Energy cost of transport per unit distance (CoT; J·kg-1·km-1) displays a U-shaped fashion in walking and a linear fashion in running as a function of gait speed (v; km·h-1). There exists an intersection between U-shaped and linear CoT-v relationships, being termed energetically optimal transition speed (EOTS; km·h-1). Combined effects of gradient and moderate normobaric hypoxia (15.0% O2) were investigated when walking and running at the EOTS in fifteen young males. The CoT values were determined at eight walking speeds (2.4-7.3 km·h-1) and four running speeds (7.3-9.4 km·h-1) on level and gradient slopes (±5%) at normoxia and hypoxia. Since an alteration of tibialis anterior (TA) activity has been known as a trigger for gait transition, electromyogram was recorded from TA and its antagonists (gastrocnemius medialis (GM) and gastrocnemius lateralis (GL)) for about 30 steps during walking and running corresponding to the individual EOTS in each experimental condition. Mean power frequency (MPF; Hz) of each muscle was quantified to evaluate alterations of muscle fiber recruitment pattern. The EOTS was not significantly different between normoxia and hypoxia on any slopes (ranging from 7.412 to 7.679 km·h-1 at normoxia and 7.516 to 7.678 km·h-1 at hypoxia) due to upward shifts (enhanced metabolic rate) of both U-shaped and linear CoT-v relationships at hypoxia. GM, but not GL, activated more when switching from walking to running on level and gentle downhill slopes. Significant decreases in the muscular activity and/or MPF were observed only in the TA when switching the gait pattern. Taken together, the EOTS was not slowed by moderate hypoxia in the population of this study. Muscular activities of lower leg extremities and those muscle fiber recruitment patterns are dependent on the gradient when walking and running at the EOTS.