This study was aimed to determine the effect of age and gender on physiological variations in some hematological parameters and erythrocyte osmotic fragility in Kangal Shepherd dogs. A total of 32 clinically healthy Kangal Shepherd dogs were taken into the study and divided into 4 groups in equal numbers (Adult male = 8, Adult female = 8, Senior male = 8, Senior female = 8). Blood samples from the cephalic vein were taken into vacuum tubes containing EDTA by the technique and values of erythrocyte osmotic fragility and complete blood count were determined. The main effect of age and gender with age x gender interaction has no significant effect on hematological parameters (P>0.05). The main effect of age and age x gender interaction of erythrocyte osmotic fragility was statistically significant and higher in the senior females at 0.7% and 0.9% (in addition gender) NaCl concentrations compared to the other groups (P<0.05). However, there was no statistical difference between the adult female and adult male and senior male group at 0.7% and 0.9% NaCl concentrations (P>0.05). The findings may be useful for veterinary clinical practice in establishing Kangal shepherd breed-specific reference ranges for some important hematological parameters.