Molecular evidence ofand closely relatedspp. in hard ticks infesting domestic animals.

Abstract

Ticks pose significant threats to hosts by transmittingspp., which are grouped into Lyme borreliae, relapsing fever borreliae (RF), and reptiles- and monotremes-associated borreliae. The RF borreliae encompass a group ofspecies predominantly transmitted by soft ticks, but some of its members can also be transmitted by hard ticks. Information on the detection and genetic characterization of tick-borne RF borreliae, including, is notably rare in Asia, particularly in Pakistan. Herein, we employed molecular techniques to detect borreliae in hard ticks collected from domestic animals in Khyber Pakhtunkhwa, Pakistan. Ticks were subjected to morphological analysis, followed by DNA extraction and PCR amplification of partial fragments of borrelial 16S rRNA andgenes. A total of 729 ticks were collected from 264 hosts, with(12.9%; 94/729) being the most prevalent, followed by(11.7%; 85/729),(10.0%; 73/729),(9.1%; 66/729),(8.5%; 62/729),(8%; 58/729),(6.2%; 45/729),(5.5%; 40/729),and(4.4%; 32/729 each),(4.1%; 30/729),and(3.8%; 28/729 each),(2.9%; 21/729),(2.6%; 19/729), and(2.2%; 16/729). Based on 16S rRNA detection ofspp., onlyyielded positive results, resulting in an overall infection rate of 0.3% (2/160), while using-based detection, four tick species including,,, andshowed positive results, yielding an overall infection rate of 6.9% (11/160). The amplified DNA fragments of borrelial 16S rRNA andinfrom goats shared maximum identities of 100 and 99.40% with, respectively. Amplified borrelialfragments infrom cows and sheep displayed 100% identity with, whilefragments inandfrom goats revealed identities of 99.32 and 99.75% with undetermined RFspp., respectively. Phylogenetic analysis revealed clustering offromandwith the same species, whilespp. fromandwith undetermined RFspp. Notably, this research marks the first documentation ofinand the identification of RFspp. inand.