First molecular evidence of reptile‐associated <i>Borrelia</i> spp. in tortoise bont ticks ( <i>Amblyomma marmoreum</i> ) collected in Zambia

Reptile-associated (REP) borreliae represent a distinct monophyletic lineage of spirochetes transmitted by hard ticks. These organisms have been reported in several regions worldwide, including South Africa, but had not been reported in ticks collected in Zambia until now. In this study, we screened for Borrelia spp. in Amblyomma marmoreum ticks collected from Leopard tortoises (Stigmochelys pardalis) in Southern Province, Zambia. Of the 87 tick DNA samples analysed, 74 (85.1%) were positive by Borrelia-specific real-time PCR. Subsequent BLASTn and phylogenetic analyses of flaB and 16S rDNA sequences confirmed that the detected Borrelia spp. belong to REP borreliae and are closely related to Borrelia sp. previously reported from South Africa. This study represents the first molecular confirmation of REP borreliae in A. marmoreum ticks collected in Zambia and expands the known distribution of these spirochetes. The findings underscore the need for further investigations on their ecology and epidemiology to better understand the public and animal health relevance and transmission dynamics of REP borreliae in reptile-associated tick populations.