Genetic characterization of tick-borne pathogens in ticks infesting cattle and sheep from three South African provinces

Ticks are involved in the transmission of many public health and veterinary important pathogens. Although tick-borne pathogens are widely distributed in South Africa, information on tick-pathogen relationship needs to be updated particularly using modern molecular techniques. This study used PCR and sequencing to confirm the identity of the tick species collected from cattle and sheep from KwaZulu-Natal, Free State and Eastern Cape. Furthermore, presence of Babesia spp., Theileria spp., Anaplasma marginale, Rickettsia spp., Ehrlichia ruminantium and Coxiella burnetii was detected from tick DNA using species-specific PCR or nested PCRs. The study samples consisted of 390 adult ticks (male and female) which were pooled according to species, host animal and sampling site (three ticks per pool) for DNA extraction. The PCR results revealed that out of 130 tick DNA pools, 30 (23.1%) were positive for at least one pathogen. The most frequent pathogen was C. burnetii (9.2%), followed by Rickettsia spp. (7.7%), A. marginale (3.8%), T. mutans (3.1%), T. taurotragi (2.3%) and E. ruminantium (1.5%). The highest prevalence of pathogens was observed in ticks collected from cattle in Eastern Cape (16/42) and the lowest was in ticks obtained from sheep in Free State (1/21). Infected ticks were identified as Rhipicephalus evertsi evertsi (n = 13), R. appendiculatus (n = 3), R. decoloratus (n = 7) and Amblyomma hebraeum (n = 7). Coinfection with two pathogens was found in 21% of pathogen-positive pools. Analysis of Theileria taurotragi 18S rRNA, T. mutans 18S rRNA, C. burnetii htpB, Rickettsia spp. gltA, Rickettsia spp. ompA, E. ruminantium pCS20 and A. marginale Msp5 sequences showed that the pathogens detected in this study were genetically related to isolates previously reported in Africa. These findings provide important information on distribution of ticks and tick-borne pathogens of ruminants and will contribute in the formulation of future control strategies in South Africa.