Bacterial resistance to antibiotics is a major obstacle that affects the health and economy of our society and continuous research in this area is crucial. The goal of our experiment was to assess antibiotic production in soil bacteria across three different location types: wooded, meadow, and sandy trail. Soil samples were collected from six locations in Central Park, NYC, and serial dilutions were plated and individual colonies counted. Colony forming units (CFU) were screened against six safe alternatives of ESKAPE pathogens. There was no difference in CFU among different substrate types, however, there was a significant difference in percent antibiotic producers across different substrate and culture media types. Future studies should consider investigating various soil substrates across temporal conditions including weather, season, and time of the day in order to determine optimal conditions for antibiotic producers.Our results demonstrate a point in time investigation and studies that examine temporal variation (across seasons, weather conditions, time of day, etc.) or expand on sample size and location (across multiple parks or cities) would further help identify ideal conditions for antibiotic producers and may help to identify novel producers. Unfortunately, we were not able to complete isolate identification (PCR and sequencing), and we plan to continue this work to determine the bacterial source of antibiotic production that we observed.