Advertisement

Acta Parasitologica

, Volume 63, Issue 2, pp 397–411 | Cite as

Genotyping and virulence analysis of Toxoplasma gondii isolates from a dead human fetus and dead pigs in Jiangsu province, Eastern China

  • Zhaofeng Hou
  • Yonghua Zhou
  • Dandan Liu
  • Shijie Su
  • Zhenxing Zhao
  • Jinjun Xu
  • Jianping TaoEmail author
Article

Abstract

Toxoplasma gondii is an obligate intracellular parasite with worldwide distribution. Virulence of T. gondii is a multigenic trait. Genetic and virulence data for T. gondii isolates from humans and animals in China have been reported. However, almost all biological materials used for genotyping of T. gondii from humans and pigs were DNA samples prepared from tissues, and T. gondii strains used for virulence analysis were isolated mainly from cats. In this study, one isolate from a dead human fetus was identified as type I (ToxoDB #10) while the two isolates from dead pigs were type Chinese I (ToxoDB #9) with PCR-restriction fragment length polymorphism using 10 markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). Three isolates were comfirmed as virulent strains in mice. By cloning and sequences analysis, all isolates contained a Pvu II restriction site (572–577 bp) in the KHB fragment and five tandem repeats in the 5′ UTR region of SAG1, which were associated with T. gondii virulence. The type Chinese I isolates contained two deletions of 15 and 3 bp at positions 635 to 649 and 658 to 660 in the GRA6, which were correlated with genotype, but not with virulence. To our knowledge, this is the first report on the systematic analysis of murine virulence of type Chinese I strain from pigs, and the associations of sequences of the KHB fragment and SAG1 with virulence of type Chinese I strain. The Chinese I genotype was more closely related to type II strains.

Keywords

Toxoplasma gondii isolates Biological characteristics GRA6 SAG1 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Ajzenberg, D., Yera, H., Marty, P., Paris, L., Dalle, F., Menotti, J., et al. 2009. Genotype of 88 Toxoplasma gondii isolates associated with toxoplasmosis in immunocompromised patients and correlation with clinical findings. Journal of Infectious Diseases, 199, 1155–67. DOI: 10.1086/597477CrossRefGoogle Scholar
  2. Bezerra, R.A., Carvalho, F.S., Guimaraes, L.A., Rocha, D.S., Maciel B.M., Wenceslau, A.A., et al. 2012. Genetic characterization of Toxoplasma gondii isolates from pigs intended for human consumption in Brazil. Veterinary Parasitology, 189, 153–61. DOI: 10.1016/j.vetpar.2012.04.036CrossRefGoogle Scholar
  3. Chen, X., Tan F. 2009. Toxoplasma gondii: past, present and future. Chinese Journal of Parasitology Parasitic Disease, 27, 426–31.Google Scholar
  4. Cong, W., Liu, G.H., Meng, Q.F., Dong, W., Qin, S.Y., Zhang, F.K., et al. 2015. Toxoplasma gondii infection in cancer patients: prevalence, risk factors, genotypes and association with clinical diagnosis. Cancer Letters, 359, 307–13. DOI: 10.1016/j.canlet.2015.01.036CrossRefGoogle Scholar
  5. Dubey, J.P., Hill, D.E., Rozeboom, D.W., Rajendran, C., Choudhary, S., Ferreira, L.R., et al. 2012. High prevalence and genotypes of Toxoplasma gondii isolated from organic pigs in northern USA. Veterinary Parasitology, 188, 14–8. DOI: 10.1016/j.vetpar.2012.03.008CrossRefGoogle Scholar
  6. Dubey, J.P., Van Why, K., Verma, S.K., Choudhary, S., Kwok, O.C., Khan, A., et al. 2014. Genotyping Toxoplasma gondii from wildlife in Pennsylvania and identification of natural recombinants virulent to mice. Veterinary Parasitology, 200, 74–84. DOI: 10.1016/j.vetpar.2013.11.001CrossRefGoogle Scholar
  7. Fazaeli, A., Carter, P.E., Darde, M.L., Pennington T.H. 2000. Molecular typing of Toxoplasma gondii strains by GRA6 gene sequence analysis. International Journal for Parasitology, 30, 637–42CrossRefGoogle Scholar
  8. Fekkar, A., Ajzenberg, D., Bodaghi, B., Touafek, F., Le Hoang, P., Delmas J., et al. 2011. Direct genotyping of Toxoplasma gondii in ocular fluid samples from 20 patients with ocular toxoplasmosis: predominance of type II in France. Journal of Clinical Microbiology, 49, 1513–7. DOI: 10.1128/JCM.02196-10CrossRefGoogle Scholar
  9. Ferreira, I.M., Vidal, J.E., de Mattos Cde, C., de Mattos, L.C., Qu, D., Su, C., et al. 2011. Toxoplasma gondii isolates: multilocus RFLP-PCR genotyping from human patients in Sao Paulo State, Brazil identified distinct genotypes. Experimental Parasitology, 129, 190–5. DOI: 10.1016/j.exppara.2011.06.002CrossRefGoogle Scholar
  10. Gallego, C., Saavedra-Matiz, C., Gomez-Marin J.E. 2006. Direct genotyping of animal and human isolates of Toxoplasma gondii from Colombia (South America). Acta Tropica, 97, 161–7. DOI: 10.1016/j.actatropica.2005.10.001CrossRefGoogle Scholar
  11. Gao, J.M., Xie, Y.T., Xu, Z.S., Chen, H., Hide, G., Yang, T.B., et al. 2017. Genetic analyses of Chinese isolates of Toxoplasma gondii reveal a new genotype with high virulence to murine hosts. Veterinary Parasitology, 241, 52–60. DOI: 10.1016/j.vetpar.2017.05.007CrossRefGoogle Scholar
  12. Howe, D.K., Sibley L.D. 1995. Toxoplasma gondii comprises three clonal lineages: correlation of parasite genotype with human disease. Journal of Infectious Diseases, 172, 1561–6.CrossRefGoogle Scholar
  13. Howe, D.K., Summers, B.C., Sibley L.D. 1996. Acute virulence in mice is associated with markers on chromosome VIII in Toxoplasma gondii. Infection and Immunity, 64, 5193–8PubMedPubMedCentralGoogle Scholar
  14. Lecordier, L., Moleon-Borodowsky, I., Dubremetz, J.F., Tourvieille, B., Mercier, C., Deslee, D., et al. 1995. Characterization of a dense granule antigen of Toxoplasma gondii (GRA6) associated to the network of the parasitophorous vacuole. Molecular and Biochemical Parasitology, 70, 85–94CrossRefGoogle Scholar
  15. Lehmann, T., Marcet, P.L., Graham, D.H., Dahl, E.R., Dubey J.P. 2006. Globalization and the population structure of Toxoplasma gondii. Proceedings of the National Academy of Sciences of the United States of America, 103, 11423–8. DOI: 10.1073/pnas.0601438103CrossRefGoogle Scholar
  16. Lai, M.Y., Lau Y.L. 2018. Measurement of binding strength between prey proteins interacting with Toxoplasma gondii SAG1 and SAG2 using isothermal titration calorimetry (ITC). Acta Parasitologica. 63(1), 106–113. DOI: 10.1515/ap-2018-0012CrossRefGoogle Scholar
  17. Li, K., Wang, M., Zhang, H., Lei, Z., Zhang, L., Luo, H., et al. 2017. Epidemiology of Toxoplasma gondii infection in native tibetans in Tibet, China. Acta Parasitologica, 62(3), 529. DOI: 10.1515/ap-2017-0063CrossRefGoogle Scholar
  18. Li, M., Mo, X.W., Wang, L., Chen, H., Luo, Q.L., Wen, H.Q., et al. 2014. Phylogeny and virulence divergency analyses of Toxoplasma gondii isolates from China. Parasites & Vectors, 7, 133. DOI: 10.1186/1756-3305-7-133CrossRefGoogle Scholar
  19. Li, Y.N., Nie, X., Peng, Q.Y., Mu, X.Q., Zhang, M., Tian, M.Y., et al. 2015. Seroprevalence and genotype of Toxoplasma gondii in pigs, dogs and cats from Guizhou province, Southwest China. Parasites & Vectors, 8, 214. DOI: 10.1186/s13071-015-0809-2CrossRefGoogle Scholar
  20. Qian, W., Wang, H., Su, C., Shan, D., Cui, X., Yang, N., et al. 2012. Isolation and characterization of Toxoplasma gondii strains from stray cats revealed a single genotype in Beijing, China. Veterinary Parasitology, 187, 408–13. DOI: 10.1016/j.vetpar.2012.01.026CrossRefGoogle Scholar
  21. Richomme, C., Aubert, D., Gilot-Fromont, E., Ajzenberg, D., Mercier, A., Ducrot, C., et al. 2009. Genetic characterization of Toxoplasma gondii from wild boar (Sus scrofa) in France. Veterinary Parasitology, 164, 296–300. DOI: 10.1016/j.vetpar.2009.06.014CrossRefGoogle Scholar
  22. Rinder, H., Thomschke, A., Dardé M., Löscher T. 1995. Specific DNA polymorphisms discriminate between virulence and non-virulence to mice in nine Toxoplasma gondii strains. Molecular and Biochemical Parasitology, 69, 123–6CrossRefGoogle Scholar
  23. Saeij, J.P., Boyle, J.P., Coller, S., Taylor, S., Sibley, L.D., Brooke-Powell E.T., et al. 2006. Polymorphic secreted kinases are key virulence factors in toxoplasmosis. Science, 314, 1780–3. DOI: 10.1126/science.1133690CrossRefGoogle Scholar
  24. Sibley, L.D., Ajioka J.W. 2008. Population structure of Toxoplasma gondii: clonal expansion driven by infrequent recombination and selective sweeps. Annual Review of Microbiology, 62, 329–51. DOI: 10.1146/annurev.micro.62.081307.162925CrossRefGoogle Scholar
  25. Su, C., Shwab, E.K., Zhou, P., Zhu, X.Q., Dubey J.P. 2010. Moving towards an integrated approach to molecular detection and identification of Toxoplasma gondii. Parasitology, 137, 1–11. DOI: 10.1017/S0031182009991065CrossRefGoogle Scholar
  26. Su, C., Zhang, X., Dubey J.P. 2006. Genotyping of Toxoplasma gondii by multilocus PCR-RFLP markers: a high resolution and simple method for identification of parasites. International Journal for Parasitology, 36, 841–8. DOI: 10.1016/j.ijpara.2006.03.003CrossRefGoogle Scholar
  27. Wang, D., Liu, Y., Jiang, T., Zhang, G., Yuan, G., He, J., et al. 2016. Seroprevalence and genotypes of Toxoplasma gondii isolated from pigs intended for human consumption in Liaoning province, northeastern China. Parasites & Vectors, 9, 248. DOI: 10.1186/s13071-016-1525-2CrossRefGoogle Scholar
  28. Wang, L., Chen, H., Liu, D.H., Huo, X.X., Gao, J.M., Song, X.R., et al. 2013. Genotypes and mouse virulence of Toxoplasma gondii isolates from animals and humans in China. Plos One, 8, e53483. DOI: 10.1371/journal.pone.0053483CrossRefGoogle Scholar
  29. Windeck, T., Gross U. 1996. Toxoplasma gondii strain-specific transcript levels of SAG1 and their association with virulence. Parasitology Research, 82, 715–9CrossRefGoogle Scholar
  30. Wu, L., Chen, S.X., Jiang, X.G., Fu, X.L., Shen, Y.J., Cao J.P. 2012. Separation and purification of Toxoplasma gondii tachyzoites from in vitro and in vivo culture systems. Experimental Parasitology, 130, 91–4. DOI: 10.1016/j.exppara.2011.10.006CrossRefGoogle Scholar
  31. Xie, D., Weng, Y., Li, H., Zhang, H., Lin, R., Zhang, D., et al. 2005. Comparative studies on GRA6 gene of Toxoplasma gondii isolates from China. Scientia Agricultura Sinica, 38, 1495–500Google Scholar
  32. Yan, C., Liang, L.J., Zhang, B.B., Lou, Z.L., Zhang, H.F., Shen, X., et al. 2014. Prevalence and genotyping of Toxoplasma gondii in naturally-infected synanthropic rats (Rattus norvegicus) and mice (Mus musculus) in eastern China. Parasites & Vectors, 7, 591. DOI: 10.1186/s13071-014-0591-6CrossRefGoogle Scholar
  33. Yang, Y.R., Feng, Y.J., Lu, Y.Y., Dong, H., Li, T.Y., Jiang, Y.B., et al. 2017. Antibody detection, isolation, genotyping, and virulence of Toxoplasma gondii in captive felids from China. Frontiers in Microbiology. 8, 1414. DOI: 10.3389/fmicb.2017.01414CrossRefGoogle Scholar
  34. Zhang, W., Liu, J.S., Ma, Y.W., Lu S. 1987. Toxoplasma gondii isolated from a dead deformed fetus. Chinese Journal of Zoonoses, 3, 26Google Scholar
  35. Zhou, J., Lu, G., Wang, L., Zhou, A.H., Han, Y.L., Guo, J.J., et al. 2017. Structuraland antigenic analysis of a new rhoptry pseudokinase gene (rop54) in Toxoplasma gondii. Acta Parasitologica, 62(3), 513–19. DOI: 10.1515/ap-2017-0061CrossRefGoogle Scholar
  36. Zhu, C., Cheng, W., Luo, Q., Shen J. 2016. Genotypes and virulence of Toxoplasma gondii isolates from cats in Xuzhou. Acta Universitatis Medicinalis Anhui. 10, 1421–5. DOI:10.19405/j.cnki.issn1000-1492.2016.10.006Google Scholar

Copyright information

© Witold Stefański Institute of Parasitology, Polish Academy of Sciences 2018

Authors and Affiliations

  • Zhaofeng Hou
    • 1
    • 2
    • 3
  • Yonghua Zhou
    • 4
  • Dandan Liu
    • 1
    • 2
    • 3
  • Shijie Su
    • 1
    • 2
    • 3
  • Zhenxing Zhao
    • 1
    • 2
    • 3
  • Jinjun Xu
    • 1
    • 2
    • 3
  • Jianping Tao
    • 1
    • 2
    • 3
    Email author
  1. 1.College of Veterinary MedicineYangzhou UniversityYangzhouPeople’s Republic of China
  2. 2.Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhou UniversityYangzhouPeople’s Republic of China
  3. 3.Jiangsu Key Laboratory of ZoonosisYangzhou UniversityYangzhouPeople’s Republic of China
  4. 4.Jiangsu Institute of Parasitic DiseasesWuxiPeople’s Republic of China

Personalised recommendations