Fatigue or stress fractures are an important clinical problem in humans as well as racehorses. An important question in this context is, when a bone experiences fatigue damage during extreme use, how much is it weakened compared to its original state? Since there are very limited data on this question and stress fractures are common in racehorses, we sought to determine the effect of fatigue loading on the monotonic strength of equine cortical bone. Beams were machined from the dorsal, medial and lateral cortices of the third metacarpal bones of six thoroughbred racehorses. Beams from left and right bones were assigned to control and fatigue groups, respectively (N = 18 each). The fatigue group was cyclically loaded in three-point bending at 2 Hz for 100,000 cycles at 0–5000 microstrain while submerged in saline at 37°C. These beams, as well as those in the control group, were then monotonically loaded to failure in three-point bending. The monotonic load-deflection curves were analyzed for differences using three-factor (fatigue loading, anatomic region, and horse) analysis of variance. The mean failure load was 3% less in the fatigue group, but this reduction was only marginally significant. Neither elastic modulus nor yield strength was significantly affected by the fatigue loading. The principal effects of fatigue loading were on post-yield behavior (yield being based on a 0.02% offset criterion). The work done and the load increase between yield and failure were both significantly reduced. All the variables except post-yield deflection were significantly affected by anatomic region. In summary, loading equivalent to a lifetime of racing does not significantly weaken equine cortical bone ex vivo. The clinical implication of this may be that the biological repair of fatigue damage can actually contribute to stress fracture if pressed too far.