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The role of Stachybotrys mycotoxins in building-related illness.

Recently there has been increased attention among both the public and health professionals regarding the potential role of mycotoxins, primarily from fungi of the genus Stachybotrys, as etiologic agents related to illness among persons exposed in the indoor (nonindustrial) environment. Recommendations for the remediation of buildings are being made based in part on reported health effects believed to be due to mycotoxins. A search of NIOSHTIC (a literature database maintained by the National Institute for Occupational Safety and Health) and MEDLINE (from 1965 to present) for literature related to fungi, mycotoxins, and the indoor environment was conducted. References from relevant articles also were reviewed. This strategy yielded a total of 13 articles. Important issues concerning exposure assessment and case definitions are inadequately addressed in the literature reviewed, making it difficult to implicate mycotoxins as a cause of building-related illness. The literature review indicates that currently there is inadequate evidence supporting a causal relationship between symptoms or illness among building occupants and exposure to mycotoxins. Research involving the identification and isolation of specific fungal toxins in the environment and in humans is needed before a more definitive link between health outcomes and mycotoxins can be made.

Air Pollution, Indoor↗

Antibodies against molds and mycotoxins following exposure to toxigenic fungi in a water-damaged building.

Exposure to molds in water-damaged buildings can cause allergy, asthma, hypersensitivity pneumonitis, mucus membrane irritation, and toxicity--alone or in combination. Despite this, significant emphasis has been placed only on Type I allergy and asthma, but not on the other 3 types of allergies. In this study, we sought to evaluate simultaneous measurements of immunoglobulin (Ig) G, IgM, IgA, and IgE antibodies against the most common molds, and their mycotoxins, cultured from water-damaged buildings. Antibodies against 7 different molds and 2 mycotoxins were determined by enzyme-linked immunosorbent assay (ELISA) in the blood of 40 controls and 40 mold-exposed patients. The IgG antibody levels against all 7 of the molds used, as well as the 2 mycotoxins, were significantly greater in patients than in controls. The IgM antibody levels were significantly different in patients for only 6 of 9 determinations. Regarding IgA determinations, antibodies were elevated significantly against all antigens tested, except Epicoccum. However, the differences in IgE levels in controls and mold-exposed patients were significant only for Aspergillus and satratoxin. These differences implied that, overall, the healthy control group was different from the mold-exposed patients for IgG, IgM, and IgA antibodies, but not for the IgE anti-mold antibody. Most patients with high levels of antibodies against various mold antigens also exhibited elevated antibodies against purified mycotoxins, indicating that the patients had been exposed to mold spores and mycotoxins. Detection of high levels (colony-forming units per cubic meter) of molds--which, in this study, strongly suggested that there existed a reservoir of spores in the building at the time of sampling--along with a significant elevation in IgG, IgM, or IgA antibodies against molds and mycotoxins, could be used in future epidemiologic investigations of fungal exposure. In addition to IgE, measurements of IgG, IgM, and IgA antibodies should be considered in mold-exposed individuals.

Adult↗

Efficacy of a commercial mycotoxin binder in alleviating effects of ochratoxin A, fumonisin B1, moniliformin and zearalenone in adult mink.

The addition of nutritionally inert adsorbents to mycotoxin-contaminated animal feed has been a popular approach to decreasing toxicity in animals and carryover of mycotoxins from contaminated feed to animal by-products. Some studies suggest that esterified glucomannan derived from the cell wall of Saccharomyces cerevisiae is effective in reducing the bioavailability of at least some of the mycotoxins occurring in contaminated feed. Because cereal grains are important components of ranch mink diets, mycotoxicoses in mink is a potential problem faced by mink ranchers. We conducted a series of studies to determine if inclusion of a commercially available esterified glucomannan in ranch mink feed was effective in alleviating clinical signs indicative of exposure to ochratoxin A, fumonisin B1, moniliformin or zearalenone in adult mink. In 4 separate trials, mink were fed diets that contained 2.5, 5 or 10 mg ochratoxin A/kg feed, 200 mg fumonisin B1/kg feed, 20 mg moniliformin/kg feed, or 30 mg zearalenone/kg feed with or without 2 g esterified glucomannan/kg feed. Male mink fed diets containing ochratoxin A had significantly decreased feed intake as well as renal lesions characteristic of exposure to that mycotoxin. Inclusion of the esterified glucomannan did not ameliorate these effects. Male mink exposed to fumonisin B1 had increased urinary sphinganine concentration, which was not significantly reduced by the mycotoxin adsorbent. Male mink that consumed monilformin-contaminated diets had characteristic ultrastructural changes in the heart that were not reduced in severity by the esterified glucomannan. Female mink exposed to zearalenone had increased uterine weight, which was not reversed by inclusion of commercial mycotoxin binder in the contaminated feed. The results of this study suggest that a commercial esterified glucomannan was generally ineffective in alleviating effects indicative of exposure to ochratoxin A, fumonisin B1, monilformin and zearalenone in mink.

Adsorption↗

Limits and regulations for mycotoxins in food and feed.

Mycotoxins are natural contaminants whose presence in food- and feedstuffs cannot be completely avoided. Since several mycotoxins have been associated with and implicated in human and animal diseases there is a need to establish maximum levels, guidelines or action levels for them in some kinds of commodities. International and government authorities in many countries have been investing in mycotoxins research and initiating administrative actions for elaboration of legislation and implementing regulatory measures for the control of mycotoxins. Codex Alimentarius Commission is established international legislation on food and feed. In European Union specific limits and regulations for mycotoxins and other contaminants are constructed under the general Codex standards and based on proposal from European Commission. The legal basis for European Commission became available with the framework Council Regulation (EEC) No 315/93. In this paper, legislation regarding maximum levels for certain mycotoxins in food- and feedstuffs in European Community and other countries were reviewed and discussed.

Animal Feed↗

Methods for determining ochratoxin A and other nephrotoxic mycotoxins.

Chemical assays are of major importance for the determination of mycotoxins. Generally, all chemical methods for the analysis of mycotoxins include the basic steps of extraction, clean-up, separation, detection, quantification and confirmation of identity. The various approaches that exist for the determination of nephrotoxic mycotoxins, and in particular the ochratoxins, are discussed below. In conventional procedures, clean-up is usually achieved by liquid-liquid extraction or adsorption column chromatography, followed by thin-layer chromatography or high-performance liquid chromatography and ultraviolet or fluorescence detection. The recent introduction of methods based on immunochemical principles has had a large impact on analytical methodology for mycotoxins, including the ochratoxins, in both the purification and determination steps. The enzyme-linked immunosorbent assay approach for screening and (semi-)quantitative determination and the immunoaffinity column approach for rapid clean-up followed by conventional instrumental analysis are rapidly gaining ground. These techniques also offer possibilities for automated systems. To assist analysts in improving their data in mycotoxin research, the Community Bureau of Reference of the European Commission has produced several reference materials for mycotoxins; others, such as a reference material for ochratoxin A in grains, are in development.

Chromatography↗

Structure-function relationships of 12,13-epoxytrichothecene mycotoxins in cell culture: comparison to whole animal lethality.

Nineteen 12,13-epoxytrichothecene mycotoxins were tested for their relative capabilities to inhibit protein synthesis in Vero cells and rat spleen lymphocytes. Although the lymphocytes were generally more sensitive to the mycotoxins, good correlation existed between the relative potencies of the various trichothecenes in the two cell systems. The most potent mycotoxins (T-2, verrucarin A and roridin A) have acetyl side groups on, or a hydrocarbon chain between, carbons 4 and 15 of the basic ring structure. Loss of side groups from either of these positions or an isovaleryl group at carbon 8 resulted in reduced protein synthesis inhibition (T-2 to HT-2, neosolaniol or diacetoxyscirpenol). Any combination of loss from all three positions (T-2 triol, T-2 tetraol, 15-monoacetyl DAS, scirpentriol, fusarenon X and deoxynivalenol) further weakens their effect. Reduction of the hydroxyl groups to hydroxides, forming verrucarol and deoxyverrucarol, reduced their effectiveness by over a thousand-fold compared to the most potent mycotoxins. Addition of side groups resulted in reduced effectiveness only when an acetyl group was added to the carbon 3 position of T-2 (acetyl T-2) and deoxynivalenol (3-acetyl deoxynivalenol) or on substitution of an epoxide across the 9,10 carbons of diacetoxyscirpenol (beta-epoxide DAS). Effects of combining these and other mycotoxins were additive and showed no synergism or competition for binding to the active site. When in vitro effects of the mycotoxins were compared with results from whole animal lethality tests, several of the trichothecenes were weak inhibitors of protein synthesis in vitro but had in vivo toxicities similar to that of T-2 toxin. Thus, the in vitro cell response of a given trichothecene is not always an accurate predictor of toxicity in whole animals.

Animals↗

Mycotoxin T-2 and aflatoxin B1 as immunosuppressors in mice chronically infected with Toxoplasma gondii.

The aim of this study was to determine whether repeated ingestion of mycotoxin T-2 (T2) or aflatoxin B1 (AFL) at low doses could contribute to the activation of toxoplasmosis in experimentally infected mice. Mice were divided into two groups: Control (C) and Infected (I). The cyst-forming Beverley strain of Toxoplasma gondii was used to produce the infection one month before treatment with mycotoxins. Mycotoxins were given intragastrically for a 50-day period. The average weight gain was reduced in the groups treated with mycotoxins. Mice developed specific IgG to T. gondii. Histopathological studies showed severe encephalitis in all groups infected. The number of unruptured and ruptured cysts was established and the severity of the lesions was evaluated, the groups treated with mycotoxins being the most severely affected. Immunohistochemical studies of the brain showed free antigen in tissues surrounding ruptured cysts. It is suggested that low and repeated doses of mycotoxins, necessary to produce a subclinical intoxication, precipitate Toxoplasma cyst rupture and consequently the activation of chronic toxoplasmosis.

Aflatoxin B1↗

Protein translation inhibition by Stachybotrys chartarum conidia with and without the mycotoxin containing polysaccharide matrix.

Recent studies have correlated the presence of Stachybotrys chartarum in structures with SBS. S. chartarum produces mycotoxins that are thought to produce some of the symptoms reported in sick-building syndrome (SBS). The conidia (spores) produced by Stachybotrys species are not commonly found in the air of buildings that have been found to contain significant interior growth of this organism. This could be due in part to the large size of the Stachybotrys spores, or the organism growing in hidden areas such as wall cavities. However, individuals in buildings with significant Stachybotrys growth frequently display symptoms that may be attributed to exposure to the organism's mycotoxins. In addition, Stachybotrys colonies produce a "slime" or polysaccharide (carbohydrate) matrix that coats the hyphae and the spores. The intent of this project was to determine whether the carbohydrate matrix and the mycotoxins embedded in it could be removed from the spores by repeated washings with either aqueous or organic solvents. The results demonstrated that the process of spore washing removed compounds that were toxic in a protein translation assay as compared to spores that were washed with an organic solution, however a correlation between carbohydrate removal during the washing process and the removal of mycotoxins from the spore surface was not observed. These data demonstrated that mycotoxins are not likely to be found exclusively in the carbohydrate matrix of the spores. Therefore, mycotoxin removal from the spore surface can occur without significant loss of polysaccharide. We also showed that toxic substances may be removed from the spore surface with an aqueous solution. These results suggest that satratoxins are soluble in aqueous solutions without being bound to water-soluble moieties, such as the carbohydrate slime matrix.

Animals↗

Effects of feeding blends of grains naturally contaminated with Fusarium mycotoxins on performance, metabolism, hematology, and immunocompetence of ducklings.

Experiments were conducted to determine the effects of feeding grains naturally contaminated with Fusarium mycotoxins on performance, metabolism, hematology, and immune competence of ducklings. Four hundred sixty-four 1-d-old White Pekin male ducklings were fed starter (0 to 2 wk), grower (3 to 4 wk), and finisher (5 to 6 wk) diets formulated with uncontaminated grains, a low level of contaminated grains, a high level of contaminated grains, or the higher level of contaminated grains + 0.2% polymeric glucomannan mycotoxin adsorbent. Body weight gains, feed consumption, and feed efficiency were not affected by diet. However, consumption of contaminated grains decreased plasma calcium concentrations after 2 wk and plasma uric acid concentrations at the 4-wk assessment point. Mean corpuscular hemoglobin concentrations and hematocrit decreased when ducks were fed contaminated grains for 4 or 6 wk, respectively. In contrast, total numbers of white blood cells and lymphocytes increased transiently in birds fed contaminated grains for 4 wk. The antibody response to sheep red blood cells (CD4+ T cell dependent) and the cell-mediated response to phytohemagglutinin-P (also CD4+ T cell dependent) were not affected by diet, but consumption of contaminated grains for 6 wk decreased the duration of peak cell-mediated response to dinitrochlorobenzene (CD8+ T cell dependent) assessed in a skin test. Feeding grains naturally contaminated with Fusarium mycotoxins, even at levels widely regarded as high, exerted only minor adverse effects on plasma chemistry and hematology of ducklings, and production parameters were unaffected in this avian species. Mycotoxin-contaminated feeds may, however, render these animals susceptible to infectious agents such as viruses against which the CD8+ T cell provides necessary defence. Glucomannan mycotoxin adsorbent was not effective in preventing alterations caused by Fusarium mycotoxins.

Aging↗

A sensitive zonagenetic assay for rapid in vitro assessment of estrogenic potency of xenobiotics and mycotoxins.

Mounting evidence confirms that hepatic biosynthetic processes are essential for female sexual maturation in fish, which is directly controlled by estrogens. These oogenetic events (zonagenesis and vitellogenesis) are induced in both sexes by estrogens. In this paper, we report the induction of zona radiata (zr) proteins and vitellogenin in primary hepatocytes from Atlantic salmon (Salmo salar L.) exposed to xenoestrogens and mycotoxins. Cells were treated with doses of 1, 5, and 10 microM 4-nonylphenol (4-NP), o, p'-DDT, lindane ([gamma]-HCH), and bisphenol A (BPA), which all induced zr proteins and vitellogenin in an approximate dose-dependent manner. Hepatocytes were also treated with combinations of xenoestrogens at 1 or 2 microM, resulting in elevated levels of both zr proteins and vitellogenin, compared to single treatment. The estrogenic activity of the mycotoxin zearalenone (ZEA) and its metabolites [alpha]-ZEA) and ss-zearalenol (ss-ZEA)], with regard to zonagenesis and vitellogenesis, was assessed in this assay system. Mycotoxins were used at concentrations of 10, 100, or 1,000 nM. All induced zr proteins and vitellogenin, with [alpha]-ZEA being the strongest inducer. When cells were treated with xenoestrogens or mycotoxins in combination with an estrogen receptor inhibitor (ICI 182,780), the induction of both zr proteins and vitellogenin was inhibited in all cases. Thus, the reported estrogen effects are bonafide estrogen responses. Zona radiata proteins were more responsive than vitellogenin to both xenoestrogens and mycotoxins. The versatility and sensitivity of the hepatocyte assay demonstrates that biosynthesis of zr proteins provides a new supplementary method for estimating xenoestrogenicity and mycotoxin action.

Animals↗

How much should we involve genetic and environmental factors in the risk assessment of mycotoxins in humans?

Despite consented efforts in prevention, mycotoxins remain a problem of human health concern in several parts of the world including developed countries. Within the same range of toxins concentrations in the blood some people develop a disease while others do not. Could this inequality in front of mycotoxins effects be explained by environment factors and/or genetic predisposition? Among recent advances in environmental health research Correlation between chronic diseases and mycotoxins in humans deserves attention through several questions: Are genetic factors involved in disease causation of mycotoxins? How much are these factors currently taken into account for mycotoxins risk assessment and how much should we involve them? Answers are still to come. Genetic and environment factors deserve therefore more attention when dealing with regulatory limits, since among the general population, those who are at risk and will develop specific diseases are likely those bearing genetic predispositions. We have addressed these questions for the specific case of ochratoxin A in humans by investigating in Tunisia, county of Jelma, in four rural families forming a household of 21 persons all exposed to ochratoxin A in diet. Our results confirm that ochratoxin A induces chronic tubular nephropathy in humans and mainly point at those having the HLA haplotype A3, B27/35, DR7 to be more sensitive to the disease for quantitatively similar or lower exposure. Persons with such haplotype were found to bear chronic interstitial nephropathy with tubular karyomegalic cells while others were apparently healthy. Godin et al. (1996) in France have also found in sibling (a sister and her brother from urban area) that have similar HLA haplotype B35-patern, OTA-related renal tubulopathy with mild proteinuria including beta2-microglobulinuria. Several mechanisms are discussed that could be put ahead to explain how the HLA haplotype could lead to tubular cells lyses and renal failure. In the mean time it is urgent to search for mass screening biomarkers for mycotoxins in humans and related genetic factors to set-up more appropriate regulation.

Adult↗

[Mycotoxin producing fungi].

Mycotoxins are relatively small molecules characterized by a diversity of chemical structure and a diversity of biological activity. They are often genotypically specific for a group of species, but the same compound can also be formed by fungi belonging to different genera. Most of the mycotoxins known have been recognized as metabolic products of Aspergillus, Penicillium and Fusarium species. This review will be focused on aflatoxins, ochratoxins and fumonisins because of their hazard to animal and human health. The production of these mycotoxins have been usually associated with a small number of species but some recent studies have reported the production of these mycotoxins by some other species. These results show that mycotoxin production is broader than is normally thought, so the possibility can not be ruled out that new species may be a new source of unexpected mycotoxins in their natural substrates.

English Abstract↗

Inhibitory effect of trichothecene mycotoxins on bovine platelets stimulated by platelet activating factor.

Several species of fungi, which infect cereals and grains, can produce a class of compounds, known as trichothecene mycotoxins, which is characterized by a substituted epoxy-trichothecene ring structure. Cattle are susceptible to intoxication from feeds contaminated with T-2 toxin, one of the more potent trichothecene mycotoxins, while swine refuse to ingest feed contaminated with T-2 toxin. The bovine platelet has been used as a model cell system to evaluate the effects of T-2 toxin and its natural metabolites, HT-2 toxin and T-2 tetraol, on cell function in vitro. Due to the lipophilic nature of these mycotoxins, a biologically active phospholipid was used to stimulate the platelets in the presence and absence of the toxins. The mycotoxin T-2 toxin and its major metabolite HT-2 toxin inhibited platelet activating factor-stimulated bovine platelets, suspended in homologous plasma, in a concentration but not time dependent manner. Significant inhibition of platelet function (p less than 0.01) occurred with 135 ng T-2 toxin per 10(6) platelets and with 77 ng HT-2 toxin per 10(6) platelets. These mycotoxins exerted an additive inhibitory effect on the platelet aggregation response. In contrast, the minor metabolite T-2 tetraol had no inhibitory effect on platelet function and had no influence on the responses of T-2 toxin or HT-2 toxin when the mycotoxins were present together in the platelet suspensions.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effect of the trichothecene mycotoxin diacetoxyscirpenol on nitrosamine-induced oesophageal cancer and on relevant enzymes in oesophagus and liver.

To test the concept that human oesophageal cancer is initiated by nitrosamines and potentiated by consumption of food contaminated by mycotoxins produced by Fusaria, the effect of the trichothecene mycotoxin diacetoxyscirpenol (DS), alone and in combination with N-nitroso-N-methylbenzylamine (NMBzA), on rat oesophagus was studied. Chronic treatment with DS induced oesophageal hyperplasia, but simultaneous treatment with NMBzA tended to inhibit rather than to enhance carcinogenesis. The mycotoxin did not inhibit O6-alkylguanine-DNA alkyltransferase in oesophagus and produced a marked induction of repair protein in liver. Depletion of the repair protein in oesophagus brought about by injection of NMBzA was not inhibited by the mycotoxin, and its reappearance was not delayed. Intubation of DS reduced DNA synthesis in the oesophagus, while dietary treatment resulted in an increase after nine weeks. The results suggest that, while simultaneous treatment with DS reduces cancer induced by NMBzA, if there is first exposure to mycotoxin and induction of hyperplasia, and then exposure to nitrosamine, so that the carcinogen acts on a vulnerable oesophagus in which there is an increased rate of cell proliferation, the mycotoxin could well enhance carcinogenesis.

Animals↗

Fungal drivers of mycotoxin contamination in wheat: Early warning and plasma-based control.

Mycotoxin contamination in wheat is a major food safety concern; however, quantitative evidence linking fungal community signals, mycotoxin exceedance risk, and wheat quality traits in naturally contaminated wheat remains limited. In this study, wheat samples were collected from mycotoxin-prone monitoring sites under unusually rainy conditions in 2022 to explore early-warning indicators and post-harvest mitigation strategies. According to the National Food Safety Standard of China GB 2761-2017, aflatoxin B1 (AFB1), deoxynivalenol (DON), and zearalenone (ZEN) exceeded the maximum limits in 52.24, 47.76, and 23.88% of samples, respectively; 38.81% exceeded the reference EU threshold for T-2 toxin, and 46.27% showed co-contamination with at least two mycotoxins above their respective thresholds. Although Alternaria, Cladosporium, and Epicoccum dominated the fungal community, Fusarium abundance was significantly associated with DON contamination and Fusarium-damaged kernels (FDKs). Mediation analysis identified DON as a significant mediator linking Fusarium abundance to FDKs, accounting for 68.41% of the total effect. In addition, Fusarium abundance above 3.70% showed strong predictive performance for DON exceedance, with an area under the curve of 0.906, indicating its potential as an early-warning indicator. Culture-based assays confirmed the toxigenic potential of Aspergillus and Fusarium isolates under simulated temperature and moisture conditions. After optimization using a toxin-spiked wheat flour model, dielectric barrier discharge cold plasma degraded AFB1, DON, and ZEN by 29.30-35.68%, disrupted the morphology of toxigenic fungi, and did not significantly affect wheat quality. This study provides practical insights into mycotoxin risk warning and post-harvest mitigation in wheat.

Triticum↗

Worldwide regulations for mycotoxins in 1994.

Since the discovery of the aflatoxins in the 1960s, regulations have been established in many countries to protect the consumer from harmful effects of mycotoxins that may contaminate foodstuffs. Various factors play a role in the decision-making process of setting limits for mycotoxins. These include scientific factors such as availability of survey data, toxicological data, analytical methodology, and knowledge about the distribution of mycotoxins in contaminated commodities. Economical and political factors such as commercial interests and sufficiency of food supply have their impact as well. International enquiries on existing mycotoxin regulation in foodstuffs and animal feedstuffs have been carried out several times in the 1980s and details about tolerances, legal bases, responsible authorities, official protocols of analysis and sampling have been published. The most recent enquiry is currently taking place as part of a contract with the Food and Agriculture Organization. At least 77 countries now have regulations for mycotoxins. An overview is presented of the tentative results of this enquiry, including frequency distributions of existing tolerances for aflatoxins and current trends.

Food Analysis↗

Mycotoxins and fermentation--beer production.

Along with food safety issues due to mycotoxins, the effects of Fusarium infections on malt and beer quality can be disastrous. While some of the Fusarium head blight mycotoxins, such as DON, present in infected barley may be lost during steeping, the Fusarium mold is still capable of growth and mycotoxin production during steeping, germination and kilning. Therefore, detoxification of grain before malting may not be practical unless further growth of the mold is also prevented. Methods for reducing the amount of mold growth during malting are needed. Physical, chemical and biological methods exist for inhibiting mold growth in grain. Irradiation is a promising means for preventing Fusarium growth during malting, but its effects on malt quality and mycotoxin production in surviving mold need to be evaluated in more detail. Chemical treatments such as ozonation, which do not leave chemical residues in beer, also appear to be promising. Although biological control methods may be desirable, the effects of these inhibitors on malt and beer quality require further investigation. In addition, storage studies are needed to determine the effect of biological control on Fusarium viability and malt quality. It may also be possible to incorporate detoxifying genes into fermentation yeasts, which would result in detoxification of mycotoxins present in wort. Development of these types of technological interventions should help improve the safety of products, such as beer, made from Fusarium infected grain.

Beer↗

Mycotoxins, mycotoxicoses, mycotoxicology and Mycopathologia.

Mycotoxins are fungal poisons. This definition does not stipulate whether fungi are the targets of poisoning or are the producers of the poisons. The following is suggested as a useful working definition: Mycotoxins are natural products produced by fungi that evoke a toxic response when introduced in low concentration to higher vertebrates and other animals by a natural route. Some mycotoxins have multiple effects, and may cause phytotoxic and antimicrobial syndromes in addition to animal toxicity. By convention, mushroom and yeast poisons are usually excluded from discussions of mycotoxins. The eclectic nature of the discipline and the international scope of the problem has attracted scientists from many different backgrounds. The publishers and editors of Mycopathologia intend for this journal to become a major forum for mycotoxin research.

Animals↗