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Tick-borne pathogens (TBPs) are major constraints to livestock production and a threat to public health in Africa. This cross-sectional study investigated the risk of infection with TBPs in cattle of Lambwe Valley, Kenya. Blood samples of 680 zebu cattle from 95 herds in six geospatial clusters within 5 km of Ruma National Park were screened for bacterial and protozoan TBPs by high-resolution melting analysis and sequencing of PCR products. We detected <i>Anaplasma bovis</i> (17.4%), <i>Anaplasma platys</i> (16.9%)<i>, Anaplasma marginale</i> (0.6%), <i>Theileria velifera</i> (40%), and <i>Theileria mutans</i> (25.7%), as well as an <i>Anaplasma</i> sp. (11.6%) that matched recently reported <i>Anaplasma</i> sp. sequences from Ethiopia. <i>Babesia</i>, <i>Rickettsia,</i> and <i>Ehrlichia</i> spp. were not detected. The animal and herd-level prevalences for TBPs were 78.5% (95% confidence intervals (CI): 75.3, 81.5) and 95.8% (95% CI: 91.8, 99.8), respectively. About 31.6% of cattle were co-infected with 13 combinations of TBPs. The prevalence of TBPs differed between clusters and age, but the risk of infection was not associated with sex, herd size, or the distance of homesteads from Ruma. This study adds insight into the epidemiology of TBPs around Ruma and highlights the need for proactive surveillance of TBPs in livestock–wildlife interfaces.