BackgroundStandardized and repeatable assessment of feline gait remains challenging, highlighting the need for controlled measurement systems such as pressure-sensitive treadmills.ObjectivesThis study aimed to compare kinetic and temporospatial gait parameters in healthy cats walking on a treadmill operated in standing vs. running mode. We hypothesized that no significant differences would be observed between modes.MethodsTen healthy adult cats were enrolled in a prospective, controlled crossover study. Inclusion criteria included body weight >4.0 kg, body condition score (BCS) 4–6/9, and age >24 months. Cats walked on a pressure-sensitive treadmill (FDM-T-CanidGait®, zebris Medical GmbH, Isny, Germany) under two conditions: standing mode (stationary belt) and running mode (moving belt). Five 2-min trials per mode were recorded. Kinetic and temporospatial gait parameters were assessed, including peak vertical force, vertical impulse, stride length, stance and swing phase duration, and step width. Right and left limb data were pooled and analyzed collectively as forelimbs (FL) and hindlimbs (HL). Data were analyzed using robust linear mixed-effects models (R 4.4.3), with significance set at p < 0.05.ResultsSignificant differences between treadmill modes were detected for peak vertical force and vertical impulse in both forelimbs and hindlimbs (p < 0.01). Forelimb swing and stance phase durations also differed significantly (p < 0.05). In contrast, stride length, step width, and hindlimb temporal parameters did not differ significantly between modes. Estimated differences were generally small in magnitude across variables.ConclusionIn healthy cats, stride length, step width, and hindlimb temporal parameters were comparable between standing and running treadmill modes. In contrast, kinetic variables (peak vertical force and vertical impulse) and forelimb stance and swing phase durations differed significantly between modes and should therefore be interpreted only within the same treadmill mode. These findings indicate that treadmill modes may be used interchangeably for selected temporospatial parameters but not for kinetic variables or forelimb temporal parameters.