P33 surface protein gene of Theileria sergenti was amplified by PCR,and ligated into pMD18-T vector.The recombinant plasmid was certified by PCR amplification,digested with endonuclease and sequenced.A prokaryotic expression plasmid was constructed by inserting the P33 surface protein gene into pGEX-4T-3.The expression was induced by IPTG,and the expressed protein was analyzed by SDS-PAGE and Western blotting.In results,the BC-48 cDNA was an ORF which was 868 bp in length and encoding 292 amino acids.Similary of nucleotide sequence between the cloned P33 surface protein gene and China strain was 99.1%.The fusion protein of 59 000 in size was recognized by positive serum of cow Theileria sergenti,indicating that the fusion protein had a strong immunity.
Ovine theileriosis is a tick-borne disease that restricts the development of small ruminant husbandry in northern China. In this study, we report on a molecular epidemiological survey of ovine Theileria spp. in 198 blood samples taken from sheep in northern China. The DNA samples were screened by a nested polymerase chain reaction (PCR) targeting the 18S rRNA gene of ovine Theileria spp. The prevalence of ovine Theileria spp. in Yanji, Nongan, Longjing, Toudao and Jinchang was 80%, 40%, 37%, 24% and 32%, respectively. The sequencing analyses approved the present of the T. orientalis and/or T. luwenshuni in these regions. Taken together, we have demonstrated a high incidence of Theileria spp. in northern China that calls for the need to design effective control programs for ovine theileriosis.
In the present study, a total of 137 blood samples were collected from cattle and water buffaloes in central region of Vietnam and tested using nested polymerase chain reaction (nPCR), enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT) to determine the molecular and serological prevalence of Babesia bovis and Babesia bigemina. In cattle, the prevalence of B. bovis and B. bigemina was 21.3% and 16.0% by nPCR, 73.4% and 42.6% by ELISA and 60.6% and 59.6% by IFAT, respectively, whereas those of water buffalos were 23.3% and 0% by nPCR, 37.2% and 9.3% by ELISA and 27.9% and 18.6% by IFAT, respectively. IFAT and ELISA detected a higher number of infected cattle and water buffaloes than nPCR totally. Statistically significant differences in the prevalence of the two infections were observed on the basis of age. Overall, the current data suggest high incidence of B. bovis and B. bigemina infections in the central region of Vietnam, which is needed to develop comprehensive approach to the modern surveillance, diagnosis and control of bovine babesiosis.