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Toxins May 2015The intestinal epithelium is the first barrier against food contaminants. Zearalenone (ZEN) is an estrogenic mycotoxin that was identified as a common contaminant of...
The intestinal epithelium is the first barrier against food contaminants. Zearalenone (ZEN) is an estrogenic mycotoxin that was identified as a common contaminant of cereal grains and food and feedstuffs. In the present study, we have investigated the in vitro effects of ZEN and some of its metabolites (α-ZOL, β-ZOL) in concentrations of 10-100 µM on a swine epithelial cell line: Intestinal porcine epithelial cells (IPEC-1). We demonstrated that both ZEN metabolites were more toxic for IPEC cells as resulted from the XTT test, while for doses lower than 10 µM, only β-ZOL showed a more pronounced cytotoxicity versus epithelial cells as resulted from neutral red assay. ZEN has no effect on TER values, while α-ZOL significantly decreased the TER values, starting with day 4 of treatment. β-ZOL had a dual effect, firstly it induced a significant increase of TER, and then, starting on day 6, it induced a dramatic decrease of TER values as compared with on day 0. Concerning the cytokine synthesis, our results showed that ZEN has a tendency to increase the synthesis of IL-8 and IL-10. By contrast, α- and β-ZOL decreased the expression of both IL-8 and IL-10, in a dose dependent manner. In conclusion, our results showed that ZEN and its metabolites differently affected porcine intestinal cell viability, transepithelial resistance and cytokine synthesis with important implication for gut health.
Topics: Animals; Cell Line; Cell Survival; Epithelial Cells; Estrogens, Non-Steroidal; Food Contamination; Interleukin-10; Interleukin-8; Intestinal Mucosa; Swine; Zearalenone; Zeranol
PubMed: 26035492
DOI: 10.3390/toxins7061979 -
Toxins Jun 2018The potential effect of difructose anhydride III (DFA III) supplementation in cattle feed was evaluated using a previously developed urinary-zearalenone (ZEN) monitoring...
The potential effect of difructose anhydride III (DFA III) supplementation in cattle feed was evaluated using a previously developed urinary-zearalenone (ZEN) monitoring system. Japanese Black cattle from two beef herds aged 9⁻10 months were used. DFA III was supplemented for two weeks. ZEN concentrations in feed were similar in both herds (0.27 and 0.22 mg/kg in roughage and concentrates, respectively), and below the maximum allowance in Japan. ZEN, α-zearalenol (α-ZOL), and β-ZOL concentrations in urine were measured using LC/MS/MS the day before DFA III administration, 9 and 14 days thereafter, and 9 days after supplementation ceased. Significant differences in ZEN, α-ZOL, β-ZOL, and total ZEN were recorded on different sampling dates. The concentration of inorganic phosphate in DFA III-supplemented animals was significantly higher than in controls on day 23 (8.4 vs. 7.7 mg/dL), suggesting a possible role of DFA III in tight junction of intestinal epithelial cells. This is the first evidence that DFA III reduces mycotoxin levels reaching the systemic circulation and excreted in urine. This preventive effect may involve an improved tight-junction-dependent intestinal barrier function. Additionally, our practical approach confirmed that monitoring of urinary mycotoxin is useful for evaluating the effects of dietary supplements to prevent mycotoxin adsorption.
Topics: Animal Feed; Animals; Calcium; Cattle; Dietary Exposure; Dietary Supplements; Disaccharides; Environmental Monitoring; Female; Intestinal Absorption; Magnesium; Phosphates; Zearalenone
PubMed: 29857569
DOI: 10.3390/toxins10060223 -
Toxins Aug 2015Zearalenone (ZEA) and its derivatives are mycotoxins with estrogenic effects on mammals. The biotransformation for ZEA in animals involves the formation of two major...
Zearalenone (ZEA) and its derivatives are mycotoxins with estrogenic effects on mammals. The biotransformation for ZEA in animals involves the formation of two major metabolites, α- and β-zearalenol (α-ZOL and β-ZOL), which are subsequently conjugated with glucuronic acid. The capability of Saccharomyces cerevisiae strains isolated from silage to eliminate ZEA and its derivatives α-ZOL and β-ZOL was investigated as, also, the mechanisms involved. Strains were grown on Yeast Extract-Peptone-Dextrose medium supplemented with the mycotoxins and their elimination from medium was quantified over time by HPLC-FL. A significant effect on the concentration of ZEA was observed, as all the tested strains were able to eliminate more than 90% of the mycotoxin from the culture medium in two days. The observed elimination was mainly due to ZEA biotransformation into β-ZOL (53%) and α-ZOL (8%) rather than to its adsorption to yeast cells walls. Further, the biotransformation of α-ZOL was not observed but a small amount of β-ZOL (6%) disappeared from culture medium. ZEA biotransformation by yeasts may not be regarded as a full detoxification process because both main end-products are still estrogenic. Nonetheless, it was observed that the biotransformation favors the formation of β-ZOL which is less estrogenic than ZEA and α-ZOL. This metabolic effect is only possible if active strains are used as feed additives and may play a role in the detoxification performance of products with viable S. cerevisiae cells.
Topics: Adsorption; Animal Feed; Animals; Biotransformation; Cattle; Saccharomyces cerevisiae; Zearalenone
PubMed: 26308051
DOI: 10.3390/toxins7083297 -
Toxins Mar 2019Mycotoxins are fungal secondary metabolites that pose health risks to exposed individuals, requiring necessary measures to reduce them. Using liquid...
Mycotoxins are fungal secondary metabolites that pose health risks to exposed individuals, requiring necessary measures to reduce them. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), mycotoxins were quantified in whole grain sorghum and subsequently derived from two sorghum varieties (high and low tannin). The whole grain (WG) samples were obtained by fermenting sorghum with strains (FUA 3165 and FUA 3321). Naturally (spontaneously) fermented WG- under the same conditions were equally analysed. Among the mycotoxins investigated, fumonisin B₁ (FB₁), B₂ (FB₂), B₃ (FB₃), T-2 toxin (T-2), zearalenone (ZEA), alpha-zearalenol (α-ZOL) and beta-zearalenol (β-ZOL) were detected in sorghum. Results obtained showed that mycotoxin concentrations significantly ( ≤ 0.05) reduced after fermentation. In particular, FUA 3321 showed the capability to significantly ( ≤ 0.05) reduce all the mycotoxins by 98% for FB₁, 84% for T-2 and up to 82% for α-ZOL, compared to raw low tannin sorghum. Fermenting with the strains showed potential to effectively reduce mycotoxin contamination in whole grain . Thus, we recommended FUA 3321 in particular to be used as a potential starter culture in sorghum fermentation.
Topics: Fermentation; Food Contamination; Limosilactobacillus fermentum; Mycotoxins; Sorghum; South Africa; Whole Grains
PubMed: 30934589
DOI: 10.3390/toxins11030180 -
Food Chemistry Jan 2023The aim of this manuscript was to validate and apply an analytical methodology for the simultaneous determination of 34 mycotoxins in cocoa. The extraction method used...
The aim of this manuscript was to validate and apply an analytical methodology for the simultaneous determination of 34 mycotoxins in cocoa. The extraction method used in the tests was a liquid-liquid partition by NaCl addition with a freezing step followed by quantification using LC-MS/MS. The results were discussed based on national and international directives for food contaminants. The recoveries and precision were adequate, except for the mycotoxins ionized with the ammonium adduct (NH), E-cristinine and β-ZOL. This result directly influenced the measurement uncertainty of these mycotoxins, because the precision and the correction factor of the recovery were the factors with the greatest impact on the uncertainty of the method. The evaluation of the matrix effect showed considerable signal suppression for 53 % of the evaluated mycotoxins. Nevertheless, the mycotoxins exhibited relatively low quantification limits, with values between 1 and 75 μg kg. The validated methodology was applied to 15 cocoa samples collected in warehouses in Brazil. Positive results were found for all the evaluated samples, in which nine toxins were detected out of the 34 investigated.
Topics: Cacao; Chromatography, High Pressure Liquid; Chromatography, Liquid; Mycotoxins; Tandem Mass Spectrometry; Uncertainty
PubMed: 36027808
DOI: 10.1016/j.foodchem.2022.133902 -
Toxins Jan 2014Human exposure assessment to deoxynivalenol (DON), aflatoxin B1 (AFB1), fumonisin B1 (FB1), zearalenone (ZEA) and ochratoxin A (OTA) can be performed by measuring their...
Human exposure assessment to deoxynivalenol (DON), aflatoxin B1 (AFB1), fumonisin B1 (FB1), zearalenone (ZEA) and ochratoxin A (OTA) can be performed by measuring their urinary biomarkers. Suitable biomarkers of exposure for these mycotoxins are DON + de-epoxydeoxynivalenol (DOM-1), aflatoxin M1 (AFM1), FB1, ZEA + α-zearalenol (α-ZOL) + β-zearalenol (β-ZOL) and OTA, respectively. An UPLC-MS/MS multi-biomarker method was used to detect and measure incidence and levels of these biomarkers in urine samples of 52 volunteers resident in Apulia region in Southern Italy. The presence of ZEA + ZOLs, OTA, DON, FB1 and AFM1 were detected in 100%, 100%, 96%, 56% and 6%, of samples, respectively. All samples contained biomarkers of two or more mycotoxins. The mean concentrations of biomarkers ranged from 0.055 ng/mL (FB1) to 11.89 ng/mL (DON). Urinary biomarker concentrations were used to estimate human exposure to multiple mycotoxin. For OTA and DON, 94% and 40% of volunteers, respectively exceeded the tolerable daily intake (TDI) for these mycotoxins. The estimated human exposure to FB1 and ZEA was largely below the TDI for these mycotoxins for all volunteers.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Biomarkers; Child; Child, Preschool; Chromatography, Liquid; Environmental Monitoring; Female; Humans; Italy; Male; Middle Aged; Mycotoxins; Tandem Mass Spectrometry; Young Adult
PubMed: 24476712
DOI: 10.3390/toxins6020523 -
International Journal of Molecular... Sep 2018Zearalenone hydrolase (ZHD) is the only reported α/β-hydrolase that can detoxify zearalenone (ZEN). ZHD has demonstrated its potential as a treatment for ZEN...
Zearalenone hydrolase (ZHD) is the only reported α/β-hydrolase that can detoxify zearalenone (ZEN). ZHD has demonstrated its potential as a treatment for ZEN contamination that will not result in damage to cereal crops. Recent researches have shown that the V153H mutant ZHD increased the specific activity against α-ZOL, but decreased its specific activity to β-ZOL. To understand whyV153H mutation showed catalytic specificity for α-ZOL, four molecular dynamics simulations combining with protein network analysis for wild type ZHD α-ZOL, ZHD β-ZOL, V153H α-ZOL, and V153H β-ZOL complexes were performed using Gromacs software. Our theoretical results indicated that the V153H mutant could cause a conformational switch at the cap domain (residues Gly161⁻Thr190) to affect the relative position catalytic residue (H242). Protein network analysis illustrated that the V153H mutation enhanced the communication with the whole protein and residues with high betweenness in the four complexes, which were primarily assembled in the cap domain and residues Met241 to Tyr245 regions. In addition, the existence of α-ZOL binding to V153H mutation enlarged the distance from the O atom in α-ZOL to the NE2 atom in His242, which prompted the side chain of H242 to the position with catalytic activity, thereby increasing the activity of V153H on the α-ZOL. Furthermore, α-ZOL could easily form a right attack angle and attack distance in the ZHD and α-ZOL complex to guarantee catalytic reaction. The alanine scanning results indicated that modifications of the residues in the cap domain produced significant changes in the binding affinity for α-ZOL and β-ZOL. Our results may provide useful theoretical evidence for the mechanism underlying the catalytic specificity of ZHD.
Topics: Amino Acid Substitution; Hydrolases; Hypocreales; Molecular Docking Simulation; Molecular Dynamics Simulation; Point Mutation; Protein Binding; Protein Conformation; Protein Stability; Zearalenone; Zeranol
PubMed: 30231501
DOI: 10.3390/ijms19092808 -
Toxicology in Vitro : An International... Apr 2018Epidemiological studies show that there is global decline in male fertility primarily as a result of poor sperm quality and this is attributed to exposure to endocrine...
Toxicological effects of regulated mycotoxins and persistent organochloride pesticides: In vitro cytotoxic assessment of single and defined mixtures on MA-10 murine Leydig cell line.
Epidemiological studies show that there is global decline in male fertility primarily as a result of poor sperm quality and this is attributed to exposure to endocrine disrupting chemicals (EDCs) in the environment, food and pharmaceutical products, including mycotoxins and pesticides. The Leydig cells in the male testes are responsible for producing androgens, hormones that play major roles in male development and reproductive function. Therefore, any toxin that affects the function and morphology of the Leydig cells may result in sub-fertility or infertility. The cytotoxic effects of single and binary mixtures of aflatoxin B (AFB), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEN), alpha-zearalenol (α-ZOL), beta-zearalenol (β-ZOL), 1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane (p,p'-DDT) and 1,1-dichloro-2,2-bis(p-chlorophenyl) ethylene (p,p'-DDE) on a model cell line, the MA-10 Leydig cells, were evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5-dipenyltetrazolium bromide (MTT) assay after 48h of exposure. With single toxin treatment at doses between 0.1μM and 64μM for 48h, DON was the most cytotoxic to MA-10 cells with a half maximal inhibitory concentration (IC) value of 12.3μM followed by α-ZOL (IC: 28μM) and OTA (IC: 30μM) while the IC of AFB, p,p'-DDT and p,p'-DDE were above the highest concentration tested (64μM). Co-exposure with p,p'-DDT or p,p'-DDE enhanced the toxicity of DON, OTA and ZEN to MA-10 Leydig cells, particularly at higher concentrations. This highlights the possible adverse effects on male reproductive health following co-exposure to these toxins.
Topics: Animals; Cell Line; Cell Survival; Endocrine Disruptors; Hydrocarbons, Chlorinated; Infertility, Male; Leydig Cells; Male; Mice; Mycotoxins; Pesticide Residues; Pesticides
PubMed: 29307701
DOI: 10.1016/j.tiv.2017.12.019 -
Bulletin of the World Health... 1972Gastric acid production, unstimulated and following stimulation with betazole hydrochloride, was measured in Indian men with cholera or acute vibrio-negative...
Gastric acid production, unstimulated and following stimulation with betazole hydrochloride, was measured in Indian men with cholera or acute vibrio-negative diarrhoea-Measurements were made during acute illness and after different periods of convalescence. Men from the same socioeconomic group and from a higher one served as controls. Stimulated acid production was severely reduced during diarrhoea caused by V. cholerae and related vibrios but not during acute vibrio-negative diarrhoea. Acid production returned to stable convalescent values 1-3 days after cessation of diarrhoea. Stimulated acid production was significantly lower in controls from the lower socioeconomic group than in those from the higher socioeconomic group. Achlorhydria that did not respond to betazole administration occurred in 32% of the convalescent cholera patients but in none of the controls or convalescent vibrio-negative diarrhoea patients. It is concluded from these results that diarrhoea produced by V. cholerae and related vibrios is accompanied by transient inhibition of gastric acid secretion, that cholera occurs largely in a population with impaired acid secretion, and that preexisting achlorhydria may predispose to infection with V. cholerae.
Topics: Acute Disease; Adolescent; Adult; Cholera; Convalescence; Diarrhea; Gastric Acidity Determination; Gastric Juice; Humans; India; Male; Middle Aged; Vibrio Infections
PubMed: 4538903
DOI: No ID Found -
Gastroenterology Jun 1988To assess the effect of malnutrition on gastric acidity and gastric bacterial colonization, we studied 35 severely malnourished Bangladeshi children before (0 wk) and...
To assess the effect of malnutrition on gastric acidity and gastric bacterial colonization, we studied 35 severely malnourished Bangladeshi children before (0 wk) and after (3 wk) they received nutritional rehabilitation for 3 wk. These results were compared with those obtained from a similarly examined group of 20 better-nourished Bangladeshi children. Gastric acid output, both basal and after betazole stimulation, was significantly lower in the malnourished group at 0 wk compared with the better-nourished children (p less than 0.01): basal 0.22 vs. 0.52 mEq HCl/h and stimulated 0.90 vs. 2.5 mEq HCl/h. Both the concentration of acid and the rate at which gastric juice was secreted were decreased in the malnourished group but serum gastrin levels were not significantly different. After 3 wk, the malnourished children had improved from 61% (+/- 9.0%; SD) to 81% (+/- 8.1%) of expected weight-for-height and were not significantly different than the better-nourished group (86% +/- 11%). Nevertheless, gastric acid concentration remained depressed in the 3-wk group, although the rate of gastric juice secretion equaled levels observed in the better-nourished group. None of the better-nourished children had detectable gram-negative bacterial colonization of their gastric juice. In contrast, 26 of 32 (81%) malnourished children at 0 wk were colonized--even after betazole stimulation, 11 of 33 (33%) gastric juice samples yielded viable organisms--suggesting that the decrease in gastric acid output greatly reduced the gastric acid barrier. Interestingly, only 9 of 20 (45%) better-nourished children had gastric juice with basal pH values below 4.0, suggesting that the gastric acid barrier may be an intermittent defense factor in Bangladeshi children.
Topics: Age Factors; Bacteria; Bangladesh; Body Weight; Child; Child, Preschool; Gastric Acid; Gastrins; Humans; Infant; Parenteral Nutrition, Total; Protein-Energy Malnutrition; Stomach
PubMed: 3129329
DOI: 10.1016/0016-5085(88)90668-3