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Molecular and virulence characterization of Toxoplasma gondii strains isolated from humans in Portugal

Parasitology Research volume 116, pages 979–985 (2017)Cite this article

Abstract

Toxoplasma gondii is an apicomplexan parasite responsible for toxoplasmosis which infects all warm-blooded vertebrates, including mammals and birds. The majority of studies conducted in Europe have revealed that more than 80 % of strains isolated from human infections belong to genotype II, whereas genotypes I and III are responsible for a small number of cases. Atypical and recombinant strains are generally associated with more severe infections. In Portugal, there is a lack of data concerning genetic diversity as the classical typing studies in humans have never been performed. We aimed to determine the Sag2 and microsatellite-based (TUB2, TgM-A, W35, B17, B18) genotypes of T. gondii isolated from humans in Portugal, as well as to study their virulence in mice. We analyzed 48 strains from congenital and acquired toxoplasmosis collected during the last two decades. Sag2-based genotyping of T. gondii was achieved in all 48 strains where 35 (73 %) were classified as type II and 13 (27 %) were type I. The multilocus PCR of five microsatellites allowed the classification of 10 strains (21 %) as recombinant strains that had been previously identified as type II or I by Sag2 typing. Seven out of the 48 strains, including three type I, three recombinant, and one type I, were virulent in mice. This study constitutes the first evidence of recombinant strains circulating in Portugal in humans from congenital infection, highlighting the need for a better evaluation of these strains as their phenotype is still barely understood.

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Acknowledgements

We thank Daniel Ajzenberg and Marie-Laure Dardé for all the support knowledge and patience. We thank Assunção António, Paula Grilo, for animal housekeeping, Luis Vieira, Sónia Pedro, Ana Cardoso, and Catarina Silva for their sequencing work.

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Affiliations

  1. National Reference Laboratory of Parasitic and Fungal Infections, Department of Infectious Diseases, National Institute of Health Dr. Ricardo Jorge, Av. Padre Cruz, 1649-016, Lisbon, Portugal

    Anabela Vilares, Maria João Gargaté, Idalina Ferreira & Susana Martins

  2. Research and Development Unit, Infectious Diseases Department, National Institute of Health Dr. Ricardo Jorge, Av. Padre Cruz, 1649-016, Lisbon, Portugal

    João Paulo Gomes

  3. Bioinformatics Unit, Infectious Diseases Department, National Institute of Health Dr. Ricardo Jorge, Av. Padre Cruz, 1649-016, Lisbon, Portugal

    João Paulo Gomes

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  1. Anabela Vilares
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  2. Maria João Gargaté
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  3. Idalina Ferreira
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  4. Susana Martins
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  5. João Paulo Gomes
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Correspondence to Anabela Vilares.

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Funding

Anabela Vilares (SFRH/BD/75154/2010) was the recipient of a PhD fellowship supported by Fundação para a Ciência e Tecnologia.

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The authors declare that they are no conflict of interest.

Electronic supplementary material

Annex 1

Clinical characteristics of patients with Toxoplasma gondii infection, and relationships with the mice virulence. *Strains were numbered solely for simplification purposes considering their complex lab designation. “AF - amniotic fluid, UCB - Umbilical cordon blood, PL – Placenta, BB – Baby blood. #No-Without symptoms; ?-No information available. »NV-Non virulent; V-Virulent.α+Newborn with T. gondii; −Newborn without T. gondii; ?No information available. (DOCX 26 kb).

Annex II

Genotyping analysis REC - Recombinant (DOCX 34 kb).

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Vilares, A., Gargaté, M.J., Ferreira, I. et al. Molecular and virulence characterization of Toxoplasma gondii strains isolated from humans in Portugal. Parasitol Res 116, 979–985 (2017). https://doi.org/10.1007/s00436-017-5374-5

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  • DOI: https://doi.org/10.1007/s00436-017-5374-5

Keywords

  • Toxoplasma gondii
  • Genotype
  • Recombinant
  • Strain
  • Congenital