To manage pain effectively in people and animals, it is essential to recognise when pain is present and to assess its intensity. Currently there is very little information regarding the signs of post-surgical pain or its management in guinea pigs. Studies from other rodent species indicate that behaviour-based scoring systems can be used successfully to detect pain and evaluate analgesic efficacy. This preliminary study aimed to establish whether behaviour-based scoring systems could be developed to assess post-surgical pain in guinea pigs. This prospective, randomised, placebo-controlled study used 16 guinea pigs, and evaluated changes in behaviour following either anaesthesia alone or anaesthesia and orchiectomy. Behaviour was assessed using a combination of manual and automated scoring of remotely obtained video footage. A small number of behaviours were identified that appeared to have high specificity for pain caused by orchiectomy. However, the behaviours were displayed infrequently. The most common was a change in posture from standing to recumbency, sometimes with one hind leg extended either to the side or behind the body. A composite behaviour score incorporating these abnormal behaviours differentiated between the effects of surgery and anaesthesia alone (p<0.0001), and between animals that received analgesia post-operatively compared to an untreated group (p<0.0001). Although behavioural changes occurred in these guinea pigs after orchiectomy, the changes were relatively subtle and the individual specific pain-related behaviours occurred infrequently. However, it may prove possible to develop a behaviour-based scoring system for routine use in this species using a combination of pain-related behaviours.