The primary mechanical functions of limb bones are to resist deformation, and hence provide stiff levers against which muscles can act, and to be sufficiently strong to prevent breaking under static or dynamic loads which arise from normal and accidental activities. If bones perform these functions with a minimum amount of material, the energetic costs associated with building, maintaining and transporting the skeleton will be minimized. Appropriate skeletal architecture for minimizing mass while maximizing strength depends on forces imposed on structural elements. In the evolutionary acquisition of flight in the bat lineage, the forelimb skeleton must have come to experience locomotor-forces that differed from those engendered by the terrestrial locomotion of non-flying bat relatives. Here we successfully measure in vivo strain on the wing bones of flying mammals. Our data demonstrate that torsion and shear are unique and crucial features of skeletal biomechanics during flight, and suggest that the evolution of skeletal design in bats and other flying vertebrates may be driven by the need to resist these loads.