Caracterização das respostas transcricionais e microbiomas de populações naturais do mosquito Aedes aegypti com diferentes níveis de suscetibilidade ao vírus dengue

Dengue is the arbovirosis of greater growth in recent years, reflecting on social and economic impacts due to the high morbidity and mortality rates triggered by infection. Dengue has Aedes aegypti as its primary vector, which is distributed throughout tropical and subtropical regions of the planet. Due to its anthropophilic and haematophagic habit, rapid development and specific behavioral characteristics, it is an excellent dengue virus transmitter. Control measurements are restricted to eliminating the mosquito vector since neither specific treatments nor tetravalent vaccines are yet commercially available. One feature that determines the spread of the disease is the high vector competence of its vector mosquitoes, which has been associated to its genetic factors and to its gut microbiota. The present study evaluated genetic and microbial factors related to vector competence of natural populations of Ae. Aegypti with different levels of susceptibilities to dengue virus serotype 4 (DENV-4). For that, we evaluated the susceptibility to DENV-4 infection of two field collected mosquito populations (Botucatu-SP and Neópolis-SE) through infection assays and relative quantification by qPCR. Differential gene expression and gut microbial diversity analyses were performed using next-generation sequencing (RNA-seq and 16S rRNA gene sequencing) through Illumina HiScanSQ and MiSeq platforms, respectively. The populations presented different susceptibility to DENV-4, of which Botucatu showed higher resistance to infection when compared to Neópolis. Differential gene expression analysis revealed that there was little gene modulation in response to DENV infection in Botucatu, while Neópolis showed consistent modulation on several genes related to immune pathways or digestive processes. When comparing to other studies we found that the Toll immune pathway is being activated in this population after infection. However, the comparison of ...Dengue é a arbovirose de maior crescimento nos últimos anos, repercutindo em impactos sociais e econômicos devido às altas taxas de morbidade e mortalidade desencadeadas pela infecção. O dengue tem como principal vetor o mosquito Aedes aegypti, distribuído em toda a faixa tropical e subtropical do planeta. Por apresentar hábito hematofágico antropofílico, rápido desenvolvimento e características comportamentais específicas, é um excelente transmissor do vírus dengue. Medidas de controle estão restritas à eliminação do mosquito vetor, uma vez que um tratamento específico ou uma vacina que previna simultaneamente a infecção pelos quatro sorotipos deste arbovírus ainda não estão disponíveis à população. Uma característica que determina a disseminação de doenças é a alta competência vetorial de seus mosquitos transmissores, que tem sido associada à fatores genéticos do mosquito bem como à microbiota intestinal do inseto. O presente estudo avaliou fatores genéticos e microbianos relacionados à competência vetorial de populações naturais de Ae. aegypti com diferenças na susceptibilidades ao vírus dengue sorotipo 4 (DENV-4). Para isso, foi avaliada a susceptibilidade à infecção pelo DENV de duas populações naturais deste inseto (Botucatu-SP e Neópolis-SE) através de ensaios de infecção e quantificação relativa por PCR em tempo real. A expressão gênica diferencial, bem como a diversidade microbiana intestinal, foi realizada através do sequenciamento de nova geração (RNA-seq e sequenciamento do gene 16S rRNA) através das plataformas Illumina HiScan™SQ e MiSeq, respectivamente. Verificamos que as populações estudadas apresentam diferenças na susceptibilidades ao DENV-4 onde Botucatu apresentou maior resistência à infecção quando comparada à Neópolis. Pela análise de expressão gênica diferencial verificamos que houve pouca modulação genica na população de Botucatu em resposta a...Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Characterization of Hepatozoon spp. in Leptodactylus chaquensis and Leptodactylus podicipinus from two regions of the Pantanal, state of Mato Grosso do Sul, Brazil

Hepatozoon sp. are parasites that commonly infect frogs and arthropod vectors. This species has variability in the morphological and morphometric characteristics. Due to these variations, the naming of the species is thus impaired and only by visualizing the sporogonic cycle in vector and by molecular studies this problem can be solved. Recently, the use of molecular genetics has helped the species denomination. In this work, we collected 145 frogs (68 Leptodactylus chaquensis and 77 Leptodactylus podicipinus) in different sampling sites, where were found 18 (26.47 %) L. chaquensis and 24 (31.17 %) L. podicipinus parasitized; besides of gamonts, schizogonic forms were also seen in animals organs. The positivity difference between the collection sites for both frog species was not significant (p=0.958). Comparing gamonts found in each species of anuran, we observed differences in morphology. The comparison in the molecular level for L. podicipinus was not possible due to small amount of blood obtained, just L. chaquensis had their parasites DNA sequenced. The amplified and sequenced samples, named HEP1 to HEP10, are presented in the phylogenetic tree as a different branch from other haemogregarines described on other hosts. Therefore, we have seen that, although the morphology and morphometry of the collected parasites at each site showed differences, the sequencing of these samples revealed identical species of Hepatozoon, and different compared to those from GenBank, thereby demonstrating that the species of Hepatozoon in L. chaquensis observed in this study probably represent a new species.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES