Evaluation of experimental Toxoplasma gondii (Nicolle and Manceaux, 1909) infection in pigs by bioassay in mice and polymerase chain reaction

  • Lucia Eiko Oishi Yai Prefeitura do Munícipio de São Paulo, Centro de Controle de Zoonoses, São Paulo, SP
  • Manoella Compostrini Barreto Vianna Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Medicina Veterinária Preventiva e Saúde Animal, São Paulo, SP
  • Rodrigo Martins Soares Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Medicina Veterinária Preventiva e Saúde Animal, São Paulo, SP
  • Adriana Cortez Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Medicina Veterinária Preventiva e Saúde Animal, São Paulo, SP
  • Roberta Lemos Freire Universidade Estadual de Londrina, Centro de Ciências Agrárias, Londrina, PR
  • Leonardo José Richtznhain Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Medicina Veterinária Preventiva e Saúde Animal, São Paulo, SP
  • Solange Maria Gennari Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, Departamento de Medicina Veterinária Preventiva e Saúde Animal, São Paulo, SP
Keywords: Toxoplasma gondii, Pig, NestedPCR, Bioassay, Diagnosis

Abstract

The aim of the present experiment was to standardize a nested polymerase chain reaction (nPCR) for the detection of Toxoplasma gondii in tissues of experimentally infected pigs and to compare the performance of nPCR with the standard isolation technique, the bioassay in mice. Comparison between the two methods was done testing eight 4 month-old pigs orally inoculated with 5 x 10(4) oocysts of Toxoplasma gondii (AS-28 strain) and three non-infected pigs at the same age, kept as control. All animals were euthanatized 47 days after infection and samples of brain, heart, tongue and retina were collected from each animal for analysis by nPCR and bioassay in mice. By using the bioassay, Toxoplasma gondii was detected in 4 infected pigs, being two in the retina, one in the heart and one in the tongue. Toxoplasma gondii DNA was detected in five of the inoculated pigs, being: three in the tongue, two in the brain and heart and one in retina. The detection threshold of the nPCR on mouse brain suspension artificially infected with the RH strain of Toxoplasma gondii was 10 tachyzoites/ml. Although both techniques were unable to detect the parasite in all infected pigs, nPCR showed better performance as it was accomplished in a shorter period of time. When used concurrently, both techniques detected the agent in seven infected animals. The only way to increase sensitivity of either method is to increase the amount of tissue to be examined.

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Published
2003-01-01
How to Cite
Yai, L., Vianna, M., Soares, R., Cortez, A., Freire, R., Richtznhain, L., & Gennari, S. (2003). Evaluation of experimental Toxoplasma gondii (Nicolle and Manceaux, 1909) infection in pigs by bioassay in mice and polymerase chain reaction. Brazilian Journal of Veterinary Research and Animal Science, 40(3), 227-234. https://doi.org/10.1590/S1413-95962003000300010
Section
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