Bovine theileriosis, babesiosis and anaplasmosis are tick-borne hemoparasitic diseases and they are responsible for huge economic losses in livestock sector in Egypt. Currently, a total number of 110 dairy cows from different regions of Qena governorate, Egypt, were clinically and laboratory investigated for diagnosis of theileriosis, babesiosis and anaplasmosis using Giemsa stained blood film and Polymerase Chain Reaction (PCR) assay during the period from January 2019 to December 2019. On the basis of the obtained results, the overall prevalence of babesiosis, theileriosis and anaplasmosis among the screened cattle was 9.09%, 21.81%, and 25.45 respectively. Furthermore, mixed infections were seen in nineteen cases (17.27%) on the basis of blood film examination. PCR assay results revealed that, the infection rate with theileriosis, babesiosis and anaplasmosis was 5.45%, 7.27% and 16.36%, respectively. While, 7 cows were found to harbor a mixed infection. Additionally, hemato-biochemical alternations in theileriosis, babesiosis and anaplasmosis infected cows were also detected in this study. It could be concluded that PCR assay was the most sensitive test in the detection of the infection in all cases of the disease (acute, chronic and carriers) as once animals infected, they become carriers with low parasitemia after recovery and this low parasitemia cannot be detected by traditional examination of Giemsa stained thin blood smears.