Título:	Characterization of Aspergillus species on Brazil nut from the Brazilian Amazonian region and development of a PCR assay for identification at the genus level
Autor(es):	Midorikawa, Gláucia Emy Okida Sousa, Maria de Lourdes M. de Silva, Otniel Freitas Dias, Jurema do Socorro A. Kanzaki, Luis Isamu Barros Hanada, Rogerio E. Mesquita, Renata M. L. C. Gonçalves, Rivadalve C. Alvares, Virginia S. Bittencourt, Daniela Matias de Carvalho Miller, Robert Neil Gerard
Assunto:	Castanha-do-Pará Amazônia Fungos
Data de publicação:	30-Mai-2014
Editora:	BioMed Central
Referência:	MIDORIKAWA, Gláucia Emy Okida et al. Characterization of Aspergillus species on Brazil nut from the Brazilian Amazonian region and development of a PCR assay for identification at the genus level. BMC Microbiology, v. 14, Article 138, 30 mai. 2014. Disponível em: <https://bmcmicrobiol.biomedcentral.com/articles/10.1186/1471-2180-14-138>. Acesso em: 19 jun. 2017. doi: https://bmcmicrobiol.biomedcentral.com/articles/10.1186/1471-2180-14-138.
Abstract:	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.
Licença:	© 2014 Midorikawa et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
DOI:	https://dx.doi.org/10.1186/1471-2180-14-138
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