This study was done to evaluate changes in bacterial communities from a stream in the Cascade Cave system, Carter Caves State Resort Park in Kentucky (United States). Samples from cave sediments, cave water, and from Bio-Traps(R) were acquired from July 2013 through December 2013, from two locations separated by approx. 1.5 km, or approx. 12 hours water flow travel time. Bacterial communities were assessed from amplicon pyrosequence libraries of single-direction reads generated using multiplexed, bar-coded primers targeting the V1-V3 region of the bacterial of 16S rRNA gene.