Triclosan (TCS) is a broad-spectrum antimicrobial compound that is incorporated into numerous consumer products. TCS has been detected in aquatic ecosystems across the U.S., raising concern about its potential ecological effects. We conducted a field survey and an artificial stream experiment to assess effects of TCS on benthic bacterial communities. Field sampling indicated that TCS concentrations in stream sediments increased with degree of urbanization. There was significant correlation between sediment TCS concentration and the proportion of cultivable benthic bacteria that were resistant to TCS, demonstrating that the levels of TCS present in these streams was affecting the native communities. An artificial stream experiment confirmed that TCS exposure could trigger increases in TCS resistance within cultivable benthic bacteria, and pyrosequencing analysis indicated that TCS resulted in decreased benthic bacterial diversity and shifts in bacterial community composition. One notable change was a 6-fold increase in the relative abundance of cyanobacterial sequences and a dramatic die-off of algae within the artificial streams. Selection of cyanobacteria over algae could have significant implications for higher trophic levels within streams. Finally, there were no observed effects of TCS on bacterial abundance or respiration rates, suggesting that bacterial density and function were highly resilient to TCS exposure.