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Phytobenthos, major primary producers in freshwater, may be periodically exposed to herbicides through runoff, spray drift, and leaching, but experimental work on their response to herbicides is limited. Outdoor flumes were used to measure the recovery of intact phytobenthic biofilms, following a short-term pulse to a glyphosate-based herbicide (GBH) and chlorotoluron (CLT), singly or as a mixture (GBHC). Two days following the removal of the herbicides, significant declines in biomass and rates of areal gross maximum photosynthesis were recorded in GBH and GBHC-treated flumes. Herbicide pulses negatively impacted the biomass of phytobenthos, measured as chlorophyll a, which in turn may have alleviated resource limitation, evidenced by significant increases in biomass-specific rates of gross maximum photosynthesis. After 4.5 days, biomass concentrations were similar in controls and treatments, indicating biofilm recovery in the GBH and GBHC biofilms, though the proportion of green algae relative to diatoms was significantly higher in GBH and GBHC-impacted biofilms. Significant increases were recorded in the Trophic Diatom Index (TDI), indicating benthic diatom development along different trajectories, following the herbicide pulse. Collectively, these results indicate greater insights into herbicide impacts on phytobenthos may be gained by working with the whole phytobenthic community. [ABSTRACT FROM AUTHOR]