First molecular identification of Dermacentor montanus and new locality record of Argas hermanni from Hindu Kush and Himalayan ranges.

Soft ticks of the genus Argas are primarily associated with avian hosts, whereas hard ticks of the genus Dermacentor parasitize mammals. This study provides the morphological and molecular characterization of ticks collected from the nests of wild pigeons (Columba livia) and domestic yak (Bos grunniens) inhabiting Himalayan and Hindu Kush ranges of northern Pakistan, with emphasis on wildlife associated niches. A total of 146 ticks including 40 males, and 63 females and 43 nymphs of Argas hermanni were collected from nests inhabited by wild pigeons in District Abbottabad. Moreover, 91 Dermacentor montanus ticks comprising 38 males and 53 females were reported from domestic yak in District Chitral. Adults and nymphs of A. hermanni have a broadly oval, dorsoventrally flattened body with a rounded posterior margin and no mammillae. The integument shows distinct radiating wrinkles, and legs are inserted anteriorly with coxae IV positioned anterior to midlength. The capitulum is ventral, with short palps and a hypostome bearing 2/2 dentition. A semicircular genital aperture is present in adults, while nymphs show a closed genital depression. Morphological identification of D. montanus was established based on key diagnostic characters, including an ornamented conscutum with dense punctation, well-defined cervical grooves, characteristic coxal spurs, and a 3/3 hypostomal dentition pattern; importantly, it was distinguished from Dermacentor pavlovskyi by the morphology of the spiracular plates, which in D. montanus exhibit a minute dorsal prolongation, appearing nearly rounded, whereas in D. pavlovskyi the spiracular plates possess a distinct and pronounced dorsal projection. BLAST analysis of 16S rDNA and cox1 sequences from A. hermanni showed the highest identity of 98.17-99.69% with the same species reported from Iran. The BLAST results of 16S rDNA and cox1 sequences from D. montanus revealed highest identity of 99.65-99.75% with D. montanus and D. pavlovskyi from China. Phylogenetic analyses based on 16S and cox1 demonstrated that A. hermanni grouped with conspecific sequences, forming a well-supported clade, whereas Dermacentor montanus grouped with D. montanus and D. pavlovskyi, indicating close evolutionary relatedness within the subgenus Asiacentor. These findings represent the morphological and genetic identification of A. hermanni and D. montanus in Pakistan and highlight previously undocumented tick-host associations, specifically the occurrence of Argas in wild pigeon nesting and D. montanus on domestic yak, expanding current knowledge of tick distribution and host ecology.