Phylogenetic Position of Haemaphysalis kashmirensis and Haemaphysalis cornupunctata , with Notes on Rickettsia spp

Despite high diversity in the Oriental region, ticks of the genus have been neglected regarding their genetic data and vector potential. This study aimed to genetically characterize three species of the genus : , and infesting goats and sheep, and spp. associated with these tick species in the Hindu Kush Himalayan range of Pakistan. Altogether, 834 ticks were collected by examining 120 hosts including goats (64/120, 53.3%) and sheep (56/120, 46.6%), in which 86 (71.6%) hosts were found to be tick-infested. The morphologically identified ticks were subjected to DNA extraction and PCR for the amplification of partial 16S rDNA and fragments. spp. associated with the collected ticks were detected through the amplification of , and partial fragments. The 16S rDNA of and showed a maximum identity of 100% with the sequences of the same species, whereas the 16S rDNA of showed the highest identity of 93-95% with . The sequence of displayed 100% identity with the same species. In comparison, the sequences of and showed maximum identities of 87.65-89.22% with and 89.34% with , respectively. The sequence of sp. from showed the highest identity of 97.89% with subsp. , while the and fragments from the same DNA samples revealed 100% and 98.16% identity with sp. and " Rickettsia longicornii", respectively. Another sequence amplified from ticks showed 100% identity with , while the attempts to amplify and for were unsuccessful. In the phylogenetic tree, the 16S rDNA of clustered with the corresponding species while its clustered with . Both 16S rDNA and sequences of clustered with . The sequence of sp. was clustered individually in the spotted fever (SF) group of , while the sequence of was clustered with the same species in the transition group of . In the SF group, the rickettsial and sequence clustered with undetermined sp. and " Rickettsia longicornii", respectively. This is the earliest study regarding the genetic characterization of . This study indicated that ticks belong to the genus have the potential of harboring and/or transmitting spp. in the region.