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PubMed1990

Mycotoxin formation in HY-320 wheat during granary storage at 15 and 19% moisture content.

Eleven-kilogram parcels of HY-320 wheat, a cultivar of the new Canada Prairie Spring class, were kept at 15 and 19% initial moisture contents (IMC) in simulated storage in a Manitoba farm granary for 60 weeks to determine biotic and abiotic changes and mycotoxin production. Ochratoxin A reached a maximum of 0.24 ppm by week 20 in the 19% IMC wheat, but was absent in the 15% IMC wheat; no other mycotoxins were detected. Temperature, moisture content, O2 and CO2 levels, fat acidity values, seed germination, microfloral incidence and abundance, and the presence of other mycotoxins were monitored. Principal component analysis of all variables showed that the first principal components accounted for 32-41% of the system variability, and contained the ochratoxin A variable. Ochratoxin A was produced in moist grain that had decreased seed germination and Alternaria activity, and high fungal activity by Penicillium and Aspergillus versicolor. Compared to other stored cereals previously studied, HY-320 wheat would be ranked in a low-risk category for mycotoxin formation, based on the ochratoxin A levels observed.

Carbon Dioxide↗

PubMed2005

Fungal populations and mycotoxins in silage in Assiut and Sohag governorates in Egypt, with a special reference to characteristic Aspergilli toxins.

Forty silage samples were collected from Assiut and Sohag governorates in Egypt to measure the presence of fungal population in silage. Forty-three species and 2 species varieties belonging to 17 genera were isolated using glucose Czapek's and Sabouraud's dextrose agar media at 28 degrees C. The most prevalent genera were Aspergillus (57.5 and 100 of the samples), Penicillium (100 and 55%) on the two mentioned media, respectively. Also, Fusarium oxysporum and Gibberella fujikurori were recovered in moderate incidences. Mycotoxin profiles were also determined in these samples: Aflatoxins showed the highest incidence rates of occurrence, it occurred in 22.5% of all samples analyzed. Other mycotoxins were detected from all samples (T2 toxins and sterigmatocystin at incidence of 7.5 and 5%, respectively). The screening of the characteristics mycotoxins of different isolates of Aspergillus isolated from silage samples was tested. The results clarified that some mycotoxins (aflatoxins-aspergillic acid-beta nitro propionic acid-cyclopiazonic acid-kojic acid and sterigmatocystin) were produced by some isolates of A. flavus. Some isolates of A.fumigatus could produce gliotoxin and verrucologen. All of A. niger isolates tested were able to produce kojic acid. One isolate of A. ochraceous formed ochratoxin A and other isolate produced penicillic acid. Concerning A. terreus isolates, the results showed that 5 isolates were able to produce citrinin and 4 isolates had ability to produce patulin. A. versicolor isolates showed the ability to produce ochratoxin A.

Animals↗

PubMed2006

Mycotoxin contamination of food in Europe: early detection and prevention strategies.

This paper reviews the early detection and prevention strategies which have been employed in Europe for the control of mycotoxin contamination of food in the context of a hazard analysis critical control point (HACCP) framework. The critical control points (CCPs) in the whole food chain where mycotoxins such as trichothecenes and ochratoxins are important have been identified. Ecological studies on the effect of environmental factors which are marginal for growth and mycotoxin production have been identified for Fusarium culmorum and F. graminearum (deoxynivlenol production), and for Penicillium verrucosum and Aspergillus ochraceus (ochratoxin production) in relation to cereal production and for A. carbonarius in relation to grapes and wine production (ochratoxin formation). To minimise the entry of these mycotoxins into the food chain, effective and rapid diagnostic tools are required to monitor the CCPs effectively. To this end the potential use of molecular imprinted polymers, lateral flow devices and molecular-based techniques for the rapid detection and quantification of the mycotoxigenic moulds or their toxins have also been developed.

Aspergillus ochraceus↗

PubMed2006

Mycotoxins in Mexico: epidemiology, management, and control strategies.

Mycotoxin-producing molds species are extremely common. Many of them provoke serious diseases in humans and animals. These toxins occur, with varying severity, in agricultural products. Since Mexico is a big crops producer, the importance of the presence of these toxins is high. Currently, the Mexican regulation establishes limits only for aflatoxins in cereals and cereal products. No limits are set for other mycotoxins. Epidemiological data although limited, has shown that an important number of samples contain mycotoxin limits above those established abroad. Several strategies for reducing contamination have been conducted in the country such as development of hybrid corn, control of insect population, use of natural products, and modification of nixtamalization and extraction procedures. Although significant research on this field has been conducted, there is still a great need for information to determine the incidence of mycotoxins in Mexican products.

Aflatoxins↗

PubMed2005

A model transgenic cereal plant with detoxification activity for the estrogenic mycotoxin zearalenone.

Zearalenone (ZEN) is an estrogenic mycotoxin produced by the necrotrophic cereal pathogen Fusarium graminearum. This mycotoxin is detoxified by ZHD101, a lactonohydrolase from Clonostachys rosea, or EGFP:ZHD101, its fusion to the C-terminus of an enhanced green fluorescence protein. We previously showed that egfp:zhd101 is efficiently expressed in T(0) leaves of rice. In this study, we assessed the feasibility of in planta detoxification of the mycotoxin using progeny. When protein extract from T(1) leaves was incubated with ZEN, the amount of the toxin decreased significantly as measured by HPLC. ZEN degradation activity was also detected in vivo in transgenic T(2) seeds. These results suggest that zhd101 can be exploited as an efficient and cost-effective system for protection of important cereals that are more susceptible to the pathogen (e.g., wheat and maize) from contamination with the estrogenic mycotoxin.

Base Sequence↗

PubMedDate not supplied

Mycotoxins: food contamination, mechanism, carcinogenic potential and preventive measures.

Mycotoxins constitute a large number of naturally occurring fungal secondary metabolites with very diversified toxic effects in humans and animals. Among many mycotoxins discovered, aflatoxins, ochratoxin A, sterigmatocystin and several others are identified as carcinogens; several others were found to be mutagenic. Nevertheless, aflatoxin B1 has been found to be one of the most potent carcinogens and contamination of aflatoxins in the food supply is still a major concern. Whereas extensive studies have been made on aflatoxins, little is known about the mode of action of other carcinogenic and mutagenic mycotoxins. Recent progress on research for the carcinogenic and mutagenic mycotoxins is presented in this review with emphasis on their contamination in foods, their carcinogenic potential to humans, and the mode of action as well as possible preventive measures.

Aflatoxins↗

PubMed1988

Mould spoilage and mycotoxin formation in grains as controlled by physical means.

Low oxygen concentrations (less than 1%) and/or increased concentrations of CO2 or N2 have been found to be highly effective in preventing the development of mould on grain and in inhibiting selected mycotoxins, e.g. aflatoxins, ochratoxin, patulin, penicillic acid and T-2. However, the levels of CO2 needed to inhibit mould growth are much higher than those required for the inhibition of mycotoxin production. The degree of inhibition achieved by elevated CO2 concentrations is dependent on other environmental factors, such as relative humidity (RH) and temperature. Nevertheless, the biosynthetic pathways for mycotoxin production are merely blocked, but not damaged by high CO2 levels. Irradiation has been shown to destroy the conidia of moulds but the information concerning the effect of irradiation on mycotoxin formation seems to be contradictory. Aflatoxin production was increased in irradiated wheat grain, but decreased in barley and maize when the grain was irradiated prior to inoculation. The number of spores in the inoculum, grain condition, relative humidity and other environmental factors could all affect the results obtained. However, ochratoxin formation by Aspergillus ochraceus was consistently enhanced by irradiation of spores or mycelium.

Edible Grain↗

PubMed1994

Direct detection using the Drosophila DNA-repair test and isolation of a DNA-damaging mycotoxin, 5,6-dihydropenicillic acid, in fungal culture.

The Drosophila DNA-repair test was used in an attempt to detect fungal production of DNA-damaging mycotoxins without an extraction process. 29 species of fungi, 13 Aspergillus, 12 Penicillium and four Fusarium were inoculated directly to a Drosophila medium, and the larvae were then bred in the mouldy medium. Production of DNA-damaging mycotoxin was detected directly by counting the decrease in the survival of DNA-repair-deficient flies. With the direct detection method, Aspergillus ochraceus, A. parasiticus and A. versicolor produced DNA-damaging mycotoxins. The same results were obtained with the mouldy medium extract using the standard DNA-repair test. The direct detection method was convenient for surveying the fungal production of DNA-damaging mycotoxins. The extracts of A. parasiticus and A. versicolor contained aflatoxin B1 and sterigmatocystin, respectively. The DNA-damaging compound in the extract of A. ochraceus was isolated and purified to clear, colourless 'needles'. With nuclear magnetic resonance-mass spectroscopy spectra, the compound was confirmed to be 5,6-dihydropenicillic acid, the DNA-damaging potency of which has not been previously reported.

Animals↗

PubMed2006

What the primary care pediatrician should know about syndromes associated with exposures to mycotoxins.

Disease associated with exposure to mycotoxins is known as the "Great Masquerader" of the 21st century because of its complex natural history involving different tissues and resembling different diseases at each stage in its evolution. It can present with a variety of nonspecific clinical signs and symptoms such as rash, conjunctivitis, epistaxis, apnea, cough, wheezing, nausea, and vomiting. Some cases of vomiting illness, bone marrow failure, acute pulmonary hemorrhage, and recurrent apnea and/or "pneumonia" are associated with exposure to mycotoxins. Familiarity with the symptoms of exposure to the major classes of mycotoxins enables the clinician to ask pertinent questions about possible fungal exposures and to remove the infant or child from the source of exposure, which could be contaminated food(s), clothing and furniture, or the indoor air of the home. Failure to prevent recurrent exposure often results in recurrent illness. A variety of other conditions, including hepatocellular and esophageal cancer and neural tube defects, are associated with consumption of foods contaminated with mycotoxins. Awareness of the short- and long-term consequences of exposures to these natural toxins helps pediatricians to serve as better advocates for children and families.

Acute Disease↗

PubMed2005

Cytotoxicity of Fusarium mycotoxins to mammalian cell cultures as determined by the MTT bioassay.

Fusarium mycotoxins occur worldwide in cereal grains and animal feeds and cause outbreaks of Fusarium mycotoxicoses in humans and animals. In this study mammalian cell cultures were used to screen the cytotoxicity of the most common Fusarium mycotoxins; deoxynivalenol (DON), zearalenone (ZEN), fumonisin B(1) (FB(1)) and moniliformin (MON). The most sensitive cell line for each Fusarium mycotoxin was determined for further toxicological investigations as an alternative to whole animal testing. Chinese hamster ovary cells (CHO-K1) were found to be the most sensitive for DON and FB(1) with IC(50) values of 0.27 and 85.5 microg/ml, respectively, after 48-h exposure. The hepatocellular carcinoma cells (HepG2) showed the highest sensitivity to MON with IC(50) values of 39.5 for 48 h and 26.8 microg/ml for 72-h exposure. Balb/c mice keratinocyte cell line (C5-O) was found to be the most sensitive to ZEN with IC(50) of 24.1 microg/ml after 72-h exposure. DON was found the most cytotoxic to the cell cultures of all the mycotoxins tested, followed by MON, ZEN, and FB(1). The results indicated that CHO-K1, C5-O, and HepG2 cells were found to be the sensitive cell lines for preliminary screening of DON, ZEN and MON contaminated feed and food extracts, respectively.

Animal Feed↗

PubMed2005

Fungal mycoflora and mycotoxins in Korean polished rice destined for humans.

Rice samples collected from the Republic of Korea were analyzed for fungal mycoflora and mycotoxins: fumonisins, ochratoxin A, trichothecenes, and zearalenone. The potential of the fungi to produce each mycotoxin was also examined, so that the fungal isolates associated with mycotoxins occurring in rice could be verified. Penicillium citrinum and Aspergillus candidus were the most prevalent species infecting the samples, while Fusarium proliferatum was found as the dominant Fusarium species. Ochratoxin A was the most commonly detected mycotoxin analyzed in the present study; moreover, its level in some samples was above the EU tolerable limit (3 ng/g). According to rice culture experiments, it was revealed that in Korea, fumonisins detected in rice were due to F. proliferatum infection, whereas the occurrence of ochratoxin A was caused by Penicillium verrucosum, though there were no symptoms of disease in rice found in any sample. Furthermore, there appears to be an uneven geographical distribution of P. verrucosum as well as ochratoxin A in that most of them are found in the rice samples produced in the northern region of Korea.

Food Contamination↗

PubMed2005

Mycotoxin-producing and other fungi isolated from grapes for wine production, with particular emphasis on ochratoxin A.

Mycotoxins are toxic secondary metabolites produced by filamentous fungi that have been detected in food commodities, including grapes and wine. A survey was conducted to assess mycotoxin-producing fungi in grapes destined for wine production. The mycotoxigenic capacity of the isolates was tested in culture media. Grapes were analyzed by plating methods from 4 Portuguese wine-growing regions at 3 maturation stages (pea berry, early veraison and ripe berry) between 2001 and 2003. From the 10,602 strains detected and identified, the most frequent genera were Cladosporium (25%), Alternaria (24%), Botrytis (15%), Penicillium (9%) and Aspergillus (8%). Most (92.0%) were non-mycotoxigenic or produced mycotoxins of unknown relevance to health. Potential producers of ochratoxin A (OTA) (Aspergillus niger aggregate, 5.4%, Aspergillus carbonarius, 0.6%) and trichothecenes (Fusarium spp., 0.4%; Trichothecium roseum, 0.8%) were the most frequent mycotoxigenic species isolated from grapes. OTA was detected in all cultures of A. carbonarius and 4% of A. niger aggregate strains. There was potential for OTA and trichothecene production in grapes by A. carbonarius and T. roseum, respectively, prior to harvest time. Data presented herein indicate that A. carbonarius is most likely to occur in vineyards with Mediterranean climates, while T. roseum is more likely to occur in more temperate climates, and is associated with gray rot. The present work emphasizes the need to use grapes under good condition so as to reduce the risk of contamination with mycotoxigenic fungi and subsequent mycotoxin occurrence in wine.

Aspergillus↗

PubMed2006

Effects of combinations of Fusarium mycotoxins on the inhibition of macromolecular synthesis, malondialdehyde levels, DNA methylation and fragmentation, and viability in Caco-2 cells.

We studied the interactive effects of either binary or tertiary mixtures of Fusarium mycotoxins, deoxynivalenol (DON), zearalenone (ZEA), and fumonisin B1 (FB1) on the human intestinal cell line, Caco-2, using the endpoints including malonedialdehyde (MDA) production, inhibition of protein and DNA syntheses, DNA methylation, DNA fragmentation, and cell viability as measured by the neutral red (NR) test. The mixtures of mycotoxins reduce cellular viability in increasing order: [FB1+ZEA]<[FB1+DON]<[ZEA+DON]<[FB1+DON+ZEA] in NR test. Because FB1 antagonizes the effects of estrogenic Zearalenone, FB1 was assayed against estradiol. In NR assay, mixture of FB1 and estradiol and/or ZEA improves Caco-2 cells viability in contrast to individual effects. Mixtures of ZEA or FB1 and DON, display synergistic effects in lipid peroxidation. The ability of the toxins to inhibit DNA synthesis is 45%, 70%, and 43% for 10 microM of ZEA, DON, and FBI, respectively. Their binary mixtures (at 10 microM each), inhibit DNA synthesis by 35%, 62%, and 65%, far less than additive effects. Surprisingly, the tertiary mixture (10 microM each) only inhibits DNA synthesis by 25%. ZEA, DON, and FB1 induce DNA fragmentation individually. However, mixtures of these mycotoxins always damage DNA to a greater extent. Each individual mycotoxin (10 microM) raises the percentage of 5-methylcytosine (m5dC) in DNA from 4.5% to 9%, while the combination does not increase this rate any further. Altogether, the data indicate that mixtures of Fusarium toxins are able to induce lipid peroxidation, DNA damage, DNA fragmentation, DNA methylation, and cytotoxicity in Caco-2 cells, and suggest a potential promoter effect in human intestinal cells.

Apoptosis↗

PubMed2006

Mycotoxin-induced depletion of intracellular glutathione and altered cytokine production in the human alveolar epithelial cell line A549.

Mould exposure has been associated with asthma and other inflammatory airway conditions. However, cellular effects of inhaled mould components are not well understood. We hypothesised that host defence mechanisms, such as production of cytokines (TGFbeta1, IL-6 and IL-8) and the intracellular antioxidant glutathione (GSH), could be adversely affected by different concentrations of mycotoxins. We studied the effects of citrinin and gliotoxin on lipopolysaccharide (LPS)-stimulated alveolar epithelial cells (A549). Cytokines in cell culture supernatants were analysed by ELISA and levels of GSH were measured by colorimetric (absorbance) determination. We found that GSH decreased in a dose- and time-dependent manner when cells were exposed to citrinin in particular. TGFbeta1 was moderately reduced at low mycotoxin concentrations but elevated at higher sub-toxic concentrations. A tendency for an inverse relationship between TGFbeta1 and GSH levels was observed. IL-6 and IL-8 were not significantly reduced at non-toxic mycotoxin concentrations. Thus, reduced epithelial GSH and TGFbeta1 levels combined with elevated IL-6 and IL-8 levels may result in increased pro-inflammatory activity during mycotoxin exposure. We suggest that this mechanism can contribute to inflammation in mould exposure.

Cell Line↗

PubMed2004

Production of toxic metabolites in Aspergillus niger, Aspergillus oryzae, and Trichoderma reesei: justification of mycotoxin testing in food grade enzyme preparations derived from the three fungi.

Aspergillus niger, Aspergillus oryzae, and Trichoderma reesei are three important production organisms used in industrial fermentations. Several of the fungal secondary metabolites produced by selected strains of these three fungi are capable of eliciting toxicity in animals. Among those toxic substances are the well-known mycotoxins 3-nitropropionic acid and ochratoxin A. However, many others, such as kojic acid, may not be true mycotoxins. The production, extraction, chemical structure, and the toxicity (expressed as LD(50)) of these substances are reviewed. Production of toxic secondary metabolites in A. niger, A. oryzae, and T. reesei is strain-specific and environment-dependent. Considering all of the safety measures taken in the industrial production process, these three fungal species are safe to use. The recently revised JECFA specification for mycotoxins in food enzyme preparations is also discussed. The extent of mycotoxin tests in food enzyme preparations should be judged on a case-by-case basis, through a careful evaluation based on knowledge of taxonomy, biochemistry, and genetics. In many cases, the testing scope at the level of genus should be sufficient. In other cases, the scope can even be further narrowed based on scientific knowledge and assessment.

Animals↗

PubMed2002

Determination of Penicillium mycotoxins in foods and feeds using liquid chromatography-mass spectrometry.

New LC-MS (full scan) and LC-MS-MS (selected ion reaction monitoring) methods for the simultaneous determination of mycophenolic acid, griseofulvin, roquefortine C, chaetoglobosin B, verruculogen and penitrem A, and other Penicillium derived mycotoxins in food and feed samples are described. The methodologies involve sample extraction with acetonitrile-water, defatting with hexane and quantification using LC-MS with atmospheric pressure chemical ionisation or LC-MS-MS. Detector responses, for each of the methods and mycotoxins, were found to be linear over the range 10-1000 ng of mycotoxin/g of extracted food mixture material. The mean recoveries (n = 3 to 6) of the mycotoxins from spiked food mixture samples determined using MS and MS-MS detection were 87-116 and 91-112%, respectively, for mycophenolic acid, 104-109 and 91-112%, respectively, for griseofulvin, 70-85 and 75-110%, respectively, for roquefortine C, 94-109 and 81-116%, respectively, for chaetoglobosin B, 110-115 and 90-106%, respectively, for verruculogen and 78-97 and 99-108%, respectively, for penitrem A. RSDs varied from 5.6% at the 1000 ng/g level to 23.1% at the 10 ng/g level. The limits of detection for the mycotoxins using MS and MS-MS were 70 and 10 ng/g, respectively, for mycophenolic acid, 10 and 5 ng/g, respectivley, for griseofulvin, 50 and 20 ng/g, respectively, for roquefortine C, 25 and 20 ng/g, respectively, for chaetoglobosin B, 25 and 20 ng/g, respectively, for verruculogen and 10 and 5 ng/g, respectively, for penitrem A.

Animal Feed↗

PubMed1997

Oxidative degradation and detoxification of mycotoxins using a novel source of ozone.

Practical methods to degrade mycotoxins using ozone gas (O3) have been limited due to low O3 production capabilities of conventional systems and their associated costs. Recent advances in electrochemistry (i.e. proton-exchange membrane and electrolysis technologies) have made available a novel and continuous source of O3 gas up to 20% by weight. It is possible that the rapid delivery of high concentrations of O3 will result in mycotoxin degradation in contaminated grains--with minimal destruction of nutrients. The major objectives of this study were to investigate the degradation and detoxification of common mycotoxins in the presence of high concentrations of O3. In this study, aqueous equimolar (32 microM) solutions of aflatoxins B1 (AfB1), B2 (AfB2), G1 (AfG1), G2 (AfG2), cyclopiazonic acid (CPA), fumonisin B1 (FB1), ochratoxin A (OA), patulin, secalonic acid D (SAD) and zearalenone (ZEN) were treated with 2, 10 and/or 20 weight% O3 over a period of 5.0 min and analysed by HPLC. Results indicated that AfB1 and AfG1 were rapidly degraded using 2% O3, while AfB2 and AfG2 were more resistant to oxidation and required higher levels of O3 (20%) for rapid degradation. In other studies, patulin, CPA, OA, SAD and ZEN were degraded at 15 sec, with no by-products detectable by HPLC. Additionally, the toxicity of these compounds (measured by a mycotoxin-sensitive bioassay) was significantly decreased following treatment with O3 for 15 sec. In another study, FB1 (following reaction with O3) was rapidly degraded at 15 sec, with the formation of new products. One of these appeared to be a 3-keto derivative of FB1. Importantly, degradation of FB1 did not correlate with detoxification, since FB1 solutions treated with O3 were still positive in two bioassay systems.

Aflatoxins↗

PubMed2001

Mycotoxin detoxication of animal feed by different adsorbents.

The contamination of animal feed with mycotoxins represents a worldwide problem for farmers. These toxins originate from molds whose growth on living and stored plants is almost unavoidable particularly under moist conditions. Mycotoxin-containing feed can cause serious diseases in farm animals resulting in suffering and even death and thus can cause substantial economic losses. The most applied method for protecting animals against mycotoxicosis is the utilization of adsorbents mixed with the feed which are supposed to bind the mycotoxins efficiently in the gastro-intestinal tract. Aluminosilicates are the preferred adsorbents, followed by activated charcoal and special polymers. The efficiency of mycotoxin binders, however, differs considerably depending mainly on the chemical structure of both the adsorbent and the toxin. This review describes the most important types of adsorbents and the respective mechanisms of adsorption. Data of the in vitro and in vivo efficacy of detoxication are given.

Adsorption↗