BACKGROUND: The UNESP-Botucatu multidimensional feline pain assessment scale (UFEPS) is a valid and reliable instrument for acute pain assessment in cats. However, its limitations are that responsiveness was not tested using a negative control group, it was validated only for ovariohysterectomy, and it can be time-consuming. We aimed to evaluate the construct and criterion validity, reliability, sensitivity, and specificity of the UFEPS and its novel short form (SF) in various clinical or painful surgical conditions. METHODS: Ten client-owned healthy controls (CG) and 40 client-owned cats requiring pain management for clinical or surgical care (20 clinical and 20 surgery group (12 orthopedic and eight soft tissue surgeries) were recruited. Three evaluators assessed pain, in real-time, in clinical cases before and 20 min after rescue analgesia and in surgical cases before and up to 6.5 hours postoperatively, by using the visual analog, numerical ratio, and a simple descriptive scale, in this order, followed by the UFEPS-SF, UFEPS and Glasgow multidimensional feline pain (Glasgow CMPS-Feline) in random order. For the surgical group, rescue analgesia (methadone 0.2 mg/kg IM or IV and/or dipyrone 12.5 mg/kg IV) was performed when the UFEPS-SF score was ≥4 or exceptionally according to clinical judgement. If a third interventional analgesia was required, methadone (0.1-0.2 mg/kg IM) and ketamine (1 mg/kg IM) were administered. For the clinical group, all cats received rescue analgesia (methadone 0.1-0.2 mg/kg IM or IV or nalbuphine 0.5 mg/kg IM or IV), according to the clinician in charge, regardless of pain scores. Construct (1-comparison of scores in cats undergoing pain vs pain-free control cats by unpaired Wilcoxon-test and 2-responsiveness to analgesia by paired Wilcoxon test) and concurrent criterion validity (Spearman correlation of the total score among scales), inter-rater reliability, specificity and sensitivity were calculated for each scale (α = 0.05). RESULTS: Reliability ranged between moderate and good for the UFEPS and UFEPS-SF (confidence intervals of intraclass coefficients = 0.73-0.86 and 0.63-0.82 respectively). The Spearman correlation between UFEPS and UFEPS-SF was 0.85, and their correlation with Glasgow CMPS-Feline was strong (0.79 and 0.78 respectively), confirming criterion validity. All scales showed construct validity or responsiveness (higher scores of cats with clinical and postoperative pain vs healthy controls, and the reduction in scores after rescue analgesia). The sensitivity and specificity of the UFEPS, UFEPS-SF and Glasgow CMPS-Feline were moderate (sensitivity 83.25, 78.60% and 74.28%; specificity 72.00, 84.67 and 70.00%, respectively). CONCLUSIONS: Both UFEPS and UFEPS-SF showed appropriate concurrent validity, responsiveness, reliability, sensitivity, and specificity for feline acute pain assessment in cats with various clinical and orthopedic and soft tissue surgical conditions.