Molecular epidemiology and genetic profiling of Anaplasma ovis, Bartonella sp., and Toxoplasma gondii in domestic cats.

Cats are among the most common and cherished domestic animals globally. Being our common companion, they are frequently exposed to the environment and encounter different vectors, that makes them carriers of numerous pathogens. Present study was aimed to determine the PCR based prevalence and phylogenetic analysis of Anaplasma ovis, Bartonella sp. and Toxoplasma gondii in pet cats from Pakistan. A total of 144 cat blood samples were collected from two districts in Khyber Pakhtunkhwa (KPK), namely Lower Dir (n = 74) and Buner (n = 70) during September 2024 till May 2025. Polymerase Chain Reaction (PCR) based analysis revealed that 15%, 7% and 4% cats were infected with A. ovis, T. gondii and Bartonella sp., respectively. Co-infections involving two pathogens were also detected. The presence of all three pathogens was confirmed through Sanger sequencing followed by BLAST analysis and they were similar to sequences deposited from countries worldwide. Prevalence of all three pathogens did not vary between the sampling sites or with the cat sex. Bartonella spp. infection was linked to reduced red cell distribution width-coefficient of variation and platelet counts, indicating possible hematological alterations Overall, our findings indicated moderate infection rates of A. ovis, Bartonella sp. and T. gondii in cats from Pakistan. These findings highlight the need to include local cat populations in future disease surveillance programs across Pakistan, following the One Health approach.