Generation of bactericidal and mutagenic components by pulsed electric field treatment.

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 micros 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 degrees 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.

Activation and inactivation of Talaromyces macrosporus ascospores by high hydrostatic pressure.

The effect of high hydrostatic pressure treatment (with pressures of up to 700 MPa) on Talaromyces macrosporus ascospores was investigated. At 20 degrees C, pressures of > or = 200 MPa induced the activation and germination of dormant ascospores, as indicated by increased colony counts for ascospore suspensions after pressure treatment and the appearance of germination vesicles and tubes. Pressures of > 400 MPa additionally sensitized the ascospores to subsequent heat treatment. At pressures of > 500 MPa, activation occurred in a few minutes but was followed by inactivation with longer exposure. However, even with the most extreme pressure treatment, a fraction of the ascospore population appeared to resist both activation and inactivation, and the maximal achievable reduction of ascospores was on the order of 3.0 log10 units. Pressure-induced ascospore activation at 400 MPa was temperature dependent, with minimum activation at 30 to 50 degrees C and > or = 10-fold higher activation levels at 10 to 20 degrees C and at 60 degrees C, but it was not particularly pH dependent over a pH range of 3.0 to 6.0. Pressure inactivation at 600 MPa, in contrast, was pH dependent, with the inactivation level being 10-fold higher at pH 6.0 than at pH 3.0. Observation of pressure-treated and subsequently dried spores with the use of light and scanning electron microscopy revealed a collapse of the spore structure, indicating a loss of the spore wall barrier properties. Finally, pressure treatment sensitized T. macrosporus ascospores to cell wall lytic enzymes.