The purpose of this study was to determine if bleeding, for biomedical purposes, influenced the behavior of horseshoe crabs, Limulus polyphemus . In the summer of 2001, ten bled and ten control (unbled) female horseshoe crabs were tracked for 26 days using acoustic telemetry in a small estuary on Cape Cod, Massachusetts. All but three crabs, two bled and one control, were located during the study period. No mortality was observed in the control group, while 20% mortality was observed within the bled group. There was no significant difference in the average rate of movement or in the spatial distribution within the estuary between the two groups. However, a difference was detected in the movement patterns. Horseshoe crabs from the bled group had a random direction of movement compared to the directional movement pattern of the control group, suggesting that the bled crabs experienced more disorientation.