The dataset includes the data of 64 selected specimens of ticks collected in Ruvuma Region, Tanzania, between November and December 2023, from which DNA was estracted. Data about: Tick morphological and molecular identification, molecular results for Piroplasmida, Anaplasmataceae, Rickettsia spp. and Borrelia were included. The data are part of a paper of the same authors entitled “Tick Diversity and Associated Pathogens in Livestock Systems of Southern Tanzania, and validation of a Pan-Borrelia TaqMan qPCR Assay “ submitted to the journal: Medical and Veterinary Entomology and under revision after referees evaluation.