Pain, a multifaceted condition associated with actual or potential tissue damage, transcends nociception and is characterised as a subjective, sensory, and emotional experience. Extensive literature describing the adverse effects of untreated post-surgical pain emphasises the necessity of a comprehensive pain management protocol, incorporating both pharmacological and non-pharmacological strategies to ensure successful patient outcomes. The present study aimed to determine whether a positive dog-owner interaction influences post-operative pain perception and stress (POPPS), as well as behavioural inactive rate variability in bitches that underwent elective surgery. Randomly selected bitches (n = 18) underwent ovariohysterectomy. Eight bitches experienced a 45-min visit post-surgery (VPS) characterised by positive dog-owner interaction, while the remaining ten did not (NVPS). Utilising the validated Short Form of the Glasgow Composite Measure Pain Scale (CMPS-SF) to assess acute pain in dogs via stress-related behaviours, a significant decrease in POPPS was evident in the VPS group after the 45-min dog-owner interaction at T3 (1 h after post-sedation recovery), in contrast to the NVPS group. CMPS-SF-associated descriptive items ‘Nervous/Anxious/Fearful’ and ‘Happy Content or Happy and Bouncy’ decreased and increased, respectively, with dog-owner positive interaction in the VPS group. The inactivity rate was significantly lower in VPS bitches after the post-surgery 45-min dog-owner interaction than in NVPS bitches. This preliminary study suggests that the owner’s presence reduces POPPS and may improve the dogs’ welfare while undergoing routine surgeries.