Sequencing-based phylogenetic-study of Babesia spp detected in tick tissues in Al-Diwaniyah province, Iraq

Abstract

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.

Molecular Diagnosis and Phylogenetic Analysis of Babesia Species Isolated From Ticks of Infested Cattle in Wasit Governorate, Iraq

“…We suggested that our strains might have descended from the ancestor of the Americanisolates, and that there are certain factors might play a role in distribution of this ancestor around the world such as moving of the animals and humans in addition to importation and exportation processes. Surprisingly, the local strains of our study were incompatible with that detected previously in cattle-feeding ticks in Al-Diwaniyah province, Iraq (11), demonstrating that there many strains of B. bovis circulate in Iraqi cattle livestock and their infested-ticks. Also, uncontrolled importation of domestic and wildlife animals might participate in existence of new isolates/strains in Iraq.…”

“…Findings of PCR assay indicated that both B. bovis and B. bigemina were circulating significantly in tick vectors. In Iraq, B. bovis was the only Babesia species isolated from tick samples of infested cattle (11). Therefore, this study was the first report of B. bigemina in ticks infested cattle.…”

“…In Iraq, tick-borne protozoa such as Babesia spp and Theileria spp represent an economic crisis for different animals (1, 7). For genus of Babesia or it species associated with ticks in cattle, there are only a few studies (11,26). This calls for further investigations on the distribution and diversity of Babesia species and their relevance to public and animal health in the region.…”

Molecular Characterization and Phylogenetic Analysis of Babesia Species Isolated from Domestic Cattle

“…But the constructed phylogenetic tree revealed that one sequence (B. bigemina Nar/Pak-8) clustered in a different sub clade, denoted to be genetically different from other isolates collected from Narowal. This tree was as similar as constructed within a study that conducted in South Africa in 2013 (Mtshali et al, 2013;Hajeel and Al-Fatlawi et al, 2019). The tree is drawn to scale, with branch lengths measured in the number of substitutions per site.…”

COX1 gene and ITS-2 region: A comparative study of molecular diagnosis of Parabronema skrjabini in camels (Camelus dromedaries), Al-Najaf Province, Iraq

“…Transferring the samples in a cold box to the parasitology lab of the College of Veterinary Medicine, Al-Qadisiyah University, gross examination of the samples grossly for collecting the adults, isolated worms were taken by forceps to diagnose species of parasites, depending on the diagnostic characteristics described by (1,15). Molecular identification of the nematodes uses extracted of 20 rDNA isolates and traditional PCR technique with higher purity of DNA (16)(17)(18), PCR master mix prepared by using AccuPower® PCR PreMix Kit and this master mix done according to company instructions included: DNA template 5µl, Forward primer 1.5µl, Reveres primer 1.5µl and PCR water 12µl. The PCR primers used in this analysis to the amplify ITS2rDNA gene of P. skrjabini, it is being designed according to (9), while COX1mtDNA gene was prepared as (non-degenerate primer) according to the molecular study of other Spirurida (19), these primers were produced from Bioneer company, Korea as following table 1, and reaction conditions of PCR for amplification showed as in the table 2.…”