aflatoxin-m1 and Body-Weight

Aflatoxin M1 (AFM1), the only toxin with maximum residue levels in milk, has adverse effects on the intestinal barrier, resulting in intestinal inflammatory disease. Lactoferrin (LF), one of the important bioactive proteins in milk, performs multiple biological functions, but knowledge of the protective effects of LF on the compromised intestinal barrier induced by AFM1 has not been investigated. In the present study, results using Balb/C mice and differentiated Caco-2 cells showed that LF intervention decreased AFM1-induced increased intestinal permeability, improved the protein expression of claudin-3, occludin and ZO-1, and repaired the injured intestinal barrier. The transcriptome and proteome were used to clarify the underlying mechanisms. It was found that LF reduced the intestinal barrier dysfunction caused by AFM1 and was associated with intestinal cell survival related pathways, such as cell cycle, apoptosis and MAPK signaling pathway and intestinal integrity related pathways including endocytosis, tight junction, adherens junction and gap junction. The cross-omics analysis suggested that insulin receptor (INSR), cytoplasmic FMR1 interacting protein 2 (CYFIP2), dedicator of cytokinesis 1 (DOCK1) and ribonucleotide reductase regulatory subunit M2 (RRM2) were the potential key regulators as LF repaired the compromised intestinal barrier. These findings indicated that LF may be an alternative treatment for the compromised intestinal barrier induced by AFM1.

Topics: Aflatoxin M1; Animals; Body Weight; Caco-2 Cells; Cell Membrane Permeability; Claudin-3; Gene Expression Profiling; Gene Expression Regulation; Humans; Intestines; Lactoferrin; Male; Mice; Mice, Inbred BALB C; Occludin; Proteome; RNA, Messenger; Transcriptome; Zonula Occludens-1 Protein

Aflatoxin M1 (AFM1) and ochratoxin A (OTA) are pernicious mycotoxins widely co-existing in the environment. However, nephrotoxicity and underlying mechanism induced by AFM1 coupled with OTA still remain to be explored. In this study, CD-1 mice were treated with 3.5 mg/kg b.w. AFM1, OTA, and AFM1 + OTA for 35 days, and UPLC-MS-based metabolomics method was effectuated to investigate metabolomic profiles of mice kidney. Subsequent experiments on human renal proximal tubular (HK-2) cells were performed to dig out the causal connections between distinguished differential metabolites and nephrotoxicity. Compared with DMSO vehicle group, all three toxin treatments (AFM1 and OTA alone, and in combination) significantly reduced final body weight, and remarkably elevated the concentration of serum creatinine (SCr) and caused abnormal histological phenotypes (shown by histopathological slices). OTA, AFM1 + OTA but not AFM1 reduced the relative weight index of kidney. These phenotypic results indicated that AFM1 and OTA were both toxic to the body, and it seemed that OTA exhibited a notable impairment to kidney while AFM1 had similar but limited effect compared with OTA. Further metabolomics analysis showed that when AFM1 and OTA were combined together, OTA exerted dominant effect on the alteration of metabolic processes. There were few differences in the number of changed metabolites between OTA and AFM1 + OTA group. Among the differentially expressed metabolites affected by OTA, and AFM1 + OTA, lysophosphatidylcholines (LysoPCs) were identified as the main type with significant upregulation, in which LysoPC (16:0) accounted for the most prime proportion. Western blotting results of HK-2 cells showed that single OTA and AFM1 + OTA increased the apoptotic protein expressions of Bax, caspase 3 and PARP, and decreased the expression of Bcl-2; while AFM1 only raised the expression of caspase 3. LysoPC (16:0) but not LysoPC (18:1) lifted the protein level of caspase 3 and PARP in HK-2 cells, and reduced the level of Bcl-2. Taken together, this study is the first effort trying to assess nephrotoxicity of AFM1 with OTA, and we guessed that OTA had a more pronounced toxicity to kidney in contrast to AFM1. No obvious synergism between AFM1 and OTA was found to contribute to the occurrence or development of nephropathy. LysoPC (16:0) might be the pivotal metabolite in response to single OTA and combined AFM1 + OTA engendering renal injury.

Topics: Aflatoxin M1; Animals; Apoptosis; Body Weight; Caco-2 Cells; Carcinogens; Caspase 3; Cell Line; Humans; Kidney; Kidney Diseases; Lysophosphatidylcholines; Male; Metabolomics; Mice; Ochratoxins; Poly (ADP-Ribose) Polymerase-1; Proteomics

This study was conducted to investigate the occurrence of aflatoxin M1 (AFM1) in milk products in China using the competitive enzyme-linked immunosorbent assay method and to estimate the dietary exposure to this toxin through a probabilistic approach. Based on the exposure assessment results, a quantitative cancer potency formula developed by the Joint Food and Agriculture Organization and World Health Organization Expert Committee on Food Additives was applied to assess the cancer risk. AFM1 was detected in 48.07% of the milk samples and 4.49% of the yoghurt samples. No samples contained AFM1 above the current regulatory limit in China. The simulated AFM1 intake (90% confidence interval) in various sex-age groups ranged from 0.023 (0.021 to 0.023) ng/kg of body weight per day for 30- to 45-year-old men to 0.382 (0.354 to 0.386) ng/kg of body weight per day for 2- to 4-year-old girls at the 99th percentile. The cancer risk of AFM1 to the general population of China was assessed to be 0.129 cancer cases per year per 10(8) persons at the 99th percentile. These results indicate that the health risk associated with AFM1 in milk in China is relatively low.

Topics: Adolescent; Adult; Aflatoxin M1; Age Factors; Aged; Animals; Body Weight; Carcinogens, Environmental; Cattle; Child; Child, Preschool; China; Dairy Products; Female; Food Contamination; Humans; Male; Middle Aged; Milk; Monte Carlo Method; Risk Assessment; Yogurt; Young Adult

Haemato- and myelotoxicity are adverse effects caused by mycotoxins. Due to the relevance of aflatoxins to human health, the present study, employing CFU-GM-, BFU-E- and CFU-E-clonogenic assays, aimed at (i) comparing, in vitro, the sensitivity of human vs. murine haematopoietic progenitors to AFB1 and AFM1 (0.001-50microg/ml), (ii) assessing whether a single AFB1 in vivo treatment (0.3-3mg/kgb.w.) alters the ability of murine bone marrow cells to form myeloid and erythroid colonies, and (iii) comparing the in vitro with the in vitro ex-vivo data. We demonstrated (i) species-related sensitivity to AFB1, showing higher susceptibility of human myeloid and erythroid progenitors (IC(50) values: about 4 times lower in human than in murine cells), (ii) higher sensitivity of CFU-GM and BFU-E colonies, both more markedly affected, particularly by AFB1 (IC(50): 2.45+/-1.08 and 1.82+/-0.8microM for humans, and 11.08+/-2.92 and 1.81+/-0.20microM for mice, respectively), than the mature CFU-E (AFB1 IC(50): 12.58+/-5.4 and 40.27+/-6.05microM), irrespectively of animal species, (iii) regarding AFM1, a species- and lineage-related susceptibility similar to that observed for AFB1 and (iv) lack of effects after AFB1 in vivo treatment on the proliferation of haematopoietic colonies.

Topics: Aflatoxin B1; Aflatoxin M1; Animals; Blood Cell Count; Body Weight; Bone Marrow Cells; Cell Lineage; Cell Separation; Colony-Forming Units Assay; Erythroid Precursor Cells; Hematopoietic Stem Cells; Humans; Leukocytes; Male; Mice; Mutagens; Species Specificity

The aim was to carry out a survey of aflatoxin M(1) (AFM(1)) in raw whole milk from bulk tanks. The sample collection was performed in farms located in one the most important milk-production zones in the centre of Argentina. A total of 94 samples of milk from 47 dairy farms were analysed. AFM(1) analysis involved the use of liquid chromatography-tandem mass spectrometry (LC-MS/MS) with prior purification of the extracts using immunoaffinity columns. AFM(1) incidence in raw milk was high as 63.8% and levels were between not detected to 0.07 microg l(-1). Several contaminated samples (39%) were over the European Commission limit for infant milk (0.025 microg l(-1)), although none of samples were above Argentine legislation. Estimates of AFM(1) intake were assessed for different age populations. The average AFM(1) estimated daily intakes were 1.6, 0.5, 0.17 ng kg(-1) body weight day(-1) for 4-year-old babies, young children, and adults, respectively. All tested farms used pastures and silages at similar composition. Even though some farms (13) employed high-risk supplementary feeds, such as peanut pod and/or cotton seed, no statistically significant differences were observed between groups. Information from AFM(1) levels in milk in Argentina is limited. A systematic AFM(1) monitoring programme must be performed by means of accurate and reliable analytical techniques as a strategy for protecting milk consumers.

Topics: Adult; Aflatoxin M1; Agriculture; Animal Feed; Animals; Argentina; Body Weight; Cattle; Child, Preschool; Food Contamination; Humans; Infant; Milk

In one experiment, the effect of inorganic sorbents on the metabolic fate of aflatoxin B1 (AFB1) was studied in turkey poults. At 5 weeks of age, female poults were surgically colostomized and 9 days later orally dosed with 0.75 mg AFB1/kg BW. Hydrated sodium calcium aluminosilicate (HSCAS), acidic HSCAS, and activated charcoal (AC) were tested, by concomitant administration with AFB1. Urine was collected up to 48 h post-dosing and analyzed for aflatoxin M1 (AFM1) which was the major metabolite found in all treatment groups. Hydrated sodium calcium aluminosilicate, previously proven beneficial in alleviating aflatoxicosis in farm animals, reduced urinary AFM1 output when orally dosed simultaneous with AFB1. Also, acidic HSCAS and AC significantly decreased AFM1 excretion when administered concomitantly with AFB1. A second experiment was conducted to evaluate the ability of two types of AC to modify aflatoxicosis when added to aflatoxin (AF)-contaminated (from culture material) diets of turkey poults. Although AC was able to decrease AFM1 excretion in the first experiment, no protective effects from AF toxicity were observed in the feeding study.

Topics: Aflatoxin M1; Aluminum Silicates; Animal Feed; Animals; Blood Chemical Analysis; Body Weight; Charcoal; Female; Mycotoxicosis; Organ Size; Turkeys

When aflatoxin-contaminated grain is consumed by dairy cows, aflatoxin M1 is excreted in the milk. Sixteen neonatal male Holstein calves were given milk which had been collected from cows given 5 to 6 mg of aflatoxin B1 each day. The calves were examined for possible detrimental effects of the mycotoxin at pseudophysiologic concentrations. Calves were allotted to 1 of 4 groups given different milk dietary aflatoxin M1 concentrations: group 1--given 0 microgram of aflatoxin M1/L (undetectable); group 2--given 0.5 microgram/L; group 3--given 1 microgram/L; and group 4--given 2 micrograms/L. Whole milk equal to 8% of body weight was fed daily and adjusted each week to maintain this ratio. Water and a 15% crude protein complete calf starter ration were offered ad libitum for the 6-week feeding study. Weekly blood samples were collected via jugular venipuncture and analyzed for serum alkaline phosphatase and aspartate aminotransferase activities. Daily means for milk dry matter intake (in kg) and complete ration intake (in kg) for the calf groups were as follows: 0.46 and 0.36 for group 1; 0.46 and 0.25 for group 2; 0.42 and 0.18 for group 3; and 0.49 and 0.40 for group 4. Significant differences in complete ration and total dry matter intake were noted. The average daily gains (in kg) and gains in height at withers (in cm) were 0.39 and 4.1 for group 1; 0.36 and 4.0 for group 2; 0.29 and 5.7 for group 3; and 0.42 and 5.1 for group 4.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Aflatoxin M1; Aflatoxins; Alkaline Phosphatase; Animals; Animals, Newborn; Aspartate Aminotransferases; Body Weight; Cattle; Eating; Female; Food Contamination; Lactation; Male; Milk; Organ Size; Pregnancy

Eight male Holstein calves were raised to 6 wk of age on whole milk at 8% of body weight and calf starter ration (15% crude protein) free choice. For each of 2 days prior to sacrifice, four of the calves were selected at random and administered the mixed function oxidase inducer beta-napthoflavone by bolus in starch carrier at .005% of metabolic body weight. After sacrifice, 10,000 X g postmitochondrial supernatant preparations were made from the excised livers and incubated with a 50-micrograms challenge of aflatoxin B1. Although there was no significant difference in total percent of the challenge metabolized, there was a large increase in aflatoxin M1 production by liver preparations from calves administered the flavone inducer. If the flavone family of inducers could alter the metabolic profile for aflatoxin in the adult lactating bovine, aflatoxin M1 excretion rates could be increased and pathways for production of mutagenic metabolites could be enhanced.

Topics: Aflatoxin B1; Aflatoxin M1; Aflatoxins; Animals; Benzoflavones; beta-Naphthoflavone; Body Weight; Carcinogens; Cattle; Cell-Free System; Flavonoids; Fluorometry; Liver; Male; Organ Size