Autotomy is a widespread phenomenon in which an organism will voluntarily lose a body part. In spiders, this behavior is typically used as a defensive strategy in which the spider will sacrifice a leg to escape a predator. However, losing a leg may expose the spider’s hemolymph to the environment, potentially providing an avenue for pathogens to infect the spider. The goal of this study was to determine if the loss of a leg has an immunological cost in the wolf spider, Tigrosa helluo. Spiders were collected on the St. Edward’s University campus at night. Each spider had their physical fitness determined by racing them down a plastic tube and recording their running speed. Spiders were assigned to either a control group that kept all of their legs, or a treatment group that had one randomly chosen back leg removed. Spiders in treatment groups were then further assigned to time intervals of ten minutes progressing up to a maximum of 60 minutes (ex. 10, 20,…60). Spiders designated in a treatment group with a leg removed had their wound packed with soil to introduce environmental pathogens, and were then acclimated for the appropriate time interval before hemolymph extraction; hemolymph was then extracted from all spiders. Protein assays indicated a significant difference among the time intervals with a spike in protein levels at 60 minutes post-autotomy. However, prophenoloxidase (PPO) showed no significant difference among time intervals, though PPO activity showed a trend of increasing at the 60-minute time point that requires further examination.