Fatty acid (FA) composition between biofilms and batch planktonic cultures were compared for two bacterial species Pseudomonas aeruginosa and Staphylococcus aureaus. Biofilm cultures exhibited decrease in saturated fatty acids (SAFA) that potentially conform to a more fluidic biophysical membrane property. The amount of FA in the biofilms' extracellular polymeric substance was not sufficient to consider it having a major contribution to the observed differences between biofilms and batch planktonic cultures. While biofilm grazing by the amphipod Hyalella azteca was evident, only certain bacteria-specific FA appeared to have the potential to be retained (odd-number SAFA and branched-chain FA). H. azteca with diet strictly consisted of bacteria biofilms did not demonstrate significant changes in their nutritional condition in terms of ω-3 and ω-6 polyunsaturated fatty acids (PUFA): combined with the results from fasting trials, H. azteca appears to have the capacity to retain ω-3 and ω-6 PUFAs up to 10 days.