The relationships between Escherichia (E) coli concentration, suspended particulate matter (SPM) particle size class, and land use practices are important in reducing the bacterium’s persistence and health risks. However, surprisingly few studies have been performed that quantify these relationships. Conceivably, such information would advance mitigation strategies for practices that address specific SPM size classes and, by proxy, E. coli concentration. To advance this needed area of research, stream water was sampled from varying dominant land use practices in West Run Watershed, a representative mixed-land use Appalachian watershed of West Virginia in the eastern USA. Water samples were filtered into three SPM intervals (<5 µm; 5 µm to 60 μm; and >60 μm) and the E. coli concentration (colony forming units, CFU) and SPM of each interval was quantified. Statistically significant relationships were identified between E. coli concentrations and size intervals (α < 0.0001), and SPM (α = 0.05). The results show a predominance (90% of total) of E. coli CFUs in the <5 μm SPM interval. The results show that land use practices impact the relationships between SPM and E. coli concentrations. Future work should include additional combined factors that influence bacterial CFUs and SPM, including hydrology, climate, geochemistry and nutrients.