An increase in infections from non-fecal coliforms worldwide has caused concern that monitoring fecal coliforms overlooks pathogens that are not primarily associated with feces. A potential alternative to fecal coliforms is Staphylococcus aureus and methicillin resistant S. aureus (MRSA). The overall goal of the project was to quantify spatial patterns of an important opportunistic pathogen species of bacteria (S. aureus and MRSA) and to examine antibiotic susceptibility related to discharge into the Hillsborough River. There was a high presence of S. aureus and MRSA from all sites. Of the seven sites sampled, the site closest to Tampa General Hospital (site seven) had the lowest quantity of S. aureus but the highest percentage of MRSA. Whereas the site closest to The University of Tampa (site one) had the highest quantity of S. aureus and the lowest amount of MRSA. While Staphylococcus aureus was detected at significantly higher amounts during wet (rain) events, there was no correlation between these events and higher amounts of MRSA. These results suggest there is a potential correlation in the relationship of the quantity of S. aureus and antibiotic susceptibility (MRSA) of the isolates by location of discharge into the Hillsborough River. The results from this study are a good starting point to understanding the spatial patterns of S. aureus and MRSA in the Hillsborough River and gives us a platform for continued work.