The low scientific attention in dealing with dog diseases is still present comparing to other animal species in Egypt in spite of its wide spreading as pets, hunting dogs or for other purposes. Canine babesiosis is a haemoprotozoal parasitic disease inducing significant clinical manifestations on the infected dogs. The present study aimed to accurately identify the causative species of canine babesiosis in naturally infected dogs in Egypt and its compatible tick vector. For this purpose 75 dogs including 19 dogs at Sharkia and 56 dogs at Dakahlia Governorates, Egypt were clinically examined and 20 (26.67%) dogs showed signs of fever, weakness, anorexia, pale mucous membrane and different degrees of anemia. During examination, there were 8 dogs had variant degrees of tick infestation. Further parasitological identification of 50 collected ticks showed that all the collected ticks were belong to Rhipicephalus sanguineus (R. sanguineus) species. Whole blood samples collected from all examined dogs were screened by thin blood film for presence of Babesia species piroplasm. Nine samples (12.0%) showed pyriform shaped protozoa inside red blood cells (RBCs). All blood samples and 5 pooled tick samples (10 ticks/each) were screened against 18s rRNA of Babesia species genome using semi-nested polymerase chain reaction (PCR). Babesia canis vogeli (B. canis vogeli) was the only detected species in fourteen blood samples (18.67%) and all collected tick samples. It is the first study in Egypt to use semi-nested PCR for detection of 18s rRNA of canine babesiosis in ticks collected from naturally diseased dogs.