Agricultural runoff can carry FIB that can pollute surface waters through the soil matrix. This study was designed to inspect the impact of temperature and matrix on the survival of FIB. The FIB were routinely enumerated over an 18-day period from fecal samples and freshwater microcosms maintained at 4oC, 22oC, and 35oC. It was found that the FIB studied underwent a primary growth of up to 1-log10 to 3-log10, highlighting the weakness of conventional FIB as indicators of pathogen contamination. The concentrations of FIB in the water phase were found to be significantly greater than those observed in the fecal phase in all FIB and their associated survivals were found to be significantly different too. Similarly, temperature was also found to be a significant factor for the survival of FIB. While the differences in the survival were significant, there was a slight variation in the patterns regarding the differences.