The presented study aimed to find out the differences between peripheral blood immune cell markers from healthy bitches and bitches with mammary tumors. Due to the fact that the role of canine innate immune cells in cancer remains poorly understood, the markers of innate cells were chosen for this research. Blood samples from female dogs with mammary tumors of epithelial and mesenchymal origin were investigated by flow cytometry. CD5 and CD11b markers of innate immune cells, phagocytic activity, and cellular killing were assessed. The number of CD11b lymphocytes was increased in tumors with epithelial origin. No significant differences were found between the percentages of phagocytic cells. However, the phagocytes of canine patients with tumors of epithelial origin showed increased phagocytosis compared to the control group. In oxidative burst test, a statistically significant difference between the number of reactive oxygen species (ROS) produced was demonstrated only between the group of bitches with epithelial tumors and the control group. These results may suggest that there are subpopulations of innate immune cells that may be involved in anti-tumor immune mechanisms and have a potential to be supportive diagnostic markers in canine mammary tumors.