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PubMed2006

Influence of mycotoxin zearalenone and its derivatives (alpha and beta zearalenol) on apoptosis and proliferation of cultured granulosa cells from equine ovaries.

BACKGROUND: The mycotoxin zearalenone (ZEA) and its derivatives, alpha and beta-zearalenol (alpha and beta-ZOL), synthesized by genera Fusarium, often occur as contaminants in cereal grains and animal feeds. The importance of ZEA on reproductive disorders is well known in domestic animals species, particularly in swine and cattle. In the horse, limited data are available to date on the influence of dietary exposure to ZEA on reproductive health and on its in vitro effects on reproductive cells. The aim of this study was to evaluate the effects of ZEA and its derivatives, alpha and beta-ZOL, on granulosa cells (GCs) from the ovaries of cycling mares. METHODS: The cell proliferation was evaluated by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test after 3 days exposure at different concentrations of ZEA and its derivatives (from 1 x 10-7 to 0.1 microM). The apoptosis induction was evaluated after 1 day exposure, by DNA analysis using flow cytometry. RESULTS: An increase in cell proliferation with respect to the control was observed in the presence of ZEA at 1 x 10-3 and 1 x 10-4 microM and apoptosis was induced by all mycotoxins at different concentrations. CONCLUSION: The simultaneous presence of apoptosis and proliferation in GC cultures treated with zearalenones could indicate that these mycotoxins could be effective in inducing follicular atresia. These effects of zearalenones may result from both direct interaction with oestrogen-receptors as well as interaction with the enzymes 3alpha (beta)-hydroxysteroid dehydrogenase (HSD), involved in the synthesis and metabolism of endogenous steroid hormones. These cellular disturbances, described for the first time in equine GCs cultured in vitro, could be hypothesized as referred to reproductive failures of unknown ethiology in the mare.

Animal Feed↗

PubMed1995

Trichothecene mycotoxins depress the mononuclear-phagocytic system of young turkeys.

Macrophage cells isolated from the abdominal cavity of 21-day-old turkeys after a single injection of Sephadex suspension were used to quantitate the effects of direct in vitro exposure to deoxynivalenol (DON), 3-acetyldeoxynivalenol (3ac-DON), scirpentriol (STO), or 15-acetylscirpenol (15-MAS). Macrophage monolayers were established on glass surfaces and cells were exposed to graded levels of individual mycotoxins for 1 hour: DON, 20-640 micrograms/microliters of culture; 3ac-DON, STO, 15-MAS, 20-1280 micrograms/microliters of culture. All four mycotoxins caused dose-related effects. A concentration of 50 micrograms/microliter DON caused a significant decrease in macrophage adherence, phagocytosis of opsonized SRBC, and number of opsonized SRBC per macrophage; at 200 micrograms/microliter, phagocytosis of unopsonized SRBC was decreased. There were also increasing percentages of damaged macrophages with increasing DON doses as indicated by morphological alterations. Linear decreases in macrophage viability on exposure to 3-acDON and STO were observed. Moreover, STO and 15-MAS decreased macrophage adherence to glass and 3-acDON, STO, and 15-MAS induced macrophage morphological alterations. This study suggests that trichothecene mycotoxins may be immunosuppressive by affecting viability, adherence and phagocytic potential of mononuclear phagocytic cells of young turkeys.

Animals↗

PubMed2004

Detection of trichothecene mycotoxins in sera from individuals exposed to Stachybotrys chartarum in indoor environments.

To date, no study has effectively demonstrated a direct human exposure to mycotoxins in mold-contaminated buildings. Therefore, the authors investigated the presence of trichothecene mycotoxins in sera from individuals exposed to indoor molds (specifically Stachybotrys chartarum). Sera from occupants of contaminated (test samples, n=44) and uncontaminated (control samples, n=26) buildings were analyzed using a competitive enzyme-linked immunosorbent assay (ELISA) highly specific for macrocyclic trichothecenes. Twenty-three samples were significantly different (p < 0.05) from normal human serum tested in the same manner, whereas only 1 of the control samples tested positive. Mass spectrometry analysis could not confirm the presence of intact S. chartarum macrocyclic trichothecenes. The authors hypothesize that this result was caused by uncharacterized ELISA-reactive metabolic breakdown products. Data from this study suggest that trichothecene mycotoxins can be demonstrated in the tissues of certain individuals exposed to S. chartarum in contaminated buildings.

Air Pollution, Indoor↗

PubMed2003

[Experimental lung carcinoma induced by fungi and mycotoxins--a review].

Up to now, most of the etiological studies of lung cancer have been focused on air pollution, radiation and smoking, etc, whereas in the past decades, we had found that fungi and mycotoxins in foodstuff might play important roles in the carcinogenesis of lung cancer. Our preliminary work showed that the liquid in pickled vegetables consumed by residents of Taihang Mountain area in Hebei Province and corn flours inoculated with Aspergillus flavus, Aspergillus versicolor in which aflatoxin G1(AFG1) and Sterigmatocystin (ST) were detected, could induce lung adenocarcinoma in Kunming, LACA and NIH mice as well as Wistar rats. Further studies with AFG1 and ST confirmed that oral administration of the two mycotoxins could induce lung adenocarconma in NIH mice. The experimental lung carcinoma induction studies with fungi and mycotoxins in our lab are summarized in this review.

Aflatoxins↗

PubMed2001

[Natural contamination with mycotoxins in forage maize and green coffee in Nayarit State (Mexico)].

The presence of mycotoxins in forage maize (zearalenone, fumonisin B1, T-2 toxin and diacetoxyscirpenol) and green coffee (ochratoxin A) from Nayarit State (Mexico) has been studied. All maize samples analyzed showed fumonisin B1 contamination, with an average concentration of 2,541 microg/kg. Fifteen percent of the samples contained zearalenone, with an average concentration of 1,610 microg/kg. Only one sample showed T-2 toxin contamination (7 microg/kg), and no diacetoxyscirpenol was detected. Sixty-seven per cent of green coffee samples were contaminated with ochratoxin A, with an average concentration of 30.1 microg/kg. This is the first study about mycotoxins developed in Nayarit and it has shown that mycotoxin contamination is a real problem in both foodstuffs studied.

English Abstract↗

PubMed2000

[Emerging mycotoxins: introduction].

Mycotoxins are a diverse group of secondary metabolites produced by moulds. They have a wide range of toxicological effects both in humans and animals. Nowadays about 200 of these metabolites have been described, but only a few of them may be considered as important from the agricultural point of view. In this publication, some aspects such as mycotoxin producing species, analytical methods, natural occurrence, preventive and detoxification techniques and regulations of these kind of mycotoxins will be developed by different authors involved in these research issues.

English Abstract↗

PubMed1983

[Effects of various antibiotics and natural mycotoxins on the hematopoietic stem cells of the bone marrow in normal and adjuvant-treated rats].

This experiment was carried out, in order to investigate the effect of antibiotics and natural mycotoxin on the hematopoietic stem cells at the normal and inflammatory condition. Adjuvant-treated rats (Aj-rats) are considered as a model of human rheumatoid arthritis. We measured the CFU-C and CFU-E of bone marrow of normal and Aj-rats which were injected with large (1.0 g/kg X 3) and small doses (0.5 g/kg X 3) of ampicillin (ABPC), cefazolin (CEZ), chloramphenicol (CP) and fusarenon-X (F-X). In Aj-rats the number of CFU-C was 1.5 times higher and CFU-E 60% less than normal. Injection of large doses of ABPC enhanced markedly the numbers of CFU-C in Aj-rats and suppressed slightly CFU-E in normal rats. Large doses of CEZ inclined to increase CFU-C and decreased CFU-E in normal and Aj-rats. Injection of small doses of CP tended to increase CFU-C and to decrease CFU-E, and large doses of CP to suppress both CFU-C and CFU-E levels in normal or Aj-rats. F-X, natural mycotoxin suppressed markedly both CFU-C and CFU-E levels of normal rats, and slightly the CFU-E in Aj-rats. These results suggest that one should pay attention to the fact that some doses of antibiotics or natural mycotoxin might be harmful on the bone marrow hematopoietic stem cells.

Ampicillin↗

PubMed1996

Mitogenic effects of mycotoxins on T4 lymphocytes.

Mycotoxins have compromising effects on varied biological systems, even when ingested at levels which do not evoke manifestations of clinical mycotoxicoses. No data have been previously found as to the therapeutic and mitogenic effects of mycotoxins. This was the objective of the present work. Human peripheral T4 lymphocytes were obtained by venipuncture, propagated in RPMI 1640 medium and challenged with varied concentrations of aflatoxins, B1, B2, G1, and G2. The cells were stained with propidium iodide and fluorescent isothiocyanate (FITC) and examined microfluorometrically. All the mycotoxins were significantly mitogenic on the basis of dose response. No adverse effects were observed when doses of 10, 50 and 100 micrograms were administered on a modest clinical basis to volunteers.

Aflatoxins↗

PubMed1999

Overview of mycotoxin methods, present status and future needs.

This article reviews current requirements for the analysis for mycotoxins in foods and identifies legislative as well as other factors that are driving development and validation of new methods. New regulatory limits for mycotoxins and analytical quality assurance requirements for laboratories to only use validated methods are seen as major factors driving developments. Three major classes of methods are identified which serve different purposes and can be categorized as screening, official and research. In each case the present status and future needs are assessed. In addition to an overview of trends in analytical methods, some other areas of analytical quality assurance such as participation in proficiency testing and reference materials are identified.

Chemistry, Analytic↗

PubMed1999

Fiber-optic immunosensor for mycotoxins.

Evanescent wave-based fiber-optic immunosensors were studied for the detection of fumonisins and aflatoxins in maize. Two formats, competitive and non-competitive, were used. A competitive format was used to measure fumonisin B1 (FB1) in both spiked and naturally contaminated maize samples. Fumonisin monoclonal antibodies were covalently coupled to an optical fiber and the competition between FB1 and FB1 labeled with fluorescein (FB1-FITC) for the limited number of binding sites on the fiber was assessed. The signal generated in the assay was inversely proportional to the FB1 concentration. For samples, the concentration causing an inhibition of binding by 50% (IC50) was dependent upon the clean-up procedure used. Simple dilution of methanolic maize extracts yielded an assay with an IC50 equivalent to 25 microg FB1 g(-1) maize with a limit of detection of 3.2 microg g(-1) maize. Affinity column clean-up yielded an assay with an IC50 equivalent to 5 microg FB1 g(-1) maize (limit of detection 0.4 microg FB1 g(-1)). An HPLC method and the immunosensor method agreed well for naturally contaminated maize samples except when large amounts of other fumonisins that cross-react with the immunosensor were present. The second sensor format, for the mycotoxin aflatoxin B1 (AFB1), was a non-competitive assay using the native fluorescence of this mycotoxin. Because the fluorescence of AFB1 itself was detected, the response of the sensor was directly proportional to the toxin concentration. The sensor, while capable of detecting as little as 2 ng ml(-1) of AFB1 in solution was technically not an immunosensor, since the attachment of aflatoxin specific antibodies was not required. Sensors of the formats described have the potential to rapidly screen individual maize samples but require coupling with a clean-up technique to be truly effective.

Aflatoxin B1↗

PubMed1987

Comparison of thermally-assisted fast atom bombardment (TA-FAB) with conventional FAB and EI mass spectrometry for the analysis of the Helminthosporium carbonum mycotoxins.

A new technique known as thermally-assisted fast atom bombardment (TA-FAB) has been applied to the analysis of a series of cyclic tetrapeptide mycotoxins in order to demonstrate the usefulness of the method for structural elucidation. TA-FAB uses saturated aqueous solutions of highly hydroxylated compounds, such as fructose, as alternatives to the usual viscous liquid matrices employed in conventional FAB. During the TA-FAB analysis, the probe tip is resistively heated causing differences to occur in the desorption profiles for the analyte and the matrix ions enabling an optimization for analyte desorption as a function of temperature. In this study, direct comparisons are made between TA-FAB, conventional FAB, and electron impact ionization for the analysis of the Helminthosporium carbonum mycotoxins at the 1.5 micrograms level. The results demonstrate the superior capacity of TA-FAB to provide both molecular weight confirmation and significant fragmentation to aid in the structural elucidation of these important biomolecules.

Helminthosporium↗

PubMed2003

Occurrence of certain mycotoxins in corn and corn-based products and thermostability of fumonisin B1 during processing.

A total of 57 samples of corn and corn-based products collected from various districts of Egypt were analyzed for Fusarium mycotoxins (T-2, diacetoxyscripenol (DAS), deoxynivalenol (DON) and fumonisin B1 (FB1) and aflatoxins. FB1 was detected in about 80%, 53.85%, 33.3%, and 28.57% of yellow corn, corn meal, white corn, and popcorn samples, respectively. The levels of FB1 ranged from 10 to 780 microg/kg. T-2 and DAS were detected in 5% and 10% of yellow corn samples, respectively, and DON was detected in white corn and popcorn samples at levels of 28.8 and 10.1 microg/kg, respectively. Starch samples were found to be free from Fusarium mycotoxins. Baking balady bread at 450 degrees C/min reduced FB1 to 72.4% while baking Franco bread at 250 degrees C/20 min reduced FB1 to 57.4%. Boiling of macaroni and corn in water completely removed FB1 from contaminated samples. On the other side, corn flakes samples were found to be contaminated with aflatoxins B1 and G1 at levels ranging from 6 to 10 ppm, whereas 2.9% of samples were contaminated with aflatoxin B1 > 35 ppm and G1 > 16 ppm.

Egypt↗

PubMed1984

Studies on the mechanism of toxicity of the mycotoxin sporidesmin. 3--Inhibition by metals of the generation of superoxide radical by sporidesmin.

The mycotoxin sporidesmin has previously been shown to generate superoxide radical. This reaction involves autoxidation of the reduced form of the mycotoxin, a dithiol. In the present study, a number of mercaptide-forming metals have been shown to inhibit superoxide formation from sporidesmin in vitro. Furthermore, these metals decreased the rate of sporidesmin-induced hydrogen peroxide formation in erythrocytes and ameliorated the subsequent oxidative damage to these cells. These effects were found to be specific to sporidesmin; mercaptide-forming metals did not inhibit the changes induced by compounds which are not dependent upon thiol groups for active oxygen generation. Zinc was one of the most potent inhibitors of superoxide generation from sporidesmin in these test systems; only mercury and cadmium were significantly more active. Salts of zinc are known to provide effective protection against the harmful effects of sporidesmin in vivo. The results of these studies provide a possible explanation for this effect.

Animals↗

PubMed1993

Toxigenic fungi and mycotoxin content in poultry feedstuff ingredients.

Fifty isolates of Aspergillus flavus as well as 25 isolates of each of A. fumigatus, A. ochraceus, A. terreus and Fusarium spp. isolated from poultry feeds were screened for toxin production. Of these isolates, 20% have been detected to exhibit toxigenic activity. The mycotoxin content was determined in samples of grind maize, cotton-seed cake, fish meal, wheat bran and soybean meal. 80% of maize samples were highly contaminated with aflatoxins (480 micrograms/kg) and zearalenone (40 micrograms/kg). Aflatoxins B1 and B2 were found in 50 and 30% of cotton-seed cake and fish meal, respectively. With wheat bran, 40% of the samples contained aflatoxin B1 and zearalenone. Soybean meal appeared to be a poor substrate for mycotoxin formation.

Aflatoxins↗

PubMed1993

Survey of mycoflora and mycotoxins of some dried fruits in Egypt.

Fifty-five species and two varieties appertaing to 23 genera were collected and identified from 4 samples of dried fig and 3 dried samples of each of apricot, plum and raisin. Forty-nine species and two varieties belonging to 20 genera were isolated on 1% glucose-Czapek's while 31 species and one variety belonging to 16 genera were isolated on 40% sucrose-Czapek's agar medium at 28 +/- 2 degrees C. Penicillium, Aspergillus and cladosporium were the common genera on the two types of media used. Alternaria and Pleospora were common on 1% glucose-Czapek's agar only while the genus of Eurotium was isolated with high occurrence on 40% sucrose-Czapek's agar. The common species on the two types of media used were Penicillium chrysogenum, Aspergillus niger and cladosporium cladosporioides. On 1% glucose-Czapek's agar, P. aurantiogriseum, A. fumigatus, A. flavus, Alternaria tenuissima and Pleospora herbarum were isolated with high occurrence while A. versicolor, A. wentii, Eurotium amstelodami and E. chevalieri were common on 40% sucrose-Czapek's agar. The different dried fruit samples were assayed for the natural occurrence of aflatoxins B1, B2, G1 & G2, citrinin, ochratoxins, patulin, sterigmatocystin, diacetoxyscirpenol, T-2 toxin and zearalenone by thin layer chromatographic analysis. Ochratoxin A was detected in all samples tested of apricot (50-110 micrograms/kg), fig (60-120 micrograms/kg) and plum (210-280 micrograms/kg). The other mycotoxins under investigation were not detected. All samples examined of raisin proved to be mycotoxin free.

Egypt↗

PubMed2006

Fusarium mycotoxins and ochratoxin A in cereals and cereal products: results from the Bavarian Health and Food Safety Authority in 2004.

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.

Edible Grain↗

PubMed2026

Sex-Specific Renal Proteomic Responses to Mycotoxins and Their Mitigation by Bioactive Food Ingredients.

This study aimed to evaluate sex-related differences in kidney toxicity induced by aflatoxin B1 (AFB1) and ochratoxin A (OTA), administered either individually or in combination, and to investigate the mitigating potential of fermented whey (FW) and pumpkin (P) using a proteomic approach in a sub-chronic rat model. Diets naturally contaminated with AFB1 and OTA fungal producers, or enriched with FW and P, were administered to 120 Wistar rats for 28 days. Afterwards, kidneys were collected and subjected to protein extraction and digestion to peptides. For proteomic analysis, peptides were separated using an LC-QTOF-MS system, and differentially expressed proteins between control and treated groups were statistically filtered (p < 0.05) to distinguish sex-related differences between males and females. Proteomic analysis revealed that non-supplemented diets induced significant protein alterations, with male kidneys showing greater sensitivity to mycotoxin exposure, particularly to OTA, and exhibiting a distinct expression pattern compared with females. Notably, FW and FW + P supplementation mitigated these adverse effects by reducing the number of differentially expressed proteins, stabilizing mitochondrial function, and enhancing xenobiotic metabolism pathways. These findings underscore the importance of sex-specific factors in evaluating the protective potential of bioactive ingredients against mycotoxin-induced renal injury.

Animals↗

PubMed1993

Rapid colorimetric bioassay for screening of Fusarium mycotoxins.

The cytotoxicity of Fusarium mycotoxins was evaluated using a trichothecene sensitive cell line (BHK-21, baby hamster kidney cells) in combination with the MTT-cleavage test as an end-point measurement. Cells tended to be more sensitive to the type A trichothecenes with midpoint cytotoxicity values ranging from 1.6 ng/ml for T-2 toxin to 60 ng/ml for scirpentriol. The cytotoxicity value for deoxynivalenol (type B) was 112 ng/ml. The inherent disadvantage of the MTT-assay (formation of insoluble formazan) was overcome by using the analog MTS and measuring the water-soluble formazan directly in the culture media. The MTS-midpoint cytotoxicity values for T-2 toxin and deoxynivalenol (2.1 and 141 ng/ml, respectively), although slightly higher, showed a good correspondence to the MTT-test. Both the MTT- and MTS-cleavage tests are useful for evaluating the cytotoxicity of Fusarium mycotoxins. The replacement of MTT by MTS substantially reduced the number of sample processing steps and the length of time required to complete the cytotoxicity assay.

Animals↗