Isolation and genotyping of Toxoplasma gondii in sheep meat from Northern Parana state, Brazil

Maria Paula de Carvalho Ewald; Fernando Pietrini Soufen; Luiz Daniel de Barros; João Pedro Sasse; Fernando Rodrigues; João Luis Garcia

doi:10.17696/2318-3691.31.01.2025.265

Authors

Maria Paula de Carvalho Ewald Laboratório de Protozoologia Animal, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina – UEL, Londrina, PR, Brasil; Ministério da Agricultura, Pecuária e Abastecimento (MAPA) https://orcid.org/0009-0003-0185-4278
Fernando Pietrini Soufen Laboratório de Protozoologia Animal, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina – UEL, Londrina, PR, Brasil https://orcid.org/0009-0002-6967-1649
Luiz Daniel de Barros Departamento de Medicina Veterinária, Universidade Federal de Lavras – UFLA, MG, Brasil https://orcid.org/0000-0002-8560-3948
João Pedro Sasse Laboratório de Protozoologia Animal, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina – UEL, Londrina, PR, Brasil https://orcid.org/0000-0002-4334-1759
Fernando Rodrigues Universidade Estadual de Londrina https://orcid.org/0000-0003-0786-0882
João Luis Garcia Laboratório de Protozoologia Animal, Departamento de Medicina Veterinária Preventiva, Universidade Estadual de Londrina – UEL, Londrina, PR, Brasil https://orcid.org/0000-0003-1826-7582

DOI:

https://doi.org/10.17696/2318-3691.31.01.2025.265

Keywords:

Toxoplasmosis, Apicomplexa

Abstract

Introduction: Toxoplasmosis is caused by Toxoplasma gondii, an intracellular protozoan that can infect humans and all warm-blooded animals. Objective: The present study aimed to isolate and genetically characterize T. gondii in sheep meat. Methods: The study was conducted in a sheep slaughterhouse, and samples of masseter muscles, tongue, heart, diaphragm, and blood were collected from 50 sheep. The sheep sera were subjected to an indirect immunofluorescence assay (IFA) to detect anti-Toxoplasma gondii IgG antibodies. The cutoff point used was 1:64. Tissue samples from seropositive animals were subjected to mouse bioassay. Inoculating mice determined the virulence of isolated strains. Genotyping was performed using multilocus PCR-RFLP with 11 genetic markers. Kaplan-Meier was used to build a survival curve. Results: IFAT showed 12% (6/50) of seropositivity, and the antibody titers were 64 (4/50, 8%) and 256 (2/ 50, 4%). The isolates were named TgShBrPr01 and TgShBrPr02. Genetic characterization of the isolate TgShBrPr01 showed the genotype ToxoDB #183. The isolated TgShBrPr02 did not amplify all markers; however, the available typing data indicate a new genotype. The survival curve of TgShBrPr01 showed a high virulence strain and killed all the Swiss Webster mice infected with 10²and 10³ T. gondii tachyzoites. Conclusion: This study isolated two new atypical (nonclonal) T. gondii strains from sheep, underscoring the potential risk of this transmission source to humans, especially when meat is consumed raw or undercooked.

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Isolation and genotyping of Toxoplasma gondii in sheep meat from Northern Parana state, Brazil

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