In South Asia, Theileria annulata is known to be less pathogenic to local breeds of Bos indicus cattle comparing to Bos taurus cattle and some of mix breeds between them. Seroepidemiological surveys have revealed high sero-prevalence of T. annulata in asymptomatic local breeds of cattle in Bangladesh. Therefore, these asymptomatic infection in local breeds can be infectious sources to more sensitive breeds. In this study, 59 bloods of cattle showing no symptom were screened by species specific PCRs for hemoto-protozoan parasites, to prove the existence of T. annulata parasite in asymptomatic cattle in Bangladesh. The T. annulata infection was confirmed along with other parasitic species, and this is the first report of T. annulata DNA detection in Bangladesh.