The Rouge River is a 127-mile-long river located in the Metropolitan Detroit area of Southeastern Michigan. Due to its proximity to a highly urbanized location, pollution has frequently been a major issue concerning this river. This study aims to use quantitative polymerase chain reaction (qPCR) techniques to not only discover the most prevalent types of fecal-derived microbes in the Rouge River but to also look at the different antibiotic-resistant genes (ARGs), that is, antibiotic-resistant genes, found in these bacteria collected. At six sites along this river, field data and water samples were collected and brought back to the lab to undergo multiple tests, including DNA extraction, to profile fecal-derived bacteria found in the Rouge, and test to see if the bacteria that were found contained any ARGs. Overall, it was found that a plethora of bacteria in the Rouge River was from fecal matter, including strains such as Aeromonas, the most prevalent species found among the 6 sites, as well as certain strains of Clostridium. There was a lack of more notable, well-known, fecal-derived microbes, as strains such as Salmonella and Streptococcus were not found in any of the water samples. For ARGs, there were significant amounts of ARGs present within all the sites, with the Venoy site containing the highest percentage of ARGs, at 30%. Further analysis of the gene groups showed that the fluoroquinolone-resistant gene AAC (6)-Ib-cr was present throughout all six sites; further analysis shows that tetracycline ARGs are the most prevalent antibiotic ARGs in all six sites, coming in at 58%. Further research must be done, spanning the entire length of the river, to determine how widespread and detrimental these fecal-derived microbes and antibiotic-resistant genes are to the environment and to the people around them.