Several species of the tick-transmitted apicomplexans of the genus Theileria infect cattle and wildlife in Africa. Theileria (T.) parva is the cause of a disease that is a heavy burden for livestock production in Zimbabwe. ln recent years, the spatio-temporal patterns of theileriosis outbreaks in Zimbabwe have changed. lnvestigating the causative Theileria species and strains is key to understand the epidemiology and adapt contrai measures. Samples were collected from suspected bovine theileriosis cases (n=158) in two districts from November 2020 to May 2021. Frozen blood samples (n= 147) and organs (n=22) were tested by combining pan-Theileria 18S rRNA PCR, specific T. parva PCR and Sanger sequencing to determine the Theileria species present. Theileria was detected in 83.3% of the sampled clinical cases. T. parva (63%), T. velifera (24%), T. taurotragi (2.3%), T. mutans (2.3% ), and cc-infections (3.1 % ) were detected in the positive samples. To characterise the T. parva strains, we sequenced the sporozoite surface antigen p67 and the CD8+ target antigen Tp2 from 44 and 23 of the T. parva positive samples, respectively. Phylogenetic analyses showed that the T. parva strains are cattle-derived, clustering with the Kenyan Muguga isolate, which is part of the widely used T. parva vaccine cocktail, rather than with the Boleni strain, historically isolated and used for vaccine production in Zimbabwe. Dried blood spots (n=145) and faeces (n=87) were also screened by the 18S rRNA PCR to evaluate their suitability in Theileria diagnosis. Sensitivity was lower in dried blood spots (59.3%) and faeces (12.7%) compared to frozen blood. The use of non-invasive samples such as faeces can however be suitable to study the circulation of Theileria at population level, e.g. in a wild animal population. Overall, our data contribute to the knowledge of Theileria species circulating in Zimbabwe, informing contrai strategies and guiding future research.