fusarenon-x and nivalenol

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

Trichothecenes (TCT) are very common mycotoxins. While the effects of DON, the most prevalent TCT, have been extensively studied, less is known about the effect of other trichothecenes. DON has ribotoxic, pro-inflammatory, and cytotoxic potential and induces multiple toxic effects in humans and animals. Although DON is not genotoxic by itself, it has recently been shown that this toxin exacerbates the genotoxicity induced by model or bacterial genotoxins. Here, we show that five TCT, namely T-2 toxin (T-2), diacetoxyscirpenol (DAS), nivalenol (NIV), fusarenon-X (FX), and the newly discovered NX toxin, also exacerbate the DNA damage inflicted by various genotoxins. The exacerbation was dose dependent and observed with phleomycin, a model genotoxin, captan, a pesticide with genotoxic potential, and colibactin, a bacterial genotoxin produced by the intestinal microbiota. For this newly described effect, the trichothecenes ranked in the following order: T-2>DAS > FX > NIV ≥ DON ≥ NX. The genotoxic exacerbating effect of TCT correlated with their ribotoxic potential, as measured by the inhibition of protein synthesis. In conclusion, our data demonstrate that TCT, which are not genotoxic by themselves, exacerbate DNA damage induced by various genotoxins. Therefore, foodborne TCT could enhance the carcinogenic potential of genotoxins present in the diet or produced by intestinal bacteria.

Topics: Animals; DNA Damage; Humans; Mutagens; Trichothecenes

Topics: Chromatography, Liquid; Mycotoxins; Reproducibility of Results; Tandem Mass Spectrometry; Trichothecenes

Type B trichothecenes commonly contaminate cereal grains and include five structurally related congeners: deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), fusarenon X (FX), and nivalenol (NIV). These toxins are known to have negative effects on human and animal health, particularly affecting food intake. However, the pathophysiological basis for anorexic effect is not fully clarified. The purpose of this study is to explore the potential roles of the brain-gut peptides substance P (SP) and glucagon-like peptide-1

Topics: Amides; Animals; Anorexia; Appetite Depressants; Glucagon-Like Peptide 1; Humans; Substance P; Trichothecenes; Trichothecenes, Type B

Exposure to environmental contaminants might play an important role in neurodegenerative disease pathogenesis, such as Parkinson´s disease (PD) and Alzheimer´s disease (AD). For the first time in Spain, the plasmatic levels of 19 mycotoxins from patients diagnosed with a neurodegenerative disease (44 PD and 24 AD) and from their healthy companions (25) from La Rioja region were analyzed. The studied mycotoxins were aflatoxins B1, B2, G1, G2 and M1, T-2 and HT-2, ochratoxins A (OTA) and B (OTB), zearalenone, sterigmatocystin (STER), nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, deepoxy-deoxynivalenol, neosolaniol, diacetoxyscirpenol and fusarenon-X. Samples were analyzed by LC-MS/MS before and after treatment with β-glucuronidase/arylsulfatase in order to detect potential metabolites. Only OTA, OTB and STER were detected in the samples. OTA was present before (77% of the samples) and after (89%) the enzymatic treatment, while OTB was only detectable before (13%). Statistically significant differences in OTA between healthy companions and patients were observed but the observed differences might seem more related to gender (OTA levels higher in men,

Topics: Alzheimer Disease; Biological Monitoring; Chromatography, Liquid; Humans; Mycotoxins; Neurodegenerative Diseases; Ochratoxins; Parkinson Disease; Sterigmatocystin; Tandem Mass Spectrometry; Trichothecenes; Zearalenone

A liquid chromatography-tandem mass spectrometry method was developed for simultaneous determination of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3Ac-DON), 15-acetyldeoxynivalenol (15Ac-DON), DON-3-glucoside (DON-3Glc) nivalenol and fusarenone-X in feedstuffs. Different techniques of sample preparation were tested: solid-liquid-extraction, QuEChERS, solid phase extraction with OASIS HLB columns or immunoaffinity columns and a Mycosep 225 Trich column. None of the six immunoaffinity columns tested showed cross-reactivity to all of the mycotoxins. Surprisingly, the results show that if the immunoaffinity columns bound 3Ac-DON, then they did not bind 15Ac-DON. The most efficient sample preparation was achieved with a Mycosep 225 Trich column clean-up. The chromatography was optimised to obtain full separation of all analytes (including 3Ac-DON and 15Ac-DON isomeric form). The validation results show the relative standard deviations for repeatability and reproducibility varied from 4% to 24%. The apparent recovery ranged between 92% and 97%, and the limit of quantification described a 1.30 to 50 µg/kg range. The method trueness was satisfactory, as assessed by a proficiency test and analysis of reference material. A total of 99 feed samples were analysed by the developed method, revealing the presence of DON and DON-3Glc in 85% and 86% of examined animal feeds, respectively at concentrations between 1.70 and 1709 µg/kg. The ratios DON-3Glc to DON in the surveyed feedstuffs were from a low of 3% to high of 59%.

Topics: Animal Feed; Chromatography, Liquid; Food Microbiology; Fungi; Limit of Detection; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry; Trichothecenes

Topics: Apoptosis; Biological Assay; Cell Survival; Dose-Response Relationship, Drug; Drug Synergism; Flow Cytometry; Humans; Inhibitory Concentration 50; Jurkat Cells; Trichothecenes

Lymphocytes are involved in the adaptive immune response and are highly sensitive to type B trichothecenes. In grains and their products, deoxynivalenol (DON) is the most widely distributed trichothecene. It usually co-occurs with other type B members, such as nivalenol (NIV) and fusarenon-X (FX), because they are all produced by the same Fusarium fungi. However, the combined effects of mycotoxins are complex and cannot be predicted based on individual toxicity. Thus, the adverse effects of combined toxins are of increasing concern. The aim of this study was to compare the toxicity to lymphoid tissues of mice of DON alone or mixed with NIV or FX. Forty, 3-week-old male ICR mice were given a single oral administration of a vehicle control, one toxin, binary, or ternary mixtures and then sacrificed at 12 h after exposure. Mice treated with FX alone showed marked nuclear condensation and fragmentation of lymphocytes in the cortical thymus and germinal center of Peyer's patches and spleen. Similarly, these animals clearly displayed TUNEL- and Caspase-3-positive cells in the regions. In contrast, minimal changes were noticed in the lymphoid tissues of mice receiving combined toxins when compared to this toxin alone. In addition, oral exposure to FX alone significantly up-regulated the relative expression of Bax, Caspase-3, Caspase-9, and Trp53. These data increase our understanding of the toxic actions of DON, NIV, and FX alone or in combination to lymphocytes and can be used to assess the possible risk associated with their co-occurrences in foodstuffs to human and animal health.

Topics: Administration, Oral; Animals; Apoptosis; In Situ Nick-End Labeling; Lymphocytes; Male; Mice; Mice, Inbred ICR; Mycotoxins; Trichothecenes

Topics: Animals; Feces; Goats; Male; Microsomes, Liver; Toxicokinetics; Trichothecenes

Topics: Cell Line; Cell Survival; Epithelial Cells; Humans; Interleukin-6; Interleukin-8; Respiratory System; Trichothecenes

Deoxynivalenol is a food borne mycotoxin belonging to the trichothecenes family that may cause severe injuries in human and animals. The inhibition of protein synthesis via the interaction with the ribosome has been identified as a crucial mechanism underlying toxic action. However, it is not still fully understood how and to what extent compounds belonging to trichothecenes family affect human and animal health. In turn, this scenario causes delay in managing the related health risk. Aimed at supporting the hazard identification process, the in silico analysis may be a straightforward tool to investigate the structure-activity relationship of trichothecenes, finding out molecules of possible concern to carry forth in the risk assessment process. In this framework, this work investigated through a molecular modeling approach the structural basis underlying the interaction with the ribosome under a structure-activity relationship perspective. To identify further forms possibly involved in the total trichothecenes-dependent ribotoxic load, the model was challenged with a set of 16 trichothecene modified forms found in plants, fungi and animals, including also compounds never tested before for the capability to bind and inhibit the ribosome. Among them, only the regiospecific glycosylation in the position 3 of the sesquiterpenoid scaffold (i.e. T-2 toxin-3-glucuronide, α and β isomers of T-2 toxin-3-glucoside and deoxynivalenol-3-glucuronide) was found impairing the interaction with the ribosome, while the other compounds tested (i.e. neosolaniol, nivalenol, fusarenon-X, diacetoxyscirpenol, NT-1 toxin, HT-2 toxin, 19- and 20-hydroxy-T-2 toxin, T-2 toxin triol and tetraol, and 15-deacetyl-T-2 toxin), were found potentially able to inhibit the ribosome. Accordingly, they should be included with high priority in further risk assessment studies in order to better characterize the trichothecenes-related hazard.

Topics: DNA Mismatch Repair; Food Contamination; Food Microbiology; Glucuronides; Mycotoxins; Ribosomes; Structure-Activity Relationship; T-2 Toxin; Trichothecenes

In case of mycotoxin contaminations, food and feedstuff are usually contaminated by more than one toxin. However toxicological data concerning the effects of mycotoxin combinations are sparse. The intestinal epithelium is the first barrier against food contaminants and this constantly renewing organ is particularly sensitive to mycotoxins. The aim of this study was to investigate the effects of deoxynivalenol (DON) and four other type B trichothecenes (TCTB), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), nivalenol (NIV) and fusarenon-X (FX) alone or in combination on intestinal epithelial cells. Proliferating, non-transformed IPEC-1 cells were exposed to increasing doses of TCTB, alone or in binary mixtures and mycotoxin-induced cytotoxicity was measured with MTT test. The toxicological interactions were assessed using the isobologram-Combination index method. The five tested mycotoxins and their mixtures had a dose-dependent effect on the proliferating enterocytes. DON-NIV, DON-15-ADON and 15-ADON-3-ADON combinations were synergistic, with magnitude of synergy for 10 % cytotoxicity ranging from 2 to 7. The association between DON and 3-ADON also demonstrated a synergy but only at high doses, at lower doses antagonism was noted. Additivity was observed between NIV and FX, and antagonism between DON and FX. These results indicate that the simultaneous presence of mycotoxins in food commodities and diet may be more toxic than predicted from the mycotoxins alone. This synergy should be taken into account considering the frequent co-occurrence of TCTB in the diet.

Topics: Animals; Cell Culture Techniques; Cell Survival; Cells, Cultured; Dose-Response Relationship, Drug; Drug Synergism; Epithelial Cells; Intestinal Mucosa; Swine; Trichothecenes

Cereal grain contamination by trichothecene mycotoxins is known to negatively impact human and animal health with adverse effects on food intake and growth being of particular concern. The head blight fungus Fusarium graminearum elaborates five closely related 8-ketotrichothecene congeners: (1) deoxynivalenol (DON), (2) 3-acetyldeoxynivalenol (3-ADON), (3) 15-acetyldeoxynivalenol (15-ADON), (4) fusarenon X (FX), and (5) nivalenol (NIV). While anorexia induction in mice exposed intraperitoneally to DON has been linked to plasma elevation of the satiety hormones cholecystokinin (CCK) and peptide YY₃₋₃₆ (PYY₃₋₃₆), the effects of oral gavage of DON or of other 8-keotrichothecenes on release of these gut peptides have not been established. The purpose of this study was to (1) compare the anorectic responses to the aforementioned 8-ketotrichothecenes following oral gavage at a common dose (2.5 mg/kg bw) and (2) relate these effects to changes plasma CCK and PYY₃₋₃₆ concentrations. Elevation of plasma CCK markedly corresponded to anorexia induction by DON and all other 8-ketotrichothecenes tested. Furthermore, the CCK1 receptor antagonist SR 27897 and the CCK2 receptor antagonist L-365,260 dose-dependently attenuated both CCK- and DON-induced anorexia, which was consistent with this gut satiety hormone being an important mediator of 8-ketotrichothecene-induced food refusal. In contrast to CCK, PYY₃₋₃₆ was moderately elevated by oral gavage with DON and NIV but not by 3-ADON, 15-ADON, or FX. Taken together, the results suggest that CCK plays a major role in anorexia induction following oral exposure to 8-ketotrichothecenes, whereas PYY₃₋₃₆ might play a lesser, congener-dependent role in this response.

Topics: Administration, Oral; Animals; Anorexia; Chemokines, CC; Cholecystokinin; Female; Mice; Mycotoxins; Peptide Fragments; Peptide YY; Receptor, Cholecystokinin B; Receptors, Cholecystokinin; Trichothecenes

Fusarenon-X (FX) is one of the trichothecene mycotoxins mainly produced by Fusarium crookwellense, which naturally occurs in agricultural commodities such as wheat and barley. To investigate the toxicokinetics of FX and its metabolite nivalenol (NIV), FX was then administered intravenously or orally to piglets at a dosage of 1mg/kg body weight. The concentrations of FX and NIV in the plasma and various tissues were measured using LC-MS/MS. The plasma concentrations of FX in the piglets were determined up to 24h and 48h after iv and po administration, respectively, and the concentration of NIV was detected up to 12h after both types of administration. The Cp(0) of FX was 580.28 ± 140.81 ng/ml after iv administration. The values of t1/2β, Vss and Foral were 1.71 ± 0.74 h, 0.009 ± 0.002 ml and 74.40 ± 18.96%, respectively. FX and NIV were detectable in the vital organs up to 24h after po administration. The peak level of FX in the liver, the kidney, and the spleen, respectively, were 165.95 ± 9.68 ng/g, 66.29 ± 8.48 and 7.35 ± 0.69 ng/g at 3h following po administration. In vitro of liver postmitochondrial fractions with FX demonstrated that the liver and kidney are capable of FX-to-NIV metabolism.

Topics: Animals; Chromatography, Liquid; Female; Swine; Tandem Mass Spectrometry; Trichothecenes

Deoxynivalenol (DON) is the most prevalent trichothecene mycotoxin in crops in Europe and North America. DON is often present with other type B trichothecenes such as 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), nivalenol (NIV) and fusarenon-X (FX). Although the cytotoxicity of individual mycotoxins has been widely studied, data on the toxicity of mycotoxin mixtures are limited. The aim of this study was to assess interactions caused by co-exposure to Type B trichothecenes on intestinal epithelial cells. Proliferating Caco-2 cells were exposed to increasing doses of Type B trichothecenes, alone or in binary or ternary mixtures. The MTT test and neutral red uptake, respectively linked to mitochondrial and lysosomal functions, were used to measure intestinal epithelial cytotoxicity. The five tested mycotoxins had a dose-dependent effect on proliferating enterocytes and could be classified in increasing order of toxicity: 3-ADON<15-ADON≈DON

Topics: Algorithms; Caco-2 Cells; Cell Survival; Coloring Agents; Dose-Response Relationship, Drug; Drug Synergism; Epithelial Cells; Humans; Intestinal Mucosa; Mycotoxins; Tetrazolium Salts; Thiazoles; Trichothecenes

Although the acute toxic effects of trichothecene mycotoxin deoxynivalenol (DON or vomitoxin), a known cause of human food poisoning, have been well characterized in several animal species, much less is known about closely related 8-ketotrichothecenes that similarly occur in cereal grains colonized by toxigenic fusaria. To address this, we compared potencies of DON, 15-acetyldeoxynivalenol (15-ADON), 3-acetyldeoxynivalenol (3-ADON), fusarenon X (FX), and nivalenol (NIV) in the mink emesis model following intraperitoneal (ip) and oral administration. All five congeners dose-dependently induced emesis by both administration methods. With increasing doses, there were marked decreases in latency to emesis with corresponding increases in emesis duration and number of emetic events. The effective doses resulting in emetic events in 50% of the animals for ip exposure to DON, 15-ADON, 3-ADON, FX, and NIV were 80, 170, 180, 70, and 60 µg/kg bw, respectively, and for oral exposure, they were 30, 40, 290, 30, and 250 µg/kg bw, respectively. The emetic potency of DON determined here was comparable to that reported in analogous studies conducted in pigs and dogs, suggesting that the mink is a suitable small animal model for investigating acute trichothecene toxicity. The use of a mouse pica model, based on the consumption of kaolin, was also evaluated as a possible surrogate for studying emesis but was found unsuitable. From a public health perspective, comparative emetic potency data derived from small animal models such as the mink should be useful for establishing toxic equivalency factors for DON and other trichothecenes.

Topics: Administration, Oral; Animals; Dose-Response Relationship, Drug; Female; Injections, Intraperitoneal; Male; Mice; Mink; Mycotoxins; Pica; Toxicity Tests; Trichothecenes; Vomiting

While induction of food refusal by the trichothecene mycotoxin deoxynivalenol (DON) has been described in several animal models, much less is known about the anorectic effects of structurally related 8-ketotrichothecenes, 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), fusarenon X (FX) and nivalenol (NIV). Here, we compared the capacities of these congeners to induce anorexia in the mouse. As previously observed for DON, anorectic responses to 3-ADON and 15-ADON in the B6C3F1 female mouse following both intraperitoneal (IP) and oral exposure were transient, lasting only a few hours, with food intake recovering to control levels within 16 h. For both ADONs, the no observed adverse effect levels (NOAEL) and lowest observed adverse effect levels (LOAEL) were 0.5 and 1mg/kg bw following IP exposure, respectively, and 1 and 2.5mg/kg bw after oral exposure, respectively. In contrast, food refusal persisted from 48 to 96 h following IP and oral exposure to FX and NIV. For both IP and oral FX exposure, the NOAEL was 0.025 mg/kg bw and LOAEL was 0.25mg/kg bw, whereas the NOAELs and LOAELs for NIV were 0.01 and 0.1mg/kg bw, respectively, after IP exposure and 0.1 and 1mg/kg bw, respectively, following oral exposure. Both these data and a prior DON study suggest that anorectic responses to 8-ketotrichothecenes were always greater when administered IP as compared to oral exposure and follow an approximate rank order of NIV>FX>DON≈3-ADON≈15-ADON for IP exposure and FX>NIV>DON≈3-ADON≈15-ADON for oral exposure. Toxic potency data such as is described here will be applicable to future comparative risk assessments for this important group of trichothecene mycotoxins.

Topics: Animals; Appetite Depressants; Dose-Response Relationship, Drug; Eating; Female; Fusarium; Mice; Mice, Inbred Strains; Mycotoxins; No-Observed-Adverse-Effect Level; Trichothecenes

The objective of the present study was to monitor the occurrence and distribution of a spectrum of trichothecene toxins in different parts of maize plants. Therefore maize plants were sampled randomly from 13 fields in southwest Germany and the fractions kernels, cobs, husks, stalks, leaves and rudimentary ears were analyzed for eight A-type and five B-type trichothecenes. Each of the toxins was found in at least three of the total of 78 samples. The study revealed that both A-type and B-type trichothecenes may be present in all parts of the maize plant but may be unevenly distributed. For the contents of deoxynivalenol, 3- and 15-acetyldeoxynivalenol, nivalenol, scirpentriol, 15-monoacetoxyscirpenol, HT-2 and T-2 toxin significant differences (p < 0.05) were found between different parts of the maize plants whereas no significant differences were observed for fusarenon-X, 4,15-diacetoxyscirpenol, neosolaniol, T-2 triol and T-2 tetraol. Up to twelve toxins co-occurring in one sample were detected. As a group B-type trichothecenes dominated over A-type trichothecenes concerning incidences and levels. Contamination was strongest with rudimentary ears based on incidence and mean and maximum contents; mean contents with few exceptions tended towards a higher level than in other fractions with significant (p < 0.05) differences compared to leaves for seven toxins.

Topics: Food Contamination; Food Microbiology; Germany; T-2 Toxin; Trichothecenes; Zea mays

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

A speedy and selective ultra-HPLC-MS/MS method for simultaneous determination of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-ADON, nivalenol and fusarenon X in traditional Chinese medicines (TCMs) was developed. The method was based on one-step sample cleanup using reliable homemade cleanup cartridges. A linear gradient mobile-phase system, consisting of water containing 0.2% aqueous ammonia and acetonitrile/methanol (90:10, v/v) at a flow rate of 0.4 mL/min, and an Acquity UPLC HSS T3 column (100 mm x 2.1 mm, 1.8 microm) were employed to obtain the best resolution of the target analytes. [(13)C(15)]-DON was used as the internal standard to accomplish as accurate as possible quantitation. The established method was further validated by determining the linearity (R(2) > or = 0.9990), sensitivity (LOQ, 0.29-0.99 microg/kg), recovery (88.5-119.5%) and precision (RSD < or = 15.8%). It was shown to be a suitable method for simultaneous determination of DON, 3-ADON, 15-ADON, nivalenol and fusarenon X in various TCM matrices. The utility and practical impact of the method was demonstrated using different TCM samples.

Topics: Chromatography, High Pressure Liquid; Drugs, Chinese Herbal; Medicine, Chinese Traditional; Molecular Conformation; Stereoisomerism; Tandem Mass Spectrometry; Trichothecenes

In order to investigate the comparative fates and dispositions of fusarenon-X (FX) in broilers and ducks, FX was administered i.v. or orally (p.o.) to broilers and ducks. The FX and its metabolite (nivalenol, NIV) were determined in plasma and excreta using gas chromatography-mass spectrometry. The plasma concentrations of FX were determined up to 180 and 120 min in broilers and ducks, respectively, after i.v. and p.o. administration. The NIV was eliminated more slowly than its parent compound. The FX disposition fit an open 2-compartment pharmacokinetic model in broilers and ducks. The elimination half-life (t(1/2beta)) of FX was longer in ducks than in broilers. The elimination rate constant (kel) was higher in broilers than in ducks, whereas the oral bioavailability of FX was higher in ducks than in broilers. The gas chromatography-mass spectrometry profile in plasma showed that a large proportion of FX was recovered as NIV after administration of FX in both broilers and ducks. In vitro incubation of liver microsomal and cytosolic fractions with FX demonstrated that the liver and kidney are capable of the FX-to-NIV conversion. Thus, this study demonstrated that FX is absorbed more efficiently in ducks than in broilers, whereas it is eliminated more slowly in ducks than in broiler chickens. Consequently, the toxicity would have more serious consequences in ducks rather than broilers.

Topics: Animals; Area Under Curve; Biological Availability; Chickens; Ducks; Feces; Female; Half-Life; Male; Trichothecenes

This study aims to assess the genotoxic potential of nivalenol (NIV) and fusarenon X (FusX), produced by various Fusarium on cereals. Toxins were applied in time and dose-dependent experiments to the human enterocyte-like Caco-2 cell-line, both in dividing (undifferentiated) and in 10-12 days post-confluent cells (differentiated). Genotoxicity was evaluated through the alkaline Comet assay in a concentration range defined for each toxin as below the cytotoxicity threshold IC(10), determined by the MTS and the neutral red assays, to prevent false positive results because of DNA damage stemming from necrosis. Thus, genotoxicity was explored in the sub-cytotoxic 0-0.5 microM and 0-0.05 microM ranges respectively for NIV and FusX as the latter was found about 10-fold more cytotoxic than NIV. For both toxins, a 3h exposure did not cause any DNA damage, unlike after 24 and 72 h exposure in post confluent Caco-2 cells where DNA damage was significantly observed with a dose-dependent relationship. In dividing cells, only FusX increases DNA strand breaks in the 0.01-0.05 microM range after 72 h. These results demonstrated the existence of a genotoxic potential for NIV and FusX at low exposure levels and could contribute to the risk assessment process of these toxins that are of growing concern.

Topics: Caco-2 Cells; Cell Line, Tumor; Cell Survival; Comet Assay; DNA Damage; Dose-Response Relationship, Drug; Enterocytes; Fusarium; Humans; Mutagens; Mycotoxins; Neutral Red; Tetrazolium Salts; Time Factors; Trichothecenes

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

Nivalenol is a mycotoxin produced by certain fungi that are pathogenic to important cereal crops, in particular maize, wheat, and barley. This toxin, 3alpha,4beta,7alpha,15-tetrahydroxy-12,13-epoxytrichothec-9-en-8-one, is found worldwide and is closely related to 4-deoxynivalenol (DON or vomitoxin), a mycotoxin associated with outbreaks of Fusarium head blight in North America. The literature on the toxicity of nivalenol suggests it is similar, if not more toxic, than DON. Despite the development of rapid immunologically based assays for detecting DON, such assays have not existed for detecting nivalenol without chemical modification of the analyte. This paper describes the development of a monoclonal antibody using a nivalenol-glycine protein conjugate. The monoclonal antibody was most specific for an acetylated form of DON (3-Ac-DON), but it exhibited sensitivity and cross-reactivity that were useful for detecting nivalenol and DON at relevant levels without the need to modify either toxin chemically. In an competitive indirect ELISA format, the concentrations of toxins able to inhibit colour development by 50% (IC50) were 1.7, 15.8, 27.5, 68.9, and 1740 ng ml(-1) for the mycotoxins 3-Ac-DON, DON, nivalenol, 15-Ac-DON, and fusarenon-X, respectively. The antibody was also used to develop a competitive direct ELISA for DON and nivalenol, with IC50's of 16.5 ng ml(-1) (DON) and 33.4 ng ml(-1) (nivalenol). These assays are capable of detecting both DON and nivalenol simultaneously, a property that may be useful in regions where these toxins co-occur or in formats, such as immunoaffinity columns, where co-isolation of both toxins is desirable.

Topics: Animals; Antibodies, Monoclonal; Antibody Affinity; Antibody Specificity; Cross Reactions; Enzyme-Linked Immunosorbent Assay; Female; Glycine; Immunotoxins; Mice; Mice, Inbred BALB C; Mycotoxins; Solvents; Trichothecenes

A new, rapid and sensitive method is reported for the multiresidual determination of type A (diacetoxyscirpenol, HT-2 toxin, T-2 toxin) and type B (nivalenol, deoxynivalenol, fusarenon X, 15-O-acetyl-4-deoxynivalenol) trichothecenes in wheat flour samples. Sample extraction was performed with acetonitrile/water mixtures. Mycosep columns were used for a fast and effective clean-up procedure. The analytes were separated by HPLC with a RP C18 column by means of a gradient elution and detected in an ESI-interfaced single quadrupole mass spectrometer. Type B and type A trichothecenes were monitored in the negative and in the positive ion mode, respectively. The method performance is reported in terms of linearity (r2 = 0.999), specificity, accuracy (recoveries from 70-120%) and precision (CV% = 5), the LOQs are in the range 10-20 microg/Kg.

Topics: Chromatography, High Pressure Liquid; Drug Residues; Flour; Food Contamination; Mycotoxins; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; T-2 Toxin; Trichothecenes; Triticum

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

In order to investigate the transfer of nivalenol (NIV) and fusarenon-X (FX) from pregnant to fetal mice and from lactating to suckling mice, (3)H-NIV or (3)H-FX was given p.o. to pregnant or lactating mice. Radioactivity was detected in the whole fetal tissues as well as the fetal liver and kidney, the levels being comparable to those of the maternal tissues. Radioactivity was also detected in the milk, and liver and kidney tissues taken from suckling mice of both (3)H-NIV or (3)H-FX administered dams. HPLC analysis of fetal tissue homogenates from non-labeled FX- or NIV-administered pregnant mice revealed transmission of NIV to fetuses after administration of either toxin. In mice given the non-labeled FX, major and minor peaks of NIV and FX on HPLC were noted in suckling pup tissue homogenates. The results demonstrate that NIV transfers in unchanged form to fetal or suckling mice via placenta or milk, respectively, and that FX does so mainly after being metabolized to NIV in maternal body.

Topics: Animals; Chromatography, High Pressure Liquid; Female; Maternal-Fetal Exchange; Mice; Milk; Mycotoxins; Placenta; Pregnancy; Tissue Distribution; Trichothecenes; Tritium

In 1998, a typhoon struck before rice harvesting in Japan, and the unpolished rice was found to be stained brown. Samples were collected and analyzed for the presence of trichothecenes using GC/MS. The mycotoxins deoxynivalenol (DON), fusarenon-X (Fus.-X) and nivalenol (NIV) were detected and confirmed with GC/MS. The quantity of trichothecenes was determined using GC-ECD. To our knowledge, this is the first report of the presence of the trichothecene Fus.-X in rice.

Topics: Gas Chromatography-Mass Spectrometry; Japan; Oryza; Trichothecenes

In order to investigate the comparative fates of nivalenol (NIV) and 4-acetyl derivative of NIV (fusarenon-X, FX) in mice, 3H-FX or 3H-NIV was given p.o. to mice. Radioactivity was excreted mainly via the urine in mice given 3H-FX, but mainly via the feces in mice given 3H-NIV. The plasma radioactivity reached a peak at 30 or 60 min after the administration of 3H-FX or 3H-NIV, respectively. The plasma peak level was 5 times higher, and the area under curve (AUC) was 10 times higher, in 3H-FX-administered than 3H-NIV-administered mice. These findings clearly demonstrate that FX is absorbed from the gastrointestinal tract more rapidly and efficiently than NIV. The HPLC profile of radioactivity of acetonitrile extracts of urine and feces indicated that FX is rapidly metabolized to NIV after being absorbed from the gastrointestinal tract. In vitro incubation of tissue homogenates with 3H-FX demonstrated that the liver and kidney are the organs responsible for the FX-to-NIV conversion. Thus this study demonstrated that the higher oral toxicity of FX than NIV that has been observed in mice and rats is due to the efficient absorption of FX than NIV from the gastrointestinal tract, followed by its rapid conversion to NIV by the liver and kidney.

Topics: Animals; Chromatography, High Pressure Liquid; Feces; Female; Kidney; Liver; Mice; Mice, Inbred ICR; Molecular Structure; Mycotoxins; Tissue Distribution; Trichothecenes; Tritium

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

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 effect of trichothecene mycotoxins, deoxynivalenol (DON), fusarenon-X (FX) and nivalenol (NIV), on plaque formation of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) in HEp-2 cells was examined. The 50% effective concentrations (EC50) of DON, FX, and NIV for HSV-1 plaque formation were 160, 56, and 120 ng/ml, respectively. Those for HSV-2 plaque formation were 94, 26, and 50 ng/ml, respectively. These three mycotoxins showed about 2-fold higher selectivity to HSV-2 than to HSV-1. Plaque formation of HSV-1 was not inhibited with trichothecenes at concentrations completely inhibiting plaque formation when cells were treated during virus adsorption period or 15 hr before infection. These results indicate that trichothecenes affect replication of HSV-1 after virus adsorption, but not before or during virus adsorption to the host cells.

Topics: Cell Line; Herpesvirus 1, Human; Herpesvirus 2, Human; Humans; Mycotoxins; Trichothecenes; Viral Plaque Assay; Virus Replication

Fusarium crookwellense KF748 (NRRL A-28100) (isolated from dry rotted potato tubers in Central Poland) produced six mycotoxins on both rice and corn substrates at 25 degrees C. The metabolites detected were zearalenone, alpha-trans-zearalenol, beta-trans-zearalenol, fusarin C, and the trichothecenes fusarenone X and nivalenol. This is the first report of formation of alpha-trans-zearalenol, beta-trans-zearalenol, fusarenone X, and nivalenol by F. crookwellense.

Topics: Chromatography, Thin Layer; Fermentation; Fusarium; Mutagens; Mycotoxins; Oryza; Polyenes; Solanum tuberosum; Trichothecenes; Zea mays; Zearalenone; Zeranol

The mycotoxins nivalenol, fusarenon-X, T-2 toxin and zearalenone were checked for clastogenic damage, induction of SCEs and cell cycle delay in Chinese hamster V79-E cells in vitro. The trichothecenes nivalenol, fusarenon-X and T-2 toxin provoked a marked toxicity as especially expressed by cell cycle delay. As compared to toxicity, they are weak clastogens. Addition of S9 mix toxified nivalenol, detoxified T-2 toxin and had no marked influence on fusarenon-X activity. SCE values were slightly increased. It is suggested that the effects observed are unspecific and are caused by inhibition of protein synthesis. Zearalenone was inactive in the three assays. Problems of genotoxicity and carcinogenicity of Fusarium toxins are discussed.

Topics: Animals; Cell Cycle; Cell Line; Chromosome Aberrations; Cricetinae; Cricetulus; Dose-Response Relationship, Drug; Lung; Resorcinols; Sesquiterpenes; Sister Chromatid Exchange; T-2 Toxin; Trichothecenes; Zearalenone