Molecular detection and caracterization of Anaplasma spp. in sheep

• Grant number: 22/06526-3
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: July 01, 2022
• End date: June 30, 2023
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Marcos Rogério André
• Grantee: Laís Engler
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

Anaplasmosis is a tick-borne disease that affects sheep and other animal species and is caused by ±-proteobacteria of the genus Anaplasma spp. Little is known about the occurrence and genetic diversity of Anaplasma species that parasitize sheep in Brazil. Sheep anaplasmosis has high relevance for the production of these small ruminants, generating economic losses, in addition to the fact that some species of Anaplasma may have zoonotic potential. The present study aims to verify the occurrence and molecular identity of Anaplasma spp. in DNA samples extracted from the blood of 200 sheep in the cities of Guarapuava, in the state of Paraná, Jaboticabal, in the state of São Paulo, and in Campo Grande, in the state of Mato Grosso do Sul. Sheep blood samples will be subjected to DNA extraction and conventional PCR assays for the endogenous gene glyceraldehyde-3-phosphate dehydrogenase (gapdh). Positive samples will be submitted to a conventional PCR assay for Anaplasma spp. based on the 16S rRNA gene and quantitative real-time PCR (qPCR) for Anaplasma marginale based on the msp1² gene. Positive samples will be subjected to additional PCR assays for molecular characterization based on distinct gene fragments (msp4, msp5 and groEL). The amplicons will be purified and sequenced using an automated technique based on the dideoxynucleotide chain termination method. The sequences obtained will be submitted to phylogenetic inferences to position them within an evolutionary relationship with species and genotypes of Anaplasma spp. previously detected in different parts of the world. The present work will contribute to the understanding of the molecular identity of Anaplasma spp. in sheep in Brazil.(AU)