Atypical Toxoplasma gondii genotype from a sheep and a pig on Fernando de Noronha Island, Brazil, showed different mouse virulence profiles

  • Renata Pimentel B. MeloEmail author
  • Jonatas C. Almeida
  • Débora C. V. de Lima
  • Jéssica C. S. Carvalho
  • Wagnner J. N. Porto
  • Fernando J. R. Magalhães
  • Clare M. Hamilton
  • Frank Katzer
  • Rinaldo A. Mota
Protozoology - Short Communication


Toxoplasma gondii is a zoonotic parasite which can infect almost all warm-blooded animals. Toxoplasma gondii isolates from Brazil have greater genetic diversity with a predominance of virulent and atypical genotypes, compared with the Northern Hemisphere. Considering that previous studies have demonstrated a high seroprevalence of T. gondii antibodies in animals from Fernando de Noronha Island, the aim of this study was to isolate, genetically characterize, and determine mouse virulence of isolates of T. gondii from livestock from this Brazilian island. Two T. gondii isolates were obtained by mouse bioassay from brain from one sheep and one pig. Genotyping was performed by PCR-RFLP using 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22- 8, c29-2, PK1, L358, and Apico) and an atypical genotype of T. gondii (ToxoDB #146) was identified for both isolates. Genotyping of four ROP loci indicated different alleles for ROP16 and mouse virulence analysis revealed different profiles (intermediate and low virulence). This is the first report of this genotype being described in a pig and a sheep.


Mouse bioassay Isolation Genotyping Virulence genes 



The authors thank the Health Superintendence of Fernando de Noronha Island for all the support during the sample collection.

Funding information

This study was supported by Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco (FACEPE, APQ-0531-5.05/14). We would like to thank to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Brazil, for the scholarship (SWE, grant number 206740/2017-4) and to FACEPE for the PhD scholarship (IBPG-1358-5.05./15). The authors thank the Health Superintendence of Fernando de Noronha Island for all the support during the sample collection.

Compliance with ethical standards

All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. The study was approved by the Ethics Committee in Animal Use at Universidade Federal Rural de Pernambuco (UFRPE) (license number 139/2016).

Conflict of interest

The authors declare that they have no conflicts of interest.


  1. Almeida JC, Melo RPB, Pedrosa CM, Santos MS, Barros LD, Garcia JL, Porto WJN, Mota RA (2017) First isolation and RFLP genotyping of Toxoplasma gondii from crab-eating fox (Cerdocyon thous - Linnaeus, 1766). Acta Trop 169:26–29. CrossRefPubMedGoogle Scholar
  2. Burrells A, Bartley PM, Zimmer IA, Roy S, Kitchener AC, Meredith A, Wright SE, Innes EA, Katzer F (2013) Evidence of the three main clonal Toxoplasma gondii lineages from wild mammalian carnivores in the UK. Parasitol 140:1768–1776. CrossRefGoogle Scholar
  3. Camargo ME (1964) Improved technique of indirect immunofluorescence for serological diagnosis of toxoplasmosis. Rev Inst Med Trop Sao Paulo 6:117–118PubMedGoogle Scholar
  4. Cenci-Goga BT, Rossitto PV, Sechi P, McCrindle CM, Cullor JS (2011) Toxoplasma in animals, food, and humans: an old parasite of new concern. Foodborne Pathog Dis 8(7):751–762. CrossRefPubMedGoogle Scholar
  5. Dubey JP (1998) Refinement of pepsin digestion method for isolation of Toxoplasma gondii from infected tissues. Vet Parasitol 74:75-77. Scholar
  6. Dubey JP (2010) Toxoplasmosis of animals and humans, 2nd edn. CRC Press, Boca Raton, FloridaGoogle Scholar
  7. Dubey JP, Rajendran C, Costa DGC, Ferreira LR, Kwok OCH, Qu D, Su C, Marvulo MFV, Alves LC, Mota RA, Silva JCR (2010) New Toxoplasma gondii genotypes isolated from free-range chickens from the Fernando de Noronha, Brazil: Unexpected findings. J Parasitol 96(4):709–712. CrossRefPubMedGoogle Scholar
  8. Etheridge RD, Alaganan A, Tang K, Lou HJ, Turk BE, Sibley LD (2014) The Toxoplasma pseudokinase ROP5 forms complexes with ROP18 and ROP17 kinases that synergize to control acute virulence in mice. Cell Host Microbe 15:537–550. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Garcia JL, Navarro IT, Ogawa L, De Oliveira RC (1999) Seroprevalence of Toxoplasma gondii in pigs, cattle, sheeps and equines and its correlation with humans, felines and canines from rural areas in north region of Paraná State. Brazil Cien Rural 29:91–97. CrossRefGoogle Scholar
  10. Hamilton CM, Kelly PJ, Bartley PM, Burrells A, Porco A, Metzler D, Crouch K, Ketzi JK, Innes EA, Katzer F (2015) Toxoplasma gondii in livestock in St. Kitts and Nevis. West Indies Parasit Vectors 8(166):1–9. CrossRefGoogle Scholar
  11. Hamilton CM, Black L, Oliveira S, Burrells A, Bartley PM, Melo RPB, Chianini F, Palarea-Albaladejo J, Innes EA, Kelly PJ, Katzer F (2019) Comparative virulence of Caribbean, Brazilian and European isolates of Toxoplasma gondii. Parasit Vectors 12(104):1–9. CrossRefGoogle Scholar
  12. Huson DH, Bryant D (2006) Application of phylogenetic networks in evolutionary studies. Mol Biol Evol 23:254–267. CrossRefPubMedGoogle Scholar
  13. Lehmann T, Marcet PL, Graham DH, Dahl ER, Dubey JP (2006) Globalization and the population structure of Toxoplasma gondii. Proc Natl Acad Sci 103:11423–11428. CrossRefPubMedGoogle Scholar
  14. Li JX, He JJ, Elsheikha HM, Chen D, Zhai BT, Zhu XQ, Yan HK (2019) Toxoplasma gondii ROP17 inhibits the innate immune response of HEK293T cells to promote its survival. Parasitol Res 118(3):1–10. CrossRefGoogle Scholar
  15. Lima DCV, Melo RPB, Almeida J, Magalhães FJR, Ribeiro-Andrade M, Pedrosa CM, CAV O, WJN P, Su C, Mota RA (2019) Toxoplasma gondii in invasive animals on the Island of Fernando de Noronha in Brazil: molecular characterization and mouse virulence studies of new genotypes. Comp Immunol Microbiol Infect Dis 67:101347. CrossRefGoogle Scholar
  16. Magalhães FJR, Silva JG, Andrade MR, Junior JWP, Mota RA (2016a) High prevalence of toxoplasmosis in free-range chicken of the Fernando de Noronha Archipelago, Brazil. Acta Trop 159:58–61. CrossRefPubMedGoogle Scholar
  17. Magalhães FJR, Ribeiro-Andrade M, Alcântara AM, Pinheiro-Júnior WP, Sena MJ, Porto WJN, Vieira RFC, Mota RA (2016b) Risk factors for Toxoplasma gondii infection in sheep and cattle from Fernando de Noronha Island, Brazil. Braz J Vet Parasitol 25(4):511–515. CrossRefGoogle Scholar
  18. Magalhães FJR, Ribeiro-Andrade M, Souza FM, Lima-Filho CDF, Biondo AW, Vidotto O, Navarro IT, Mota RA (2017) Seroprevalence and spatial distribution of Toxoplasma gondii infection in cats, dogs, pigs and equines of the Fernando de Noronha Island, Brasil. Parasitol Int 66:43–46. CrossRefPubMedGoogle Scholar
  19. Melo RPB, Almeida JC, Lima DCV, Pedrosa CM, Magalhães FJR, Alcântara AM, Barros LD, Vieira RFC, Garcia JL, Mota RA (2016) Atypical Toxoplasma gondii genotype in feral cats from the Fernando de Noronha Island, northeastern Brazil. Vet Parasitol 224:92–95CrossRefGoogle Scholar
  20. Pena HFJ, Gennari SM, Dubey JP, Su C (2008) Population structure and mouse virulence of Toxoplasma gondii in Brazilian. Int J Parasitol 38:561–569CrossRefGoogle Scholar
  21. Penambuco, 2015. Governo do Estado. Fauna do Arquipélago de Fernando de Noronha. Available from: http://
  22. Reese ML, Zeiner GM, Saeij JP, Boothroyd JC, Boyle JP (2011) Polymorphic family of injected pseudokinases is paramount in Toxoplasma virulence. Proc Natl Acad Sci U S A 108:9625–9630. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Rêgo WMF, Costa JGL, Baraviera RCA, Pinto LV, Bessa GL, Lopes REN, Vitor RWA (2017) Association of ROP18 and ROP5 was efficient as a marker of virulence in atypical isolates of Toxoplasma gondii obtained from pigs and goats in Piauí, Brazil. Vet Parasitol 247:19–25. CrossRefPubMedGoogle Scholar
  24. Saraf P, Shwab EK, Dubey JP, Su C (2017) On the determination of Toxoplasma gondii virulence in mice. Exp Parasitol 174:25–30. CrossRefPubMedGoogle Scholar
  25. Schulz-Neto A (2004) Aves insulares do arquipélago de Fernando de Noronha. p. 147-168 in Aves marinhas e insulares brasileiras: bioecologia e conservação (Organized by Joaquim Olinto Branco). UNIVALI Company, Itajaí, Santa Catarina.Google Scholar
  26. Shwab EK, Jiang T, Pena HF, Gennari SM, Dubey JP, Su C (2016) The ROP18 and ROP5 gene allele types are highly predictive of virulence in mice across globally distributed strains of Toxoplasma gondii. Int J Parasitol 46:141–146. CrossRefPubMedGoogle Scholar
  27. Silva JCR, Ferreira F, Dias RA, Ajzenberg D, Marvulo MFV, Magalhaes FJR, Lima-Filho CDF, Oliveira S, Soares HS, Feitosa TF, Aizawa J, Alves LC, Mota RA, Dubey JP, Gennari SM, Pena HFJ (2017) Cat-rodent Toxoplasma gondii Type II-variant circulation and limited genetic diversity on the Island of Fernando de Noronha, Brazil. Parasit Vectors 10(220):1–6. CrossRefGoogle Scholar
  28. Su C, Shwab EK, Zhou P, Zhu XQ, Dubey JP (2010) Moving towards an integrated approach to molecular detection and identification of Toxoplasma gondii. Parasitol 137:1–11. CrossRefGoogle Scholar
  29. Taylor S, Barragan A, Su C, Fux B, Fentress SJ, Tang K, Beatty WL, Hajj HE, Jerome M, Behnke MS, White M, Wootton JC, Sibley LD (2006) A secreted serine-threonine kinase determines virulence in the eukaryotic pathogen Toxoplasma gondii. Science 314:1776–1780. CrossRefPubMedGoogle Scholar
  30. Tenter AM, Heckeroth AR, Weiss LM (2000) Toxoplasma gondii: from animals to human. Int J Parasitol 30:1217-1258. Scholar
  31. Vitaliano SN, Soares HS, Minervino AHH, Santos ALQ, Werther K, Marvulo MFV, Siqueira DB, Pena HFJ, Soares RM, Su C, Gennari SM (2014) Genetic characterization of Toxoplasma gondii from Brazilian wildlife revealed abundant new genotypes. Int J Parasitol Parasites Wildl 3:276–283. CrossRefPubMedPubMedCentralGoogle Scholar
  32. Yamamoto M, Standley DM, Takashima S, Saiga H, Okuyama M, Kayama H, Kubo E, Ito H, Takaura M, Matsuda T, Soldati-Favre D, Takeda K (2009) A single polymorphic amino acid on Toxoplasma gondii kinase ROP16 determines the direct and strain-specific activation of Stat3. J Exp Med 206:2747–2760. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Renata Pimentel B. Melo
    • 1
    Email author
  • Jonatas C. Almeida
    • 1
  • Débora C. V. de Lima
    • 1
  • Jéssica C. S. Carvalho
    • 1
  • Wagnner J. N. Porto
    • 2
  • Fernando J. R. Magalhães
    • 3
  • Clare M. Hamilton
    • 4
  • Frank Katzer
    • 4
  • Rinaldo A. Mota
    • 1
  1. 1.Department of Veterinary Medicine, Laboratory of Infectious-Contagious Diseases of Domestic AnimalsUniversidade Federal Rural de PernambucoRecifeBrazil
  2. 2.Biological and Health Science InstituteUniversidade Federal de AlagoasMaceióBrazil
  3. 3.Health SuperintendenceAdministração do Distrito Estadual de Fernando de NoronhaPernambucoBrazil
  4. 4.Moredun Research InstituteScotlandUK

Personalised recommendations