DOI QR코드

DOI QR Code

Isolation and Genotyping of Toxoplasma gondii in Brazilian Dogs

  • Received : 2016.10.11
  • Accepted : 2017.04.19
  • Published : 2017.06.30

Abstract

Strains of Toxoplasma gondii in Brazil are highly genetically diverse compared to strains from North America and Europe. Dogs are epidemiologically important because they act as sentinels for T. gondii infections in humans and are good indicators of environmental contamination. The aim of this study was to isolate and genetically characterize T. gondii strains from tissues of naturally infected Brazilian dogs. For this study, 21 blood samples were collected from dogs at the Zoonosis Control Centers of $Ilh{\acute{e}}us$ and Itabuna cities, Bahia, Brazil. The sera were examined for T. gondii antibodies using the indirect hemagglutination test. Brains and hearts of seropositive dogs were bioassayed in mice to isolate and characterize T. gondii parasites by PCR-RFLP using 10 genetic markers (SAG1, newSAG2, SAG3, BTUB, c22-8, c29-2, GRA6, PK1, APICO, and L358). However, T. gondii was isolated from only 4 (57.1%) dogs, designated TgDgBr6, 13, 17, and 21. All strains were virulent, causing clinical changes (rough hair coat, lethargy, and abdominal distention) and the death of all mice within 8-20 days after inoculation. Genetic analysis of these 4 T. gondii isolates revealed 4 distinct genotypes with different clonal lineage combinations (types I, II, and III) and 2 atypical alleles. Using PCR-RFLP with several markers, this study contributes to evaluations of the genetic diversity of strains circulating in Brazil.

Keywords

References

  1. Dubey JP, Beattie CP. Toxoplasmosis of Animals and Man. Florida, USA. CRC. 1988, pp 1-220.
  2. Dubey JP, Zarnke R, Thomas NJ, Wong SK, Van Bonn W, Briggs M, Davis JW, Ewing R, Mense M, Kwok OC, Romand S, Thulliez P. Toxoplasma gondii, Neospora caninum, Sarcocystis neurona, and Sarcocystis canis-like infections in marine mammals. Vet Parasitol 2003; 116: 275- 296. https://doi.org/10.1016/S0304-4017(03)00263-2
  3. Ribeiro LA, Santos LK, Brito PA Jr, Maciel BM, Da Silva AV, Albuquerque GR. Detection of Toxoplasma gondii DNA in Brazilian oysters (Crassostrea rhizophorae). Genet Mol Res 2015; 14: 4658-4665. https://doi.org/10.4238/2015.May.4.25
  4. Dubey JP, Lago EG, Gennari SM, Su C, Jones JL. Toxoplasmosis in humans and animals in Brazil: high prevalence, high burden of disease and epidemiology. Parasitology 2012; 139: 1375-1424. https://doi.org/10.1017/S0031182012000765
  5. Dubey JP. Toxoplasmosis-a waterborne zoonosis. Vet Parasitol 2004; 126: 57-72. https://doi.org/10.1016/j.vetpar.2004.09.005
  6. da Silva RC, de Lima VY, Tanaka EM, da Silva AV, de Souza LC, Langoni H. Risk factors and presence of antibodies to Toxoplasma gondii in dogs from the coast of Sao Paulo State, Brazil. Pesq Vet Bras 2010; 30: 161-166. https://doi.org/10.1590/S0100-736X2010000200011
  7. Yan C, Fu LL, Yue CL, Tang RX, Liu YS, Lv L, Shi N, Zeng P, Zhang P, Wang DH, Zhou DH, Zhu XQ, Zheng KY. Stray dogs as indicators of Toxoplasma gondii distributed in the environment: the first report across an urban-rural gradient in China. Parasit Vectors 2012; 5: 5. https://doi.org/10.1186/1756-3305-5-5
  8. Bahia-Oliveira LM1, Jones JL, Azevedo-Silva J, Alves CC, Orefice F, Addiss DG. Highly endemic, waterborne toxoplasmosis in north Rio de Janeiro state, Brazil. Emerg Infect Dis 2003; 9: 55-62. https://doi.org/10.3201/eid0901.020160
  9. Pena HF, Gennari SM, Dubey JP, Su C. Population structure and mouse-virulence of Toxoplasma gondii in Brazil. Int J Parasitol 2008; 38: 561-569. https://doi.org/10.1016/j.ijpara.2007.09.004
  10. Su C, Khan A, Zhou P, Majumdar D, Ajzenberg D, Darde ML, Zhu XQ, Ajioka JW, Rosenthal BM, Dubey JP, Sibley LD. Globally diverse Toxoplasma gondii isolates comprise six major clades originating from a small number of distinct ancestral lineages. Proc Natl Acad Sci USA 2012; 109: 5844-5849. https://doi.org/10.1073/pnas.1203190109
  11. Bezerra RA, Carvalho FS, Guimaraes LA, Rocha DS, Maciel BM, Wenceslau AA, Lopes CW, Albuquerque GR. Genetic characterization of Toxoplasma gondii isolates from pigs intendend for human consumption in Brazil. Vet Parasitol 2012; 189: 153-161. https://doi.org/10.1016/j.vetpar.2012.04.036
  12. Clementino-Andrade MM, Pinheiro BV, Cunha MM, Carneiro AC, Andrade Neto VF, Vitor RW. New genotypes of Toxoplasma gondii obtained from farm animals in Northeast Brazil. Res Vet Sci 2013; 94: 587-589. https://doi.org/10.1016/j.rvsc.2013.01.006
  13. Maciel BM, Moura RL, Carvalho FS, Costa EA, Albuquerque GR. Identification and genetic characterization of a new Brazilian genotype of Toxoplasma gondii from sheep intended for human consumption. Parasitol Int 2014; 6: 567-570.
  14. Silva LA, Andrade RO, Carneiro AC, Vitor RW. Overlapping Toxoplasma gondii genotypes circulating in domestic animals and humans in shoutheastern Brazil. PLoS One 2014; 9: 1-7.
  15. Ajzenberg D, Banuls AL, Su C, Dumetre A, Demar M, Carme B, Darde ML. Genetic diversity, clonality and sexuality in Toxoplasma gondii. Int J Parasitol 2004; 34: 1185-1196. https://doi.org/10.1016/j.ijpara.2004.06.007
  16. Carlos RSA, Albuquerque GR, Bezerra RA, Sicupira PML, Munhoz AD, Lopes CWG. Ocorrencia de anticorpos anti-Toxoplasma gondii e principais fatores de risco associados a infeccao canina na regiao de Ilheus-Itabuna, estado da Bahia. Rev Bras Med Vet 2010; 32: 115-121 (in Portuguese).
  17. Dubey JP. Refinement of pepsin digestion method for isolation of Toxoplasma gondii from infected tissues. Vet Parasitol 1998; 74: 75-77. https://doi.org/10.1016/S0304-4017(97)00135-0
  18. Homan WL, Vercammen M, De Braekeleer J, Verschueren H. Identification of a 200 to 300 fold repetitive 529 pb DNA fragment in Toxoplasma gondii, and its use for diagnostic and quantitative PCR. Int J Parasitol 2000; 30: 69-75. https://doi.org/10.1016/S0020-7519(99)00170-8
  19. Su C, Shwab EK, Zhou P, Zhu XQ, Dubey JP. Moving towards an integrated approach to molecular detection and identification of Toxoplasma gondii. Parasitology 2010; 137: 1-11.
  20. Bezerra RA, Gine GA, Maciel BM, Gaiotto FA. Albuquerque GR. Identification and genetic characterization of Toxoplasma gondii in free-ranging bristle-spined porcupine (Chaetomys subspinosus), a threatened arboreal mammal from the Brazilian Atlantic Forest. Parasit Vectors 2015; 8: 277. https://doi.org/10.1186/s13071-015-0882-6
  21. Bezerra RA, Carvalho FS, Guimaraes LA, Rocha DS, Silva FL, Wenceslau AA, Albuquerque GR. Comparison of methods for detection of Toxoplasma gondii in tissues of naturally exposed pigs. Parasitol Res 2012; 110: 509-514. https://doi.org/10.1007/s00436-011-2514-1
  22. Dubey JP, Hill DE, Rozeboom DW, Rajendaran C, Choudhary S, Ferreira LR, Kwok OC, Su C. High prevalence and genotypes of Toxoplasma gondii isolated from organic pigs in northern USA. Vet Parasitol 2012; 188: 14-18. https://doi.org/10.1016/j.vetpar.2012.03.008
  23. Dubey JP, Rajendran C, Ferreira LR, Martins J, Kwok OC, Hill DE, Villena I, Zhou H, Su C, Jones JL. High prevalence and genotypes of Toxoplasma gondii isolated from goats, from a retail meat store, destined for human consumption in the USA. Int J Parasitol 2011; 41: 827-833. https://doi.org/10.1016/j.ijpara.2011.03.006
  24. El Behairy AM, Choudhary S, Ferreira LR, Kwok OC, Hilali M, Su C, Dubey JP. Genetic characterization of viable Toxoplasma gondii isolates from stray dogs from Giza, Egypt. Vet Parasitol 2013; 193: 25-29. https://doi.org/10.1016/j.vetpar.2012.12.007
  25. Dubremetz JF, Lebrun M. Virulence factors of Toxoplasma gondii. Microbes Infect 2012; 14: 1403-1410. https://doi.org/10.1016/j.micinf.2012.09.005
  26. Dubey JP, Gennari SM, Sundar N, Vianna MC, Bandini LM, Yai LE, Kwok OC, Su C. Diverse and atypical genotypes identified in Toxoplasma gondii from dogs in Sao Paulo, Brazil. J Parasitol 2007; 93: 60-64. https://doi.org/10.1645/GE-972R.1
  27. Langoni H, Matteucci G, Medici B, Camossi LG, Richini-Pereira VB, Silva RC. Detection and molecular analysis of Toxoplasma gondii and Neospora caninum from dogs with neurological disorders. Rev Soc Bras Med Trop 2012; 45: 365-368. https://doi.org/10.1590/S0037-86822012000300016
  28. Soares RM. Caracterizacao molecular de Toxoplasma gondii. Rev Bras Parasitol Vet 2004; 13: 17-19 (in Portuguese).
  29. da Silva AV, da Silva RC, Zamprogna TO, Lucas TM. Toxoplasma gondii in swine with emphasis in Brazilian contribution. Sci Med 2010; 20: 120-130.
  30. Darde ML. Toxoplasma gondii, “new” genotypes and virulence. Parasite 2008; 15: 366-371. https://doi.org/10.1051/parasite/2008153366

Cited by

  1. Dogs ( Canis familiaris ) as Sentinels for Human Infectious Disease and Application to Canadian Populations: A Systematic Review vol.5, pp.4, 2017, https://doi.org/10.3390/vetsci5040083
  2. Virulence of atypical Toxoplasma gondii strains isolated in French Guiana in a murine model vol.26, pp.None, 2017, https://doi.org/10.1051/parasite/2019048
  3. Clinical Toxoplasmosis in Dogs and Cats: An Update vol.6, pp.None, 2017, https://doi.org/10.3389/fvets.2019.00054
  4. Cytologic detection of Toxoplasma gondii in the cerebrospinal fluid of a dog and in vitro isolation of a unique mouse-virulent recombinant strain vol.33, pp.3, 2017, https://doi.org/10.1177/1040638721996685