ISOLATION AND GENOTYPING OF Bartonella spp. IN DOMESTIC AND WILD RESERVOIR MAMMALS IN BRAZIL

Abstract

Bartonellosis, an emerging zoonosis, is caused by Gram-negative bacteria belonging to the genus Bartonella. This group arthropod-borne pathogens comprises agents highly adapted to a wide variety of hosts, causing different clinical manifestations depending on the species and host involved in the infection. Although rodents, bats and domestic felines are considered important reservoirs for Bartonella species worldwide, few or no studies have been conducted in Brazil regarding the isolation and genotypic diversity of Bartonella spp. in these mammal species and their respective ectoparasites. The present study aims to isolate and characterize, using microbiological and molecular methods, members of the Bartonella genus in blood/tissue samples of synanthropic rodents, bats, domestic cats and their ectoparasites in selected localities in Brazil. For this purpose, mammals blood and/or tissues and ectoparasites DNA samples will be screened for Bartonella by qPCR based on the nuoG gene. In addition, whole blood samples from mammals will be submitted to the liquid enrichment culture followed by solid culture. Positive cultures as well as DNA samples from ectoparasites and blood/tissues will be submitted to additional cPCR assays for Bartonella spp. based on the 16S rRNA, gltA, rpoB, ftsZ, pap-31, groEL and ribC genes and the 16S- 23S rRNA (ITS) genic fragments. Bartonella amplicons based on different genic regions (Multi- Locus Sequencing Typing - MLST) obtained from ectoparasites and blood/tissue positive will be cloned. Samples will be sequenced by the Sanger method and the amplified sequences will be compared with those previously deposited in Genbank and later submitted to phylogenetic inference using the Maximum Likelihood and Bayesian methods. In addition, Median Network analysis will be conducted. The results of the present study will contribute to the elucidation of the genetic diversity of Bartonella species circulating between mammalian reservoirs and their respective ectoparasites in our country. (AU)