BackgroundDistance walking fatigability (DWF) in people with multiple sclerosis (pwMS) is defined as a decrease in the distance walking over time. However, declines in gait quality (i.e., gait quality fatigability- GQF) may occur independently or alongside DWF. Objectivei) to investigate how walking fatigability manifests and its prevalence in pwMS; ii) to describe the temporal pattern of the changes of specific gait characteristics during the 6-minute walking test (6MWT) MethodsEighty-eight pwMS (EDSS 4[0–6.5], 49[21–70] years) and 47 healthy controls (HC- 46[25–60] years) performed the 6MWT wearing inertial measurement units. Gait characteristics (stride length, sensor-based gait speed, cadence, double support, step duration, stance phase, step duration asymmetry, step duration variability, foot-strike, toe-off, and leg circumduction) and walking distance were recorded in 1-minute intervals. A fatigability index was calculated by comparing the last and first minute of the 6MWT to identify abnormal worsening based on cutoff scores. The manifestation of walking fatigability was counted. The temporal pattern of worsening of gait characteristics during the 6MWT was examined in pwMS exceeding the cutoff values, compared to pwMS without abnormal changes and HC, using a two-way ANOVA (group vs. minutes) ResultsThirty-five pwMS presented both DWF and GQF, 2 presented isolated DWF, 27 presented isolated GQF, and 24 presented non-walking fatigability. PwMS having GQF presented worsening in gait characteristics (cadence, step duration, step duration variability, or toe-off angle) from minute 2 onwards of the 6MWT, while HCs and pwMS without abnormal changes stabilized gait from minute 2 towards the end of the 6MWT. ConclusionWalking fatigability in pwMS manifests not only as a decrease in walking distance but also as changes in gait quality. Understanding changes in gait characteristics during walking can help tailor rehabilitation interventions.