This study examined the reliability of running performance across 3 nonmotorized treadmill (NMT) 5-km time trials (TTs) and physiological, gait, and perceptual differences at a 5-km pace for both NMT and motorized treadmills (MTs). Ten male runners experienced in road racing who had never run on an NMT completed 3 TT to establish personal best 5-km pace. In a later session, participants ran at this pace for 5 minutes on the NMT while metabolic, gait, and perceptual measures were recorded and then ran at outdoor 5-km personal best pace on an MT at 1% grade (counter-balanced crossover design). Intraclass correlation (ICC = 0.95) between the TT1 and TT2 was strong but improved between TT2 and TT3 (ICC = 0.99) with considerable reduction in variability. Nonmotorized treadmill resulted in a 24% slower pace (10.6 6 1.5 vs. 13.9 6 2.6 kmh21;p,0.001),shorterstridelength(1.0260.10vs.1.2760.18m;p,0.001),anddecreasedcadence(175612vs.181613stepsperminute; p = 0.01). However, V_ O2, respiratory exchange ratio (RER), lactate concentration, and heart rate did not differ between modalities (NMT = 3.4 6 0.4 Lmin21,0.9660.04,6.963.7mmol,172610bmin21; MT = 3.4 6 0.5 Lmin21,0.9660.04,5.763.4mmol,170610bmin21). rate of perceived exertion (RPE) for legs, breathing, and overall did not differ between treadmill types. A familiarization session should be included for TT using NMT. Other than gait and pace characteristics similar responses were elicited by both treadmills when running at 5-km pace. However, with these considerations, NMT TT of 4-km might be more appropriate in matching MT 5-km TT duration without altering physiological responses significantly