Our study purpose was to investigate the evolution of Babesia spp isolated from tissues of ticks that were found on 150 cows in Al-Diwaniyah province, Iraq. To fulfill the required purpose, sampling of 10 ticks was performed from each infested cow. These obtained ticks were morphologically recognized first, and then they were introduced to Lab investigation that was started with crushing the tick tissues to extract the genomic DNA of the Babesia spp. The DNA was then applied to polymerase chain reaction (PCR) method to recognize the amplification of the region that is related to the 18S rRNA gene. The resulted-amplified products were sequenced for the purpose of confirming and doing the phylogenetic analyses. Here, our study has demonstrated 2 different species according to the results of the sequencing and the phylogenetic analyses of the tested Babesisa species. These 2 species are SP1 and SP2. When the phylogenetic tree was built up, the results showed that SP1 and SP2 are closely related to Babesia bovis (HQ264126.1), an isolate from Texas, USA. Our study indicates interesting and valued data that could be used to study various aspects of the tick, Babesia species, and their control in Al-Diwaniyah City, Iraq.