Abstract
The order Piroplasmida encompasses tick-borne pathogens of veterinary and medical importance positioned in two main families: Babesiidae and Theileriidae. Even though previous studies carried out in Brazil recorded the occurrence of piroplasmid species circulating in small mammals, 18S RNA gene sequences were only partially sequenced, preventing the assessment of their phylogenetic positioning. The current study aimed to detect and characterize, using morphological, molecular, and bioinformatic approaches, piroplasmids from wild mammals and associated ticks sampled in Central-Western Brazil. Out of 67 Didelphis albiventris sampled, 22 (16.4%) were positive for piroplasmids by PCR. In contrast, none of the 48 small rodents and 14 capybaras (Hydrochoerus hydrochaeris) was PCR-positive. Four Amblyomma dubitatum ticks—one from Rattus rattus, one from H. hydrochaeris, and two from D. albiventris—out of 114 Amblyomma spp. DNA samples were positive for piroplasmids by PCR. The phylogenetic inference performed using the near-complete 18S rRNA gene positioned the putative novel piroplasmid species detected in D. albiventris and associated A. dubitatum ticks near to Babesia sensu lato clade (Western group—cluster III) and distant from the Australian marsupial-associated piroplasms. Phylogenetic inferences based on two additional molecular markers, namely hsp-70 and cox-1, supported the near-complete 18S rRNA gene phylogenetic inference. Finally, the partial 18S rRNA gene sequences detected in ticks from rodents (R. rattus and H. hydrochaeris) showed 97.2–99.4% identity with the Piroplasmida previously detected in a capybara from Brazil, raising evidence that a still uncharacterized piroplasmid species has been identified in the capybara, the largest rodent species from South America.
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Acknowledgements
This work was financially supported by FAPESP (Foundation for Research Support of the State of São Paulo—process numbers 2018/02753-0 and 2020/12037-0), FUNDECT (Foundation for Support to the Development of Education, Science and Technology of the State of Mato Grosso do Sul, Case 59/300.187/2016), and CNPq (National Council for Scientific and Technological Development) for the Productivity Grant to M. R. A. (CNPq Process #302420/2017-7) and H. M. H. (CNPq Process #308768/2017-5). T. B. B. and M. A. D. received scholarship from CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior). The authors are especially thankful to the InsanaHuna Research Group (www.insanahuna.com) for the fieldwork support and to the reviewers whose suggestions significantly improved the paper.
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S1 Table
Occurrence of Piroplasmida DNA in the marsupials, rats, capybaras and their related ticks through different target genes (XLSX 11 KB)
S2 Table
Pairwise genetic distances matrix among Piroplasmida clusters and those detected in the present study at the large 18S rRNA fragment (~1.3 kb). Pairwise genetic distances (%) were obtained using the p-distance method in MEGA X (XLSX 17 KB)
S3 Table
Pairwise genetic distances matrix among Piroplasmida clusters and those detected in the present study at the hsp-70 fragment (~700 bp). Pairwise genetic distances (%) were obtained using the p-distance method in MEGA X. (XLSX 14 KB)
S4 Table
Pairwise genetic distances matrix among Piroplasmida clusters and those detected in the present study at the cox-1 fragment (~800 bp). Pairwise genetic distances (%) were obtained using the p-distance method in MEGA X. (XLSX 14 KB)
S5 Table
Pairwise genetic distances matrix among Piroplasmida clusters and those detected in the present study at the small 18S rRNA fragment (~750 bp). Pairwise genetic distances (%) were obtained using the p-distance method in MEGA X (XLSX 19 KB)
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Gonçalves, L.R., Paludo, G., Bisol, T.B. et al. Molecular detection of piroplasmids in synanthropic rodents, marsupials, and associated ticks from Brazil, with phylogenetic inference of a putative novel Babesia sp. from white-eared opossum (Didelphis albiventris). Parasitol Res 120, 3537–3546 (2021). https://doi.org/10.1007/s00436-021-07284-8
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DOI: https://doi.org/10.1007/s00436-021-07284-8