In Serbia, Babesia gibsoni in dogs is less common than Babesia canis. Although two clinical cases were reported a decade ago, no additional clinical reports have since been published. Recently, a co-infection of B. gibsoni and B. canis was documented in Austria following a dog’s trip to Serbia. The objectives of this study were to present comprehensive clinicopathological results of two clinical cases of B. gibsoni mono-infection in dogs in Serbia. Two male dogs: an 11-year-old Shih Tzu, and a 2-year-old Pit Bull Terrier, presented with clinical signs related to babesiosis with a history of biting by another dog. Both dogs had regenerative anemia, thrombocytopenia, and monocytosis while Shih Tzu had immune-mediated hemolytic anemia (IMHA). B. gibsoni mono-infection was confirmed by PCR testing. Both dogs were treated with a single dose of imidocarb-dipropionate, and a combination of metronidazole, clindamycin, and doxycycline (MCD protocol). The Shih Tzu also received prednisolone for three weeks. Following the MCD protocol, the Pitt Bull recovered, although thrombocytopenia persisted for nine months. In contrast, Shih Tzu's clinical condition worsened. The prednisolone treatment was discontinued, and azithromycin and atovaquone were introduced, leading to recovery after another three weeks of treatment. Long-term clinical and PCR monitoring revealed that the Pit Bull Terrier exhibited a more favorable response and a lower frequency of relapses compared to Shih Tzu. The findings suggest that B. gibsoni has become a clinically significant pathogen in Serbia. The MCD protocol appears effective for treating acute B. gibsoni infection in dogs, but further investigation is required to evaluate its efficacy in eliminating the parasite.