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PubMed2002

Update of survey, regulation and toxic effects of mycotoxins in Europe.

The most frequent toxigenic fungi in Europe are Aspergillus, Penicillium and Fusarium species. They produce aflatoxin B1 transformed into aflatoxin M1 found in the milk, as well as Ochratoxins and Zearalenone, Fumonisin B1, T-2 toxin, HT-2 toxin and deoxynivalenol (vomitoxin), which are of increasing concern in human health. These mycotoxins are under continuous survey in Europe, but the regulatory aspects still need to be set up and/or harmonised at European level. They are found in foodstuffs and are not destroyed by normal industrial processing or cooking since they are heat-stable. Some of their metabolites are still toxic and may be involved in human diseases. Their toxic effects (liver, kidney and hematopoetic toxicity, immune toxicity, reproduction toxicity, foetal toxicity and teratogenicity, and mainly carcinogenicity) are mostly known in experimental models, the extrapolation to humans being always inaccurate. The inaccuracy of extrapolation to humans may be explained by the lack of adequate food consumption data, lack of knowledge about relative health risks associated with specifically proposed limits and by the possibility of synergism with other mycotoxins present in the same food commodities. Other pathological causes are viral hepatitis, immune or hormonal deficiencies or organ dysfunction. Even when a specific biomarker of a given mycotoxin is identified in humans, it remains difficult to establish the relation with a given illness, because of genetic polymorphism and the possible beneficial influence of diet, and because other environmental toxicants may well interfere. The acceptable daily intake limits are mostly based on animal data and may be too high, due to the differences in the sensitivity of different animal species. The prevention involves first reduction of mycotoxin levels in foodstuffs and further increasing the intake of diet components such as vitamins, antioxidants and substances known to prevent carcinogenesis.

Aflatoxins↗

PubMed2001

Effects of mycotoxins on human immune functions in vitro.

Immunosuppressive and carcinogenic Fusarium mycotoxins may appear in domestic food products. Therefore, the immunological effects of Fusarium mycotoxins were tested on human peripheral blood mononuclear cells from different blood donors. In the present study we investigated deoxynivalenol (DON), 3-acetyldeoxynivalenol, fusarenon-X, T-2 toxin, zearalenone, alpha-zearalenol, beta-zearalenol and nivalenol for their effects on T and B cells in a proliferation assay, antibody-dependent cellular cytotoxicity (ADCC) and natural killer (NK) cell activity on human peripheral blood mononuclear cells. The concentrations applied in our experiments were similar to those which can be found in normal human peripheral blood system (0.2--1800 ng/ml). Among the eight mycotoxins tested, T-2 toxin, fusarenon X, nivalenol and deoxynivalenol exerted the highest immunosuppressing effect on human peripheral blood mononuclear cells in vitro. Mycotoxin-induced immunosupression was manifested as depressed T or B lymphocyte activity. Furthermore, by virtue of inhibition of NK cell activity, the protection against tumor development may also be attenuated.

Antibody Formation↗

PubMed1999

Increased cytotoxicity of food-borne mycotoxins toward human cell lines in vitro via enhanced cytochrome p450 expression using the MTT bioassay.

Eight food-borne mycotoxins epidemiologically implicated in human disease were tested for their cytotoxic effects on human cells previously immortalised and transfected to introduce human cytochrome p450 (CYP 450) genes. Such cells retain many characteristics of normal cell growth and differentiation while simultaneously having the potential of either increasing or decreasing the metabolic activity (cytotoxicity) of the challenging mycotoxins. The MTT assay provided an indication of cytotoxicity. Of the nine CYP450s introduced CYP1A2 was most effective, rendering the cells 540 times more sensitive than the control cells to aflatoxin B1, 28 times more sensitive to aflatoxin G1 and 8-fold more sensitive to ochratoxin A. CYP3A4 resulted in the cells being 211 times more toxic to aflatoxin B1 and 8-fold more toxic to aflatoxin G1 while CYP 2A6, CYP 3A5 and CYP 2E1 also produced observable effects. No increase in metabolic activity was found using cyclopiazonic acid, deoxynivalenol, fumonisin B1, patulin or T-2 toxin. CD5Os were calculated for the mycotoxins against the non-CYP-introduced control cells. There was almost a five order of magnitude difference between the most toxic, T-2 toxin (CD50 0.0057 microgram/ml) and the least toxic, fumonisin B1 (CD50 476.2 micrograms/ml). In vitro biological assays thus provide an excellent system for quantifying the often low CD50s expressed by mycotoxins in foods.

Biological Assay↗

PubMed1998

Moulds and mycotoxins in herb tea and medicinal plants.

The level of toxigenic moulds and mycotoxins were analyzed in 62 samples of medicinal plant material and 11 herbal tea samples. The most predominant fungi detected were: Aspergillus, Penicillium, Mucor, Rhizopus, Absidia, Alternaria, Cladosporium and Trichoderma. Aspergillus flavus, a known producer of the aflatoxin mycotoxin, was present in 11 or 18% of the 62 medicinal plant samples and in 1 or 9% of the herbal tea samples. The medicinal plant samples, contaminated with A. flavus were also analyzed for the mycotoxins aflatoxin, ochratoxin and zearalenone; ochratoxin was found in one of the 7 samples analyzed. This study suggests that medicinal plant material and possibly herbal teas, if stored improperly allowing for mould growth, should be analyzed for mould and mycotoxin prior to use.

Aflatoxins↗

PubMed2000

The effect of combinations of Fusarium mycotoxins (deoxynivalenol, zearalenone and fumonisin B1) on growth of brewing yeasts.

The interactive effect of combinations of the Fusarium mycotoxins deoxynivalenol (DON), zearalenone (ZEA) and fumonisin B1 (FB1) on growth of brewing yeasts was examined. Yeast growth was assessed by measurement of dry weight or relative growth, cell number, viability and conductance change of the growth medium using direct and indirect methods. The interactive effect of a combination of these mycotoxins was subject to the ratio of toxins in the mixture and the toxicity of individual toxins on yeast growth. When a combination of mycotoxins at low concentration was added into the growth medium, no significant inhibitory effect on growth was observed compared to controls. However, when a combination of high concentrations of DON and ZEA which individually inhibited yeast growth was examined, the interactive effect was shown to pass from antagonism to synergism depending on the ratio of the toxins in the mixture. As a synergistic interaction between these Fusarium mycotoxins was observed only at high concentrations, which were far higher than would be expected in good quality grain, they are not a concern when related to yeast growth under the brewing conditions studied.

Beer↗

PubMed2001

Mycotoxin production from fungi isolated from grapes.

AIMS: In order to assess the potential for producing mycotoxins, fungi were isolated from wine producing grapes. METHODS AND RESULTS: The isolates were identified and Penicillium expansum, the most well recognized mycotoxin producer, was analysed for mycotoxin production by TLC. Many of the strains produced patulin and/or citrinin, often depending on whether they were grown on a grape or yeast extract sucrose media. CONCLUSION: Citrinin was produced by all strains grown in the yeast extract sucrose medium, but only one strain (from 51) was able to produce this compound in grape juice medium. Patulin was produced in the yeast extract medium by 20 strains and in grape juice medium by 33 strains. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of mycotoxins in wine producing grapes is discussed. Grapes contamination with patulin seems not to contribute to wine contamination, and no ochratoxin producing fungi was identified.

Beverages↗

PubMed1996

Fumonisins and Alternaria alternata lycopersici toxins: sphinganine analog mycotoxins induce apoptosis in monkey kidney cells.

Fusarium moniliforme toxins (fumonisins) and Alternaria alternata lycopersici (AAL) toxins are members of a new class of sphinganine analog mycotoxins that occur widely in the food chain. These mycotoxins represent a serious threat to human and animal health, inducing both cell death and neoplastic events in mammals. The mechanisms by which this family of chemical congeners induce changes in cell homeostasis were investigated in African green monkey kidney cells (CV-1) by assessing the appearance of apoptosis, cell cycle regulation, and putative components of signal transduction pathways involved in apoptosis. Structurally, these mycotoxins resemble the sphingoid bases, sphingosine and sphinganine, that are reported to play critical roles in cell communication and signal transduction. The addition of fumonisin B1 or AAL toxin, TA, to CV-1 cells induced the stereotypical hallmarks of apoptosis, including the formation of DNA ladders, compaction of nuclear DNA, and the subsequent appearance of apoptotic bodies. Neither mycotoxin induced cell death, DNA ladders, or apoptotic bodies in CV-1 cells expressing simian virus 40 large T antigen (COS-7) at toxin concentrations that readily killed CV-1 cells. Fumonisin B1 induced cell cycle arrest in the G1 phase in CV-1 cells but not in COS-7 cells. AAL toxin TA did not arrest cell cycle progression in either cell line. The induction of apoptosis combined with the widespread presence of these compounds in food crops and animal feed identifies a previously unrecognized health risk to humans and livestock. These molecules also represent a new class of natural toxicants that can be used as model compounds to further characterize the molecular and biochemical pathways leading to apoptosis.

Alternaria↗

PubMed1999

Mycotoxins: their implications for human and animal health.

Mycotoxins contaminate various feed and food commodities, due to the global occurrence of toxinogenic molds. They exert adverse health effects in human and animals. The nature of these toxic effects varies depending on the chemical structure of the toxin. The degree of these adverse effects is not only determined by the toxin concentration present in foods and feeds, but also by the time of exposure. Whilst in animals, next to acute intoxication, losses in productivity, reduced weight gain and immunosuppression are considered as most important feature of mycotoxicoses, genotoxic effects and the involvement of certain mycotoxins such as aflatoxin, ochratoxins and fumonisins in the etiology of human cancers have obtained particular attention. This implies that recent research activities concentrate on mechanistic aspects of mycotoxin-induced pathologies, rather than compiling analytical measures of mycotoxin concentrations in food and feeds.

Animal Feed↗

PubMed2004

Presence and concentrations of the Fusarium-related mycotoxins beauvericin, enniatins and moniliformin in finnish grain samples.

Fusarium mycotoxins beauvericin, enniatins (A, A1, B, B1) and moniliformin were analysed in 38 Finnish grain samples (14 wheat, 22 barley, one rye, one oats) harvested in 2001-02. The contaminating Fusarium species were identified with the primer-specific polymerase chain reaction as well as with morphological studies. All the studied mycotoxins were found in the samples. Enniatins B and B1 were detected in all samples, and enniatin A, enniatin A1, beauvericin and moniliformin in 74, 95, 95 and 74% of the samples, respectively. There were higher concentrations of the mycotoxins analysed in 2001 compared with 2002. The highest levels of mycotoxins were detected in samples harvested late in the autumn after a long rainy period. Fusarium avenaceum was the most abundant Fusarium species in Finland during both years (0-29.5%) measured as infected kernels. A significant correlation was found between F. avenaceum contamination level and the concentration levels of enniatins B and B1, as well as moniliformin.

Cyclobutanes↗

PubMed2005

Fate of mycotoxins in cereals during extrusion cooking: a review.

Extrusion cooking is one of the fastest growing food-processing operations in recent years due to several advantages over traditional methods. Apart from its main goal of improving the quality of intermediate and final processed products, it may incidentally also improve safety because of the potential to reduce mycotoxin levels in cereals. This review is focused on extrusion cooking and aims to give a general overview of its impact in reducing mycotoxin levels in cereals. Extrusion cooking generally decreases the mycotoxins levels at rates depending on different factors such as the type of extruder, the type of screw, the die configuration, the initial mycotoxin concentration, the barrel temperature, the screw speed, the moisture content of the raw material and the use of additives. Reductions of 100, 95 and 83% for fumonisins, aflatoxins and zearalenone, respectively, have been reported during extrusion cooking of cereals, while lower reductions were observed for deoxynivalenol, ochratoxin A and moniliformin, where maximum reductions did not exceed 55, 40 and 30%, respectively.

Aflatoxins↗

PubMed2005

Determination of selected mycotoxins in mould cheeses with liquid chromatography coupled to tandem with mass spectrometry.

A simple and feasible method is described for analysing nine mycotoxins in cheese matrix. The method involves liquid extraction followed by high performance liquid chromatographic separation and mass spectrometric detection of the analytes, and allows the determination of aflatoxins B1, B2, G1, G2 and M1, ochratoxin A, mycophenolic acid, penicillic acid and roquefortine C simultaneously. Average recoveries of the mycotoxins from spiked samples at concentration levels of 5-200 microg kg(-1) ranged from 96-143%. Within-day relative standard deviations at these concentration levels varied from 2.3-12.1%. The limit of quantification for aflatoxin M1 was 0.6 microg kg(-1) and for the other compounds 5 microg kg(-1). The method developed was applied for analysing these mycotoxins in blue and white mould cheeses purchased from Finnish supermarkets. Roquefortine C was detected in all of the blue mould cheese samples in concentrations of 0.8-12 mg kg(-1). One blue cheese contained also 0.3 mg kg(-1) mycophenolic acid. The other investigated mycotoxins were absent in the samples.

Aflatoxin B1↗

PubMedDate not supplied

Rationales for the establishment of limits and regulations for mycotoxins.

Although 50 countries have enacted or proposed regulations for control of alfatoxins in food or feed, and 15 of these countries also have regulations for permitted levels of contamination by other mycotoxins, very few countries have formally presented the rationale for the need to regulate, or for the selection of a particular maximum tolerated level. After several successive inquiries, information concerning the rationale for regulation was obtained from 21 countries. Most of the responses concerned limits for aflatoxin in food, and most of these were based on a vague, unsupported statement of the carcinogenic risk for humans. There was a general consensus that exposure to a potential human carcinogen that could not be totally avoided should be limited to the lowest practical level; the definition of practicality depended on whether the country was an importer or producer of the potentially contaminated commodity. A claim to a hazard evaluation was made by six countries (Canada, Belgium, India, United Kingdom, United States, Switzerland) without providing specifics; and one country, South Africa, referred to a risk determination. The most comprehensive rationale for any mycotoxin regulation was provided by the United States in support of limits for aflatoxin in specific animal feedstuffs. The responses provided no rationale for setting limits for other mycotoxins; but scholarly risk assessments for zearalenone and ochratoxin A have been published by Canadian government scientists, and a symposium presentation provides the information that in Norway patulin is regulated for quality control purposes only. It is apparent that, in most countries, either the scientific basis for regulation of mycotoxins is nonexistent, or the science has not been fully utilized.(ABSTRACT TRUNCATED AT 250 WORDS)

Aflatoxins↗

PubMedDate not supplied

Mycotoxins in review.

The putative involvement of mycotoxins in human diseases, including cancer, is reviewed with reference to ergotism, citreoviridin toxicosis, alimentary toxic aleukia, stachybotryotoxicosis, Balkan endemic nephropathy and aflatoxicoses. Analytical and sampling problems in determining the occurrence of and exposure to mycotoxins are discussed against the background that over 300 mycotoxins have been identified, produced by some 350 fungal species, and that the potency of some of them demands the quantitation of extremely small quantities of analyte. Mycotoxins associated with food and originating from Alternaria, Aspergillus, Fusaria, Penicillia and Claviceps spp. are reviewed with reference to the toxicological, regulatory and economic issues arising. It is concluded that continued efforts are needed to: identify and quantify human/animal exposure; estimate health risks and make defensible risk-benefit judgements; develop sampling plans based on experimental observation; arrive at agreed regulatory levels based on legitimate sampling plans, analytical capabilities and economic considerations; develop procedures for disposal of contaminated lots; and develop plant varieties resistant to fungal invasion.

Food Contamination↗

PubMedDate not supplied

Occurrence of mycotoxins in raw ingredients used for animal feeding stuffs in the United Kingdom in 1992.

Examination of 330 samples of animal feed ingredients for the presence of a number of mycotoxins has been carried out. These samples were drawn from 186 animal feed mills in the United Kingdom. Aflatoxin B1 was the mycotoxin found most frequently, occurring in most samples of rice bran, maize products, palm kernels and cottonseed, but not in only 3 out of 20 samples of sunflower, in 1 out of 20 samples of soya and in no samples of peas, beans or manioc. Analytical difficulties were met with some combinations of commodity and mycotoxin and all results are uncorrected for recovery. The highest level was detected in a sample of maize gluten: 41 micrograms/kg of aflatoxin B1 (47 micrograms/kg total aflatoxins). Maize products also frequently contained fumonisins B1 and B2 at levels up to nearly 5,000 micrograms/kg in total and zearalenone up to a maximum level of 500 micrograms/kg. Ochratoxin A and citrinin were found in approximately 20% of wheat and barley samples. One sample of barley contained ochratoxin A at a level of 102 micrograms/kg and citrinin at a level of 8 micrograms/kg. Low levels of ochratoxin A also occurred in a few samples of other ingredients: rice bran, palm kernel and beans. Sterigmatocystin at 18 micrograms/kg was found in one sample of organically grown wheat and a trace amount of zearalenone in one sample of manioc. Multi-mycotoxin contamination also occurred, particularly in some samples of maize for which 19 out of 50 samples contained both aflatoxins and fumonisins.

Animal Feed↗

PubMed2006

Effects of feeding grains naturally contaminated with Fusarium mycotoxins on brain regional neurochemistry of laying hens, turkey poults, and broiler breeder hens.

Three experiments were conducted to compare the effects of feeding blends of grains naturally contaminated with Fusarium mycotoxins on brain regional neurochemistry of laying hens, turkey poults, and broiler breeder hens. In Experiment 1, thirty-six 45-wk-old laying hens were fed diets including the following for 4 wk: 1) control, 2) contaminated grains, and 3) contaminated grains + 0.2% polymeric glucomannan mycotoxin adsorbent (GMA). Concentrations of brain neurotransmitters and metabolites were analyzed in pons, hypothalamus, and cortex by HPLC with electrochemical detection. Neurotransmitters and the metabolites measured included dopamine, 3,4-dihydroxylphenyacetic acid, homovanillic acid, serotonin [5-hydroxytryptamine (5-HT)], 5-hydroxyindolacetic acid, epinephrine, and norepinephrine. The feeding of contaminated grains significantly increased concentrations of 5-HT and decreased the 5-hydroxyindolacetic acid:5-HT in the pons region in the brain stem. Dietary supplementation with GMA prevented these effects. There was no effect of diet on concentrations of other neurotransmitters or metabolites in the pons, hypothalamus, or cortex. In Experiment 2, thirty-six 1-d-old turkey poults were fed diets including the following for 4 wk: 1) control, 2) contaminated grains, and 3) contaminated grains + 0.2% GMA. Hypothalamic, pons, and cortex neurotransmitter concentrations were not affected by diet. In Experiment 3, forty-two 26-wk-old broiler breeder hens were fed diets including the following for 15 wk: 1) control, 2) contaminated grains, and 3) contaminated grains + 0.2% GMA. There was no effect of diet on neurotransmitter concentrations in the pons, hypothalamus, or cortex. It was concluded that differences in intraspecies effects of these mycotoxins on brain neurotransmitter concentrations might explain the intraspecies differences in the severity of Fusarium mycotoxin-induced reductions in feed intake.

Animal Feed↗

PubMed2003

Clinical implications of mycotoxins and Stachybotrys.

There has been considerable interest and concern in recent years regarding the potential health effects of mycotoxins in the indoor environment. Although the existence of mycotoxins has been known for several decades, relatively little is known about their effects in humans. What is known comes almost exclusively from studies of ingestion as the route of exposure. This review summarizes what is known regarding health effects of mycotoxins in general and specifically examines the evidence for the role of indoor exposure to the fungi of the genus Stachybotrys as a cause of disease in humans. Much work remains to be done in the area of mycotoxin research. The risk of health effects from ingestion seems much more widespread than from indoor airborne exposure, although the latter has received considerably more media attention. Rigorously controlled studies are needed to clarify these issues.

Aflatoxins↗

PubMed2004

Mycotoxins and antifungal drug interactions: implications in the treatment of illnesses due to indoor chronic toxigenic mold exposures.

Chronic exposure to toxigenic molds in water-damaged buildings is an indoor environmental health problem to which escalating health and property insurance costs are raising a statewide concern in recent times. This paper reviews the structural and functional properties of mycotoxins produced by toxigenic molds and their interactive health implications with antifungal drugs. Fundamental bases of pathophysiological, neurodevelopmental, and cellular mechanisms of mycotoxic effects are evaluated. It is most likely that the interactions of mycotoxins with antifungal drugs may, at least in part, contribute to the observable persistent illnesses, antifungal drug resistance, and allergic reactions in patients exposed to chronic toxigenic molds. Safe dose level of mycotoxin in humans is not clear. Hence, the safety regulations in place at the moment remain inconclusive, precautionary, and arbitrary. Since some of the antifungal drugs are derived from molds, and since they have structural and functional groups similar to those of mycotoxins, the knowledge of their interactions are important in enhancing preventive measures.

Air Pollution, Indoor↗

PubMed2000

Fusarium species from nepalese rice and production of mycotoxins and gibberellic acid by selected species.

Infection of cereal grains with Fusarium species can cause contamination with mycotoxins that affect human and animal health. To determine the potential for mycotoxin contamination, we isolated Fusarium species from samples of rice seeds that were collected in 1997 on farms in the foothills of the Nepal Himalaya. The predominant Fusarium species in surface-disinfested seeds with husks were species of the Gibberella fujikuroi complex, including G. fujikuroi mating population A (anamorph, Fusarium verticillioides), G. fujikuroi mating population C (anamorph, Fusarium fujikuroi), and G. fujikuroi mating population D (anamorph, Fusarium proliferatum). The widespread occurrence of mating population D suggests that its role in the complex symptoms of bakanae disease of rice may be significant. Other common species were Gibberella zeae (anamorph, Fusarium graminearum) and Fusarium semitectum, with Fusarium acuminatum, Fusarium anguioides, Fusarium avenaceum, Fusarium chlamydosporum, Fusarium equiseti, and Fusarium oxysporum occasionally present. Strains of mating population C produced beauvericin, moniliformin, and gibberellic acid, but little or no fumonisin, whereas strains of mating population D produced beauvericin, fumonisin, and, usually, moniliformin, but no gibberellic acid. Some strains of G. zeae produced the 8-ketotrichothecene nivalenol, whereas others produced deoxynivalenol. Despite the occurrence of fumonisin-producing strains of mating population D, and of 8-ketotrichothecene-producing strains of G. zeae, Nepalese rice showed no detectable contamination with these mycotoxins. Effective traditional practices for grain drying and storage may prevent contamination of Nepalese rice with Fusarium mycotoxins.

Carboxylic Acids↗