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Veterinary research communications2024; 49(1); 16; doi: 10.1007/s11259-024-10599-3

Genetic diversity and phylogenetic relationships of Theileria equi inferred from heat shock protein 70 kDa gene sequences in Brazilian horses.

Abstract: Equine piroplasmosis, a tick-borne disease caused by hemoparasites of the Babesia and Theileria genera, has significant clinical and economic impacts worldwide. This study aims to characterize the heat shock protein 70 kDa (hsp70) gene sequences of Theileria equi from naturally infected horses across the five geographic regions in Brazil, and to analyze the phylogenetic relationships between T. equi and other parasites within the order Piroplasmida. Primers specific to T. equi were designed through in silico analysis of hsp70 gene sequences available in GenBank. Forty samples of equine whole blood were subjected to DNA extraction and conventional PCR targeting the hsp70 gene. The resulting amplicons were purified and sequenced. The Hsp70 sequences generated were aligned using the ClustalW method, and phylogenetic analyses were conducted using Bayesian Inference (BI) and Maximum Likelihood (ML) methods. The evolutionary distance within and between species of parasites from the order Piroplasmida was evaluated, as was the entropy of the hsp70 gene sequence among Theileria species. Phylogenetic analysis of the forty sequences identified in this study revealed two circulating genotypes of T. equi in Brazil. One genotype comprised the American sequence along with thirty-six Brazilian sequences, while the other grouped a sequence from Mongolia with four Brazilian sequences. The Equus clade was distinctly placed with strong support, indicating a different evolutionary ancestor compared to Theileria spp. and Babesia spp. groups. The results confirmed hsp70 as an effective molecular marker for phylogeny, yielding topologies similar to those observed with 18 S rDNA while distinguishing closely related protozoan species such as T. haneyi and T. equi. This study provides valuable evolutionary insights into the phylogenetic relationships within the order Piroplasmida.
© 2024. The Author(s), under exclusive licence to Springer Nature B.V.
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Publication Date: 2024-11-19 PubMed ID: 39560826PubMed Central: 5967553DOI: 10.1007/s11259-024-10599-3Google Scholar: Lookup
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Summary

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The research article dissects the genetic diversity and evolutionary relationships of Theileria equi, a parasite affecting horses, using the heat shock protein 70 kDa (hsp70) gene sequences traced from horses in Brazil. This study centers on a disease called equine piroplasmosis, which is caused by tick-borne parasites. The article discusses a detailed analysis of the hsp70 gene’s variation and the genetic relationship between T. equi and other similar parasites.

Research Process

  • The researchers used the hsp70 gene sequences of T. equi, obtained through an in silico analysis, to design specific primers for the said parasite.
  • Using DNA extracted from 40 equine blood samples, a conventional Polymerase Chain Reaction (PCR) was carried out, targeting the hsp70 gene.
  • Theamplicons subsequently obtained were purified and sequenced.
  • The newly generated Hsp70 sequences were placed in proper alignment using the ClustalW method.
  • Phylogenetic relationships were then assessed by adopting the Bayesian Inference (BI) and Maximum Likelihood (ML) methods.
  • The genetic differences within and between species of parasites from the Piroplasmida order were evaluated, as was the hsp70 gene sequence’s difference across Theileria species.

Key findings

  • The phylogenetic analysis of the obtained 40 sequences divulged two circulating genotypes of T. equi in Brazil. 36 Brazilian sequences clustered with the American sequence representing one genotype while the other genotype grouped a Mongolia sequence with four Brazilian sequences.
  • The study established the Equus clade independently with strong backing. This finding indicates a divergent evolutionary origin compared to groups of Theileria spp. and Babesia spp.
  • The results corroborated the hsp70 gene as an efficient molecular marker for phylogeny. It yielded similar topologies to those observed with the 18S rDNA, while distinguishing between closely related protozoan species such as T. haneyi and T. equi.
  • The study provides significant evolutionary insights into the phylogenetic relationships within the Piroplasmida order, furthering our understanding of the genetic intricacies of this group of parasites.

Cite This Article

APA
Paulino PG, Amaral FB, de Oliveira RT, de Andrade SG, Rabello CA, Meirelles N, de Souza Santana M, Galdino KCP, Jacob JCF, Peckle M, Massard CL, Santos HA. (2024). Genetic diversity and phylogenetic relationships of Theileria equi inferred from heat shock protein 70 kDa gene sequences in Brazilian horses. Vet Res Commun, 49(1), 16. https://doi.org/10.1007/s11259-024-10599-3

Publication

Veterinary research communications
ISSN: 1573-7446
NlmUniqueID: 8100520
Country: Switzerland
Language: English
Volume: 49
Issue: 1
Pages: 16

Researcher Affiliations

Paulino, Patrícia Gonzaga
  • Department of Epidemiology and Public Health, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil.
Amaral, Fernanda Barcelos
  • Department of Epidemiology and Public Health, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil.
de Oliveira, Roberto Teixeira
  • Department of Epidemiology and Public Health, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil.
de Andrade, Sara Gomes
  • Department of Epidemiology and Public Health, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil.
Rabello, Carla Alves
  • Department of Epidemiology and Public Health, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil.
Meirelles, Nelson
  • Department of Epidemiology and Public Health, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil.
de Souza Santana, Matheus
  • Department of Epidemiology and Public Health, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil.
Galdino, Karina Cristina Paes
  • Department of Epidemiology and Public Health, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil.
Jacob, Júlio César Ferraz
  • Department of Animal Reproduction and Evaluation, Federal Rural University of Rio de Janeiro (UFRRJ), Seropedica, RJ, Brazil.
Peckle, Maristela
  • Department of Animal Parasitology, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil.
Massard, Carlos Luiz
  • Department of Animal Parasitology, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil.
Santos, Huarrisson Azevedo
  • Department of Epidemiology and Public Health, Federal Rural University of Rio de Janeiro (UFRRJ), BR 465, Km 7, Seropedica, 23890000, RJ, Brazil. huarrisson@yahoo.com.br.
  • Department of Epidemiology and Public Health, Veterinary Institute, Federal Rural University of Rio de Janeiro, BR 465, Km 7, Rio de Janeiro, Seropedica, 23890-000, Brazil. huarrisson@yahoo.com.br.

MeSH Terms

  • Animals
  • Theileria / genetics
  • Theileria / classification
  • Horses
  • HSP70 Heat-Shock Proteins / genetics
  • Phylogeny
  • Horse Diseases / parasitology
  • Brazil
  • Theileriasis / parasitology
  • Genetic Variation

Grant Funding

  • E-26/201.342/2021 / Fundau00e7u00e3o Carlos Chagas Filho de Amparo u00e0 Pesquisa do Estado do Rio de Janeiro
  • 313753/2021-0, 403524/2023-7 / Conselho Nacional de Desenvolvimento Cientu00edfico e Tecnolu00f3gico

Conflict of Interest Statement

These procedures were approved by the Ethics Committee on Research of the Federal Rural University of Rio de Janeiro, process number 2014/0003 (Date July 13th, 2015). The authors declare no competing interests.

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Citations

This article has been cited 1 times.
  1. Braga ÍA, de Urzedo NF, Maia GO, Gomes APC, Duarte RB, Alves-Ribeiro BS, da Silva Martins A, Silva TA, Pires LHB, Rezende SR, de Assis Pereira N, Paludo RLDR, Regalin D, Rezende HHA, Saturnino KC, de Souza Ramos DG, de Aguiar DM, Borsanelli AC. Feline botulism: two decades and two reports. Vet Res Commun 2025 Aug 9;49(5):279.
    doi: 10.1007/s11259-025-10848-zpubmed: 40782285google scholar: lookup
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