Molecular Identification and Phylogenetic Analysis of <em>Trypanosoma evansi</em> with Assessment of Associated Risk Factors in Camels (<em>Camelus dromedarius</em>) Across Ten Districts of Punjab, Pakistan

Trypanosomiasis significantly impacts camel health and productivity, posing a major challenge to food security in regions with large camel populations. This study investigat-ed the microscopic as well as molecular prevalence, phylogenetic analysis, and risk fac-tors associated with Trypanosoma evansi (T. evansi) infection in 400 randomly selected suspected camels (Camelus dromedarius) from 10 districts of Punjab, Pakistan. Blood samples were to microscopic examination of Giemsa / Field’s-stained smears, and a three PCR primer sets (ITS1CF/BR, pMUTec, RoTat 1.2) to detect the presence of T. evansi. PCR-based prevalence was higher (14.8%; CI 11.4-18.6) as compared to the microscopic examination (8.3%; CI 5.7-11.4) of samples. The targeted primers amplified DNA frag-ments of 210, 205, and 478 base pairs, respectively. Phylogenetic analysis showed 100% homology between local isolates and those from India, Sudan, Malaysia, Egypt, and Ken-ya. Risk analysis identified female gender (OR 2.1) and being in Southern Punjab (OR: 1.9) as significant factors associated with disease. Significantly (p &lt; 0.05) reduced total protein (5.51 + 0.05), albumin (2.77 + 0.04), and globulin (2.57 + 0.06) levels were found in PCR positive camels. This study provides new molecular and phylogenetic data on T. evansi in Pakistan.