State and federal agencies regulate fecal indicator bacteria (FIB), such as E. coli or Enterococcus, in order to manage public health risks at swimming beaches. Despite these goals, watershed managers are challenged in terms of how to best clean up sources of FIB because concentrations frequently exceed water quality objectives, and sources—both human and nonhuman sources of FIB—appear to be everywhere. Since most nonhuman fecal sources represent substantially lower public health risks than human sources do, this study utilizes the human fecal source marker HF183 to better define watershed managers’ riskiest sites and times in order to prioritize remediation actions. A total of 117 samples were collected and analyzed for both FIB and HF183 from 26 sites during multiple sampling campaigns between 2019 and 2021 along the mainstem in addition to major tributaries in a highly urbanized watershed. The results indicated that the vast majority of samples (96%) quantified HF183 during wet weather, ranging from 99 to 44,768 gene copies/100 mL. Similar to HF183, the FIB results exceeded water quality objectives for 100% of the samples in wet weather; however, HF183 was rarely quantified in dry weather, with 3 of 72 samples (4%) exceeding 500 gene copies/100 mL, while two-thirds of samples (67%) exceeded FIB water quality objectives during dry weather. Where HF183 was detected in dry weather, isolated and unpredictable events explained human fecal pollution. It is more challenging in wet weather to identify and quantify the source(s) of human fecal pollution.