First detection and genetic identification of Hepatozoon canis in Rhipicephalus sanguineus sensu lato ticks collected from dogs of Taiwan.

We determined the prevalence of infection and genetic identity of Hepatozoon spp. harbored by Rhipicephalus sanguineus sensu lato ticks in Taiwan. A total of 1082 ticks were collected from dogs and DNA extraction was performed from individual tick specimens. Hepatozoon infection was detected by performing a nested-PCR assay based on the 18S small subunit ribosomal RNA (ssrRNA) gene. The genetic identity of detected Hepatozoon was identified by gene sequencing and phylogenetic analysis. Hepatozoon infection was detected in nymphs, males and females of R. sanguineus s. l. ticks with an infection rate of 20.8%, 22.8% and 29.4%, respectively. Sequence and phylogenetic analysis revealed that these Hepatozoon spp. of Taiwan were genetically affiliated to the same clade within the genospecies of H. canis and can be discriminated from other genospecies of H. americanum and H. felis. Intraspecies analysis based on the genetic distance (GD) values indicates a lower level (GD < 0.005) genetic divergence within the same genospecies of H. canis detected in Taiwan, Brazil and Spain. Interspecies analysis also reveals a higher heterogeneity of Taiwan strains distinguished from other genospecies of H. felis (GD > 0.040) and H. americanum (GD > 0.056). This study provides the first molecular evidence of H. canis detected and identified in various stages of R. sanguineus s. l. ticks in Taiwan. Detection of H. canis in unfed male ticks may imply the possible mechanism of transstadial survival in R. sanguineus s. l. ticks. Further investigations on Hepatozoon spp. harbored by various vector ticks in Taiwan may illustrate the epidemiological significance of this parasite.