One of the interesting features of Anticarsia gemmatalis multiple nucleopolyhedrovirus isolate 2D (AgMNPV-2D) genome is the absence of chitinase (chiA) and cathepsin (v-cath) genes. This characteristic may be responsible for the lack of liquefaction and melanization in A. gemmatalis larvae killed by AgMNPV-2D infection. This study aimed to test the hypothesis that CHIA and V-CATH proteins from Choristonera fumiferana DEF multiple nucleopolyhedrovirus (CfDEFNPV) are able to liquefy and melanize the cuticle of A. gemmatalis larvae infected by a recombinant AgMNPV containing chiA and v-cath genes inserted in its genome. A fragment from the CfDefNPV genome containing chiA and v-cath genes was inserted into the genome of AgMNPV-2D. The recombinant virus (vAgp2100Cf.chiA/v-cath) was purified and used to infect insect cells and larvae. Transcripts of v-cath and chiA genes were detected along the infection of insect cells by qRT-PCR, from early to late phases of infection. The analysis of A. gemmatalis larvae killed by vAgp2100Cf.chiA/v-cath infection confirmed the hypothesis proposed. The vAgp2100Cf.chiA/v-cath showed higher insecticidal activity against third instar A. gemmatalis larvae when compared to AgMNPV-2D. The mean time to death was also lower for the vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D at 10 days post infection. Occlusion body production was higher in A. gemmatalis larvae infected with vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D. Enzyme assays showed higher chitinase and cysteine protease activities in insect cells and insects infected with vAgp2100Cf.chiA/v-cath when compared to AgMNPV-2D. The introduction of chiA and v-cath genes into the genome of AgMNPV improves its insecticidal activity against A. gemmatalis larvae and this recombinant virus could be used as an alternative to the wild type virus to control this important insect pest.

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Qualidade microbiológica de queijo Minas frescal, artesanal, comercializados em feiras livres do Distrito FederalMicrobiological quality of Minas Frescal Cheese, handcrafted, marketed at free fairs of the Federal District

Análise microbiológica de água de coco comercializadas em garrafas plásticas dentro do Distrito Federal Microbiological analysis of coconut water sold in plastic bottles within the Federal District

ResumoAspergillus e Penicillium são microrganismos capazes de provocar complicações clínicas consideráveis, principalmente em pacientes imunossuprimidos. A análise molecular é uma estratégia importante para a identificação rápida e precisa desses agentes patogênicos. Por isso, o objetivo deste trabalho é descrever padrões de marcadores RAPD específicos para a identificação de isolados de Aspergillus e Penicillium causadores de doenças de interesse médico. A análise morfológica revelou que 14 isolados eram do gênero Aspergillus, 7 eram Penicillium, 1 correspondente a Fusarium e 1 isolado não foi identificado. A análise por RAPD com 10(dez) iniciadores produziu fragmentos de DNA característicos para cada gênero.O dendrograma revelou grupos de isolados dentro de cada gênero de fungo de acordo com perfis de marcadores específicos, identificando isolados muito semelhantes que apresentaram alto coeficiente de similaridade. Esses dados fornecem opções para o desenvolvimento de oligonucleotídios de PCR para a identificação de isolados específicos que ocorrem no ambiente médico-hospitalar.Palavras-chave: Análise de DNA. Aspergillus. Penicillium. Polimorfismo.