Meta-transcriptomic characterization reveals viral species with zoonotic potential in Rhipicephalus microplus and Haemaphysalis bispinosa ticks in Pakistan

Tick-borne viruses are an increasing global health concern, but the tick virome in South Asia, particularly in Pakistan, remains poorly underexplored. Pakistan's subtropical climate and agricultural environment provide favorable habitats for ticks, making it an endemic region for various tick-borne pathogens. This study aimed to characterize the tick-associated virome and viral diversity of ticks in Khyber Pakhtunkhwa Province, Pakistan, and assess the potential risks to human and livestock health. A meta-transcriptomic investigation was conducted on host seeking adult ticks collected from livestock at 19 sites across the province. A total of 23 pooled samples, consisting of 165 individual ticks of Rhipicephalus microplus and Haemaphysalis bispinosa, were analyzed. The viral characterization involved diversity assessment and phylogenetic analysis, identifying ten RNA virus species from nine families, including two previously undefined novel viruses, below the ICTV species demarcation thresholds. Notably, Mogiana tick virus has been reported to be pathogenic to humans, whereas Pakistan microplus virus and Uukuvirus lihanense are phylogenetically closely related to viruses infecting humans, suggesting a potential risk of cross-species transmission to both humans and livestock. This study provides the first comprehensive dataset of the tick virome in Pakistan, highlighting the need for enhanced surveillance of tick-borne viral infections in humans and livestock.