ISOLATION, GENOTYPING AND GENOMIC SEQUENCING OF Bartonella spp. IN SMALL WILD MAMMALS AND RUMINANTS IN BRAZIL

Abstract

Around 75% of the emerging diseases evolve zoonoses, which have in its majority wild animals as sources of infection and are primarily transmitted by arthropod vectors. Among them, bartonelloses diseases are caused by pathogens belonging to the Bartonella genus, which comprise fastidious Gram-negative microorganisms highly specialized in parasitizing mammals. Most of mammalian species belongs to the Rodentia Order, which act as important reservoirs for various pathogens. Even though few studies aiming the characterization of genotypic Bartonella in rodents and marsupials have been performed in Brazil, those related to small ruminants are absent so fat. The present study aims to isolate and investigate the genetic diversity of Bartonella spp. in rodents, marsupials sampled in the Pantanal region of the state of Mato Grosso do Sul, in the Nhumirim farm of Embrapa Pantanal / CPAP and Nhecolândia region. In addition, blood samples will be collected from sheep in some localities in the states of Mato Grosso do Sul, São Paulo and Paraná. Blood samples and ectoparasites collected from the sampled animals will be submitted to solid chocolate agar culture for Bartonella isolation. Positive cultures, as well as DNA samples extracted from ectoparasites and blood or tissues, will be submitted to real time and conventional PCR assays based on different target genes (nuoG, pap-31, gltA, rpoB, ftsZ, groEL and ribC). Genic cloning of the amplified fragments will be performed in order to analyze the diversity of genotypes circulating in a single host (vertebrate or ectoparasite). The Whole Genome Sequencing technique will also be performed aiming at determining the complete genome of the genotype isolated from rodents and/or marsupials. Moreover, the obtained sequences will be submitted to phylogenetic inferences using the Maximum Likelihood and Bayesian methods. The genotypes network will be performed using Median Network. Therefore, the present study will contribute to the study of the genetic diversity of Bartonella spp. in Brazilian wildlife.