Previous studies have shown that various short- and medium-chain free fatty acids (FFAs) and their corresponding monoacylglycerol esters (MGs) have antibacterial activity in vitro against primarily gram-positive bacteria. More recent studies have also shown that the growth of Helicobacter spp. is inhibited by linoleic acid and arachidonic acid. The purpose of the present study was to evaluate the susceptibility of Helicobacter pylori to the in vitro bactericidal properties of medium-chain MGs and FFAs. Incubation of H. pylori with saturated MGs, ranging in carbon chain length from C10:0 to C14:0, at 1 mM caused a 4-log-unit or greater reduction in the number of viable bacteria after exposure for 1 h. Lower levels of bactericidal activity were observed with C9:0, C15:0, and C16:0 MGs. In contrast, lauric acid (C12:0) was the only medium-chain saturated FFA with bactericidal activity against H. pylori. The MGs and FFAs were bactericidal after incubation for as little as 15 min at neutral or acidic pHs. Higher levels of MGs and FFAs were required for bactericidal activity in the presence of higher amounts of protein in liquid diets. We also found that the frequency of spontaneous development of resistance by H. pylori was higher for metronidazole and tetracycline (10(-5) to 10(-6)) than for C10:0 MG, C12:0 MG, and C12:0 FFA (< 10(-8)). Collectively, our data demonstrate that H. pylori is rapidly inactivated by medium-chain MGs and lauric acid and exhibits a relatively low frequency of spontaneous development of resistance to the bactericidal activity of MGs. Further studies are needed to establish whether MGs may be useful either alone or with other known therapeutic agents in the management of H. pylori infections in humans.