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The relationship between membrane damage and loss of viability following pressure treatment was examined in Escherichia coli strains C9490, H1071, and NCTC 8003. These strains showed high, medium, and low resistance to pressure, respectively, in stationary phase but similar resistance to pressure in exponential phase. Loss of membrane integrity was measured as loss of osmotic responsiveness or as increased uptake of the fluorescent dye propidium iodide. In exponential-phase cells, loss of viability was correlated with a permanent loss of membrane integrity in all strains, whereas in stationary-phase cells, a more complicated picture emerged in which cell membranes became leaky during pressure treatment but resealed to a greater or lesser extent following decompression. Strain H1071 displayed a very unusual pressure response in stationary phase in which survival decreased to a minimum at 300 MPa but then increased at 400 to 500 MPa before decreasing again. Membranes were unable to reseal after treatment at 300 MPa but could do so after treatment at higher pressures. Membrane damage in this strain was thus typical of exponential-phase cells under low-pressure conditions but of stationary-phase cells under higher-pressure conditions. Heat shock treatment of strain H1071 cells increased pressure resistance under low-pressure conditions and also allowed membrane damage to reseal. Growth in the presence of IPTG (isopropyl-␤-D-thiogalactopyranoside) increased resistance under highpressure conditions. The mechanisms of inactivation may thus differ at high and low pressures. These studies support the view that membrane damage is an important event in the inactivation of bacteria by high pressure, but the nature of membrane damage and its relation to cell death may differ between species and phases of growth.

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The influence of several factors (amplitude of ultrasonic waves, external static pressure, temperature and viscosity of medium) acting, either individually or in combination, on the amount of power transferred to a liquid medium during ultrasonication (power output) was measured by calorimetry. At constant amplitude (150 microns) and pressure (200 kPa), the power output decreased as the temperature was raised. The effect of temperature could be compensated by increasing pressure. The magnitude of the increase in power output due to raising the pressure depended on the pressure range and the treatment temperature. At all temperatures and pressures studied, the power output increased exponentially when the amplitude was increased linearly. The magnitude of this power output did not depend on the temperature or pressure of treatment. At 40 degrees C the magnitude of the increase in power output due to increasing the pressure was not influenced by the amplitude of sonic waves. The power output increased as the viscosity of the medium was increased. The magnitude of this effect did not depend on the amplitude but on the static pressure.