PCR-Based Detection of Babesia ovis in Rhipicephalus bursa and Small Ruminants

Abstract

This study aimed to assess the prevalence of Babesia ovis infection in adult Rhipicephalus bursa and small ruminants in West Azerbaijan province, Iran. Blood samples were collected from 280 sheep and 122 goats of forty randomly selected flocks. Specific B. ovis fragment was detected in 67 animals (16.7%), of which 52 animals (18.6%) were sheep and 15 animals (12.2%) goats (P < 0.05). Of the 848 R. bursa collected from naturally infested small ruminants and farm dogs, Babesia ovis was detected by PCR in salivary glands of 94 adult ticks. The frequency of B. ovis infection was higher in flocks with tick in comparison with animals without tick (P < 0.05). Positive amplification from blood of ruminants, ticks, oviposition ticks, eggs, and larvae was subjected to restriction digestion with HphI. One RFLP profile was produced. The PCR-RFLP results indicated that one strain of B. ovis exists in this area. The results showed that the PCR was useful method to investigate the epidemiology of small ruminants' babesiosis. Furthermore, R. Bursa, which can transovarially transmit B. ovis and as well as being widely distributed in West Azerbaijan province, Iran, might play an important role in the field as a natural vector of B. ovis.

Figure 1

PCR products amplified using B. ovis-specific primers. Lane M: 50 bp DNA ladder (Fermentas, Germany), lane 1: infected sheep blood, lane 2: infected goats blood, lane 3: negative control, lane 4: positive control, lane 5: tick collected from sheep, lane 6: tick collected from goat, lane 7: tick collected from dogs, lane 8: tick sample after oviposition, lane 9: egg sample, and lane 10: larvae sample.

Figure 2

PCR and RFLP profile of amplified 549 bp fragment of the B. ovis-specific ssu rRNA gene. Lane M: 50 bp DNA ladder (Fermentas, Germany), lane 1: undigested PCR product, lane 2: digested PCR product from infected sheep, lane 3: digested PCR products from infected goats, lane 4: tick collected from sheep, lane 5: tick collected from goat, lane 6: tick collected from dogs, lane 7: tick sample after oviposition, lane 8: egg sample, and lane 9: larvae sample.