Molecular charecterization of coccidia (Toxoplasma gondii, Neospora caninum e Sarcocystis spp.) in the central nervous, ocular and muscular systems of sheep

• Grant number: 25/04006-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: May 01, 2025
• End date: April 30, 2026
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: Gustavo Felippelli
• Grantee: Maria Luísa Pitol Ramos
• Host Institution: Faculdade de Medicina Veterinária (FMVA). Universidade Estadual Paulista (UNESP). Campus de Araçatuba. Araçatuba , SP, Brazil

Abstract

The co-infection of several coccidia is constantly investigated because they have similar organs of predilection, such as the central nervous system, and share the same host class (carnivores) characterising a prey-predator cycle. The aim of this study is to analyse the molecular diagnosis of three parasites from the Sarcocystidae family that share the same intermediate host: sheep (Ovis aries), these protozoa are the following: Toxoplasma gondii, Sarcocystis spp. and Neospora caninum. the first two are zoonotic and the latter is the main cause of reproductive losses in ruminants in general, but with only one serological report in humans. 100 head of sheep donated by a municipal slaughterhouse, will be collected. located in the city of São Manuel, state of São Paulo. The heads will be sectioned to remove the central nervous system, ocular and muscular systems (masseter and tongue) stored at -20°C. DNA will then be extracted from the samples central nervous system, individually: brain, cerebellum and brain stem, eyes (aqueous humor and retina), from the aforementioned muscles; specifically from the tongue and masseter, parasitological diagnosis of Sarcocystis spp. will be carried out. Therefore, a conventional PCR protocol will be carried out on the 18S region for integrated search for DNA from the Sarcocystidae family (Toxoplasma gondii, Sarcocystis spp. and Neosporum caninum). Positive samples will be extracted from the gel using the QIAquick Kit (Qiagen) to purify the PCR amplicons. Genetic sequencing of Sanger gene sequencing will be carried out. The nucleotide sequences sequences will be analysed using the CodonCodeTM Aligner software version 9.0.1, the Bio EditTM Sequence Alignment Editor and Basic Local Aligment Search Tool - BLAST. The data resulting from the PCR protocol will be analysed using the chi-square test with the aid of the Jamovi 2.3.28 and Epi Info 7.2.6.0 to determine the prevalence of the variables, will be considered statistically significant when p ¿ 0.05.