Abstract
Intra-leukocytic gamonts consistent with the description of Hepatozoon griseisciuri Clark, 1958 are reported for the first time in Canadian eastern gray squirrels (Sciurus carolinensis Gmelin, 1788). Polymerase chain reaction (PCR) amplification and direct Sanger sequencing identified a pair of distinct genotypes at both a nuclear and mitochondrial locus; two 18S ribosomal RNA gene sequences (rDNA; genotype A and genotype B: 1816 base pairs (bp); 98.8% pairwise identity) and 2 distinct complete mitochondrial genome sequences (genotype A: 6311 bp; genotype B: 6114 bp; 89.1% pairwise identity) were obtained from 3 H. griseisciuri-infected squirrels sampled in Guelph, Ontario. The genetic content of both circular-mapping mitochondrial genomes was conventional for apicomplexan protists; each encoded for 3 protein-coding genes (cytochrome c oxidase subunit I (COI); cytochrome c oxidase subunit III (COIII); and cytochrome B (CytB)), 14 fragmented large subunit rDNA, 10 fragmented small subunit rDNA, and 8 unassigned rDNA. These genotypes, based on sequences obtained from a pair of loci from two parasite genomes, confirm the presence of at least two Hepatozoon species infecting Ontario eastern gray squirrels, one of which is likely to be conspecific with H. griseisciuri.
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Acknowledgments
A.N.L. was supported by a Queen Elizabeth II Graduate Scholarship in Science and Technology and an Ontario Graduate Scholarship from the Government of Ontario, and an OVC Scholarship from the Ontario Veterinary College, University of Guelph. Dr. Darren Wood (Pathobiology, University of Guelph) is thanked for his assistance in the specific identification of parasitized squirrel leukocytes. Dr. Todd Smith, Perryn Kruth, Rachel Imai, and Jessica Rotolo are thanked for their comments on earlier drafts of this paper; Perryn Kruth is also thanked for her assistance with squirrel necropsies. The staff of the Guelph Molecular Supercenter, Laboratory Services Division, University of Guelph, are thanked for their diligent hard work.
Funding
This research was supported by a Discovery Grant to J.R.B. from the Natural Sciences and Engineering Research Council of Canada (NSERC-DG #400566).
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Léveillé, A.N., El Skhawy, N. & Barta, J.R. Multilocus sequencing of Hepatozoon cf. griseisciuri infections in Ontario eastern gray squirrels (Sciurus carolinensis) uncovers two genotypically distinct sympatric parasite species. Parasitol Res 119, 713–724 (2020). https://doi.org/10.1007/s00436-019-06583-5
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DOI: https://doi.org/10.1007/s00436-019-06583-5