This study was designed to conduct phylogenetic analysis on the cytochrome b and TaPIN1 genes of Theileria annulata and to detect potential associations between mutations in these two genes and resistance to buparvaquone. Positive PCR products for the cytochrome and TaPIN1 genes of two samples were sequenced and phylogenetically analysed. When cytochrome b gene sequences were aligned to the reference isolate (XM949625), the phylogenetic analysis showed the presence of two new T. annulata cyt b gene isolates. These isolates showed 99.9% homology with each other and exhibited high homology with existing Egyptian and global isolates and were deposited in GenBank under accession numbers (PP920503) and (PP920504) for samples from Alexandria and Beheira governorates, respectively. Also, there is one non-synonymous mutation in our two isolates in codon 146 (Alanine146Threonine), which is in the Qo1 region (one of the buparvaquone binding sites). When TaPIN1 gene sequences were aligned to the reference isolate (XM949502), the phylogenetic analysis showed the presence of two new T. annulata TaPIN1 gene isolates. These isolates showed 98.1% homology with each other and showed high homology with global isolates. These two isolates are the first Egyptian Theileria annulata TaPIN1 gene isolates and were deposited in GenBank under accession numbers (PP920505) and (PP920506) for samples from Alexandria and Beheira governorates, respectively. Moreover, phylogenetically, Also, there are two non-synonymous mutations in these two isolates in codon 24 (Isoleucine24Valine) and 52 (Methionine52Threonine), which are related to resistance against buparvaquone. Briefly, this is the first Egyptian study that investigated the TaPIN1 gene of Theileria annulata and the first study in Egypt that investigated both the TaPIN1 and cytochrome b genes in the same samples. This study has introduced evidence of point mutations in the cytochrome b and TaPIN1 genes of T. annulata that might be associated with buparvaquone treatment failure in Egypt.
Key words: Theileria annulata, Cytochrome b gene, TaPIN1 gene, Phylogenetic analysis
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