Molecular characterization of Theileria species infecting domestic ruminants

Abstract

Present study was carried out with an aim to record the prevalence and characterization of Theileria species infecting domestic ruminants in Andhra Pradesh. newlinePCR was standardized for detection of Theileria genus specific, T. annulata, T. orientalis, T. lestoquardi and T. ovis by targeting SSUr RNA (1098 bp), 30 kDa Major T.annulata merozoite surface protien (721 bp), Major piroplasms surface protein (776 bp), 30 kDa Theileria lestoquardi merozoite surface antigen (785 bp) and SSU rRNA (520 bp), respectively. The concentration and average range of genomic DNA yield obtained from cattle, buffaloes, sheep and goats was 85.79 - 98.76, 87.26 - 95.42, 86.72 - 94.76 and 94.86 to 101.72 ng/µl, respectively. The and#955;260/and#955;280 range was recorded as 1.6 and above for extracted DNA samples. Out of 1426 blood samples, Theileria DNA was found in 185 (12.97 %) samples. Among 534 cattle tested, 102 (19.10%) were found to be positive for Theileria spp. The results for buffalo blood samples were only 18 (5.42%) out of 332 tested. Theileria spp. positivity was seen in 61(15.01%) and 4 (2.56%) out of 404 and 156 sheep and goats screened, respectively. All the 185 Theileria positive isolates by genus specific PCR were subjected to Theileria species specific PCR. Out of 120 Theileria positives from cattle and buffaloes, 97 isolates were confirmed as T. annulata and the remaining 23 of T. orientalis. The amplified PCR product size of 721 bp confirmed the presence of T. annulata in 82 and 15 cattle and buffalo isolates, respectively. A product size of 776 bp confirms the presence of T. orientalis DNA in 20 and 3 cattle and buffalo isolates, respectively. A total of 65 blood samples of sheep (n=61) and goats (n=4) were detected positive for Theileria spp. by PCR. Prevalence of T. ovis was confirmed in 35 and 2 isolates of sheep and goats, respectively with PCR by targeting SSU rRNA gene primers and amplified product size of 520 bp. Similarly, prevalence of T. lestoquardi in 26 and 2 isolates of sheep and goats, respectively was confirmed with