The ventral midline approach (VMA) and right flank approach (RFA) are common procedures for the sterilization of bitches. This study compared the different parameters viz. total duration of surgery, recovery time, and length of the incision as well as body temperature, heart rate, respiration rate, and SpO2 in each approach. Twenty (20) bitches were divided randomly for the RFA and VMA. Meloxicam (0.2 mg/kg) was administered subcutaneously half an hour before the induction to provide preemptive analgesia. Diazepam and ketamine were administered intravenously at dose rates of 0.25 mg/kg and 2.5 mg/kg, and 0.17 mg/kg and 3.33 mg/kg, respectively to produce and maintain anesthesia. Each parameter was recorded at the pre-operative, operative and post-operative times. The average duration of surgery and length of incision of RFA (16.1 ± 5.13 min and 2.44 ± 0.83 cm) were significantly lower (p < 0.05) than the VMA (21.3 ± 5.48 min and 3.53 ± 0.7 cm). The operated bitches showed hypothermia (p < 0.05) at 1 hour compared to baseline and 24 hours of surgery. Heart and respiration rates increased significantly (p < 0.05) during traction and severing of ovarian ligaments in bitches within the RFA group, but there was no significant difference within VMA approaches. The sedation score was significantly higher (p < 0.05) at 1 hour after surgery in both approaches. Based on the duration of surgery and length of incision RFA approach was quick and minimal skin wound. Further studies on bitches considering molecular investigations of surgical stress are imperative.