First report on detection of in and in and : risks of spillover of spp. from wildlife to domestic animals.

This study aimed to detect spp. in ticks infesting asymptomatic domestic animals and to provide insight into their potential spillover from wild to domestic animals. In total, 537 tick specimens were collected in Khyber Pakhtunkhwa, Pakistan, and morphologically identified. The most prevalent tick species was (69; 12.8%), followed by (62; 11.5%), (58; 10.8%), (51; 9.5%), (49; 9.1%), each and (43; 8.0%), each and (37; 6.9%), (33; 6.1%) (30; 5.6%), and (25; 4.7%). The extracted DNA from a subset of each tick species was subjected to PCR to amplify 18S rRNA fragments of spp. By BLAST analysis, the sp. detected in infesting cows and in infesting sheep showed 99.7% maximum identity with . Similarly, the sp. detected in infesting goats shared 99.49% maximum identity with , and the sp. detected in infesting dogs exhibited 99.7% identity with . Having an overall infection rate (9.3%; 16/172), the highest infection rate was recorded for each , and (3.5%; 6/172), followed by (2.3%; 4/172). In the phylogenetic tree, clustered with corresponding species from Iran, clustered with the same species from Croatia, Ghana, and Portugal, and clustered with the conspecifics from Iran, Israel, Romania, and Zambia. Regarding the potential spillover of spp. from wildlife through ticks, free ranging animals was at higher risk compared to confined animals (RR = 3.05), animals consuming food from wildlife habitats were at higher risk compared to those consuming domestic food (RR = 3.06), and animals residing in farm buildings located in wildlife habitats were at higher risk compared to those residing in farm buildings located in villages (RR = 3.28). In addition to the first report on in in Pakistan, this is the earliest data showing in and in and . These preliminary findings suggest a potential spillover of spp. from wild to domestic animals via ticks under certain risk factors.