In the present study hematological and biochemical parameters of males and females of three indigenous chicken breeds were investigated. For this total 120 mature bird, forty birds (20 male and 20 female) from each indigenous breeds Aseel, Kadaknath and Rajasri were used. A significantly (p<0.05) higher RBC, haemoglobin and PCV values were recorded in males than females in Aseel and Kadaknath breeds. The difference in WBC count (103/µl) between Kadaknath male and female was significant (p<0.05) but was insignificant in Aseel and Rajasri, however there was a significant (P<0.05) difference in their lymphocyte and heterophil counts in Aseel. Within erythrocyte indices, significantly (P<0.05) higher values for MCV and MCH was noticed in Rajasri males than females. There was significantly (p<0.05) higher MCHC recorded in Kadaknath males than females. A significantly (P<0.05) higher total protein values observed in females than males of all the three breeds. The serum cholesterol values differed significantly (P<0.05) among the males and females of Aseel but not among the other two breeds. A significantly (P<0.05) high serum bilirubin (mg/dl) levels were noticed in Aseel females than males. A significantly (P<0.05) higher SGOT levels were observed in female Kadaknath and male Rajasri birds. A significantly (P<0.05) higher SGPT levels were observed in males than females of Rajasri. There were significantly (P<0.05) higher serum calcium levels observed in males than females of all the three breeds. There was a significant (P<0.05) difference observed in serum phosphorus(mg/dl) levels among the male and female of Rajasri. Kadaknath female had significantly (p<0.05) higher blood glucose level than male.