Tehran University of Medical Sciences Iranian Journal of Parasitology 1735-7020 21 01 2026 05 19 94 104 Maryam Saeed University of Veterinary and Animal Sciences, Lahore, Pakistan Dr. Asia Iqbal university of veterinary and animal sciences, Lahore Shahzad Ali University of Veterinary and Animal Sciences, Lahore, Pakistan Haroon Akbar University of Veterinary and Animal Sciences, Lahore, Pakistan Saba Sana University of Veterinary and Animal Sciences, Lahore, Pakistan Usama Saeed University of Copenhagen, Copenhagen, Denmark 2025 08 11 Background: This study investigated the prevalence of T. lewisi in Black rats (R. rattus) and Brown rats (R. norvegicus) using both microscopic and molecular detection methods, along with the analysis of associated risk factors. Methods: A total of 178 rodents were trapped in Kasur district, with epidemiological data and geographical coordinates recorded. Rodents were identified, euthanized and blood samples were collected. T. lewisi was confirmed through microscopy, PCR, sequencing and phylogenetic analysis. In the statistical analysis, descriptive statistics, Chi-square and Binary logistic regression were performed using SPSS Software. A GIS map was generated using ArcGIS 10.5.1 to illustrate geographical distribution. Results: The 178 blood samples. PCR confirmed to be either microscopically positive or negative samples. Overall prevalence of T. lewisi was 10(5.62%). At the species level, T. lewisi prevalence was higher in Rattus rattus (7.59%) compared to R. norvegicus (4.04%). Gender and the presence of ectoparasites were identified as potential risk factors. Conclusion: These findings confirm the presence of T. lewisi in black and brown rats and serve as a baseline for further surveillance and control strategies. https://ijpa.tums.ac.ir/index.php/ijpa/article/view/4728 https://ijpa.tums.ac.ir/index.php/ijpa/article/download/4728/1483