This study was conducted to evaluate the clinical relevance and effectiveness of ovariectomy (OVE) and ovariohysterectomy (OVH) in terms of postoperative pain behavior and surgical stress. Thirteen healthy intact mongrel purpose-bred female dogs were used in this study. OVE was performed in six dogs whereas OVH was performed in seven dogs. Prior to, 1, 2, 4, 6, 12, and 24 h after surgery, pain assessment was performed by using the Short form of Composite Measure Pain Scale (CMPS-SF) and blood analysis for the determination of glucose, creatine kinase (CK), and cortisol were done. Also, surgical time, duration of anesthesia, and incision length were compared between two groups. As a result, OVH group as opposed to OVE group showed significantly longer in surgical time, duration of anesthesia, and incision length. Also, based on the two-way ANOVA test, the CMPS-SF had significant differences (p < 0.05) between two groups, with the OVE dogs having lower values at 1, 2, 4, and 6 h postoperatively. In addition, in terms of CK, the value at 1, 2, 4, 6, and 12 h were significantly (p < 0.05) increased from the baseline value for the OVH group. However, as for CK in OVE group, the values at 4, 6, and 12 h were significantly increased from the baseline value. As for blood glucose, a significant (p < 0.05) increase from the baseline was shown at 1 h postoperatively in OVH group and no significant increase was shown in OVE group. In terms of serum cortisol level, the values at the 1 and 2 h were significantly (p < 0.05) increased from the baseline value for both groups. In conclusion, our study has shown that OVE can be considered as a superior choice in terms of sterilization technique in female dogs, considering shorter incision length, surgical time, duration of anesthesia along with lower pain and surgical stress response than OVH.