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Maize flour contaminated with toxigenic fungi and mycotoxins in Kenya.

The majority of the Kenyan human population may be exposed to regular doses of a wide spectrum of highly toxic, carcinogenic, immunosuppressive, mutagenic, and hepatotoxic mycotoxins through the consumption of maizemeal. Maizemeal is consumed in Kenya at the rate of about 0.4kg/person/day, therefore, even the lowest amount of toxins consumed can cause significant effects as some are cumulative. Maize is also a major component in livestock and poultry feed, and the therefore regular indirect human exposure through the consumption of animal products that contain mycotoxin residues could be common. Due to these suspected risks, it is imperative to establish whether consumers in Kenya are actually exposed to dangerous mycotoxins in maize products hence this survey. Forty samples of flour packed in 90kg bags, 58 samples of "Ugali" brand (milled and packed by the Milling Corporation of Kenya Ltd), and 74 samples of "Jogoo" brand ( Unga Maize Millers) were collected from the Nairobi area. The samples were analysed for resident mycoflora, and some mycotoxins associated with key fungal species. Important fungal species isolated from the flour included Aspergillus flavus, A. sulphureus, Fusarium moniliforme, Penicillium stoloniferum, and P. cyclopium. All the three brands of flour were contaminated with Aflatoxins B1 and B2 (0.4-20 ug/kg), Ochratoxin A(50-1,500 ug/kg), and Zearalenone (2,500 - 5,000 ug/kg). Ochratoxin A was the most prevalent mycotoxin. These data provide a warning that the mycotoxin contamination problem in maizemeal is critical and consumers' health is at risk. Therefore, rigorous countrywide monitoring of mycotoxins in this staple food should be pursued. If possible, maize products should be subjected to stiff microbial quality control from the farm gate to the market shelf.

Journal Article↗

[The diagnosis of mycotoxin hepatitis in swine].

Mycotoxin hepatitis which develops in pigs when they eat fodder contaminated by mycotoxin (sterigmatocystin) is registered at large pig breeding farms of Ukraine. Diagnostics of mycotoxin hepatitis of pigs induced by sterigmatocystin is based on clinical-epizootiological studies and mycotoxin analysis of the fodder including detection of fungi, producers of sterigmatocystin and determination of the content of mycotoxin itself. The authors have suggested a method based on extraction of mycotoxin from the fodder by means of the mixture of chloroform and 4% water solution of potassium chloride, on concentration of the chloroform residue, its separation in a thin layer of silicagel, and UV examination of chromatograms. The method sensitivity is 30 micrograms per 1 kg of the product. This method will permit diagnosis of mycotoxin hepatitis in animals and will promote timely prophylactic measures. The method is easy to be applied in veterinary laboratories.

Animal Feed↗

Monitoring program for mycotoxin contamination in Uruguayan food and feeds.

A pilot study for monitoring mycotoxin contamination in food and feeds was implemented by the Technological Laboratory of Uruguay (LATU) with the technical assistance of the Food and Agriculture Organization of the United Nations (FAO). The scope of the study was to determine the potential hazard posed by priority food-contaminant and feed-contaminant combinations. The choice of foods and contaminants to be monitored was based on the importance of the food in the total diet, the economic importance of the product and the potential health risk posed by the specific combination. The principal commodities selected were wheat, barley and rice. Also included were com, soy, dairy products, feeds, dried fruits and legumes, oil seeds, cocoa beans and organ meats. Mycotoxins analyzed (TLC/densitometry) were aflatoxins, zearalenone, ochratoxin A, deoxynivalenol (DON) and ergot alkaloids. The survey results (1993-95) showed differences in both incidence and levels of mycotoxin content for the principal commodities. Of all food/feed categories analyzed, feed had the highest values for all mycotoxins. Samples containing DON in levels above 1000 ng/g were found in all groups. Ochratoxin A was not detected in any of the samples. Rice and soy beans were the categories with lowest aflatoxin incidence. Uruguayan regulatory limits for all toxins analyzed were exceeded for wheat, barley and rice in less than 3, 9 and 7% of samples, respectively. The data on actual mycotoxin levels in different foods will help identify sources of contaminations and areas where control measures should be improved, enable better risk assessment by proper estimation of mycotoxin intake, assist in the establishment of tolerances and adequate guidelines, aid in the implementation of a national program and provide economic benefits by improving grain quality.

Aflatoxins↗

Chemiluminescent and bioluminescent assays as innovative prospects for mycotoxin determination in food and feed.

Mycotoxin contamination of food and feedstuffs is among the top priorities for human and animal safety. The currently used techniques for mycotoxin determination, either chromatography or ELISA, are unsuitable for routine in-field assessment. There is an urgent need for other accurate, simple and cost-effective techniques that can be used as a screening tool for a rapid estimation of mycotoxin contamination in commodity lots. This paper reviews the literature on the use of chemiluminescence (CL) and bioluminescence (BL) assays for direct or indirect mycotoxin assessment. The chemiluminescence immunoassays, adenosine triphosphate (ATP) bioluminescence and bioassays are reviewed and their advantages and limitations discussed. These techniques used in food testing and the pharmaceutical industry offer promise as rapid techniques for mycotoxin determination. Chemiluminescence and bioluminescence bioassays are the most innovative alternatives to the conventional techniques used for mycotoxin determination in food and feed.

Adenosine Triphosphate↗

Monitoring the mycotoxins in food and their biomarkers in the Czech Republic.

Testing of the presence of toxigenic microfungi and mycotoxins in foodstuffs in the food chain is an important part of the food safety strategy in The Czech Republic. At the national level, control of their presence in the entire food chain is assured by Public Health Protection Agencies, by the Veterinary Administration and by the Czech Agriculture and Food Inspection Authority. This article summarizes surveillance activities of Public Health Protection Agencies and mycotoxins findings in dietary raw materials and foodstuffs from the 1990s to 2004 in the Czech Republic. At present, the health risk from the mycotoxins exposure from foodstuffs is assessed to be relatively low in the Czech Republic, especially as far as the foodstuffs of the Czech origin are concerned. It may result in late toxic effects (e. g., carcinogenic risk) following a single or repeated ingestion of low mycotoxins doses from foodstuffs. Nevertheless, the overall situation may change due to the globalization of the food market. In order to minimize the risk associated with mycotoxins and eliminate their impact on Czech public health, continuous monitoring of the presence of toxigenic moulds, mycotoxins, and their biomarkers is necessary, in conjunction with strict respect to European Union legislation.

Aflatoxin B1↗

FAO programmes for prevention, regulation, and control of mycotoxins in food.

The article provides information on recent past and current activities carried out by the Food and Agriculture Organization of the United Nations (FAO) at national, regional, and international levels to assist member countries in overcoming the problems raised by mycotoxins in foods and feeds. It gives special emphasis to the preventive aspects at the stages of production and storage of those commodities which are particularly sensitive to mycotoxin contamination. Specific projects implemented by the FAO in various countries to prevent and control mycotoxin contamination are cited with particular reference to the on-going FAO/United Nations Environment Programme (UNEP) Regional Training Network for Mycotoxin Control in Asia. The article also provides an update of the Codex Alimentarius effort to establish international guideline levels for different mycotoxins in various foodstuffs and agricultural commodities and to develop sampling plans for aflatoxin control in peanuts and maize and their products. It proposes a set of recommended actions at various levels to monitor and control mycotoxin levels in foods and to promote international cooperation in this field.

Animals↗

A practical guide to the prevention of Fusarium mycotoxins in grain and animal feedstuffs.

The guidelines in this report focus on prevention of mycotoxin contamination associated with plant and animal production in eastern Canada. The topics covered include a brief description of the Fusarium mycotoxins of particular concern in eastern Canada, the life cycle of Fusarium and recommendations for reducing fungal infection and mycotoxin production in grain crops and animal feed. The presence of complex mixtures of toxins, as well as research on detoxification and mycotoxin residues in animal food products are discussed. For those involved in advising producers and millers on occupational hazards associated with mycotoxins, guidelines on the safe handling of contaminated grain are also included. The review provides background information for scientists and technical personnel advising grain and animal producers on how to reduce mycotoxin problems associated with F. graminearum contamination of grain, especially wheat and corn. It may be necessary to revise the recommendations to suit different agricultural regions or different plant and animal production systems.

Animal Feed↗

Trace mycotoxin analysis in complex biological and food matrices by liquid chromatography-atmospheric pressure ionisation mass spectrometry.

Mycotoxins are toxic secondary metabolites produced by filamentous fungi that are growing on agricultural commodities. Their frequent presence in food and their severe toxic, carcinogenic and estrogenic properties have been recognised as potential threat to human health. A reliable risk assessment of mycotoxin contamination for humans and animals relies basically on their unambiguous identification and accurate quantification in food and feedstuff. While most screening methods for mycotoxins are based on immunoassays, unambiguous analyte confirmation can be easily achieved with mass spectrometric methods, like gas chromatography/mass spectrometry (GC/MS) or liquid chromatography/mass spectrometry (LC/MS). Due to the introduction of atmospheric pressure ionisation (API) techniques in the late 80s, LC/MS has become a routine technique also in food analysis, overcoming the traditional drawbacks of GC/MS regarding volatility and thermal stability. During the last few years, this technical and instrumental progress had also an increasing impact on the expanding field of mycotoxin analysis. The aim of the present review is to give an overview on the application of LC-(API)MS in the analysis of frequently occurring and highly toxic mycotoxins, such as trichothecenes, ochratoxins, zearalenone, fumonisins, aflatoxins, enniatins, moniliformin and several other mycotoxins. This includes also the investigation of some of their metabolites and degradation products. Suitable sample pre-treatment procedures, their applicability for high sample through-put and their influence on matrix effects will be discussed. The review covers literature published until July 2006.

Aflatoxins↗

Effects of the mycotoxins alpha- and beta-zearalenol on regulation of progesterone synthesis in cultured granulosa cells from porcine ovaries.

Mycotoxins as contaminants of animal food can impair fertility and can cause abnormal fetal development in farm animals. Therefore, the present study has investigated whether derivatives of the mycotoxin zearalenone, alpha-zearalenol (alpha-ZOL) and beta-zearalenol (beta-ZOL), influence progesterone synthesis via cytochrome p450 side chain cleavage enzyme (p450scc) and 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD) in cultured porcine granulosa cells. Both enzymes are essential for the conversion of cholesterol to progesterone. No differences in basal progesterone levels and numbers of viable cell were observed between untreated granulosa cells and those treated with alpha- or beta-ZOL (15 and 30 microM). FSH (0.01 microg/ml) or forskolin (10 microM) enhanced the basal progesterone secretion in the absence of mycotoxins. The addition of alpha- or beta-ZOL (7.5, 15 and 30 microM) to cultures stimulated with FSH (0.01 microg) or forskolin (10 microM) reduced progesterone synthesis and the levels of p450scc and 3beta-HSD transcripts in a dose-dependent manner (P<0.05). The enzymatic activity of 3beta-HSD and the abundance of p450scc protein were also reduced by these mycotoxins. In conclusion, effects of mycotoxins on FSH receptor-dependent and receptor-independent pathways indicate that adenylate cyclase activity and/or regulatory pathways further downstream are targets of mycotoxin actions. The apparent dose-dependent reduction of p450scc and 3beta-HSD transcripts implies an effect of alpha- and beta-ZOL on transcriptional regulation of these enzymes.

3-Hydroxysteroid Dehydrogenases↗

Antioxidant nutrients and mycotoxins.

Mycotoxins are fungal secondary metabolites that are toxic to vertebrates produced by organisms that occur as plant pathogens, soilborne fungi, airborne fungi and aeroallergens. They are distributed worldwide and may be recovered from a wide range of substrates. Their presence in food and feeds, as the result of fungal diseases in crops, can present a danger to animal and human health. Many mycotoxins have also been shown to be phytotoxic and in some cases, such as with trichothecenes produced by the wheat head blight fungus Fusarium graminearum, mycotoxins may act as virulence factors. Several natural (vitamin, provitamins, carotenoids, chlorophyll and its derivatives, phenolics, and selenium) and synthetic (butylated hydroxyanisole and butylated hydroxytoluene) compounds with antioxidant properties seem to be potentially very efficacious in protecting against the toxic effects of mycotoxins. The protective properties of antioxidants are probably due to their ability to act as superoxide anion scavengers, thereby protecting cell membranes from mycotoxin-induced damage and in some cases, antioxidant vitamins may play a role in preventing mycotoxicosis. However, much less information is available from studies carried out on antioxidants and mycotoxins, such as OTA, FB(1), T-2 toxin, ZEN, and citrinin. No such studies have been performed on recently discovered toxins such as beauvericin, fusaproliferin, moniliformin, and fusaric acid. However, supplementation with antioxidant nutrients to prevent mycotoxicosis has been controversial. The case for the use of supplemental antioxidant vitamins at the present time needs further research.

Animals↗

Mycotoxins. Contemporary issues of food animal health and productivity.

Modern agriculture and production animal medicine require attention to high-quality feeds that are free from mycotoxin contamination that can cause economically important decreases in productivity. Maintaining current information about effects of mycotoxins on feed intake and growth, reproductive efficiency, and possible immunosuppression aid in effective consultation with livestock producers. Investigation and determination of potential production losses related to mycotoxins should use historical, clinical, laboratory, and experimental information to objectively evaluate whether mycotoxin contamination is clinically relevant. The practicing veterinarian or veterinary consultant can provide valuable clinical and interpretive assistance to producers who may have a real or potential mycotoxin contamination. Thorough diagnostic evaluation of animals, appropriate testing of feeds and forages, and rational consideration of differential diagnoses help to put mycotoxins in the proper perspective as a production-related management problem.

Animal Feed↗

Current situation on regulations for mycotoxins. Overview of tolerances and status of standard methods of sampling and analysis.

A worldwide enquiry was undertaken in 1986-1987 to obtain up-to-date information about mycotoxin legislation in as many countries of the world as possible. Together with some additional data collected in 1981, information is now available about planned, proposed, existing or absence of legislation in 66 countries. Details about tolerances, legal bases, responsible authorities, prescribed methods of sampling and analysis and disposition of commodities containing inadmissible amounts of mycotoxins, are given. The information concerns aflatoxins in foodstuffs, aflatoxin M1 in dairy products, aflatoxins in animal feedstuffs, and other mycotoxins in food- and feedstuffs. In comparison with the situation in 1981, limits and regulations for mycotoxins have been expanded in 1987 with more countries having legislation (proposed or passed) on the subject, more products, and more mycotoxins covered by this legislation. The differences between tolerances in the various countries are sometimes quite large, which makes harmonization of mycotoxin regulations highly desirable.

Global Health↗

Risk from inhaled mycotoxins in indoor office and residential environments.

Mycotoxins are known to produce veterinary and human diseases when consumed with contaminated foods. Mycotoxins have also been proposed to cause adverse human health effects after inhalation exposure to mold in indoor residential, school, and office environments. Epidemiologic evidence has been inadequate to establish a causal relationship between indoor mold and nonallergic, toxigenic health effects. In this article, the authors model a maximum possible dose of mycotoxins that could be inhaled in 24 h of continuous exposure to a high concentration of mold spores containing the maximum reported concentration of aflatoxins B1 and B2, satratoxins G and H, fumitremorgens B and C, verruculogen, and trichoverrols A and B. These calculated doses are compared to effects data for the same mycotoxins. None of the maximum doses modeled were sufficiently high to cause any adverse effect. The model illustrates the inefficiency of delivery of mycotoxins via inhalation of mold spores, and suggests that the lack of association between mold exposure and mycotoxicoses in indoor environments is due to a requirement for extremely high airborne spore levels and extended periods of exposure to elicit a response. This model is further evidence that human mycotoxicoses are implausible following inhalation exposure to mycotoxins in mold-contaminated home, school, or office environments.

Adult↗

Aspergillus mycotoxins and their effect on the host.

Aspergillus fumigatus is known to produce various immunosuppressive mycotoxins including gliotoxin. However, none of these mycotoxins has been confirmed as being directly related to the pathogenesis of aspergilli. Recent studies have made substantial progress in the determination of mycotoxins as virulence factors. Gliotoxin was found to be produced much faster than previously believed under certain culture conditions, such as at 37 degrees C and under high oxygen content, which is close to the environment in the host. Gliotoxin was also found to be detectable in the sera of aspergillosis mice and of aspergillosis patients. Based on these findings, it is becoming evident that gliotoxin is produced in the infected organs of patients of aspergillosis at a significant level. In addition to these known mycotoxins, A. fumigatus produces many mycotoxins apparently different from known toxins. From the aspect of gene analysis, the deletion of laeA was found to block the expression of metabolic gene clusters such as sterigmatocystin, and the gene is also expected to be related to the production of gliotoxin. The significance of mycotoxins as virulence factors will hopefully be clarified in the near future.

Animals↗

The influence of contamination with separate mycotoxins (aflatoxins, ochratoxin A, citrinin, patulin, penicillic acid or sterigmatocystin) on the in vitro dry matter and organic matter digestibilities of some roughages (berseem hay and wheat straw).

In vitro study on berseem hay and wheat straw was undertaken to investigate the the effect of mycotoxin contamination on dry matter and organic matter digestibilities. The data revealed a negative effect of most studied mycotoxins on the materials digestibility. Among the investigated mycotoxins, penicillic acid with its two concentrations (5 and 10 nmol) was the most negative, affecting digestibilities of both feed materials. Wheat straw digestibility was more influenced than berseem hay by the ochratoxin A, citrinin and sterigmatocystin (besides the penicillic acid) particularly with their high level (10 nmol). Yet, some mycotoxins act as antibiotics which may affect only the harmful flora but encourage the rumen microflora resulting in slight improvement of digestibility. The rumen conditions were able to metabolize or deform the used levels of all mycotoxins studied. Thus, there were no detectable residues of these mycotoxins in the digestion media after the in vitro fermentation.

Aflatoxins↗

RNA silencing of mycotoxin production in Aspergillus and Fusarium species.

Mycotoxins are natural fungal products that are defined by their harmful effects on humans and animals. Aflatoxin contamination of maize by Aspergillus species and trichothecene contamination of small grains by Fusarium species are two of the most severe mycotoxin problems in the United States. We are investigating RNA silencing in an effort to identify novel ways to control mycotoxin contamination of crops. Transformation of two Aspergilli (A. flavus and A. parasiticus) and a Fusarium (F. graminearum) with inverted repeat transgenes (IRT) containing sequences of mycotoxin-specific regulatory genes suppressed mycotoxin production in all three plant-pathogenic fungi. This atoxigenic phenotype was stable during infection on corn and wheat, and importantly, F. graminearum IRT strains were less virulent on wheat than were wild type. The IRT did not alter physiological characteristics of the fungi, such as spore production and growth rate on solid media. These results indicate that RNA silencing exists in Aspergillus and Fusarium plant pathogens and suggest that RNA silencing technology may be a useful tool for eliminating mycotoxin contamination of agricultural products.

Aspergillus↗

Genotoxicity assessment of five tremorgenic mycotoxins (fumitremorgen B, paxilline, penitrem A, verruculogen, and verrucosidin) produced by molds isolated from fermented meats.

A number of toxinogenic fungal species, particularly producers of tremorgenic mycotoxins, have been isolated from traditional fermented meats. Tremorgenic mycotoxins are a group of fungal metabolites known to act on the central nervous system, causing sustained tremors, convulsions, and death in animals. However, the mode of action of these mycotoxins has not been elucidated in detail, and their genotoxic capacity has hardly been investigated. Because genotoxicity is one of the most prominent toxicological end points in food safety testing, we assessed the genotoxicity of five tremorgenic mycotoxins (fumitremorgen B, paxilline, penitrem A, verrucosidin, and verruculogen) associated with molds found in fermented meats. The mycotoxins were tested in two short-term in vitro assays with the use of different genotoxic end points in different phylogenetic systems (the Ames Salmonella/mammalian-microsome assay and the single-cell gel electrophoresis assay of human lymphocytes). According to the results obtained in this study, all of the investigated mycotoxins except penitrem A exhibited a certain degree of genotoxicity. Verrucosidin appeared to have the highest toxic potential, testing positive in both assays. Verruculogen tested positive in the Salmonella/mammalian-microsome assay, and paxilline and fumitremorgen B caused DNA damage in human lymphocytes. The use of fungal starter cultures to avoid tremorgen contamination in fermented meats is recommended.

DNA Damage↗

Airborne fungal ecological niche determination as one of the possibilities for indirect mycotoxin risk assessment in indoor air.

Based on the microbiological analysis of air samples from occupied spaces, some possibilities for indirect risk assessment of mycotoxin-related health problems are proposed. Airborne fungi could be classified on the basis of the relationship between the two environmental factors and their combinations, i.e., temperature and water requirements (water activity aw). One type involves three different groups of molds, selected on the basis of the quantitative and qualitative information about the ability of fungi to sporulate under different environmental conditions: group (i), represented by Aspergillus nidulans, A. niger, and A. ochraceus, and characterized by sporulation which was more dependent on temperature than on water activity; (ii), represented by A. flavus and A. versicolor, in which sporulation was approximately equal and depended on both the temperature changes and aw alterations; and (iii), represented by Cladosporium sp., Penicillium cyclopium, and P. citrinum, in which sporulation depended more on alteration of the aw conditions than on temperature changes. Another type is characterized by four sporulation rates with two levels of mycotoxin risk accumulation in the spores (conidia) of each mold species: large (Ia) and moderate (Ib) sporulation rates with a risk of mycotoxin accumulation (aw > or = 86; t > or = 12 degrees C); rare sporulation (IIa) and absence of sporulation (IIb), without risk of mycotoxin accumulation (aw < or = 86; t < or = 12 degrees C). In conclusion, providing a useful guide for two dimensions, temperature and water activity for each of the three phases of fungal growth, i.e. germination, growth, and sporulation, could be important for determination of the fundamental niche of each fungus and its ability to form or accumulate mycotoxin. Special emphasis should be given to the indirect mycotoxin risk assessment in heating, ventilation, and air conditioning systems.

Aflatoxin B1↗