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Subchronic feeding study of the mycotoxin fumonisin B1 in B6C3F1 mice and Fischer 344 rats.

Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium moniliforme, a common fungus which occurs naturally on corn, and other Fusarium species. FB1 and other fumonisins are now recognized as having potentially important animal and human health implications. However, few toxicological data are currently available. Male and female B6C3F1 mice and Fischer 344 rats were fed diets containing 0, 1, 3, 9, 27, or 81 ppm FB1 (> or = 98% purity) for 13 weeks. No differences in behavior or appearance, body weight or food consumption between control and FB1-fed groups were found. In mice, hepatopathy and altered serum chemical profiles indicative of hepatotoxicity were found in females fed the 81 ppm diet. No adverse effects were found in female mice fed < or = 27 ppm FB1 or in male mice at any dietary level studied. In rats, nephrosis involving the outer medulla was found in males fed > or = 9 ppm and, to a lesser degree, in females fed 81 ppm FB1, while decreased kidney weight was found in both sexes at dietary levels > or = 9 ppm FB1. Although the liver is a target organ of FB1 in rats, hepatotoxicity was not found in rats fed diets containing up to 81 ppm FB1 for 90 days. Thus, FB1 was toxic to both species following subchronic oral exposure, although significant interspecies differences in the no observed effect levels and organ-specific responses were found.

Adrenal Glands↗

Bisfuranoid mycotoxins: their genotoxicity and carcinogenicity.

Based on the mode of action of AFB1 and the activities of its biologically active intermediates, one may conclude that: 1. The mode of toxic action of the bisfuranoid mycotoxin is through epoxidation of the vinyl ether double bond of their dihydrobisfuran functionality. 2. The DNA and plasma albumin adducts formed in vivo may be useful in the molecular dosimetry of these environmental carcinogens. 3. There appears to be a linear correlation between the steady state levels of AFB1-FAPy-DNA adducts and the carcinogenicity of AFB1. Elucidation of the molecular basis of this correlation may shed light on the mechanism of AFB1-induced carcinogenesis. 4. Consistent appearance of AFB1-DNA adducts in the livers of liver cancer patients tested is supportive of the IARC conclusion that AFB1 is a human carcinogen involved in human primary liver cancer.

Aflatoxin B1↗

The mycotoxin fumonisin induces apoptosis in cultured human cells and in livers and kidneys of rats.

Fumonisin B1 is a mycotoxin produced by Fusarium moniliforme, a fungus that infects corn and other grains in the U.S. Fumonisin ingestion causes a variety of effects including equine leukoencephalomalacia and porcine pulmonary edema, and has been associated epidemiologically with human esophageal cancer. Fumonisin B1 produces growth inhibition and increased apoptosis in primary human keratinocyte cultures and in HET-1A cells. In order to set the doses for a 2-year tumor bioassay, male and female F344 rats were fed fumonisin B1 (99, 163, 234, and 484 ppm) for 28 days and the organs examined histologically. There was a dose dependent decrease in liver and kidney weights in the rats. The liver weight loss was accompanied by the induction of apoptosis and hepatocellular and bile duct hyperplasia in both sexes, with the female rats being more responsive at lower doses. The induction of tubular epithelial cell apoptosis was the primary response of the kidneys to dietary fumonisin B1. Apoptosis was present at all doses in the kidneys of the male rats, and occurred in the females only at 163, 234, and 484 ppm fumonisin B1. These results demonstrate that fumonisin B1 treatment causes a similar increase in apoptosis both in vivo and in vitro.

Animals↗

Mycotoxin induction of somatic mosaicism in Drosophila and DNA repair in mammalian liver cell cultures.

The genotoxic activity of four mycotoxins has been studied. A high level of somatic mutagenesis in imaginal disk of Drosophila melanogaster larvae and DNA repair synthesis in human embryo and adult rat liver cells cultures was induced only by the strong carcinogen aflatoxin B1. Patulin somewhat elevated the level of somatic mutations in D. melanogaster, but did not elicit DNA repair synthesis. Citrinin and stachybotryotoxin were inactive in both systems.

Aflatoxin B1↗

A new mycotoxin from Aspergillus candidus Link isolated from rough rice.

A new mycotoxin (AcT1) was obtained from the mycelium of Aspergillus candidus Link isolated from rough rice stored under tropical conditions. AcT1 with the mol-formula C28H40O showed bright greenish blue fluorescence under uv (254 nm). The spectral and other characteristics indicated that the compound was a new one. The LD50 of the toxin on white rats was found to be 4.5 mg/Kg when injected intraperitoneally.

Animals↗

Mycotoxin production on different cultivars and lines of broad bean (Vicia faba L.) seeds in Egypt.

One hundred different cultivars and lines of broad bean (Vicia faba L.) seed samples were inoculated with Aspergillus flavus Link (CMI 102135) to determine varietal differences which may support or resist aflatoxin production. Thin-layer chromatographic analysis of the chloroform extracts of the different seed samples revealed that 11 cultivars/lines were highly resistant to seed invasion and aflatoxin production while 9 cultivars/line showed partial resistance. The remaining 80 samples were susceptible to the establishment of A. flavus and aflatoxin accumulation. All the resistant cultivars/lines seed samples were inoculated also with three local isolates of fungi namely; Stachybotrys chartarum (Ehrenb. ex Link) Hughes, Aspergillus ochraceus Wilhelm, and Fusarium oxysporum Schlecht. The resistant seed samples were also resistant for colonization with these fungi and mycotoxin formation.

Aflatoxins↗

Toxicity of the mycotoxins fumonisins B1 and B2 and Alternaria alternata f. sp. lycopersici toxin (AAL) in cultured mammalian cells.

Fumonisins B1 and B2 and AAL toxin are a series of structurally related mycotoxins. Fumonisins B1 and B2, produced by Fusarium moniliforme Sheldon induce toxic hepatitis and hepatomas in rats and leukoencephalomalacia in horses. The cancer-promotion assay which has been used to guide their purification is slow and consumes large amounts of sample. We have examined a series of cultured mammalian cell lines in order to develop a more rapid and sensitive bioassay system, which may be useful for examining structure-activity relationships and the mechanism(s) of action of these toxins. Of 9 rat hepatoma cell lines tested, all except the two most de-differentiated line were sensitive to the three toxins, with a toxic response visible by 48 h. Approximate IC50 values for the most sensitive hepatoma line, H4TG, were 4, 2 and 10 micrograms/ml for fumonisins B1, B2 and AAL toxin, respectively, in 100 microliters cultures. Among 15 cell lines from other sources, only MDCK dog kidney epithelial cells were sensitive (IC50 = 2.5, 2 and 5 micrograms/ml, respectively). Studies in co-cultures of sensitive and insensitive cell lines and in cultures of a sensitive cell line over a range of cell densities indicated that cytotoxicity of fumonisins B1 and B2 does not involve metabolite activation to a derivative stable enough to diffuse to adjacent cells.

Alternaria↗

Tremorgenic mycotoxins produced by strains of Penicillium spp. isolated from toxic Poa huecu parodi.

Seventeen strains of Penicillium spp. have been isolated from Poa huecu Parodi from the Zapala zone, exhibiting toxicity to sheet. The following strains have been identified: P. crustosum, cyclopium, notatum, palitans, puberulum, verrucosum, viridicatum and Penicillium spp. The toxigenic capacity of the strains was studied after growing them under suitable conditions. Toxins produced were analysed by thin layer chromatography (TLC). Penitrem A (PA) and Penitrem B (PB) neurotoxins were identified and quantitated in twelve strains; verruculogen (VERR) and fumitremorgen B (FTB) being present in one of them. The effect of these mycotoxins was studied in mice. Neurological symptoms characteristic of the intoxication by tremorgenic toxins and similar to those observed in sheep suffering from 'huecu's disease' were observed. The possible role of these toxins as causative agents of 'huecu's disease' is discussed.

Animals↗

Cytotoxicity of Fusarium species mycotoxins and culture filtrates of Fusarium species isolated from the medicinal plant Tribulus terrestris to mammalian cells.

Ayurvedic medicine, which uses decoctions made of medicinal plants, is used to cure diseases in many Asian countries including Sri Lanka. Although proper storage facilities for medicinal plants are unavailable in Sri Lanka, neither the potential for growth of toxigenic fungi nor their ability to produce mycotoxins in stored medicinal plants has been investigated. We isolated three Fusarium species, F. culmorum, F. acuminatum and F. graminearum from the medicinal plant Tribulus terrestris. Culture extracts of the 3 Fusarium spp. were cytotoxic to mammalian cell lines BHK-21 and HEP-2. Three toxic metabolites produced by Fusarium spp; T-2 toxin, zearalenone, and diacetoxyscirpenol were also cytotoxic to the mammalian cell lines. The 3 Fusarium spp. grown on rice media produced zearalenone. Plant material destined for medicinal use should be stored under suitable conditions to prevent growth of naturally occurring toxigenic fungi prior to its use.

Animals↗

[Antibacterial and genotoxic properties of 33 mycotoxins].

Most of the 33 fungal metabolites tested provoke: Bacterial growth inhibition of Bacillus thuringiensis similar to lethal effect of antibiotics. Positive response in the 'Rec' assay using strains of Bacillus subtilis; this fact shows that these toxins are DNA modifying agents. Enlargement of cell volume in the first bacteria species; this cell-abnormality induction resembles those obtained with mitomycin C. Correlation between elongation of cells (filamentation) and in vivo carcinogenicity of mycotoxins is discussed. The filamentation should be an expression of a perturbated DNA replication (S.O.S.-error prone repair) as the consequence of DNA damages induced by genotoxic agents (i.e. carcinogens).

Bacillus subtilis↗

Influence of Fusarium and Myrothecium mycotoxins on dehydrogenase activity of Saccharomyces cerevisiae.

The Fusarium and Myrothecium mycotoxins roridin A, diacetoxyscirpenol, verrucarin A, T-2 toxin and zearalenone (10(-2) and 10(-3) mg/ml) inhibit the unspecific dehydrogenase activity of baker's yeast (Saccharomyces cerevisiae) in vivo. The action of these toxins is in the same order as that of aflatoxin B1. It is suggested that at least the trichothecenes decrease the dehydrogenase activity by an interaction with thiol groups of the active center of the enzymes.

Fusarium↗

Mycotoxins produced by Fusarium oxysporum in the seeds of Brassica campestris during storage.

Three recognized mycotoxins, viz. diaxetoxyscirpenol, T-2 toxin and zearalenone, along with the fatty esters of these toxins were isolated from the flask culture extractives of Fusarium oxysporum Schlecht (IMI 273220) as also from seeds of Brassica campestris var. sarson (mustard) infected with this fungus in storage. Evaluation of biological activity and toxicity of the extractives and the effects of prolonged ingestion of the mouldy seeds by animals suggest that the infected seeds may present high toxin-risk to humans.

Animals↗

Mycotoxins produced by toxic Fusarium isolates obtained from agricultural and nonagricultural areas (Arctic) of Norway.

Twenty-five isolates of F. acuminatum, 38 of F. avenaceum, 1 of F. culmorum, 31 of F. oxysporum and 56 of F. sambucinum were obtained in 1983, 1984 and 1986 from cereal grains and soil from various parts of Norway. The isolates were grown on an autoclaved Uncle Ben's parboiled rice medium and examined for production of trichothecenes and other toxins and for toxicity in rat feeding tests. F. culmorum N46C(2) and Fusarium sambucimum 45-86-A produced zearalenone (F-2) 864 and 665 ppm, respectively and caused uterine enlargement in rats. Most of these isolates produced no known trichothecene mycotoxins that could account for the toxicity that was demonstrated in the rat feeding tests. All but F. avenaceum N26B produced fusarin C (1.5 ppm) but caused no toxic effects in rat feeding test. None of the isolates produced fusarochromanone (TDP-1). Thirteen isolates of F. acuminatum, 16 of F. avenaceum, 14 of F. oxysporum and 3 of F. sambucinum produced a cytotoxic factor which we named HM-8. One isolate of F. avenaceum, 12 of F. oxysporum and 46 of F. sambucinum produced a hemorrhagic factor which we named H-1 (wortmannin). Twenty isolates of F. acuminatum, 22 of F. avenaceum, 17 of F. oxysporum and 1 of F. sambucinum produced moniliformin. Four isolates of F. acuminatum, 9 of F. avenaceum, 25 of F. oxysporum and 52 of F. sambucinum caused death to rats. Three isolates of F. avenaceum, 19 of F. oxysporum and 47 of F. sambucinum induced hemorrhage in various organs. All isolates caused decreased weight gain, relative to the control diets.

Amino Acids↗

Natural occurrence of the mycotoxin penitrem A in moldy cream cheese.

The occurrence of the mycotoxin penitrem A in refrigerated cream cheese that was involved in the intoxication of two dogs is described. Penitrem A and the fungus Penicillium crustosum were isolated from the cream cheese and identified. This is believed to be the first definitive case of the natural occurrence of penitrem A.

Animals↗

Mycotoxins producing fungi and mycoflora of air-dust from Taif, Saudi Arabia.

Using the dilution plate method, 70 species and 31 genera were collected from 20 dust samples on glucose (28 genera and 64 species) and cellulose Czapek's agar (22 genera and 46 species) at 28 degrees C. The most common fungi were Aspergillus niger, A. flavus, A. flavus var. columnaris, Phoma glomerata, Fusarium oxysporum, Penicillium chrysogenum and Mucor racemosus; and A. nidulans, Phoma humicola, Drechslera spicifera and Stachybotrys chartarum on the two media, respectively. Toxicity test showed that about 85% of the isolates tested were toxic to brine shrimp (Artemia salina). Thin layer chromatographic analysis revealed that 13 out of 23 toxic isolates produced known mycotoxins. Toxins identified were: aflatoxins B1 and B2, Kojic acid and trichodermine.

Aflatoxin B1↗

Mycotoxin formation in moist wheat under controlled temperatures.

One-kilogram parcels of wheat with 20.5% moisture content were maintained at 15 degrees and 22 degrees C for 10 weeks to study quality changes. Temperature, moisture, oxygen and carbon dioxide levels, microflora incidence and abundance, seed germination, fat acidity values, aflatoxins, sterigmatocystin, ochratoxin A, penicillic acid, citrinin and zearalenone were monitored. By two weeks, trace levels of ochratoxin had formed at both temperatures. By 10 weeks, the wheat contained at least three times more ochratoxin A at 22 degrees C than at 15 degrees C. Strains of Penicillium verrucosum var. cyclopium were associated with ochratoxin A production. No other mycotoxins were detected. The effect of temperature was significant for all variables (greater effect at 22 degree C) except A. glaucus gr. and Penicillium (P less than .01). The effect of time was significant for all variables except bacteria (P less than .01). The shape of the response was fully characterized by the linear and quadratic terms, except for % moisture which was linear only, and for bacteria for which time was not significant. The interaction between time and temperature was significant (P less than .01) for total fungal propagule count, % moisture, and Aspergillus versicolor, indicative of the steeper rise in slope for 22 degrees C.

Food Contamination↗

Fusarium species and their mycotoxins in infected corn in Italy.

Surveys of corn (infected plants and commercial kernels) for Fusarium species and their mycotoxins were carried out on samples collected all over Italy and from some European and mediterranean countries. Investigations on samples of corn stalk and ear rot standing in the field, mainly collected in southern Italy, proved to be contaminated with zearalenone (ZON), zearalenols (ZOL), and deoxynivalenol (DON). The Fusarium species most frequently isolated, and their recorded toxigenic capability (in parentheses), were: F. moniliforme; F. culmorum (ZON, ZOL, DON, 3AcDON); F. equiseti (ZON, ZOL); and F. proliferatum (MF). Along with these species, F. graminearum group 2 (ZON, DON and/or 3AcDON or 15AcDON); F. chlamydosporum; F. acuminatum (type-A trichothecene derivatives); and F. semitectum were often found to be associated. F. heterosporum (ZON, ZOL); F. solani; F. crookwellense (ZON, ZOL, FUS, NIV); F. oxysporum (MF); F. avenaceum (MF); F. sporotrichioides (T-2 toxin and derivatives); and F. poae (DAS, MAS) were occasionally isolated.

Fusarium↗

Genotoxic activity of some mycotoxins using the SOS chromotest.

The genotoxic activity of 11 mycotoxins was investigated in Escherichia coli K 12. The induction of the SOS function sfi A whose level of expression is monitored by means of a sfi A::lac Z operon fusion was assayed by measuring the beta-galactosidase activity in the PQ 37 strain. Most of these fungal metabolites did not induce SOS response in this bacterial test. Only aflatoxicol, a reduced metabolite of aflatoxin B1 was well detected as an SOS inducer if metabolic activation was performed. Patulin, penicillic acid and viomellein are only weak inducing agents. The other fungal compounds tested failed to demonstrate a positive SOS inducing activity. Relationship between SOS chromotest, mutagenicity to Salmonella typhimurium and in vivo carcinogenicity was discussed.

Colorimetry↗