Serological, epidemiological and genetic profile of Toxoplasma gondii from reproductive tissues of dogs and cats

• Grant number: 20/03972-7
• Support Opportunities: Regular Research Grants
• Duration: March 01, 2022 - February 29, 2024
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Cinara de Cássia Brandão
• Grantee: Cinara de Cássia Brandão
• Host Institution: Faculdade de Medicina de São José do Rio Preto (FAMERP). Secretaria de Desenvolvimento Econômico (São Paulo - Estado). São José do Rio Preto , SP, Brazil
• Associated researchers: Fernando Henrique Antunes Murata ; Luiz Carlos de Mattos
• Associated scholarship(s): 22/08270-6 - Genotyping of Toxoplasma gondii isolated in reproductive tissues of dogs and cats, BP.TT

Abstract

Toxoplasmosis is a disease caused by the protozoan Toxoplasma gondii. The presence of cats is a determining factor for the occurrence of toxoplasmosis, which together with dogs, serve as indicators of environmental contamination by T. gondii, especially due to their proximity to humans. Demonstrations of this parasite in animal reproductive tissues (testicles, ovaries and uterus) are inconclusive and still need further clarification. Such tissues are excellent biological materials for analysis in order to determine the diagnosis of the infection, as well as to know the prevalence of toxoplasmosis in areas with a higher rate of occurrence of T. gondii. The objectives of the study are: 1. to determine the risk of taking anti-T. gondii (IgG and IgM) in cats and dogs during sterilization at the Zoonoses Center of São José do Rio Preto; 2. isolate T. gondii from reproductive tissues through mouse bioassay and cell culture; 3. genotype, by means of microsatellites and PCR-RFLP and next-generation sequencing (NGS), the T. gondii grantees chose; 4. Geocode the animal addresses in order to determine risk areas. Blood samples will be collected by puncture of the jugular, cephalic or saphenous vein with the animals under anesthesia, to obtain serum and serological evaluation by modified agglutination test (MAT). Testis, uterus and ovary macerates will be used in bioassays with Balb C mice of both sexes, to obtain the concession of T. gondii. Those obtained will be obtained for genomic characterization by PCR-RFLP, using 11 distinct markers: SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. Geocoding uses epidemiological data from animals (sex, age and neighborhood in which they live) to determine potential areas of infection and disease prevalence, as well as environmental contamination. The results contributed to the characterization of T. gondii strains in domesticated animals as well as in homeless ones, favoring the identification and characterization of environmental infection and risks. The genomic characterization of those required to help identify indicators of T. gondii virulence, as well as establish possible relationships with the infection in humans, including caregivers and guardians of homeless animals. (AU)