D. Abramson

Comparison of Canadian Fusarium graminearum isolates for aggressiveness, vegetative compatibility, and production of ergosterol and mycotoxins

Fusarium graminearum is the predominant pathogen causing fusarium head blight of cereals in North America. Fifteen Canadian isolates of Fusarium graminearum were highly diverse in terms of vegetative compatibility grouping (VCG) and varied for production of ergosterol and mycotoxin production in rice culture. Aggressiveness was assessed by scoring the disease severity incited in wheat spikes by each isolate. Two inoculation methods, single-floret injection and spray of entire spikes, were used to screen 4 wheat varieties for reaction to the F. graminearum isolates. All isolates were of broadly similar aggressiveness, with disease severity ranging from 17.2 to 39.1 for single floret injection, and 39.1 to 69.0 for spray inoculation. Disease severity, ergosterol production, and mycotoxin development were not correlated. Using nitrate non-utilizing mutants the 15 isolates were grouped into 14 VCGs. Deoxynivalenol (DON) was produced by all isolates in rice culture, at levels between 0.2 and 249 ppm. 15-acetyldeoxynivalenol was produced by 14 of the 15 isolates at levels between 0.4 and 44.6 ppm. These results reveal a high level of diversity for several characteristics among F. graminearum isolates from Canada.

Trichothecene and Moniliformin Production by Fusarium Species from Western Canadian Wheat

Fusarium graminearum, Fusarium culmorum, and Fusarium avenaceum, isolated from Fusarium-damaged wheat harvested in western Canada, were cultured and evaluated for mycotoxin production. Extracts of the culture media were assayed for trichothecenes by gas chromatography-mass spectrometry and for moniliformin by liquid chromatography. Deoxynivalenol (DON) was found in 28 of 42 isolates of F. graminearum and 42 of 42 isolates of F. culmorum at levels ranging from 0.5 to 25.0 microg/g. 15-AcetylDON was found in 28 of 42 isolates of F. graminearum at levels ranging from 1.0 to 7.1 microg/g. 3-AcetylDON was found in 41 of 42 isolates of F. culmorum at levels ranging from 0.8 to 13.0 microg/g. Several other trichothecenes were assayed but not detected in the culture medium. Moniliformin was present in 40 of 42 isolates of F. avenaceum at levels ranging from 1.3 to 138.1 microg/g, but was not present in any of the isolates of F. graminearum or F. culmorum.

Mycotoxin formation in hulless barley during granary storage at 15 and 19% moisture content

Abstract Eleven-kilogram parcels of hulless barley ( Hordeum vulgare L. cv. Condor) at 15 and 19% initial moisture content were kept in simulated storage in a Manitoba farm granary for 20 weeks (June 1996–October 1996) to determine biotic and abiotic changes and mycotoxin production. Temperature, moisture content, CO 2 levels, ergosterol content, seed germination, microfloral infection, and the presence of major mycotoxins were monitored. Ochratoxin A, citrinin and sterigmatocystin reached mean levels of 24, 38 and 411 ppb by 20 weeks in the 19% moisture content barley, but were absent in the 15% moisture content barley; no other mycotoxins were detected. Penicillium species and Aspergillus versicolor (Vuill.) Tiraboschi comprised the predominant microflora. The effect of storage time was apparent at both 15 and 19% moisture content for grain temperature, Alternaria alternata (Fr.) Keissler, Penicillium species and Aspergillus versicolor . At 19% moisture content, storage time also affected moisture content, CO 2 level, ergosterol content, seed germination, and mycotoxin production. At 19% moisture content, elevated ergosterol levels at weeks 4 and 8 appear to offer early warning of the appearance of sterigmatocystin at week 12, and of ochratoxin A and citrinin at week 20.

Odor volatiles associated with microflora in damp ventilated and non-ventilated bin-stored bulk wheat

Western hard red spring wheat, stored at 20 and 25% moisture contents for 10 months during 1985-86, was monitored for biotic and abiotic variables in 10 unheated bins in Winnipeg, Manitoba. The major odor volatiles identified were 3-methyl-1-butanol, 3-octanone and 1-octen-3-ol. The production of these volatiles was associated and correlated with microfloral infection. Ventilation, used for cooling and drying of grain, disrupted microfloral growth patterns and production of volatiles. The highest levels of 3-methyl-1-butanol occurred in 25% moisture content wheat infected with bacteria, Penicillium spp. and Fusarium spp. In non-ventilated (control) bins with 20% moisture content wheat, 3-methyl-1-butanol was correlated with infection by members of the Aspergillus glaucus group and bacteria. In control bins, 1-octen-3-ol production was correlated with infection of wheat of both moisture contents by Penicillium spp. The fungal species, isolated from damp bin-stored wheat and tested for production of odor volatiles on wheat substrate, included Alternaria alternata (Fr.) Keissler, Aspergillus repens (Corda) Saccardo, A. flavus Link ex Fries, A. versicolor (Vuill.) Tiraboschi, Penicillium chrysogenum Thom, P. cyclopium Westling, Fusarium moniliforme Sheldon, F. semitectum (Cooke) Sacc. In the laboratory, fungus-inoculated wheat produced 3-methyl-1-butanol; 3-octanone and 1-octen-3-ol were also produced, but less frequently. Two unidentified bacterial species isolated from damp wheat and inoculated on agar produced 3-methyl-1-butanol.

Fungal volatiles associated with moldy grain in ventilated and non-ventilated bin-stored wheat.

The fungal odor compounds 3-methyl-l-butanol, l-octen-3-ol and 3-octanone were monitored in nine experimental bins in Winnipeg, Manitoba containing a hard red spring wheat during the autumn, winter and summer seasons of 1984–85. Quality changes were associated with seed-borne microflora and moisture content in both ventilated and non-ventilated bins containing wheat of 15.6 and 18.2% initial moisture content. All three odor compounds occurred in considerably greater amounts in bulk wheat in non-ventilated than in ventilated bins, particularly in those with wheat having 18.2% moisture content. The presence of these compounds usually coincided with infection of the seeds by the fungi Alternaria alternata (Fr.) Keissler, Aspergillus repens DeBarry, A. versicolor (Vuill.) Tiraboschi, Penicillium crustosum Thom, P. oxalicum Currie and Thom, P. aurantiogriseum Dierckx, and P. citrinum Thom. High production of all three odor compounds in damp wheat stored in non-ventilated bins was associated with heavy fungal infection of the seeds and reduction in seed germinability. High initial moisture content of the harvested grain accelerated the production of all three fungal volatiles in non-ventilated bins.

Immunochemical Detection of Molds: A Review

Molds are widely distributed in nature and cause deterioration of foods and feeds. Their mycotoxins can adversely affect human and animal health. Suitable assays for molds, therefore, are required to implement control and regulatory strategies and to develop appropriate feeding regimens for mold-infested feeds. Many different types of mold assays have been used, most of which are not reproducible or accurate. However, the immunoassays, particularly enzyme-linked immunosorbent assays (ELISAs), can be especially useful. Among these, assays that detect the water-soluble extracellular secretions of fungi, the exoantigens, are generally able to detect fungi at the genus or species level, whereas the heat-stable polysaccharides tend to be specific for one or more genus of fungi. Several species and genus (genera)-specific ELISAs have been developed using monoclonal or polyclonal antibodies against exoantigens and heat-stable polysaccharides from a wide range of fungi, including Aspergillus, Penicillium, and Fusarium species. Other assays have been developed that nonspecifically detect mold in food or feed, some using antibodies against a mixture of antigens from different fungi. These assays are highly sensitive, are easy to perform, and provide an index of the amount of mold present in the sample. Further refinement of these assays should facilitate their widespread use by food and feed processors, regulatory agencies, taxonomists, and research scientists.

Microbiological and aflatoxin evaluation of Brazil nut pods and the effects of unit processing operations.

Harvesting of Brazil nuts not only helps to preserve the Amazon rainforest but also provides income to individuals who would otherwise have little means of making a livelihood. Recently, the European Community has tightened the quality requirements for Brazil nuts, particularly with regard to aflatoxin levels and microbiological contamination. The objectives of this research were to gain a better understanding of the origin of aflatoxins on Brazil nuts and to microbiologically evaluate some of the operations involved in processing. In this regard, five Brazil nut pods were aseptically picked from trees located in each of three concessions of the Peruvian Amazon rainforest (Madre de Dios province). The exteriors of the pods and the nuts were examined for yeast and molds, including Aspergillus flavus and Aspergillus parasiticus, and for bacteria, including Salmonella and Escherichia coli. Brazil nuts obtained from various commercial process operations located in Peru were similarly evaluated. Exteriors of all Brazil nut pods did not contain A. parasiticus, and only pods from one concession yielded A. flavus isolates. All isolates tested were aflatoxigenic (630 to 915 ppb total aflatoxin). Coliforms, E. coli, and salmonellae were not recovered from any of the pods. Whole, in-shell nuts obtained after opening the pods yielded no A. flavus or A. parasiticus. Aflatoxins were not detected (detection limit 1.75 ppb) in any of the nuts. Whole, in-shell and shelled nuts from various process operations were all positive for A. flavus but negative for E. coli and salmonellae. Soaking of whole, in-shell nuts before cracking or shelling increased coliform numbers, whereas levels of A. flavus decreased. In order to gain a better understanding of the sanitary performance of the unit process operations, additional evaluations should be conducted on product lots processed on different days. Also, the microbiology of product processed from common lots should be followed through the various unit operations and compared.

Relationships among deoxynivalenol, ergosterol and Fusarium exoantigens in Canadian hard and soft wheat.

Soluble extracellular components (exoantigens) from cultures of Fusarium graminearum and F. sporotrichioides were used to produce antisera from chickens for an indirect enzyme immunoassay. This immunoassay was used to estimate Fusarium exoantigen levels in 40 samples of fusarium head blight-infected hard red spring wheat from Manitoba, and in 50 samples of infected soft white winter wheat from Ontario. These wheat samples were also assayed for deoxynivalenol (DON), the predominant Fusarium mycotoxin, and for ergosterol, a metabolite reflecting fungal biomass. Using F. sporotrichioides antisera, the linear correlations between exoantigen level and DON content for the hard and soft wheats had coefficients of 0.80 and 0.76, respectively. With the same antisera, linear correlations between exoantigen level and total ergosterol concentration for the hard and soft wheats had coefficients of 0.66 and 0.81, respectively.

The incidence and distribution of ochratoxin A in western Canadian swine

A survey of swine destined for slaughter in Manitoba was conducted to examine the incidence of ochratoxin A (OA) in swine herds from different regions of Manitoba throughout the year 1989-90. Thirty-six percent of the serum samples which were collected from 1600 pigs contained detectable levels of OA. The identity of this toxin was confirmed using liquid chromatography-mass spectrometry and enzymatic hydrolysis. There was a significant effect of the region from which the herds originated, as well as the season in which the samples were collected on both the incidence (p < 0.001) and concentration of OA (p < 0.001). In July, 65% of the samples contained detectable levels of OA, compared with 38, 21 and 17%, in April, October and January respectively. Furthermore, 24% of the samples collected in July contained greater than 15 ng/ml of OA, while only 2, 9, and 1% of the samples collected in April, October and January respectively, contained greater than 15 ng/ml of OA. Based on the six samples collected from each herd, it appears that the presence and concentration of OA within a herd may be estimated from a limited number of animals per herd.

NephrotoxigenicPenicillium species occurring on farm-stored cereal grains in western Canada

The incidence of nephrotoxigenicPenicillium species on farm-stored cereals in western Canada was determined by morphological and metabolite profile examination. Of the 142 isolates examined 102 were toxin producers with 61P. aurantiogriseum and 27P. freii. Other nephrotoxigenic species includedP. tricolor (6 isolates),P. verrucosum Chemotype II (4 isolates) andP. viridicatum Westling (4 isolates). The nephrotoxigenicPenicillium species profile for western Canada appears to differ from that of Denmark whereP. verrucosum,P. cyclopium,P. freii and, to a lesser extent,P. aurantiogriseum,P. polonicum, andP. viridicatum predominate.