Microbial transformation of deoxynivalenol (vomitoxin).

Microbial inocula from rumen fluid, soil, and contents of the large intestines of chickens (CLIC) and of swine (SLIC) were tested for their ability to transform deoxynivalenol (vomitoxin) in vitro. Microorganisms in (CLIC) completely transformed pure vomitoxin, and this activity was retained through six serial subcultures. No alteration of the toxin by incubation with SLIC was detected, whereas 35% of the vomitoxin was metabolized in the original culture of rumen fluid and 50% was metabolized by the soil sample, though metabolism was decreased in subsequent subcultures of either sample. A single metabolite was isolated and identified as deepoxy vomitoxin. The increase in concentration of deepoxy vomitoxin in the culture medium corresponded with the decrease in vomitoxin concentration. The vomitoxin transformation rate was not affected by either the ratio of CLIC to vomitoxin (5 to 0.2 g of CLIC per mg of vomitoxin) or the initial concentration of vomitoxin (14 to 1,400 ppm) in the medium. Biotransformation of vomitoxin was completely inhibited when the pH in the medium was lowered to 5.20. Sodium azide at a 0.1% (wt/vol) concentration in the medium blocked the transformation of vomitoxin, suggesting that the deepoxidation of vomitoxin is an energy-dependent process. About 50% of the vomitoxin in moldy corn in culture medium was transformed by microorganisms from CLIC. The vomitoxin transformation rate in moldy corn was not affected when the concentration of CLIC changed from 0.2 to 0.8 g/ml of medium. Vomitoxin in the moldy corn was not transformed when CLIC were added to corn without culture medium.(ABSTRACT TRUNCATED AT 250 WORDS)     

Microbial transformation of deoxynivalenol (vomitoxin).

Microbial inocula from rumen fluid, soil, and contents of the large intestines of chickens (CLIC) and of swine (SLIC) were tested for their ability to transform deoxynivalenol (vomitoxin) in vitro. Microorganisms in (CLIC) completely transformed pure vomitoxin, and this activity was retained through six serial subcultures. No alteration of the toxin by incubation with SLIC was detected, whereas 35% of the vomitoxin was metabolized in the original culture of rumen fluid and 50% was metabolized by the soil sample, though metabolism was decreased in subsequent subcultures of either sample. A single metabolite was isolated and identified as deepoxy vomitoxin. The increase in concentration of deepoxy vomitoxin in the culture medium corresponded with the decrease in vomitoxin concentration. The vomitoxin transformation rate was not affected by either the ratio of CLIC to vomitoxin (5 to 0.2 g of CLIC per mg of vomitoxin) or the initial concentration of vomitoxin (14 to 1,400 ppm) in the medium. Biotransformation of vomitoxin was completely inhibited when the pH in the medium was lowered to 5.20. Sodium azide at a 0.1% (wt/vol) concentration in the medium blocked the transformation of vomitoxin, suggesting that the deepoxidation of vomitoxin is an energy-dependent process. About 50% of the vomitoxin in moldy corn in culture medium was transformed by microorganisms from CLIC. The vomitoxin transformation rate in moldy corn was not affected when the concentration of CLIC changed from 0.2 to 0.8 g/ml of medium. Vomitoxin in the moldy corn was not transformed when CLIC were added to corn without culture medium.(ABSTRACT TRUNCATED AT 250 WORDS)     

Acceptance by swine and rats of corn amended with trichothecenes

Swine and rats demonstrated the same response factor (i.e., the average amount of corn amended with trichothecenes consumed by animals per the average amount of uncontaminated corn consumed by animals) for consumption of corn amended with 40 ppm of either T-2 toxin or diacetoxyscirpenol Rat response factor for corn containing 40 ppm of vomitoxin was 1.8 times more than corn containing either T-2 toxin or diacetoxyscirpenol at 40 ppm. For the corn containing 40 ppm of vomitoxin, swine response factor was 1.8 times greater than rat response factor.     

Acceptance by swine and rats of corn amended with trichothecenes.

Swine and rats demonstrated the same response factor (i.e., the average amount of corn amended with trichothecenes consumed by animals per the average amount of uncontaminated corn consumed by animals) for consumption of corn amended with 40 ppm of either T-2 toxin or diacetoxyscirpenol Rat response factor for corn containing 40 ppm of vomitoxin was 1.8 times more than corn containing either T-2 toxin or diacetoxyscirpenol at 40 ppm. For the corn containing 40 ppm of vomitoxin, swine response factor was 1.8 times greater than rat response factor.     

Immunochemical assessment of mycotoxins in 1989 grain foods: evidence for deoxynivalenol (vomitoxin) contamination.

To assess the potential for mycotoxin contamination of the human food supply following the 1988 U.S. drought, 92 grain food samples were purchased from retail outlets in the summer of 1989 and surveyed for aflatoxin B1, zearalenone, and deoxynivalenol (DON [vomitoxin]) by monoclonal antibody-based competitive enzyme-linked immunosorbent assay (ELISA). Only one sample (buckwheat flour) was found to contain aflatoxin B1 (12 ng/g), whereas zearalenone was found in 26% of the samples at a mean concentration of 19 ng/g. In contrast, the DON ELISA was positive in 50% of the samples at a detection level of 1.0 micrograms/g. Between 63 and 88% of corn cereals, wheat flour/muffin mixes, rice cereals, and corn meal/muffin mixes yielded positive results for DON, whereas 25 to 50% of oat cereals, wheat- and oat-based cookies/crackers, corn chips, popcorn, and mixed-grain cereals were positive for DON. The mean DON content of the positive samples was 4.0 micrograms/g, and the minimum and maximum levels were 1.2 and 19 micrograms/g, respectively. When positive ELISA samples were also analyzed by high-performance liquid chromatography, a strong correlation between the two methods was found. The presence of DON in the two highest samples, corn meal and mixed-grain cereal, which contained 19 and 16 micrograms/g, respectively, was quantitatively confirmed by gas chromatography-mass spectrometry. The results indicated that DON was present in 1989 retail food products at concentrations that exceeded those found in previous market surveys and that have been experimentally associated with impaired animal health.     

Immunochemical assessment of mycotoxins in 1989 grain foods: evidence for deoxynivalenol (vomitoxin) contamination

To assess the potential for mycotoxin contamination of the human food supply following the 1988 U.S. drought, 92 grain food samples were purchased from retail outlets in the summer of 1989 and surveyed for aflatoxin B1, zearalenone, and deoxynivalenol (DON [vomitoxin]) by monoclonal antibody-based competitive enzyme-linked immunosorbent assay (ELISA). Only one sample (buckwheat flour) was found to contain aflatoxin B1 (12 ng/g), whereas zearalenone was found in 26% of the samples at a mean concentration of 19 ng/g. In contrast, the DON ELISA was positive in 50% of the samples at a detection level of 1.0 micrograms/g. Between 63 and 88% of corn cereals, wheat flour/muffin mixes, rice cereals, and corn meal/muffin mixes yielded positive results for DON, whereas 25 to 50% of oat cereals, wheat- and oat-based cookies/crackers, corn chips, popcorn, and mixed-grain cereals were positive for DON. The mean DON content of the positive samples was 4.0 micrograms/g, and the minimum and maximum levels were 1.2 and 19 micrograms/g, respectively. When positive ELISA samples were also analyzed by high-performance liquid chromatography, a strong correlation between the two methods was found. The presence of DON in the two highest samples, corn meal and mixed-grain cereal, which contained 19 and 16 micrograms/g, respectively, was quantitatively confirmed by gas chromatography-mass spectrometry. The results indicated that DON was present in 1989 retail food products at concentrations that exceeded those found in previous market surveys and that have been experimentally associated with impaired animal health.     

Simultaneous occurrence of fumonisin B1 and other mycotoxins in moldy corn collected from the People's Republic of China in regions with high incidences of esophageal cancer.

A total of 31 corn samples collected from households in the counties of Cixian and Linxian of the People's Republic of China, where high incidences of esophageal cancer have been reported, were analyzed for fumonisin B1 (FB1), aflatoxin, and total trichothecene mycotoxins. High levels of FB1 (18 to 155 ppm; mean, 74 ppm) were found in 16 of the samples that showed heavy mold contamination. FB1, at lower levels (20 to 60 ppm; mean, 35.3 ppm), was also found in 15 samples, collected from the same households, that did not show any visible mold contamination. The levels of aflatoxin in the samples were low (1 to 38.4 ppb; mean, 8.61 ppb). High levels of total type-A trichothecenes were also found in the moldy corn samples (139 to 2,030 ppb; mean, 627 ppb). Immunochromatography of selected samples revealed that these samples contained T-2 toxin, HT-2 toxin, iso-neosolaniol, monoacetoxyscirpenol, and several other type-A trichothecenes. The concentration of total type-B trichothecenes in 15 moldy corn samples was in the range of 470 to 5,826 ppb (mean, 2,359 ppb). High levels (3.7 to 5.0 mg/g) of FB1 were produced in corn in the laboratory by five Fusarium moniliforme strains isolated from the moldy corn. These fungi were also capable of forming various nitrosamines (5 to 16 micrograms per flask) in the presence of nitrate and precursor amines.(ABSTRACT TRUNCATED AT 250 WORDS)     

Aspergillus terreus and its toxic metabolites as a food contaminant in some Egyptian Bakery products and grains

A. terreus isolates isolated from some bakery products, corn and rice were found to be able to produce territrems. 90% of theA. terreus isolated from bakery products were able to produce territrem A, with a mean of 0.09 ppm, while 80% ofA. terreus isolates produce territrem B with a mean of 0.24 ppm. On the other hand 31.8% of the isolates ofA. terreus from corn were able to produce territrem A with a mean of 0.44 ppm. ConcerningA. terreus isolates from rice, 66.7% were found to produce territrem A, with a mean of 5.28 ppm, and 77.8% of the isolates produced territrem B with a mean of 1.79 ppm.     

Aflatoxin contamination caused by natural fungal infection of preharvest corn

Summary Aflatoxin contamination of developing corn (Zea mays L.) kernels caused by natural infection byAspergillus flavus Link ex Fries was studied in hybrids developed for the U.S. corn belt and for the southern U.S. and grown at diverse locations in 1977. Planting dates were staggered to examine the effect of crop maturity on infection by the toxin-producing fungus. A broad range of toxin values was observed at harvest; some levels exceeded the highest that had been previously recorded in corn. The highest concentration of aflatoxin B₁ detected was 8030 ppb. Levels of toxin differed significantly among planting dates in Florida and Georgia; the second planting date at these locations contained the highest toxin levels. Elevated concentrations of toxin were characteristic of kernel samples from southern locations and southeast Missouri; at these locations samples from hybrids developed for the south had significantly lower levels of toxin than hybrids developed for the corn belt. Ears with heavy insect damage had higher toxin levels than ears with less evidence of insect attack.Mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

Influence of Phospholipids on Growth, Sporulation and Virulence of the Endoparasitic Fungi Drechmeria coniospora, Verticillium balanoides and Harposporium anguillulae in Liquid Culture

Verticillium balanoides mycelial growth was stimulated on solid corn meal agar (1.7 %) and in liquid corn meal broth (0.2 %) upon the addition of phospholipids at various concentrations. Sporulation differed with phospholipid products and was highest in pure corn meal. Drechmeria coniospora mycelial growth increased upon addition of phospholipids at all concentrations in solid or liquid culture. Sporulation increased at high concentration (1000 ppm) and decreased at low concentration (100 ppm) of phospholipids in the medium. For both fungi, infectivity of conidia produced in liquid culture decreased when compared to conidia from parasitized nematodes. Addition of phospholipids partly restored this effect. Harposporium anguillulae mycelial growth and sporulation was not affected by addition of phospholipids to solid or liquid corn meal medium.     

Survey of 1977 crop year preharvest corn for vomitoxin.

Fifty-two preharvest corn samples were collected in mid-October 1977 from 26 farms in a four-county area of northwest Ohio. Vomitoxin ranging from 0.5 to 10 microgram/g was found in 24 of the samples. Analysis for vomitoxin was by gas-liquid chromatography. Preceding harvest in northwest Ohio, unusual wet conditions prevailed, making it favorable for Fusarium growth. Gibberella zeae-infected kernels ranged from 2 to 50% of the kernels analyzed for 44 corn samples, and 8 corn samples showed no infection.     

Elaboration of vomitoxin and zearalenone by Fusarium isolates and the biological activity of Fusarium-produced toxins.

Sixteen Fusarium isolates belonging to F. graminearium Schw. and F. culmorum (W.G. Smith) Sacc. produced vomitoxin and zearalenone on cracked corn at 28 degrees C. Quantitation for vomitoxin was by gas-liquid chromatography. This toxin was produced in quantities of 5 to 236 microgram/g of fermented corn. Vomitoxin showed weak antibiotic activity against Penicillium digitatum Sacc., Mucor ramannianus Möller, and Saccharomyces bayanus Sacc., but did not inhibit gram-positive, gram-negative, or acid fast bacteria. The two molds and the yeast were inhibited by T-2 toxin at 5 micrograms, and diacetoxyscirpenol inhibited the molds at 5 micrograms and the yeast at 50 micrograms.     

Elaboration of vomitoxin and zearalenone by Fusarium isolates and the biological activity of Fusarium-produced toxins

Sixteen Fusarium isolates belonging to F. graminearium Schw. and F. culmorum (W.G. Smith) Sacc. produced vomitoxin and zearalenone on cracked corn at 28 degrees C. Quantitation for vomitoxin was by gas-liquid chromatography. This toxin was produced in quantities of 5 to 236 microgram/g of fermented corn. Vomitoxin showed weak antibiotic activity against Penicillium digitatum Sacc., Mucor ramannianus M?ller, and Saccharomyces bayanus Sacc., but did not inhibit gram-positive, gram-negative, or acid fast bacteria. The two molds and the yeast were inhibited by T-2 toxin at 5 micrograms, and diacetoxyscirpenol inhibited the molds at 5 micrograms and the yeast at 50 micrograms.     

Suppression of spore germination and aflatoxin biosynthesis in Aspergillus parasiticus during and after exposure to high levels of phosphine

Agar cultures of toxigenic Aspergillus parasiticus NRRL 2999 were exposed to phosphine (PH3), in levels ranging from 0 to 2000 ppm (vol/vol). It was found that with PH3 concentrations of 400 ppm or higher the growth of the fungus was totally arrested. When PH3 was vented and the agar plates were exposed to open air, 100% of the initial CFU developed into fully grown colonies after PH3 levels below 300 ppm, but at higher PH3 concentrations only 50% of the colonies developed. The same strain of A. parasiticus was inoculated into high moisture corn under conditions highly favorable for aflatoxin production, and it was exposed to a range of PH3 levels. After exposure to 500 ppm PH3, both fungal growth and aflatoxin synthesis resumed shortly after elimination of the toxic gas, but after exposure to PH3 levels of 1000 ppm and higher, the physical appearance of the contaminated corn was remarkably changed, showing reduced mycelial growth and almost complete absence of green pigmentation. In addition, aflatoxin synthesis was totally absent for the remainder of the experiment (20 days). These results strongly suggest that exposure to PH3 levels of 1000 ppm or higher could bring about persistent metabolic changes in surviving Aspergillus organisms. This revised version was published online in June 2006 with corrections to the Cover Date.     

Selenium concentrations in crops grown on low-selenium soils as affected by fly-ash amendment

Summary Studies were conducted under greenhouse and field conditions to evaluate the feasibility of using fly-ash amendments to produce selenium-adequate crops (0.10 to 0.30 ppm Se) on low-Se soils without adversely affecting either the crops or the soils. Two soils, a fine-loamy Alfisol belonging to the Honeoye series and a coarse-loamy Inceptisol belonging to the Mardin series were used to make 3 soil treatments thus: Honeoye, limed Mardin and unlimed Mardin. Fly-ash was mixed with the soils at rates of 0, 2500, 5000, 5000, 10000, 25000 and 50000 ppm. In the greenhouse studies the test crops were corn (Zea mays L.), alfalfa (Medicago sativa), birdsfoot trefoil (Loitus corniculatus) and dry beans (Phaseolus vulgaris L.). There were no yield differences due to fly-ash additions. The rates of 25,000 and 50,000 ppm fly-ash produced the desired Se levels in the utilizable parts of the crops. In the field studies, conducted on another fine-loamy Alfisol belonging to the Lima series and using fly-ash at the rates of 0, 11.2, 22.4 and 112.0 metric tons/ha, it was observed that the yields of corn and dry beans grown were also not influenced by the treatments. Increased levels of Se in corn grains (0.10 ppm) and stover (0.08 ppm) and dry bean seeds (0.14 ppm) were observed at the application rate of 112 metric tons per ha.The results indicate that fly-ash has a potential for producing forages and feed crops containing adequate amounts of Se to protect livestock from Se-responsive diseases.     

The inability of Fusarium species extracts to induce dieback in pines

Cultures of F. moniliforme var. subglutinans, F. moniliforme, F. lateritium, F. equiseti, F. semitectum and F. solani from pine and F. moniliforme and F. graminearum from southern U.S. corn were grown on rice and corn, extracted, and checked for toxicity in mice, chicken embryos, and pine seedlings, and for mutagenicity by the Ames test. While extracts from both fungal groups contained toxins, none of the extracts induced dieback in pine seedlings. Almost all of the cultures isolated from corn in contrast to those from pine, were mutagenic. Thin-layer chromatography did not detect T-2 toxin, moniliformin, or vomitoxin, indicating that these toxins do not elicit dieback symptoms in pine.     

Clinical cases of fusarium-toxicoses in Austrian domestic animals in connection with fusariumtoxinrcontaminated feedstuffs

A surveillance study of the most important fusarium-toxins in Austrian feed samples, the estrogenic mycotoxin zearalenone (F-2 toxin) as well as the refusal of feed causing deoxynivalenol (vomitoxin), of the last 8 years is given. The author describes predominantly the biologic effects in swine, cattle and poultry from the practical point of view and the correlation between clinical symptoms of the incriminated animals and the mycotoxin-levels found in the suspicious feed samples. Some field cases, the clinical symptoms and the fusariumtoxin-concentrations found are presented in one table.     

VA-mycorrhizae and mycorrhiza stimulating isoflavonoid compounds reduce plant herbicide injury

Imazaquin, imazethapyr and pendimethalin showed high toxicity to sorghum plants grown in a greenhouse soil mix. However, mycorrhizal sorghum plants were less affected by herbicide toxicity than non-mycorrhizal ones, at low to moderate herbicide concentrations. VAM herbicide safening effects were more evident on imazaquin-treated plants, than for those treated with the other two herbicides. Applications of imazethapyr and pendimethalin at the two highest concentrations, but not imazaquin, reduced VAM colonization rates in sorghum. Applications of the VAM stimulating isoflavonoids, biochanin A and formononetin, at 5 ppm solutions to a field soil sample containing toxic levels of imazaquin (13 ppb) and indigenous VAM fungi, reduced herbicide-induced injury in corn and sorghum under growth chamber conditions. The benefits of isoflavonoids were reduced when additional propagules of Glomus intraradix were added into field-soil samples, and were eliminated when VAM fungi were inactivated by autoclaving. This indicates that herbicide safening effects of biochanin A, and formononetin are VAM-mediated and also suggests the potential use of these isoflavonoids as herbicide safeners.     

A survey of aflatoxin B<Subscript>1</Subscript> and total aflatoxin contamination in baby food,peanut and corn products sold at retail in Indonesia analysed by ELISA and HPLC

Aflatoxin contamination has been well known as a world-wide health-threatening problem in tropical countries including Indonesia. This research was undertaken to determine the degree of aflatoxin contamination in different Indonesian foodstuffs. A preliminary survey was carried out to evaluate the level of total aflatoxin (AfT) and aflatoxin B₁ (AfB₁) contamination of baby foods, peanut products, and corn products, which were purchased from traditional markets and supermarkets in Indonesia during the year 2001-2002. Eighty two peanut products, 12 baby foods products, and 11 corn products from different brands were analysed for AfT and AfB₁ using the Enzyme-Linked Immunosorbent Assay (ELISA) method. The results indicate that, of the brands analysed, 35% of the peanut products were contaminated with aflatoxins at various levels (range 5 to 870 μg/kg). Peanut-chilli sauces had the highest percentage of AfT contamination 9/12 (75%), which was followed by traditional snacks 5/11 (45%), peanut butter 4/11 (40%), flour egg coated peanut 6/16 (37%), and peanut cake 3/10 (30%). Fried peanuts and roasted peanut were found to contain aflatoxin at relatively lower percentages of 9% and 8%, respectively. From the 12 analysed baby food samples, on the other hand, no sample was found to be contaminated with aflatoxins. Two of 11 samples (18%) of corn based products were contaminated with AfT, ranging between 5.8 and 12.4 μg/kg. Additionally, 30 selected samples in different concentration ranges were further analysed to verify the correlation between ELISA and HPLC techniques and results were compared.     

Production of vomitoxin on corn by Fusarium graminearum NRRL 5883 and Fusarium roseum NRRL 6101.

Two vomitoxin-producing isolates of Fusarium spp. were grown on cracked corn for 1 to 8 weeks at 15, 20, 25, 28, and 32 degrees C. Maximum production of vomitoxin by Fusarium graminearum Schw. NRRL 5883 occurred at 30 degrees C and 40 days, and that by Fusarium roseum Schw. NRRL 6101 occurred at 26 degrees C and 41 days. These optimum production points were determined from response surface contour graphs in relation to temperature and time. Only small amounts of vomitoxin were produced at 15 and 20 degrees C by each strain. A 133-microgram quantity of vomitoxin, with an indicated purity of 95%, was isolated per gram of corn fermented with F. graminearum NRRL 5883.