Severin Blum ^1* Jana Gisler ¹ Emanuela Dalla Costa ² Stéphane Montavon ³ Claudia Spadavecchia ^1*

• ¹ Vetsuisse Faculty, University of Bern, Bern, Switzerland
• ² Department of Veterinary Medicine and Animal Sciences, Faculty of Veterinary Medicine, University of Milan, Milan, Lombardy, Italy
• ³ Veterinary Department of the Swiss Armed Forces, Bern, Switzerland, Bern, Switzerland

Study objective was to evaluate whether the application of a lip twitch could be proposed as conditioning stimulus in the context of a novel Conditioned Pain Modulation (CPM) assessment paradigm for use in horses.The study was a prospective, experimental, randomized trial. Twelve healthy horses were evaluated in two experimental sessions. The lip twitch was used as the conditioning stimulus in both sessions; electrical stimulation was used as the test stimulus in one session, while mechanical and thermal stimulations were used in the other. Differences between thresholds recorded before and during twitching (Δ) as well as their percent (%) change were computed for each stimulation modality as a measure of CPM. Heart rate and respiratory rate were recorded throughout the experiments to monitor physiological reactions, while the general level of stress and aversiveness toward twitching were scored using ad hoc behavioural scales. Based on these scores, interruption criteria were defined.Ten and seven horses completed the electrical and mechanical/thermal experimental sessions respectively. For electrical stimulation, median (IQR) Δ was -2.8 (-3.9, -1.1) mA and % change 87.9 (65.7-118.2) %; for mechanical stimulation, Δ was -18.2 (-6.4, -21.4) N and % change 343.5 (140, 365.3) %; for thermal stimulation, Δ was -3.1 (-9.2, -2.1) °C, while % change was not calculated. Heart rate and respiratory rates varied significantly over time, with higher values recorded during twitching. Median stress and aversion scores did not differ between the two sessions.