Isolation of culturable mycobiota from agricultural soils and determination of tolerance to glyphosate of nontoxigenic Aspergillus section Flavi strains.

Glyphosate-based herbicides are extensively used in Argentina's agricultural system to control undesirable weeds. This study was conducted to evaluate the culturable mycobiota [colony forming units (CFU) g(-1) and frequency of fungal genera or species] from an agricultural field exposed to pesticides. In addition, we evaluated the tolerance of A. oryzae and nontoxigenic A. flavus strains to high concentrations (100 to 500 mM - 17,000 to 84,500 ppm) of a glyphosate commercial formulation. The analysis of the mycobiota showed that the frequency of the main fungal genera varied according to the analyzed sampling period. Aspergillus spp. or Aspergillus section Flavi strains were isolated from 20 to 100% of the soil samples. Sterilia spp. were also observed throughout the sampling (50 to 100%). Aspergillus section Flavi tolerance assays showed that all of the tested strains were able to develop at the highest glyphosate concentration tested regardless of the water availability conditions. In general, significant reductions in growth rates were observed with increasing concentrations of the herbicide. However, a complete inhibition of fungal growth was not observed with the concentrations assayed. This study contributes to the knowledge of culturable mycobiota from agricultural soils exposed to pesticides and provides evidence on the effective growth ability of A. oryzae and nontoxigenic A. flavus strains exposed to high glyphosate concentrations in vitro.

Potentiation of the effect of a commercial animal feed additive mixed with different probiotic yeast strains on the adsorption of aflatoxin B1.

This study potentiates the adsorbent effect for aflatoxin B1 (AFB1) of a commercial additive (CA) of animal feed, containing inactive lysate of three Saccharomyces cerevisiae strains, active enzymes, adsorbents and a selenium-amino acid complex, when the additive was mixed separately with three S. cerevisiae strains. Levels of AFB1 of 20 and 50 ng g(-1) were used to determine the binding capacity of different concentrations of CA alone and in the presence of yeast strains, as well as toxin desorption, under gastrointestinal conditions. The viability of yeasts in the presence of CA was evaluated. The results show that the CA did not affect the viability of the yeast strains assayed. CA alone showed a low percentage adsorption. At 20 and at 50 ng g(-1), CA was highly efficient in adsorbing AFB1 when combined with RC016 and RC012 strains respectively. Desorption of AFB1 by CA alone and in combination with the yeasts increased with increasing levels of CA. The results demonstrate the improvement of CA in AFB1 adsorption once it is mixed with live yeasts.

Inhibitory effect of sodium metabisulphite and chlorine on growth of Aspergillus spp. and Penicillium spp. strains isolated from marine shrimp / Efeito inibitório do metabissulfito de sódio e do cloro sobre o crescimento de cepas de Aspergillus spp. e Penicilium spp. isoladas de camarão marinho

The sodium metabisulphite (SMB) is used in shrimp farming to prevent melanosis and the 5.0 ppm chlorine (CL) concentration used in the shrimp processing is efficient as a bactericide, but there is no evidence of the effectiveness of these chemical compounds as fungicides. Therefore, the aim of this study was to evaluate the in vitro effect of sodium metabisulphite (SMB) and chlorine (CL) on the growth of Aspergillus and Penicillium species isolated from marine shrimp in different stages of processing. The samples were collected from a frozen shrimp processing industry, located in Piauí State, Brazil. The total fungi and occurrence of Aspergillus and Penicillium species were evaluated. For in vitro sensibility test using the diffusion disk in agar method, five concentrations of SMB (0%, 1%, 3%, 5% and 10%) and six of CL (0, 1, 2, 3, 4 and 5 µg mL-1) were used. The fungal counts in the different processing stages ranged from 1.74 to 3.38 CFU g-1. Twenty-nine Aspergillus strains were isolated, prevailing A. versicolor (59.3%) and twenty-two of Penicillium, prevailing P. citrinum (74%). One strain of A. flavus was AFB1 producer. All the isolated strains of P. citrinum produced citrinin. All tested species were in vitro sensitive to 3% of SMB, except the A. flavus. The 10% concentration of SMB inhibited the in vitro growth of all strains. The CL concentrations tested did not inhibit the studied species growth and SMB concentrations above 3.0% inhibited in vitro the growth of the tested strains.

O metabissulfito de sódio (SMB) é usado na carcinicultura para evitar a melanose e a concentração de 5,0ppm de cloro (CL), utilizada no beneficiamento do camarão, é eficiente como bactericida, porém não há comprovação da eficácia destes compostos químicos como fungicida. Desse modo, o objetivo deste estudo foi avaliar o efeito in vitro do metabissulfito de sódio (SMB) e cloro (CL) sobre o crescimento de espécies de Aspergillus e Penicillium isolados de camarão marinho em diferentes estágios de processamento. As amostras foram coletadas de uma indústria de processamento de camarão congelado, localizada no Estado do Piauí, Brasil. Fungos totais e ocorrência das espécies de Aspergillus e Penicillium foram avaliados. Para o teste in vitro de sensibilidade pelo método disco-difusão em ágar, foram utilizadas cinco concentrações de SMB (0%, 1%, 3%, 5% e 10%) e seis de CL (0, 1, 2, 3, 4 e 5µg mL-1). As contagens fúngicas nos diferentes estágios de processamento variaram de 1,74 a 3,38UFC g-1. Foram isoladas 29 cepas de Aspergillus, prevalecendo o A. versicolor (59,3%) e 22 de Penicillium, prevalecendo o P. citrinum (74%). Uma cepa de A. flavus era produtora de AFB1. Todas as cepas de P. citrinum isoladas produziram citrinina. Todas as espécies testadas foram sensíveis in vitro a 3% do SMB, com exceção do A. flavus. A concentração de 10% do SMB inibiu in vitro o crescimento de todas as cepas. As concentrações de CL testadas não inibiram o crescimento das espécies isoladas e concentrações de SMB acima de 3,0% inibiram in vitro o crescimento das linhagens testadas.

Effect of acid lactic bacteria isolated from faeces of healthy dogs on growth parameters and aflatoxin B1 production by Aspergillus species in vitro.

The aim of the present study was to evaluate the inhibitory effect of Enterococcus faecium and Lactococcus lactis subsp. lactis isolated from faeces of healthy dogs on (i) lag phase, (ii) growth rate, and (iii) aflatoxin B1 production by Aspergillus section Flavi on in vitro assays. Thirteen lactic acid bacteria (LAB) isolates were used as antagonist microorganisms. Antagonistic activity was assayed against four potentially aflatoxigenic Aspergillus section Flavi isolates: A. flavus (AF210 and AF281), A. parasiticus (AP245) and A. parasiticus (NRRL 2999). In general, the longest lag phases of Aspergillus isolates were obtained with E. faecium GJ40. Respecting the growth rate, no significant reduction was found in this parameter in the interaction assays with A. flavus and antagonist isolates respecting the control. While in A. parasiticus a significant reduction in growth rate was only observed in the interaction among reference strain and E. faecium MF5 isolate (p < 0.05). In general, AFB1 production was reduced by most of the LAB isolates assayed, except for E. faecium GJ18, GJ20, MF3 and MF4. This study provides the first data about the antiaflatoxigenic activity of autochthonous LAB isolated from dog faeces.

Fungi and mycotoxins in feed intended for sows at different reproductive stages in Argentina.

The aim of this study was to evaluate fungi and contamination levels of aflatoxin B(1), ochratoxin A, fumonisin B(1), and zearalenone in raw materials and finished feed intended for sows at different reproductive stages. Total fungi, Aspergillus, Penicillium, and Fusarium species occurrence, were examined. Aspergillus flavus, A. niger aggregate spp., and F. verticillioides were the prevalent species. Fungal counts exceeded the levels proposed as feed hygienic quality limits (1 x 10(4) colony forming units) at all reproductive stages. Aflatoxin B(1), ochratoxin A, fumonisin B(1), and zearalenone were detected by high-pressure liquid chromatography. Aflatoxin levels in 80% samples of finished sow feeds were over the permitted levels of 0.02 mug g(-1) (mean 228.2 +/- 95 mug Kg(-1)). Fumonisin B(1) was detected in all tested raw materials at levels that varied from 50.3 to 1137.64 mug Kg(-1) and finished feed samples at levels that ranged from 99.8 to 512.4 mug Kg(-1). Aflatoxin B(1), zearalenone, and ochratoxin A were not detected in raw materials. All finished feeds were negative for zearalenone contamination whereas all nonpregnant gilt samples were contaminated with low OTA levels (mean 0.259 +/- 0.123). This fact requires periodic monitoring to prevent the occurrence of mycotoxicosis in animal production, to reduce the economic losses, and to minimize hazards to human health.

Effect of antioxidant mixtures on growth and ochratoxin a production of Aspergillus section Nigri species under different water activity conditions on peanut meal extract agar.

The effect of mixtures of antioxidants butylated hydroxyanisol (BHA) and propyl paraben (PP) on lag phase, growth rate and ochratoxin A (OTA) production by four Aspergillus section Nigri strains was evaluated on peanut meal extract agar (PMEA) under different water activities (a(w)). The antioxidant mixtures used were: BHA + PP (mM), M1 (0.5 + 0.5), M2 (1.0 + 0.5), M3 (2.5 + 0.5), M4 (0.5 + 1.0), M5 (1.0 + 1.0), M6 (2.5 + 1.0), M7 (5.0 + 2.5) and M8 (10 + 2.5). The mixture M8 completely suppressed mycelial growth for all strains. A significant stimulation in OTA production was observed with mixtures M1 to M5 mainly at the highest a(w); whereas M6, M7 and M8 completely inhibited OTA production in all strains assayed; except M6 in A. carbonarius strain (RCP G). These results could enable a future intervention strategy to minimize OTA contamination.

Ochratoxin A and Aspergillus section Nigri in peanut seeds at different months of storage in Córdoba, Argentina.

Peanut is an important food commodity in Argentina. Last year Córdoba Province accounted for approximately 96% of the total Argentinian production. Few surveys of peanuts for the natural occurrence of ochratoxins and ochratoxin-producing fungi have been reported. The objectives of this study were to investigate the occurrence of Aspergillus section Nigri and ochratoxin A (OTA) in storage peanuts during a three-month-period. The capacity to produce OTA by Aspergillus section Nigri was also studied. A total of 100 samples were collected from May to July 2004. The frequency of contaminating fungi were determined by surface-disinfection the seeds and plating onto several agar types. Detection of OTA in seed samples was performed using an HPLC method. Strains belonging to Aspergillus section Nigri or Flavi were detected in all seeds samples. From the section Nigri, the species belonging to A. niger aggregate were isolated in 100% of the samples. The main ochratoxigenic specie, A. carbonarius, was present at low levels throughout the study period. OTA was found in 50% of the peanut samples, with mean levels ranging from 5.6 to 130 ng g(-1). The mean value of OTA obtained after the first month of storage (30 ng g(-1)) was significantly higher from those obtained after the second (6.5 ng g(-1)) and third (13 ng g(-1)) month (p<0.0001). One hundred and four (32%) of 322 strains of Aspergillus section Nigri, were OTA producers. The levels of toxin produced ranged from 2 to 24 ng ml(-1) of culture medium (mean level: 12.7 ng ml(-1)). These results indicate that humans and animals being may be frequently exposed to OTA in Argentina through the ingestion of peanut seed and foods based on peanuts. The presence of this toxin in peanuts might be an appropriate focus for future studies to estimate exposure through normal consumption of this commodity. These data are important in formulating guidelines for quality control of peanuts in Argentina.