Theileriosis is a significant hemoprotozoal disease of domestic and wild ruminants in tropical and subtropical regions of the world. Ovines are mainly infected by T. ovis and T. lestoquardi, causing economic losses. Due to data scarceness in the Sulaymaniyah governorate, north of Iraq, this study was conducted to investigate subclinical theileriosis using microscopic examination and PCR. A total of 450 blood samples were collected from eight districts in Sulaymaniyah. The samples were randomly taken from clinically healthy sheep in 40 farms from April to Octo- ber 2017. Following the organism verification, PCR products were sequenced and aligned. The study results revealed that 76.0% (n=342) and 58.0% (n=261) of the examined samples were positive for Theileria spp. by PCR and microscopic examination. T. ovis was reported in 76.0% (n=342) of all tested samples, while T. lestoquardi was present in 28.4% (n=97) of the positive specimens. Higher infection rates were observed during July and August. Analysis of the 18S rRNA gene partial sequence of the studied isolates with corresponding sequences in GenBank showed high degrees of identities with T. ovis and T. lestoquardi isolates reported from Iraq and other countries. T. uilenbergi and T. sp. OT3 were detected only through analysis of obtained partial sequences from Theileria-positive samples. Following analysis, T. uilenbergi isolates represented a high homology degree with Theileria isolates from Iraq and China. The newly identified T. sp. OT3 showed >99% identity with T. sp. OT3 isolates of Chinese and Spanish origin.

Babesiosis is a parasitic disease caused by intraerythrocytic parasites of the genus Babesia, which infect both wild and domestic animals. Merozoite surface antigens (MSAs) have been identified as efficient immunogens in Babesia-infected animals. MSAs play a key role in the invasion process and have been proposed as potential targets for vaccine development. Epitope-based vaccines offer several advantages over whole protein vaccines as the immunogenic proteins are small and can induce both Th1 and Th2 immune responses, which are desirable for protection. However, the MSA, particularly gp45, is polymorphic in Babesia bigemina, posing a challenge to vaccine development. The purpose of this study was to develop a recombinant gpME (gp45-multi-epitope) for a vaccine against Babesia bigemina. B-cell, T-cell, and HLA epitope predictions were used to synthesize the gpME sequence from the consensus sequence of gp45. The gpME sequence was synthesized and cloned in the pET28α vector through the commercial biotechnology company to get pET28-gpME. The plasmid cloned with the gpME sequence comprising 1068 bp was expressed in a bacterial expression system. A band of 39 kDa of rec-gpME was obtained via SDS-PAGE and Western blotting. Rec-gpME @200ng was injected in calves 3 times at 2 weeks interval. The humoral response was evaluated through the indirect ELISA method. The ELISA with rec-gp45 protein showed a significant value of optical density. The recombinant protein containing multiple epitopes from the MSA gp45 may represent a promising candidate for a vaccine against Babesia bigemina.