Molecular Epidemiology of Tick-Borne (Anaplasma, Babesia, Theileria) and Non-Tick Borne (Toxoplasma gondii) Pathogens in Goats from Southern Punjab, Pakistan.

Pakistan ranks third globally in goat population with approximately 89 million heads in 2025; however, despite this large population base, the apparent PCR based prevalence of hemoparasitic and bacterial pathogens in goats remains largely underexplored. This study aimed to report the molecular prevalence and phylogenetic characteristics of Anaplasma ovis, Anaplasma marginale, Babesia ovis, Theileria ovis and Toxoplasma gondii in goat blood samples (n = 247) collected from two districts (Muzaffargarh and Multan) in Punjab between August till November 2025. Results indicated a significant variation in the infection rates of the five screened pathogens among the enrolled goats (P < 0.001). Anaplasma ovis exhibited the highest apparent PCR based prevalence (20.6%), followed by B. ovis (8.5%), T. gondii (8.1%), A. marginale (6.9%) and T. ovis (5.7%) respectively. Instances of co-infection with two and three pathogens were also recorded. DNA sequencing and BLAST analysis confirmed the presence of all screened pathogens. Phylogenetic analysis showed that these isolates were genetically closely related to strains reported from various countries worldwide. We observed higher PCR detected T. gondii infection in smaller herds (P = 0.02) and in herds with dogs (P = 0.03), among Rajanpuri breed (P = 0.005) and in goats from Muzaffargarh district (P = 0.04). In contrast, T. ovis infection was more common among small goat herds (P = 0.04). In conclusion, this study reports a high apparent prevalence of A. ovis while moderate to low PCR based prevalence of A. marginale, B. ovis, T. ovis and T. gondii infections in goats from Punjab, Pakistan. Comprehensive multi-regional studies are warranted to clarify the epidemiology, genetic diversity, and host-parasite dynamics of these pathogens to support effective control strategies in goats.