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PubMed1978

Neurobehavioral studies of tremorgenic mycotoxins verruculogen and penitrem A.

Verruculogen (V) and penitrem A (PA) represent a group of toxic secondary metabolites of mold contaminants of foodstuffs known as 'tremorgenic mycotoxins', which produce a unique neurotoxic syndrome characterized by sustained tremors, limb weakness, ataxia and convulsions. In the present study the intraperitoneal median tremogenic dose in mice for V was found to be 0.92 mg/kg and that for PA, 0.19 mg/kg. Behavioral and neurochemical parameters were assessed following acute exposure to varying neurotoxic and subneurotoxic doses of these mycotoxins. Measures of spontaneous motor activity (photoactometer) and exploratory reflex behaviors (open field) were markedly depressed by both V and PA. Notably, at dose levels of V or PA that were not accompanied by any overt signs of neurotoxicity, significant neuromotor depression was still observed. Acquisition of a conditioned avoidance shuttle response was disrupted, but only at a high neurotoxic dose level of V. Neurochemical analyses revealed no clear catecholaminergic or cholinergic involvement in the neurotoxic syndrome of eigher V or PA.

Acetylcholinesterase↗

PubMed2004

Mycotoxins in root extracts of American and Asian ginseng bind estrogen receptors alpha and beta.

The estrogenic activity of ginseng has been the subject of conflicting reports. Cell proliferation, induction of estrogen-responsive genes, and isolated cases of adverse reactions such as postmenopausal vaginal bleeding and gynecomastia have been reported after ginseng treatment. Other studies report antiproliferative effects with no induction of estrogen-responsive genes. We developed estrogen receptor (ER) alpha and ER alpha competitive binding assays using recombinant receptors and [(3)H]-17 alpha-estradiol to detect phytoestrogens in extracts of Asian ginseng root (Panax ginseng C. A. Meyer) and American ginseng root (Panax quinquefolius L.). Root extracts contained substances that bound both receptor isoforms. These substances had a two to three times greater affinity for ER alpha. Significantly higher binding was found in methanol extracts than in hot water extracts. Subsequent analysis of the extracts revealed significant ER binding attributable to zearalenone, the estrogenic mycotoxin produced by several Fusarium species. The ER showed no binding affinity for Rb1 and Rg1, the major ginsenosides found in P. quinquefolius and P. ginseng, respectively. Thus, ginseng extraction methods, plant species tested, and mycotoxin contaminants may help to explain the disparate literature reports. The prevalence and health significance of fungal contamination in herbal products used for medicinal purposes should be further investigated.

Binding, Competitive↗

PubMed2006

Induction of apoptosis by disturbing mitochondrial-membrane potential and cleaving PARP in Jurkat T cells through treatment with acetoxyscirpenol mycotoxins.

Paecilomyces tenuipes is a famous Chinese medicinal entomopathogenic fungus that grows within the larvae of silkworms. 4beta-acetoxyscirpendiol (4-MAS), a cytotoxic compound belonging to the scirpenol subfamily of trichothecene mycotoxin, was isolated from Paecilomyces tenuipes. To further elucidate the cytotoxic mechanism of 4-MAS, evidences of its induction of apoptosis, together with the structurally related acetoxyscirpenol moiety mycotoxins (ASMs) such as, 15-acetoxyscirpenol (15-MAS), 4,15-diacetoxyscirpenol (4,15-DAS), and 3alpha-acetyldiacetoxyscirpenol (TAS), in the human Jurkat T cell line were reported herein. In the MTT reduction and time-course cytotoxicity assays for monitoring cell viability, all the four ASMs that were tested exhibited cytotoxicity; single acetoxylation at C-4 of the scirpenol family resulted in relatively weak cytotoxicity, while acetoxylation at C-15 resulted in strong cytotoxicity regardless of the other acetoxylations at the C-3 and/or C-4 positions. Phosphatidylserine externalization was induced by all the ASMs that were treated at an early phase in a time-dependent manner, showing a typical apoptotic phenomenon, not a necrotic one. The ASMs also reduced the mitochondria's inner-membrane potential (deltaPsim) through flow cytometry analysis after staining these with DiOC6, a mitochondria-specific and voltage-dependent dye. Acetoxylation of ASM at C-15 increased deltaPsim disruption, but that at C-3 reduced the deltaPsim. The ASMs that were tested also cleaved 113 kDa PARP to an 89-kDa fragment through Western blot assay, suggesting the activation of caspase-3 and/or caspase-7 in the Jurkat T cell. DNA fragmentation was also observed to have been increased in a time-dependent manner by the ASMs that were tested in Jurkat T cells, resulting in the DNA fragmentation intensity order of 4,15-DAS>15-MAS>TAS>4-MAS. These data indicate that the Jurkat T cells that were treated with ASMs underwent typical cascades of apoptotic cell death.

Animals↗

PubMed2003

Expression in cereal plants of genes that inactivate Fusarium mycotoxins.

Trichothecene 3-O-acetyltransferase (encoded by Tri101) inactivates the virulence factor of the cereal pathogen Fusarium graminearum. Zearalenone hydrolase (encoded by zhd101) detoxifies the oestrogenic mycotoxin produced by the same pathogen. These genes were introduced into a model monocotyledon rice plant to evaluate their usefulness for decontamination of mycotoxins. The strong and constitutive rice Act1 promoter did not cause accumulation of TRI101 protein in transgenic rice plants. In contrast, the same promoter was suitable for transgenic production of ZHD101 protein; so far, five promising T0 plants have been generated. Low transgenic expression of Tri101 was suggested to be increased by addition of an omega enhancer sequence upstream of the start codon.

Acetyltransferases↗

PubMed1985

Aflatrem: a tremorgenic mycotoxin with acute neurotoxic effects.

Tremorgenic mycotoxins induce neurologic symptoms ranging from mental confusion to tremors, seizures and death, and are apparently the only class of mycotoxins with significant central nervous system activity. Tremorgens have been implicated in a number of neurologic diseases of cattle collectively known as staggers syndromes, and pose significant agricultural and health problems for both cattle and humans. Although the effects of tremorgens are thought to result from transient perturbations of amino acid neurotransmitter release mechanisms, there is reason to believe that acute exposures to toxins with such synaptic effects may result in degeneration of neuronal fiber processes. To test this hypothesis, rats were given a single tremorgenic (3 mg/kg, IP) dose of aflatrem, and kinetics of amino acid neurotransmitter uptake was assessed in isolated hippocampal nerve terminals at 1 day, 1 week, and 2 weeks after injection. Results indicate a decrease in the capacity of the GABA and glutamate uptake systems, which was interpreted as a loss of nerve terminals. The affinity constants suggest a decrease in release of these transmitters as well. In addition to its transient influence on transmitter release, a single low dose of aflatrem is able to induce degeneration of neuronal processes in hippocampal neurotransmitter systems and therefore represents a long-term health threat.

Animals↗

PubMed1986

Toxicity of mycotoxins for the rat pulmonary macrophage in vitro.

The presence of mycotoxins in grains is well documented. Workers in grain handling occupations are commonly exposed to grain dust aerosols. Work in our laboratory has shown that T-2 toxin is highly toxic to rat alveolar macrophages in vitro, causing loss of viability, release of radiolabeled chromium, inhibition of macromolecular synthesis, inhibition of phagocytosis, and inhibition of macrophage activation. Similarly, patulin caused a significant release of radiolabeled chromium, decrease in ATP levels, significant inhibition of protein and RNA synthesis, and inhibition of phagocytosis. The data show that both T-2 toxin and patulin are highly toxic to rat alveolar macrophages in vitro. The data further suggest that the presence of these mycotoxins in airborne respirable dust might present a hazard to exposed workers.

Animals↗

PubMed1977

The mycotoxins and human health hazards.

The general background of the role of mycotoxins in human disease is briefly reviewed. The acute effects of the ingestion of the aflatoxins and the role that long term exposure may play in human disease are examined in detail. The necessity for co-operation between chemists, veterinarians, physicians and mycologists in the elucidation of the role of the mycotoxins in human disease is stressed.

Acute Disease↗

PubMed2002

The immunosuppressive effect of Fusarium mycotoxin as a function of HLA antigens.

We examined the blastogenic response to phytohaemaglutinin (PHA) in HLA-B8, DR3 positive and negative subjects in the presence or absence of the immunosuppressive Fusarium mycotoxin. HLA-B8, DR3 haplotype was associated with a depression of the response to mitogen in the absence of the mycotoxin, whereas in the presence of deoxynivalenol we could not detect significant differences among individuals either possessing or lacking this haplotype.

Fusarium↗

PubMed2001

Biotic and abiotic factors limiting efficacy of Bt corn in indirectly reducing mycotoxin levels in commercial fields.

Incidence of insect damage, and association of insect damage with mycotoxigenic corn ear molds and mycotoxins was examined in commercial fields of Bt and non-Bt hybrids of different backgrounds in Illinois in 1998 and 1999. Nearly 50% Helicoverpa zea (Boddie) infestation sometimes occurred in Bt hybrids that express high levels of the protein in silks and kernels. Damage by European corn borer, Ostrinia nubilalis Hübner, was uncommon, even in non-Bt ears. Levels of total fumonisins were generally less (15- to 1.8-fold) in Bt versus non-Bt hybrids at the same site, with some significant differences. There were several instances where there were no significant differences in fumonisin levels between low/no Bt kernel hybrids and Bt hybrids that produced high levels of the protein in the kernel and silk tissue. However, significant correlations were often noted between numbers of insect-damaged kernels and total fumonisin levels, especially in 1998, suggesting in these cases that reducing insect damage was still reducing fumonisin levels. There was variability between the correlation coefficient for numbers of insect damaged kernels and fumonisin levels at different sites for the same year, different hybrids at the same site, and the same hybrid for different years. Although reductions in fumonisins in Bt hybrids were more limited than reported in the past, planting the Bt hybrids still appears to be a useful method for indirectly reducing mycotoxins in corn ears.

Aflatoxin B1↗

PubMedDate not supplied

Evaluation of two mycotoxin binders to reduce toxicity of broiler diets containing ochratoxin A and T-2 toxin contaminated grain.

In order to assess ochratoxin A (OA) and T-2 toxin (T-2) binding ability of two commercial sorbents, both in vitro and in vivo trials with broilers were performed. Crude OA and T-2 extracts from contaminated grain were used to assess in vitro binding ability of two sorbents (Zeotek [Zk] and Mycofix [Mx]), by quantifying free mycotoxin through an enzyme-linked immunosorbent assay (ELISA) test. For in vivo trial, a 3 x 2 x 2 factorial arrangement was used for this experiment, being the factors: adsorbents (none, Zk, and Mx), OA (0 and 567 parts per billion [ppb]) and T-2 (0 and 927 ppb). OA and T-2 contaminated wheat and corn, respectively, were added to sorghum-soybean meal diets to meet 567 ppb of OA and 927 ppb of T-2. Mycotoxins were fed alone or combined in treatments. After 21 days, blood chemistry, gross, and histological evaluations were performed. Relative weights of liver, kidney, and bursa of Fabricius were obtained. Zk had the highest OA and T-2 in vitro binding ability (100% and 8.67%, respectively). Chickens fed OA with or without sorbents had a lower body weight and feed intake reduction. However, those birds fed T-2 were partly protected by a sorbent. Birds fed both toxins showed toxic additive effects, and no protection of any adsorbent was observed. A significant reduction in plasma proteins, albumin, and globulins was a characteristic observed in all birds fed diets with OA both with or without adsorbents. Uric acid level in blood was increased in all chickens fed OA-contaminated diets. Histological findings observed in birds fed OA-contaminated diets were necrosis of kidney tubular cells, swollen and necrotic hepatocytes, bile ducts hyperplasia, and increased diameter of proventriculus glands. In birds that received T-2 alone, only the liver, with the same kind of lesions, was affected. According to these results, it can be concluded that there is not a relation between in vitro and in vivo trials. OA toxic effects could not be counteracted by any sorbent. T-2 toxicity could be partially counteracted by an adsorbent used in this research.

Adsorption↗

PubMed1993

Analysis of naturally occurring mycotoxins in feedstuffs and food.

Aflatoxins, zearalenone, deoxynivalenol, fumonisins, and their respective metabolites require specific procedures for their determination because of their diverse chemistry and occurrence in complex matrices of feedstuffs and foods. Major sources of error in the analysis of these mycotoxins arise from inadequate sampling and inefficient extraction and cleanup procedures. The determinative step in the assay for each of these toxins is sensitive to levels below those that are considered detrimental to humans and animals. Aflatoxins can be determined in grains and animal fluids and tissues by TLC, HPLC, gas chromatography-mass spectrometry (GC-MS), and ELISA procedures. Zearalenone, an estrogenic mycotoxin, can readily be determined in cereal grains and foods by HPLC (50 ng/g) and by TLC (300 ng/g). No incurred levels of zearalenone or its metabolites have been detected in animal tissues destined for human consumption. Deoxynivalenol can be determined in wheat and corn at 300 ng/g by a rapid TLC procedure and at 325 ng/g by a GC method. Although not tested collaboratively, an HPLC procedure and an ELISA screening procedure are capable of detecting deoxynivalenol at low (nanograms/gram) levels in feedstuffs and foods. The recently characterized fumonisins can be detected by TLC, HPLC, and GC-MS at levels below those now considered harmful. Thin-layer chromatography and HPLC (with fluorescence detection of derivatives) procedures can detect fumonisins at approximately 100 ng/g; GC-MS is required for detection at lower levels.

Aflatoxins↗

PubMed2005

Effects of feeding a blend of grains naturally contaminated with Fusarium mycotoxins on feed intake, metabolism, and indices of athletic performance of exercised horses.

An experiment was conducted to determine the effect of feeding blends of grains naturally contaminated with Fusarium mycotoxins to mature, exercised horses, and to test the efficacy of a polymeric glucomannan mycotoxin adsorbent (GM polymer) in preventing Fusarium mycotoxicoses. Six mature, mixed-breed mares with an average BW of 530 kg were assigned to one of three dietary treatments for 21 d in a replicated 3 x 3 Latin square design. Feed consumed each day was a combination of up to 3.5 kg of concentrates and 5.0 kg of mixed timothy/alfalfa hay (as-fed basis). The concentrates fed included 1) manage; 2) blend of contaminated grains; and 3) contaminated grains + 0.2% GM polymer (MTB-100, Alltech Inc., Nicholasville, KY). Concentrates containing contaminated grains averaged 11.0 ppm deoxynivalenol, 0.7 ppm 15-acetyldeoxynivalenol, and 0.8 ppm zearalenone (as-fed basis). Feed intake and BW were monitored over a 21-d period. Horses were maintained on a fixed exercise schedule throughout the experiment. At the end of the experiment, each horse completed a time-to-fatigue treadmill step test. Variables measured during pretest, each step of the test, and 5 and 10 min posttest were as follows: 1) time-to-fatigue, 2) heart rate, 3) hematological variables, and 4) serum lactate concentration. Each step consisted of 2 min of fast trot with a 2% increase in incline after each 2 min. Feed intake by horses fed contaminated grains was decreased compared with controls throughout the experiment (P < 0.05). Supplementation of 0.2% GM polymer to the contaminated diet did not alter feed intake by horses compared with those fed the unsupplemented contaminated diet. All hay was consumed regardless of concentrate fed. Weight loss from 0 to 21 d was observed in horses fed contaminated grains compared with controls (P < 0.05). No effect of diet was seen on variables used to measure athletic ability, although the results showed an expected response to exercise for a fit horse. We conclude that exercised horses are susceptible to Fusarium mycotoxicoses as indicated by appetite suppression and weight loss.

Animal Feed↗

PubMed1990

Action of a mycotoxin (diacetoxyscirpenol) on the immune response of the mouse--interaction with an immunomodulator (OM-89).

The action of diacetoxyscirpenol (DAS)--a mycotoxin that belongs to the family of trichothecenes--on the immune system of the mouse was investigated. Two experimental models were used: 1) bacterial infection with Salmonella typhimurium and 2) the PFC (plaque-forming cells) test on the splenic lymphocytes of the mouse. The results obtained showed that these were dependent on the chronological order of the administration of DAS. When the toxin was administered after a bacterial infection or antigenic stimulation, it was observed an increase in the mortality rate and a very significant decrease in the antibody response. By contrast, when this mycotoxin was administered before the bacterial infection or antigenic stimulation, the results obtained were close to those from the controls. The importance of the administration of an immunomodulator of bacterial origin OM-89 before the immunodepression with DAS was shown.

Adjuvants, Immunologic↗

PubMed1991

Mouth lesions in broiler chickens caused by scirpenol mycotoxins.

Dietary scirpentriol (STO), triacetoxyscirpenol (TAS), monoacetoxyscirpenol (MAS), and diacetoxyscirpenol (DAS), mycotoxins produced by Fusarium species, were compared for their ability to cause mouth lesions when graded dietary levels (0, 1, 2, 4, and 8 micrograms STO or TAS/g; 0, .5, 1, 2, and 4 micrograms MAS or DAS/g) were fed to male broiler chickens for 21 days after hatching. The mouth lesions provoked by each scirpenol were dose-related. The minimum effective doses (MED) were 4, 2, 1, and .5 micrograms/g for TAS, STO, DAS, and MAS, respectively, whether the number of affected birds or the number of affected mouth parts (angles, upper beak, lower beak, and tongue) was the measured response. Lesion sites in the mouth varied with the toxin. The rank orders from greatest to least affected sites were angles, upper beak, lower beak, and tongue for TAS and STO, upper beak, lower beak, angles, and tongue for MAS, and upper beak, lower beak, tongue, and angles for DAS. Mouth lesions were clearly visible with each toxin after feeding for 1 wk and the numbers of affected mouth parts almost tripled after 2 wk exposure. During Week 3 of exposure, only the increase caused by MAS was significant (P less than .05). The MED for growth inhibition were 2, 2, 2, and 8 micrograms/g for STO, MAS, DAS, and TAS, respectively. Thus, mouth lesions were of equal or greater sensitivity than growth inhibition as an indicator of scirpenol toxicity. It would appear that the discovery of mouth lesions in birds justifies a mold and mycotoxin control program.

Acetylation↗

PubMed1992

Abnormal feathering of chicks caused by scirpenol mycotoxins differing in degree of acetylation.

Graded levels of the Fusarium mycotoxins, scirpentriol (STO), 15-monoacetoxyscirpenol (15-MAS), 4,15-diacetoxyscirpenol (4,15-DAS), and 3,4,15-triacetoxyscirpenol (TAS), were fed to chicks until 3 wk of age. The primary wing feathers, which were scored visually on a scale of 1 to 5 using a newly created scoring scheme, were altered in a dose-related fashion by 15-MAS [minimum effective dose (MED) = .5 microgram/g diet], 4,15-DAS (MED = 2 micrograms/g), and STO (MED = 4 micrograms/g) but not by TAS (MED greater than 8 micrograms/g). The minimum growth inhibitory doses were 2 micrograms/g for STO, 15-MAS, and 4,15-DAS and 8 micrograms/g for TAS. The main alteration of the feathers was a frayed and missing web on the medial side of the distal half of the feather. The shafts of the feathers tended to have an accentuated medial curve. These results imply that the feather alterations associated with corn and feed infested with Fusarium spp. might be caused by trichothecene mycotoxins such as the scirpenols.

Acetylation↗

PubMed2001

Dietary strategies to counteract the effects of mycotoxins: a review.

We reviewed various dietary strategies to contain the toxic effects of mycotoxins using antioxidant compounds (selenium, vitamins, provitamins), food components (phenolic compounds, coumarin, chlorophyll and its derivatives, fructose, aspartame), medicinal herbs and plant extracts, and mineral and biological binding agents (hydrated sodium calcium aluminosilicate, bentonites, zeolites, activated carbons, bacteria, and yeast). Available data are primarily from in vitro studies and mainly focus on aflatoxin B1, whereas much less information is available about other mycotoxins. Compounds with antioxidant properties are potentially very efficacious because of their ability to act as superoxide anion scavengers. Interesting results have been obtained by food components contained in coffee, strawberries, tea, pepper, grapes, turmeric, Fava tonka, garlic, cabbage, and onions. Additionally, some medicinal herbs and plant extracts could potentially provide protection against aflatoxin B1 and fumonisin B1. Activated carbons, hydrated sodium calcium aluminosilicate, and bacteria seem to effectively act as binders. We conclude that dietary strategies are the most promising approach to the problem, considering their limited or nil interference in the food production process. Nevertheless, a great research effort is necessary to verify the in vivo detoxification ability of the purposed agents, their mode of action, possible long-term drawbacks of these detoxification-decontamination procedures, and their economical and technical feasibility.

Animals↗

PubMed2001

Production of alternaria mycotoxins by Alternaria alternata isolated from weather-damaged wheat.

The production of Alternaria mycotoxins by Alternaria alternata isolated from Chinese weathered wheat kernels were first investigated on polished rice and durum wheat grains. These mycotoxins included alternariol (AOH) and its monomethyl ether (AME), altenuene (ALT), altertoxin I (ATX-I), and tenuazonic acid (TA). Of 25 isolates tested, all were AOH and AME producers, 21 (84%) coproduced ALT and ATX-I, and 8 (32%) produced TA in rice culture. TA was the most abundant toxin produced at a level ranging from 1,369 to 3,563 mg/kg. Much smaller amounts of AOH, AME, ALT, and ATX-I were present with average concentrations of 54, 40, 44, and 8 mg/kg, respectively. There were linear correlations between the level of AOH and AME (r = 0.846), alternariols (AOH plus AME) and ALT (r = 0.785), and ATX-I and TA (r = 0.553). Polished rice medium seems to support a bit more production of Alternaria metabolites than wheat but with an insignificant difference in concentrations (P > 0.05). A study of the time-course of toxin production by A. alternata isolates indicated that AOH production began faster than any other toxins monitored, and ALT production exhibited a progressive increase throughout the experiment. TA producers might reveal their considerably higher ability to produce toxin in the field despite their low frequency.

Alternaria↗

PubMed2005

Occurrence of mycotoxin in Farro samples from southern Italy.

The occurrence of nine mycotoxins and of contamination by pre- and postharvest fungal pathogens of cereals was investigated in samples of stored Triticum monococcum L., Triticum dicoccon Schrank (emmer), and Triticum spelta L. (spelt). In Italy, all three species are collectively referred to as farro. The samples examined were harvested in summer 2000 from eight different sites in southern Italy. Conventional fluorimetric and diode array-based high-performance liquid chromatography (HPLC) analyses and HPLC-mass spectrometry analyses were used to identify fumonisin B1 in five samples (up to 70.00 microg/ kg), ochratoxin A in seven samples (up to 4.07 microg/kg), and beauvericin in three samples (up to 4.44 mg/kg). Enniatin B was detected in one sample (30.00 microg/kg), but no zearalenone or fusaproliferin was found. Deoxynivalenol and aflatoxins were not evaluated. The potentially mycotoxigenic fungal species detected were Alternaria alternata, Fusarium proliferatum, Fusarium tricinctum, Penicillium verrucosum, and Penicillium chrysogenum. This is the first report of the natural occurrence of mycotoxins in farro samples.

Chromatography, High Pressure Liquid↗