Molecular detection and characterization of Bartonella spp. in sandflies and biting midges in the Brazilian Amazon.

• Grant number: 22/16085-4
• Support Opportunities: Scholarships in Brazil - Master
• Start date: April 01, 2023
• End date: March 31, 2025
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Pathology
• Principal Investigator: Marcos Rogério André
• Grantee: Paulo Vitor Cadina Arantes
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

The genus Bartonella spp. encompasses facultative intracellular Gram-negative aerobic bacteria parasitic on erythrocytes and endothelial cells of various mammals, including humans. They are etiological agents of several diseases, such as trench fever, cat scratch disease, bacillary angiomatosis, peliosis hepatis, endocarditis, among others. Despite the vast occurrence in mammals around the world, there are few studies about the genetic diversity of Bartonella spp. in arthropods and little is known about potential vectors. The present study aims to investigate the occurrence and genetic diversity of Bartonella spp. in sandflies and biting midges in the Brazilian Amazon. For this, 600 specimens of dipterans (300 sandflies and 300 biting midges) will be collected in the Amazon National Park, state of Pará. The specimens will be submitted to DNA extraction and initially to conventional PCR based on the cox-1 gene as an endogenous control. Positive samples in that PCR will be submitted to quantitative real-time PCR for Bartonella spp. based on the nuoG gene. Positive samples in qPCR will be submitted to conventional PCR assays based on gltA, ftsZ, pap-31, groEL, rpoB, ribC and intergenic region (ITS) 16-23S rRNA genes. Positive samples will be purified and sequenced by the Sanger method. The phylogenetic positioning of the obtained sequences will be carried out through the construction of dendograms. The present study will contribute to the study of the diversity of Bartonellaceae agents circulating in Nematocera diptera present in the Brazilian territory.