Abstract Ten mixed-breed female dogs were used in the study. Abdominal wall lifting was performed with a sterilised cotton strip. Four portal sites were used to complete gasless laparoscopic ovariohysterectomy (GLOHE) procedures. The proper and suspensor ligament, ovarian pedicle, and broad ligament of the uterus, uterine artery and vein were coagulated using bipolar electrocoagulation. After the uterine body was coagulated and cut, the end of the cervix was ligated with a loop suture. Blood samples were obtained before the surgery, immediately after the procedure, and on 1, 3, and 5 d postoperatively for the determination of interleukin-6 (IL-6) and C-reactive protein (CRP). No significant surgical complications occurred. After gasless laparoscopic ovariohysterectomy, a significantly higher serum IL-6 level was found immediately after the surgery and 1 d postoperatively when compared with the level observed after traditional laparoscopic ovariohysterectomy (LOHE). However, no significant differences were observed in CRP level between GLOHE and LOHE. GLOHE proved to be a safe and feasible procedure in bitches.