Despite its proven research applications, it remains unknown whether surface electromyography (sEMG) can be used clinically to discriminate non-lame from lame conditions in horses. This study compared the classification performance of sEMG absolute value (sEMGabs) and asymmetry (sEMGasym) parameters, alongside validated kinematic upper-body asymmetry parameters, for distinguishing non-lame from induced fore- (iFL) and hindlimb (iHL) lameness. Bilateral sEMG and 3D-kinematic data were collected from clinically non-lame horses ( n = 8) during in-hand trot. iFL and iHL (2-3/5 AAEP) were induced on separate days using a modified horseshoe, with baseline data initially collected each day. sEMG signals were DC-offset removed, high-pass filtered (40 Hz), and full-wave rectified. Normalized, average rectified value (ARV) was calculated for each muscle and stride (sEMGabs), with the difference between right and left-side ARV representing sEMGasym. Asymmetry parameters (MinDiff, MaxDiff, Hip Hike) were calculated from poll, withers, and pelvis vertical displacement. Receiver-operating-characteristic (ROC) and area under the curve (AUC) analysis determined the accuracy of each parameter for distinguishing baseline from iFL or iHL. Both sEMG parameters performed better for detecting iHL (0.97 ≥ AUC ≥ 0.48) compared to iFL (0.77 ≥ AUC ≥ 0.49). sEMGabs performed better (0.97 ≥ AUC ≥ 0.49) than sEMGasym (0.76 ≥ AUC ≥ 0.48) for detecting both iFL and iHL. Like previous studies, MinDiff Poll and Pelvis asymmetry parameters (MinDiff, MaxDiff, Hip Hike) demonstrated excellent discrimination for iFL and iHL, respectively (AUC > 0.95). Findings support future development of multivariate lameness-detection approaches that combine kinematics and sEMG. This may provide a more comprehensive approach to diagnosis, treatment, and monitoring of equine lameness, by measuring the underlying functional cause(s) at a neuromuscular level
