Bioprospecting Aspergillus section Nigri in Atlantic Forest soil and plant litter / Bioprospecção de Aspergillus seção Nigri em solo e serrapilheira da Mata Atlântica

The use of fungi as a source of enzymes has become widespread in various industrial and commercial areas, and Aspergillus section Nigri has significant potential for producing enzymes. The aim of this study was to isolate Aspergillus section Nigri from plant litter and soil from the Atlantic Forest biome and evaluate it with regards to hydrolytic enzyme production. The trials for producing the enzymes were carried out in Petri dishes, using different culture mediums adapted for microbial growth and with the respective substrates for inducing enzyme production - cellulase (carboxymethyl cellulose), protease (skimmed milk), amylase (soluble starch), pectinase (citrus pectin), and phytase (Pikovskaya medium). Forty-two fungi were isolated, 16.7% derived from the plant litter layer and 83.3% derived from soil at a depth of 0 to 5 cm and 5 to 10 cm. All of the isolated lineages presented amylase, protease, and phytase production, with 90.4% producing cellulase and no lineage producing pectinase. From the results, the significant potential for Atlantic Forest fungi as hydrolytic enzyme producers could be perceived. The enzymatic activity evaluations presented a satisfactory result when compared with the scientific literature.(AU)

A utilização dos fungos como fonte de enzimas vem adquirindo status de destaque nas mais variadas áreas industriais e comerciais, e os Aspergillus membros da seção Nigri possuem significativo potencial para produção de enzimas. Os objetivos deste estudo foram isolar e avaliar Aspergillus da seção Nigri de serrapilheira e solos do bioma Mata Atlântica quanto à produção de enzimas hidrolíticas. Os ensaios para produção das enzimas foram realizados em placas de Petri, utilizando diferentes meios de cultivo, adequados ao crescimento microbiano e com a presença dos respectivos substratos indutores à produção das enzimas - celulases (carboximetilcelulose), proteases (leite desnatado), amilases (amido solúvel), pectinases (pectina cítrica) e fitase (meio Pikovskaya). Foram isolados 42 fungos, sendo desse total 16,7% provenientes da camada de serrapilheira e 83,3% provenientes do solo na profundidade de 0 a 5 cm e 5 a 10 cm. Todas as linhagens isoladas apresentaram produção de amilases, protease e fitase, 90,4% produziram celulase, e nenhuma linhagem produziu pectinase. Com esses resultados, percebeu-se significativo potencial dos fungos da Mata Atlântica como produtores de enzimas hidrolíticas. As avaliações da atividade enzimática apresentaram resultado satisfatório quando comparados à literatura científica.(AU)

Classification and infection mechanism of entomopathogenic fungi / Classificacão e mecanismo de infecção dos fungos entomopatogênicos

Entomopathogenic fungi are important biological control agents throughout the world, have been the subject of intensive research for more than 100 years, and can occur at epizootic or enzootic levels in their host populations. Their mode of action against insects involves attaching a spore to the insect cuticle, followed by germination, penetration of the cuticle, and dissemination inside the insect. Strains of entomopathogenic fungi are concentrated in the following orders: Hypocreales (various genera), Onygenales (Ascosphaera genus), Entomophthorales, and Neozygitales (Entomophthoromycota).(AU)

Os fungos entomopatogênicos são importantes agentes de controle biológico em todo o mundo e têm sido objeto de intensa pesquisa por mais de 100 anos, infectando artrópodes na natureza e podendo ocorrer em níveis enzoóticos ou epizoóticos em suas populações de hospedeiros. O seu mecanismo de infecção envolve a fixação do esporo à cutícula do inseto, seguido da germinação, penetração da cutícula e disseminação interna no inseto. As linhagens dos fungos entomopatogênicos estão concentradas nas ordens: Hypocreales (vários gêneros), Onygenales (gênero Ascosphaera), Entomophthorales e Neozygitales (Entomophthoromycota).(AU)

Aflatoxin M1 contamination in grated parmesan cheese marketed in Rio de Janeiro - Brazil

This study aimed to determine the occurrence of AFM1 contamination in the samples of grated parmesan cheese marketed in the Metropolitan Region of Rio de Janeiro -Brazil. Thirty samples representing 10 major brands marketed in the region were analyzed by High Performance Liquid Chromatography with fluorescence detection (HPLC-FLD) after purification with immunoaffinity column. The method showed recovery values within the range of 70-90%, with RSD lower than 15% and limits of detection and quantification below the maximum level allowed by the European Commission for the presence of AFM1 in cheeses. The mycotoxin was identified in 18 (60%) of the grated cheese samples tested. The highest value corresponded to 0.69 ± 0.02 µg/kg and the mean for all the analyzed samples was 0.16 µg/kg. All the samples were lower than the limit established by the Brazilian legislation (2.5 µg/kg) for AFM1 in cheeses in general. However, eight samples (26.7%) presented AFM1 levels above the tolerance limit of 0.25 µg/kg adopted by the European Commission. These results indicated that AFM1 levels in the grated cheese consumed in Rio de Janeiro -Brazil were relatively high and it could provide a potential hazard for the public health.

Selection of cellulolytic fungi isolated from diverse substrates

The aim of the present work was to select filamentous fungi isolated from diverse substrates to obtain the strains with potential to produce the hydrolytic enzymes. From a total of 215 strains, seven strains from the soils, six from the plants and one from sugarcane bagasse were selected and identified as belonging to the Trichoderma, Penicillium and Aspergillus genera. The best hydrolytic activities obtained by semi-solid fermentation using these strains were approximately: 35; 1; 160; 170 and 120 U/gdm (CMCase, FPase, β-glucosidase, xylanase and polygalacturonase, respectively), demonstrating their potential to synthesize the enzymes compared with the results reported in the literature.

Seleção de isolados de Metarhizium anisopliae e Beauveria bassiana patogênicos a soldados de Atta bisphaerica e Atta sexdens rubropilosa em condições de laboratório / Selection of Metarhizium anisopliae and Beauveria bassiana isolates pathogenic to Atta bisphaerica and Atta sexdens rubropilosa soldiers under laboratory conditions

As formigas do gênero Atta são pragas importantes de diversas culturas agrícolas, pastagens e reflorestamentos. Os fungos entomopatogênicos estão entre os fatores naturais de mortalidade dessas formigas e por isso apresentam potencial para serem usados no controle biológico dessa praga. O presente trabalho teve como objetivo selecionar isolados de Metarhizium anisopliae e Beauveria bassiana patogênicos a soldados de Atta bisphaerica e Atta sexdens rubropilosa em condições de laboratório. Para a avaliação da patogenicidade, foram utilizados oito isolados de M. anisopliae e seis de B. bassiana. O experimento foi conduzido em DIC, sendo a parcela composta por um grupo de 10 soldados por espécie de formiga, sendo utilizadas três repetições por tratamento. Para cada isolado, três grupos de soldados foram pulverizados com suspensão de 1,0 x 10(8) conídios ml-1 e mantidos em câmara úmida (25±1°C, 80±1 por cento de UR e no escuro) sem alimentação, sendo a mortalidade verificada diariamente. Dos 14 isolados testados, quatro de M. anisopliae e quatro de B. bassiana foram patogênicos aos soldados de ambas as espécies de formigas. A virulência foi avaliada para os isolados que causaram mortalidade igual ou maior a 50 por cento. Para cada isolado, suspensões contendo 1,0 x 10(6) a 1,0 x 10(11) conídios ml-1 foram pulverizadas sobre três grupos de 10 soldados e igualmente acondicionados como no teste de patogenicidade. A porcentagem de mortalidade foi calculada a cada 24 horas para determinação do TL50. O isolado ENA04 de M. anisopliae foi mais patogênico, causando mais de 80 por cento de mortalidade nos primeiros três dias após a inoculação, apresentou maior capacidade de esporular nos cadáveres dos soldados e foi o mais virulento para os soldados de A. bisphaerica, com um TL50 de 1,15 dias. Todos os isolados patogênicos aos soldados de A. sexdens rubropilosa foram igualmente virulentos.

The ants of the genus Atta are important pests of several crops, pastures and planted forests. The entomopathogenic fungi are among the natural mortality factors of these ants and because of that they have potential to be use in biological control of this pest. The present research aimed to select isolates of Metarhizium anisopliae and Beauveria bassiana pathogenic to soldiers of Atta sexdens rubropilosa and Atta bisphaerica under laboratory conditions. To evaluate the pathogenicity, eight isolates of M. anisopliae and six of B. bassiana were used. The experiment was carried out in a completely randomized design, being the plots composed by a group of ten soldiers per each ant species, and three replicates per treatment. For each isolate, three groups of soldiers were sprayed with conidial suspensions containing 1.0 x 10(8) conidia ml-1, kept in moist chamber (25±1°C, 80 1 percent of RH in the dark) without food, and the mortality was evaluated every day. From the 14 isolates tested, four of M. anisopliae and four of B. bassiana were pathogenic to soldiers of the both ant species. The virulence was evaluated to the isolates that caused mortality equal or higher than 50 percent. For each isolate, suspensions containing 1.0 x 10(6) to 1.0 x 10(11) conidia ml-1 were sprayed on three groups of ten soldiers, and equally kept as on the pathogenicity test. The mortality percentage was calculated each 24 hours for determining TL50. The isolate ENA04 of M. anisopliae was the most pathogenic, causing more than 80 percent of mortality in the first three days after inoculation, showed higher capacity of spore production on ant cadavers, and was the most virulent to the soldiers of A. bisphaerica, with a TL50 of 1.15 days. All pathogenic isolates of A. sexdens rubropilosa soldiers were equally virulent.