Inactivation of stationary phase Escherichia coli, Salmonella Typhimurium and Listeria innocua (10(8) CFU/ml) by high intensity pulsed electric fields (PEF) was studied in water and different buffers at pH 7.0. The fraction of survivors after PEF treatment with 300 pulses (5 Hz) of 26.7 kV/cm and a pulse width of 2 mus varied between 0.050% and 55%, but was always lower in Tris-HCl buffer than in HEPES-KOH buffer and water. When cell suspensions were stored for 24 h at 25 C after PEF treatment, the survivor fraction further decreased, except for E. coli in water and HEPES-KOH. By following the survival of untreated cells added to water or buffers that were previously PEF treated, this secondary inactivation could be ascribed to the formation of bactericidal components as a result of PEF treatment. Buffers and water containing 10 mM NaCl became bactericidal against all three bacteria upon PEF treatment, and the bactericidal effect could be neutralized by thiosulfate, suggesting that chlorine and/or hypochlorite had been formed. Also in the absence of Cl- ions, PEF treated water and buffers had bactericidal properties, but the specificity of the bactericidal effects against different bacteria differed depending on the buffer used. In the Ames mutagenicity test using His(-) S. Typhimurium mutant strains, PEF treated Tris buffers containing 10 mM Cl- ions, as well as PEF treated grape juice showed a mutagenic effect. The implications of these findings for the safety of PEF treated foods are discussed. (C) 2003 Elsevier B.V All rights reserved.