Graciela Vaamonde

Variability of aflatoxin and cyclopiazonic acid production by Aspergillus section flavi from different substrates in Argentina.

Aspergillus section flavi strains isolated from peanuts, wheat and soybean grown in Argentina were screened for aflatoxins (type B and G) and cyclopiazonic acid (CPA) production. Aspergillus flavus was the predominant species in all substrates, although there was almost the same proportion of A. flavus and Aspergillus parasiticus in peanuts. Aspergillus nomius was not found. Incidence of aflatoxigenic A. flavus strains was higher in peanuts (69%) than in wheat (13%) or soybeans (5%) while the ratio of CPA producers A. flavus isolated from all substrates was very high (94% in peanuts, 93% in wheat and 73% in soybeans). Isolates of A. flavus able to produce simultaneously aflatoxins type B and CPA were detected in all substrates, suggesting the possibility of co-occurrence of these toxins. Almost all isolates of A. parasiticus resulted aflatoxins (type B and G) producers but did not produce CPA. Five of sixty-seven strains isolated from peanuts showed an unusual pattern of mycotoxin production (aflatoxins type B and G simultaneously with CPA). These strains also produced numerous small sclerotia like S strains of A. flavus detected in cottonseed in Arizona and in soils of Thailand and West Africa. The atypical strains are not widely distributed in Argentina and were found uniquely in peanuts.

Natural co-occurrence of aflatoxin and cyclopiazonic acid in peanuts grown in Argentina

Natural occurrence of aflatoxins and cyclopiazonic acid (CPA) contamination in peanuts was investigated. Co-occurrence of CPA and aflatoxins was detected in two of 50 samples analysed. The levels of these toxins found in positive samples were 4300 and 493 μg kg -1 for CPA, 625 and 435 μg kg -1 for aflatoxin B1 (AFB1), and 625 and 83 μg kg -1 for aflatoxin G1 (AFG1), respectively. Levels of CPA contamination in the positive samples were similar to those registered in other substrates. This is the first report of natural co-occurrence of CPA and aflatoxins in Argentina.

Influence of water activity on Penicillium citrinum growth and kinetics of citrinin accumulation in wheat

The influence of water activity (aw) on both Penicillium citrinum growth and citrinin accumulation in wheat was studied. Wheat conditioned at different levels of aw and inoculated with a citrinin producer strain was incubated at 30 degrees C for 2 months. Fungal growth was assessed by microscopic examination. P. citrinum grew down to aw 0.775. Citrinin was not detected in the substrate at aw 0.800 and lower. As aw increased the toxin was detected earlier and the maximum accumulation increased markedly (65 microg/kg at aw 0.810, 460 microg/kg at aw 0.825 and 22 mg/kg at aw 0.885). Citrinin concentration declines rapidly after reaching the maximum at each aw level.

Modelling the effect of temperature and water activity of Aspergillus flavus isolates from corn

The aim of this study was to model the effects of temperature (10-40 °C) and a(w) (0.80-0.98), in two media (Czapek yeast agar: CYA; corn extract medium: CEM) on the growth rates and growth boundaries (growth-no growth interface) of three strains of A. flavus isolated from corn in Argentina. Both kinetic and probability models were applied to colony growth data. The growth rates obtained in CYA were significantly (p<0.05) greater than those obtained in CEM medium. No significant differences (p<0.05) were observed among the three isolates. The growth rate data showed a good fit to the Rosso cardinal models combined with the gamma-concept with R(2)=0.98-0.99 and RMSE=0.60-0.78, depending on media and isolates. The probability model allowed prediction of safe storage (p of growth <0.01) for one month for moist maize (e.g. 0.90 a(w)) provided temperature is under 15 °C, or for dry maize (e.g. 0.80 a(w)) provided temperature is under 27 °C. Storage at <0.77 a(w) would be safe regardless of the storage temperature. Probability models allow evaluation of the risk of fungal contamination in the process of storage, so the results obtained in this study may be useful for application in systems of food safety management.

A survey on distribution and toxigenicity of Aspergillus section Flavi in poultry feeds

Thirty-five samples of poultry feeds and corresponding raw materials (maize, soybean and meat meal) from a processing plant were analyzed to evaluate the distribution and toxigenicity of Aspergillus section Flavi isolates. Mycological analysis of the samples indicated the presence of five fungal genera (Aspergillus, Penicillium, Fusarium, Cladosporium, and Eurotium). Aspergillus flavus was the predominant species being present in 48.5% of the analyzed samples. Ninety-one isolates belonging to Aspergillus section Flavi were isolated; ninety were identified as A. flavus and only one as A. parasiticus. Fifty-seven isolates were capable of producing sclerotia, 41 were identified as L-type strains and 16 as type S. Fifty-seven percent of the isolates produced AFB₁ levels ranging from 0.05 μg/kg to 27.7 μg/kg whereas 86.8% produced CPA from 1.5 μg/kg to 137.8 μg/kg. L-strains produced from 0.05 to 14.8 μg/kg of aflatoxin and type S produced levels from 0.05 to 1.65 μg/kg. No significant differences in CPA production among S- and L-strains were observed. Sclerotial isolates produced AFB₁ levels ranging between 0.05 and 27.7 μg/kg and CPA levels from 3.8 to 47.3 μg/kg. More than half of the A. flavus isolates were able to produce AFB and CPA simultaneously. Twenty percent of the 35 samples were contaminated with aflatoxin B₁ whereas 34.3% were contaminated with CPA. The high rate of CPA producing isolates represents a potential risk of contamination with this toxin in poultry feeds.

Ochratoxin A production by a mixed inoculum of Aspergillus carbonarius at different conditions of water activity and temperature.

Growth rate, lag phase and OTA production of a mixed inoculum of four Aspergillus carbonarius strains were compared to the behaviour of the single strains at 30 degrees C on Czapek Yeast Extract Agar. Significant differences between radial growth rates and lag phases of the different isolates were observed; however, no significant differences between growth rates of each strain and the mixed inoculum were detected. When the four strains were inoculated simultaneously, the lag phase was the same than the higher value obtained for individual strains, suggesting mycelial interactions between the A. carbonarius isolates. The four strains differed in maximum OTA yield, and the toxin accumulation by the mixed inoculum showed intermediate levels at each time point. The effects of water activity (0.83, 0.85, 0.87, 0.89, 0.90, 0.93 and 0.95) and temperature (15, 20, 25, 30, and 35 degrees C) on OTA production by the mixed inoculum were studied at 7, 14, 21 and 28days of incubation. The limiting water activity for OTA production by the mixed inoculum was 0.87, showing xerotolerant behaviour of the strains isolated from dried vine fruits. Results obtained were similar to those reported for single A. carbonarius strains from European countries, Israel, Australia and South America. The similar trend in the response of the different isolates to the variation of environmental parameters may be of interest for the building of predictive models.

Influence of inoculum size on growth rate and lag phase of fungi isolated from Argentine corn

Abstract The influence of the inoculum size on growth of nine strains of fungi belonging to the genera Aspergillus, Penicillium and Fusarium was investigated. The lag phase and the growth rate were evaluated by using a synthetic medium. The growth rate was not significantly affected by the initial number of spores varying from 10 to 10 5 . The lag phase increase with decreasing number of spores, and linear correlation between the log of the inoculum size and lag phase was observed for Aspergillus niger, A. flavus, A. wentii, A. parasiticus, Penicillium citrinum, Fusarium moniliforme and F. semitectum . A different behavior was found for A. repens and P. chrysogenum .

Mycotoxin-producing potential of fungi isolated from amaranth seeds in Argentina.

To evaluate the potential for mycotoxin production by molds in amaranth grains, the mycoflora was determined both before and after surface disinfection on dichloran-chloramphenicol-peptone agar (DCPA) and dichloran-18% glycerol agar (DG18). On both media Aspergillus, Penicillium, Fusarium and Alternaria were the predominant genera. A flaus and A. parasiticus were the Aspergillus species most frequently isolated. P. chrysogenum was the species most common among the penicillia. F. equiseti was the predominant Fusarium species. Isolates of Aspergillus, Penicillium and Fusarium were screened for mycotoxin production on sterile rice substrate and also using a simple screening method. Toxinogenic strains of A. flavus and A. parasiticus (aflatoxins), A. versicolor (sterigmatocystin), P. citrinum (citrinin), P. viridicatum (penicillic acid), F. moniliforme, F. equiseti and F. semitectum (zearalenone), were encountered. The simple screening method for toxinogenic molds showed good performance for the detection of molds producing aflatoxins and zearalenone compared with mycotoxins production on the natural substrate.

Amaranth grain as substrate for aflatoxin and zearalenone production at different water activity levels

The influence of water activity (a(w)) on the kinetics of aflatoxin and zearalenone production in amaranth grains at 25 degrees C was studied. Minimum a(w) for aflatoxin production in this substrate was 0.825. Accumulation of the four aflatoxins (B1, B2, G1 and G2) was similar at a(w) 0.825 (maximum 81.2 microg/kg after 42 days) and 0.868 (maximum 109.6 microg/kg after 49 days). Maximum accumulation of total aflatoxins at a(w) 0.902 (260.4 microg/kg) was detected after 21 days, with an appreciable increment in the concentration of aflatoxins B1 and G1. These quantities were lower than those reported for aflatoxin production on other cereals and legumes, indicating that amaranth is not a good substrate for aflatoxin production. Zearalenone was not detected at a(w) 0.902. Maximum accumulation of zearalenone was 1.5 microg/g after 35 days at a(w) 0.925 and 11.1 microg/g after 49 days at a(w) 0.950.

Influence of water activity and temperature on the accumulation of zearalenone in corn

The influence of water activity (aw) and temperature on the zearalenone biosynthesis in corn has been examined. Viable corn kernels were conditioned at different values of water activity (0.90, 0.95 and 0.97), inoculated with Fusarium graminearum and incubated at different temperatures. Zearalenone was determined at selected times. For the strain used, a constant temperature of 25 degrees C resulted more favorable than 15 degrees C, 20 degrees C and the combination of two weeks at 25 degrees C followed by incubation at 15 degrees C shown by other authors to increase the zearalenone accumulation. Both F. graminearum growth and zearalenone production at 25 degrees C were inhibited at water activity 0.90. At short incubation times, toxin accumulation was greater at water activity 0.97 than at water activity 0.95 (25 degrees C). This relation was inverted at longer periods of incubation. A combined effect of water activity and temperature on the zearalenone accumulation was observed.