We investigated the effects of transients in elution chemistry on bacterial desorption in water-saturated porous media. Two typical Gram-positive bacterial strains of Lactobacillus casei and Streptococcus mitis were used as the model bacteria in this research. These two strains were first deposited in the porous medium, after which the medium with deposited bacteria was flushed with rhamnolipid biosurfactant solutions with a step increase in concentrations, and pulse-type bacterial releases were obtained. Bacterial desorption was quantified from bacterial breakthrough curves. It was found that bacterial retention in silica sand corresponded to bacterial interaction free energies with silica sand evaluated at the equilibrium distance, which were calculated based on independently determined bacterial, sediment and solution surface thermodynamic properties. With the increase in rhamnolipid biosurfactant concentrations, interactions between bacteria and silica sand decreased, and consequently less bacteria were retained. The decrease in interactions between bacteria and silica sand with increasing rhamnolipid biosurfactant concentrations was attributed to a decrease in the solution electron acceptor parameter of the Lewis acid/base component of surface tension, gamma3+. The increase in rhamnolipid biosurfactant concentrations favored the decrease in solution gamma3+, and consequently decreased the interactions between bacteria and silica sand.