The ability of different molar-mass fractions of interstitial dissolved organic carbon (DOC) from a small stream to stimulate bacterial growth was determined. Growth was the greatest in the low-molar mass (<1 kDa) (LMM) fraction and the least in the high-molar mass (>10 kDa) (HMM) one. The LMM fraction consistently had a higher ratio of carbon to organic nitrogen than other fractions. The fluorescence in situ hybridization analysis revealed that bacterial populations in interstitial water were dominated by beta-Proteobacteria. gamma-Proteobacteria together with the Cytophaga-Flavobacteria cluster dominated in LMM DOC (66.3%) while the proportions of bacterial groups in HMM and medium-molar mass (MMM) fractions were more equal. The fact that gamma-Proteobacteria strongly dominates in LMM DOC fraction refutes the idea that LMM DOC is readily utilized by all heterotrophic bacteria.