Determination of Rhipicephalus spp. as vectors for Babesia ovis in Iran

The tick-borne diseases of livestock constitute a complex of several diseases with different etiological agents, such as protozoa, rickettsia, bacteria, and viruses. One problem discussed in protozoan infection is the determination and characterization of the transmitter agent. Because many analyses were performed with the salivary gland smears using the methyl-green-pyronin staining method or the Feulgen staining method, the transfer vector remains unanswered in some cases. The aim of this study is to recognize Babesia ovis in the salivary gland of Rhipicephalus spp. using polymerase chain reaction (PCR). Deoxyribonucleic acid (DNA) was isolated from 269 salivary gland of Rhipicephalus spp. (108 R. bursa, 87 R. turanicus, 74 R. sanguineus) collected from sheep with suspected to babesiosis. The isolated DNA was then analyzed with the primers derived from the hypervariable region V4 of 18S ribosomal ribonucleic acid (rRNA) of the Babesia species. For the specificity of the PCR product and discriminating from Babesia motasi and Babesia crassa, nested PCR and restriction fragment length polymorphism was performed. As positive control for the DNA extraction procedure, the DNA was analyzed with the common primers designed from the 18S rRNA of the Ticks (Rhipicephalus, Hyalomma, Haemophysalis, Dermacentor, Ixodes, Boophilus). B. ovis was detected in salivary gland of 18.5% R. bursa, 9.1% R turanicus, and 8.1% R. sanguineus, respectively.