Understanding spatial patterns of Escherichia coli in freshwater sediments is necessary to characterize sediments as microbial reservoirs and to evaluate the impact of sediment resuspension on microbial water quality in watersheds. Sediment particle size distributions and streambed E. coli concentrations were measured along a 500‐m‐long reach of a first‐order creek 1 d before and on Days 1, 3, 6, and 10 after each of two artificial high‐flow events, with natural high‐flow events also occurring within the sampling periods. Spatial variability of E. coli was greater in sediments than in water within any given sampling; however, variation between sampling days was greater for water than for sediment. The mean relative difference analysis revealed temporally stable patterns of E. coli concentrations in sediments. Escherichia coli–rich locations along the reach corresponded to areas with higher organic matter and fine particle contents. Although low (k < 0.5 d−1) or negative survival rates were observed at most locations along the reach during times where no precipitation was recorded, a small number of locations showed such large concentration increase that on average the survival rate remained positive at the reach scale. The studied creek appears to have hot spots of concentration increase, where conditions for E. coli populations to increase are much more favorable than in most other locations across the reach. The effect of this increase can be seen at the reach scale but is difficult to discern without individual sampling that is dense in space and time. Core Ideas Temporally stable patterns of E. coli concentrations in sediments were observed. Reach‐average E. coli concentrations increased in sediments between high‐flow events. E. coli survival rates were generally low except in certain locations along the reach.

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