Objective Canine babesiosis, an infectious disease transmitted by Dermacentor reticulatus, is exhibiting growing importance in Germany. The aim of this study was to display the increased incidence of canine babesiosis in the Rhine-Main area in Hesse, with special focus on the accumulation in the district of Gross-Gerau. Material and methods The retrospective study included dogs presented to the veterinary hospital between October 2018 and December 2020 and diagnosed with canine babesiosis on the basis of a positive Babesia spp.-PCR. Results A total of 697 dogs were tested by Babesia spp.-PCR during this time period. Of these, 81 (12 %) were positive. Sequencing was performed in 14 of the 81 dogs (17 %) (B. canis n = 13, B, vulpes n = 1). A simultaneous anaplasmosis infection was detected in 2 dogs. Strikingly, babesiosis cases occurred throughout the year with accumulations in March/April as well as in October. Evaluation of a complete blood cell count revealed pancytopenia in 44 of the 81 animals (54 %). Anemia was present in 66 (82 %), thrombocytopenia in 76 of the 81 patients (94 %). Only 2 of the 81 positive cases showed no hematological changes. Hyperbilirubinemia was found in 66 of 73 measured bilirubin levels (90 %). All animals were treated with two injections of imidocarb-diproprionate (Carbesia((R))) in 14-day intervals. Follow-up PCR was performed in 37 of the 81 patients (46 %). In the majority of cases (92 %), successful therapy was confirmed by a negative Babesia-PCR. A total of 6 of the 81 patients (7 %) were euthanized during the treatment period. The reasons for euthanasia were progressive renal disease, high-grade intravascular hemolysis necessitating multiple blood transfusions, and development of splenic and renal abscesses. Conclusion In dogs with clinical signs such as apathy, pyrexia and hemoglobinuria, as well as hematologic abnormalities comprising anemia, thrombocytopenia as well as pancytopenia, babesiosis needs to be included in the list of differential diagnoses. Testing should be initiated accordingly regardless of the season, however especially in spring and autumn.