First Evidence of Haplotypes of Babesia bigemina From District Sialkot Pakistan and Their Relation to Other Countries

Abstract

Babesiosis is a tick borne disease of animals and causes significant losses to livestock sector and in turn to the economy of Pakistan. Objective: To analyze the molecular and phylogenetic analysis of Babesia species for inter and intra specific genetic variations in district Sialkot, Pakistan. Methods: Total 150 (Cow=120, Buffalo=20 and Sheep=10) blood samples were collected. Microscopic examination of blood smears stained by field stain method was performed. DNA was extracted by phenol chloroform extraction method and 18S rRNA gene was targeted for PCR amplification. MEGAX software was used to perform phylogenetic analysis. Haplotype analysis was performed with Popart software by using median joining network method. Results: Out of 150 blood samples, 50 (33%) were positive for Babesia bigemina (B. bigemina). Out of these, 36 showed 446bp bands specific for 18S rRNA gene of B. bigemina. In Phylogenetic tree one sequence from cow had 98% similarity with sequence from Uganda while all other sequences from cow had 98% similarity with sequence from China. Isolate from buffalo showed 98% similarity with sequence of buffalo previously reported from Pakistan and sequence of sheep appeared as more related to buffalo samples. Haplotype analysis from current study revealed four haplotypes, two from cow, one each from buffalo and sheep. Conclusions: This study was helpful for molecular and phylogenetic analysis of Babesia isolates from district Sialkot, Pakistan. Haplotyping has revealed the genetic variants among different and same host species.