First report of Hepatozoon and Lankesterella spp. infections in wild rodents from Pakistan, and their potential impact on blood parameters and oxidative stress markers in vital organs.

Pakistan is home to a rich rodent fauna, yet no investigations have been conducted on the presence of Haemogregarines in these animals, leaving their epidemiology and genetic diversity in this subtropical region unexplored. In this study, blood samples from four wild rodent species, Rattus (R.) rattus (n = 122), Mus (M.) musculus (n = 64), Rattus norvegicus (n = 57), and Dryomys nitedula (n = 1), were collected between May 2022 and July 2023 from three districts in Punjab and three in Khyber Pakhtunkhwa. PCR targeting a fragment of 600 base pairs from the 18 S rDNA gene revealed a 2.86% (7/244) prevalence of Haemogregarines in the rodents. DNA sequencing and BLAST analysis confirmed the presence of Hepatozoon spp. and Lankesterella spp. in the blood samples. Phylogenetic analysis indicated genetic diversity in the Pakistani sequences, which clustered with sequences found in reptiles, amphibians, birds, and mammals from various countries. The prevalence of the parasite varied among rodent species, with R. rattus showing the highest infection rate, followed by R. norvegicus and M. musculus. Female rodents were more frequently infected than males. Infected R. rattus exhibited significant disruptions in white blood cells, red blood cells, and platelet counts. Oxidative stress markers indicated elevated superoxide dismutase in the kidney, catalase in the heart, and malondialdehyde in the liver and lungs of infected rodents compared to uninfected ones. This study is an important contribution towards science as it the first report of Haemogregarines and Lankesterella spp. infections among Pakistani rodents.