several mycotoxins were significantly lower in 2011 than in other years, but the reverse applied for DON. The strongest correlations between mycotoxin and Fusarium DNA levels were found between F. avenaceum and ENNs (r(2) = 0.67) and MON (r(2) = 0.62), and F. graminearum and DON (r(2) = 0.74). These results show that several Fusarium species and toxins co-occur in wheat. The highest toxin levels were detected in spring wheat and DON and ENNs, the latter belonging to the group of so called "emerging toxins", which were the most prevalent toxins and those occurring at the highest levels.

Topics: Chromatography, High Pressure Liquid; Cyclobutanes; Depsipeptides; DNA, Fungal; Food Contamination; Fusarium; Real-Time Polymerase Chain Reaction; Sweden; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Triticum; Zearalenone

Fusarium moulds frequently contaminate oats and other cereals world-wide, including those grown in Northern Europe. To investigate the presence of toxigenic Fusarium species and their toxins in oats, samples were taken during 2010 and 2011 in three geographical regions of Sweden (east, west, south). The samples were analysed by real-time PCR for the specific infection level of seven Fusarium species associated with oats and other cereals (Fusarium poae, Fusarium graminearum, Fusarium langsethiae, Fusarium culmorum, Fusarium tricinctum, Fusarium sporotrichioides and Fusarium avenaceum) and with a multi-mycotoxin method based on liquid chromatography/electrospray ionisation-tandem mass spectrometry (HPLC/ESI-MS/MS) for the detection of many fungal metabolites, including deoxynivalenol (DON), zearalenone (ZEA), nivalenol (NIV), T-2 toxin, HT-2 toxins, moniliformin (MON), beauvericin (BEA) and enniatins (ENNs). Most samples contained at least four of the seven Fusarium species analysed and F. poae, F. langsethiae and F. avenaceum were present in approximately 90-100% of all samples. The most common toxins detected were DON, NIV, BEA and ENNs, which were present in more than 90% of samples. Most Fusarium species and their toxins occurred in higher concentrations in 2010 than in 2011, with the exception of DON and its main producer F. graminearum. Significant regional differences were detected for some moulds and mycotoxins, with higher levels of F. graminearum, DON and ZEA in western Sweden than in the east (P<0.05) and higher levels of F. tricinctum and MON in the south (P<0.05). Correlation analysis showed significant correlations between many Fusarium species and toxin levels. For example, F. tricinctum was significantly correlated to F. avenaceum (r = 0.72, P<0.001), DON to ZEA (r = 0.52, P<0.001), DON to F. graminearum (r = 0.77, P<0.001) and the sum of T-2 and HT-2 to F. langsethiae (r = 0.77, P<0.001). The multi-toxin approach employed allowed simultaneous detection of many Fusarium mycotoxins in each sample. In combination with real-time PCR analysis of seven toxigenic Fusarium spp., the results gave an overall picture of the presence of Fusarium and their toxins in Swedish oats and revealed significant annual and regional differences. This is the first study of the so-called emerging mycotoxins (e.g., ENNs, MON and BEA) in oats grown in Sweden.

Topics: Avena; Chromatography, High Pressure Liquid; Cyclobutanes; Depsipeptides; DNA, Fungal; Edible Grain; Food Contamination; Fusarium; Geography; Real-Time Polymerase Chain Reaction; Sweden; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

Subsistence farmers are exposed to a range of mycotoxins. This study applied novel urinary multi-mycotoxin LC-MS/MS methods to determine multiple exposure biomarkers in the high oesophageal cancer region, Transkei, South Africa. Fifty-three female participants donated part of their maize-based evening meal and first void morning urine, which was analysed both with sample clean-up (single and multi-biomarker) and by a 'dilute-and-shoot' multi-biomarker method. Results were corrected for recovery with LOD for not detected. A single biomarker method detected fumonisin B1 (FB1) (87% incidence; mean±standard deviation 0.342±0.466 ng/mg creatinine) and deoxynivalenol (100%; mean 20.4±49.4 ng/mg creatinine) after hydrolysis with β-glucuronidase. The multi-biomarker 'dilute-and-shoot' method indicated deoxynivalenol-15-glucuronide was predominantly present. A multi-biomarker method with β-glucuronidase and immunoaffinity clean-up determined zearalenone (100%; 0.529±1.60 ng/mg creatinine), FB1 (96%; 1.52±2.17 ng/mg creatinine), α-zearalenol (92%; 0.614±1.91 ng/mg creatinine), deoxynivalenol (87%; 11.3±27.1 ng/mg creatinine), β-zearalenol (75%; 0.702±2.95 ng/mg creatinine) and ochratoxin A (98%; 0.041±0.086 ng/mg creatinine). These demonstrate the value of multi-biomarker methods in measuring exposures in populations exposed to multiple mycotoxins. This is the first finding of urinary deoxynivalenol, zearalenone, their conjugates, ochratoxin A and zearalenols in Transkei.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers; Environmental Exposure; Farmers; Female; Food Contamination; Fumonisins; Humans; Middle Aged; Mycotoxins; Ochratoxins; Rural Population; South Africa; Tandem Mass Spectrometry; Trichothecenes; Young Adult; Zea mays; Zearalenone; Zeranol

Analytical methods were validated for the analysis of fumonisins (FB1 and FB2), deoxynivalenol (DON) and zearalenone (ZEA) in maize germ, corn oil and margarine. A survey of 74 samples consisting of 12 wet-milled maize germ, 12 dry-milled maize germ, 25 refined corn oil, and 25 corn oil margarine was conducted. Results revealed that 100% and 87.5% of maize germ samples presented FB1 and FB2, respectively, attaining concentrations for the sum of both toxins of 1302±541 μg kg(-1) in wet-milled and 820±831 μg kg(-1) in dry-milled maize germ. The lower incidence of FB1, FB2 and DON in edible oil and margarine (4-8%) may be related with the industrial processes for their obtaining besides the high water-solubility of these mycotoxins. In contrast, 25% of maize germ samples were positive for ZEA as well as 32% of corn oil and 24% of margarine, which may be related with its lipophilic nature. A number of samples exceeded the maximum limits indicating that strict control is needed, though estimated dietary exposure was less than 0.2% tolerable daily intakes in all cases.

Topics: Corn Oil; Food Contamination; Fumonisins; Fusarium; Margarine; Mycotoxins; Reproducibility of Results; Trichothecenes; Zea mays; Zearalenone

Bio-monitoring of human exposure to mycotoxin has mostly been limited to a few individually measured mycotoxin biomarkers. This study aimed to determine the frequency and level of exposure to multiple mycotoxins in human urine from Cameroonian adults. 175 Urine samples (83% from HIV-positive individuals) and food frequency questionnaire responses were collected from consenting Cameroonians, and analyzed for 15 mycotoxins and relevant metabolites using LC-ESI-MS/MS. In total, eleven analytes were detected individually or in combinations in 110/175 (63%) samples including the biomarkers aflatoxin M1, fumonisin B1, ochratoxin A and total deoxynivalenol. Additionally, important mycotoxins and metabolites thereof, such as fumonisin B2, nivalenol and zearalenone, were determined, some for the first time in urine following dietary exposures. Multi-mycotoxin contamination was common with one HIV-positive individual exposed to five mycotoxins, a severe case of co-exposure that has never been reported in adults before. For the first time in Africa or elsewhere, this study quantified eleven mycotoxin biomarkers and bio-measures in urine from adults. For several mycotoxins estimates indicate that the tolerable daily intake is being exceeded in this study population. Given that many mycotoxins adversely affect the immune system, future studies will examine whether combinations of mycotoxins negatively impact Cameroonian population particularly immune-suppressed individuals.

Topics: Adolescent; Adult; Biomarkers; Cameroon; Environmental Exposure; Environmental Monitoring; Feeding Behavior; Female; Food Contamination; Fumonisins; Glucuronides; Humans; Male; Middle Aged; Mycotoxins; No-Observed-Adverse-Effect Level; Ochratoxins; Surveys and Questionnaires; Trichothecenes; Young Adult; Zearalenone

The aim of the study was to verify the hypothesis that intoxication with low doses of mycotoxins leads to changes in the mRNA expression levels of nitric oxide synthase-1 and nitric oxide synthase-2 genes in tissues of the gastrointestinal tract and the liver. The experiment involved four groups of immature gilts (with body weight of up to 25 kg) which were orally administered zearalenone in a daily dose of 40 μg/kg BW (group Z, n = 18), deoxynivalenol at 12 μg/kg BW (group D, n = 18), zearalenone and deoxynivalenol (group M, n = 18) or placebo (group C, n = 21) over a period of 42 days. The lowest mRNA expression levels of nitric oxide synthase-1 and nitric oxide synthase-2 genes were noted in the sixth week of the study, in particular in group M. Our results suggest that the presence of low mycotoxin doses in feed slows down the mRNA expression of both nitric oxide synthase isomers, which probably lowers the concentrations of nitric oxide, a common precursor of inflammation.

Topics: Animal Feed; Animals; Dose-Response Relationship, Drug; Gastrointestinal Tract; Gene Expression Regulation; Liver; Mycotoxicosis; Nitric Oxide; Nitric Oxide Synthase Type II; Nitric Oxide Synthase Type III; RNA, Messenger; Swine; Trichothecenes; Zearalenone

The objective of this study was to evaluate the occurrence and levels of deoxynivalenol (DON), fumonisins B1 and B2 (FBs), and zearalenone (ZEN) contaminants in animal feeds used in Korea in 2012. Contamination with DON was observed in 91.33% and 53.33% in compound feeds and feed ingredients, respectively. Among compound feeds, poultry layer feed (laying) exhibited the highest contaminant level of 1.492 mg/kg. FBs contaminants were present in compound feeds and feed ingredients at 93.33% and 83.33%, respectively. Most poultry broiler (early) feeds were highly contaminated with FBs, and one of these feeds detected the level as 12.823 mg/kg as the highest level. The levels of ZEN in compound feeds and feed ingredients were 71.33% and 47%, respectively. Ninety-eight percent of compound feeds for cattle were contaminated with ZEN, and the highest contamination level of 0.405 mg/kg was observed in cattle fatting feeds.

Topics: Animal Feed; Animals; Cattle; Chromatography, Liquid; Food Contamination; Food Microbiology; Fumonisins; Limit of Detection; Mycotoxins; Poultry; Republic of Korea; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

Monitoring results of food grain contamination with fusariotoxins-deoxynivalenol (DON), zearalenone (ZEN), fumonisins (FB1&FB2), T-2 and HT-2 toxins-are presented. Harvests of 2005-2010 in different regions of Russia were investigated. The occurrence of DON in wheat was 8%, barley 9%, oats 4%, rye 2% and maize 2%. The highest frequency of ZEN contamination was found in oats, the lowest in wheat. Calculated average daily intake of DON varied from 0.066 to 0.096 µg/kg body weight, the highest being found in the Southern region, but substantially lower than the provisional maximum tolerable daily intake. The results of enzyme-linked immunosorbent assay and high-performance liquid chromatography-mass spectrometry analysis demonstrated the presence of T-2 toxin in 14% and HT-2 toxin in 17% of all samples. The maximum level of T-2 toxin was exceeded only in one sample of barley. Relatively high frequency and levels of FB1&FB2 contamination were found in maize.

Topics: Crops, Agricultural; Diet; Edible Grain; Food Contamination; Food Inspection; Fumonisins; Fusarium; Guideline Adherence; Health Policy; Health Promotion; Humans; Mycotoxins; Poisons; Russia; Seeds; Spatio-Temporal Analysis; T-2 Toxin; Trichothecenes; Zearalenone

The persistence of mycotoxins and their metabolites in agricultural products is a major safety concern because of their high resistance to all kinds of decontamination techniques. In this study, we evaluated the effectiveness of the pulsed light technology for the degradation of mycotoxins. We report that eight flashes of pulsed light destroyed of 84.5 ± 1.9, 72.5 ± 1.1, 92.7 ± 0.8 and 98.1 ± 0.2% of zearalenone, deoxynivalenol, aflatoxin B1 and ochratoxin in solution. The degradation of the molecules was monitored by HPLC and LC-MS/MS analysis. We estimated the potential toxicity of zearalenone and deoxynivelenol after exposure to a pulsed light treatment using the Caenorhabditis elegans survival tests. The genotoxicity of aflatoxin B1 was also investigated using a complete Ames test. The results show that the treatment of zearalenone and deoxynivelenol by single or multiple flashes of pulsed light is associated with a stagnation or marginal decrease of the toxicity of the mycotoxins and that treatment of aflatoxin B1 by pulsed light can completely eliminate the mutagenic potential of this mycotoxin. This work provides the first demonstration of a nonthermal technology allowing mycotoxin destruction and inactivation of their mutagenic activity.

Topics: Aflatoxin B1; Chromatography, High Pressure Liquid; Chromatography, Liquid; Ochratoxins; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

Mycotoxins lead to economic losses in animal production. A way to counteract mycotoxicosis is the use of detoxifiers. The European Food Safety Authority stated that the efficacy of detoxifiers should be investigated based on toxicokinetic studies. Little information is available on the absolute oral bioavailability and the toxicokinetic parameters of deoxynivalenol, T-2 and zearalenone in broilers. Toxins were administered intravenously and orally in a two-way cross-over design. For deoxynivalenol a bolus of 0.75mg/kg BW was administered, for T-2 toxin 0.02mg/kg BW and for zearalenone 0.3mg/kg BW. Blood was collected at several time points. Plasma levels of the mycotoxins and their metabolite(s) were quantified using LC-MS/MS methods and toxicokinetic parameters were analyzed. Deoxynivalenol has a low absolute oral bioavailability (19.3%). For zearalenone and T-2 no plasma levels above the limit of quantification were observed after an oral bolus. Volumes of distribution were recorded, i.e. 4.99, 0.14 and 22.26L/kg for deoxynivalenol, T-2 toxin and zearalenone, respectively. Total body clearance was 0.12, 0.03 and 0.48L/minkg for deoxynivalenol, T-2 toxin and zearalenone, respectively. After IV administration, T-2 toxin had the shortest elimination half-life (3.9min), followed by deoxynivalenol (27.9min) and zearalenone (31.8min).

Topics: Administration, Oral; Animal Feed; Animals; Biological Availability; Chickens; Chromatography, Liquid; Half-Life; Injections, Intravenous; Pharmacokinetics; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

Direct determination of urinary mycotoxins is a better approach to assess individual's exposure than the indirect estimation from average dietary intakes. In this study, a new analytical method was developed and validated for simultaneous analysis of aflatoxin B1, deoxynivalenol, fumonisin B1, ochratoxin A, zearalenone and T2 toxin and their metabolites in pig urine. In total 12 analytes were selected. A salting-out assisted liquid-liquid extraction procedure was used for sample preparation. High performance liquid chromatography/tandem mass spectrometry was used for the separation and detection of all the analytes. The extraction recoveries were in a range of 70-108%, with the intra-day relative standard deviation and inter-day relative standard deviation lower than 25% for most of the compounds at 3 different concentration levels. Meanwhile the method bias for all the analytes did not exceed 20%. The limits of quantification ranged from 0.07ngmL(-1) for ochratoxin A to 3.3ngmL(-1) for deoxynivalenol. Matrix effect was evaluated in this study and matrix-matched calibration was used for quantification. The developed method was also validated for human urine as an extension of its application. Finally, the developed method was applied in a pilot study to analyze 28 pig urine samples. Deoxynivalenol, aflatoxin B1, fumonisin B1 and ochratoxin A were detected in these samples.

Topics: Aflatoxin B1; Animals; Chromatography, High Pressure Liquid; Fumonisins; Limit of Detection; Liquid-Liquid Extraction; Mycotoxins; Ochratoxins; Sus scrofa; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

The aflatoxins (AFs), deoxynivalenol (DON), ochratoxin A (OTA) and zearalenone (ZEA) are mycotoxins produced by fungal species which can contaminate, alone or simultaneously, cereal-based raw materials. Usually, the higher mycotoxins concentrations in cereals are found in the external layers of the grain (bran). Nowadays bran is increasingly consumed for its high fibre concentration. The objectives of this study were determining the concentration of these mycotoxins in bran samples intended for direct human consumption and to study the influence of some characteristics of the samples that may affect the mycotoxins content, there are not studies about fibre for direct human consumption. 67 bran samples from shops and supermarkets from two different Spanish cities were analyzed, being 37 samples of wheat bran and the remaining of oat bran. The results showed a major presence of DON in the analyzed samples, with levels above the EU legislation in some samples. Presence of DON was more frequent in wheat samples, compared to oats ones (p<0.05). Extruded or toasted samples, subjected to a heat treatment during processing, presented a significantly lower concentration of OTA, and differences between the organically and conventionally produced samples were also detected in OTA, which showed higher levels in the organic samples. Co-occurrence was frequently found between the Fusarium mycotoxins (ZEA and DON). Due to the high levels of DON in the analyzed samples, a calculation of DON intake has been made and it has been demonstrated that bran can account for an important percentage of DON exposure in the total diet.

Topics: Aflatoxins; Avena; Consumer Product Safety; Diet; Dietary Fiber; Dietary Supplements; Environmental Exposure; Food Contamination; Food Microbiology; Ochratoxins; Reproducibility of Results; Spain; Trichothecenes; Triticum; Zearalenone

The control of mycotoxins is a global challenge not only in human consumption but also in nutrition of farm animals including aquatic species. Fusarium toxins, such as deoxynivalenol (DON) and zearalenone (ZEN), are common contaminants of animal feed but no study reported the occurrence of both mycotoxins in fish feed so far. Here, we report for the first time the occurrence of DON and ZEN in samples of commercial fish feed designed for nutrition of cyprinids collected from central Europe. A maximal DON concentration of 825 μg kg(-1) feed was found in one feed whereas average values of 289 μg kg(-1) feed were noted. ZEN was the more prevalent mycotoxin but the concentrations were lower showing an average level of 67.9 μg kg(-1) feed.

Topics: Animal Feed; Animals; Aquaculture; Cyprinidae; Food Contamination; Food Microbiology; Trichothecenes; Zearalenone

The effect of Saccharomyces cerevisiae RC008 and RC016 strains, previously selected based on their aflatoxin B₁ mycotoxin binding ability and beneficial properties, against Aspergillus carbonarius and Fusarium graminearum under different interacting environmental conditions was evaluated. In vitro studies on the lag phase, growth rate and ochratoxin A/zearalenone and DON production were carried out under different regimens of a(w) (0.95 and 0.99); pH (4 and 6); temperature (25 and 37 °C) and oxygen availability (normal and reduced). Both yeast strains showed antagonistic activity and decreasing growth rate compared to the control. In general, the RC016 strain showed the greatest inhibitory activity. Except at the interacting condition 0.95 a(W), normal oxygen availability and 37 °C, at both pH values, A. carbonarius and F. graminearum were able to produce large amounts of mycotoxins in vitro. In general, a significant decrease in levels of mycotoxins in comparison with the control was observed. S. cerevisiae RC008 and RC016 could be considered as effective agents to reduce growth and OTA, ZEA and DON production at different interacting environmental conditions, related to those found in stored feedstuff. The beneficial and biocontrol properties of these strains are important in their use as novel additives for the control of mycotoxigenic fungi in stored feedstuffs.

Topics: Aflatoxin B1; Antibiosis; Aspergillus; Fusarium; Hydrogen-Ion Concentration; Mycotoxins; Ochratoxins; Oxygen; Saccharomyces cerevisiae; Temperature; Trichothecenes; Water; Zearalenone

Fusarium mycotoxins are secondary metabolites produced by Fusarium spp. in cereals. Among them, deoxynivalenol (DON) and zearalenone (ZEN) are widespread worldwide contaminants of cereal commodities and are ranked as the most important chronic dietary risk factors. Their conjugates, known as masked mycotoxins, have been described but are still not accounted for in risk assessment studies. This study demonstrates for the first time that DON and ZEN are effectively deconjugated by the human colonic microbiota, releasing their toxic aglycones and generating yet unidentified catabolites. For this reason, masked mycotoxins should be considered when evaluating population exposure.

Topics: Food Microbiology; Fusarium; Humans; Hydrolysis; Intestines; Metagenome; Mycotoxins; Trichothecenes; Zearalenone

An isolated occurrence of Fusarium head blight (FHB) of wheat was detected in the south-west region of Western Australia during the 2003 harvest season. The molecular identity of 23 isolates of Fusarium spp. collected from this region during the FHB outbreak confirmed the associated pathogens to be F. graminearum, F. acuminatum or F. tricinctum. Moreover, the toxicity of their crude extracts from Czapek-Dox liquid broth and millet seed cultures to brine shrimp (Artemia franciscana) was associated with high mortality levels. The main mycotoxins detected were type B trichothecenes (deoxynivalenol and 3-acetyldeoxynivalenol), enniatins, chlamydosporol and zearalenone. This study is the first report on the mycotoxin profiles of Fusarium spp. associated with FHB of wheat in Western Australia. This study highlights the need for monitoring not just for the presence of the specific Fusarium spp. present in any affected grain but also for their potential mycotoxin and other toxic secondary metabolites.

Topics: Animals; Artemisia; Chromatography, High Pressure Liquid; Depsipeptides; Edible Grain; Fusarium; Mycotoxins; Plant Diseases; Pyrones; Trichothecenes; Triticum; Western Australia; Zearalenone

The aim of this study was to investigate the contamination of pig feed with moulds and the occurrence of mycotoxins. A total of 30 feed samples were collected at different animal feed factories in the north-western part of Croatia. Mycological analysis showed that the total number of moulds ranged from 1 × 10(3) to 1 × 10(5) cfu/g with samples contaminated with Aspergillus spp. (63 %), Penicillium spp. (80 %), and Fusarium spp. (77 %). A determination of aflatoxin B1 (AFB1), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), T-2 toxin (T-2) and fumonisin (FUM) concentration was done using the validated ELISA method. The mean concentrations of AFB1 (0.5 ± 0.6 μg/kg), OTA (1.53 ± 0.42 μg/kg) and FUM (405 ± 298 μg/kg) were below the maximum levels or recommended values in the EU in all the investigated samples. The observed results indicated an increased contamination of pig feed with Fusarium mycotoxins DON and ZEA with mean concentrations of 817 ± 447 and 184 ± 214 μg/kg, higher than recommended in 40 and 17 % of the analysed samples, respectively.

Topics: Animal Feed; Animals; Aspergillus; Croatia; Food Contamination; Fungi; Fusarium; Mycotoxins; Penicillium; Trichothecenes; Zearalenone

Distiller's dried grains with solubles (DDGS) is a major coproduct of the fuel-ethanol industry and is becoming a popular low-cost ingredient for animal feed. Uncertainties regarding the risk factors in DDGS, such as level of mycotoxins, could limit its application in the animal feed industry. To provide a scientifically sound assessment of the prevalence and levels of mycotoxins in U.S. DDGS, we measured aflatoxins, deoxynivalenol, fumonisins, T-2 toxin, and zearalenone in 67 DDGS samples collected from 8 ethanol plants in the midwestern United States from 2009 to 2011. Among the five mycotoxins, deoxynivalenol was the main focus of the study because the crop year of 2009 was favorable for deoxynivalenol occurrence in corn. We learned that no more than 12% of the samples contained deoxynivalenol levels higher than the minimum advisory level for use in animal feed provided by the U.S. FDA, and the deoxynivalenol levels in all DDGS collected in 2011 were <2 mg/kg. Besides, intensive study showed that the enrichment of deoxynivalenol from contaminated corn to DDGS was about 3.5 times. With regard to the other mycotoxins in DDGS, the study suggested that (1) almost none of the DDGS samples produced in 2010 contained detectable aflatoxins and the highest level of aflatoxins in DDGS was 5.7 μg/kg; (2) no more than 6% of the samples contained fumonisin levels higher than the guidance level for feeding equids and rabbits provided by the U.S. FDA; (3) none of the samples contained T-2 higher than the detection limit; (4) most samples contained zearalenone levels between 100 and 300 μg/kg. This study was based on representative DDGS samples from the U.S. ethanol industry, and the data were collected using state-of-the-art analytical methodology. This study provided a comprehensive and scientifically sound assessment of the occurrence and levels of mycotoxins in DDGS produced from 2009 to early 2011 by the U.S. ethanol industry.

Topics: Aflatoxins; Animal Feed; Data Collection; Edible Grain; Ethanol; Food Contamination; Food Industry; Fumonisins; Midwestern United States; Mycotoxins; T-2 Toxin; Trichothecenes; Zea mays; Zearalenone

Contamination by mycotoxins is a major concern to the maize industry in north-east Italy where maize grain is often spoiled by Fusarium spp. In this work, fumonisins, deoxynivalenol and zearalenone were determined and an artificial neural network (ANN) model suitable for predicting mycotoxin contamination of maize at harvest time was developed.. The occurrence of deoxynivalenol and zearalenone was very limited, while fumonisins concentration ranged from 163 and to 3663 µg kg(-1) in 2007, and from 333 to 11473 µg kg(-1) in 2008. Statistical data analysis of factors affecting fumonisins concentration revealed that irrigation, chemical treatment against the European corn borer and harvest date significantly affected the level of contamination (P < 0.05), although the relevance of the factors was different in 2007 and 2008. The neural network approach showed a significant correlation between ascertained values and predictions based on agronomic data.. This is the first time that an artificial neural network has been used to predict fumonisin accumulation in maize: the prediction has been shown to have the potential for the development of a new approach for the rapid cataloging of grain lots.

Topics: Agriculture; Environment; Food Contamination; Fumonisins; Neural Networks, Computer; Seeds; Trichothecenes; Zea mays; Zearalenone

A total of 230 samples of processed rice and its sub-products or derived products were analysed to establish the co-occurrence of several mycotoxins. Samples were analysed in the period 2007-2009 due to the outbreak of beriberi associated with the consumption of rice stored in inappropriate conditions in Brazil. According to data from the Ministry of Health, 323 cases of disease were registered in 2006, of which at least 47 cases resulted in death. The occurrence of total aflatoxin (AFT) (aflatoxin B(1) + B(2) + G(1) + G(2)), ochratoxin A (OTA), zearalenone (ZON), deoxynivalenol (DON), and citreoviridin (CTV) was 58.7%, 40.0%, 45.2%, 8.3% and 22.5%, respectively. From 166 rice samples analysed, 55% had levels <0.11 µg kg(-1) for AFT. For OTA and ZON, of 165 rice samples analysed, 28% and 29% were contaminated with levels from 0.20 to 0.24 µg kg(-1) and from 3.6 to 290.0 µg kg(-1), respectively. One sample (0.6%) was contaminated with 4872.0 µg kg(-1) of ZON. A total of 91% of rice samples (n = 165) did not contain detectable DON (<30.00 µg kg(-1)), although the highest level of contamination was found to be 244 µg kg(-1). From the total of 65 samples analysed, 94% had no detectable CTV (<0.9 µg kg(-1)), with a range from 0.9 to 31.1 µg kg(-1) in 6% of the samples. The highest levels of contamination were found in rice sub-products or derived products from the husk and rice bran. Co-occurrence was observed for AFT and ZON in 17.0%, AFT and OTA in 24.2%, AFT and CTV in 6.2%, OTA and CTV in 4.6%, and ZON and CTV in 3.1%. These fractions were also the major contributors for the co-occurrence. The results found show the necessity of monitoring rice production.

Topics: Aflatoxin B1; Aflatoxins; Aurovertins; Brazil; Food Analysis; Food Contamination; Mycotoxins; Ochratoxins; Oryza; Reference Standards; Trichothecenes; Zearalenone

Mycotoxins are highly toxic contaminants in food, animal feed, and commodities. The study has developed an immunochip for quantifying the concentrations of six mycotoxins: aflatoxin B1, aflatoxin M1, deoxynivalenol, ochratoxin A, T-2 toxin, and zearalenone, which were added to drinking water. The complete antigens (Ags) of the mycotoxins were contact printed and immobilized onto agarose-modified glass slides with 12 physically isolated subarrays, based on the reaction of both diffusion and covalent bond. The optimal concentration of each antigen and antibody (Ab) was obtained using an Ag-Ab immunoassay. Based on the indirect competitive immunoassay for the simultaneous detection of six mycotoxins in one single chip, six standard curves with good logistic correlation (R(2)>0.97) were respectively plotted. The working ranges (0.04-1.69, 0.45-3.90, 20.20-69.23, 35.68-363.18, 0.11-1.81, and 0.08-7.47 ng/mL, respectively) were calculated, as well as the median inhibitory concentrations (0.31±0.04, 1.49±0.21, 34.54±1.30, 134.06±11.75, 0.49±0.05, and 1.54±0.22 ng/mL, respectively), when six mycotoxins were detected simultaneously. Finally, the recovery rates in drinking water generally ranged from 80% to 120% on the same chip, with an intra-assay coefficient of variation lower than 15%. We successfully established an immunochip for simultaneous detection of six mycotoxins within 4h, with advantages of using minimal samples and being visually semiquantitative with our naked eyes. In summary, the method could be developed on one single chip for detecting multiple contaminants in actual samples.

Topics: Aflatoxin B1; Aflatoxin M1; Antigens; Biosensing Techniques; Drinking Water; Food Contamination; Immunoassay; Ochratoxins; T-2 Toxin; Trichothecenes; Water Microbiology; Zearalenone

An LC-MS/MS method was developed and validated for the simultaneous determination of deoxynivalenol, zearalenone, T-2-toxin, HT-2-toxin and metabolites, including 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deoxynivalenol-3-glucoside, α-zearalenol, β-zearalenol, zearalenone-4-glucoside, α-zearalenol-4-glucoside, β-zearalenol-4-glucoside and zearalenone-4-sulfate in maize, wheat, oats, cornflakes and bread. Extraction was performed with acetonitrile/water/acetic acid (79/20/1, v/v/v) followed by a hexane defatting step. After filtration, the extract was evaporated and the residue was redissolved in mobile phase for injection. The mobile phase, which consisted of a mixture of methanol and water with 10 mM ammonium acetate, was adjusted to pH 3 with glacial acetic acid. A sample clean-up procedure was not included because of the low recoveries of free and masked mycotoxins and their differences in polarity. The method allowed the simultaneous determination of 13 Fusarium mycotoxins in a one-step chromatographic run using a Waters Acquity UPLC system coupled to a Quattro Premier XE mass spectrometer. The method was validated for several parameters such as linearity, apparent recovery, limit of detection, limit of quantification, precision, expanded measurement uncertainty and specificity. The limits of detection varied from 5 to 13 ng g⁻¹; those for the limit of quantification from 10 to 26 ng g⁻¹. The results of the performance characteristics of the developed LC-MS/MS method were in good agreement with the criteria mentioned in Commission Regulation (EC) No. 401/2006. Thirty samples of a variety of food and feed matrices were sampled and analysed between July 2010 and January 2011.

Topics: Animal Feed; Bread; Chromatography, High Pressure Liquid; Edible Grain; European Union; Food Contamination; Food Inspection; Fumonisins; Fusarium; Limit of Detection; Ochratoxins; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

Deoxynivalenol (DON) has been recently documented to deteriorate intestinal morphology in chickens at dietary doses that are regarded as safe for this species. The present trial was conducted to explore the significance of these morphological changes in relation to intestinal absorptive functionality and DON metabolism. Ross broilers at 7 d of age were fed either a basal diet (0.265 ± 0.048 mg of DON/kg; 0.013 ± 0.001 mg of zearalenone/kg), a low DON diet (1.68 mg of DON/kg; 0.145 ± 0.007 mg of zearalenone/kg), or a high DON diet (12.209 ± 1.149 mg of DON/kg; 1.094 ± 0.244 mg of zearalenone/kg). The DON diets (to variable degrees) progressively decreased the relative density (weight:length) of the small intestine with increasing exposure length, which could be correlated with a decrease in villus height in the small intestine. Short circuit current of the jejunal epithelium, reflecting transport function of the epithelium per unit area, was reduced (P = 0.001) in the birds fed the high DON diet. The increasing dietary level of DON linearly (P = 0.035) increased the length of the jejunum in wk 4 of exposure, resulting in conservation of macronutrient retention. Upon challenging the birds with a fixed amount of DON after wk 5 of exposure, higher (P ≤ 0.033) amounts of DON and the detoxification metabolite (de-epoxy-DON) were found at 5 h postchallenge in the guts of birds raised on the DON diets. The increasing level of previous exposure to DON linearly (P = 0.040) decreased the plasma level of DON in the birds at 1 h postchallenge. The amounts of zearalenone and its analogs in the gut and plasma also followed a trend similar to that for DON. These data suggest that intestines in chickens may adapt to a chronic DON challenge by morphological and functional modifications. The birds having previous exposure to Fusarium mycotoxins showed moderate detoxification coupled with reduced transfer of the mycotoxins to systemic circulation. Some metabolites of zearalenone found in this study were previously unknown for chickens.

Topics: Animal Feed; Animals; Chickens; Food Contamination; Fusariosis; Fusarium; Intestinal Diseases; Intestine, Small; Male; Mycotoxins; Organ Size; Poultry Diseases; Random Allocation; Stomach, Avian; Trichothecenes; Zearalenone

Fusarium graminearum and F. verticillioides are two very important mycotoxigenic species as they cause diverse diseases in crops. The effects of constant and cycling temperatures on growth and mycotoxin production of these species were studied on soybean based medium and on irradiated soya beans.. F. graminearum grew better when was incubated at 15, 20 and 15-20 °C (isothermal or cycling temperature) during 21 days of incubation. Maximum levels of zearalenone and deoxynivalenol (39.25 and 1040.4 µg g(-1), respectively) were detected on soya beans after 15 days of incubation and the optimal temperature for mycotoxin production was 15 °C for zearalenone and 20 °C for deoxynivalenol. F. verticillioides grew better at 25 °C in culture medium and at 15/20 °C and 15/25 °C on soybean seeds. Fumonisin B(1) was produced only in culture medium, and the maximum level (7.38 µg g(-1)) was found at 15 °C after 7 days of incubation.. When growth and mycotoxin production under cycling temperatures were predicted from the results under constant conditions, observed values were different from calculated for both species and substrate medium. Therefore, care should be taken if data at constant temperature conditions are to be extrapolated to real field conditions.

Topics: Food Microbiology; Fusarium; Glycine max; Mycotoxins; Seeds; Temperature; Trichothecenes; Zearalenone

Most recent information on the occurrence of Fusarium Head Blight species and related mycotoxins in wheat grown in the Netherlands dates from 2001. This aim of this study was to investigate the incidence and levels of Fusarium Head Blight species and Fusarium mycotoxins, as well as their possible relationships, in winter wheat cultivated in the Netherlands in 2009. Samples were collected from individual fields of 88 commercial wheat growers. Samples were collected at harvest from 86 fields, and 2 weeks before the expected harvest date from 21 fields. In all, 128 samples, the levels of each of seven Fusarium Head Blight species and of 12 related mycotoxins were quantified. The results showed that F. graminearum was the most frequently observed species at harvest, followed by F. avenaceum and M. nivale. In the pre-harvest samples, only F. graminearum and M. nivale were relevant. The highest incidence and concentrations of mycotoxins were found for deoxynivalenol, followed by zearalenone and beauvericin, both pre-harvest and at harvest. Other toxins frequently found--for the first time in the Netherlands--included T-2 toxin, HT-2 toxin, and moniliformin. The levels of deoxynivalenol were positively related to F. graminearum levels, as well as to zearalenone levels. Other relationships could not be established. The current approach taken in collecting wheat samples and quantifying the presence of Fusarium Head Blight species and related mycotoxins is an efficient method to obtain insight into the occurrence of these species and toxins in wheat grown under natural environmental conditions. It is recommended that this survey be repeated for several years to establish inter-annual variability in both species composition and mycotoxin occurrence.

Topics: Chromatography, High Pressure Liquid; Crops, Agricultural; Cyclobutanes; Depsipeptides; Food Contamination; Fusarium; Limit of Detection; Mycotoxins; Netherlands; Plant Diseases; Reproducibility of Results; Seeds; Species Specificity; Spectrometry, Mass, Electrospray Ionization; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Triticum; Zearalenone

Mycotoxins are secondary metabolites produced by filamentous fungi that cause a toxic response when ingested by animals or man. Demand of natural fur, such as those from rabbit and chinchilla, produced under controlled conditions, has increased worldwide. The toxicogenic mycoflora contaminating feeds for these animals was enumerated and identified. Six of the major mycotoxins implicated in animal mycotoxicosis were detected and quantified. Moulds count ranged from <10 to 4.7 × 10(5) CFU g(-1); 14% of the samples exceeded the limit that determines hygienic feed quality. More than twenty species belonging to the five most important mycotoxigenic mould genera were recovered. Among the analyzed mycotoxins, aflatoxins were recovered in 100% of the examined samples, deoxynivalenol in 95%, fumonisins in 100%, ochratoxin A in 98%, T2 toxin in 98%, and zearalenone in 100%. Cooccurrence of mycotoxins was observed in 100% of the samples analyzed. Exposure to multiple mycotoxins was thus demonstrated for these animals.

Topics: Aflatoxins; Animal Feed; Animals; Chinchilla; Food Contamination; Fumonisins; Fungi; Mycotoxicosis; Mycotoxins; Ochratoxins; Rabbits; T-2 Toxin; Trichothecenes; Zearalenone

The predominant species in maize in temperate climates is Fusarium graminearum, which produces the mycotoxins deoxynivalenol and zearalenone. Projected climate change is expected to affect Fusarium incidence and thus the occurrence of these mycotoxins. Predictive models may be helpful in determining trends in the levels of these mycotoxins with expected changing climatic conditions. The aim of this study was to develop a model describing fungal infection and subsequent growth as well as the formation of deoxynivalenol and zearalenone in maize in The Netherlands. For this purpose, a published Italian model was used as a starting point. This model is a mixed empiric-mechanistic model that describes fungal infection during silking (based on wind speed and rainfall) and subsequent germination, growth and toxin formation (depending on temperature and water availability). Model input uses weather parameters and crop management factors, such as maize hybrid, sowing date, flowering period and harvest date. Model parameter values were obtained by fitting these parameters to deoxynivalenol and zearalenone measurements in Dutch maize, using national mycotoxin data from the years 2002-2007. The results showed that the adapted model is capable of describing the trend in average deoxynivalenol and zearalenone levels over these years. Validation with external data is needed to verify model outcomes. It is expected that the current model can be used to estimate the effect of projected climate change on trends in deoxynivalenol and zearalenone levels in the coming years.

Topics: Agriculture; Animals; Climate Change; Crops, Agricultural; Flowering Tops; Food Contamination; Forecasting; Fusarium; Germination; Humans; Models, Biological; Mycotoxins; Netherlands; Seasons; Seeds; Species Specificity; Trichothecenes; Weather; Zea mays; Zearalenone

Mycotoxins, together with endotoxins, represent important classes of naturally occurring contaminants in food products, posing significant health risks to consumers. The aim of this study is to investigate the occurrence of both Fusarium mycotoxins and endotoxins in commercially produced traditional banana beer. Two brands of commercially produced traditional banana beer were collected from a local retail market in Kigali, Rwanda. Beer samples were analysed for the presence of deoxynivalenol (DON), fumonisin B₁ and zearalenone (ZEA), using an enzyme-linked immuno-sorbent assay (ELISA) method. The quantification of bacterial endotoxin using Limulus amoeboecyte lysate (LAL) assay was also conducted. The contamination levels were 20 and 6.7 µg kg⁻¹ for DON; 34 and 31.3 µg kg⁻¹ for FB₁; 0.66 and 2.2 µg kg⁻¹ for ZEA in brands A and B of the beers, respectively. Results indicate that the levels of Fusarium toxins and bacterial endotoxin reported in this study did not pose adverse human health effects as a result of drinking/consuming banana beer. However, exposure to low/sub-threshold doses or non-toxic levels of endotoxins magnifies the toxic effect of xenobiotic agents (e.g. fungal toxins) on liver and other target organs. Considering Fusarium toxins and/or endotoxin contamination levels in other agricultural commodities intended for human consumption, health risks might be high and the condition is aggravated when beer is contaminated by mixtures of the mycotoxins, as indicated in this study.

Topics: Beer; Diet; Endotoxins; Enzyme-Linked Immunosorbent Assay; Food Contamination; Food Inspection; Fruit; Fumonisins; Fusarium; Humans; Limulus Test; Musa; Mycotoxins; Rwanda; Trichothecenes; Zearalenone

This study aimed to investigate mycotoxin contamination of cereal grain commodities for feed and food production in North Western Europe during the last two decades, including trends over time and co-occurrence between toxins, and to assess possible effects of climate on the presence of mycotoxins. For these aims, analytical results related to mycotoxin contamination of cereal grain commodities, collected in the course of national monitoring programmes in Finland, Sweden, Norway and the Netherlands during a 20-year period, were gathered. Historical observational weather data, including daily relative humidity, rainfall and temperature, were obtained from each of these four countries. In total 6382 records, referring to individual sample results for mycotoxin concentrations (one or more toxins) in cereal grains were available. Most records referred to wheat, barley, maize and oats. The most frequently analysed mycotoxins were deoxynivalenol, 3-acetyl-deoxynivalenol, nivalenol, T-2 toxin, HT-2 toxin and zearalenone. Deoxynivalenol had the highest overall incidence of 46%, and was mainly found in wheat, maize and oats. Mycotoxins that showed co-occurrence were: deoxynivalenol and 3-acetyl-deoxynivalenol in oats; deoxynivalenol and zearalenone in maize and wheat; and T-2 toxin and HT-2 toxin in oats. The presence of both deoxynivalenol and zearalenone in wheat increased with higher temperatures, relative humidity and rainfall during cultivation, but the presence of nivalenol was negatively associated with most of these climatic factors. The same holds for both nivalenol and deoxynivalenol in oats. This implies that climatic conditions that are conducive for one toxin may have a decreasing effect on the other. The presence of HT-2 toxin in oats showed a slight decreasing trends over time, but significant trends for other toxins showed an increasing presence during the last two decades. It is therefore useful to continue monitoring of mycotoxins. Obtained results can be used for development of predictive models for presence of mycotoxins in cereal grains.

Topics: Acetylation; Agriculture; Animals; Climate Change; Crops, Agricultural; Edible Grain; Food Contamination; Fusarium; Humans; Mycotoxins; Netherlands; Scandinavian and Nordic Countries; Seeds; Spatio-Temporal Analysis; T-2 Toxin; Trichothecenes; Weather; Zearalenone

Exposure to multiple mycotoxins through the food chain represents a major potential health hazard to both humans and livestock. They can cause a variety of severe acute as well as chronic diseases. Eliminating mycotoxins from various grain crops is a global health priority. According to the Food and Agriculture Organization (FAO), world food production needs to double by 2050. Innovative solutions will be required to sustain toxin free grain supplies worldwide.. A competitive flow cytometry based multiplexed assay with fluorescent microspheres has been developed. The new multiplexed method can analyze simultaneously any one or all six major mycotoxins. They include: Ochratoxin A (OTA), Aflatoxin B1 (AFB1), Fumonisin B1 (FB1), T-2 toxin (T-2), Deoxynivalenol (DON) and Zearalenone (ZEA), which are all potential human health hazards. The CFIA described here includes a simplified single extraction step for mycotoxins from specimens and a comprehensive post acquisition software module. The new assay system was developed with a FACSArray™ BD Bioanalyzer flow cytometer (BD Biosciences, Belgium).. The CFIA performs favourably when compared to commercial ELISA. Sensitivity range with CFIA increased between 13% and 100% with an average improvement of 50% for the six mycotoxins.. The multiplexed assay presented here has the unique capacity to quantify up to six mycotoxins simultaneously from a single specimen extraction. CFIA's poly-mycotoxin detection sensitivity exceeds standard ELISA. CFIA may be part of a comprehensive assay system that will provide reliable and effective safeguard for agricultural commodities to be free of mycotoxins.

Topics: Aflatoxin B1; Animals; Antibodies, Fungal; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Fluorescent Dyes; Fumonisins; Humans; Immunoassay; Mice; Microspheres; Mycotoxins; Ochratoxins; Reproducibility of Results; Sensitivity and Specificity; T-2 Toxin; Trichothecenes; Zearalenone

Mycotoxin contaminations pose a growing problem in animal production from the economic and toxicological point of view. Clinical symptoms of mycotoxicosis are relatively unspecific, making the disease difficult to diagnose. This study presents a clinical case of dairy cattle infected with natural mycotoxins produced by fungi of the genus Fusarium (zearalenone [ZEA] and deoxynivalenol [DON]) in eastern Poland. In dead and infected cows, the presence of ZEA and DON was determined in the blood serum, significant changes were observed in blood morphological and biochemical profiles, extravasations and bowel inflammations were also observed. The results reported testify to an acute autoimmune process in the intestines as well as immunosuppression.

Topics: Animals; Cattle; Cattle Diseases; Dairying; Female; Mycotoxicosis; Poland; Trichothecenes; Zearalenone

Mycotoxins such as beauvericin (BEA), deoxynivalenol (DON), enniatin B (ENB), fumonisin B1 (FB1), T-2 toxin and zearalenone (ZEA) can co-occur in food commodities. This aim of this study was to assess the myelotoxicity of these mycotoxins in couple using in vitro human granulo-monocytic (Colony Forming Unit-Granulocyte and Macrophage, CFU-GM) hematopoietic progenitors. Clonogenic assays have been performed in the presence of the following couples of fusariotoxins: DON + BEA, DON + FB1, DON + T-2, DON + ZEA, T-2 + ZEA and BEA + ENB. Co-exposure of human CFU-GM to DON + BEA resulted in synergic myelotoxic effects. The combination of DON + T-2 presented additive or synergic myelotoxic effects. The couples DON + ZEA, T-2 + ZEA and BEA + ENB had additive myelotoxic effects, while the combination of DON + FB1 showed antagonist myelotoxic effects. These in vitro results suggested that the simultaneous presence of mycotoxins in food commodities and diet may be more myelotoxic than the presence of one mycotoxin alone. Diminution of hematopoietic progenitors could give rise to a decrease number of mature blood cells, inducing agranulocytosis and/or thrombocytopenia and in severe cases aplastic anemia.

Topics: Cells, Cultured; Depsipeptides; Fumonisins; Fusarium; Granulocyte-Macrophage Progenitor Cells; Humans; Mycotoxins; T-2 Toxin; Toxicity Tests; Trichothecenes; Zearalenone

A total of 92 commercial compound feeds from South Africa were investigated for various mycotoxins. The data reveal the highest incidence of feed contamination for fumonisins (FB) (range: 104-2999 µg/kg) followed by deoxynivalenol (DON) (range: 124-2352 µg/kg) and zearalenone (ZEA) (range: 30-610 µg/kg). The incidence of ochratoxin A (OTA) and aflatoxins (AF)-contaminated samples were generally low, i.e., 4% and 30% of samples with levels ranging between 6.4 and 17.1 µg/kg (mean: 9.9 µg/kg) for OTA and 0.2 to 71.8 µg/kg (mean: 9.0 µg/kg) for AF. No samples contained T-2 toxin or HT-2 toxin. However, all samples analyzed were contaminated with at least one mycotoxin with a majority containing several mycotoxins. In particular, 3 of 4 positive samples mainly cattle feeds that had relatively high contents of OTA (ranging from 7 to 17.1 µg/kg) also contained high amounts of AF (>27.5 µg/kg) together with FB, DON and ZEA. Apart from a few samples, the levels of mycotoxins may be regarded as safe for livestock production in South Africa. However, the persistent co-occurrence of mycotoxins in samples, especially those at high concentrations, i.e., AF and OTA, together with other mycotoxins studied, may elicit synergistic or additive effects in animals. This should raise concern as multiple contaminations may pose a risk to livestock production and health.

Topics: Aflatoxins; Animal Feed; Animals; Cattle; Chromatography, Liquid; Food Contamination; Food Microbiology; Fumonisins; Mycotoxins; Ochratoxins; South Africa; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

Fusarium spp. invasion causes head blight, a destructive disease in the world's main wheat-growing areas, and deoxynivalenol (DON) and zearalenone (ZEA) contamination in cereal-based products. No data are available on the relationship between Fusarium spp. on commercial wheat samples in Mexico City and the presence of mycotoxins. A total of 30 wheat samples were subject to a PCR method involving genes of the trichothecene and zearalenone biosynthesis pathways to detect the presence of Fusarium. Detection and quantification of DON and ZEA was performed using liquid chromatography coupled to UV detection. PCR indicated the presence of the Tri5 and PKS4 genes in 16.7 and 23.3% of samples, respectively. DON and ZEA contamination was found in 51.2 and 71.4% of samples, respectively, where a positive amplification was obtained. This work presents up-to-date information on mycotoxin contamination in Mexico, where improved contamination/exposure data and firm control/monitoring measures are needed.

Topics: Carbon-Carbon Lyases; Diet; Food Contamination; Food Inspection; Fungal Proteins; Fusarium; Guideline Adherence; Health Policy; Health Promotion; Humans; Mexico; Molecular Typing; Mycological Typing Techniques; Poisons; Risk Assessment; Seeds; Trichothecenes; Triticum; Zearalenone

The health risks of deoxynivalenol (DON) and zearalenone (ZEN) necessitate the development of analytical methods for widespread food and feed screening. We sought to establish a rapid, economic and sensitive dual-label time-resolved fluoroimmunoassay (TRFIA) to detect DON and ZEN simultaneously. Eu(3+)- and Sm(3+)-labelled antibodies were used, as lanthanides are more stable and have narrower emission spectra than most fluorescent dyes.. The limit of detection was 0.0194 ng mL(-1) for DON (range: 0.0194-100 ng mL(-1)) and 0.37 ng mL(-1) for ZEN (range: 0.37-50 ng mL(-1)). DON recovery in spiked cereal samples was 88-107%, and for ZEN was 83-108%, with both intra- and inter-assay coefficients of variation (CVs) less than 5%. The dual-label TRFIA results correlated well with ELISA results (correlation coefficients: 0.9733 for DON and 0.9784 for ZEN).. The dual-label TRFIA is a simple, fast and sensitive method for high-throughput screening of DON and ZEN in food and feedstuff.

Topics: Antibodies; Edible Grain; Enzyme-Linked Immunosorbent Assay; Fluoroimmunoassay; Food Contamination; Food Safety; Fungi; Limit of Detection; Trichothecenes; Zearalenone

The issue of moulds and, thus, contamination with mycotoxins is very topical, particularly in connexion with forages from grass stands used at the end of the growing season. Deoxynivalenol (DON), zearalenone (ZEA), fumonisins (FUM) and aflatoxins (AFL) are among the most common mycotoxins. The aim of the paper was to determine concentrations of mycotoxins in selected grasses (Lolium perenne, Festulolium pabulare, Festulolium braunii) and their mixtures with Festuca rubra an/or Poa pratensis during the growing season as a marker of grass safety, which was assessed according to content of the aforementioned mycotoxins. During the growing season grass forage was contaminated with mycotoxins, most of all by DON and ZEA. The contents of AFL and FUM were zero or below the limit of quantification. Moreover, the level of the occurrence of mould was quantified as ergosterol content, which was higher at the specific date of cut. All results were statistically processed and significant changes were discussed.

Topics: Aflatoxins; Animal Feed; Czech Republic; Enzyme-Linked Immunosorbent Assay; Ergosterol; Festuca; Food Contamination; Food Microbiology; Fumonisins; Fungi; Lolium; Mycotoxins; Poa; Poaceae; Poisons; Seasons; Trichothecenes; Zearalenone

A prototype imaging surface plasmon resonance-based multiplex microimmunoassay for mycotoxins is described. A microarray of mycotoxin-protein conjugates was fabricated using a continuous flow microspotter device. A competitive inhibition immunoassay format was developed for the simultaneous detection of deoxynivalenol (DON) and zearalenone (ZEN), using a single sensor chip. Initial in-house validation showed limits of detection of 21 and 17 ng/mL for DON and 16 and 10 ng/mL for ZEN in extracts, which corresponds to 84 and 68 μg/kg for DON and 64 and 40 μg/kg for ZEN in maize and wheat samples, respectively. Finally, the results were critically compared with data obtained from liquid chromatography-mass spectrometry confirmatory analysis method and found to be in good agreement. The described multiplex immunoassay for the rapid screening of several mycotoxins meets European Union regulatory limits and represents a robust platform for mycotoxin analysis in food and feed samples.

Topics: Immunoassay; Limit of Detection; Mycotoxins; Surface Plasmon Resonance; Trichothecenes; Triticum; Zea mays; Zearalenone

We developed a method for the simultaneous determination of deoxynivalenol, T-2 toxin, HT-2 toxin and zearalenone in wheat and biscuit by liquid chromatography/electrospray ionization/tandem mass spectrometry coupled with immunoaffinity extraction. This chapter describes a method to extract, clean-up, and quantitate these mycotoxins and the effect of the ion suppression of multifunctional column and IAC in the clean-up were compared.

Topics: Chromatography, Liquid; Food Analysis; Food Contamination; Mycotoxins; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

It was hypothesized that a mycotoxin binder, Grainsure E, would inhibit adverse effects of a mixture of fumonisin B1, deoxynivalenol, and zearalenone in rats. For 14 and 28 days, 8-10 Sprague-Dawley rats were fed control diet, Grainsure E (0.5%), toxins (7 μg fumonisin B1/g, 8 μg of deoxynivalenol/g and 0.2 μg of zearalenone/g), toxins (12 μg of fumonisin B1/g, 9 μg of deoxynivalenol/g, and 0.2 μg of zearalenone/g + Grainsure E), or pair-fed to control for food intake of toxin-fed rats. After 28 days, decreased body weight gain was prevented by Grainsure E in toxin-fed female rats, indicating partial protection against deoxynivalenol and fumonisin B1. Two effects of fumonisin B1 were partly prevented by Grainsure E in toxin-fed rats, increased plasma alanine transaminase (ALT) and urinary sphinganine/sphingosine, but sphinganine/sphingosine increase was not prevented in females at the latter time point. Grainsure E prevented some effects of fumonisin B1 and deoxynivalenol in rats.

Topics: Animals; Chemical and Drug Induced Liver Injury; Diet; Female; Fumonisins; Kidney Diseases; Liver Diseases; Male; Mycotoxins; Rats; Rats, Sprague-Dawley; Trichothecenes; Zearalenone

Rapid and sensitive methods to detect Fusarium culmorum and trichothecene and zearalenone producing strains in food and feed are valuable in predicting potential contamination. In this study the effectiveness of primers, recommended in the literature, for species identification of F. culmorum and basic genes encoding for mycotoxin production was tested. A total of 68 isolates of F. culmorum were collected from cereals and potato between 2005 and 2008 from different Polish provinces. It was shown that from among the four primer pairs enabling the identification of F. culmorum, and therefore also to establish its presence in the material, only primers Fc01F/Fc01R seem to be fully effective in the case of Polish strains. Determination of material contamination by F. culmorum, however, is only a first step in determining food safety. It is also extremely important to identify genes encoding the potential ability to produce mycotoxins. It was shown that three pairs of primers (tox5-1/tox5-2, HATriF/HATriR and Tri5F/Tri5R) enable a fully effective identification of the presence of the Tri5 gene responsible for producing trichothecenes. Determination of the DON-chemotype, and thus identification of the strains of F. culmorum potentially producing deoxynivalenol, is enabled equally by MinusTri7F/MinusTri7F, Tri7F/Tri7DON and Tri13F/Tri13DONR. However, a determination of the NIV-chemotype, and thus identification of the strains potentially producing nivalenol, is enabled by Tri7F/Tri7R, Tri7F/Tri7NIV and Tri13NIVF/Tri13R. The potential ability of isolates to produce ZEA can be determined to the same degree in assay with PKS4-PS.1/PKS4-PS.2 and F1/R1.

Topics: Base Sequence; DNA Primers; DNA, Fungal; Food Contamination; Food Microbiology; Fusarium; Genes, Fungal; Mycotoxins; Polymerase Chain Reaction; Species Specificity; Trichothecenes; Zearalenone

To quantify and to compare the occurrence of Fusarium species in maize kernels and stalk pieces, to analyse mycotoxins in kernels and maize crop residues, to evaluate two approaches to obtain kernel samples and to compare two methods for mycotoxin analyses.. The occurrence of Fusarium species in maize kernels and stalk pieces from a three-year maize hybrid trial and 12 kernel samples from grower's fields was assessed. Nine to 16 different Fusarium species were detected in maize kernels and stalks. In kernels, F. graminearum, F. verticillioides and F. proliferatum were the most prevalent species whereas in stalks, they were F. equiseti, F. proliferatum and F. verticillioides. In 2006, 68% of the kernel samples exceeded the recommended limit for pig feed for deoxynivalenol (DON) and 42% for zearalenone (ZON), respectively. Similarly, 75% of the samples from grower's fields exceeded the limits for DON and 50% for ZON. In maize crop residues, toxin concentrations ranged from 2.6 to 15.3 mg kg(-1) for DON and from 0.7 to 7.4 mg kg(-1) for ZON. Both approaches to obtain maize kernel samples were valid, and a strong correlation between mycotoxin analysis using ELISA and LC-MS/MS was found.. The contamination of maize kernels, stalk pieces and remaining crop residues with various mycotoxins could pose a risk not only to animal health but also to the environment. With the hand-picked sample, the entire Fusarium complex can be estimated, whereas combine harvested samples are more representative for the mycotoxin contents in harvested goods.. This is the first multi-year study investigating mycotoxin contamination in maize kernels as well as in crop residues. The results indicate a high need to identify cropping factors influencing the infection of maize by Fusarium species to establish recommendations for growers.

Topics: Chromatography, Liquid; Enzyme-Linked Immunosorbent Assay; Food Contamination; Fusarium; Plant Stems; Seeds; Switzerland; Tandem Mass Spectrometry; Trichothecenes; Zea mays; Zearalenone

A total of 201 samples of brown rice, polished rice, and two types of by-products, blue-tinged rice and discolored rice, were collected from rice stores maintained at 51 rice processing complexes in Korea. These samples were analyzed for the presence of Fusarium mycotoxins such as deoxynivalenol (DON), nivalenol (NIV), and zearalenone (ZEA). Contaminants (and their ranges) found in discolored rice samples were DON (59 to 1,355 ng g(-1)), NIV (66 to 4,180 ng g(-1)), and ZEA (25 to 3,305 ng g(-1)); those found in blue-tinged (less-ripe) rice were DON (86 to 630 ng g(-1)), NIV (50 to 3,607 ng g(-1)), and ZEA (26 to 3,156 ng g(-1)). Brown rice samples were contaminated mostly with NIV and ZEA (52 to 569 ng g(-1) and 47 to 235 ng g(-1), respectively). Polished rice samples were largely free from mycotoxins, although one sample was contaminated with NIV (77 ng g(-1)). When the fungal flora associated with each rice sample was investigated, blue-tinged rice was the most often contaminated with Fusarium graminearum (3.8%), followed by the discolored rice (2.4%) and brown rice (1.6%) samples. Using PCR, toxin genotyping of 266 isolates of F. graminearum revealed that most isolates (96%) were NIV producers. In conclusion, this survey is the first report of the cocontamination of Korean rice and its by-products with trichothecenes and ZEA. Importantly, it also provides new information on the natural contamination of rice by Fusarium mycotoxins.

Topics: Consumer Product Safety; Food Contamination; Food Microbiology; Fusarium; Humans; Korea; Mycotoxins; Oryza; Prevalence; Trichothecenes; Zearalenone

Seven commercially available deoxynivalenol (DON) and zearalenone (ZEN) immunoaffinity columns (IACs) were tested for cross-reactivity to conjugated forms (3-acetyl-deoxynivalenol, 15-acetyl-deoxynivalenol, DON-3-glucoside, DON-3-glucuronide, ZEN-glucosides, ZEN-glucuronide) and metabolites (de-epoxydeoxynivalenol, α-zearalenol, β-zearalenol) and nivalenol (NIV), using a semi-quantitative multi-mycotoxin ultra-performance liquid chromatography-tandem mass spectrometry method. The DON IACs showed cross-reactivity for nearly all DON derivatives tested. The ZEN IACs showed limited cross-reactivity to some of the ZEN derivatives. The IACs were evaluated for their potential use as sample clean-up for mycotoxins in serum.

Topics: Animals; Antibody Specificity; Cattle; Chromatography, Affinity; Chromatography, High Pressure Liquid; Immunoassay; Serum; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

Fusarium graminearum is the most important pathogen causing Fusarium head blight (FHB) of small cereal grains worldwide responsible for quantitative and qualitative yield losses. The presence in crops is often associated with mycotoxin contamination of foodstuff limiting its use for human and animal consumption. A collection of isolates of F. graminearum from Germany was characterized genetically and chemically for their potential to produce the B trichothecenes deoxynivalenol (DON) and nivalenol (NIV). Molecular methods with eight PCR assays were implemented based on functional Tri7 and Tri13 genes and on the tri5-tri6 intergenic region to differentiate between chemotaxonomic groups DON and NIV, resulting in a marked majority (61/63) of DON chemotypes. Mycotoxins produced on rice kernels were quantified by means of LC-MSMS including DON, NIV, 3-acetyl-DON (3-ADON), 15-acetyl-DON (15-ADON), DON-3-glucoside, fusarenon X, as well as zearalenone; all of them proving to be present in high concentration among the isolates. All DON-chemotype isolates also produced lower amounts of NIV with the amount being positively correlated (R²=0.89) to the DON amount. 15-ADON and 3-ADON are reported to be produced simultaneously by the isolates, the former dominating over the latter in all but one isolate. Fungal biomass, was quantified via ergosterol amount on rice. It was used to calculate specific mycotoxin production per biomass of isolates, ranging from 0.104 to 1.815mg DON mg-1 ergosterol, presenting a Gaussian distribution. Genotype and phenotype characterization revealed discrepancies with respect to mycotoxin production potential of the fungi, i.e. isolates from one chemotype were able to produce mycotoxins from other chemotypes in considerable amounts.

Topics: DNA, Fungal; Ergosterol; Fusarium; Genotype; Germany; Glucosides; Oryza; Phenotype; Polymerase Chain Reaction; Trichothecenes; Zearalenone

The EU has set maximum limits for the Fusarium mycotoxins, deoxynivalenol (DON) and zearalenone (ZON). The maximum permitted level decreases from unprocessed wheat, through intermediary products, e.g. flour, to finished products such as bakery goods and breakfast cereals. It is, therefore, important to understand the effects of processing on the mycotoxin distribution in mill fractions. Between 2004 and 2007, samples were taken at commercial flour mills at various points in the milling process and analysed for trichothecenes and ZON. Samples with a range of mycotoxin concentrations harvested in 2004 and 2005 were processed in a pilot mill and the mycotoxins in the different mill fractions quantified. In the commercial samples, DON was the predominant mycotoxin with highest levels detected in the bran fraction. Analysis of the pilot mill fractions identified a significant difference between the two years and between mycotoxins. The proportion of DON and nivalenol in the mill fractions varied between years. DON and nivalenol were higher in flour fractions and lower in bran and offal in samples from 2004 compared to samples from 2005. This may be a consequence of high rainfall pre-harvest in 2004 resulting in movement of these mycotoxins within grains before harvest. There was no significant difference in the distribution of ZON within mill fractions between the two years. For DON, higher concentrations in the grain resulted in a greater proportion of DON within the flour fractions. Understanding the factors that impact on the fractionation of mycotoxins during milling will help cereal processors to manufacture products within legislative limits.

Topics: Flour; Food Contamination; Food Handling; Fusarium; Mycotoxins; Seeds; Trichothecenes; Triticum; United Kingdom; Zearalenone

Maize is frequently infected by the Fusarium species producing mycotoxins. Numerous investigations have focused on grain maize, but little is known about the Fusarium species in the entire plant used for silage. Furthermore, mycotoxins persist during the ensiling process and thus endanger feed safety. In the current study, we analyzed 20 Swiss silage maize samples from growers' fields for the incidence of Fusarium species and mycotoxins. The species spectrum was analyzed morphologically and mycotoxins were measured by LC-MS/MS. A pre-harvest visual disease rating showed few disease symptoms. In contrast, the infection rate of two-thirds of the harvest samples ranged from 25 to 75% and twelve different Fusarium species were isolated. The prevailing species were F. sporotrichioides, F. verticillioides and F. graminearum. No infection specificity for certain plant parts was observed. The trichothecene deoxynivalenol (DON) was found in each sample (ranging from 780 to 2990 µg kg(-1)). Other toxins detected in descending order were zearalenone, further trichothecenes (nivalenol, HT-2 and T-2 toxin, acetylated DON) and fumonisins. A generalized linear regression model containing the three cropping factors harvest date, pre-precrop and seed treatment was established, to explain DON contamination of silage maize. Based on these findings, we suggest a European-wide survey on silage maize.

Topics: Chromatography, Liquid; Food Contamination; Fumonisins; Fusarium; Mycotoxins; Silage; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zea mays; Zearalenone

The aim of the present experiment was to investigate the effects of feeding grains naturally contaminated with Fusarium mycotoxins on morphometric indices of jejunum and to follow the passage of deoxynivalenol (DON) through subsequent segments of the digestive tract of broilers. A total of 45 1-d-old broiler chickens (Ross 308 males) were randomly allotted to three dietary treatments (15 birds/treatment): (1) control diet; (2) diet contaminated with 1 mg DON/kg feed; (3) diet contaminated with 5 mg DON/kg feed for five weeks. None of the zootechnical traits (body weight, body weight gain, feed intake, and feed conversion) responded to increased DON levels in the diet. However, DON at both dietary levels (1 mg and 5 mg DON/kg feed) significantly altered the small intestinal morphology. In the jejunum, the villi were significantly (P < 0.01) shorter in both DON treated groups compared with the controls. Furthermore, the dietary inclusion of DON decreased (P < 0.05) the villus surface area in both DON treated groups. The absolute or relative organ weights (liver, heart, proventriculus, gizzard, small intestine, spleen, pancreas, colon, cecum, bursa of Fabricius and thymus) were not altered (P > 0.05) in broilers fed the diet containing DON compared with controls. DON and de-epoxy-DON (DOM-1) were analyzed in serum, bile, liver, feces and digesta from consecutive segments of the digestive tract (gizzard, cecum, and rectum). Concentrations of DON and its metabolite DOM-1 in serum, bile, and liver were lower than the detection limits of the applied liquid chromatography coupled with mass spectrometry (LC-MS/MS) method. Only about 10 to 12% and 6% of the ingested DON was recovered in gizzard and feces, irrespective of the dietary DON-concentration. However, the DON recovery in the cecum as percentage of DON-intake varied between 18 to 22% and was not influenced by dietary DON-concentration. Interestingly, in the present trial, DOM-1 did not appear in the large intestine and in feces. The results indicate that deepoxydation in the present study hardly occurred in the distal segments of the digestive tract, assuming that the complete de-epoxydation occurs in the proximal small intestine where the majority of the parent toxin is absorbed. In conclusion, diets with DON contamination below levels that induce a negative impact on performance could alter small intestinal morphology in broilers. Additionally, the results confirm that the majority of the ingested DON qui

Topics: Animal Feed; Animals; Chickens; Chromatography, Liquid; Food Contamination; Food Microbiology; Fusarium; Intestine, Small; Male; Organ Size; Tandem Mass Spectrometry; Tissue Distribution; Trichothecenes; Zearalenone

The aim of the research was to describe the influence of selected mycotoxins on major factors (alcohol concentration, productivity, yield and energy) that are characteristic of the fermentation process of maize mashes. Indicators of the alcoholic fermentation of mashes made from raw material with low contaminations levels were compared with mashes obtained from raw material that was selectively contaminated with mycotoxins on the following concentrations: aflatoxin B(1)-11.65 ppb, B(2)-12.60 ppb, G(1)-12.34 ppb, G(2)-12.04 ppb; ochratoxin A-177.5 ppb; zearalenone-352 ppb; deoxynivalenol-2274 ppb; fumonisin B(1)-1875 ppb, B(2)-609 ppb, B(3)-195 ppb. It was found that, apart from fumonisin, all mycotoxins substantially affected the course of subsequent fermentation phases, in particular the first and the main fermentation phases. The highest drop in alcohol concentration at the main stage of the process amounted to 1% v/v and it was achieved by contamination with zearalenone. The statistically significant drop in the final fermentation yield was observed; this was caused by raw material contaminated with all studied mycotoxins, except for fumonisin. The decrease in ethanol yield in reference to the control variant ranged from 1.42 to 3.20 dm(3) of absolute alcohol out of 100 kg of starch, depending on a toxin.

Topics: Aflatoxins; Alcohols; Fermentation; Food Contamination; Fumonisins; Mycotoxins; Ochratoxins; Trichothecenes; Zearalenone

A new analytical method for the rapid and simultaneous determination of five mycotoxins (zearelenone, deoxynivalenol, Fusarenon X, 15-acetyldeoxynivalenol and nivalenol) in breakfast cereals and flours by heart-cutting GC-MS has been developed and validated. Extraction was performed with MeCN, applying a modified QuEChERS (QUick, Easy, CHeap, Effective, Rugged and Safe) procedure, and the extracts were analyzed after a silylation of the analytes under study. Careful optimization of the parameters of Deans Switch device and GC-MS was achieved in order to attain a fast separation in SIM mode, allowing a total run time of only 8 min. Acceptable recoveries for all mycotoxins at two different spiking levels (20 and 100 microg/kg) were achieved with good repeatability (from 9 to 21%). LOD ranged from 2 to 15 microg/kg and LOQ ranged from 5 to 50 microg/kg, which were lower than the maximum limit legal established by the European Union (EU). The method developed was applied to commercial breakfast cereals and flours; among the mycotoxins studied, deoxynivalenol and zearalenone were the most predominant.

Topics: Edible Grain; Flour; Gas Chromatography-Mass Spectrometry; Molecular Structure; Mycotoxins; Reproducibility of Results; Safety; Time Factors; Trichothecenes; Zearalenone

A collection of 84 cereal-based food products in 25 composites, including beer, was screened for the presence of deoxynivalenol, zearalenone, and their respective metabolites deoxynivalenol-3-glucopyranoside, 3-acetyl-deoxynivalenol, zearalenol-4-glucopyranoside, alpha-zearalenol, beta-zearalenol, alpha-zearalenol-4-glucopyranoside, beta-zearalenol-4-glucopyranoside, and zearalenone-4-sulfate. The most abundant analyte was zearalenone-4-sulfate, which was found in 13 composites, albeit in low concentrations. Furthermore, deoxynivalenol was detected in eight, zearalenone in seven, and deoxynivalenol-3-glucopyranoside in two composites. None of the remaining six analytes was found in any matrices, which suggests that, if at all present, the concentrations of these latter metabolites are very low and, hence, do not impose any danger to consumers. The highest mycotoxin content was found in bran flakes with 254 ng g(-1) deoxynivalenol, 6 ng g(-1) zearalenone-4-sulfate, and 44 ng g(-1) zearalenone.

Topics: Austria; Chromatography, High Pressure Liquid; Edible Grain; Fast Foods; Food Contamination; Food Inspection; Fusarium; Limit of Detection; Mycotoxins; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Trichothecenes; United Kingdom; Zearalenone

The effects of aflatoxin B₁ (AFB₁), aflatoxin M₁ (AFM₁), deoxynivalenol (DON) and zearalenone (ZEA) on the viability, chemiluminescent (CL) response and expression of cytokine mRNA of bovine neutrophils (PMNs) were evaluated. The opsonized zymosan (OPZ)-stimulated CL response of PMNs was significantly (P < 0.05) decreased by AFB₁ ( > 50 pg/ml), AFM₁ ( > 50 pg/ml) and ZEA (>50 pg/ml). The phorbol myristate acetate (PMA)-stimulated CL response PMNs was significantly (P < 0.05) decreased by AFB₁ (> 0.5 pg/ml), AFM₁ (> 50 pg/ml), ZEA (> 500 pg/ml) and DON ( > 5 pg/ml). Treatment with AFB₁ resulted in reduction in the mRNA expression of interleukin-1β and tumor necrosis factor-α of PMNs stimulated with OPZ and PMA. These results suggest that these four mycotoxins have inhibitory effects on the function of bovine PMNs.

Topics: Aflatoxin B1; Aflatoxin M1; Animals; Cattle; Cell Survival; Cytokines; Gene Expression Regulation; Kinetics; Luminescence; Mycotoxins; Neutrophils; RNA, Messenger; Trichothecenes; Zearalenone

Two groups of pregnant primiparous sows (day 89 ± 2 of gestation) were 54 days (± 1 day) fed either with an experimental diet (5.08 mg kg(-1) deoxynivalenol--DON, 0.09 mg kg(-1) zearalenone and 21.61 mg kg(-1) fusaric acid) or control diet (0.25 mg kg(-1) DON). The consummation of feed was significantly higher in the control group. Lymphocyte stimulation in culture from peripheral blood lymphocytes (PBL) was measured by BrdU incorporation test using two different concentrations of mitogen PHA 10 and 20 μg ml(-1), two different concentrations of DON (5 and 1 μg ml(-1)) and a combination of both, PHA and DON (PHA 10+DON 5, PHA 10+DON 1 and PHA 10+DON 0.1 μg ml(-1)). The lymphocyte proliferation, except for PHA 10 μg ml(-1), was significantly higher in the experimental group. Further on, using the photometric enzyme immunoassay, the apoptosis was studied in non-stimulated 72h lymphocyte culture or stimulated with 1 μg ml(-1) of DON. The significantly higher apoptosis was in non-stimulated lymphocyte cultures from the experimental group (P = 0.036). The results suggest that such feed may affect the peripheral lymphocyte population in the direction of their proliferation response and programmed cell death.

Topics: Algorithms; Animal Feed; Animals; Animals, Newborn; Antimetabolites; Apoptosis; Body Weight; Bromodeoxyuridine; Cell Proliferation; Cells, Cultured; Diet; Eating; Enzyme-Linked Immunosorbent Assay; Female; Food Contamination; Fusaric Acid; Fusarium; Lactation; Lymphocytes; Mycotoxins; Pregnancy; Swine; Trichothecenes; Zearalenone

The Fusarium toxin deoxynivalenol (DON) is of outstanding importance in pig nutrition because of its frequent occurrence in cereal grains at levels high enough to cause adverse effects such as a decrease in feed intake and impairment of the immune system. Thus, simple decontamination procedures would be useful. The present study aimed to examine the effects of wet preservation of triticale contaminated with DON and zearalenone (ZON) with sodium metabisulphite (SBS) on the treatment-related non-toxic derivative of DON (DON-sulfonate, DONS), and on ZON and its metabolites in blood and various physiological specimens of piglets. The uncontaminated control triticale (CON) and the DON-contaminated triticale (FUS) were included in the diets either untreated or SBS treated (CON-SBS, FUS-SBS) and fed to piglets for 28 days starting from weaning. The diet concentrations for DON were 0.156, 0.084, 2.312 and 0.275 mg kg(-1), for DONS were <0.05, <0.05, <0.05 and 1.841 mg kg(-1), and for ZON were <0.001, 0.006, 0.017, and 0.016 mg kg(-1) for each of CON, CON-SBS, FUS and FUS-SBS, respectively. DONS was present in the blood of piglets fed the FUS-SBS at a median concentration of 15.5 ng ml(-1) (3-67 ng ml(-1)), while the median DON concentration amounted to 2 ng ml(-1) (0-5 ng ml(-1)) at the same time. The median DON concentration in the blood of piglets fed the FUS diet reached a median concentration of 10.5 ng ml(-1) (5-17 ng ml(-1)). Moreover, the relative differences between the DON concentrations in other physiological specimens (muscle, liver, kidney, bile and urine) in piglets fed the FUS-SBS and the FUS diet were comparable with the blood DON concentration differences. Although these differences can be taken as an indication for DONS stability after absorption and distribution further studies examining DONS in these other physiological specimens directly are necessary to substantiate this conclusion. Moreover, ZON and α-zearalenol could only be detected in bile and urine where their levels were not influenced by the SBS treatment.

Topics: Administration, Oral; Animal Feed; Animals; Bile; Edible Grain; Female; Food Contamination; Food Preservation; Food Preservatives; Kidney; Liver; Male; Muscle, Skeletal; Sulfites; Swine; Trichothecenes; Weaning; Zearalenone; Zeranol

To elucidate the dietary exposure of Chinese populations to deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN).. Ten Fusarium toxins including DON, NIV, ZEN in domestic wheat flour and corn-based products harvested and collected in 2009 were analyzed by UPLC-MS/MS. Dietary intake assessments of human exposure to DON, NIV and ZEN were carried out in combination of national food consumption data with toxin concentration data by deterministic estimate method.. (1) There are 2.5% adults and 10% children with the dietary exposure to DON exceeding the tolerable daily intake (TDI) on the basis of the average food consumption. At the 75th percentile food consumption level, the dietary exposure of populations to DON was higher than its TDI, 1.72 and 2.02 times (adults) as well as 1.19 and 1.09 times higher than TDI (children), respectively, based on the higher DON exposure (adults : P90 for wheat flour and P97. 5 for corn-based products, children: P50 for wheat flour and P75 for corn-based products). At the average toxin concentration, children with the high consumption level (90th, 97. 5th and 99th percentile) of either wheat flour or corn-based products, the dietary exposure to DON exceed the TDI, 1.81 to 3.17 times (wheat flour) and 1.47 - 3.97 times (corn-based products) higher than TDI, respectively. The dietary exposure of adults to ZEN exceed the TDI, based on the average food consumption data and higher ZEN level (P99), the 75th consumption data combined with P97. 5th toxin concentration, as well as 90th percentile of both food consumption data and ZEN concentration, respectively. There are 1%, 2.5%, 25%, 25% and 50% populations of children exposed ZEN higher than TDI, respectively at the 50th, 75th, 90th, 97. 5th and 99th percentile of food consumption data. (2) The level of concern (LOC) of DON in wheat flour calculated with high consumption data (90th, 97. 5th and 99th percentile) for all populations as well as DON and ZEN in corn-based products calculated with high consumption data (adults: 99th percentile, children: 97. 5th and 99th percentile) were lower than their average concentrations in above cereals. (3) No matter adults or children, the maximum daily safe intake of both wheat flour and corn-based products with high concentration of DON (97. 5th and 99th percentile, and 90th else for children) were lower than their own average food consumption data. For adults, the maximum daily safe intake of corn-based products with high ZEN contamination level (99th percentile) was lower than the average amount of corn-based products consumption. While, the maximum daily safe intake of both wheat flour and corn-based products derived from high ZEN concentration (97.5th and 99th percentile) for children were lower than their own average food consumption.. Children dietary exposure to DON, NIV and ZEN was higher than adults. Children are the populations at the high risk of dietary exposure to these three mycotoxins. The risk to health caused by long-term consumption of wheat flour and corn-based products heavily contaminated with such high concentration of DON and ZEN was relatively high. The present tolerance limit for DON and ZEN in foods implemented in China should be revised on the basis of assessment results.

Topics: Adult; Child; China; Diet; Edible Grain; Environmental Exposure; Food Contamination; Food Microbiology; Fusarium; Humans; Mycotoxins; T-2 Toxin; Trichothecenes; Zearalenone

Nine gilts weighing 80 kg at the beginning of the trial were fed a mycotoxin contaminated diet containing 2 mg deoxynivalenol (DON) and 0.4 mg zearalenone (ZON) per kg (Diet M). Their daily weight gain until 103 kg BW was reduced in comparison to the nine control animals fed an uncontaminated diet (Diet C) (763 vs. 912 g; p = 0.02). There was no treatment effect on the age at first observed oestrus. Seven and eight gilts receiving Diet M and C, respectively, became pregnant after being mated once or being again mated three weeks later. The examination of the uteri of gilts slaughtered 35-61 days after mating showed that the exposure to DON and ZON had no effect on the number of foetuses per gilt (p = 0.54), but increased their growth rate (p = 0.003). Thus, low dietary DON and ZON levels had no negative effects on the reproductive parameters examined. The hypothesis that the bulbourethral gland weight of barrows can be used for the bioassay of low dietary ZON levels was rejected since feeding Diet M from 80-103 kg BW did not increase the weight of that accessory sex gland (p = 0.51).

Topics: Animals; Bulbourethral Glands; Estrogens, Non-Steroidal; Estrus; Female; Fertility; Food Contamination; Litter Size; Male; Organ Size; Pregnancy; Pregnancy Rate; Random Allocation; Swine; Trichothecenes; Weight Gain; Zearalenone

The interest in holistic considerations in the area of food safety is increasing. Risk managers may face the problem that reducing the risk of one compound may increase the risk of another compound. An example is the potential increase in mycotoxin levels due to a reduced use of fungicides in crop production. The Integrated Probabilistic Risk Assessment (IPRA) model was used to compare the estimated health impacts on humans caused by crops contaminated with the fungicides spiroxamine (SPI) and tebuconazole (TEB) or with the mycotoxins deoxynivalenol (DON) and zearalenone (ZEA). The IPRA model integrates a distribution characterising the exposure of individuals with a distribution characterising the susceptibility of individuals towards toxic effects. Its outcome, a distribution of Individual Margins of Exposure (IMoE), served as basis to perform comparisons of compounds, effects, countries, and population groups. Based on the available data and the assumptions made, none of the four compounds was found to have impact on human health in the addressed scenarios. The IMoE distributions were located as follows: DON

Topics: Food Microbiology; Fungicides, Industrial; Humans; Models, Statistical; Mycotoxins; Risk Assessment; Risk Reduction Behavior; Spiro Compounds; Triazoles; Trichothecenes; Zearalenone

Mycotoxins are secondary metabolites produced by microfungi that are capable of causing disease and death in humans and other animals. This work was aimed at investigation of influence of mouldy wheat contaminated by pathogenic fungi producing mycotoxins on metallothionein levels in hepatic tissue of rats. The rats were administrating feed mixtures with different contents of vitamins or naturally mouldy wheat for 28 days. It was found that the wheat contained deoxynivalenol (80 +/- 5 microg per kg of mouldy wheat), zearalenone (56 +/- 3 microg/kg), T2-toxin (20 +/- 2 microg/kg) and aflatoxins as a sum of B1, B2, G1 and G2 (3.9 +/- 0.2 microg/kg). Rats were fed diets containing 0, 33, 66 and 100% naturally moulded wheat. Control group 0, 33, 66 and 100% contained vitamins according to Nutrient Requirements of Rats (NRC). Other four groups (control group with vitamins, vit33, vit66 and vit100%) were fed on the same levels of mouldy wheat, also vitamins at levels 100% higher than the previous mixtures. We determined weight, feed conversion and performed dissection to observe pathological processes. Changes between control group and experimental groups exposed to influence of mouldy wheat and experimental groups supplemented by higher concentration of vitamins and mouldy wheat were not observed. Livers were sampled and did not demonstrate significant changes in morphology compared to control either. In the following experiments the levels of metallothionein as a marker of oxidative stress was determined. We observed a quite surprising trend in metallothionein levels in animals supplemented with increased concentration of vitamins. Its level enhanced with increasing content of mouldy wheat. It was possible to determine a statistically significant decline (p<0.05) between control group and groups of animals fed with 33, 66 and 100% mouldy wheat. It is likely that some mycotoxins presented in mouldy wheat are able to block the mechanism of metallothionein synthesis.

Topics: Aflatoxins; Animals; Body Weight; Fungi; Liver; Male; Metallothionein; Oxidative Stress; Rats; Rats, Wistar; T-2 Toxin; Trichothecenes; Triticum; Vitamins; Zearalenone

Cereals and cereal-based food have often been found to be contaminated with the mycotoxins deoxynivalenol (DON) and zearalenone (ZON), after infection of the grain with the pathogenic fungus Fusarium. Both the pathogen and the infected plants can chemically modify DON and ZON, including acetylation, glucosidation, and sulfation. Analytical strategies for detection and quantification of DON and ZON are well known and established but often fail to recognize the respective metabolites, which are, therefore, also referred to as "masked" mycotoxins. However, several masked forms are also known to be harmful to mammals. Failure to detect these could lead to significant underestimation of the toxic potential of a particular sample. To monitor the levels of DON and ZON metabolites in cereals and cereal-based food, we have developed a LC-MS-MS method capable of simultaneous determination of DON, ZON, and eight of their masked metabolites, namely deoxynivalenol-3-glucoside (D3G), 3-acetyl-deoxynivalenol (3ADON), zearalenone-4-glucoside (Z4G), alpha-zearalenol (alpha-ZOL), beta-zearalenol (beta-ZOL), alpha-zearalenol-4-glucoside (alpha-ZG), beta-zearalenol-4-glucoside (beta-ZG), and zearalenone-4-sulfate (Z4S). The suitability of several cleanup strategies including C(18)-SPE, primary and secondary amines (PSA), MycoSep push-through columns, and immunoaffinity columns was evaluated. The final method used no sample cleanup and was successfully validated for four cereal-based food matrices, namely cornflour, porridge, beer, and pasta, showing good recoveries and precision for all analytes.

Topics: Chromatography, Liquid; Edible Grain; Food Contamination; Solid Phase Extraction; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

The paper describes a sample clean-up method for the co-isolation of deoxynivalenol (DON) and zearalenone (ZON), two mycotoxins naturally co-occurring in wheat. The method is based on immunoaffinity columns prepared by co-immobilising anti-DON and anti-ZON antibodies in a porous sol-gel glass. The main task in developing the method consisted in finding a loading medium allowing retention of both analytes as well as a common elution medium for the dissociation of both antigen-antibody complexes formed. This can be achieved by co-extracting DON and ZON with ACN-water (60:40, v/v), reducing the acetonitril concentration to 2.5% before loading an aliquot of the diluted sample extract onto the DON/ZON column. The columns are washed with 5 ml of MeOH-water (10:90, v/v) before DON and ZON are co-eluted with 4 ml of ACN-water (50:50, v/v). Concentrations of DON and ZON are determined with HPLC-UV and HPLC-fluorescence detection, respectively. The sample clean-up method was shown to be applicable to wheat and wheat products, e.g., cornflakes, milk wheat mash and rusk. Spiking experiments (spike level 500 microg DON/kg and 50 microg ZON/kg) resulted in recovery rates from 82% to 111%.

Topics: Antibodies; Chromatography, Affinity; Food Analysis; Linear Models; Phase Transition; Reproducibility of Results; Sensitivity and Specificity; Trichothecenes; Triticum; Zearalenone

This study examined a total of 82 consignments of French and Argentinean raw maize as received at maize mills in the UK between 2004 and 2007. Samples were analysed for deoxynivalenol (DON), nivalenol (NIV), other trichothecenes, zearalenone (ZON), and fumonisins B(1), B(2), and B(3) (FB(1), FB(2), and FB(3)) using fully validated analytical methods with limits of quantification of 10 microg kg(-1) for DON, NIV, and each fumonisin mycotoxin and 3 microg kg(-1) for ZON. All samples except two containing fumonisins met the European Commission statutory maximum permissible levels for DON, ZON, and FB(1) + FB(2) as operating in 2007. The maximum concentrations found for DON, NIV, ZON, and FB(1) + FB(2) were 444, 496, 165 and 5002 microg kg(-1), respectively. Fumonisins were detected in almost every sample with 65% of Argentinean maize containing more than 1000 microg kg(-1) of FB(1) + FB(2). In contrast, ZON was not detectable in almost 50% of consignments. During this period there was a distinct difference in mycotoxin concentrations between harvests and geographic origin. Flint maize from Argentina usually contained lower concentrations of DON and related trichothecenes and higher levels of fumonisins than maize from France, although concentrations of fumonisins up to 2000 microg kg(-1) or greater occurred in samples from both regions. The incidence and concentrations of fumonisins were similar to those in a similar previous survey, while zearalenone concentrations were lower. The distribution of mycotoxins in multi-hold ships was also investigated showing that fumonisins were much more evenly distributed than DON, thus indicating their general level in the ship as a whole. The effect of cleaning regimes was found to be very variable, especially for DON, ranging from no removal of mycotoxins to greater than 50% in some instances, but was not related to concentration. Evidence here suggests that while cleaning is essential for removing foreign bodies before milling, it cannot be used as a reliable tool for reducing mycotoxins.

Topics: Animal Feed; Animals; Argentina; Food Contamination; Food Handling; France; Fumonisins; Fusarium; Humans; Mycotoxins; Trichothecenes; United Kingdom; Zea mays; Zearalenone

Wheat (as bran) and corn (as dry grain or fermented feed) are main ingredients of feedstuffs used in local cattle and pig farms in the South of the Buenos Aires Province (Argentina). Therefore, determining mycobiota and mycotoxins in wheat and corn is of prime importance for developing feed management techniques to optimise animal production and to minimize toxicity. Then, a mycological survey was carried out in the Southeastern part of the Buenos Aires Province, in order to identify the mycobiota and the main mycotoxins present in fermented feed, wheat grain and corn grain samples. Samples were cultured for fungal quantification, isolation and identification, and analysed for deoxynivalenol (DON), zearalenone (ZEA), T-2 toxin and aflatoxins (AFLA). Penicillium (74%), Aspergillus (32%) and Scopulariopsis (21%) were the prevalent genera in fermented feed. Penicillium (70%), Fusarium (47%) and Aspergillus (34%) were the most frequent fungi isolated from corn. Penicillium (42%), Fusarium (27%) and Alternaria (25%) were the most frequently recovered genera from wheat. DON was detected in 59% of the corn samples, in 45% of the wheat samples and in 38% of the silage samples. ZEA was detected in 36% of the corn samples, in 49% of the wheat samples and in 16% of the silage samples. T-2 toxin and aflatoxin B1 were each detected in 4% of the corn samples. Eighteen percent of the fermented feed samples showed T-2 contamination. Fermented feed and wheat samples were negative for AFLA.

Topics: Aflatoxin B1; Animals; Argentina; Cattle; Fermentation; Food Contamination; Food Microbiology; Fungi; Mycotoxins; Silage; Species Specificity; Swine; Trichothecenes; Triticum; Zea mays; Zearalenone

Contamination by microscopic fungi and mycotoxins in high moisture corn (HMC) silages conserved by chemical additives was investigated. The samples were examined for the concentration and identification of microscopic fungi able to grow on Malt and Czapek-Dox agar and for mycotoxins content (deoxynivalenol, T-2 toxin, zearalenone and total aflatoxins, fumonisins, ochratoxins) by direct competitive enzyme-linked immunosorbent assays. The average fungal counts were 3.37 +- 2.52 log cfu/g in control HMC silages, 2.91 +- 0.51 log cfu/g in HMC silages treated by organic salts and inorganic salt, 3.62 +- 1.46 log cfu/g in HMC ensiled with organic acids and 3.49 +- 1.12 log cfu/g of HMC silages treated by organic acids along with organic salt. In this study, 740 isolates belonging to 10 fungal species representing 9 genera were recovered. The genera of microscopic fungi most frequently found in HMC were Penicillium (56.49 percentages) and Paecilomyces (32.16 percentages). T-2 toxin was the secondary metabolite with the highest concentration ranging from 179.13 +- 3.04 to 249.40 +- 24.69 micrograms/kg, followed by deoxynivalenol and total fumonisins. The highest mean of deoxynivalenol level was 0.13 +- 0.02 mg/kg and concentration of total fumonisins ranged from 20.13 +- 2.53 to 90.33 +- 10.35 micrograms/kg. This study indicated that application of chemical additives containing organic acids, organic salts and inorganic salt was sufficient to inhibit mycotoxins formation. The use of calcium formiate, sodium benzoate and sodium nitrite resulted in high hygienic quality of HMC silages and significantly reduced the concentration of zearalenone, deoxynivalenol and total ochratoxins and fumonisins.

Topics: Colony Count, Microbial; Food Additives; Fumonisins; Fungi; Mycotoxins; Trichothecenes; Zea mays; Zearalenone

The occurrence of mycotoxins in 140 maize silages, 120 grass silages and 30 wheat silages produced in the Netherlands between 2002 and 2004 was determined using a liquid chromatography coupled with tandem mass spectrometry detection (LC-MS/MS) multi-method. Deoxynivalenol (DON) was detected above the limit of quantification (LOQ) of 250 μg kg⁻¹ in 72% of maize and 10% of wheat silages. Average DON concentrations were 854 and 621 μg kg⁻¹, respectively, and maximum concentrations 3142 and 1165 μg kg⁻¹, respectively. Zearalenone was detected above the LOQ of 25 μg kg⁻¹ in 49% of maize and 6% of grass silages. Average zearalenone concentrations were 174 and 93 μg kg⁻¹, respectively, and maximum concentrations 943 and 308 μg kg⁻¹, respectively. The incidences and average concentrations of DON and zearalenone in maize silage were highest in 2004. The incidence of other mycotoxins was low: fumonisin B1 and 15-acetyl-DON were detected in 1.4 and 5% of maize silages, respectively, and roquefortin C in 0.8% of grass silages. None of the silages contained aflatoxins, ochratoxin A, T2-toxin, HT2-toxin, sterigmatocystin, diacetoxyscirpenol, fusarenon-X, ergotamine, penicillinic acid, or mycophenolic acid. This study demonstrates that maize silage is an important source of DON and zearalenone in the diet of dairy cattle. Since the carryover of these mycotoxins into milk is negligible, their occurrence in feed is not considered to be of significant concern with respect to the safety of dairy products for consumers. Potential implications for animal health are discussed.

Topics: Animals; Cattle; Chromatography, High Pressure Liquid; Dairying; European Union; Food Contamination; Food Inspection; Guideline Adherence; Limit of Detection; Mycotoxins; Netherlands; Poaceae; Poisons; Silage; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Trichothecenes; Triticum; Zea mays; Zearalenone

A total of 307-samples of plant crops (both home-grown and imported) marketed in Portugal, including maize and maize-based products (132), wheat and wheat-based products (82), barley (25), soybean (53), sunflower (8) and alfalfa (7), were analyzed for the co-occurrence of the Fusarium toxins, zearalenone (ZEA) and deoxynivalenol (DON). Analysis for ZEA and DON was carried out by HPLC, with fluorescence and UV detection, respectively. Of the 307 samples, ZEA and DON co-occurred in 46 samples (co-occurrence of 15?); 84 samples were DON-positive and 171 were ZEA-positive. The mean ZEA and DON concentrations were 0.17 and 0.07 mg kg(-1), respectively. The highest levels found in the samples were 0.93 mg kg(-1) for ZEA and 17.9 mg kg(-1) for DON. One DON-positive and 13 ZEA-positive samples had contamination levels above the EU maximum legal limit.

Topics: Chromatography, High Pressure Liquid; Crops, Agricultural; Diet; Environmental Exposure; Food Contamination; Fusarium; Hordeum; Humans; Portugal; Trichothecenes; Triticum; Zea mays; Zearalenone

Pregnant sows were fed a control diet (CON, 0.15 mg deoxynivalenol (DON) and 0.0035 mg zearalenone (ZON) per kg diet) or diet containing 15% of Fusarium toxin contaminated triticale (MYCO, 4.42 mg DON and 0.048 mg ZON per kg diet) during days 35-70 of gestation. All sows were fed in a restricted feeding regimen with the same amount of feed (2000 g/d) over the whole study. At the end of the experiment, fetuses were delivered by Caesarian section and samples of spleen and liver of euthanized sows and fetuses were analyzed. At terminal necropsy, no macroscopic lesion was observed in any organ of either sows or fetuses. The histopathological data indicated significant alteration only in elevated iron staining in the red pulp of spleens in sows of MYCO group after 35 days of feeding. The presence of hemosiderin particles in the spleen sections was confirmed by transmission electron microscopical investigation and by an enhanced Fe2+ concentration in spleen. A glycogen increase (p<0.05) was found in liver cells of fetuses in the experimental group. Together, the results provide evidence of spleen dysfunction (hemosiderosis) in sows fed a Fusarium toxin-contaminated wheat, however, with absence of clinical signs. Enhanced glycogen and an impairment of mitochondria in liver of fetuses was present when their mothers consumed the MYCO diet.

Topics: Animal Feed; Animals; Diet; Female; Fetal Development; Fetus; Food Contamination; Fusarium; Glycogen; Hemosiderin; Hemosiderosis; Hepatocytes; Liver; Maternal Exposure; Maternal-Fetal Exchange; Mitochondria, Liver; Pregnancy; Spleen; Swine; Trichothecenes; Zearalenone

To investigate possible co-occurrences of type B trichothecenes and zearalenone within a Fusarium culmorum-infected wheat harvest lot, kernels were fractionated into six groups by visual criteria. The Fusarium-damaged kernels were subdivided into white, shrunken, and red kernel groups, and the remaining kernels were sorted into healthy, black spotted, and nonspecific groups. The distribution patterns of nivalenol, deoxynivalenol, zearalenone, and ergosterol were determined for possible correlations. Significant correlations between the distribution patterns were found for the mycotoxins and ergosterol for the grouped kernels (r = 0.997-0.999, p < 0.0001). Additionally, remarkably outstanding levels of nivalenol (24-fold more than the mean at 1.16 mg/kg), deoxynivalenol (27-fold more than the mean at 0.16 mg/kg), zearalenone (25-fold more than the mean at 77 microg/kg), and ergosterol (17-fold more than the mean at 13.4 mg/kg) were found in the red kernel group. Further, detailed mycotoxin and ergosterol analyses were carried out on various segments (kernel surface, conidia, bran, and flour) of the red kernels. However, the mycotoxin and ergosterol distribution profiles revealed nonsignificant correlations for these kernel segments, with the exception of deoxynivalenol and nivalenol, which were moderately correlated (r = 0.948, p = 0.035).

Topics: Ergosterol; Fusarium; Mycotoxins; Plant Diseases; Seeds; Trichothecenes; Triticum; Zearalenone

This study aimed to discover potential biomarkers for dioxynivalenol (DON) intoxication. B6C3F1 male mice were orally exposed to 0.83, 2.5 and 7.5 mg/kg body weight (bw) DON for 8 days and the differential protein expressions in their blood plasma were determined by SELDI - Time-of-Flight/Mass Spectrometry (TOF/MS) and the immunoglobulins (Igs) G, A, M and E in the serum were investigated. 11.7 kDa protein was significantly highly expressed according to DON administration and this protein was purified by employing a methyl ceramic HyperD F column with using optimization buffer for adsorption and desorption. The purified protein was identified as a haptoglobin precursor by peptide mapping with using LC/Q-TOF/MS and MALDI-TOF/MS and this was confirmed by western blotting and ELISA. IgG and IgM in serum were decreased in a dose-dependent manner and IgA was decreased at 7.5 mg/kg bw DON administration, but the IgE level was not changed. To compare the expressions of haptoglobin and the Igs patterns between aflatoxin B1 (AFB1), zearalenone (ZEA) and DON intoxications, rats were orally administered with AFB1 1.0, ZEA 240 and DON 7.5 mg/kg bw for 8 days. Haptoglobin was increased only at DON 7.5 mg/kg bw, while it was slightly decreased at ZEA 240 mg/kg bw and it was not detected at all at AFB1 1.0 mg/kg bw. IgG and IgA were decreased by DON, but IgG, IgA, IgM and IgE were all increased by AFB1. No changes were observed by ZEA administration. These results show that plasma haptoglobin could be a diagnostic biomarker for DON intoxication when this is combined with examining the serum Igs.

Topics: Aflatoxin B1; Animals; Blood Proteins; Enzyme-Linked Immunosorbent Assay; Haptoglobins; Immunoglobulins; Male; Mass Spectrometry; Mice; Mice, Inbred Strains; Rats; Rats, Wistar; Trichothecenes; Zearalenone

Effects of dietary contamination with various levels of deoxynivalenol (DON) and zearalenone (ZEA) were investigated on Ross 308 hybrid broilers of both sexes. After hatching, all chickens were fed an identical control diet for two weeks. Then chickens of Group 1 received a diet contaminated with DON and ZEA, both being 3.4 mg kg(-1), while Group 2 received DON and ZEA at 8.2 and 8.3 mg kg(-1), respectively. The diet of the control group contained background levels of mycotoxins. Samples of blood and tissues were collected after two weeks. Intake of both contaminated diets resulted in a significantly decreased activity of glutathione peroxidase (GPx) and increased level of malondialdehyde (MDA) in liver tissue, while in kidneys the concentration of MDA was significantly increased only in Group 1. On the other hand, activities of blood GPx and plasma gamma-glutamyltransferase (GGT) were elevated in Group 2 only. Activities of thioredoxin reductase in liver and GPx in duodenal mucosa tissues, superoxide dismutase (SOD) in erythrocytes as well as levels of MDA in duodenal mucosa and alpha-tocopherol in plasma were not affected by dietary mycotoxins. Blood phagocytic activity was significantly depressed in Group 1 and 2. These results demonstrate that diets contaminated with DON and ZEA at medium levels are already able to induce oxidative stress and compromise the blood phagocytic activity in fattening chickens.

Topics: Animal Feed; Animals; Animals, Newborn; Chickens; Dose-Response Relationship, Drug; Female; Food Contamination; Fusarium; Glutathione Peroxidase; Kidney; Lipid Peroxidation; Liver; Male; Malondialdehyde; Oxidative Stress; Phagocytosis; Random Allocation; Trichothecenes; Zea mays; Zearalenone

Ultraviolet (UV) germicidal irradiation has been applied to the sterilization of agricultural products including stored grain for foodstuffs or animal feed. Although UV treatment is known to be effective for killing pathogenic moulds that contaminate the surface of grain, it remains unclear how and to what extent such irradiation is able to eliminate mycotoxins, the fungal metabolites that have adverse effects on human and animal health. We evaluated in vitro the effects of mild (intensity = 0.1 mW cm(-2) at 254 nm UV-C) and strong (24 mW cm(-2)) UV irradiation on two feed-contaminating mycotoxins, zearalenone (ZEN) and deoxynivalenol (DON). When exposed to mild irradiation, the levels of ZEN and DON (both 30 mg kg(-1) initially) were reduced as irradiation time increased, and became undetectable at 60 min. Strong UV irradiation also reduced the mycotoxin levels in the same time-dependent manner, but more rapidly. It was therefore confirmed in vitro that UV irradiation is effective at reducing the levels of ZEN and DON. The present study provides preliminary data for establishing a practical method of using UV irradiation to reduce mycotoxin contamination in grain intended for use in feed.

Topics: Animal Feed; Animals; Dose-Response Relationship, Radiation; Food Contamination; Fungi; Mycotoxins; Trichothecenes; Ultraviolet Rays; Water; Zearalenone

The insulin-like growth factor (IGF) system regulates foetal and placental growth. It can be influenced by dietary factors, but little is known about the effects of mycotoxin feeding on hepatic levels of the IGF system. The aim was (1) to determine the normal foetal and maternal hepatic transcription of major IGF genes at distinct stages of pregnancy and (2) to find out if the mycotoxins deoxynivalenol (DON) and zearalenone (ZON) affect transcript concentrations. Pregnant sows were fed with naturally contaminated diets from days 35 to 70 (experiment 1), or from days 75 to 110 (experiment 2), or with control diets. Foetal hepatic IGF transcripts were determined at days 70 and 110, maternal transcripts at day 70 by quantitative polymerase chain reaction (qPCR). The highest levels of IGF-I transcripts were found in sows. On the contrary, IGF-II was predominantly expressed in foetuses at day 70. Expression of IGF-IR and IGFBP-3 decreased, whereas that of IGFBP-2 increased towards term. IGF-IIR and IGFBP-1 expression was similar in both foetuses and full-term piglets. IGF-IIR showed reduced levels in the maternal liver. The exposure of pregnant sows to DON- and ZON-contaminated diets significantly reduced the maternal and tended to reduce the weight gain of piglets in experiment 2, but had no effects on hepatic levels of IGF transcripts in both experiments. This suggests that mycotoxin-contaminated diets can induce growth depression in pigs during pregnancy without affecting hepatic transcription of major IGF genes.

Topics: Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Diet; Female; Fetus; Food Contamination; Liver; Maternal-Fetal Exchange; Mycotoxins; Pregnancy; Somatomedins; Sus scrofa; Transcription, Genetic; Trichothecenes; Zearalenone

Extrusion technology is used widely in the manufacture of a range of breakfast cereals and snacks for human consumption and animal feeds. To minimise consumer exposure to mycotoxins, the levels of deoxynivalenol (DON) and zearalenone (ZON) in cereals/cereal products and fumonisins B(1) and B(2) (FB(1) and FB(2)) in maize are controlled by European Union legislation. Relatively few studies, however, have examined the loss of Fusarium mycotoxins during processing. The behaviour of FB(1), FB(2) and fumonisin B(3) (FB(3)), DON and ZON during extrusion of naturally contaminated maize flour and maize grits is examined using pilot-scale equipment. DON and ZON are relatively stable during extrusion cooking but the fumonisins are lost to varying degrees. There is some loss of ZON when present in low concentrations and extruded at higher moisture contents. The presence of additives, such as reducing sugars and sodium chloride, can also affect mycotoxin levels. Moisture content of the cereal feed during extrusion is important and has a greater effect than temperature, particularly on the loss of fumonisins at the lower moistures. The effects are complex and not easy to explain, although more energy input to the extruder is required for drier materials. However, on the basis of these studies, the relationship between the concentration of Fusarium toxins in the raw and finished product is toxin- and process-dependent.

Topics: Flour; Food Analysis; Food Contamination; Food Handling; Food Technology; Fusarium; Hot Temperature; Mycotoxins; Trichothecenes; Zea mays; Zearalenone

Wheat contaminated naturally with the Fusarium toxins deoxynivalenol (DON) and zearalenone (ZON) was fed to pregnant Landrace sows for 35days. On day 110, caesarean section was carried out, the offspring were killed immediately after birth, and their livers and spleens examined. At necropsy there were no macroscopic lesions observed in any organ of either sows or piglets. Histopathological evaluation of tissues from sows of the treated group revealed changes in liver and spleen tissues, whereas no significant changes were observed in these tissues in their piglets. Liver damage, as measured by prominent elevated transaminase activities, was not detected in the serum of the sows. In pregnant sows there were individual variations in sensitivity to the Fusarium toxins. In conclusion, it can be assumed that there are no adverse effects on the liver and spleen of full-term piglets when their mothers consumed diets containing up to 9570 and 358mug DON/ZON per kg diet.

Topics: Animals; Animals, Newborn; Ca(2+) Mg(2+)-ATPase; Cell Growth Processes; Female; Fetal Development; Histocytochemistry; Immunoglobulins; Liver; Microscopy, Electron; Mycotoxins; Pregnancy; Spleen; Swine; Trichothecenes; Zearalenone

To determine fungal genera, Aspergillus and Fusarium species and aflatoxin B(1) (AFB(1)), zearalenone (ZEA), deoxynivalenol (DON), fumonisin B(1) (FB(1)) contamination from pre- and postfermented corn silage produced in the most important region of Argentina where silage practice is developed.. Sampling of corn silos was performed manually through silos in transects at three levels: upper, middle and low sections. AFB(1) and FB(1) were quantified by high-performance liquid chromatography, zearalenone by enzyme-linked immunosorbent assay and DON by gas chromatography. Over 90% of the samples showed counts higher than 1 x 10(4) CFU g(-1). Aspergillus flavus and Fusarium verticillioides were the prevalent species. Some tested samples were contaminated with AFB(1), ZEA, DON and FB(1).. This study demonstrates the presence of fungi and AFB(1), ZEA, DON and FB(1) contamination in corn silage in Argentina.. This manuscript makes a contribution to the knowledge of mycotoxins in Argentinean silage in particular because the environmental conditions in this country differ from those of most reports. The comparison of pre- and postfermentation silage is also outstanding. Therefore, information on fungi and mycotoxins present in silage--an increasingly popular commodity--is useful to estimate potential risk for animal and human health.

Topics: Aflatoxin B1; Argentina; Aspergillus flavus; Food Microbiology; Fumonisins; Fungi; Fusarium; Humidity; Hydrogen-Ion Concentration; Mycotoxins; Rain; Silage; Temperature; Trichothecenes; Zea mays; Zearalenone

Deoxynivalenol and zearalenone are among the most prevalent toxins produced by Fusarium spp. They have been investigated in food and feed products for decades but rarely in the environment. We therefore established solid-phase extraction and liquid chromatography-mass spectrometry (LC-MS) methods to quantify these mycotoxins at trace concentrations in aqueous natural samples. In a model emission study, we inoculated a winter wheat field with Fusarium graminearum and subsequently monitored deoxynivalenol and zearalenone in its drainage water. Before during and after harvest in June and July 2007, these toxins were emitted in concentrations from 23 ng/L to 4.9 microg/L for deoxynivalenol and from not detected to 35 ng/L for zearalenone. Simultaneously, in July and August 2007, deoxynivalenol was also detected in a number of Swiss rivers in concentrations up to 22 ng/L and zearalenone was present in several river samples below the method quantification limit. Other mycotoxins might be emitted from Fusarium-infected fields as well, because some of them are produced in similar amounts as deoxynivalenol and zearalenone and exhibit similar or even higher water solubility than deoxynivalenol. The ecotoxicological consequences of the presence of mycotoxins in surface waters remain to be elucidated.

Topics: Chromatography, Liquid; Fusarium; Mycotoxins; Pesticides; Tandem Mass Spectrometry; Trichothecenes; Water Pollutants; Zearalenone

Identification of Fusarium species by traditional methods requires specific skill and experience and there is an increased interest for new molecular methods for identification and quantification of Fusarium from food and feed samples. Real-time PCR with probe technology (Taqman) can be used for the identification and quantification of several species of Fusarium from cereal grain samples. There are several critical steps that need to be considered when establishing a real-time PCR-based method for DNA quantification, including extraction of DNA from the samples. In this study, several DNA extraction methods were evaluated, including the DNeasy Plant Mini Spin Columns (Qiagen), the Bio robot EZ1 (Qiagen) with the DNeasy Blood and Tissue Kit (Qiagen), and the Fast-DNA Spin Kit for Soil (Qbiogene). Parameters such as DNA quality and stability, PCR inhibitors, and PCR efficiency were investigated. Our results showed that all methods gave good PCR efficiency (above 90%) and DNA stability whereas the DNeasy Plant Mini Spin Columns in combination with sonication gave the best results with respect to Fusarium DNA yield. The modified DNeasy Plant Mini Spin protocol was used to analyse 31 wheat samples for the presence of F. graminearum and F. culmorum. The DNA level of F. graminearum could be correlated to the level of DON (r(2) = 0.9) and ZEN (r(2) = 0.6) whereas no correlation was found between F. culmorum and DON/ZEA. This shows that F. graminearum and not F. culmorum, was the main producer of DON in Swedish wheat during 2006.

Topics: DNA, Fungal; Food Microbiology; Fusarium; Mycelium; Mycology; Mycotoxins; Polymerase Chain Reaction; Trichothecenes; Triticum; Zearalenone

In the European Union, deoxynivalenol in cereals and cereal products is controlled by recent legislation with the objective of minimizing consumer exposure to this mycotoxin. Relatively few studies have examined the loss of Fusarium mycotoxins during processing and whether this is accurately reflected by the processing factors. The behaviour of deoxynivalenol, nivalenol and zearalenone during extrusion of naturally contaminated wholemeal wheat flour has been examined using pilot-scale equipment. Factors examined were temperature and moisture content. Concentrations of the three mycotoxins were little changed by extrusion although the amount of deoxynivalenol decreased at the lowest moisture content. However, this effect did not appear to be temperature-dependent, suggesting that the apparent loss is either due to binding or inability to extract the residue. Under some conditions, concentrations of the mycotoxins, particularly nivalenol, were higher after extrusion.

Topics: Edible Grain; Flour; Food Contamination; Food Industry; Fusarium; Humidity; Mycotoxins; Temperature; Trichothecenes; Triticum; Zearalenone

One hundred and fifty-six samples of breakfast cereals were collected from the Canadian retail marketplace over a 3-year period. The samples were analysed for the mycotoxins deoxynivalenol, nivalenol, HT-2 toxin, zearalenone, ochratoxin A, and fumonisins B1 and B2 to contribute to dietary exposure estimates in support of the development of Canadian guidelines for selected mycotoxins in foods. The samples included corn-, oat-, wheat- and rice-based cereals, as well as mixed-grain cereals, and were primarily from North American processors. Overall, deoxynivalenol was the most frequently detected mycotoxin--it was detected in over 40% of all samples analysed. Fumonisins and ochratoxin A were each detected in over 30% of all samples. Zearalenone was detected in over 20% of all samples. Nivalenol and HT-2 toxin were each detected in only one sample. The survey clearly demonstrated regular occurrence of low levels of multiple mycotoxins in breakfast cereals on the Canadian market.

Topics: Canada; Edible Grain; Environmental Monitoring; Food Contamination; Fumonisins; Fusarium; Mycotoxins; Ochratoxins; T-2 Toxin; Trichothecenes; Zearalenone

This study investigates the occurrence of aflatoxins in Ecuador. Early investigators proved the presence of aflatoxins in human and animal food, but the disturbing data lead to the formation of two research teams at Guayaquil University and the Agrarian University of Ecuador to investigate aflaxotins and other mycotoxins in food and their relationship to human health. Because the concept of mycotoxicosis as a result of the secondary metabolites produced by different species of moulds could cause different clinical patterns, the research team includes Aspergillus metabolites found in the urine of a patient with pulmonary aspergilloma. We considered that the body itself could create secondary metabolites. An ELISA method was used to detect mycotoxins with the specific reactive compounds using a company base assay. This allows the detection quantitative of such metabolites in 24 h collected urine. The patient was treated with itraconazole for nine months, after clinical, radiological and aflatoxins testing. We also investigated three other cases in children with a second level of malnutrition and only with vomitoxins results and in three investigated cases of otomycosis caused by Aspergillus niger only in one case traces of aflatoxins were found.

Topics: Adult; Aflatoxins; Aspergillosis; Child; Child, Preschool; Ecuador; Female; Food Contamination; Gastritis; Humans; Lung Diseases, Fungal; Male; Malnutrition; Middle Aged; Mycotoxicosis; Mycotoxins; Otitis Media; Rural Population; Trichothecenes; Zearalenone

Deoxynivalenol, zearalenone, and aflatoxin concentrations in Sicilian durum wheat were determined through ELISA tests. The results highlighted the safety of the grain samples at harvest because deoxynivalenol, zearalenone, and aflatoxin levels did not exceed European legal limits. With regard to aflatoxins, reliability of the ELISA test was evaluated, comparing it with HPLC analyses. The comparison of HPLC and ELISA data showed a tendency to overestimate aflatoxin concentrations with respect to chromatographic determinations. The usefulness of ELISA was confirmed as a rapid screening method; however, when contamination levels are close to legal limits, chromatographic analyses are necessary to quantify aflatoxins with greater accuracy and specificity.

Topics: Aflatoxins; Chromatography, High Pressure Liquid; Edible Grain; Enzyme-Linked Immunosorbent Assay; Europe; Food Contamination; Mycotoxins; Safety; Seasons; Seeds; Sicily; Trichothecenes; Triticum; Weather; Zearalenone

Under unfavorable climatic conditions, Fusarium spp. can contaminate corn plants in the field and produce toxins that are present at the time of ensiling. The stability of deoxynivalenol, fumonisins B1 and B2, and zearalenone in corn silage was tested over two consecutive years. Variables studied were corn dry matter (DM) and storage length and temperature. The concentration of all Fusarium toxins decreased upon ensiling ( P < 0.001). Increasing the length of storage and ensiling with low DM resulted in a higher rate of toxin disappearance, particularly for the water soluble toxins deoxynivalenol and fumonisin B1. Toxin disappearance ranged from 50% for zearalenone to 100% for deoxynivalenol. In contrast, temperature did not have any effect on stability ( P > 0.05). These results indicate that low DM at ensiling as well as a prolonged storage could be a practical way to reduce or eliminate some Fusarium toxins in contaminated silages.

Topics: Food Contamination; Food Preservation; Fumonisins; Fusarium; Mycotoxins; Silage; Time Factors; Trichothecenes; Zea mays; Zearalenone

Multiform single chain variable fragments (scFvs) including different length linker scFvs and bispecific scFv were constructed. The linker lengths of 0, 3, 5, 8, 12, and 15 amino acids between V(H) and V(L) of antideoxynivalenol (anti-DON) scFv were used to analyze the affinities of scFvs. The affinity constants of these scFvs increased when the linker was lower than 12 amino acids. The affinity constant would not change when the linker was longer than 12 amino acids. Fusion gene of anti-DON scFv and antizearalenone (anti-ZEN) scFv was also constructed through connection by a short peptide linker DNA to express a bispecific scFv. The affinity constants assay showed that the two scFvs of fusion bispecific scFv remained their own affinity compared to their parental scFvs. Competitive direct enzyme linked immunosorbent assay was used to detect DON and ZEN in contaminated wheat (Triticum aestivum L.) samples, and the results indicated that this bispecific scFv was applicable in DON and ZEN detection. This work confirmed that bispecific scFv could be successfully obtained, and might also have an application in diagnosing fungal infection, and breeding transgenic plants.

Topics: Antibodies; Antibodies, Bispecific; Antibody Affinity; Base Sequence; Cloning, Molecular; Enzyme-Linked Immunosorbent Assay; Escherichia coli; Food Contamination; Gene Expression; Immunoglobulin Variable Region; Peptides; Trichothecenes; Triticum; Zearalenone

We studied the interactive effects of either binary or tertiary mixtures of Fusarium mycotoxins, deoxynivalenol (DON), zearalenone (ZEA), and fumonisin B1 (FB1) on the human intestinal cell line, Caco-2, using the endpoints including malonedialdehyde (MDA) production, inhibition of protein and DNA syntheses, DNA methylation, DNA fragmentation, and cell viability as measured by the neutral red (NR) test. The mixtures of mycotoxins reduce cellular viability in increasing order: [FB1+ZEA]<[FB1+DON]<[ZEA+DON]<[FB1+DON+ZEA] in NR test. Because FB1 antagonizes the effects of estrogenic Zearalenone, FB1 was assayed against estradiol. In NR assay, mixture of FB1 and estradiol and/or ZEA improves Caco-2 cells viability in contrast to individual effects. Mixtures of ZEA or FB1 and DON, display synergistic effects in lipid peroxidation. The ability of the toxins to inhibit DNA synthesis is 45%, 70%, and 43% for 10 microM of ZEA, DON, and FBI, respectively. Their binary mixtures (at 10 microM each), inhibit DNA synthesis by 35%, 62%, and 65%, far less than additive effects. Surprisingly, the tertiary mixture (10 microM each) only inhibits DNA synthesis by 25%. ZEA, DON, and FB1 induce DNA fragmentation individually. However, mixtures of these mycotoxins always damage DNA to a greater extent. Each individual mycotoxin (10 microM) raises the percentage of 5-methylcytosine (m5dC) in DNA from 4.5% to 9%, while the combination does not increase this rate any further. Altogether, the data indicate that mixtures of Fusarium toxins are able to induce lipid peroxidation, DNA damage, DNA fragmentation, DNA methylation, and cytotoxicity in Caco-2 cells, and suggest a potential promoter effect in human intestinal cells.

Topics: Apoptosis; Caco-2 Cells; Cell Survival; DNA Fragmentation; DNA Methylation; Drug Combinations; Drug Interactions; Enterocytes; Fumonisins; Fusarium; Gene Silencing; Humans; Lipid Peroxidation; Malondialdehyde; Mycotoxins; Oxidative Stress; Trichothecenes; Zearalenone

The high prevalence of the Fusarium mycotoxins, deoxynivalenol (DON) and zearalenone (ZON) in animal feeds in mild climatic zones of Europe and North America results in considerable economic losses, as these toxins affect health and productivity particularly of pigs from all age groups. The use of mycotoxin adsorbents as feed additives is one of the most prominent approaches to reduce the risk for mycotoxicoses in farm animals, and to minimise carry-over of mycotoxins from contaminated feeds into foods of animal origin. Successful aflatoxin adsorption by means of different substances (phyllosilicate minerals, zeolites, activated charcoal, synthetic resins or yeast cell-wall-derived products) has been demonstrated in vivo and in vitro. However, attempts to adsorb DON and ZON have been less encouraging. Here we describe the adsorption capacity of a variety of potential binders, including compounds that have not been evaluated before, such as humic acids. All compounds were tested at realistic inclusion levels for their capacity to bind ZON and DON, using an in vitro method that resembles the different pH conditions in the gastro-intestinal tract of pigs. Mycotoxin adsorption was assessed by chemical methods and distinct bioassays, using specific markers of toxicity as endpoints of toxicity in cytological assays. Whereas none of the tested substances was able to bind DON in an appreciable percentage, some of the selected smectite clays, humic substances and yeast-wall derived products efficiently adsorbed ZON (>70%). Binding efficiency was indirectly confirmed by the reduction of toxicity in the in vitro bioassays. In conclusion, the presented test protocol allows the rapid screening of potential mycotoxin binders. Like other in vitro assays, the presented protocol combining chemical and biological assays cannot completely simulate the conditions of the gastro-intestinal tract, and hence in vivo experiments remain mandatory to assess the efficacy of mycotoxin binders under practical conditions.

Topics: Adsorption; Animal Feed; Animals; Cell Line, Tumor; Cell Survival; Charcoal; Chromatography, High Pressure Liquid; Food Additives; Humans; Hydrogen-Ion Concentration; Mice; Molecular Structure; Mycotoxicosis; NIH 3T3 Cells; Trichothecenes; Zearalenone

Given the prominence and the growing importance of mycotoxins in human and animal health, and particularly of vomitoxin and zearalenone in people who use wheat and wheat products as their staple diet, we investigated two different types of wheat milling. Wheat produced according to good manufacturing practice related to mycotoxin risks (from sowing to harvesting) was used to compare the vomitoxin and zearalenone content of soft wheat flour, following the use of two different types of milling, traditional milling with a stone mill and modern milling with a roller mill. Moreover, the vomitoxin and zearalenone content was also evaluated in commercial stone-milled and roller-milled flours. Our results show that stone milling reduced vomitoxin and zearalenone content in flours, compared with the use of the roller-mill system.

Topics: Consumer Product Safety; Flour; Food Handling; Food Microbiology; Humans; Trichothecenes; Zearalenone

1. Diets with increasing proportions of Fusarium toxin-contaminated wheat (0, 170, 340 and 510 g CW/kg) were fed to male turkeys (BUT Big 6) from d 21 to d 56 of age. Each diet was tested with or without a non-starch-polysaccharide (NSP) hydrolysing enzyme preparation. Dietary deoxynivalenol (DON) and zearalenone (ZON) concentrations were successively increased up to approximately 5.4 and 0.04 mg/kg, respectively. 2. Weight gain decreased slightly with increasing proportions of CW, by 1.6, 0.7 and 3.6%, whereas other performance parameters remained unaffected. NSP enzyme supplements to the diets had no influence. 3. The weight of the emptied jejunum plus ileum, relative to live weight, decreased in a dose-related fashion whereby the NSP enzyme exerted an additional weight-decreasing effect. A similar weight-decreasing NSP enzyme effect was noted for heart weights. Activity of glutamate dehydrogenase in serum was significantly increased in groups fed the diets with the highest CW proportion, whereas gamma-glutamyl-transferase remained unaltered. 4. Viscosity in the small intestine was significantly reduced by supplementing the diets with the NSP enzyme. This effect successively decreased with increasing proportions of the CW. 5. Concentrations of DON and of its de-epoxidised metabolite de-epoxy-DON in plasma, liver and breast meat were lower than the detection limits of 2 ng/ml (plasma) and 4 ng/g, respectively, of the applied HPLC method. DON concentration in bile reached up to 13 to 23 ng/ml whereas de-epoxy-DON concentration was lower than 4 ng/ml. 6. ZON or its metabolites were not detectable in plasma, liver or breast meat (detection limits of the HPLC method were 1, 0.5 and 5 ng/g for ZON, alpha-zearalenol (ZOL) and beta-ZOL, respectively). Concentrations of ZON and alpha-ZOL in bile increased with dietary ZON concentration. The mean proportions of ZON, alpha-ZOL and beta-ZOL of the sum of all three metabolites were 19, 77 and 4%, respectively.

Topics: Animal Feed; Animals; Dose-Response Relationship, Drug; Eating; Endo-1,4-beta Xylanases; Food Contamination; Fusarium; gamma-Glutamyltransferase; Glutamate Dehydrogenase; Male; Random Allocation; Trichothecenes; Triticum; Turkeys; Weight Gain; Zearalenone

Pregnant sows were fed either a control diet (CON, n=8, 0.21 mg DON and 0.004 mg ZON/kg diet) or a diet containing 40% of a Fusarium toxin contaminated wheat (MYCO, n=7, 9.57 mg DON and 0.358 mg ZON/kg diet) from day 75 to 110 of gestation. Piglets were delivered by Caesarean section at the end. Spleen weights of piglets from the MYCO group were significantly lower. Hemoglobin concentration and hematocrit were also significantly decreased in these piglets, although this effect was more obvious in female than in male piglets. The transfer of DON and ZON was evaluated by the diet ratio (sum of concentrations of all metabolites in the physiological specimen divided by the dietary toxin concentration) and the piglet ratio (sum of concentrations of all metabolites in the physiological specimen of the piglet divided by that of the sows). The diet ratio for the liver (sows only) amounted to 0.001 (DON+de-epoxy-DON) and 0.016 (ZON and metabolites). The diet ratios of DON in bile reached up to 0.041 and 0.003 for sows and piglets, respectively, and those for ZON up to 2.896 and 0.128. The piglet ratios in bile varied up to 0.309 and 0.518 for DON and ZON, respectively, whereas nearly similar DON concentrations were found in serum of piglets and sows (median piglet ratio of 0.750). The results of the study suggest that the developing fetus is exposed to DON, ZON and their metabolites when the sows are fed a Fusarium toxin contaminated diet.

Topics: Animal Nutritional Physiological Phenomena; Animals; Animals, Newborn; Female; Fetus; Food Contamination; Fusarium; Gestational Age; Hematocrit; Hemoglobins; Male; Maternal-Fetal Exchange; Organ Size; Pregnancy; Random Allocation; Sex Factors; Spleen; Swine; Trichothecenes; Zearalenone

The frequent contamination of grain with the Fusarium toxins, deoxynivalenol (DON) and zearalenone (ZON), is an important issue in animal and human nutrition. However, data on the exposure of humans to these toxins through consumption of animal tissues exposed to Fusarium toxins (carry-over) are fragmentary. Therefore, residues of DON, ZON and their metabolites were determined in tissues and body fluids of pigs (female and castrated male) from a fattening trial. Pigs were fed a control (n = 6, 0.24 mg DON and 0.009 mg ZON per kg diet as fed) or a Fusarium toxin-contaminated diet (n = 12, 6.68 mg DON and 0.056 mg ZON per kg diet as fed) either ad libitum or for restrictive consumption for 12 weeks. After slaughter (96.3 +/- 11.6 kg live weight), the concentrations of DON and its metabolite, de-epoxy-DON, were measured in serum, bile, liver, kidney, musculus longissimus and back fat, while ZON and its metabolites, alpha- and beta-zearalenol (alpha-/beta-ZOL), were determined in serum, bile and liver. The mean carry-over factor of DON + de-epoxy-DON, defined as the concentration of both substances in the tissue/fluid divided by the DON concentration in the diet, for all pigs decreased from bile (0.1046 +/- 0.0653) >> kidney (0.0151 +/- 0.0070) > liver (0.0057 +/- 0.0043) > serum (0.0023 +/- 0.0018) > muscle (0.0016 +/- 0.0016) >> back fat (0.0002 +/- 0.0004). The time interval between the end of feeding and slaughter had no consistent effect on DON + de-epoxy-DON concentrations in the analysed specimen of Fusarium toxin-exposed pigs fed restrictively. No transfer of ZON and its metabolites could be observed into serum of pigs, while the mean carry-over factors of ZON + alpha-ZOL + beta-ZOL were 0.0094 +/- 0.0123 and 4.0 +/- 2.2 for liver and bile, respectively. Therefore, it can be concluded that serum is a reliable indicator for DON exposure, but an inappropriate parameter to deduce ZON exposure, which is better represented by bile concentration of ZON + alpha-ZOL + beta-ZOL. However, the exposure risk to humans by consumption of edible tissues of animals exposed to Fusarium toxins is negligible compared to the direct consumption of grain-based food.

Topics: Adipose Tissue; Animal Feed; Animals; Bile; Female; Food Contamination; Fusarium; Kidney; Liver; Male; Muscles; Swine; Trichothecenes; Triticum; Zearalenone

Fermentative bacteria can potentially be utilized to detoxify corn silage contaminated by Fusarium toxins. The objective of the present study was to test a large number of these bacteria for their ability to bind and/or biotransform deoxynivalenol (DON), zearalenone (ZEN) and fumonisins B(1) and B(2) (FB(1), FB(2)) in conditions simulating corn silage. A total of 202 strains were screened in contaminated, pH 4, corn infusion inoculated with 5 x 10(8) CFU ml(-1). Eight Lactobacilli and three Leuconostoc biotransformed ZEN into alpha-zearalenol, but no biotransformation was detected for DON and fumonisins. In contrast, most strains were capable of binding Fusarium toxins. The most effective genera were Streptococcus and Enterococcus, capable of binding up to 33, 49, 24 and 62% of DON, ZEN, FB(1) and FB(2), respectively. The ability to bind Fusarium toxins seems to be a common property of fermentative bacteria and could help to decrease their toxicity in animals.

Topics: Biotransformation; Carcinogens, Environmental; Fermentation; Food Contamination; Food Microbiology; Fumonisins; Fusarium; Gram-Positive Bacteria; Lactobacillus; Lactococcus; Leuconostoc; Silage; Streptococcus; Trichothecenes; Zea mays; Zearalenone

Cell transformation assays using BALB/3T3 cells can mimic the two-stage process of chemical carcinogenesis in experimental animals. A short-term transformation assay using v-Ha-ras-transfected BALB/3T3 cells (Bhas 42 cells), which was developed by Ohmori et al. and modified by Asada et al., has been reported to detect both tumor initiators and promoters as transformation initiators and promoters, respectively, with their differences based on their protocols. In this new short-term assay, we examined mycotoxins derived from Fusarium and related substances for the initiation and promotion activities of the transformation. The tested substances included deoxynivalenol, nivalenol, fusarenon-X, T-2 toxin, fumonisin B(1), fumonisin B(2), zearalenone, alpha-zearalanol, beta-zearalanol, alpha-zearalenol and beta-zearalenol. Fumonisin B(1) and T-2 toxin were positive for promoting activity in the assay. Especially, T-2 toxin was active at concentrations as low as 0.001-0.002microg/mL in the culture medium. From a comparison between the results of this study and published carcinogenicity assay data, it was expected that the Bhas 42 cell transformation assay had a good correlation with the two-stage carcinogenicity tests using experimental animals for estimation of the tumor-promoting activity.

Topics: Animals; BALB 3T3 Cells; Carcinogenicity Tests; Cell Survival; Cell Transformation, Neoplastic; Dose-Response Relationship, Drug; Fumonisins; Fusarium; Genes, ras; Mice; Mycotoxins; T-2 Toxin; Transfection; Trichothecenes; Zearalenone

Eleven pregnant sows with a body weight between 153 and 197 kg were fed a control diet (CON, 0.15 mg DON and 0.0035 mg ZON/kg diet) or a diet containing 15% of Fusarium toxin contaminated triticale (MYCO, 4.42 mg DON and 0.048 mg ZON/kg diet) in the period of day 35 and 70 of gestation. The indirect effect of feed intake was separated from the direct effects of the Fusarium toxins by the restricted feeding regimen where all sows were fed the same amount of feed (2000 g/d) over the whole study. At the end of experiment, fetuses were delivered by Caesarian section and samples of serum, bile, urine, liver, kidney and spleen of euthanatized sows and fetuses were taken to analyze the concentrations of DON, ZON and their metabolites. Feeding the Fusarium toxin contaminated diet to pregnant sows caused neither adverse effects on performance, organ weights and maintenance of pregnancy of sows nor on fetus weight and length. Furthermore, no teratogenic or embryolethal effects could be observed in the MYCO group. Hematological and clinical-chemical parameters of sows and fetuses were not affected by feeding, with the exception of significantly lower GLDH (glutamate dehydrogenase) serum activities in MYCO sows. The carry over of DON and ZON from the diet to the sow or fetus tissues was calculated by the diet ratio (sum of concentrations of all metabolites in the physiological specimen divided by the dietary toxin concentration), while the fetus ratio was evaluated by the sum of concentrations of all metabolites in the physiological specimen of the fetus divided by that of the sows. DON and deepoxy-DON were found in urine, bile, serum, liver, kidney and spleen of sows of the MYCO group, but not in the bile of fetuses (spleen not analyzed). ZON and its metabolite alpha-zearalenol (alpha-ZOL) were detected in urine and bile of sows, while all specimens of fetuses as well as serum and liver of sows were negative for ZON metabolites. The maximum diet ratios for urine and bile in sows of the MYCO group were 0.84 and 0.05 for DON metabolites and 1.2 and 3.8 for ZON metabolites, underscoring the differences in metabolism and excretion of both toxins. The maximum diet ratio of DON and deepoxy-DON into liver, kidney and spleen of MYCO sows were 0.003, 0.007 and 0.003, respectively. The maximum fetus ratio of DON and deepoxy-DON into urine, bile, serum, liver and kidney of fetuses were 0.006, 0, 0.5, 0.88, and 0.33, while the maximum placental ratio (sum of toxin concentration

Topics: Animals; Animals, Newborn; Body Weight; Eating; Estradiol; Estrogens, Non-Steroidal; Female; Fetus; Food Contamination; Fusarium; Gestational Age; Glutamate Dehydrogenase; Male; Maternal-Fetal Exchange; Mycotoxins; Organogenesis; Placental Circulation; Pregnancy; Progesterone; Swine; Trichothecenes; Zearalenone

Among the main Spanish commercially available trademarks, we have selected a total of 25 samples of corn-based foods, which have the highest consume rate, to carry out the analysis of deoxynivalenol (DON), T-2 toxin, zearalenone (ZEA) and zearalenols (ZOL). The contents of mycotoxins were determined by gas chromatography with flame ionization detection, and those of ZEA were confirmed by HPLC with fluorescence detection. Of the 25 analyzed samples, the incidence of DON, ZEA and alfa-ZOL was 68, 44 and 24%, respectively; levels detected ranged from 29-195, 34-216, and 36-71 microg/kg, respectively. T-2 toxin was only detected in one sample (<50 microg/kg). Beta-ZOL was not present in excess of the detection limit in the investigated samples. The results suggest a risk for consumers of corn products and the need to monitor the final products before consumption. This is the first report in Spain on natural contamination with these mycotoxins in corn-based foods.

Topics: Food Contamination; Food Handling; Food Microbiology; Spain; T-2 Toxin; Trichothecenes; Zea mays; Zearalenone; Zeranol

Fungi of the Fusarium species can infect food and feed commodities and produce the mycotoxins zearalenone (ZEA) and deoxynivalenol (DON). Since both toxins have been reported to reduce fertility, the mechanisms of ZEA and DON on inhibition of oocyte maturation were examined. Pig oocytes were matured in the presence of ZEA (a mycotoxin with estrogenlike activity), 17beta-estradiol, and DON (all 3.12 micromol/L). Zearalenone, 17beta-estradiol, and DON inhibited oocyte maturation and caused approximately 34% of the oocytes to form an aberrant spindle. Different ratios of ZEA:DON did not lead to a more severe inhibition of oocyte maturation. Both mycotoxins caused abnormal formation of the meiotic spindle. The developmental competence of oocytes matured in the presence of mycotoxins was further investigated after in vitro fertilization. Presence of ZEA (3.12 micromol/L) during maturation reduced the percentages of oocytes that cleaved and formed a blastocyst to about 12%, compared with 25% of control oocytes. Maturation in the presence of equimolar concentrations of DON was not compatible with development. The ploidy of blastomeres from blastocysts derived from mycotoxin-exposed oocytes was analyzed with fluorescent in situ hybridization. All blastocysts, even those from the control group, contained at least one blastomere with abnormal ploidy, but the variation in the percentages of aneuploid blastomeres was significantly larger in embryos from oocytes exposed to mycotoxins. It is concluded that ZEA and DON can lead to abnormal spindle formation, leading to less fertile oocytes and embryos with abnormal ploidy, and that the effects of ZEA and DON are not synergistic.

Topics: Aneuploidy; Animals; Cell Nucleus; Cells, Cultured; Embryo, Mammalian; Embryonic Development; Estradiol; Fusarium; Mycotoxins; Oocytes; Spindle Apparatus; Swine; Trichothecenes; Zearalenone

Fusarium mycotoxins deoxynivalenol (DON), T-2 toxin, and zearalenone (ZEN) contamination in 5 kinds of cereal grain harvested in 2004 and 2005 in different regions of Lithuania was examined for their occurrence frequency and level. In all cereal species DON was the most frequently detected mycotoxin with an incidence rate of 98.0-100% and range in positive samples from traces to 691 microg kg(-1) in 2004 and 62.5-94.0%, range from traces to 1,121 microg kg(-1) in 2005, respectively. All the tested oat samples collected in 2004-2005 were found to be contaminated with the T-2 toxin. In one sample from the year 2004 the level of T-2 toxin (121.5 microg kg(-1)) exceeded the allowable level. In 2004, ZEN contamination was more frequent in spring wheat, barley and oats grain, whereas in 2005 this toxin was identified at higher levels only in barley grain (68.0%). In one barley grain sample from 2004, ZEN content (193.4 microg kg(-1)) exceeded the allowable level. Variation in the relative air-humidity exerted some effect on the incidence of Fusarium spp. fungi and mycotoxin content in wheat grain. The weather conditions at harvesting contributed to an increase in the contents of Fusarium fungi and DON and ZEN mycotoxins produced by them in winter wheat grain. This risk factor increases the threat to human and animal health.

Topics: Animals; Consumer Product Safety; Edible Grain; Food Contamination; Food Microbiology; Fusarium; Humans; Lithuania; Mycotoxins; Risk Factors; T-2 Toxin; Trichothecenes; Zearalenone

Pearl millet is increasingly being grown as a premium-value grain for the recreational wildlife and poultry industries in the southern US. We conducted three experiments to assess grain mold development in storage conditions typically encountered in the region of production. Variables included production year, temperature, relative humidity, atmosphere, and grain moisture content. In the first experiment, grain was stored for 9 weeks at 20 or 25 degrees C and maintained at 86% or 91% relative humidity (r.h.). In the second experiment, grain was stored for 9 weeks at 20 or 25 degrees C in either air (aerobic) or N2 (anaerobic), and maintained at 100% r.h. In the third experiment, high-moisture grain was stored for 3 weeks at 20 or 25 degrees C and maintained at 100% r.h. Grain was sampled at weekly intervals and plated to determine changes in fungal frequency. Fungi isolated included Fusarium chlamydosporum (19% of grain), Curvularia spp. (14%), F. semitectum (16%), Alternaria spp. (9%), Aspergillus flavus (8%), "Helminthosporium"-type spp. (6%), and F. moniliforme sensu lato (3%). Year of grain production significantly affected isolation frequency of fungi. Isolation frequencies from low-moisture grain were rarely affected by temperature, relative humidity, or atmosphere treatments, but was affected by storage duration for some fungi. Changes in isolation of toxigenic fungi occurred in high-moisture grain. Isolation frequency of F. chlamydosporum increased in grain stored at 86% and 91% r.h. Incidence of A. flavus increased in high-moisture grain treatments, particularly at 25 degrees C. Incidence of deoxynivalenol was not affected by storage treatment. Low concentrations of nivalenol were detected in most grain incubated at 100% r.h. Zearalenone was detected only when grain moisture content was 20-22%. Aflatoxin contamination averaged 174 ng g(-1) over all treatments, and increased up to 798 ng g(-1) in high-moisture grain at stored at 25 degrees C.

Topics: Aerobiosis; Aflatoxins; Anaerobiosis; Ascomycota; Aspergillus; Food Microbiology; Fungi; Fusarium; Humidity; Mycotoxins; Pennisetum; Temperature; Time Factors; Trichothecenes; Zearalenone

A liquid chromatography/tandem mass spectrometry method was developed for the simultaneous determination of aflatoxins (B(1), B(2), G(1), G(2)), ochratoxin A, fumonisins (B(1), B(2)), deoxynivalenol, zearalenone, T-2 and HT-2 toxins in maize. A double extraction approach, using a phosphate-buffered solution followed by methanol, was applied to achieve effective co-extraction of the 11 mycotoxins under investigation having quite different polarities and chemical structures. A new multitoxin immunoaffinity column containing antibodies for all these mycotoxins was used to clean up the extract. Detection and quantification of the 11 mycotoxins were performed by reversed-phase liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry (LC/ESI-MS/MS) using, as chromatographic mobile phase, a linear gradient of methanol/water containing 0.5% acetic acid and 1 mM ammonium acetate. Method performances were quite satisfactory for all tested mycotoxins at contamination levels close to or below the relevant EU maximum permitted or recommended levels. Limits of detection in maize ranged from 0.3 to 4.2 microg/kg. Recoveries higher than 79% were obtained for all tested mycotoxins with relative standard deviations less than 13%.

Topics: Aflatoxins; Chromatography, Affinity; Chromatography, High Pressure Liquid; Food Contamination; Fumonisins; Mycotoxins; Ochratoxins; Spectrometry, Mass, Electrospray Ionization; T-2 Toxin; Tandem Mass Spectrometry; Trichothecenes; Zea mays; Zearalenone

A feeding trial using 220 weaner pigs which comprised two experimental series was conducted to investigate the effects of diets contaminated with the Fusarium toxins deoxynivalenol (DON) and zearalenone (ZON) and to test the hypothesis that apple pomace acts as an antidote to these mycotoxins. Two diets without contaminated wheat, containing either no pomace or 8% pomace, and two diets with naturally contaminated wheat (3.2 mg DON and 0.06 mg ZON, and 2.1 mg DON and 0.25 mg ZON per kg diet in series 1 and 2 respectively), containing either no pomace or 8% pomace were fed ad libitum for 5 weeks. Mycotoxin exposure lowered feed intake (p < 0.01) and growth (p = 0.05), and tended to decrease the energy conversion ratio (p = 0.06). Although the intake of apple pomace did not increase feed intake, it increased the growth rate (p = 0.04), mainly by restoring growth in the presence of mycotoxins (p = 0.08 for the interaction mycotoxin x pomace). In the first experimental series, the animals were immunized with a parvovirus vaccine. The percentage of seroconverting animals did not differ between the treatments (p = 0.56), which indicates that DON did not affect the humoral immune response. In the second experimental series, female piglets fed the contaminated diets had heavier uteri than piglets fed the uncontaminated diets (p < 0.01), regardless of pomace supplementation. The results show that pomace may alleviate the negative effect of DON on growth but does not counteract the hormonal effects of ZON.

Topics: Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Animals, Newborn; Antibody Formation; Dose-Response Relationship, Drug; Eating; Female; Food Contamination; Fusarium; Male; Malus; Organ Size; Random Allocation; Swine; Treatment Outcome; Trichothecenes; Zearalenone

A multianalyte lateral-flow technique using colloidal gold-labeled monoclonal antibodies was developed for the rapid simultaneous detection of deoxynivalenol (DON) and zearalenone (ZEA). The results of this qualitative one-step test were interpreted visually. A very simple and fast sample preparation was used, and the assay procedure could be accomplished within 10 min. When applied to spiked wheat samples, the technique gave accurate and reproducible results. Cut-off levels of 1500 and 100 microg kg(-1) for DON and ZEA, respectively, were observed. The described multianalyte format can be used as a reliable, rapid and cost-effective on-site screening technique for the simultaneous determination of mycotoxins in grain samples.

Topics: Antibodies, Monoclonal; Colloids; Gold; Immunoassay; Time Factors; Trichothecenes; Zearalenone

The objective of the present study was to investigate the effects of Fusarium toxin contaminated wheat and wheat chaff (mycotoxin diet) on nutrient degradability and the metabolism of the mycotoxins deoxynivalenol (DON) and zearalenone (ZON) using the rumen simulations technique (Rusitec). A 6 day application period with control wheat and wheat chaff (control diet) was followed by an 8 day sampling phase. During this time three fermenters received the mycotoxin diet (64.9 mg DON/kg dry matter (DM) and 500 microg ZON/kg DM) and the remaining fermenters served as the controls (1.0mg DON/kg DM and 6 microg ZON/kg DM). Feed residues of the bags and samples of the effluent liquids were pooled per fermenter during the last 8 days of the experiment. Additionally, effluents of the mycotoxin fermenters were taken 6, 12 and 24h after the morning feeding on the first day of the sampling phase. The degradation of organic matter (OM; P<0.05), neutral detergent fibre (NDF; P<0.01) and protein (P<0.001) were increased by administration the Fusarium contaminated diet which was accompanied by an increased ammonia concentration (P<0.01) and increased butyrate (P<0.01), isobutyrate (P<0.01) and isovalerate (P<0.05) values of the mycotoxin effluents in relation to the controls. High proportions of ingested DON of 90% (85-93%) and ingested ZON of 93% (80-104%) were recovered at the pooled feed residues and effluents in form of DON and de-epoxy DON, and ZON and alpha-ZOL after administering the Fusarium toxin contaminated feed. While adsorption of DON as DON and de-epoxy DON in the feed particles was only minor (5%), a higher amount of 38% of ingested ZON was recovered as ZON and alpha-ZOL at the feed residues. The total recovery of DON plus de-epoxy DON in effluents as a percentage of DON intake reached 8%, 9% and 22% of ingested DON at 6, 12 and 24h after application of the contaminated diet the first time, whereby the recovery of de-epoxy DON as percentage of DON intake was only 5% at 24h. Concentrations of ZON and metabolites were lower than detection limits in the time dependent effluent samples.

Topics: Animal Feed; Animals; Fermentation; Food Contamination; Fusarium; Quaternary Ammonium Compounds; Rumen; Sheep; Trichothecenes; Triticum; Zearalenone

Feeding experiments with diets containing Fusarium toxin-contaminated wheat were conducted to clarify the pathogenesis of immunological effects of Fusarium toxins to porcine spleen cells. Contaminated diets were fed to 36 Landrace prepubertal gilts for 35 d. Concentrations (as-fed basis) of the indicator toxins deoxynivalenol (DON) and zearalenone (ZON), respectively, were 210 and 4 (control--group I), 3,070 and 88 (group II), 6,100 and 235 (group III), and 9,570 and 358 microg/kg (group IV). No signs of hyperestrogenism or uterotrophic effects were observed because of dietary treatments. The feeding of mycotoxin-contaminated diets did not cause gross pathological findings in the spleens of animals. In vivo, no inhibitory effects were detected on concanavalin A-stimulation of blood lymphocytes; however, the proliferation rate of splenocytes was inhibited (P < 0.05) in pigs fed the diet with the highest DON/ZON concentration. With in vitro studies, lower proliferation rates of blood lymphocytes and splenocytes preexposed to DON were detected. Serum IgA concentrations of pigs in group II were increased (P < 0.05) compared with the baseline value before feeding the DON/ZON diet. The histopathological data indicated elevated (P < 0.05) iron staining in the red pulp of spleens in gilts from groups I to IV after 35 d of feeding. The presence of hemosiderin particles in the spleen sections was confirmed by transmission electron microscopic investigation. Together, the results provide evidence of spleen dysfunction (hemosiderosis) in the absence of clinical signs, especially in pigs fed higher concentrations (groups III and IV) of Fusarium toxin-contaminated wheat.

Topics: Animal Feed; Animals; Dose-Response Relationship, Drug; Edible Grain; Feeding Behavior; Female; Food Contamination; Fusarium; Mitogens; Spleen; Trichothecenes; Zearalenone

Mycotoxins are contaminants of animal feed that can impair fertility and cause abnormal fetal development in farm animals. The aim of the present study was to investigate the influence of Fusarium-toxin contaminated feed on cumulus morphology and maturation of pig oocytes. Naturally with the Fusarium-toxins deoxynivalenol (DON) and zearalenone (ZON) contaminated wheat was included in feed for gilts at increasing proportions which resulted in increasing dietary concentrations of both toxins (in mgtoxin/kg feed: Group 1 (control), 0.21 and 0.004; Group 2, 3.07 and 0.088; Group 3, 6.1 and 0.235; Group 4, 9.57 and 0.358, for DON and ZON, respectively). Oocytes were recovered from gilt ovaries by follicle aspiration after ovario-hysterectomy. Granulosa cells were analyzed for the expression of the P450(SCC) and 3beta-HSD mRNA by RT-PCR and additionally for P450(SCC) protein by Western blotting. Neither the expression of the P450(SCC) nor of the 3beta-HSD mRNA or the abundance of the P450(SCC) protein was significantly influenced by the mycotoxin application. The distribution of different cumulus cell morphologies was not influenced by group. At the time of recovery, oocytes with compact cumuli in Groups 3 and 4 showed a reduced proportion having immature chromatin in comparison to that for Groups 1 and 2. The proportion of oocytes having degenerated meiotic chromatin was significantly higher in Group 4 than in the other groups. The proportion of oocytes reaching metaphase II in culture was significantly lower in Groups 3 and 4 than in Group 1, and tended to be lower in Group 2 than in Group 1. We conclude that oocyte quality is significantly reduced by feeding of Fusarium-toxins to gilts.

Topics: 3-Hydroxysteroid Dehydrogenases; Animal Feed; Animals; Cattle; Cholesterol Side-Chain Cleavage Enzyme; Chromatin Assembly and Disassembly; Eating; Female; Gene Expression; Male; Meiosis; Oocytes; Oogenesis; Ovarian Follicle; RNA, Messenger; Swine; T-2 Toxin; Trichothecenes; Zearalenone

The aims of this study were to examine the effects of and possible interactions between dry matter (DM) intake and feeding Fusarium toxin-contaminated wheat on ruminal fermentation, serum chemical parameters and milk yield of dairy cows. Fourteen dairy cows equipped with ruminal and duodenal cannulas were analysed. All animals were fed the same ration, the daily feed amounts being adjusted to current performance. On DM basis, the ration consisted of 60% concentrate including 55% wheat [Fusarium-contaminated wheat (mycotoxin period) or control wheat (control period)] and was completed with 40% maize and grass silage. Each cow was fed the contaminated wheat [deoxynivalenol (DON), 8.21 mg/kg DM and zearalenone (ZON), 0.09 mg/kg DM] and the control wheat (0.25 mg DON/kg DM and 51 microg ZON/kg DM). As expected, a higher organic matter (OM) intake decreased the amounts of fermented crude nutrients related to the respective intakes. An increased amount of crude protein degraded (p < 0.05) and a lower molar percentage of propionate in the rumen fluid were observed when feeding the Fusarium toxin-contaminated wheat at increased OM intakes in comparison with the control wheat. The activities of serum aspartate aminotransferase (ASAT; p < 0.001), glutamate dehydrogenase (GLDH; p < 0.01) and gamma glutamyl transferase (gamma-GT; p < 0.01) increased with increasing OM intake and were not related to the mycotoxin contamination of the wheat.

Topics: Animal Feed; Animals; Aspartate Aminotransferases; Blood Chemical Analysis; Cattle; Duodenum; Eating; Energy Intake; Female; Fermentation; Food Contamination; Fusarium; gamma-Glutamyltransferase; Glutamate Dehydrogenase; Milk; Mycotoxins; Random Allocation; Rumen; Trichothecenes; Triticum; Zearalenone

Feeding experiments with diets containing Fusarium toxin-contaminated wheat were conducted to clarify the pathogenesis of enzymatic and histopathological effects of Fusarium toxins on porcine liver cells. A total of 36 prepuberal gilts were divided into four groups and fed diets with increasing proportions of Fusarium toxin-contaminated wheat at a total wheat proportion of 40% over a period of 35 days. The concentrations of the indicator toxins deoxynivalenol (DON) and zearalenone (ZON) which were analyzed by HPLC methods were 210/4, 3070/88, 6100/235, and 9570/358 microg/kg in the diets fed to groups I-IV, respectively. The feeding of mycotoxin-contaminated diets did not cause gross pathological findings in the livers of the animals. Liver tissues were subjected to enzymatic, histological, and ultrastructural examinations. The percentages of the stained areas in periodic acid-Schiff (PAS), Berlin-Blue, and Masson Goldner's trichrome stainings were calculated using the AnalySIS 3.4-system. Significant histopathological findings of alterations with varying degrees in glycogen reduction and increase of hemosiderin particles were found in the liver cells of groups II, III and IV. The thickness of interlobular connective tissue septum in liver cells was significantly increased in groups III and IV. Qualitative ultrastructural alterations were observed in hepatocytes of gilts in groups III and IV. Dependent upon the mycotoxin concentration in the diet, the hepatocytes developed a dose-dependent, extensive, smooth endoplasmic reticulum, exhibited loss of ribosomes, and acquired an increased number of fatty and autophagic vacuoles. However, liver damage as measured by prominent elevated transaminase activities in serum was not detected. Together, the histopathological results provide evidence of liver dysfunction in the absence of clinical signs, especially in pigs fed higher concentrations of Fusarium toxin-contaminated wheat.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Ca(2+) Mg(2+)-ATPase; Collagen; Edible Grain; Female; Fusarium; gamma-Glutamyltransferase; Hemosiderin; Hepatocytes; Liver Diseases; Liver Glycogen; Microscopy, Electron; Swine; Swine Diseases; Trichothecenes; Triticum; Zearalenone

Results from the Bavarian Health and Food Safety Authority on contamination of cereals and cereal products from the Bavarian market with the mycotoxins deoxynivalenol (DON), zearalenone (ZEA), and ochratoxin A (OTA) and of maize meal and semolina with fumonisins (FUM) in the year 2004 are presented. Contamination rates and levels of DON, ZEA, and OTA were low and did not exceed the maximum levels. However, a 92% contamination rate and high levels of FUM in maize meal and semolina were measured. Contamination levels of mycotoxins are discussed and evaluated with respect to possible health implications for consumers.

Topics: Edible Grain; Food Contamination; Fumonisins; Fusarium; Germany; Mycotoxins; Ochratoxins; Trichothecenes; Triticum; Zea mays; Zearalenone

The Fusarium head blight (FHB)-susceptible winter wheat cv. Ritmo was inoculated with spores of Fusarium culmorum at the beginning of full blossom. Samples of whole wheat plants were taken once weekly from anthesis until harvest and subsequently fractionated into straw, glumes and spindles, which were examined for deoxynivalenol (DON) and zearalenone (ZON). Additionally, the content of crude protein (CP) and non-starch polysaccharides (NSP) was scrutinized. Synthesis of the Fusarium toxins DON and ZON generally differed in terms of date of formation and concentration. Final mean DON concentrations of 37.5, 28.1 and 5.0 mg/kg DM were measured in glumes, spindles and straw, respectively, at the time of harvest. At this time, maximal mean ZON concentrations of 587, 396 and 275 microg/kg DM in spindles, glumes and straw, respectively, were determined. Moreover, Fusarium infected wheat residues contained higher CP but lower NSP contents at the last three sampling dates. In addition, collective samples of wheat straw and chaff were taken to investigate the effect of the Fusarium contamination on their in sacco DM degradation in dairy cows. Samples were analysed for mycotoxins and selected quality parameters. The dried and milled collective samples of straw and chaff were weighed into nylon bags and subjected to ruminal incubation for 4, 8, 16, 24, 48, 72, 96 and 120 h in two dairy cows equipped with a permanent rumen cannula. Marked differences in level of mycotoxin contamination as well as in ingredient composition between the variants of straw and chaff were detected. Moreover, after 120 h rumen incubation the in sacco DM degradation of inoculated straw and chaff were lower compared to the accordant controls. The soluble fraction was increased in inoculated samples, whereas a diminishment in the potentially degradable but insoluble fraction was more pronounced. Thereby, a decrease in the potential degradability was obtained for inoculated straw and even if less pronounced for chaff compared to the non inoculated corresponding controls. In conclusion, infection with F. culmorum of wheat involves an increased risk of mycotoxin contamination in straw. Also, a Fusarium infection may have an impact on chemical composition and may result in Fusarium growth-related modifications of host cell wall components.

Topics: Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Cattle; Digestion; Female; Food Analysis; Food Contamination; Fusarium; Mycotoxins; Rumen; Solubility; Trichothecenes; Triticum; Zearalenone

Activation of the innate immune system might predispose a host to toxicant-induced inflammation. In vitro macrophage models were employed to investigate the effects of preexposure to Toll-like receptor (TLR) agonists on induction of proinflammatory cytokine gene expression by the trichothecene mycotoxin deoxynivalenol (DON) and other toxicants. Priming of the murine RAW 264.7 macrophage line or peritoneal murine macrophages with the TLR4 agonist lipopolysaccharide (LPS) at 100 ng/ml for 4, 8, and 16 h significantly increased DON-induced IL-1beta, IL-6, and TNF-alpha mRNA expression as compared to LPS or DON alone. The minimum LPS concentration for sensitization of both cell types was 1 ng/ml. LPS priming also potentiated IL-1beta mRNA induction by DON in human whole-blood cultures, suggesting the relevance of the murine findings. As observed for LPS, preexposure to TLR agonists including zymosan (TLR2), poly (I:C) (TLR3), flagellin (TLR5), R848 (TLR7/8), and ODN1826 (TLR9) sensitized RAW 267.4 cells to DON-induced proinflammatory gene expression. Amplified proinflammatory mRNA expression was similarly demonstrated in LPS-sensitized RAW 264.7 cells exposed to the microbial toxins satratoxin G, Shiga toxin, and zearalenone as well as the anthropogenic toxicants nickel chloride, triphenyltin, 2,4-dinitrochlorobenzene, and 2,3,7,8-tetrachlorodibenzodioxin. The results suggest that prior TLR activation might render macrophages highly sensitive to subsequent induction of proinflammatory gene expression by xenobiotics with diverse mechanisms of action.

Topics: Animals; Cell Line; Cytokines; Gene Expression Regulation; Humans; Lipopolysaccharides; Macrophages; Male; Mice; Mice, Inbred Strains; RNA, Messenger; Shiga Toxin; Toll-Like Receptors; Transcriptional Activation; Trichothecenes; Zearalenone

The occurrence of mycotoxins in barley, sorghum, teff (Eragrostis tef) and wheat from Ethiopia has been studied. Samples were analyzed for aflatoxin B(1) (AFB1), ochratoxin A (OTA), deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN) using high performance liquid chromatography (HPLC) and for fumonisins (FUM) using enzyme linked immunosorbent assay (ELISA). AFB1 and OTA were detected in samples of all the four crops. AFB1 was detected in 8.8% of the 352 samples analyzed at concentrations ranging from trace to 26 microg kg(-1). OTA occurred in 24.3% of 321 samples at a mean concentration of 54.1 microg kg(-1) and a maximum of 2106 microg kg(-1). DON occurred in barley, sorghum and wheat at 40-2340 microg kg(-1) with an overall incidence of 48.8% among the 84 mainly 'suspect' samples analyzed; NIV was co-analyzed with DON and was detected at 40 microg kg(-1) in a wheat sample and at 50, 380, and 490 microg kg(-1) in three sorghum samples. FUM and ZEN occurred only in sorghum samples with low frequencies at concentrations reaching 2117 and 32 microg kg(-1), respectively. The analytical results indicate higher mycotoxin contamination in sorghum, which could be related to the widespread storage of sorghum grain in underground pits leading to elevated seed moisture contents. This is the first report on the occurrence of OTA in teff.

Topics: Aflatoxin B1; Chromatography, High Pressure Liquid; Edible Grain; Ethiopia; Fumonisins; Mycotoxins; Ochratoxins; Trichothecenes; Zearalenone

This paper describes the first validated method for the determination of 39 mycotoxins in wheat and maize using a single extraction step followed by liquid chromatography with electrospray ionization triple quadrupole mass spectrometry (LC/ESI-MS/MS) without the need for any clean-up. The 39 analytes included A- and B-trichothecenes (including deoxynivalenol-3-glucoside), zearalenone and related derivatives, fumonisins, enniatins, ergot alkaloids, ochratoxins, aflatoxins and moniliformin. The large number and the chemical diversity of the analytes required the application of the positive as well as the negative ion ESI mode in two consecutive chromatographic runs of 21 min each. The solvent mixture acetonitrile/water/acetic acid 79 + 20 + 1 (v/v/v) has been determined as the best compromise for the extraction of the analytes from wheat and maize. Raw extracts were diluted 1 + 1 and were injected without any clean-up. Ion-suppression effects due to co-eluting matrix components were negligible in the case of wheat, whereas significant signal suppression for 12 analytes was observed in maize, causing purely proportional systematic errors. Method performance characteristics were determined after spiking blank samples on multiple levels in triplicate. Coefficients of variation of the overall process of <5.1% and <3.0% were obtained for wheat and maize, respectively, from linear calibration data. Limits of detection ranged from 0.03 to 220 microg/kg. Apparent recoveries (including both the recoveries of the extraction step and matrix effects) were within the range of 100 +/- 10% for approximately half of the analytes. In extreme cases the apparent recoveries dropped to about 20%, but this could be compensated for to a large extent by the application of matrix-matched standards to correct for matrix-induced signal suppression, as only a few analytes such as nivalenol and the fumonisins exhibited incomplete extraction. For deoxynivalenol and zearalenone, the trueness of the method was confirmed through the analysis of certified reference materials.

Topics: Chromatography, High Pressure Liquid; Mycotoxins; Reference Standards; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Trichothecenes; Triticum; Zea mays; Zearalenone

The aim of this study was to develop a multicomponent analytical method for the determination of deoxynivalenol (DON), ochratoxin A (OTA) and zearalenone (ZEN), nivalenol (NIV), 3-acetyl-DON (3-acDON), 15-acetyl-DON (15-acDON), zearalenol (ZOL) and citrinin (CIT) in wheat. It also aimed to survey the presence and amounts of DON, OTA and ZEN in Belgian conventionally and organically produced wheat grain and in wholemeal wheat flours. After solvent extraction, an anion-exchange column (SAX) was used to fix the acidic mycotoxins (OTA, CIT), whilst the neutral mycotoxins flowing through the SAX column were further purified by filtration on a MycoSep cartridge. OTA and CIT were then analysed by high-performance liquid chromatography (HPLC) using an isocratic flow and fluorescence detection, while the neutral mycotoxins were separated by a linear gradient and detected by double-mode (ultraviolet light fluorescence) detection. The average DON, ZEN and OTA recovery rates from spiked blank wheat flour were 92, 83 and 73% (RSDR = 12, 10 and 9%), respectively. Moreover, this method offered the respective detection limits of 50, 1.5 and 0.05 microg kg-1 and good agreement with reference methods and inter-laboratory comparison exercises. Organic and conventional wheat samples harvested in 2002 and 2003 in Belgium were analysed for DON, OTA and ZEN, while wholemeal wheat flour samples were taken from Belgian retail shops and analysed for OTA and DON. Conventional wheat tended to be more frequently contaminated with DON and ZEN than organic samples, the difference being more significant for ZEN in samples harvested in 2002. The mean OTA, DON and ZEA concentrations were 0.067, 675 and 75 microg kg-1 in conventional samples against 0.063, 285 and 19 microg kg-1 in organically produced wheat in 2002, respectively. Wheat samples collected in 2003 were less affected by DON and ZEN than the 2002 harvest. Organic wholemeal wheat flours were more frequently contaminated by OTA than conventional samples (p < 0.10). The opposite pattern was shown for DON, organic samples being more frequently contaminated than conventional flours (p < 0.10).

Topics: Belgium; Chromatography, High Pressure Liquid; Fluoroscopy; Food Analysis; Food Contamination; Mycotoxins; Ochratoxins; Reproducibility of Results; Trichothecenes; Triticum; Zearalenone

Fusarium mycotoxins occur worldwide in cereal grains and animal feeds and cause outbreaks of Fusarium mycotoxicoses in humans and animals. In this study mammalian cell cultures were used to screen the cytotoxicity of the most common Fusarium mycotoxins; deoxynivalenol (DON), zearalenone (ZEN), fumonisin B(1) (FB(1)) and moniliformin (MON). The most sensitive cell line for each Fusarium mycotoxin was determined for further toxicological investigations as an alternative to whole animal testing. Chinese hamster ovary cells (CHO-K1) were found to be the most sensitive for DON and FB(1) with IC(50) values of 0.27 and 85.5 microg/ml, respectively, after 48-h exposure. The hepatocellular carcinoma cells (HepG2) showed the highest sensitivity to MON with IC(50) values of 39.5 for 48 h and 26.8 microg/ml for 72-h exposure. Balb/c mice keratinocyte cell line (C5-O) was found to be the most sensitive to ZEN with IC(50) of 24.1 microg/ml after 72-h exposure. DON was found the most cytotoxic to the cell cultures of all the mycotoxins tested, followed by MON, ZEN, and FB(1). The results indicated that CHO-K1, C5-O, and HepG2 cells were found to be the sensitive cell lines for preliminary screening of DON, ZEN and MON contaminated feed and food extracts, respectively.

Topics: Animal Feed; Animals; Biological Assay; Cell Line; Cell Line, Tumor; CHO Cells; Cricetinae; Cricetulus; Cyclobutanes; Dose-Response Relationship, Drug; Edible Grain; Food Contamination; Fumonisins; Fusarium; Humans; Inhibitory Concentration 50; Mice; Mice, Inbred BALB C; Mycotoxins; Time Factors; Trichothecenes; Zearalenone

Three human epithelial cell lines (CaCo-2, HEp-2 and HeLa) implicated as potential targets for three Fusarium toxins were tested for the extent of survival on exposure to increasing toxin concentration and incubation periods. Cytotoxicity assay using 3(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT) was carried out with deoxynivalenol (DON), T-2 toxins and zearalenone (ZON) on CaCo-2, HEp-2 and HeLa cell lines. Of the three cell lines used, HeLa was the most sensitive, eliciting cell death after 2 days exposure at 100 ng ml(-1)with T-2 toxin. HeLa was the only cell line to exhibit cytotoxicity towards ZON showing cell death at 1000 ng ml(-1)after 2 days which increased to 4 days, showing substantial cell death at 200 ng ml(-1). HEp-2 was sensitive to DON showing cell death after 2 days (100 ng ml(-1)) with complete cell death occurring at 200 ng ml(-1) after 4 days of exposure. Substantial cytoxicity of T-2 towards HEp-2 occurred after 2 days at 1000 ng ml(-1) and complete cell death occurred with 100 ng ml(-1) at day 4. The CaCo-2 cell line was generally resistant to the mycotoxins tested between 100 and 1000 ng ml(-1). This study shows that cytotoxicity of Fusarium toxins to epithelium cell lines is concentration- and time- dependant and results from ZON-HeLa interaction indicate possible cell type-mycotoxin specificity.

Topics: Caco-2 Cells; Cell Death; Epithelial Cells; HeLa Cells; Humans; T-2 Toxin; Time Factors; Trichothecenes; Zearalenone

Fusarium species infestations of cereals crops occur worldwide. Fusarium toxins such as, deoxynivalenol (DON), zearalenone (ZEN) and fumonisin B1 (FB1) have been shown to cause diverse toxic effects in animals and also suspected of disease causation in humans. From the literature and mechanistic point of view, DON binds to the ribosomal peptidyl-transferase and inhibits protein synthesis specifically and DNA synthesis consequently. ZEN known to be genotoxic, binds to 17-beta-estradiol receptors, induces lipid peroxidation, cell death and inhibits protein and DNA synthesis. FB1 disrupts sphingolipid metabolism, induces lipid peroxidation altering the cell membrane and causing cell death. We intended to compare DON, ZEN and FB1 (1-150 microM) cytotoxic effect and the pathways leading to cell death and related to oxidative stress and macromolecules syntheses in a human intestinal cell line in order to tentatively classify them according to their respective potential toxicity. The comparison reveals that all three mycotoxins bear, at variable degree, the capability of inducing lipid peroxidation (MDA production) and could be classified above 10 microM in decreasing potency order FB1>DON>ZEN. This effect seems to be related to their common target that is the mitochondria as revealed by MTT test and seemingly not related to sphingoids accumulation concerning FB1. DON and ZEN also adversely affect lysosomes in contrast to FB1. The three mycotoxins inhibit protein synthesis with respective IC50 of 5, 8.8 and 19 microM for DON, FB1 and ZEN confirming that protein synthesis is a specific target of DON. DNA synthesis is inhibited by DON, ZEN and FB1 with respective IC50 of 1.7, 10 and 20 microM. However at higher concentrations DNA synthesis seems to be restored for FB1 and DON suggesting a promoter activity. Altogether the potency of the three mycotoxins in macromolecules inhibition is DON>ZEN>FB1 in Caco-2 cells. It appears then that FB1 acts rather through lipid peroxidation while DON affects rather DNA and protein synthesis.

Topics: Caco-2 Cells; Cell Survival; Coloring Agents; DNA Replication; Formazans; Fumonisins; Humans; Inhibitory Concentration 50; Malondialdehyde; Neutral Red; Oxidative Stress; Tetrazolium Salts; Thiobarbituric Acid Reactive Substances; Trichothecenes; Zearalenone

This study reports estimates on dietary exposure from the first French Total Diet Study (FTDS) and compares these estimates with both existing tolerable daily intakes for these toxins and the intakes calculated during previous French studies. To estimate the dietary exposure of the French population to the principal mycotoxins in the French diet (as consumed), 456 composite samples were prepared from 2280 individual samples and analysed for aflatoxins, ochratoxin A, trichothecenes, zearalenone, fumonisins and patulin. Average and high percentile intakes were calculated taking account of different eating patterns for adults, children and vegetarians. The results showed that contaminant levels observed in the foods examined 'as consumed' complied fully with current European legislation. However, particular attention needs to be paid to the exposure of specific population groups, such as children and vegans/macrobiotics, who could be exposed to certain mycotoxins in quantities that exceed the tolerable or weekly daily intake levels. This observation is particularly relevant with respect to ochratoxin A, deoxynivalenol and zearalenone. For these mycotoxins, cereals and cereal products were the main contributors to high exposure.

Topics: Adult; Aflatoxins; Child; Diet; Diet, Vegetarian; Environmental Exposure; Feeding Behavior; Food Contamination; Food Microbiology; France; Fumonisins; Humans; Mycotoxins; Ochratoxins; Patulin; Trichothecenes; Zearalenone

From 1999-2001 three different varieties of wheat [Contur (susceptible to Fusarium), Batis and Petrus (less susceptible to Fusarium)] were cultivated under organic and conventional conditions in order to determine mycotoxin burden. Soil quality, preceding crop and weather conditions were comparable in the different production systems. The wheat batches were analysed for moulds, and the contents of zearalenone (ZEN) and deoxynivalenol (DON). Feeding trials were carried out with growing pigs (n = 96; average initial live weight 22.2 +/- 1.5 kg [mean +/- SD]) to examine a possible influence on the animal performance and on mycotoxin residues. The data recorded were clinical conditions, performance, biochemical and hematological data. Residues of ZEN, alpha- and beta-zearalenol (ZEL) and of DON were determined in bile, liver and muscle after slaughtering. Conventionally cultivated wheat was more frequently contaminated with Fusarium and contained more frequently ZEN and DON in higher concentrations than the organically produced wheat. Hematological and biochemical parameters of pigs fed with organically cultivated diets were not different from those of conventionally fed pigs. Pigs fed with organically produced wheat showed a slightly higher daily weight gain, but a lower carcass yield than the conventionally fed animals. The highest residues of DON and total-ZEN (ZEN + alpha-ZEL + beta-ZEL) were found in bile. Bile samples of organically fed pigs contained lower concentrations of total-ZEN than those of conventionally fed pigs. Altogether, these data suggest that wheat from an organic farming does not have higher mycotoxin-contamination than wheat from the conventional farming system.

Topics: Agriculture; Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Bile; Blood Chemical Analysis; Food Contamination; Fusarium; Liver; Muscle, Skeletal; Mycotoxins; Random Allocation; Swine; Tissue Distribution; Trichothecenes; Triticum; Weight Gain; Zearalenone

The aim of the present study was to examine the effects of feeding Fusarium toxin-contaminated wheat to dairy cows on nutrient utilization in the rumen and on duodenal flow of deoxynivalenol (DON), zearalenone (ZON) and their metabolites. Six dairy cows fitted with a large rumen cannula and a simple T-shaped cannula at the proximal duodenum was used in two experiments. The experiments included a control period in which the uncontaminated control wheat was fed and a period in which the control wheat was replaced by the Fusarium toxin-contaminated wheat (8.05 and 7.15 mg DON/kg and 0.26 and 0.1 mg ZON/kg in Expts 1 and 2 respectively). The wheat portion of the daily ration amounted to 50% on a dry matter (DM) basis and rations were completed with hay or grass silage. Five of the six cows were non-lactating and the total daily DM-intake ranged between 4 and 12 kg. The pH-values and the concentration of volatile fatty acids in ruminal fluid were not significantly influenced by feeding the contaminated wheat. In contrast, the postprandial ammonia concentration was consistently higher when the mycotoxin-contaminated wheat was fed. Moreover, the flow of microbial protein and utilizable protein at the duodenum were reduced at the same time. The concentrations of DON and ZON and of their metabolites in freeze-dried duodenal digesta were either not detectable or negligible during the control periods whereas distinct concentrations were measured during the periods where the contaminated wheat was fed. DON was nearly completely metabolized to de-epoxy-DON and the flow at the duodenum ranged between 4% and 28% of DON-intake. The ZON metabolites alpha-zearalenol (ZOL) and beta-ZOL were recovered at the duodenum beside the parent toxin ZON. Their recovery as a percentage of ZON-intake ranged between 43% and 132%. In conclusion, feeding of Fusarium toxin-contaminated wheat altered the ruminal protein utilization. The question of whether this effect was a result of the mycotoxin being present in the rumen or of Fusarium growth-related structural (cell wall) changes of the wheat grain needs to be clarified. The low recovery of DON at the duodenum would indicate either a nearly complete degradation of the molecule in the rumen or an absorption by the mucosa of the rumen, whereas the higher ZON recovery would suggest a lower degradation of the parent toxin in the rumen and/or recovery of some bile-originating entero-hepatic cycling ZON/metabolites.

Topics: Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Bacterial Proteins; Cattle; Digestion; Dose-Response Relationship, Drug; Duodenum; Fatty Acids, Volatile; Female; Food Contamination; Fusarium; Hydrogen-Ion Concentration; Intestinal Absorption; Random Allocation; Rumen; Trichothecenes; Triticum; Zearalenone

Two feeding experiments with female weaned piglets were carried out applying a complete two by two factorial design to investigate the effects of the dietary inclusion of 500 g/kg Fusarium toxin contaminated maize (8.6 mg/kg deoxynivalenol (DON); 1.2 mg/kg zearalenone (ZON)) and of 4 g/kg aluminosilicate (AS) as a detoxifying agent. The resulting four diets were fed ad libitum to a total of 80 piglets (20 piglets per group, allotted to a total of 20 pens) covering a live weight range of 10.5 +/- 1.3 to 27.5 +/- 4.4 kg in experiment 1, and to a total of 48 piglets (12 piglets per group, allotted to 12 pens) covering a live weight range of 9.7 +/- 1.8 to 21.4 +/- 4.8 kg in experiment 2. The animals of experiment 1 were slaughtered on days 34-36 of feeding the experimental diets. The mycotoxin analyses revealed that the control maize also contained considerable concentrations of Fusarium toxins, but the differences in DON and ZON concentrations between control and contaminated diets were sufficiently high to demonstrate both dose-related toxin effects. Voluntary feed intake and live weight gain of the animals were significantly reduced by the inclusion of Fusarium toxin contaminated maize into the diets in both experiments, while a significantly decreased feed to gain ratio was found in experiment 1. Furthermore, the relative weight of the uterus, stomach and heart of the animals fed the contaminated maize containing diets were significantly increased. Serum albumin concentrations and the activity of GLDH were significantly reduced by the inclusion of the contaminated maize. The addition of AS to the Fusarium toxin contaminated diets did not prevent or alleviate any of the mentioned effects. Moreover, the feed intake tended to be decreased by this supplementation in both experiments, while a significantly decreased feed to gain ratio was indicated for this factor in experiment one as well. The serum concentration of albumin and the activities of ASAT and gammaGT were significantly increased if AS was present in the diets while serum concentration of cholesterol and alpha-tocopherol were decreased significantly or in tendency, respectively. The concentrations of retinol and retinyl esters in liver and serum were not altered by the treatments. The analysed concentrations of zearalenone (ZON) and its metabolites in the bile fluid clearly indicated the differences in dietary ZON concentrations and showed that AS was ineffective in preventing the absorption of th

Topics: Aluminum Silicates; Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Animals, Newborn; Dose-Response Relationship, Drug; Eating; Female; Food Contamination; Fusarium; Intestinal Absorption; Liver; Organ Size; Random Allocation; Swine; Treatment Outcome; Trichothecenes; Weaning; Weight Gain; Zea mays; Zearalenone

A reliable, sensitive and selective method was developed to determine different Fusarium mycotoxins (trichothecenes Type A and B, zearalenone) simultaneously in cereals and cereal-based samples using liquid chromatography with tandem mass spectrometry (LC-ESI-MS/MS). Sample preparation is based on a standard solvent extraction step followed by two different kinds of solid-phase clean-up procedures: using a multifunctional MycoSep material for trichothecenes and zearalenone. The average recoveries for trichothecenes ranged from 65% for nivalenol (NIV) up to 96% for deoxynivalenol (DON) and 89% for zearalenone (ZON). The limit of quantification varied between 0.02 and 10 ppb for each substance. In addition, a screening survey with 685 samples was carried out to compare contents of T-2 toxin and deoxynivalenol and to investigate potential coherence in contamination pattern.

Topics: Chromatography, Liquid; Edible Grain; Flour; Food, Fortified; Fusarium; Mass Spectrometry; T-2 Toxin; Trichothecenes; Triticum; Zearalenone

A total of 36 gilts (103 +/- 6 kg) were divided into four groups and fed diets with increasing proportions of a Fusarium toxin contaminated wheat over a period of 35 days. The concentrations of the indicator toxins deoxynivalenol (DON) and zearalenone (ZON) which were analyzed by HPLC methods were 210 and 4, 3070 and 88, 6100 and 235 and 9570 and 358 mug.kg(-1) diet fed to groups 1-4 respectively. Feed was partially refused during the first 21 days of the experiment by groups 2, 3 and 4 where two, three and six out of nine gilts were affected. No signs of hyperestrogenism or uterotrophic effects were observed due to dietary treatments. Blood serum, urine, bile and liver were analyzed for residues of DON, ZON and their metabolites. DON and its de-epoxidized metabolite (de-epoxy-DON) were detected in all analyzed specimens and increased in a significantly linearly related fashion. Alpha-zearalenol (alpha-ZOL) and beta-ZOL could be detected besides the parent toxin ZON, but only in bile and urine. In conclusion, the impact of dietary treatments on the performance parameters was most pronounced in the highest exposed group. The maximum ratio between DON concentration in liver and diet was 0.0013, and suggests that a possible contamination of pig liver with DON is negligible and does not contribute significantly to human DON exposure.

Topics: Animal Feed; Animals; Bile; Chromatography, High Pressure Liquid; Diet; Eating; Estrus; Female; Food Contamination; Fusarium; Liver; Mycotoxins; Swine; Trichothecenes; Triticum; Weight Gain; Zearalenone

Filamentous fungi are cosmopolitan microorganisms found in almost all environments. It should be pointed out that occurance of moulds on food or feed may cause health disorders in humans and animals. Mycoflora appears as a source of toxic methabolites, mycotoxins, which hepatotoxic, genotoxic, nefrotoxic and carcinogenic abilities were already proven in several studies. Hense mycological analysis of cereal grains raises as an important manner in evaluation of food and feed health features. Among the most frequent cereal contaminants Alternaria, Aspergillus, Fusarium and Penicillium strains are mentioned. Due to their ability to grow on cereals during both its field growth and storage, Fusarium moulds occure to be an important contamination factors in food and feed industry. In this study Fusarium strains isolates from wheat and maize were examined in order to recognize their abilities to produce two toxins: zearalenon (ZEA) and deoxynivalenole (DON). Mycological analysis shown differentiation within fungal microflora occuring in samples of different storage conditions, where Fusarium strains represented aproximately 20-70% of all mould species present. In purpose of Fusarium strains species evaluation, isolates were mycologically analysed. In the second step of the project, toxicological screening of isolates was performed using Thin Liquid Chromatography (TLC) evaluating toxigenic potential of single strains' production of ZEA and DON. This data gives the possibility of pointing the most toxigenic strains and also shows differentiations in their occurance in cereals. This paper presents introductory research data, which can be useful in recognition of cereal contamination with moulds and their toxic methabolites.

Topics: Chromatography, Thin Layer; Fusarium; Pilot Projects; Trichothecenes; Triticum; Zearalenone

Fusarium species cause not only root, stem and ear rot with severe reductions in crop yield, they produce also toxic secondary metabolites (mycotoxins) such as deoxynivalenol (DON) and zearalenone (ZEA). During several growing seasons the presence of Fusarium spp was followed up. DON and ZEA were determined and related to infection levels. The distribution of DON and ZEA in the different plant parts was studied as well as the influence of the ensiling process on the mycotoxin content. More or less important varietal differences in susceptibility for Fusarium spp. could be detected. DON and ZEA were clearly present in most of the analysed samples. No clear relationship could be detected between visual disease symptoms and mycotoxin content. The accumulation of DON and ZEA was different for the analysed aerial plant parts. The ensiling process gave no reduction of the mycotoxin content.

Topics: Belgium; Consumer Product Safety; Food Contamination; Food Preservation; Fusarium; Incidence; Mycotoxins; Trichothecenes; Zea mays; Zearalenone

The purpose of this study was to develop an LC/MS assay to accurately detect three mycotoxins produced by Fusarium graminearum in various matrices. Using different LC conditions, deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-ADON), and zearalenone (ZEN) were detected in four different matrices (fungal liquid cultures, maize grain, insect larvae and pig serum). The sensitivity of MS detection allowed us to detect concentrations as low as 8 ppb of DON and 12 ppb of ZEN. A very small quantity of matrix was therefore necessary for successful analysis of these toxins and a variety of experimental situations were successfully investigated using this technique. Production of 15-ADON and butenolide was monitored in a liquid culture of F. graminearum under controlled conditions. Using simple extraction procedures, the differential accumulation of DON and 15-ADON was followed in inoculated maize genotypes varying in susceptibility to F. graminearum. Toxicokinetic studies were carried out with maize insect pests reared continually on artificial diets containing ZEN and suggested that larvae may possess the ability to degrade ZEN. Finally, persistence of DON was assessed in pigs fed diet supplemented with DON, results indicated that DON accumulates quickly in pig blood and then levels decline progressively for 12 hours thereafter. The LC/MS study reported here is very useful and flexible for the detection of these mycotoxins in different media and at very low concentrations.

Topics: Animals; Biological Assay; Fusarium; Gas Chromatography-Mass Spectrometry; Insecta; Male; Mycotoxins; Swine; Trichothecenes; Zea mays; Zearalenone

The effect of five essential oils (oregano, cinnamon, lemongrass, clove and palmarose) on growth rate, zearalenone (ZEA) and deoxynivalenol (DON) production by Fusarium graminearum strains was assessed.. The influence of the essential oils was tested on irradiated maize at two concentrations (500 and 1000 mg kg-1), at different water activity (aw) (0.95 and 0.995) and temperature (20 and 30 degrees C) levels. At 0.995 aw all essential oils tested had an inhibitory effect on growth rate of F. graminearum at both temperatures studied. At this aw level, DON production in general was inhibited by all essential oils at 30 degrees C and, although palmarose and clove were the only essential oils with statistically significant inhibitory effect on ZEA production, an inhibitory trend was observed when cinnamon and oregano oils were added to maize grain.. Antifungal and antimycotoxigenic activity of the essential oils assayed was shown to depend on environmental conditions.. It is apparent that essential oils should be considered as alternative preharvest natural fungicides. Further investigation on natural maize grain might be useful to study the effectiveness of these essential oils in the presence of natural mycoflora of maize grain.

Topics: Antifungal Agents; Cinnamomum zeylanicum; Cymbopogon; Food Microbiology; Fusarium; Oils, Volatile; Origanum; Syzygium; Temperature; Trichothecenes; Zea mays; Zearalenone

The objective of this study was to follow the mycotoxin formation and changes in nutrient composition of wheat (cv. Ritmo) artificially inoculated with Fusarium culmorum. From anthesis until harvest, samples were taken once a week from the inoculated and control plots. The investigations were focused on monitoring the progression of the contamination of the wheat kernels with deoxynivalenol (DON) and zearalenone (ZON). Both the uncontaminated control kernels and the contaminated kernels were examined also for the presence of zearalenone-4-beta-D-glucopyranoside and several trichothecenes at harvest. Furthermore, the impact of the Fusarium inoculation on some nutrients as starch, crude protein, amino acid composition, crude ash, non starch polysaccharides (NSP) as well as viscosity and thousand seed weight (TSW) was examined. Also proteolytic and amylolytic activity as well as the NSP-degrading enzyme activities of inoculated and control samples were analysed at the time of harvest. DON was detected in higher concentrations and in earlier stages, while ZON was found later and in smaller amounts. On average 7.79 mg/kg DM of DON and 100 microg/kg DM of ZON were found in the inoculated kernels at the time of harvest. Neither in the contaminated nor in the control samples glucose conjugates of ZON (Zearalenone-4-beta-D-glucopyranoside) were detected. Moreover, the infection with Fusarium culmorum had pronounced effects on some quality parameters. The crude protein content of the inoculated kernels showed significantly higher values over the whole period compared to the control kernels. The protein content of the inoculated kernels amounted 13.9% DM at harvest, while only a concentration of 12.5% DM was detected in the control samples. Similarly, in almost all stages of development the crude ash content of inoculated samples was higher than in control samples. These distinct differences in kernel composition resulted possibly from the changes of the thousand seed weight. In the present work the grain harvested from the control plots showed a significantly higher TSW (24.2 g) as compared to their inoculated counterparts (15.5 g). Despite lower extract viscosity of inoculated samples at time of harvest, the content of soluble NSP of inoculated plots was higher than in control samples at the same time. Moreover, inoculation resulted in markedly increased activities of protease, amylase and several NSP-degrading enzyme activities. This would suggest that the cell w

Topics: Animal Feed; Food Analysis; Food Contamination; Fusarium; Nutritive Value; Rain; Seeds; Solubility; Temperature; Time Factors; Trichothecenes; Triticum; Viscosity; Zearalenone

An experiment was conducted to investigate the effects of feeding grains naturally contaminated with Fusarium mycotoxins on growth and immunological parameters of broiler chickens. Three hundred sixty, 1-d-old male broiler chicks were fed 1 of 4 diets containing grains naturally contaminated with Fusarium mycotoxins for 56 d. The diets included (1) control; (2) low level of contaminated grains (5.9 mg/kg deoxynivalenol (DON), 19.1 mg/kg fusaric acid (FA), 0.4 mg/kg zearalenone, and 0.3 mg/kg 15-acetyldeoxynivalenol; (3) high level of contaminated grains (9.5 mg/kg DON, 21.4 mg/kg FA, 0.7 mg/kg zearalenone, and 0.5 mg/kg 15-acetyldeoxynivalenol); and (4) high level of contaminated grains + 0.2% polymeric glucomannan mycotoxin adsorbent (GM polymer). Body weight gains and feed consumption of chickens fed contaminated grains decreased linearly with the inclusion of contaminated grains during the grower phase (d 21 to 42). Efficiency of feed utilization, however, was not affected by diet. Production parameters were not significantly affected by the supplementation of GM polymer to the contaminated grains. Peripheral blood monocytes decreased linearly in birds fed contaminated grains. The feeding of contaminated diets linearly reduced the B-cell count at the end of the experiment, whereas the T-cell count on d 28 responded quadratically to the contaminated diets. The feeding of contaminated diets did not significantly alter serum or bile immunoglobulin concentrations, contact hypersensitivity to dinitrochlorobenzene, or antibody response to SRBC. Supplementation with GM polymer in the contaminated diet nonspecifically increased white blood cell count and lymphocyte count, while preventing mycotoxin-induced decreases in B-cell counts. It was concluded that broiler chickens are susceptible during extended feeding of grains naturally contaminated with Fusarium mycotoxins.

Topics: Animal Feed; Animals; Chickens; Dermatitis, Contact; Energy Intake; Food Contamination; Fusarium; Immunophenotyping; Lymphocytes; Male; Meat; Mycotoxins; Organ Size; Poultry Diseases; Trichothecenes; Weight Gain; Zearalenone

1. Diets with increasing proportions of Fusarium-toxin-contaminated wheat were fed to Pekin ducks for 49 d in order to titrate the lowest effect level. Dietary deoxynivalenol (DON) and zearalenone (ZON) concentrations were successively increased up to 6 to 7 mg/kg and 0.05 to 0.06 mg/kg, respectively. 2. Feed intake, live weight gain and feed to gain ratio were not influenced by dietary treatment. 3. Gross macroscopic inspection of the upper digestive tract did not reveal any signs of irritation, inflammation or other pathological changes. The weight of the bursa of Fabricius, relative to live weight, decreased in a dose-related fashion. Activities of glutamate dehydrogenase and gamma-glutamyl-transferase in serum were either unaffected or inconsistently affected by dietary treatments. 4. Concentrations of DON and of its de-epoxydised metabolite in plasma and bile were lower than the detection limits of 6 and 16 ng/ml, respectively, of the applied high performance liquid chromatography (HPLC) method. 5. ZON or its metabolites were not detectable in plasma and livers (detection limits of the HPLC method were 1, 0.5 and 5 ng/g for ZON, alpha-zearalenol (alpha-ZOL) and beta-zearalenol (beta-ZOL), respectively). Concentrations of ZON, alpha-ZOL and beta-ZOL in bile increased linearly with dietary ZON concentration. The mean proportions of ZON, alpha-ZOL and beta-ZOL of the sum of all three metabolites were 80, 16 and 4%, respectively. 6. Taken together, it can be concluded that dietary DON and ZON concentrations up to 6 and 0.06 mg/kg, respectively, did not adversely affect performance and health of growing Pekin ducks.

Topics: Analysis of Variance; Animal Feed; Animals; Biotransformation; Bursa of Fabricius; Dose-Response Relationship, Drug; Ducks; Food Contamination; Fusarium; Mycotoxins; Trichothecenes; Triticum; Weight Gain; Zearalenone

The potential for the Fusarium mycotoxins 4-deoxynivalenol (DON) and zearalenone (ZON) to enter the human food chain through contaminated eggs was assessed using a controlled feed study. Four groups of laying hens (eight in each group) were fed a diet that included differing amounts of naturally contaminated wheat containing DON ( approximately 20 mg kg(-1)) and ZON (0.5 mg kg(-1)). Eggs were collected and pooled from each group on a daily basis. Pooled samples were analyzed by liquid chromatography with mass spectrometry detection (LC-MS/MS). The method allowed DON, other type B trichothecenes, ZON, and its metabolites to be determined in a single multi-residue analysis. The selectivity of the MS/MS procedure allowed cleanup to be minimized (for DON, cleanup by immunoaffinity column was used) or eliminated (for ZON). The limits of detection of 0.01 microg kg(-1) for DON and 0.1 microg kg(-1) for ZON in eggs were lower than previously published methods. None of the samples analyzed had detectable levels of ZON or its metabolites. Although maximum levels of DON contamination (10 mg kg(-1) feed) were relatively high, no adverse effects were observed on egg production. On the basis of the determined DON levels in the hen's diet and the determined levels of DON in the corresponding eggs, transmission rates of 15 000:1, 18 000:1, and 29 000:1 for treatment levels 5, 7.5, and 10 mg DON kg(-1) feed, respectively, were found. These results show that, although eggs could be a human exposure route for DON, the levels are insignificant compared to the other sources, although the presence of metabolites of DON was not studied.

Topics: Animal Feed; Animals; Chickens; Eggs; Female; Food Contamination; Mycotoxins; Oviposition; Trichothecenes; Triticum; Zearalenone

A survey of the occurrence of deoxynivalenol (DON) and zearalenone (ZEN) in wheat, rye, barley and maize harvested in 1989-2001 in several regions of Russia has been conducted. A total of 5652 samples of cereals were analysed for DON and ZEN by using TLC and normal-phase HPLC with UV-detector. DON was detected in 69% of 2166 samples from Krasnodar region which is considered to be the major Fusarium endemic region of Russia. The contamination levels ranged from 0.1 till 8.6 ppm, MTEL was exceeded in 37% of these samples. The positive correlation between DON concentration and a percentage of Fusaria-damaged wheat kernels has been shown. DON occurrence and contamination levels were much lower that for wheat. Based on the results of monitoring and the data of average actual consumption of wheat products in Russia, the estimated daily intake of DON per 1 kg of body weight (EDI)was calculated. EDI varied from 0.07 ug in 1990-1991 till 1.40 ug in 1992. Although average EDI were lower than adopted tolerable daily intake (TDI, 3 ug/kg body weight) EDIs for the North-Caucasian region in some cases exceeded TDI.

Topics: Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Edible Grain; Food Contamination; Humans; Russia; Trichothecenes; Zearalenone

A method for the simultaneous quantitative determination of deoxynivalenol (DON), fumonisin B1 (FB1) and zearalenone (ZEN) in maize by liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry (LC-APCIMS/MS), using stable isotopically labelled and structural analogues internal standards, is described. The procedure involves accelerated solvent extraction followed by two solid-phase clean-up steps on strong anion exchange resin and a Mycosep column. Typical recoveries were calculated by spiking blank maize at three different concentrations for deoxynivalenol (200, 400 and 1000 microg kg(-1)) at 70%, for fumonisin B1 (100, 200 and 1000 microg kg(-1)) at 90%, and for zearalenone (50, 100 and 200 microg kg(-1)) at 40%. LC-APCIMS/MS analyses were realized in collision-induced dissociation on an ion-trap instrument to provide a high degree of selectivity and sensitivity. Extraction of ions from two transition reactions, monitored by LC-APCIMS/MS for each analyte, enabled a limit of detection for DON, FB1 and ZEN at, respectively, 10, 20 and 3 microg kg(-1), and a limit of quantification at, respectively, 50, 50 and 10 microg kg(-1). The robustness of the method was also evaluated with the analysis of wheat samples.

Topics: Animals; Atmospheric Pressure; Carcinogens, Environmental; Chromatography, Liquid; Food Contamination; Fumonisins; Fusarium; Humans; Mass Spectrometry; Mycotoxins; Reproducibility of Results; Solvents; Trichothecenes; Zea mays; Zearalenone

A simple in vitro system was developed to study the efficacy of commercially available mycotoxin detoxifying agents and adsorbing substances as feed additives to detoxify deoxynivalenol (DON) and zearalenone (ZON) in situ. The in vitro model simulates the conditions (pH, temperature and transit time) of the porcine gastrointestinal tract, as pigs react most sensitively to these mycotoxins. The commercially available products were not effective in detoxifying DON and ZON under the applied conditions, while activated carbon was able to bind both toxins and cholestyramine, and a modified aluminosilicate showed good adsorption abilities for ZON. Data obtained in dose dependency studies showed an estimated adsorption capacity of cholestyramine and the modified aluminosilicate of 11.7 and 5.7 g ZON/kg detoxifying agent. The in vitro system deployed in the present study was demonstrated to be a simple, helpful tool in screening substances for their ability to detoxify DON and ZON under the simulated conditions of the porcine gastrointestinal tract. Nonetheless in vivo experiments are indispensable to proof the efficacy.

Topics: Adsorption; Aluminum Silicates; Animal Feed; Animals; Anion Exchange Resins; Buffers; Charcoal; Cholestyramine Resin; Dose-Response Relationship, Drug; Estrogens, Non-Steroidal; Food Additives; Food Contamination; Hydrogen-Ion Concentration; In Vitro Techniques; Models, Biological; Mycotoxins; Swine; Time Factors; Treatment Outcome; Trichothecenes; Zearalenone

The effects of feeding diets containing 0.01, 0.06, 0.15, 0.22 and 0.42 mg zearalenone and 0.2, 0.8, 1.0, 1.9 and 3.9 mg deoxynivalenol per kg, originating from Fusarium toxin contaminated maize, on the uterus of 50 prepubertal piglets (10 pigs per treatment; BW 32.6+/-5.4 kg; approximately 70 days of age) were investigated. The mean weight of the uteri of animals receiving the most highly contaminated diet was significantly increased at the time of slaughtering. The histological investigation showed no marked differences between the feeding groups. Histometrical parameters of the surface epithelium of the uterus, of the uterine glands and the vaginal epithelium were not altered by the treatment.

Topics: Animal Feed; Animals; Animals, Newborn; Dose-Response Relationship, Drug; Estrogens, Non-Steroidal; Female; Food Contamination; Fusarium; Image Processing, Computer-Assisted; Organ Size; Random Allocation; Swine; Trichothecenes; Uterus; Vagina; Weight Gain; Zea mays; Zearalenone

The concentrations of the Fusarium toxins deoxynivalenol (DON) and zearalenone (ZON) of a heavily contaminated wheat grain batch were followed over a period of 1 year by taking samples 15 times every 28 days. The air temperature and relative humidity at the top of the wheat batch ranged between 7 and 22 degrees C and 44 and 55%, respectively, and corresponded to a variation in the moisture content of the wheat grain between 11.5 and 12.3%. None of these fluctuations were related to ZON and DON concentrations, which varied between 0.46 and 0.66 and 15.0 and 19.5 mg/kg DM. Therefore, the data were used to analyse the error sources for the analytical results. It was found that the variance proportions due to sampling and sample preparation plus analysis were not similar for DON and ZON. The variance proportion due to sampling was found to be 0.62 for ZON, which corresponded to a variance proportion of 0.38 due to sample preparation plus analysis. In contrast, the latter variance proportion for DON was estimated to be 1.0 and consequently completely superimposed the sampling error. It is concluded that long-term storage of contaminated wheat grain does not affect the concentrations of DON and ZON considering the measured fluctuations in ambient temperature, relative humidity and moisture content of the grain. Therefore, no degradation of DON and ZON occurred during the storage of wheat for a period of one year under ambient conditions. The effects of sampling and sample preparation plus analysis on the final analytical results are different for DON and ZON and require further consideration.

Topics: Animal Feed; Animals; Food Contamination; Food Handling; Food Preservation; Fusarium; Humidity; Mycotoxins; Selection Bias; Temperature; Time Factors; Trichothecenes; Triticum; Zearalenone

Fusarium graminearum strains are well known for their role as plant pathogens and for their production of mycotoxins, and less known for their secretion of galactose oxidase, a well-studied and useful enzyme. Three galactose oxidase-producing isolates of F. graminearum were grown on rice to identify the production of zearalenone and trichothecenes through the use of thin layer chromatography and gas chromatography coupled to mass fragmentation. Detection and identification of deoxynivalenol, 3-acetyldeoxynivalenol, and zearalenone were accomplished.

Topics: Fusarium; Galactose Oxidase; Mycotoxins; Trichothecenes; Zearalenone

Selective polymeric phases intended for future use in separation/extraction of deoxynivalenol and zearalenone from beverages have been prepared. Using crystalline deoxynivalenol, zearalenone and quercetin, molecularly imprinted polymers were obtained by a non-covalent imprinting approach via a photo-initiated addition polymerization. Prepared polymers were based on 4-vinylpyridine, methacrylic acid or 2-trifluoromethylacrylic acid as the functional monomer and on ethyleneglycol dimethacrylate, trimethyltrimethacrylate or divinylbenzene as the cross-linking monomer. Selectivity of the generated molecularly imprinted polymers has been investigated by application of the prepared molecularly imprinted polymers as stationary phases in high-pressure liquid chromatography experiments. The retention and elution behaviours of the template compounds and structurally related substances were determined and compared. The results promise future application of molecularly imprinted polymers as alternative selective matrices for clean-up and enrichment of deoxynivalenol and zearalenone.

Topics: Beer; Chromatography, High Pressure Liquid; Food Contamination; Mycotoxins; Polymers; Trichothecenes; Wine; Zearalenone

Information on the contamination of Danish cereals and cereal products with Fusarium toxins is limited and the last survey is from 1984/1985. In the present study, the occurrence of deoxynivalenol (DON), nivalenol (NIV), HT-2 toxin, T-2 toxin and zearalenone (ZON) was investigated in flour of common wheat, durum wheat and rye. The samples were collected from 1998 to 2001 from both mills and the retail market in Denmark. A total of 190 flour samples were analysed for DON and NIV and about 60 samples for HT-2, T-2 toxin and ZON. DON was most frequently detected with an incidence rate of 78% over all samples for all years. The contamination level varied considerably from year to year, and for wheat and rye the highest incidence and DON concentrations were found in samples from the 1998 harvest. There were regular and heavy rainfalls in Denmark during the flowering period of the crops that year, and DON was found in all samples, with mean concentrations in wheat and rye flour of 191 microg kg(-1) (n=14) and 99 microg kg(-1) (n=16), respectively. Comparison of data from each harvest year showed higher contents of DON in samples of wheat (range 20-527 microg kg(-1)) than in rye (20-257 microg kg(-1)). Contents of NIV, HT-2 toxin and ZON in samples of wheat and rye were generally low, and even in positive samples the contents were close to the detection limit of the methods. The T-2 toxin was detected in only a few of the wheat samples and in low amounts. However, the toxin was found in about 50% of the rye samples collected during 1998-2000, with a mean content of 49 microg kg(-1) (n=25). Durum wheat flour showed the highest DON contamination level, and all samples (n=33) collected during 2000 and 2001 contained DON with means and medians above 1100 microg kg(-1). Over 70% of the samples contained more than 500 microg kg(-1) DON, and the highest observed concentration was 2591 microg kg(-1). The concentration of T-2 toxin in durum wheat flour was also high with five of the 10 analysed samples containing more than 100 g kg(-1).

Topics: Denmark; Flour; Food Contamination; Fusarium; Humans; Mycotoxins; Secale; Sweden; T-2 Toxin; Trichothecenes; Triticum; Zearalenone

A dose response study was carried out with piglets to examine the effects of increasing amounts of Fusarium toxins in the diet on performance, clinical serum characteristics, organ weights and residues of zearalenone (ZON) and deoxynivalenol (DON) and their metabolites in body fluids and tissues. For this purpose, Fusarium toxin contaminated maize (1.2 mg ZON and 8.6 mg DON per kg maize) was incorporated into a maize based diet for piglets at 0, 6, 12.5, 25 and 50% at the expense of control maize. The experimental diets were tested on 100 female piglets allotted to 20 boxes (five animals per box) covering a body weight range of 12.4 +/- 2.2 kg to 32.5 +/- 5.6 kg. Voluntary feed intake and, consequently, body weight gain of the animals receiving the highest proportion of Fusarium toxin contaminated maize were significantly decreased while the feed conversion ratio was not affected by the treatment. The mean weight of the uterus related to the body weight of the animals of the same group was increased by almost 100% as compared to the control. For this group, significantly decreased values of total serum protein were determined, while the serum activity of the liver enzyme glutamate dehydrogenase and the serum concentration of the follicle stimulating hormone were decreased for all treatment groups receiving 6% contaminated maize or more in the diet. Serum concentrations of immuneglobulins were not consistently altered by the treatment. Corresponding to the dietary exposure, increasing concentrations of ZON and alpha-zearalenol were detected in the bile fluid, liver and in pooled urine samples. The metabolite beta-zearalenol was detected only in bile fluid. The total concentration of ZON plus its metabolites in bile fluid correlated well with the diet contamination (r = 0.844). DON was found in serum, bile fluid and pooled urine samples while de-epoxy-DON was detected only in urine. The serum concentration of DON correlated well with the respective toxin intake 3-4 h prior to slaughtering (r = 0.957). For all mentioned analyses of residues it has to be noted that toxin residues were detectable even if negligible concentrations were present in the diet.

Topics: Animal Feed; Animals; Animals, Newborn; Dose-Response Relationship, Drug; Drug Residues; Eating; Female; Food Contamination; Fusarium; Liver; Mycotoxins; Organ Size; Random Allocation; Swine; Trichothecenes; Weaning; Weight Gain; Zea mays; Zearalenone

The influence of heating temperature and time on deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEA) contents in naturally co-contaminated barley and wheat was investigated intending to establish the basis for a decontamination model of Fusarium mycotoxins in cereals. The standard toxins and whole barley powder samples were heated in a convection oven at 140, 160, 180, 200, or 220 degrees C, and kernel subsamples (200 g each) were roasted in an experimental rotary gas-fired roaster at 150, 180 or 220 degrees C. All treatments resulted in a time-temperature-dependent decomposition of the toxins; the logarithm of the toxin remaining % presented a linear relationship with heating time. The lines equations were used to estimate the half (H) and decimal (D) decomposition times (time required to destroy 50 or 90% of the toxin, respectively). DON and NIV H and D decomposition times were similar and 50% shorter for heated standards than for whole barley powder. ZEA standard values were considerably longer, while whole barley powder values were comparable with those of DON and NIV. At 220 degrees C, D decomposition times of DON, NIV and ZEA heated standards were 11, 10 and 85 min, respectively, while the values obtained in whole barley powder were the same for the three toxins (25 min). The determination of H and D decomposition values constitutes a basis to understand the heating stability nature of each toxin.

Topics: Decontamination; Drug Stability; Food Contamination; Food Handling; Fusarium; Heating; Hordeum; Humans; Mycotoxins; Temperature; Trichothecenes; Triticum; Zearalenone

Three hundred sixty, 1-d-old male broiler chicks were fed diets containing grains naturally contaminated with Fusarium mycotoxins for 56 d. The four diets included control (0.14 mg/kg deoxynivalenol, 18 mg/ kg fusaric acid, < 0.1 mg/kg zearalenone), low level of contaminated grains (4.7 mg/kg deoxynivalenol, 20.6 mg/kg fusaric acid, 0.2 mg/kg zearalenone), and high level of contaminated grains without (8.2 mg/kg deoxynivalenol, 20.3 mg/kg fusaric acid, 0.56 mg/kg zearalenone) and with (9.7 mg/kg deoxynivalenol, 21.6 mg/kg fusaric acid, 0.8 mg/kg zearalenone) 0.2% esterified-glucomannan polymer derived from Saccharomyces cerevisiae1026 (E-GM). Body weight gain and feed consumption responded in a significant quadratic fashion to the inclusion of contaminated grains during the finisher period. Efficiency of feed utilization, however, was not affected by diets. The feeding of contaminated grains in the finisher period also caused significant linear increases in blood erythrocyte count and serum uric acid concentration and a significant linear decline in the serum lipase activity. Dietary inclusion of contaminated grains resulted in a significant quadratic effect on serum albumin and y-glutamyltransferase activity. Blood hemoglobin and biliary IgA concentrations, however, responded in significant linear and quadratic fashions. Supplementation of E-GM counteracted most of the blood parameter alterations caused by the Fusarium mycotoxin-contaminated grains and reduced breast muscle redness. It was concluded that broiler chickens may be susceptible to Fusarium mycotoxicoses when naturally contaminated grains are fed containing a combination of mycotoxins.

Topics: Animal Feed; Animals; Chickens; Color; Eating; Erythrocyte Count; Erythrocyte Indices; Food Contamination; Fusaric Acid; Fusarium; gamma-Glutamyltransferase; Hematocrit; Hemoglobins; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Male; Muscle, Skeletal; Mycotoxins; Trichothecenes; Zearalenone

16-wk experiment with laying hens was carried out to examine the effects of feeding of mycotoxin-contaminated maize (CM) on performance, nutrient digestibility, weight of organs, serum chemical parameters, and antibody titers to Newcastle disease virus (NDV) in serum. Also tested were fimbrien antigen K88 in egg yolk and zearalenone (ZON) residues in eggs and tissues. The Fusarium-toxin-contaminated maize contained 17,630 microg deoxynivalenol and 1,580 microg ZON/kg. Moreover, Mycofix Plus (MP), a so-called detoxifying agent, was added to both the uncontaminated control (UCM) and to the CM diet (70% dietary maize inclusion). Each of the four resulting diets (UCM, UCM-MP, CM, CM-MP) was tested on 25 laying hybrids (Lohmann Brown). Feeding of the CM diets significantly depressed feed intake compared to the control groups by approximately 5%. This was mainly due to the effects observed at the beginning of the experiment. Daily egg mass production/hen was 56.6, 58.4, 53.9, and 55.2 g in groups UCM, UCM-MP, CM and CM-MP, respectively. Nutrient digestibility and metabolizability of gross energy were slightly depressed by feeding the CM diets and improved by MP addition. Feeding of the CM diets resulted in a significant decrease in serum titers to NDV and to an increase in yolk titers to antigen K88. No residues of ZON or of its metabolites were found in yolk, albumen, abdominal fat, breast meat, follicles greater than 1 cm in diameter, ovaries including follicles smaller than 1 cm in diameter, magnum, and serum. ZON and alpha-zearalenol (alpha-ZOL) were detected in livers of hens fed the CM diets at mean concentrations of 2.1 and 3.7 microg/kg, respectively. It was concluded that feeding maize which was highly contaminated with Fusarium mycotoxins adversely influenced performance of hens and modulated immune response. At the given level of zearalenone and at the indicated detection limits, no residues of ZON and its metabolites were found in eggs. The effects of the tested detoxifying agent were quite mycotoxin-independent.

Topics: Animal Feed; Animal Nutritional Physiological Phenomena; Animals; Antibodies; Antigens, Bacterial; Chickens; Digestion; Drug Residues; Eating; Eggs; Escherichia coli Proteins; Estrogens, Non-Steroidal; Female; Fimbriae Proteins; Food Contamination; Fusarium; Iodophors; Liver; Mycotoxins; Newcastle disease virus; Organ Size; Oviposition; Tissue Distribution; Trichothecenes; Zea mays; Zearalenone

A total of 60 samples of wheat flour were collected during the first 6 months of 1999 from mills and food stores in an area in southwest Germany. Samples included whole-grain and two types of white flour with these three groups characterized by a high, medium and low ash content. The contents of deoxynivalenol (DON), nivalenol (NIV), 3- and 15-acetyldeoxynivalenol, HT-2 toxin (HT-2), T-2 toxin (T-2) and fusarenon-X (FUS-X) were determined by gas chromatography/mass spectrometry, and those of zearalenone (ZEA), alpha- and beta-zearalenol (alpha- and beta-ZOL) by high performance liquid chromatography with fluorescence detection. FUS-X, alpha- and beta-ZOL were not detected in any sample. Based on incidence and level, DON was the predominant toxin followed by NIV and ZEA for all three flour types. The overall degree of toxin contamination was lower with decreasing ash content. This suggests a localization of the toxins analyzed primarily in the outer parts of the original wheat kernels. The median DON content was significantly (P<0.05) higher for wheat flour originating from wheat of conventional than of organic production.

Topics: Chromatography, High Pressure Liquid; Flour; Food Analysis; Food Microbiology; Fusarium; Gas Chromatography-Mass Spectrometry; Germany; Mycotoxins; T-2 Toxin; Trichothecenes; Triticum; Zearalenone; Zeranol

3970 feed samples were analysed for the fusariumtoxins deoxynivalenol and zearalenone. 979 (24.6%) of the submitted feed samples from farms with case history (swine herds: decreased feed intake and daily weight gain, vomiting, cannibalism, impaired fertility, small litter size, weakness of newborn piglets, prolonged oestrus of sows; cattle: persistence of follicular ovarian cysts, enteritis, decreased slaughter weight, feed refusal). 74% of the samples contained cereal and corn ingredients respectively or were complete diet samples from swine herds. Based on economic losses the fusariumtoxin deoxynivalenol is most important in Austrian husbandry and particularly found in maize, cornsilage, wheat and oat.

Topics: Animal Feed; Animals; Austria; Body Weight; Edible Grain; Female; Fusarium; Pregnancy; Swine; Swine Diseases; Trichothecenes; Zea mays; Zearalenone

The galactose oxidase-producing fungus Dactylium dendroides was re-identified as a Fusarium species. Fungi of this genus are well known for the production of mycotoxins. Verification of growth of this fungus on rice, corn and liquid medium described for the production of galactose oxidase is provided to determine whether the fungus could produce Fusarium toxins, namely, moniliformin, fusaric acid, fumonisin, zearalenone and the trichothecenes, deoxynivalenol, 3-acetyldeoxynivalenol, fusarenone, nivalenol, diacetoxyscirpenol, neosolaniol, and toxin T-2. Under the culture conditions used, deoxynivalenol, 3-acetyldeoxynivalenol and zearalenone were detected in the fungal culture medium. The finding is consistent with the hypothesis that the fungus is in fact a Fusarium species.

Topics: Animals; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Fusarium; Galactose Oxidase; Gas Chromatography-Mass Spectrometry; Male; Mycotoxins; Rats; Rats, Wistar; Skin; Skin Irritancy Tests; Trichothecenes; Zearalenone

Twenty-eight laboratories from 12 different countries participated in an interlaboratory study for the determination of the Fusarium mycotoxin zearalenone (ZON) in maize and deoxynivalenol (DON) in maize and wheat employing their usual in-house methods. The aim of this study was to obtain information about the state-of-the-art of ZON and DON analysis in cereals and to support a knowledge and experience exchange between the participating laboratories in the field of mycotoxin analysis. Eight different sample types were distributed to the participants, 'blank' materials, spiked samples (102 microg/kg ZON in maize and 475 microg/kg DON in wheat) and naturally-contaminated maize and wheat. For the final separation and quantification either gas chromatography (GC), high performance liquid chromatography (HPLC), thin layer chromatography (TLC) or enzyme linked immunosorbent assays (ELISA) were employed by the participating laboratories. Coefficients of variation (CV) between laboratory mean results (outliers rejected) ranged from 28 to 41% for ZON and from 32 to 38% for DON. The results are close to the between laboratory CV criteria of 40% for DON and ZON at concentration levels of >100 microg/kg established by the CEN in 1999. A good trueness was obtained for the wheat samples spiked at 475 microg/kg DON. However, a significant deviation at p = 0.01 from the respective target value was observed for the maize samples spiked at 102 microg/kg ZON. The high CVs can be traced back to problems occurring by determination of the concentration of the participants' own calibrant solutions. Additionally, the variability of the results is strongly influenced by the use of different final separation and quantification procedures.

Topics: Calibration; Estrogens, Non-Steroidal; Food Analysis; Food Contamination; Humans; Laboratories; Quality Assurance, Health Care; Trichothecenes; Triticum; Zea mays; Zearalenone

The occurrences and concentrations of trichothecenes, ochratoxin A and zearalenone in Finnish cereal samples are presented in this study. Furthermore, injections by moulds, especially Fusarium contamination of grains in the same samples, are reported. In total 68 cereal samples, including 43 rye, 4 wheat, 15 barley and 6 oats samples, were collected after a cool and very rainy growing season in 1998. A gas chromatograph combined with a mass spectrometric detector was used for determination of seven different trichothecenes. A high performance liquid chromatograph with a fluorescence detector was used for ochratoxin A and zearalenone determination. For the identification of moulds, the grain samples were incubated and the moulds were isolated and identified by microscopy. The analytical methods were validated for mycotoxin analysis and they were found to be adequately reliable and sensitive. Heavy rainfalls in the summer and autumn of 1998 caused abundant Fusarium mould infection in Finnish cereals, particularly in rye. Fusarium avenaceum was the most common Fusarium species found in cereals. However, the mycotoxin concentrations were very low and only deoxynivalenol, nivalenol and HT-2 toxin were detected. Deoxynivalenol was detected in 54 samples in the concentration range 5-111 microg/kg. Nivalenol and HT-2 toxin were detected in three and two samples, respectively, in the concentration range 10-20 microg/kg.

Topics: Chromatography, Gas; Chromatography, High Pressure Liquid; Edible Grain; Food Microbiology; Fusarium; Humans; Mass Spectrometry; Ochratoxins; Rain; Reproducibility of Results; Sensitivity and Specificity; T-2 Toxin; Trichothecenes; Zearalenone

The effects of moisture, pH and heat on the stability of nivalenol (NIV), deoxtnivalenol (DON) and zearalenone (ZEN) present as natural contaminants of ground maize were measured for different periods. Standard solution tests were also performed to measure pH, salt and temperature effects on NIV and DON. The solution tests showed NIV and DON to be relatively stable in buffer solutions over the pH range 1-10. Quite harsh conditions (pH 12, high salt concentration, 80 degrees C, prolonged exposure) were needed to give substantial breakdown. In the ground maize substrate, these toxins were further stabilized relative to the solution tests. NIV and DON were both reduced (range 60-100%) by treatment with aqueous bicarbonate solution at 10, 20 or 50% of the ground maize dry weight, and subsequent heating at 80 or 110 degrees C for 2 and 12 days. There was no measurable reduction at lower test temperatures (20, 40 degrees C). NIV (but not DON) also showed some reduction following addition of water and heating at 80 or 110 degrees C for 12 days. ZEN content was not reduced even by 12 days of heating at 110 degrees C after treatment with a sodium bicarbonate solution.

Topics: Analysis of Variance; Bicarbonates; Chromatography, High Pressure Liquid; Fusarium; Hot Temperature; Humans; Hydrogen-Ion Concentration; Mycotoxins; Sodium Chloride; Time Factors; Trichothecenes; Water; Zea mays; Zearalenone

Based on a feeding trial using 27 lactating "Simmental-cows" the effect of naturally contaminated feed with deoxynivalenol (DON) as well as zearalenone (ZON) regarding production parameters was examined. 3 groups of cows according to lactation number, milk yield (kg ECM) and body mass were used. The average daily intake of DON in group K was 12.4 mg, in group T 14.1 mg and in group M 14.3 mg and ZON in group K was 12.4 mg, in group T 0.67 mg and in group M 0.68 mg respectively. The feed of animals of group M was supplemented with "Mycofix Plus" as mycotoxin inactivator. The red and white blood picture including the thrombocytes were in all groups within the normal range. Concerning enzymes (GGT, AP) and metabolites (GLUC, TBIL, UREA, CREA) the mean values of the 3 groups were in the normal range. Slightly increased were the mean values of all groups in respect to the AST- and GLDH-activities. Volatile fatty acids of the rumen content were significantly highest in group M, also the number of dead rumen infusoria was significantly decreased, but the counts of small sized infusoria increased. The study has shown that "Mycofix Plus" might be able to enhance the activity of rumen flora concerning detoxification of mycotoxins in feed of dairy cows.

Topics: Animals; Body Weight; Cattle; Dairying; Erythrocyte Count; Female; Food Microbiology; Lactation; Leukocyte Count; Milk; Mycotoxins; Platelet Count; Rumen; Trichothecenes; Zearalenone

The interactive effect of combinations of the Fusarium mycotoxins deoxynivalenol (DON), zearalenone (ZEA) and fumonisin B1 (FB1) on growth of brewing yeasts was examined. Yeast growth was assessed by measurement of dry weight or relative growth, cell number, viability and conductance change of the growth medium using direct and indirect methods. The interactive effect of a combination of these mycotoxins was subject to the ratio of toxins in the mixture and the toxicity of individual toxins on yeast growth. When a combination of mycotoxins at low concentration was added into the growth medium, no significant inhibitory effect on growth was observed compared to controls. However, when a combination of high concentrations of DON and ZEA which individually inhibited yeast growth was examined, the interactive effect was shown to pass from antagonism to synergism depending on the ratio of the toxins in the mixture. As a synergistic interaction between these Fusarium mycotoxins was observed only at high concentrations, which were far higher than would be expected in good quality grain, they are not a concern when related to yeast growth under the brewing conditions studied.

Topics: Beer; Carboxylic Acids; Colony Count, Microbial; Drug Synergism; Fumonisins; Fusarium; Mycotoxins; Trichothecenes; Yeasts; Zearalenone

One hundred and ninety seven maize samples representing different cultivars, collected from different agroclimatic regions of Karnataka (India) were analysed for moisture content, mould incidence, ergosterol and extent of mycotoxin contamination. Moisture content determination by the hot-air oven method revealed significantly high levels of moisture content (15-18%) in 34 (17%) samples, which exceeded the permissible limit for safe storage. Ergosterol quantification by HPLC revealed the presence of ergosterol in many samples collected from rural areas of Karnataka irrespective of the moisture content. Mould enumeration based on blotter and agar plating methods revealed the association of 24 diverse species of both field and storage moulds belonging to 14 genera. Mycotoxins analyses using monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) and thin-layer chromatography (TLC) revealed mycotoxin contamination in 69 (34.8%) samples. Maize samples with a high incidence of diverse species of moulds and alarmingly high levels of mycotoxins in many samples indicate the need for proper surveillance and monitoring exclusively for the prevention of moulds and mycotoxins in maize produce in Karnataka before it reaches the consumer.

Topics: Aflatoxin B1; Citrinin; Environmental Monitoring; Ergosterol; Fungi; India; Mycotoxins; Ochratoxins; Seeds; T-2 Toxin; Trichothecenes; Water; Zea mays; Zearalenone

The effects of three regimens of cycling incubation temperatures and incubation at constant 25 degrees C on the growth of Fusarium graminearum NRRL 5883 and production of deoxynivalenol (DON) and zearalenone (ZEN) on rice were compared. The effects of low-temperature stress were also studied by incubating rice cultures at a constant 15 degrees C for 4 weeks following incubation at constant 25 degrees C for 2 weeks. Both incubation temperature and time significantly (P < or = 0.05) affected growth of F. graminearum NRRL 5883 and production of DON and ZEN. The highest amount of free ergosterol (640 microg/g culture material) that was used as a measure of fungal growth was found in cultures incubated at temperatures cycling between 15 and 30 degrees C during a 6-week period. The highest amounts of DON (1,679 microg/g culture material) and ZEN (603 microg/g culture material) were produced in cultures incubated at a constant 25 degrees C for 2 weeks prior to incubation at a constant 15 degrees C for an additional 4 weeks. Under cycling incubation temperatures, maximum amounts of DON (850 microg/g culture material) and ZEN (98 microg/g culture material) were produced in cultures incubated at temperatures cycling between 15 and 30 degrees C for 6 weeks. Overall, there was no correlation between mold growth and production of either DON or ZEN. However, DON production and ZEN production were correlated.

Topics: Chromatography, High Pressure Liquid; Culture Media; Fusarium; Oryza; Temperature; Trichothecenes; Zearalenone

The presence of mycotoxins in corn-based foods available in Argentina was determined in order to make a preliminary exposure assessment. Thirty-eight samples [corn meal ('polenta') and corn flakes] of different local brands were analysed for zearalenone, deoxynivalenol and aflatoxins by TLC and fumonisins (FB1, FB2 and FB3) by HPLC. None of the 38 samples contained any detectable amount of aflatoxins (< 2 micrograms/kg), zearalenone (< 50 micrograms/kg) and deoxynivalenol (< 50 micrograms/kg). By contrast fumonisin contamination was found in 95% of the samples. The highest fumonisin levels were found in corn meal: FB1 (range positives: 60-2860 micrograms/kg; mean positive value: 556 micrograms/kg), FB2 (61-1090 micrograms/kg; 232 micrograms/kg) and FB3 (18-1015 micrograms/kg; 150 micrograms/kg). Low levels of fumonisin B1 were detected in 16/17 corn flakes samples (2-38 micrograms/kg). Total fumonisin levels in corn meal were more than 1000 micrograms/kg in 24% (5/21) of the samples. Although it is not the staple food in Argentina, maize consumption is very important, especially among children. A daily fumonisin intake of 11.3 micrograms/kg of body weight was estimated for child consumers (1-5 years old) based on an average consumption of 200 g of corn meal/day. Calculated at an average rate for all children (consumers or not) the intake estimate was 0.9 microgram/kg of body weight.

Topics: Aflatoxins; Argentina; Carboxylic Acids; Carcinogens, Environmental; Food Contamination; Food Handling; Fumonisins; Humans; Mycotoxins; Trichothecenes; Zea mays; Zearalenone

Corn samples collected from the main production area in Argentina in 1995 were surveyed for the natural occurrence of Fusarium mycotoxins and aflatoxins. Fumonisins B1, B2 and B3 and zearalenone were found in all samples. A positive relationship was found between fumonisins B1, B2 and B3, B1 and B3, and B2 and B3. Deoxynivalenol and aflatoxins were not detected. Mycological survey has also revealed the predominance of Fusarium moniliforme. This is the first report on the simultaneous occurrence of fumonisins and zearalenone in corn from the main production area in Argentina.

Topics: Aflatoxins; Argentina; Fusarium; Humans; Mycotoxins; Trichothecenes; Zea mays; Zearalenone

A total of 237 commercially available samples of cereal-based foods including bread and related products, noodles, breakfast cereals, baby and infant foods, rice and other foods were randomly collected in southwest Germany during the first six months of 1998. The trichothecenes deoxynivalenol (DON), 3- and 15-acetyl-deoxynivalenol (3-,15-ADON), nivalenol (NIV), fusarenon-X (FUS-X), T-2 toxin (T-2) and HT-2 toxin (HT-2) were determined by gas chromatography/mass spectrometry following clean-up by a two stage solid-phase extraction. Detection limits ranged between 2 and 12 micrograms/kg. Based on all samples, the incidence of DON, HT-2, T-2, 3-ADON, 15-ADON, and NIV was at 71, 18, 4, 4, 4 and 2%, respectively; the average contents in positive samples were at 103, 16, 14, 17, 24 and 109 micrograms/kg, respectively. Fus-X was not detected in any sample. A lower (P < 0.05) DON content was found in baby and infant foods as well as in cookies and cakes compared to bread. Overall, based on the incidence and level of all six toxins, the degree of contamination was lowest in baby and infant foods. Foods produced from either white or whole grain flour did not differ (P > 0.05) with regard to the incidence and level of DON. In foods produced from cereals of organic production both the incidence and median content of DON was lower compared to conventional production. Zearalenone, alpha- and beta-zearalenol were determined by high performance liquid chromatography in 20 selected samples, mostly baby and infant foods. These toxins were not present in excess of the detection limit in any sample.

Topics: Bread; Chromatography, Affinity; Chromatography, High Pressure Liquid; Edible Grain; Food Microbiology; Fusarium; Gas Chromatography-Mass Spectrometry; Germany; Humans; Infant Food; Mycoses; Mycotoxins; Oryza; Secale; Spectrometry, Fluorescence; T-2 Toxin; Trichothecenes; Triticum; Zea mays; Zearalenone; Zeranol

The efficiency of modern sample preparation techniques are discussed and compared to well-established techniques with respect to the determination of zearalenone in corn and B-trichothecenes in wheat in the microgram/kg range. This includes the use of immuno-affinity columns and of multifunctional Mycosep columns as well as the employment of supercritical fluid extraction for the trace analysis of these major Fusarium mycotoxins. In addition, the performance of new analytical methods was investigated in an interlaboratory comparison study only recently organized by our laboratory. From both the validation data, and from the results of the intercomparison study, the suitability and competitiveness of the described methods could be clearly demonstrated.

Topics: Chromatography, Affinity; Chromatography, Gas; Chromatography, High Pressure Liquid; Edible Grain; Fusarium; Immunochemistry; Mycotoxins; Trichothecenes; Zearalenone

Thin layer chromatography (TLC) methods for identifying and quantifying deoxynivalenol (DON), fumonisin B1 (FB1) and zearalenone in grain samples were compared to immunoassay (ELISA) and high performance liquid chromatography (HPLC) methods to determine the reliability of the less expensive TLC. There was a very good agreement between levels of DON measured by TLC and competitive-direct ELISA, and between levels of fumonisin B1 measured by TLC and HPLC, over a wide range of concentrations. Correlation coefficients (Pearson's) were 0.978, 0.914 and 0.953 for DON in maize, DON in wheat and FB1 in maize respectively. A lower correlation coefficient (r = 0.672) was obtained when zearalenone was quantified by TLC and HPLC. Possible reasons for this are discussed. A cost comparison of the various methods revealed that TLC was the least expensive for sample analysis. It is recommended that researchers choose which analytical method to use based upon individual considerations of cost and precision.

Topics: Carboxylic Acids; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Enzyme-Linked Immunosorbent Assay; Fumonisins; Fusarium; Mycotoxins; Trichothecenes; Triticum; Zea mays; Zearalenone

In order to survey the natural occurrence of Fusarium mycotoxins including deoxynivalenol (DON), nivalenol (NIV), and zearalenone (ZEN) in beers consumed in Korea, a total of 54 extracts of Korean and imported beers were derivatized with heptafluorobutyric anhydride and analysed by gas chromatography-mass spectrometry with selected ion monitoring mode. DON was detected in 14 samples (26%) and NIV was detected in 43 samples (80%). ZEN was not detected in any of the samples tested. The incidence of DON was 19% for Korean beers and 50% for imported beers, whereas the incidence of NIV was 85% for Korean beers and 58% for imported beers. This is the first report that the beers consumed in Korea are contaminated with DON and NIV.

Topics: Beer; Food Contamination; Fusarium; Gas Chromatography-Mass Spectrometry; Humans; Korea; Mycotoxins; Trichothecenes; Zearalenone

A survey was carried out to obtain data on the occurrence of Fusarium mycotoxin in wheat and flour samples collected from local markets in Egypt and to study the influence of gamma-irradiation on controlling the occurrence of these mycotoxins in wheat, flour and bread. Deoxynivalenol (DON) was detected in five samples of wheat at levels ranging from 103 to 287 micrograms/kg and one sample each of flour and bread at concentrations 188 and 179 micrograms/kg. Zearalenone (ZEN) was detected in ten samples of wheat at levels from 28 to 42 micrograms/kg and four samples each of flour and bread at concentrations of 95 and 34 micrograms/kg, respectively. T-2 toxin was detected only in one sample each of wheat, flour and bread at concentrations of 2.9, 2.2 and 2.3 micrograms/kg, respectively. Gamma-irradiation at dose level of 6 kGy completely eliminated fungal flora in flour and wheat. DON, ZEN and T-2 toxin concentrations are reduced to 85, 20 and 2.0 micrograms/kg for wheat and to 125, 45 and 1.0 micrograms/kg for flour after 4 kGy exposure and a sharp drop in Fusarium toxin levels occurred at 6 kGy and as eliminated at 8 kGy. Bread prepared from 6 kGy was contaminated with Fusarium toxin at levels below 5 microgram/kg. It was noticed that gamma-irradiation reduce greatly the natural occurrence of Fusarium mycotoxins in bread.

Topics: Bread; Egypt; Flour; Food Irradiation; Food Microbiology; Fusarium; Gamma Rays; Mycotoxins; T-2 Toxin; Trichothecenes; Triticum; Zearalenone

In Argentina, there is rather little information about the natural occurrence of mycotoxins in feedstuffs. The aim of this work was to determine the fungal flora and natural incidence of aflatoxin B1 (AFB1), zearalenone (ZEA) and deoxynivalenol (DON) in poultry feeds from 5 factories of Río Cuarto, Córdoba. Three hundred samples were taken from May 1995 to May 1996. Fungal counts of poultry feeds ranged 10(4) to 10(6) CFU g-1. The lowest counts were obtained on the first months from the sampling (May to September 1995) with mean values significantly different from those found at the last of the sampling (October 1995 to April 1996). The most prevalent species isolated of poultry feed samples belonged to the genera Penicillium that was present in 98% of the samples, Fusarium (87%) and Aspergillus (52%). Fusarium species isolated were: F moniliforme in 73% of the samples, F subglutinans (35%), F graminearum (20%) and within Aspergillus species: A. parasiticus (33%) and A. flavus (8%) were identified. In poultry feeds aflatoxin B1 (AFB1) was the most significant mycotoxin with levels ranging from 17 to 197 ng/g. For deoxynivalenol (DON) the levels ranged from 240 to 410 ng/g. Only three out of 300 samples were contaminated with zearalenone (ZEA) in concentrations of 30, 120 and 280 ng/g. These are preliminary data on this subject in our region.

Topics: Aflatoxin B1; Animal Feed; Animals; Argentina; Colony Count, Microbial; Food Contamination; Food Microbiology; Fungi; Poultry; Seasons; Trichothecenes; Zearalenone

Mycotoxin production (deoxynivalenol (DON), acetyl deoxynivalenol (A DON) and zearalenone) by Fusarium culmorum inoculated on to maize (heat sterilized, irradiation sterilized and non-sterile) and irradiated to 1 kGy or 3 kGy, or unirradiated, was investigated over a period of time. Lowest mycotoxin production was observed on non-sterile maize which may be due to the presence of a competitive microflora on non-sterile maize which may be due to the presence of a competitive microflora on non-sterile maize. In general, mycotoxin production was higher on heat-sterilized grain as compared to irradiation-sterilized maize. It was suggested that this pattern of mycotoxin production was possibly caused by changes in the grain brought about by autoclaving, which favoured mycotoxin production and possibly induced changes in irradiation-sterilized maize which inhibited mycotoxin production. On sterile maize, there was no significant difference in DON production by unirradiated, 1 kGy and 3 kGy irradiated cultures up to 56 d of incubation; between days 56 and 77 of incubation, DON production increased rapidly with largest increases occurring in irradiated (1 kGy and 3 kGy) cultures. On non-sterile grain, neither DON nor A DON were detected in unirradiated cultures of F. culmorum but were detected in cultures irradiated to 1 kGy and 3 kGy. In practice grain should be stored under conditions of temperature and moisture content which prevent fungal growth. However, in this study, the grain was stored under conditions that were approaching ideal for growth of the test organism. The results highlight that irradiation disinfestation of grain must be combined with good grain handling practices so that excessive mycotoxin production can be prevented during storage.

Topics: Fusarium; Gamma Rays; Mycotoxins; Trichothecenes; Zea mays; Zearalenone

The occurrence of deoxynivalenol (DON) and zearalenone in 495 samples of food wheat, barley and rye collected from different regions of Russia, and in 633 samples of fresh harvested wheat from Krasnodar region in 1993 and 1994 was surveyed. DON was detected in 97% of fresh harvested wheat samples in 1993, exceeding maximum tolerated level MTL) in 69% of samples analyzed. 23% of fresh harvested wheat samples were positive for DON and only in 5% of samples investigated DON concentration exceed MTL in 1994. Zearalenone was found in low concentrations in 3 of 154 wheat samples analyzed. Surveys of food wheat, barley and rye samples have shown that 23%, 7% and 3% of lots were positive for DON, respectively, in 1993. DON concentration exceed MTL in 14% of food wheat samples. The frequency of DON contamination of the 1994 food wheat samples was 6%. No mycotoxins were found in the 1994 food barley and rye samples analyzed. Almost all DON contaminated lots of food grain were collected from North Caucasus region.

Topics: Edible Grain; Fusarium; Hordeum; Maximum Allowable Concentration; Mycotoxins; Russia; Secale; Trichothecenes; Triticum; Zearalenone

100 samples of rye and 101 samples of wheat coming out of both conventional and alternative or ecological production were investigated for contamination with mycotoxins with interest for our degree of latitude. Deoxynivalenol (DON) was found with thin-layer-chromatography in 131 of 201 samples altogether. A top level of 1250 micrograms DON kg-1 in rye of alternative offspring was detected. The average burden in contaminated rye coming from ecological production was 427 micrograms kg-1 and a mean level of 160 micrograms kg-1 resulting in rye out of conventional growth conditions. In wheat, conventionally grown yield showed slightly lower contamination (mean levels of 420 micrograms DON kg-1 towards 486 micrograms kg-1). The toxins 3-acetyl-deoxynivalenol, nivalenol and fusarenone X were detected in some samples by thin-layer-chromatography. This results could not be confirmed by gas chromatography -mass spectrometry. Zearalenone was found in 40 out of the number of 201 samples of grain by HPLC with fluorescence detection. An average of 6 micrograms and 24 micrograms zearalenone kg-1 in conventionally and alternatively grown wheat and 4 micrograms and 51 micrograms zearalenone kg-1 in conventionally and alternatively produced rye was detected. The highest finding of zearalenone was 199 micrograms kg-1 in alternatively grown rye. Skin toxicity testing did not show any reference of contamination with type-A-trichothecenes. No correlation between contamination of zearalenone or deoxynivalenol and thousand-kernel-weight was detected.

Topics: Agriculture; Chromatography, Thin Layer; Food Microbiology; Gas Chromatography-Mass Spectrometry; Humans; Mycotoxins; Secale; Skin; Skin Tests; Trichothecenes; Triticum; Zearalenone

The Fusarium mycotoxin zearalenone (ZEA), added at a level of 2 micrograms/ml, was reduced stereoselectively by cultures of Candida tropicalis, Torulaspora delbrückii, Zygosaccharomyces rouxii, and 7 Saccharomyces strains to both alpha- and beta-zearalenol. In contrast, only alpha-zearalenol was produced from ZEA by Pichia fermentans and several yeast strains of the genera Candida, Hansenula, Brettanomyces, Schizosaccharomyces, and Saccharomycopsis. No glucose conjugates of ZEA (zearalenone-4-beta-D-glucopyranoside) were detected. The trichothecene mycotoxin deoxynivalenol (DON) was not metabolized by any of the yeast strains that were used for analysis.

Topics: Beer; Biotransformation; Species Specificity; Time Factors; Trichothecenes; Wine; Yeasts; Zearalenone

Mycotoxins zearalenone and vomitoxin (4-deoxynivalenol) are often joint contaminants of grains infested by micromycetes of the genus Fusarium. Toxic effects of both mycotoxins on experimental organisms and farm animals are well known, but we have not found any literary reference to toxic effects of the combination zearalenone and vomitoxin. Embryotoxic effects of zearalenone, vomitoxin and combinations of various doses of zearalenone with constant addition of vomitoxin were studied in a three-day chick embryo. The objective of the study was to determine the coaction of vomitoxin on zearalenone embryotoxicity. Thermostat-incubated fertile eggs of White Leghorn hens were candled after three-day incubation, the shell above the embryo was removed, and within the embryotoxicity range zearalenone, vomitoxin and various doses of zearalenone with constant addition of 2 micrograms vomitoxin were applied to morphologically normal embryos. The groups of ten embryos were applied mycotoxins and their combinations in 10 microliters of their solutions to amnions using a special glass micropipette. Control group comprised twenty embryos which were applied 10 microliters of solvents used, 1% NaHCO3 and 10% ethanol. The eggs were covered with glass plates and their incubation was going on until the eighth day of their development. The embryos that died during incubation were discarded. On the eighth day of development, surviving embryos were taken out from the eggs and malformations of head, orofacial region, body wall, limbs and heart were determined microscopically. Tab. I shows total numbers of dead and malformed embryos after application of the particular doses of zearalenone, vomitoxin, their combinations and control solvents. The embryotoxicity range started at a dose of 5 to 20 micrograms per embryo. Zearalanone did not have any teratogenic effects on chick embryos. Applications of high doses of zearalenone (100 and 30 micrograms) instantly caused arrhythmia, atrio-ventricular dissociation or even heart stoppage. The beginning of the embryotoxicity range for vomitoxin was found to be within the narrow range of 1 to 3 micrograms per embryo. Among malformations, only a defect of the interventricular septum of the heart was found in 4% of the cases. The combined embryotoxic effects of zearalenone and vomitoxin were of additive, and mostly embryolethal nature. Among the malformations searched for, only 5% of the embryos exhibited a defect of the interventricular septum

Topics: Abnormalities, Drug-Induced; Animals; Chick Embryo; Mycotoxins; Trichothecenes; Zearalenone

The effects of low dietary concentrations of Fusarium mycotoxins (deoxynivalenol (DON), 15-acetyl-DON, and zearalenone) on growth, immunological, and hematological parameters were determined in young pigs during a 28-day feeding experiment. Clean and naturally contaminated corn were incorporated into basal diets formulated to contain 0.00, 0.75, 1.50, and 3.00 mg DON/kg diet. A pair-fed control animal was used for comparison with each animal receiving the highest level of contamination (diet 4). Skin temperature, measured during the first week of the experiment, decreased linearly as the dietary mycotoxin concentration increased. Several other linear effects were observed: depressed feed intake throughout the experiment, reduction in thyroid size (absolute/relative), and changes in the appearance of the esophageal region of the stomach (thicker and higher degree of folding with increasing toxin concentration). Serum T4 (thyroxine) levels increased quadratically after 7 and 28 days of exposure compared to control animals. This change coincided with an increase in albumin levels, a decrease in alpha-globulin levels, and an overall increase in albumin/globulin ratio as the level of contamination increased. After immunization with sheep red blood cells (SRBC), animals fed contaminated diets showed a delayed response in peak titers. At the end of the experiment an increase in the segmented neutrophil count was observed. The following observations were made for animals consuming diet 4 as compared to the pair-fed controls: lower skin temperature, better feed efficiency, more corrugated stomachs, reduced alpha-globulin levels, and lower antibody titers to SRBC.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animal Feed; Animals; Body Weight; Fusarium; Hematopoiesis; Immune System; Mycotoxins; Organ Size; Swine; Trichothecenes; Zearalenone

A preliminary survey for the natural occurrence of Fusarium mycotoxins--nivalenol (NIV), deoxynivalenol (DON) and zearalenone (ZEN)--in cereals imported mostly from Australia to Papua, New Guinea (PNG) in 1991 was carried out by TLC, GLC, and HPLC analysis. The data have revealed various kinds of cereals and cereal flours, such as wholemeal flour, millet meal, corn flour, brown trukai rice, and others, were significantly contaminated with these mycotoxins in various concentrations. Among these Fusarium mycotoxins, the level of ZEN was surprisingly high. This is the first report on the natural occurrence of Fusarium mycotoxins in PNG cereals.

Topics: Chromatography, Gas; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Edible Grain; Food Contamination; Food Microbiology; Fusarium; Mycotoxins; New Guinea; Trichothecenes; Zearalenone

The biotransformation of the Fusarium mycotoxins deoxynivalenol and zearalenone by the normal bacterial gut flora of pigs was examined in this in vitro study. For that purpose, suspensions of intestinal contents (duodenum, jejunum, caecum, colon, rectum) of porcine origin were incubated anaerobically with deoxynivalenol (DON) or zearalenone (ZEA). DON and ZEA were degraded by the flora of the caudal segments (caecum, colon, rectum) of the gut--particularly the colon content--whereas the microorganisms of the cranial segments (duodenum, jejunum) exhibited no transforming activity. DON was showed to be deepoxidated, ZEA was hydrolyzed to alpha-zearalenol and an unknown metabolite. The transformation of DON was correlated with a loss of cytotoxicity, which could be demonstrated in the MTT(3-[4,5-dimethylthiazol-2yl]-2,5-diphenyltetrazolium++ + bromide)-cell-culture assay using swine kidney cells as target cells. The results of the study presented here correspond with the data found in in vivo studies. On the basis of these findings one could conclude that this in vitro method seems to be well suited to the study of the transformation of mycotoxins by the microflora of the gut. The in vitro study is cheaper than a feeding trial, and the preliminary information on the metabolism of mycotoxins obtained in such studies is helpful in designing feeding trials more clearly. Besides the simple and fast handling, reproducibility and the protection of the animals studied are further advantages of this in vitro method. In connection with the MTT-cell-culture assay, additional information about the cytotoxic potential of the bacterial transformation products can be obtained.

Topics: Animals; Biotransformation; Cells, Cultured; Chromatography, High Pressure Liquid; Gastrointestinal Contents; Intestinal Mucosa; Intestines; Swine; Trichothecenes; Zearalenone

The incidence of Fusarium toxins-trichothecenes and zearalenone in 100 samples of whole grains as well as their distribution in the by-products (arising of 32 whole corn samples) of the wet-grinding of corn were studied, the samples coming from the central and northern areas of the Santa Fe Province, Argentina. The analyses were carried out by thin-layer chromatography. A 33% of the samples was found to be contaminated by deoxynivalenol (DON), its amount ranging from traces to 1,200 micrograms/Kg; 15% of the samples contained T-2 toxin from 900 to 2,400 micrograms/Kg and only traces of zearalenone were detected in one sample. Only few samples (7%) included diacetoxiscirpenol (DAS), nivaleno (NIV) and neosolaniol (NS) and 7% of the samples evidenced contamination by more than one mycotoxin. It is worth noticing that DON was found to be contaminating only those by-products destined to human consumption. Conversely, T-2 toxin was found in by-products destined to both human and animal consumption.

Topics: Food Contamination; Food Microbiology; Food Technology; Fusarium; Mycotoxins; Trichothecenes; Zea mays; Zearalenone

Fifty-five Fusarium isolates belonging to nine species were collected from fungus-invaded tissue of stored sugar beets and identified as F. acuminatum (11 isolates), F. avenaceum (1 isolate), F. culmorum (1 isolate), F. equiseti (23 isolates), F. graminearum (4 isolates), F. oxysporum (1 isolate), F. solani (4 isolates), F. sporotrichioides (7 isolates), and F. subglutinans (2 isolates). All isolates were cultured on autoclaved rice grains and assayed for toxicity by feeding weanling female rats the ground-rice cultures of the isolates in a 50% mixture with a regular diet for 5 days. Fifty-eight percent of the isolates were acutely toxic to rats, 26% caused hematuria, 18% caused hemorrhages, and 29% caused uterine enlargement. In most cases, toxicity could not be accounted for by the known toxins found. The following mycotoxins were found in extracts of the rice cultures: zearalenone (22 to 6,282 micrograms/g), chlamydosporol (HM-8) (68 to 4,708 micrograms/g), moniliformin (45 to 400 micrograms/g), deoxynivalenol (10 to 34 micrograms/g), 15-acetyldeoxynivalenol (5 to 10 micrograms/g), diacetoxyscirpenol (22 to 63 micrograms/g), monoacetoxyscirpenol (21 to 26 micrograms/g), scirpenetriol (24 micrograms/g), T-2 toxin (4 to 425 micrograms/g), HT-2 toxin (2 to 284 micrograms/g), neosolaniol (2 to 250 micrograms/g), and T-2 tetraol (4 to 12 micrograms/g). F. equiseti was the predominant species found on visibly molded beets in the field. Six of 25 moldy sugar beet root samples collected in the field contained zearalenone in concentrations ranging between 12 and 391 ng/g, whereas 10 samples from commercial stockpiles were negative for zearalenone.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Female; Fusarium; Hematuria; Hemorrhage; Mycotoxins; Plants, Edible; Rats; Rats, Sprague-Dawley; Trichothecenes; Zearalenone

Polyclonal antisera against zearalenone (ZEA) were produced in rabbits after immunization with ZEA-oxime coupled to human serum albumin. Using these antibodies and a ZEA-oxime-horseradish peroxidase conjugate in a competitive direct enzyme immunoassay (EIA), the detection limit for ZEA was 70 pg/ml. The relative cross-reactivities of the assay with ZEA, alpha-zearalenol, beta-zearalenol, zearalanone, alpha-zearalanol, and beta-zearalanol, respectively, were 100%, 37.3%, 7.2%, 59.2%, 5.3%, and 3.9%, respectively. This EIA and two EIAs for deoxynivalenol (DON) and 3-acetyldeoxynivalenol(3-AcDON) (Usleber et al., 1991) were used to analyze wheat samples. The limits of determination for DON, 3-AcDON, and ZEA in wheat were 200 ppb, 50 ppb, and 20 ppb, respectively. The analysis of reference materials (wheat flour) containing DON by EIA showed good agreement with the nominal values. The EIA for ZEA was in addition used to analyze biological fluids, obtained during a feeding trial. Two lactating cows were administered 25 mg and 100 mg ZEA per day, respectively, over a period of 6 days. Serum, milk, urine, and feces were assayed in the ZEA-EIA with and without sample treatment with beta-glucuronidase prior to the analysis. Maximum toxin levels (ZEA-equivalents) found in milk were 0.4 and 1.2 ppb (glucuronides). The toxin concentration in milk decreased rapidly after the last toxin administration. In the urine, maximum levels of toxin-glucuronide conjugates were 23 ppb and 24 ppb, respectively. The serum toxin levels corresponded to those found in milk. In the feces, mean values were 150 ppb and 500 ppb, respectively, no conjugated toxins were found in feces.

Topics: Animals; Cattle; Edible Grain; Female; Fusarium; Immunoenzyme Techniques; Trichothecenes; Zearalenone

A total of 82 feed manufacturers located within seven midwestern states (Iowa, Nebraska, Minnesota, Illinois, Indiana, Ohio, Michigan) participated in a survey of mold and mycotoxin contamination of corn. Samples were submitted from a composite of the grading samples taken from each incoming load of corn. The survey was initiated in July 1988. During the 12-mo period, moisture content of the corn samples upon receipt at the laboratory ranged from 10.5 to 13.3%. The greatest variation occurred in the springtime. Iowa's corn samples were driest (11.2%), and samples submitted from Ohio were wettest (12.8%). Mold counts averaged 2.63 x 10(4) per gram during the year. The predominant mold found was Fusarium sp. Samples were checked by black light and averaged 25.4% positive during the period. When assayed for mycotoxins, 19.5% of the samples were positive for at least one of the following: aflatoxin, zearalenone, T2 toxin and deoxynivalenol (vomitoxin). Aflatoxin and T2 toxin made up the majority of these samples containing toxin. The highest incidence of mycotoxin-contaminated corn (48%) occurred in samples submitted in July of 1988. Over the 12-mo period, the highest mycotoxin contamination occurred in Iowa, Illinois and Michigan. When samples were subjected to 90% relative humidity and 32 degrees C, an average of 3.9 d was required for mold growth to appear. After incubation, 24.7% of the samples contained one of the four toxins. The data indicate that mold and mycotoxin contamination of mixed samples of corn is widespread, even in the midwestern corn belt of the U.S.

Topics: Aflatoxins; Animal Feed; Animals; Aspergillus; Colony Count, Microbial; Food Microbiology; Fungi; Fusarium; Midwestern United States; Mycotoxins; Penicillium; Seasons; T-2 Toxin; Trichothecenes; Zea mays; Zearalenone

Forty Fusarium isolates obtained from maize fields were screened for moniliformin production on maize kernels. Twelve isolates, including seven of F. subglutinans, were found to produce moniliformin at levels ranging from 0.4 to 64 ppm. Twenty six isolates were also screened for production of deoxynivalenol, diacetoxyscirpenol, T-2 toxin and zearalenone. Of these, 22, including all 11 isolates of F. graminearum, produced zearalenone at levels ranging from 0.1 to 96.0 ppm, while 13 produced T-2 toxin at low levels, (less than 1.1 ppm). Deoxynivalenol and diacetoxyscirpenol were each produced by six isolates, also at low levels (less than 1.0 ppm). Three isolates of F. graminearum and one of F. sambucinum produced four toxins simultaneously.

Topics: Cyclobutanes; Fusarium; Mycotoxins; New Zealand; T-2 Toxin; Trichothecenes; Zea mays; Zearalenone

To assess the potential for mycotoxin contamination of the human food supply following the 1988 U.S. drought, 92 grain food samples were purchased from retail outlets in the summer of 1989 and surveyed for aflatoxin B1, zearalenone, and deoxynivalenol (DON [vomitoxin]) by monoclonal antibody-based competitive enzyme-linked immunosorbent assay (ELISA). Only one sample (buckwheat flour) was found to contain aflatoxin B1 (12 ng/g), whereas zearalenone was found in 26% of the samples at a mean concentration of 19 ng/g. In contrast, the DON ELISA was positive in 50% of the samples at a detection level of 1.0 micrograms/g. Between 63 and 88% of corn cereals, wheat flour/muffin mixes, rice cereals, and corn meal/muffin mixes yielded positive results for DON, whereas 25 to 50% of oat cereals, wheat- and oat-based cookies/crackers, corn chips, popcorn, and mixed-grain cereals were positive for DON. The mean DON content of the positive samples was 4.0 micrograms/g, and the minimum and maximum levels were 1.2 and 19 micrograms/g, respectively. When positive ELISA samples were also analyzed by high-performance liquid chromatography, a strong correlation between the two methods was found. The presence of DON in the two highest samples, corn meal and mixed-grain cereal, which contained 19 and 16 micrograms/g, respectively, was quantitatively confirmed by gas chromatography-mass spectrometry. The results indicated that DON was present in 1989 retail food products at concentrations that exceeded those found in previous market surveys and that have been experimentally associated with impaired animal health.

Topics: Aflatoxin B1; Aflatoxins; Edible Grain; Enzyme-Linked Immunosorbent Assay; Food Contamination; Trichothecenes; Zearalenone

A survey for the natural occurrence of Fusarium mycotoxins, deoxynivalenol (DON), nivalenol (NIV) and zearalenone (ZEN), in Dutch cereals (totaling 29 samples) harvested in 1984/1985, showed that 90%, 79% and 62% of samples were contaminated with DON, NIV and ZEN, respectively. Average contents (ng/g) in the total of positive samples were 221 (DON), 123 (NIV) and 61 (ZEN). Among the cereals examined, the highest concentrations (ng/g) was 3198 (DON), 1875 (NIV) and 677 (ZEN) in a yellow corn sample for animal feed. The results of this survey show that Dutch cereals were relatively significantly contaminated with Fusarium mycotoxins.

Topics: Edible Grain; Food Contamination; Fusarium; Hordeum; Mycotoxins; Netherlands; Resorcinols; Secale; Sesquiterpenes; Trichothecenes; Triticum; Zea mays; Zearalenone

Ninety five samples of various Egyptian feedstuffs were investigated for the aflatoxin B1, B2, G1, and G2 means thin layer chromatography (TLC). Out of these samples 44.2% were positive (maize, rice crack, rice germ, rice germ cake, rice bran, wheat bran, cotton seed, cotton seed cake, peanut, and mixed feed for broilers, egg production, calf fattening and milk production). High percentage (90.5%) of the positive samples were contaminated with less than 100 ppb total aflatoxins. Peanut from "Ismailia" showed the highest contamination-mean of 400 ppb aflatoxin B1. The contamination relationship between kernels and shell of the same pods of the peanut was 1:7. The lowest contamination-mean was 5 ppb B1 in soya bean samples. All samples of horse bean and fish meal were negative. Aflatoxin B1 was present alone so frequently (in 76.2% of the positive samples). The relationship between the concentrations of aflatoxins B2:G1:B1 was 1:2.3:22.4. 51 different samples of foods and feeds from various Egyptian regions were collected and investigated for the nephrotoxic mycotoxin ochratoxin A means TLC. Twelve samples (23.5%) from them were designated as positive samples. The positive samples belonged to white maize, wheat, wheat bran, beans, rice germ, rice germ cake, broilers feed, egg production feed, and milk production feed; whereas the yellow maize (hybrid), soya beans, wheat soya meal, rice crack, cotton seed, cotton seed cake, and fish meal samples were negative. The contamination range was from 4 ppb to 577 ppb with an average of 58.2 +/- 22.9 ppb. Half of the positive samples was contaminated with 10-100 ppb whereas 41.7% from the positive samples had less than 10 ppb and 8.3% only had more than 100 ppb. Citrinin is existing in Egyptian food and feedstuffs. Out of 52 different samples--from various Egyptian regions-15.4% were positive. These were rice bran, rice germ, maize (white), wheat bran, cotton seed cake and fish meal. The highest contamination was in fish meal (40-70 ppb) whereas the lowest was in wheat bran (3 ppb). Mean of the contamination level was 25.9 +/- 3.4 ppb with a range of 3-70 ppb. For the first time in the Egyptian foods and feeds will be informed about the presence of the mycotoxin zearalenone with a high concentration. From several Egyptian places, 64 samples were collected (4 samples for each food or feed stuff).(ABSTRACT TRUNCATED AT 400 WORDS)

Topics: Aflatoxins; Animal Feed; Animals; Chromatography, Thin Layer; Citrinin; Egypt; Food Contamination; Mycotoxins; Ochratoxins; Trichothecenes; Zearalenone

Topics: Abortion, Veterinary; Animal Feed; Animals; Cohort Studies; Female; Fetal Death; Food Contamination; Infertility; Litter Size; Mycotoxins; Pregnancy; Prospective Studies; Swine; Swine Diseases; Trichothecenes; Zearalenone

A total of 399 indigenous Fusarium strains mainly isolated from silage maize were tested for the production of zearalenone and type A trichothecenes by thin-layer chromatography and biological assays. About 45% of the isolates examined were capable of producing different levels of zearalenone and trichothecene toxins on a cracked corn substrate. The majority of these strains (75%) produced zearalenone only and no trichothecenes type A. The results of the biological tests indicated a higher rate of toxin-positive extracts than chemical analysis. Isolates of nine out of seventeen Fusarium species examined produced one or several mycotoxins looked for. The most important toxin producers were F. culmorum and F. crookwellense (zearalenone) and F. sporotrichioides (trichothecenes type A), respectively. F. avenaceum, the species most frequently isolated from silage maize, produced neither zearalenone nor trichothecenes but avenacein Y a antibiotic compound. First results of a study of the production of type B trichothecenes have shown that indigenous F. culmorum isolates were capable of producing high levels of deoxynivalenol.

Topics: Anti-Bacterial Agents; Benzopyrans; Food Microbiology; Fusarium; Poaceae; Silage; Trichothecenes; Zea mays; Zearalenone

A toxicogenic strain of Fusarium graminearum which produces DON and ZEA was cultivated on natural solid substrates (wheat, polished rice and hulled rice) under different environmental conditions. The production of both toxins and mycelium growth (in terms of glucosamine) were evaluated to establish the relation between the production of DON and ZEA and the different mycelium growth on the substrates mentioned above. Polished rice was the substrate on which most production of both toxins was obtained. Comparing the three substrates studied, the highest quantities of DON were obtained at a temperature of 27 degrees C during incubation period, being indifferent to the presence of light except in the case of hulled rice. Whereas for ZEA the best conditions in wheat and polished rice were medium temperatures (17 degrees-21 degrees C respectively) and darkness. While in hulled rice the ideal conditions for the production of both toxins were temperature of 27 degrees C and the presence of light. Concerning the mycelium growth, this was very scarce when cultivated in hulled rice, increasing in polished rice and being largest in wheat. The increase or reduction of the mycelium growth in the different substrates was not proportional to the increase or decrease of the production of both toxins. Therefore, production of DON and ZEA could be subjected to the nature of the substrate and environmental conditions, more than the rate of development of Fusarium graminearum in cereal grains.

Topics: Food Microbiology; Fusarium; Microbiological Techniques; Oryza; Trichothecenes; Triticum; Zearalenone

Bavarian cereals and wheat flour from the 1987 harvest were analysed for nivalenol (NIV) and deoxynivalenol (DON) using high performance liquid chromatography (HPLC) and for T-2 toxin and zearalenone (ZEA) by enzyme-linked-immunosorbent assay (ELISA). The study included 190 field samples of wheat, barley, rye and oat with visibly damaged ears, 45 samples of wheat intended for feed production and two series of wheat flour (type 550) and whole wheat flour collected in October 1987 and June 1988. The field samples examined showed a high DON contamination of wheat (87%) with an average of 3.96 mg/kg and a maximum of 43.8 mg/kg. Mean levels between 0.33 mg/kg and 0.27 mg/kg DON could be detected in barley, rye and oat. Of the wheat samples, 58% contained ZEA with a maximum of 1.560 mg/kg. The highest levels of ZEA were detected in samples which also showed high concentrations of DON. The NIV and T-2 toxin levels were comparatively low. Thirty percent of the samples showed NIV concentrations between 0.04 mg/kg and 0.29 mg/kg and 38% contained between 0.005 and 0.60 mg/kg of T-2. In the wheat samples for feed production, only DON was detected with an average of 0.190 mg/kg and a maximum of 0.75 mg/kg. The highest DON levels (0.58 mg/kg) from October 1987 were found in the wheat flour samples which were lower than the highest DON concentration (3.24 mg/kg) detected in the samples collected during June 1988. This fact was probably due to a substantial amount of non-contaminated wheat from 1986. The toxin concentrations in the whole wheat flour were not higher than in the type 550 flour.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animal Feed; Animals; Edible Grain; Flour; Food Contamination; Food Microbiology; Fusarium; Germany, West; Humans; Mycotoxins; T-2 Toxin; Trichothecenes; Zearalenone

An investigation for the occurrence of nivalenol (NIV), deoxynivalenol (DON) and zearalenone (ZEN) in cereals (ten wheat, one rye and one corn) harvested in Canada have been carried out using a procedure, which is rapid and sensitive for Fusarium mycotoxins. NIV, DON and ZEN were detected in 4, 9 and 9 out of ten wheat samples, and their average levels in the positives were 23 ng/g, 1257 ng/g and 9 ng/g, respectively. One rye and one corn were also contaminated with a minor amount of NIV. This is the first evidence for the natural occurrence of NIV in cereals grown in Canada, though its level was far less than DON.

Topics: Canada; Edible Grain; Food Contamination; Fusarium; Resorcinols; Secale; Sesquiterpenes; Trichothecenes; Triticum; Zea mays; Zearalenone

Five Fusarium species were isolated from the grain of dent corn (Zea mays) selected from 20 of 32 damaged fields in 10 counties in Minnesota on the basis of hyphal growth visible on kernels in the field. Three mycotoxins were identified in the infected ears: zearalenone, deoxynivalenol, and 15-acetyl-deoxynivalenol. This is the first report of the presence of 15-acetyl-deoxynivalenol on corn ears in the field prior to harvest and in combination with deoxynivalenol and zearalenone. Ninety-nine cultures were selected from colonies growing from kernels on an agar medium; 30% of the cultures were F. graminearum, 23% were F. subglutinans, 20% were F. moniliforme, 14% were F. oxysporum, and 12% were F. proliferatum.

Topics: Fusarium; Minnesota; Mycotoxins; Trichothecenes; Zea mays; Zearalenone

This is the first report of the natural coexistence of a group of Fusarium mycotoxins (nivalenol [NIV], deoxynivalenol [DON], 3-acetyl-deoxynivalenol [3-ADON], 15-acetyl-deoxynivalenol [15-ADON], and zearalenone [ZEN]) in corn from Linxian, China, an area with a high risk of esophageal cancer. Using thin layer chromatography (TLC), high pressure liquid chromatography (HPLC), and gas chromatography (GC), 107 corn samples from Linxian were analyzed. The average levels of NIV and DON were 757 +/- 707 (54-2,760) ng/g and 5,376 +/- 4,460 (360-12,670) ng/g, respectively, with 100% positivity in 24 corn samples consumed as staple food by esophageal cancer patients and their families. Other corn samples collected from five villages in Linxian at different seasons in 1984-1986 also revealed high levels of NIV and DON contamination, with 100% positivity, suggesting that they are consistently and widely present in corn in that area. Levels of 3-ADON and 15-ADON in Linxian corn were 113 +/- 57 and 495 +/- 538 ng/g, respectively. Crude extracts of corn samples collected from esophageal cancer patients' families and the HPLC-purified NIV and DON fractions induced significant chromosome aberrations in V79 cells. Pure toxins of NIV, DON, T-2, and 3-ADON also induced chromosome aberrations in V79 cells at very low concentrations (ng levels/ml medium). Cytotoxic effects were observed at slightly higher concentrations. The levels and kinds of trichothecenes were in positive co-relation with the incidence of esophageal cancer. The data suggest that trichothecenes in food may possibly be associated with esophagitis and esophageal cancer in Linxian.

Topics: China; Dose-Response Relationship, Drug; Esophageal Neoplasms; Fusarium; Humans; Mutagenicity Tests; Mycotoxins; Resorcinols; Sesquiterpenes; Trichothecenes; Zea mays; Zearalenone

The effect that dietary exposure to the naturally-occurring Fusarium graminearum toxins deoxynivalenol (DON) and zearalenone (ZEA) may have on immune function was assessed in the B6C3F1 mouse. Dietary DON depressed the plaque-forming response to sheep red blood cells, the delayed hypersensitivity response to keyhole limpet haemocyanin and the ability to resist Listeria monocytogenes. Listerial resistance was similarly decreased in control mice fed restricted diets comparable to the dietary restriction caused by DON-induced feed refusal, whereas equivalent food restriction did not decrease the plaque or delayed hypersensitivity responses. ZEA ingestion decreased resistance to L. monocytogenes but did not affect splenic plaque-forming or delayed hypersensitivity responses. Resistance to Listeria was reduced to a greater extent by co-administration of DON and ZEA than by DON alone, whereas the ability of DON to inhibit the delayed hypersensitivity response was significantly lessened in the presence of ZEA. While effects on resistance to Listeria and delayed hypersensitivity were detectable in mice ingesting the mycotoxins for 2-3 wk, these effects disappeared upon extension of the feeding period to 8 wk. In contrast, some effect on the plaque-forming response was detectable with both the 2- and the 8-wk period of mycotoxin ingestion. Immunosuppression can thus result from ingestion of F. graminearum-infected agricultural staples, the suppression being attributable to interactions between direct immunotoxic effects of DON and ZEA and nutritional effects associated with DON-induced food refusal.

Topics: Animals; Diet; Female; Hemolytic Plaque Technique; Hypersensitivity, Delayed; Immunity; Immunity, Innate; Listeriosis; Mice; Resorcinols; Sesquiterpenes; Time Factors; Trichothecenes; Zearalenone

Weanling female B6C3F1 mice were fed semi-purified diets containing 0, 0.5, 2.0, 5.0, 10.0 or 25.0 ppm (mg/kg) deoxynivalenol (DON) over 8 wk and were assessed for effects on feed intake, body-weight gain, terminal organ weights, histopathology, haematology and serum immunoglobulin levels. To determine whether DON effects were potentiated by the oestrogen zearalenone (ZEA), a mycotoxin frequently found to occur with DON in cereals, two additional groups of mice were fed diets containing either 10 ppm ZEA or 10 ppm ZEA plus 5 ppm DON. The rate of body-weight gain was significantly reduced (P less than 0.01) for all mice consuming feed containing 2.0 ppm or more of DON, whereas only the mice ingesting the diet containing 25 ppm DON showed a significantly decreased (P less than 0.01) rate of feed consumption. Gross and histopathological evaluation of thymus, spleen, liver, kidney, uterus, small intestine, colon, heart, brain, lungs and bone marrow from control and all mycotoxin-exposed mice revealed that these tissues were normal in appearance and in histological architecture. DON-amended diets did however, cause dose-dependent decreases in the terminal organ weights recorded (thymus, spleen, liver, kidney and brain). In the DON-treated groups, statistically significant dose-dependent decreases in the counts of total circulating white blood cells were associated with an increase in polymorphonuclear neutrophils and a decrease in lymphocytes and monocytes. Dietary DON caused a dose-dependent decrease in serum IgM but, in contrast, a dose-dependent increase in serum IgA. In none of the above instances was 10 ppm ZEA shown to act synergistically or antagonistically with 5 ppm DON. Since dietary DON at levels as low as 2.0 ppm exerted significant effects on the growing B6C3F1 female mouse, future approaches should include studies of the mechanisms by which this mycotoxin affects nutrient utilization and modifies the normal immune response.

Topics: Animals; Body Weight; Diet; Dose-Response Relationship, Drug; Eating; Erythrocyte Count; Female; Immunoglobulins; Kidney; Leukocyte Count; Liver; Mice; Mice, Inbred Strains; Organ Size; Resorcinols; Sesquiterpenes; Trichothecenes; Zearalenone

Two samples of "refusal factor" corn, one stored frozen in Minnesota and one stored dry in Indiana since 1972 or 1973, were analyzed for the presence of Fusarium spp. and Fusarium toxins. Both samples were from corn refused by swine in Indiana from 1972 to 1973. Sample FS 808 (stored in Indiana) contained 20 ppm of deoxynivalenol (20 micrograms/g), 16 ppm of 15-acetyl-deoxynivalenol, 5 ppm of zearalenone, and 0.2 ppm of alpha-zearalenol. Sample FS 362 (stored in Minnesota) contained 3 ppm of deoxynivalenol, 1 ppm of 15-acetyldeoxynivalenol, and 0.3 ppm of zearalenone. The presence of 15-acetyl-deoxynivalenol is significant because it is the first report of it occurring naturally in refusal factor corn, and it may account in part for the refusal that could not be solely attributed to deoxynivalenol.

Topics: Animal Feed; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Food Contamination; Food Microbiology; Fusarium; Gas Chromatography-Mass Spectrometry; Indiana; Minnesota; Mycotoxins; Time Factors; Trichothecenes; Zea mays; Zearalenone; Zeranol

Cereals, foods and feeds sampled in Taiwan, China and the U.S.S.R. were contaminated with nivalenol, deoxynivalenol and zearalenone. The frequencies and levels of contamination are similar to those observed in the cereals of Japan and Korea. This is the first report on the natural occurrence of nivalenol, deoxynivalenol and zearalenone in Chinese and U.S.S.R. cereals, foods and feeds.

Topics: China; Edible Grain; Flour; Food Contamination; Hordeum; Resorcinols; Sesquiterpenes; Taiwan; Trichothecenes; Triticum; USSR; Zearalenone

The fungal species isolated from Korean cereals (barley, polished barley, wheat, rye, and malt) were Alternaria spp., Aspergillus spp., Chaetomium spp., Drechslera spp., Epicoccum sp., Fusarium spp., and Penicillium spp., etc. The number of Fusarium strains isolated was 36, and their ability to produce Fusarium mycotoxins on rice was tested. Nivalenol (NIV) was produced by Fusarium graminearum (7 of 9 isolates), Fusarium oxysporum (3 of 10 isolates), and Fusarium spp. (7 of 15 isolates). Of 15 isolates of Fusarium spp., 6 formed deoxynivalenol (DON). Fusarenon-X and 3-acetyl-DON were produced by most NIV- and DON-forming isolates, respectively. Zearalenone was produced by 3 isolates of F. graminearum, 1 isolate of Fusarium equiseti, and 11 isolates of Fusarium spp. T-2 toxin was not produced by any Fusarium isolates. The highest concentrations of mycotoxins produced by Fusarium isolates were 77.4 (NIV), 5.3 (DON), 138.3 (fusarenon-X), 40.6 (3-acetyl-DON), and 23.2 (zearalenone) micrograms/g.

Topics: Chaetomium; Edible Grain; Food Microbiology; Fungi; Fusarium; Korea; Mitosporic Fungi; Mycotoxins; Trichothecenes; Zearalenone

The production of deoxynivalenol (DONI) on rice, corn, wheat, and barley grains by Fusarium graminearum Schw. NRRL 5883 was investigated. Highest yields (91.9-202 ppm) were obtained on rice; yields on the other substrates were: corn (34.1-84.5 ppm), wheat (3.6-24.4 ppm), and barley (0-6.6 ppm). Fifty isolates of Fusarium from corn inoculated in the field in 1979 with a mixture of strains of F. graminearum, originally collected from corn plants infected with stalk rot, were tested for DONI production on corn. Twenty of these were also tested for zearalenone production. One isolate produced more than 200 ppm DONI, 13 produced 20-50 ppm, 17 produced 10-20 ppm, and the rest produced less than 10 ppm. The only isolate that did not produce DONI was not identified as F. graminearum. All 20 isolates tested produced zearalenone; 18 produced higher levels of zearalenone (15.4-369 ppm) than of DONI. The other 2 isolates formed essentially the same levels of zearalenone and DONI-37 and 30 ppm, and 15 and 16 ppm, respectively.

Topics: Food Microbiology; Fusarium; Hordeum; Oryza; Resorcinols; Sesquiterpenes; Trichothecenes; Triticum; Zea mays; Zearalenone

A 5-wk feeding trial was conducted with 30 castrated male and 28 female, 5-wk-old crossbred piglets. Three different deoxynivalenol (DON) and zearalenone (Z)-contaminated diets were fed: .7, 3.1 and 5.8 ppm DON and 0, .05, and .1 ppm Z, respectively. The animals were fed their respective diets for 4 wk followed by the .7:0-ppm diet during wk 5. Feed intake and weight gain varied in a manner reciprocal to the levels of DON-Z in the diets during the first 4 wk (P less than .05). The castrated males had an overall lower weight gain compared with the females receiving the same diet (P less than .05). Gross postmortem changes were not different in either sex and tended to be most prominent in the pigs fed the lower DON:Z-contaminated diets after the first week, although they were seen in pigs fed the higher DON:Z diets after 4 wk of feeding. Lesions included mild to moderated reddening of the fundic mucosa of the stomach, reddening of the mucosa of the small intestine, and mild to moderate enlargement and edema of the mesenteric lymph nodes. Similarly, the severity of histologic changes tended to vary inversely with the concentrations of DON:Z in the diets after the first week but varied with the concentrations of DON:Z after 4 wk. They consisted of vascular congestion with mild to moderate multifocal erosions and degeneration of the mucosa in the stomach and small intestine. Mild to moderate lymphoid degeneration and depletion were also observed in the Peyer's patches of the intestines, bronchial and mesenteric lymph nodes, spleen, tonsil and thymus.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animal Feed; Animals; Body Weight; Dose-Response Relationship, Drug; Feeding Behavior; Female; Gastrointestinal Diseases; Male; Ovariectomy; Resorcinols; Sesquiterpenes; Swine; Swine Diseases; Tissue Distribution; Trichothecenes; Zearalenone

Purified vomitoxin was incorporated into the diet at a level of 20 ppm and fed to male Sprague-Dawley rats ad lib. for 90 days. Few clinical signs of toxicity were observed. Rats in the vomitoxin treatment group were less efficient in converting feed into body mass, but there was no feed refusal. Terminal body weight was reduced in the vomitoxin treatment group. There were no statistically significant effects on serum enzyme levels, haematological parameters or tissue lesions, or on liver detoxification systems, as reflected in levels of microsomal cytochrome P-450 or in glutathione S-transferase activity.

Topics: Animals; Body Weight; Cytochrome P-450 Enzyme System; Glutathione Transferase; Liver; Male; Rats; Rats, Inbred Strains; Sesquiterpenes; Swine; Trichothecenes; Zearalenone

13C NMR spectroscopic investigations on the biosynthesis of mycotoxins produced by Fusarium graminearum (M69) were carried out through the incorporation of [1-13C]- and [2-13C]acetate precursors. The major secondary metabolites produced by this species in still culture were deoxynivalenol (3,7,15-trihydroxy-12,13-epoxytrichothec-9-en-one), 15-acetyldeoxynivalenol, zearalenone, and butenolide. [1-13C]- and [2-13C]acetate were incorporated in alternate carbon atoms in zearalenone, consistent with the head to tail condensation of nine acetate units. The trichothecenes were enriched in a manner consistent with the condensation of three mevalonate units. 13C/13C couplings, observed between C-5 and C-12, as well as between C-6 and C-15 of 15-acetyldeoxynivalenol, confirms the current hypothesis of formation of the trichothecene ring system by cyclization of farnesyl pyrophosphate. The incorporation pattern in ergosterol is also consistent with a mevalonate origin, while the adjacent incorporation of acetate methyl groups in butenolide suggests a glutamate precursor. The degree of enrichment in the secondary metabolites, which ranged from 3 to 10% at each carbon site, was observed in the 13C NMR spectra of the crude fungal extracts to be dependent on the timing of acetate addition to the culture. The specific toxins produced together with the quantity of each, were also found to be dependent on the timing of acetate addition. Competition between the three biosynthetic pathways of secondary metabolism, i.e. polyketide, mevalonate, and amino acid for the labeled acetate in this organism is a complex function of culture conditions.

Topics: Acetates; Fusarium; Magnetic Resonance Spectroscopy; Mycotoxins; Trichothecenes; Zearalenone

Weaner pigs on a farm near Beaudesert in south eastern Queensland refused to eat feed comprised largely of wheat and barley. Older pigs consumed small amounts and some prepubertal gilts subsequently displayed enlarged and reddened vulvas. Wheat, barley and triticale were grown on the farm during 1983, which was unusually and persistently wet. The wheat and triticale were harvested and stored for about 3 weeks with moisture contents above 14% before being fed. Samples of the wheat and triticale contained pale pink grains, which can indicate infection by the fungus Fusarium graminearum Schw. On analysis 2 mycotoxins known to be produced by F. graminearum were detected, deoxynivalenol (vomitoxin) which causes feed refusal and vomiting, and zearalenone which causes oestrogenic effects. Concentrations of deoxynivalenol in the wheat, triticale and barley were 34, 10, and less than 0.1 mg/kg respectively. Concentrations of zearalenone were 6.2, 2.8 and 0.1 mg/kg respectively. Subsequently, F. graminearum was isolated from grains and crop residues. Although the wet weather contributed to F. graminearum infection of the crops before harvest, most of the toxins probably developed during storage.

Topics: Animal Feed; Animals; Australia; Edible Grain; Food Preferences; Sesquiterpenes; Swine; Trichothecenes; Zearalenone

Zearalenone, an estrogenic mycotoxin of Fusarium species, in cereals can be extracted with acetonitrile-water (3:1), purified on a Florisil column, resolved by high-performance liquid chromatography (HPLC) with a Nucleosil 50-10 column using 90% water-saturated chloroform-cyclohexane-acetonitrile-ethanol (50:15:2:1) and quantitated by fluorescence measurement. This method is rapid, simple and reproducible, and detects zearalenone in wheat, barley, corn and other cereals with picogram sensitivity. A combination of this HPLC method with a gas-liquid chromatographic method for trichothecenes may be applied to the simultaneous detection of Fusarium mycotoxins (zearalenone, nivalenol and deoxynivalenol) in cereals.

Topics: Chromatography, High Pressure Liquid; Edible Grain; Food Microbiology; Hordeum; Resorcinols; Spectrometry, Fluorescence; Trichothecenes; Triticum; Zea mays; Zearalenone

Fusarium spp. isolated from plant materials grown in the hot, humid climate of North Carolina were tested for production of mycotoxins. Isolates of F. acuminatum, F. graminearum, F. moniliforme, F. oxysporum, and F. solani produced zearalenone while isolates of F. equiseti and F. graminearum produced T-2 toxin and deoxynivalenol, respectively. This is the first report of zearalenone production by F. solani. The toxins were identified by capillary gas chromatography-mass spectrometry. These findings suggest that there are toxigenic strains of Fusarium indigenous to the warmer regions of the USA and that fasariotoxicoses of animals in this region are not necessarily the result of importing toxic grains from the cooler, upper midwestern USA.

Topics: Animal Feed; Food Contamination; Food Microbiology; Fusarium; Mass Spectrometry; Medicago sativa; North Carolina; Plants; Resorcinols; Sesquiterpenes; T-2 Toxin; Temperature; Trichothecenes; Zearalenone

Thirty-three samples of wheat of the 1982 crop year from Kansas and Nebraska were analyzed for deoxynivalenol, T-2 toxin, zearalenone, and aflatoxin. Deoxynivalenol was identified in 31 of 33 samples, zearalenone was identified in 3 of 33 samples, and aflatoxin B1 was identified in 23 of 31 samples. One 1982 wheat sample from Illinois and one from Texas were also contaminated with deoxynivalenol at 1,200 and 600 ng/g, respectively. None of the samples contained detectable T-2 toxin. The mean concentration of deoxynivalenol was 1,782 +/- 262 ng/g, and the concentrations of aflatoxin B1 ranged from 0.8 to 17.0 ng/g, with a mean of 3.37 +/- 0.7. Zearalenone concentrations of the three positive samples were 35, 90, and 115 ng/g. However, density segregation of two other samples which tested negative yielded light fractions, comprising less than 2% of the samples, contaminated at 230 and 254 ng of zearalenone per g; calculated zearalenone concentrations for these two samples were below the limit of detection of the method. The high frequency of aflatoxin B1 and deoxynivalenol in wheat from the 1982 crop is unprecedented, as is the simultaneous contamination of some samples with deoxynivalenol, zearalenone, and aflatoxin B1.

Topics: Aflatoxins; Food Analysis; Food Contamination; Kansas; Nebraska; Resorcinols; Sesquiterpenes; Trichothecenes; Triticum; Zearalenone

During the 1981 corn harvest season in Illinois and surrounding states, cold wet weather enhanced the growth of Fusarium graminearum, with resulting contamination by vomitoxin and, to a lesser extent, zearalenone. Of 342 feed samples analyzed, 274 contained vomitoxin at a concentration ranging from 0.1 to 41.6 ppm (mean, 3.1 ppm) and 40 samples contained zearalenone at a concentration ranging from 0.1 to 8 ppm (mean, 0.66 ppm). Animal health problems and reduced growth performance were observed mainly in swine fed vomitoxin-contaminated rations. The predominant clinical complaints, in decreasing frequency were: reproductive problems (50%), feed refusal (43%), reduced weight gain (25%), diarrhea (17%), death (14%), and emesis (11%).

Topics: Animal Feed; Animals; Body Weight; Diarrhea; Edible Grain; Feeding Behavior; Female; Food Contamination; Gastroenteritis; Illinois; Indiana; Iowa; Pregnancy; Pregnancy Complications; Sesquiterpenes; Swine; Swine Diseases; Trichothecenes; Weather; Zearalenone

Three methods for detecting toxigenic fusaria in culture were compared by using known producers of zearalenone, zearalenol, T-2 toxin, and deoxynivalenol. Moist, autoclaved rice cultures of known toxigenic isolates grown in 20-ml tubes yielded oily extracts containing compounds which interfered with qualitative and quantitative analysis for the mycotoxins. Vermiculite moistened with nutrient broth in 20-ml tubes yielded a much cleaner extract. Growing the fungi on a liquid medium required a shorter incubation period, but yields of T-2 toxin and deoxynivalenol were low and variable, and the method required greater space in the incubator. Screening of the extracts by thin-layer chromatography with colorimetric spray reagents to detect the presence of these toxins permitted reduction in the number of extracts quantified by the more lengthy gas-liquid chromatographic method. Culturing in nutrient broth on vermiculite in tubes coupled to a qualitative screen before quantitation proved to be a convenient, inexpensive, and relatively rapid method that enabled reliable screening of a large number of Fusarium isolates for toxin production as compared with prior methods.

Topics: Culture Media; Fusarium; Resorcinols; Sesquiterpenes; Species Specificity; Trichothecenes; Zearalenone; Zeranol

Deoxynivalenol (DON), 3-acetyldeoxynivalenol (acetylDON) and zearalenone (Ze) were examined for their in vitro effect on mitogen-induced lymphocyte blastogenesis using rat or human peripheral blood lymphocytes (PBL). A dose-dependent reduction of lymphocyte proliferation was demonstrated for each mycotoxin. However, the inhibitory effect of DON was significantly higher than that of the acetylated compound. Concentrations of 90 ng/ml and 220 ng/ml inhibited rat and human lymphocyte blastogenesis by 50%, respectively, whereas 450 ng/ml and 1060 ng/ml acetylDON were required to produce the same effect. The amount of Ze necessary to inhibit blastogenesis by 50% was 250 times greater than required for DON. There was no evidence of cell death and combinations of DON and Ze did not alter the expected response.

Topics: Animals; Drug Synergism; Humans; Lymphocyte Activation; Phytohemagglutinins; Rats; Resorcinols; Sesquiterpenes; Species Specificity; Thymidine; Trichothecenes; Zearalenone

Three isolates of Fusarium graminearum (DAOM 180377, 180378, and 180379) were screened for their ability to produce mycotoxins on the solid substrates corn and rice. They all produced deoxynivalenol and zearalenone on corn. On rice, only DAOM 180378 and 180379 produced significant amounts of these mycotoxins, with levels of deoxynivalenol being much higher than those of zearalenone. The effects of the initial moisture content before autoclaving, incubation temperature, and time were studied with isolate DAOM 180378. At 19.5 degrees C the main product was zearalenone, whereas at 25 degrees C both deoxynivalenol and zearalenone were formed. Higher incubation temperatures (28 degrees C) favored deoxynivalenol formation, the maximum amount being 515 ppm (515 micrograms/g) formed after 24 days at an initial moisture content of 40%. The maximum level of zearalenone produced at the same temperature was 399 ppm, but at an initial moisture content of 35%. Other factors, such as pH, oxygen and carbon dioxide concentrations, and size of the culture flask also appeared to affect the production of mycotoxins.

Topics: Food Contamination; Food Microbiology; Fusarium; Mycotoxins; Oryza; Resorcinols; Sesquiterpenes; Trichothecenes; Zea mays; Zearalenone

Out of the 25 strains of the genus Fusarium isolated from wheat visibly infested by moulds, three strains of F. graminearum were found to produce vomitoxin and zearalenone; the respective amounts of these toxins per 1 kg of wheat were 70, 100 and 600 mg vomitoxin, and 600, 400 and 250 mg zearalenone. The production of vomitoxin showed a marked decrease when the cultures were re-inoculated. An analytic determination of vomitoxin in cereals was worked out by the method of high-pressure liquid chromatography with a limit sensitivity of 20 micrograms vomitoxin per kg and by the method of thin-layer chromatography with a limit sensitivity of 100 micrograms vomitoxin per kg. The embryotoxicity of vomitoxin measured in chick embryos incubated for 40 hours was in the order of 1 microgram per embryo.

Topics: Animals; Chick Embryo; Fusarium; Sesquiterpenes; Trichothecenes; Triticum; Zearalenone

Corn purposely infected with Fusarium graminearum was found to contain 800 to 900 mg vomitoxin/kg. Contaminated corn was substituted for control corn at 0, 6, 12, 18, and 24% in a corn-soybean meal ration. Broiler cockerels were given each experimental diet from 6 to 11 days of age; then sample groups were necropsied. Remaining birds were subsequently offered commercial starter for 2 days and sample groups again necropsied. Growth and diet consumption were not significantly reduced until contaminated corn exceeded 12% of the ration (116 mg vomitoxin/kg). Alertness, coordination, and feathering appeared normal regardless of treatment. Birds that received contaminated corn exhibited plaques in the mouth and gizzard erosions proportional to the level of substitution. All lesions were generally restricted to the epithelial layer and no liver or kidney involvement could be demonstrated. A short return to uncontaminated feed eliminated most lesions. Fowl appear to be considerably more tolerant of vomitoxin than swine.

Topics: Animals; Body Weight; Chickens; Diet; Drug Tolerance; Food Contamination; Fusarium; Gizzard, Avian; Male; Mouth Mucosa; Poultry Diseases; Sesquiterpenes; Trichothecenes; Zea mays; Zearalenone

A corn culture of Fusarium roseum was added to a standard corn-soybean swine gestation ration. Low, middle, and high dosage mixed feeds contained 7, 38, and 64 mg of zearalenone/kg of feed (7, 38, and 64 ppm) and 0.5, 2.5, and 4.5 mg of deoxynivalenol/kg, respectively. Control feed was the standard ration without added F roseum corn culture. Mature gilts were bred by natural service and fed control or F roseum molded feed from 3 to 34 days after breeding. The main effect of the molded feed was an inhibition of fetal development, with decreased numbers of fetuses present in treated animals at slaughter (38 to 43 days after breeding). Normal litters were present in 7 of 8 control animals, in 2 of 4 gilts given the low-dosage feed, in 1 of 4 gilts given the medium dosage, and in 0 of 4 given the high-dosage feed. Corpora lutea were maintained in all treated animals, as evidenced by serum progesterone concentrations. Serum estradiol concentrations were decreased in gilts in the middle- and high-dosage groups. The genital system of the gilts fed low- and middle-dosage feeds had a gross and microscopic appearance similar to that of the pregnant controls and reflected prolonged progesterone stimulation. Morphologic changes in the genital system of the high-dosage group were intermediate between changes induced by progesterone and those induced by estrogen. Clinical signs of hyperestrogenism and partial feed refusal were noticed in only some of the high-dosage group animals.

Topics: Animals; Corpus Luteum; Embryo Implantation; Endometrium; Female; Fetus; Fusarium; Growth; Litter Size; Mucous Membrane; Pregnancy; Pregnancy, Animal; Resorcinols; Swine; Swine Diseases; Trichothecenes; Zea mays; Zearalenone

Analysis of a 'yellow rain' sample by selected ion monitoring revealed the presence of three trichothecenes: T-2 toxin, diacetoxyscirpenol and 4-deoxynivalenol in concentrations of at least 48, 42 and 58 ppm, respectively. The concentration of zearalenone, another Fusarium mycotoxin, was estimated to be at least 265 ppm. Evidence for a formulation which contained polyethylene glycol was also obtained.

Topics: Chemical Warfare Agents; Environmental Pollutants; Fusarium; Gas Chromatography-Mass Spectrometry; Laos; Mass Spectrometry; Mycotoxins; Rain; T-2 Toxin; Trichothecenes; Weather; Zearalenone

An episode of suboptimal growth, poor feathering and behavioural abnormalities in broilers in Scotland during the winter of 1980-81 is described. This was considered to be associated with mould-contaminated maize and wheat components of the feed, from which fusaria were isolated in persistently high numbers. Four species, Fusarium culmorum, F tricinctum, F nivale and F moniliforme, were identified. Chloroform extracts of the raw materials and of an artificial medium in which three of the Fusarium species were cultured proved toxic to tissue cultures of a human epithelial cell line (HEp II). Specific identification by thin layer chromatography of the mycotoxins deoxynivalenol, zearalenone and diacetoxyscirpenol was achieved in some extracts. In addition, several other areas of the chromatograms were found to be toxic in the HEp II cell system and these may contain toxins for which standards were not available or, alternatively, previously uncharacterised fungal metabolites. It was concluded that the toxins produced by the fusaria were major contributing factors to the disease symptoms shown by the birds.

Topics: Animal Feed; Animals; Chickens; Food Microbiology; Foodborne Diseases; Fusarium; Mycotoxins; Poultry Diseases; Scotland; Species Specificity; Trichothecenes; Zearalenone

Sixteen Fusarium isolates belonging to F. graminearium Schw. and F. culmorum (W.G. Smith) Sacc. produced vomitoxin and zearalenone on cracked corn at 28 degrees C. Quantitation for vomitoxin was by gas-liquid chromatography. This toxin was produced in quantities of 5 to 236 microgram/g of fermented corn. Vomitoxin showed weak antibiotic activity against Penicillium digitatum Sacc., Mucor ramannianus Möller, and Saccharomyces bayanus Sacc., but did not inhibit gram-positive, gram-negative, or acid fast bacteria. The two molds and the yeast were inhibited by T-2 toxin at 5 micrograms, and diacetoxyscirpenol inhibited the molds at 5 micrograms and the yeast at 50 micrograms.

Topics: Bacteria; Fungi; Fusarium; Resorcinols; Sesquiterpenes; Trichothecenes; Zea mays; Zearalenone

The mycotoxins zearalenone (2.8 micrograms/g), deoxynivalenol (1.5 microgram/g), and T-2 toxin (110 ng/g) have been found in the pith of corn stalks standing in the field. Such contaminated stalks may contribute to mycotoxicoses of farm animals.

Topics: Fusarium; Minnesota; Plant Diseases; Resorcinols; Sesquiterpenes; T-2 Toxin; Trichothecenes; Zea mays; Zearalenone

Many cereal grains have been studied for their suitability as substrates for the fermentative production of mycotoxins. However, except for aflatoxin, wild rice has not been investigated. Hence, five mold cultures known to produce the mycotoxins ochratoxin-A, penicillic acid, patulin, vomitoxin, and zearalenone were grown on wild rice under varying conditions of moisture and temperature to determine whether this grain would serve as a suitable substrate for toxin production. Under appropriate fermentation conditions, good yields of ochratoxin-A and moderate amounts of patulin were obtained, but only small amounts of penicillic acid, vomitoxin, and zearalenone were elaborated. An extract from a sample of naturally molded wild rice contained 0.8 microgram of patulin per g of rice. The predominating mold was identified as Aspergillus clavatus. Under identical cultural conditions, this isolate and a known patulin-producing strain of A. clavatus yielded approximately equivalent amounts of the mycotoxin.

Topics: Aspergillus; Culture Media; Fermentation; Fusarium; Ochratoxins; Oryza; Patulin; Penicillic Acid; Penicillium; Sesquiterpenes; Species Specificity; Trichothecenes; Zearalenone