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Background  Brazil nut is a protein-rich extractivist tree crop in the Amazon region. Fungal contamination of shells and kernel material frequently includes the presence of aflatoxigenic  Aspergillus  species from the section  Flavi . Aflatoxins are polyketide secondary metabolites, which are hepatotoxic carcinogens in mammals. The objectives of this study were to identify  Aspergillus  species occurring on Brazil nut grown in different states in the Brazilian Amazon region and develop a specific PCR method for collective identification of member species of the genus  Aspergillus.Results  Polyphasic identification of 137  Aspergillus  strains isolated from Brazil nut shell material from cooperatives across the Brazilian Amazon states of Acre, Amapá and Amazonas revealed five species, with  Aspergillus  section  Flavi  species  A. nomius  and  A. flavus  the most abundant. PCR primers ASP_GEN_MTSSU_F1 and ASP_GEN_MTSSU_R1 were designed for the genus  Aspergillus , targeting a portion of the mitochondrial small subunit ribosomal RNA gene. Primer specificity was validated through both electronic PCR against target gene sequences at Genbank and in PCR reactions against DNA from  Aspergillus  species and other fungal genera common on Brazil nut. Collective differentiation of the observed section  Flavi  species  A. flavus ,  A. nomius  and  A. tamarii  from other  Aspergillus  species was possible on the basis of RFLP polymorphism.    Conclusions  Given the abundance of  Aspergillus  section  Flavi  species  A. nomius  and  A. flavus  observed on Brazil nut, and associated risk of mycotoxin accumulation, simple identification methods for such mycotoxigenic species are of importance for Hazard Analysis Critical Control Point system implementation. The assay for the genus  Aspergillus  represents progress towards specific PCR identification and detection of mycotoxigenic species.

bmc microbiology

Characterization of Aspergillus species on Brazil nut from the Brazilian Amazonian region and development of a PCR assay for identification at the genus level