Hovering hummingbirds offer a model locomotor system for which analyses of both metabolism and flight mechanics are experimentally tractable. Because hummingbirds exhibit the highest mass-specific metabolic rates among vertebrates, maximum performance of hovering flight represents the upper limit of aerobic locomotion in vertebrates. This study evaluates the potential constraints of flight mechanics and oxygen availability on maximum flight performance. Hummingbird flight performance was manipulated non-invasively using air and gas mixtures which influenced metabolism via variable oxygen partial pressure and/or altered flight mechanics via variable air densities. Limits to the locomotor capacity of hovering ruby-throated hummingbirds (Archilochus colubris) were unequivocally indicated by aerodynamic failure in either air/helium or air/heliox mixtures. Air/helium mixtures are hypodense and hypoxic; failure to sustain hovering flight occurred at 63% of the density of sea-level air and at an oxygen concentration of 12%. Air/heliox mixtures are hypodense but normoxic; failure in hovering occurred at 47% of sea-level air density. Thus, hummingbirds demonstrated considerable power reserves in hovering flight as well as hypoxic tolerance. In air/helium mixtures, hovering was limited by oxygen supply and not by flight mechanics. Birds hovering in air/helium mixtures increased their mechanical power output but not their rate of oxygen consumption. By contrast, birds hovering in air/heliox mixtures increased both mechanical performance and metabolic expenditure. Under hypoxia, hovering hummingbirds demonstrated non-negligible, but still limited, capacities for anaerobic metabolism and/or oxygen storage. Depending on the physical context, hummingbird flight performance can therefore be limited by oxygen availability or by flight aerodynamics.