Since the 1980’s production of the number of new antibiotics has been in rapid decline. In our research we address this issue by screening antibiotic producers and optimizing the conditions for antibiotic production. CR3-9 (Rheinheimera chironomi), a bacterium isolated from the Kinnikinick River in River Falls, WI, was used in this study. The focus of this research is to test the effect of physical stressors on CR3-9 and increase its antibiotic activity. Methods included testing media types, using agitation (100 rpm, 200 rpm) and different incubation temperatures (25℃, 28℃, 30℃). Results indicated that CR3-9 produced increased antibiotic activity against Pseudomonas putida when cultured in Potato Dextrose Broth and was incubated at 25℃ with an agitation of 200 rpm for 48 hours. Future research efforts will focus on the effects of chemical conditions, such as changing the pH (5.0, 6.0, 7.0) of culture media, and altering carbon and protein sources.