Molecular prevalence of Hepatozoon ophisauri and Toxoplasma gondii in the blood samples of wild lizards collected from various altitudes in Khyber Pakhtunkhwa, Pakistan

Despite of having rich reptilian fauna, the wild Pakistani lizards remained unexplored for the blood borne parasites. Hence, this study was designed to report the molecular prevalence and phylogenetic evaluation of Hepatozoon ophisauri and Toxoplasma gondii in blood samples of wild lizards (N = 101) that were trapped from various areas having different altitudes (ranging between 1200 to 2250 m above sea level) in Khyber Pakhtunkhwa province in Pakistan during March 2022 till June 2023. Molecular studies revealed that 3 out of 101 (03%) lizards were Hepatozoon spp. infected. All infected lizards were Laudakia ( L .) tuberculata. Forty five out of one hundred and one lizards (45%) were positive for the presence of Toxoplasma gondii and infected lizards included L. agrorensis , L. pakistanica , L. tuberculate and Abblepharus (A.) pannonicus. DNA sequencing and BLAST analysis confirmed the presence of Hepatozoon ophisauri and Toxoplasma gondii. Phylogenetic analysis of both pathogens showed genetic diversity among the Pakistani isolates as they clustered with isolates reported from reptiles, birds and ticks reported from worldwide countries. Risk factor analysis revealed that prevalence of Toxoplasma gondii varied between the screened lizard species. Highest parasite prevalence was observed in L. agrorensis (67%) and A. pannonicus (67%) followed by L. pakistanica (45%) and L. tuberculate (43%). All other studies risk factors varied non significantly with the prevalence of each parasite. In conclusion, this is the first study from Pakistan reporting a a very high Toxoplasma gondii while low Hepatozoon ophisauri prevalence in Pakistani lizards. We recommended that similar and large-scale studies must be conducted in various geo-climatic regions of Pakistan that are unexplored for the prevalence of these pathogens among the wild lizards as well as in other wildlife to broaden our knowledge about their genetic diversity, host-parasite interactions and effective control.