MOLECULAR DETECTION AND CHARACTERIZATION OF HEMOPLASMAS AND Bartonella spp. IN NON-HUMAN PRIMATES IN THE STATES OF SÃO PAULO AND MATO GROSSO DO SUL

• Grant number: 24/22292-8
• Support Opportunities: Scholarships in Brazil - Master
• Start date: June 01, 2025
• End date: August 31, 2026
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Marcos Rogério André
• Grantee: Leticia Colovatti Mariano
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

Hemoplasmas (hemotropic mycoplasmas) are pleomorphic, obligate parasitic bacteria that inhabit the surface of erythrocytes. Bartonella spp. are facultative intracellular bacteria that infect erythrocytes, endothelial cells, and various other cell types. Both are parasites of multiple mammal species, including humans, and can cause clinical manifestations ranging from mild to severe, depending on the pathogen species and the immune status of the vertebrate host. Wild animals act as reservoirs and amplifiers for various pathogens. In recent decades, with the expansion of rural communities and deforestation, non-human primates (NHPs) have had closer contact with humans, facilitating the exchange of pathogenic agents. This study aims to investigate the occurrence and molecular characterization of hemotropic mycoplasmas and Bartonella species in blood, clot, and spleen samples from NHPs across the states of São Paulo and Mato Grosso do Sul. A total of 211 biological samples from 10 NHP species will be analyzed. DNA will be extracted from the biological samples and subjected to qPCR assays for Bartonella spp. (nuoG) and PCR for hemoplasmas (16S rRNA gene). Positive samples in the qPCR assays for Bartonella spp. will undergo further PCR tests targeting seven molecular markers (gltA, ribC, rpoB, ftsZ, groEL, pap-31, and nuoG). Additionally, blood samples from NHPs collected at the Associação Mata Ciliar in Jundiaí will undergo liquid culture using the enriched Bartonella-Alphaproteobacteria Growth Medium (BAPGM). After incubation, aliquots of the liquid cultures will be extracted for DNA and subjected to qPCR for Bartonella spp., while part will be plated on chocolate agar. Colonies suggestive of Bartonella spp. isolated on chocolate agar will be subjected to DNA extraction, qPCR, and PCR assays for molecular characterization of Bartonella spp. Positive samples in the hemoplasma screening PCR will be further tested for the complete 16S rRNA gene and 23S rRNA gene. The amplicons will be purified and sequenced using the Sanger method, with molecular identity investigated through BLASTn and phylogenetic analysis. This study will contribute to a better understanding of the diversity of hemoplasmas and Bartonella spp. in NHPs in Brazil. (AU)