Arboreal environments present considerable biomechanical challenges for animals moving and foraging among substrates varying in diameter, orientation, and compliance. Most studies of quadrupedal gait kinematics in primates and other arboreal mammals have focused on symmetrical walking gaits and the significance of diagonal sequence gaits. Considerably less research has examined asymmetrical gaits, despite their prevalence in small-bodied arboreal taxa. Here we examine whether and how free-ranging callitrichine primates adjust asymmetrical gait kinematics to changes in substrate diameter and orientation, as well as how variation in gait kinematics affects substrate displacement. We used high-speed video to film free-ranging Saguinus tripartitus and Cebuella pygmaea inhabiting the Tiputini Biodiversity Station, Ecuador. We found that Saguinus used bounding and half-bounding gaits on larger substrates versus gallops and symmetrical gaits on smaller substrates, and also shifted several kinematic parameters consistent with attenuating forces transferred from the animal to the substrate. Similarly, Cebuella shifted from high impact bounding gaits on larger substrates to using more half-bounding gaits on smaller substrates; however, kinematic adjustments to substrate diameter were not as profound as in Saguinus. Both species adjusted gait kinematics to changes in substrate orientation; however, gait kinematics did not significantly affect empirical measures of substrate displacement in either species. Due to their small body size, claw-like nails, and reduced grasping capabilities, callitrichines arguably represent extant biomechanical analogues for an early stage in primate evolution. As such, greater attention should be placed on understanding asymmetrical gait dynamics for insight into hypotheses concerning early primate locomotor evolution.