Surface-adherent microcolonies of Listeria monocytogenes were obtained by growing cells on glass slides immersed in a low nutrient medium containing excess glucose. The susceptibility of the adherent populations to benzalkonium chloride (100, 400, and 800 ppm solutions), anionic acid sanitizer (200 and 400 ppm solutions), and heat (55 and 70°C) was determined. Adherent microcolony cells decreased by 2 to 3 log cycles immediately after exposure to the sanitizers. The remaining population of microcolony cells survived 20 min of exposure demonstrating resistance to both sanitizers at all concentrations. Adherent single cells exhibited an initial 3 to 5 log decline in numbers and reached undetectable levels after 12 to 16 min of exposure to the sanitizers. Planktonic cells were reduced to undetectable levels after 30 sec exposure to the lowest concentration of each sanitizer. Removing adherent cells from the surface increased their sanitizer susceptibility to near that of planktonic cells. Heating adherent microcolonies at 70°C for 5 min resulted in a less than 5-log decrease in population with a surviving population of over 10 cfu/sq cm. These results demonstrate the ability of L. monocytogenes to develop resistance to inactivating agents when exposed to specific growth environments.