Patricia Zorzete

Fungal diversity and natural occurrence of deoxynivalenol and zearalenone in freshly harvested wheat grains from Brazil

This study investigated the fungal diversity and presence of deoxynivalenol and zearalenone in 150 samples of freshly harvested wheat grains collected in three regions of Brazil (Sao Paulo, Parana, and Rio Grande do Sul). Analysis of the mycobiota showed a predominance of Alternaria sp., Fusarium sp. and Epicoccum sp. Microdochium nivale (23%), a fungus rarely found in Brazilian crops, was detected in Sao Paulo. Four members of the Fusarium graminearum species complex were isolated: F. graminearum s.s. (37%), Fusarium meridionale (46%), Fusarium cortaderiae (13%), and Fusarium austroamericanum (3%). Toxin analysis revealed 99% contamination with deoxynivalenol (mean 706 μg/kg). The frequency of zearalenone varied greatly across regions: wheat grains from Rio Grande do Sul (84%) and Sao Paulo (12%) had median concentrations of 70.9 and 57.9 μg/kg, respectively. ZEA was not detected in the samples from Parana. A total of six samples were above the maximum tolerated level recommended by the European Commission for ZEA in wheat grains. This study provided new insights into the natural mycobiota of Brazilian wheat, demonstrating contamination of most samples with deoxynivalenol and high frequency of zearalenone in samples from Rio Grande do Sul.

Mycobiota and mycotoxins in Brazil nut samples from different states of the Brazilian Amazon region

The objective of this study was to evaluate the presence of fungi and mycotoxins (aflatoxins and cyclopiazonic acid) in Brazil nut samples collected in different states of the Brazilian Amazon region: Acre, Amazonas, Amapá, and Pará. A total of 200 husk samples and 200 almond samples were inoculated onto Aspergillus flavus-parasiticus agar for the detection of fungi. Mycotoxins were analyzed by high-performance liquid chromatography. The mycobiota comprised the following fungi, in decreasing order of frequency: almonds - Phialemonium spp. (54%), Penicillium spp. (16%), Fusarium spp. (13%), Phaeoacremonium spp. (11%), and Aspergillus spp. (4%), husks - Phialemonium spp. (62%), Phaeoacremonium spp. (11%), Penicillium spp. (10%), Fusarium spp. (9%), and Aspergillus spp. A polyphasic approach was used for identification of Aspergillus species. Aflatoxins were detected in 22 (11%) of the 200 almond samples, with 21 samples presenting aflatoxin B(1) levels above 8μg/kg, the limit established by the European Commission for Brazil nuts for further processing. Nineteen (9.5%) of the 200 husk samples contained aflatoxins, but at levels lower than those seen in almonds. Cyclopiazonic acid (CPA) was detected in 44 (22%) almond samples, with levels ranging from 98.65 to 161.2μg/kg. Aspergillus nomius and A. flavus were the most frequent Aspergillus species. The presence of fungi does not necessarily imply mycotoxin contamination, but almonds of the Brazil nut seem to be a good substrate for fungal growth.

Polyphasic approach to the identification of Aspergillus section Flavi isolated from Brazil nuts.

The aim of this study was to use a polyphasic approach to identify Aspergillus section Flavi isolated from Brazil nuts collected in the Amazon forest: investigation of macro- and microscopic morphology, production of extrolites, heat-resistance fungi, and sequencing of DNA regions. The following Aspergillus section Flavi species were identified: Aspergillus flavus (75.5%), Aspergillus nomius (22.3%), and Aspergillus parasiticus (2.2%). All A. nomius and A. parasiticus isolates produced aflatoxins B and G, but not cyclopiazonic acid (CPA). A. flavus isolates were more diversified and a high frequency of mycotoxigenic strains was observed. The polyphasic approach permitted the reliable identification of section Flavi species. The rate of mycotoxigenic strains was high (92.7%) and mainly included A. flavus strains producing elevated levels of aflatoxins and CPA. These results highlight the possibility of co-occurrence of both toxins, increasing their potential toxic effect in this commodity.

Fungi, mycotoxins and phytoalexin in peanut varieties, during plant growth in the field

The aim of the present study was to analyze the mycobiota, occurrence of mycotoxins (aflatoxins and cyclopiazonic acid), and production of phytoalexin (trans-resveratrol) in two peanut varieties (Runner IAC 886 and Caiapó) during plant growth in the field. Climatic factors (rainfall, relative humidity and temperature) and water activity were also evaluated. The results showed a predominance of Fusarium spp. in kernels and pods, followed by Penicillium spp. and Aspergillus flavus. Aflatoxins were detected in 20% and 10% of samples of the IAC 886 and Caiapó varieties, respectively. Analysis showed that 65% of kernel samples of the IAC 886 variety and 25% of the Caiapó variety were contaminated with cyclopiazonic acid. trans-Resveratrol was detected in 6.7% of kernel samples of the IAC 886 variety and in 20% of the Caiapó variety. However, trans-resveratrol was found in 73.3% of leaf samples in the two varieties studied.

Mycoflora and fumonisin contamination in Brazilian sorghum from sowing to harvest

BACKGROUND The aim of this study was to characterise the mycoflora and the presence of fumonisin in sorghum grains, correlating the results with the environment and abiotic factors. RESULTS Fifty samples (five collections of ten samples each) of sorghum were analysed. All samples were found to be contaminated with fungi, with higher frequencies of Cladosporium spp. (61.8%) and Helminthosporium spp. (33.4%). Fusarium verticillioides was isolated from 15.1% of the samples, with 38% of them being contaminated with fumonisin B(1) (FB(1)) at levels ranging from 50 to 368.78 ng g(-1). Regarding abiotic factors, temperature, water activity and rainfall showed a positive correlation with the frequency of F. verticillioides and FB(1) production. There was a significant positive correlation between relative air humidity and FB(1) production. The results obtained from sexual crosses between standard F mating tester strains and the isolated strains confirmed that the strains isolated were F. verticillioides. CONCLUSION It can be concluded that the decrease in F. verticillioides and fumonisin contamination occurred owing to atypical climatic factors during the period of sorghum cultivation, when there was any occurrence of rain and the level of water activity of grains did not reach 0.58.

Monitoring and Determination of Fungi and Mycotoxins in Stored Brazil Nuts

Brazil nut (Bertholletia excelsa) is an important commodity from the Brazilian Amazon, and approximately 37,000 tons (3.36 × 10⁷ kg) of Brazil nuts are harvested each year. However, substantial nut contamination by Aspergillus section Flavi occurs, with subsequent production of mycotoxins. In this context, the objective of the present investigation was to evaluate the presence of fungi and mycotoxins (aflatoxins and cyclopiazonic acid) in 110 stored samples of cultivated Brazil nut (55 samples of nuts and 55 samples of shells) collected monthly for 11 months in Itacoatiara, State of Amazonas, Brazil. The samples were inoculated in duplicate onto Aspergillus flavus and Aspergillus parasiticus agar and potato dextrose agar for the detection of fungi, and the presence of mycotoxins was determined by high-performance liquid chromatography. The most prevalent fungi in nuts and shells were Aspergillus spp., Fusarium spp., and Penicillium spp. A polyphasic approach was used for identification of Aspergillus species. Aflatoxins and cyclopiazonic acid were not detected in any of the samples analyzed. The low water activity of the substrate was a determinant factor for the presence of fungi and the absence of aflatoxin in Brazil nut samples. The high frequency of isolation of aflatoxigenic Aspergillus section Flavi strains, mainly A. flavus, and their persistence during storage increase the chances of aflatoxin production on these substrates and indicates the need for good management practices to prevent mycotoxin contamination in Brazil nuts.