Phylogenetic Analysis and Detection of Drug Resistance Gene in Theileria annulata Isolated from Buffaloes

Abstract

Bovine theileriosis, caused by Theileria annulate, is disease affecting cattle and buffaloes worldwide. The current study aimed to screen the blood samples of 30 naturally suspected local buffaloes infected with Theileria species. The blood samples were initially examined by light microscopic and then the positive samples were subjected to PCR reactions. All 30 animals indicated clinical symptoms, such as high fever, loss of appetite, the presence of the hard tick, and enlargement of lymph nodes. The amplified products of 18S rRNA were analyzed, along with molecular detection of the drug-binding site alterations and interrelated changes in the cytochrome b (cyto b) gene. Blood smears revealed the presence of infected erythrocytes with Theileria spp. The PCR results confirmed infection in samples when DNA amplified with partial 18S rRNA and cyto b genes. The sequencing data were obtained from GeneBank using the accession numbers OM937770.1, ON207523.1, ON207525.1, ON207524.1, ON207526.1, and ON207527.1 Following BLAST analysis (Basic Local Alignment Search Tool), genetic differences were observed between the Iraqi isolate OM937770.1 and strains from India, Iran, and Turkey. The data obtained from the current study may reveal the genetic alteration of the local strain in the drug-target codons, which are found in one isolate and are different from the GenBank isolates. The results suggest that the failure of buparvaquone therapy might be due to the resistance to cyto b gene.