Amblyomma hebraeum is the main vector of Rickettsia africae, the causative agent of African tick bite fever in southern Africa. Because pathogen dispersal is known to be influenced by tick adaptations to climate or host species, this study aimed to analyse the genetic diversity of A. hebraeum and R. africae infection of ticks collected from cattle in the Eastern Cape province of South Africa. DNA was extracted, amplified, and sequenced for the COI and ITS2 markers from A. hebraeum samples and the 17 kDa and ompA genes for rickettsial detection. Between six and ten haplotypes were identified from 40 COI and 31 ITS2 sequences; however, no population structuring was observed among sites (ΦST = 0.22, p < 0.05). All A. hebraeum isolates clustered with southern Africa GenBank isolates. Rickettsia africae was detected in 46.92% (95% CI = 41%–53%, n = 260) of ticks. All R. africae isolates clustered with strain PELE and Chucks, which were reported previously from South Africa. These results confirm that A. hebraeum populations are undergoing a recent population expansion driven by cattle movement, facilitating local and long dispersal events across the Eastern Cape province.
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