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Effect of fungistatic agent 2-deoxy-D-glucose on mycotoxins from Penicillium expansum.

Mycotoxin production from Penicillium expansum was effected by the 'biocontrol enhancer' 2-deoxy-D-glucose (DOG). Citrinin and gentisyl alcohol were produced in higher amounts, whereas patulin was inhibited, which represents a differential effect on the polyketide metabolic pathway. These effects have implications for the use of DOG in biocontrol.

Antimetabolites↗

Mycotoxin-producing potential of mold flora of dried beans.

To evaluate the potential for mycotoxin production by molds in dried beans, the mold flora of 114 samples was determined both before and after surface disinfection of the beans with 5% NaOCl. Surface disinfection substantially reduced mold incidence, indicating that contamination was mainly on the surface. The flora, both before and after disinfection, was dominated by species of the Aspergillus glaucus group, the toxicogenic species A ochracues, Penicillium cyclopium, and P. viridicatum, and species of Alternaria, Cladosporium, and Fusarium. The toxicogenic species Aspergillus flavis, A. versicolor, Penicillium Citrinum, P. expansum, P. islandicum, and P. urticae were encountered less frequently. Of 209 species of Aspergillus and Penicillium screened for mycotoxin production on sterile rice substrate, 114 produced one or more of the following mycotoxins: A. flavus, aflatoxins; A. ochraceus, ochratoxins; A. nidulans, A. unguis, and A. versicolor, sterigmatocystin; P. cyclopium, penicillic acid; P. citrinum and P. viridicatum, citrinin; P. urticae, patulin and griseofulvin. Sterigmatocystin production by A. unguis is reported for the first time.

Aflatoxins↗

Mycotoxins.

Mycotoxins are secondary metabolites produced by microfungi that are capable of causing disease and death in humans and other animals. Because of their pharmacological activity, some mycotoxins or mycotoxin derivatives have found use as antibiotics, growth promotants, and other kinds of drugs; still others have been implicated as chemical warfare agents. This review focuses on the most important ones associated with human and veterinary diseases, including aflatoxin, citrinin, ergot akaloids, fumonisins, ochratoxin A, patulin, trichothecenes, and zearalenone.

Aflatoxins↗

Digestion of dry matter and amino acids and energy utilization by chicks fed molded corn containing mycotoxins.

A study was conducted to determine the ability of chicks to digest dry matter and amino acids and to utilize the energy from uncontaminated corn and contaminated corn containing mycotoxins. The contaminated corn contained either aflatoxin from natural molding, citrinin by artificially molding with a Penicillum spp., or ochratoxin by inoculation with Aspergillus ochraceus. Compared to uncontaminated corn, none of the criteria measured was affected in corn contaminated with aflatoxin. Dry matter digestibility, amino acid digestibility, and energy utilization were reduced (P less than .05) when corn was artificially molded with the Pencillium spp. and with A. ochraceus.

Aflatoxins↗

Survey of molds and mycotoxins for their ability to cause feed refusal in chickens.

The cause of feed refusal by poultry is an important problem that has not been established. T-2 toxin, a culture filtrate of Fusarium roseum NRRL 1181 containing a high concentration of diacetoxyscirpenol, and a filtrate of Penicillium citrinum containing an unknown factor produced refusal activity in week-old chickens. Citrinin stimulated water consumption while aflatoxin did not affect feed consumption. Detection of refusal activity was more sensitive when the refusal factor was presented in the drinking water than in the feed. The experimental model used here was sensitive, and, with appropriate controls, would appear suitable for the isolation and study of new factors capable of causing feed refusal in poultry.

Aflatoxins↗

Detection of twelve mycotoxins in mixed animal feedstuffs, using a novel membrane cleanup procedure.

A multimycotoxin thin layer chromatographic screening method is described which is applicable to most animal feedstuffs. Interference from nonspecific lipid, pigment, and other components of simple and mixed feeds is reduced to a minimum by using a membrane cleanup step. Aflatoxins B1, B2, G1, and G2, citrinin, diacetoxyscirpenol, ochratoxin A, patulin, penitrem A, sterigmatocystin, T-2 toxin, and zearalenone may be reliably detected. The sensitivity of the method is generally low for mixed feeds but even so aflatoxin B1 can be detected at a level of 3 ppb and ochratoxin A at 80 ppb. While the basic method is less sensitive for sterigmatocystin (330 ppb), patulin (600 ppb), zearalenone (1000 ppb), and the trichothecenes (1000-4000 ppb), it may be adapted so as to reduce the above detection limits when the presence of these toxins is suspected. Lower levels may be detected in extracts of simple feeds.

Aflatoxins↗

Genotoxicity of ochratoxin A and structurally related compounds in Escherichia coli strains: studies on their mode of action.

Ochratoxin A, ochratoxin alpha (its major metabolite in rodents) and seven structurally related substances were assayed for SOS DNA repair inducing activity in Escherichia coli PQ37 strain. At a concentration range of 0.1-4 mM, ochratoxin A, chloroxine, 5-chloro-8-quinolinol, 4-chloro-meta-cresol and chloroxylenol were found to induce SOS-DNA repair in the absence of an exogenous metabolic activation system. Ochratoxin B, ochratoxin alpha, 5-chlorosalicylic acid and citrinin were inactive, but all except ochratoxin alpha were cytotoxic. Thus, the presence of a chlorine at C-5 in ochratoxin A and in other analogues appears to be one determinant of their genotoxicity. In order to ascertain whether this reactivity involves a bacterial glutathione conjugation reaction, we investigated the modifying effect on the genotoxicity of ochratoxin A of amino oxyacetic acid, an inhibitor of cysteine conjugate beta-lyase. Amino oxyacetic acid decreased the cytotoxicity of ochratoxin A but did not alter its genotoxic activity, suggesting the formation of a cytotoxic thiol-containing derivative. The way in which ochratoxin A and some of its active analogues induce SOS DNA repair activity was further investigated in E. coli PQ37 and in three derived strains (PQ300, OG100 and OG400, containing deletions within the oxy R regulon). The response in PQ37 strain was measured in the absence and presence of Trolox C, a hydrosoluble form of vitamin E. Trolox C completely quenched the genotoxicity of ochratoxin A, which was no greater in mutated than in wild type strains. These results implicate an ochratoxin A-derived free radical rather than reduced oxygen species as genotoxic intermediate(s) in bacteria.

Aminooxyacetic Acid↗

[Induction by mycotoxins of somatic mosaicism in Drosophila and DNA repair in mammalian liver cell cultures].

The genotoxic activity of four mycotoxins has been studied. High level of somatic mutagenesis in imaginal discs of Drosophila melanogaster larvae and DNA repair synthesis in human embryo and adult rat liver cell cultures were inducible only by highly carcinogenic aflatoxin B1. Patulin, a weak direct-action carcinogenic substance, slightly elevated the mutagenesis in somatic cells of Drosophila but did not induce DNA repair synthesis in liver cell cultures. Citrinin that did not exhibit any carcinogenic properties when used alone and stachybotrotoxin with non-reported carcinogenic activity appeared inactive in the test-systems applied. The possibilities of rapid recognition of carcinogenic mycotoxins by detecting their genotoxic properties are discussed.

Aflatoxin B1↗

Mycotoxins transmitted into beer from contaminated grains during brewing.

Studies with aflatoxin B1, ochratoxin A, zearalenone, deoxynivalenol, and fumonisins B1 and B2 added at various stages of the brewing process show that these mycotoxins (or metabolites) may be transmitted from contaminated grains into beer. Citrinin does not survive the mashing step. Mycotoxins in beer could originate from the malted grain or from adjuncts. Although high incidences and concentrations of aflatoxins and zearalenone have been found in local beers brewed in Africa, aflatoxins have not been detected in European beers. Zearalenone and alpha- or beta-zearalenol (the metabolite likely to occur) have not been found in Canadian and European beers, except for one sample analyzed by thin-layer chromatography only. Ochratoxin A rarely has been detected at > 1 ng/mL in beer; liquid chromatographic methods with a 0.05-0.1 ng/mL detection limit, however, have shown moderately high incidences of trace levels. Deoxynivalenol, which survives the brewing process, has been found with high incidence in Canadian and European beers, with concentration of > 200 ng/mL reported in several German beers. Fumonisins B1 and B2 occur to a limited extent in beer.

Aflatoxin B1↗

Mycotoxins in cereal grain. Part IV. Inactivation of ochratoxin A and other mycotoxins during ammoniation.

Addition of ammonia to final concentration 2% inactivates ochratoxin A, aflatoxin, citrinin, penicillic acid and partially zearalenon at temperature 20-50 degrees C. Detoxification of contaminated cereal grain (wheat, corn or barley) can be performed on a farm using ammoniation without special investment during 4 to 6 weeks. Ammoniation changes nutritional value of grain as feed in a small extent.

Ammonia↗

[Carcinogenic and genotoxic effects of mycotoxins].

Aflatoxin B1 is the most potent hepatocarcinogen among the naturally occurring compounds. It's carcinogenic effect was confirmed by longterm-studies in several animal species. There exists detailed knowledge on the molecular mechanism of the carcinogenic effect caused by aflatoxin B1. At present there does not exist a reliable evidence that human primary hepatocellular carcinoma is caused by aflatoxins. Sterigmatocystin is a hepatocarcinogen too. The genotoxicity of sterigmatocystin likewise as of aflatoxin B1 was detected by short-term-tests with microbial systems and mammalian cells in vitro. It is disputable whether ochratoxin A, citrinin and T-2 toxin are carcinogenic or not. Diacetoxyscirpenol, zearalenone and patulin are estimated as not carcinogenic after longterm-studies with oral toxin administration. Patulin and penicillic acid are genotoxic in microbial systems respectively in mammalian cells in vitro. Luteoskyrin is carcinogenic, but there was not detected any genotoxicity.

Animals↗

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↗

Genotoxicity of fungi evaluated by SOS microplate assay.

By an introduction of sodium dodecylsulfate for cell lysis and immunomicroplate for mass assay, the modified SOS microplate assay method was established and applied for the evaluation of genotoxicity of mycotoxins and fungal cultures. Among 20 mycotoxins, the carcinogenic dihydrobisfuranoids such as aflatoxin B1, sterigmatocystin, and versicolorin A were positive in the presence of the activation system. While, the carcinogenic anthraquinones and lactones such as luteoskyrin, rugulosin, ochratoxin A, patulin, and citrinin were negative. The survey on genotoxic fungi revealed that, among 15 fungal isolates Aspergillus versicolor, Emericella acristata, and others were positive. Additional survey on 265 fungal isolates have revealed that various Aspergillus genera such as A. flavus, A. parasiticus, A. ustus, A. nidulans, and others were positive for SOS induction, along with several isolates of Fusarium moniliforme. The chemical analysis revealed that the dihydrobisfuranoids such as aflatoxin B1, and sterigmatocystin were the major genotoxic metabolites of several Aspergillus species. The SOS microplate assay system is a simple and rapid procedure for the mass screening of genotoxic fungi, particularly of the dihydrobisfuranoids-producing strains.

Animals↗

Induction of apoptosis by T-2 toxin and other natural toxins in HL-60 human promyelotic leukemia cells.

Based on the DNA fragmentation profile in gel electrophoresis and the morphological changes in electron microscopy, the induction of apoptotic nuclear changes by mycotoxins and other microbial products, in total 31 chemicals, was investigated in HL-60 human promyelotic leukemia cells, along with the cytotoxicity tests with 3-[4,5-dimethylthiazol-zyl]-2,5-diphenyltetrazolium bromide (MTT) and trypan blue exclusion. Among the chemicals tested, trichothecenes (T-2 toxin, roridin A, nivalenol, deoxynivalenol), certain anthraquinones (luteoskyrin, skyrin, 2-hydroxyemodin), diketopiperazines (emethallicin A, emestrin), isocoumarins (ochratoxin A, citrinin), lactone (penicillic acid), dihydrobisfuran (aflatoxin B1), potassium ionophore (valinomycin), and an inhibitor of interleukin-2 synthesis (cyclosporin A) were positive for the induction of DNA fragmentation. No DNA fragmentation was observed under the present conditions with fumonisin B1, cyclic peptides (cyclochlorotine, phalloidin, microcystin-LR), certain anthraquinones (emodin, chrysophanol, rugulosin), and others (sterigmatocystin, cytochalasin A, griseofulvin, fusaric acid, kojic acid, rubratoxin B, butenolide, wortmannin, FK506, and sphingosine). The apoptotic changes in the cells exposed to T-2 toxin and luteoskyrin were confirmed by electron microscopic observation. Detailed experiments on dose and time dependencies revealed that T-2 toxin induced the apoptosis at 10 ng/ml (= 4 x 10(-8) M) levels within 2-6 hr without significant cytotoxicity evaluated by the dye exclusion and MTT.

Anthraquinones↗

Biology and ecology of mycotoxigenic Aspergillus species as related to economic and health concerns.

The fungal genus Aspergillus was established in 1729, and includes species that are adapted to a wide range of environmental conditions. Many aspergilli produce mycotoxins in foods that may be toxic, mutagenic or carcinogenic in animals. Most of the Aspergillus species are soil fungi or saprophytes but some are capable of causing decay in storage, disease in plants or invasive disease in humans and animals. Major agricultural commodities affected before or after harvest by fungal growth and mycotoxins include corn, peanuts, cottonseed, rice, tree nuts, cereal grains, and fruits. Animal products (meat, milk and eggs) can become contaminated because of diet. Aspergillus flavus, A. parasiticus, A. ochraceus, A. niger, A. fumigatus and other aspergilli produce mycotoxins of concern. These include the aflatoxins and ochratoxins, as well as cyclopiazonic acid, patulin, sterigmatocystin, gliotoxin, citrinin and other potentially toxic metabolites.

Animals↗

Ochratoxin A production in Bavarian cereal grains stored at 15 and 19% moisture content.

Wheat, barley, and maize, each in 15-kg parcels at 15 and 19% initial moisture content (IMC), were kept in a Bavarian farm granary from June through November 1990. During this period, the grain at each IMC was analyzed for mycotoxins and monitored for grain temperature, carbon dioxide, seed germination, and microfloral incidence and abundance. Barley and maize stored for 20 weeks at 19% IMC contained ochratoxin A in amounts of 70 and 90 microg/kg, respectively. This mycotoxin was not detected in wheat stored at 19% IMC, nor in the grains stored at 15% IMC. Aflatoxin B1, sterigmatocystin, citrinin, and zearalenone were also assayed but not detected in grains stored at either IMC. Principal component analysis of the data indicated that ochratoxin A was produced in a damp niche in maize, when abundant metabolic activity and CO2 production by Penicillium glandicola and Aspergillus spp. were common.

Edible Grain↗

Mycotoxins and mycotoxigenic moulds in nuts and sunflower seeds for human consumption.

A survey was carried out to obtain data on the occurrence of mycotoxins and the mycotoxin-producing potential of fungi isolated from nuts (almonds, peanuts, hazelnuts, pistachio nuts) and sunflower seeds in Spain. Thin-layer chromatography was used to separate the toxins. Aflatoxins were detected in one sample of almonds (95 ppb aflatoxin B1 and 15 ppb aflatoxin B2) and in one sample of peanuts at a level below 10 ppb of aflatoxin B1. 100% of samples showed variable incidence of fungal contamination. The predominant fungi present in samples were Penicillium spp, Aspergillus niger, A. flavus, A. glaucus and Rhizopus spp. The results showed that isolates of different species were able to produce aflatoxins B1, B2, G1, and G2, sterigmatocystin, ochratoxin A, patulin, citrinin, penicillic acid, zearalenone, and griseofulvin.

Analysis of Variance↗

Mycotoxin formation in moist 2-row and 6-row barley during granary storage.

Eleven-kilogram parcels of 2-row and 6-row barley initially at 18% moisture content were implanted in dry bulk oats in a farm granary in Manitoba for 60 weeks between August 1983 and October 1984. Temperature, moisture content, O2 and CO2 levels, fat acidity values, seed germination, microfloral incidence and abundance and the presence of major mycotoxins (aflatoxins, sterigmatocystin, ochratoxin A, citrinin, penicillinic acid, patulin) were monitored. Ochratoxin A reached maximum levels of 0.97 ppm by week 24 in the 6-row barley, and 0.05 ppm by week 28 in the 2-row; no other mycotoxins were detected. The effect of cultivar type was significant (P less than 0.01) with greater effects in the 6-row barley for the following parameters: fat acidity value, germination, incidence of Alternaria alternata, Penicillium spp. and Helminthosporium sativum, total fungal propagule count and ochratoxin A levels. The effect of time was significant (P less than 0.05) for all variables except oxygen, carbon dioxide, Aspergillus versicolor, and total fungal propagule count. The interaction between cultivar and time was significant (P less than 0.01) for Alternaria and Helminthosporium only.

Alternaria↗