P. Mañas

Influence of different factors on the output power transferred into medium by ultrasound

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.

Membrane damage and microbial inactivation by chlorine in the absence and presence of a chlorine-demanding substrate.

ABSTRACT The relationship between cell inactivation and membrane damage was studied in two gram-positive organisms, Listeria monocytogenes and Bacillus subtilis, and two gram-negative organisms, Yersinia enterocolitica and Escherichia coli, exposed to chlorine in the absence and presence of 150 ppm of organic matter (Trypticase soy broth). L. monocytogenes and B. subtilis were more resistant to chlorine in distilled water. The addition of small amounts of organic matter to the chlorination medium drastically increased the resistance of both types of microorganisms, but this effect was more marked in Y. enterocolitica and E. coli. In addition, the survival curves for these microorganisms in the presence of organic matter had a prolonged shoulder. Sublethal injury was not detected under most experimental conditions, and only gram-positive cells treated in distilled water showed a relevant degree of injury. The exposure of bacterial cells to chlorine in distilled water caused extensive permeabilization of the cytoplasmic membrane, but the concentrations required were much higher than those needed to inactivate cells. Therefore, there was no relationship between the occurrence of membrane permeabilization and cell death. The addition of organic matter to the treatment medium stabilized the cytoplasmic membrane against permeabilization in both the gram-positive and gram-negative bacteria investigated. Exposure of E. coli cells to the outer membrane-permeabilizing agent EDTA increased their sensitivity to chlorine and caused the shoulders in the survival curves to disappear. Based on these observations, we propose that bacterial envelopes could play a role in cell inactivation by modulating the access of chlorine to the key targets within the cell.

Occurrence of sublethal injury after pulsed electric fields depending on the micro-organism, the treatment medium ph and the intensity of the treatment investigated

Aims:  The objective was to investigate the occurrence of sublethal injury after pulsed electric field (PEF) depending on the treatment time, the electric field strength and the pH of the treatment media in two Gram‐positive (Bacillus subtilis ssp. niger, Listeria monocytogenes) and six Gram‐negative (Escherichia coli, Escherichia coli O157:H7, Pseudomonas aeruginosa, Salmonella serotype Senftenberg 775W, Salmonella serotype Typhimurium, Yersinia enterocolitica) bacterial strains.

Inactivation of Salmonella Enteritidis, Salmonella Typhimurium, and Salmonella Senftenberg by ultrasonic waves under pressure.

The resistance of Salmonella Enteritidis (ATCC 13076), Salmonella Typhimurium (ATCC 13311), and Salmonella Senftenberg 775W (ATCC 43845) to ultrasonic waves under pressure treatments, at sublethal (manosonication) and lethal temperatures (manothermosonication) in citrate-phosphate buffer and in liquid whole egg was investigated. The influence of treatment parameters on the inactivation rate of manosonication was also studied. Decimal reduction times (Dt) of Salmonella Enteritidis, Salmonella Typhimurium, and Salmonella Senftenberg 775W corresponding to a heat treatment at 60 degrees C in pH 7 buffer and in liquid whole egg were 0.068, 0.12, and 1.0 min for buffer, and 0.12, 0.20, and 5.5 min for liquid whole egg. Those corresponding to a manosonication treatment (117 microns, 200 kPa, 40 degrees C) in both media were 0.73, 0.78, and 0.84 min, and 0.76, 0.84, and 1.4 min, respectively. When the amplitude of ultrasonic waves was increased linearly, the inactivation rate of manosonication increased exponentially. The inactivation rate also increased when pressure was raised. However, the magnitude of this increase was progressively smaller at higher pressures. The magnitude of the influence of the amplitude of ultrasonic waves and static pressure on the inactivation rate of manosonication was the same in the three serotypes investigated. Whereas a heat treatment at 60 degrees C only attained a 1/2-log cycle reduction in the number of Salmonella Senftenberg 775W survivors, a manothermosonication treatment (117 microns and 200 kPa) at this temperature attained a 3-log cycle reduction.

Effect of environmental factors and cell physiological state on Pulsed Electric Fields resistance and repair capacity of various strains of Escherichia coli.

The aim was to determine the resistance variation of four strains of Escherichia coli to Pulsed Electric Fields (PEF), the role of the sigma factor RpoS in PEF resistance, as well as the influence of several environmental factors and the cell physiological state on the PEF resistance and repair capacity. The rpoS null mutant, E. coli BJ4L1, exhibited decreased PEF resistance as compared with its wild-type parent, BJ4. W3110 and O157:H7 were the most PEF-resistant strains: whereas 2 and more than 3 Log10 cycles of BJ4 and BJ4L1 cells, respectively, were inactivated after 50 pulses at 35 kV/cm, only 0.5 Log10 cycle of inactivation of W3110 and O157:H7 was attained. A different pattern was observed and the resistance variation among strains was largely reduced, when selective recovery media were used. At exponential growth phase, the resistance of the four strains was lower, and more than 4 Log10 cycles of inactivation of all strains tested were attained at 30 kV/cm. Previous heat and cold shock treatments scarcely influenced cell PEF resistance. PEF survival increased with the reduction in water activity of the treatment medium to 0.94: the occurrence of sublethally injured cells was negligible, and less than 1 Log10 cycle of inactivation was attained at 35 kV/cm. PEF-treated cells were sensitive to a subsequent storage at pH 4.0 or in the presence of sorbic acid, attaining a final inactivation of 4-5 Log10 cycles after 24 hour-incubation. In conclusion, the work confirms the role of rpoS in PEF resistance. E. coli strains exhibit large differences in PEF resistance. These differences were less important when cells were recovered under selective conditions. Both resistance variation among strains and occurrence of sublethal damage were noticeably influenced by the environmental factors tested.

Inactivation of Salmonella enterica Serovar Enteritidis by Ultrasonic Waves under Pressure at Different Water Activities

ABSTRACT The inactivation of Salmonella enterica serovar Enteritidis by ultrasonic waves (20 kHz; 117-μm wavelength) under pressure (175 kPa) at nonlethal temperatures (manosonication [MS]) and lethal temperatures (manothermosonication [MTS]) in media of different water activities has been investigated. Heat decimal reduction time values increased 30 times when the water activity was decreased from nearly 1 to 0.96, but the MS resistance was increased only twofold. The inactivation of Salmonella serovar Enteritidis by ultrasound under pressure at low water activities was a phenomenon of the “all-or-nothing” type. A synergistic lethal effect was observed between heat and ultrasound in media with reduced water activity; the lower the water activity, the greater the synergistic effect. This work could be useful for improving sanitation and preservation treatments of foods, especially those which are sensitive to temperature and those in which components protect microorganisms to heat. It also contributes to our knowledge of microbial inactivation mechanisms by MS and MTS treatments.

Biosynthetic requirements for the repair of sublethal membrane damage in Escherichia coli cells after pulsed electric fields

Aims:  The aim was to evaluate the biosynthetic requirements for the repair of sublethal membrane damages in Escherichia coli cells after exposure to pulsed electric fields (PEF).

Variation in resistance of natural isolates of Staphylococcus aureus to heat, pulsed electric field and ultrasound under pressure

Aims:  To study and compare the resistance of 15 Staphylococcus aureus isolates to heat, pulsed electric field (PEF) and ultrasound (UW) under pressure (manosonication, MS).

Development of stress resistance in Staphylococcus aureus after exposure to sublethal environmental conditions.

The ability of Staphylococcus aureus to develop stress resistance responses was investigated. Exponential growth phase cells of S. aureus CECT 4459 were exposed to sublethal conditions (acid and alkaline pH, hydrogen peroxide, and heat) and then the acquisition of resistance to acid (pH 2.5), alkali (pH 12.0), hydrogen peroxide (50mM), and heat (58 degrees C) was determined. Conditions resulting in the maximum development of homologous resistance (tolerance to the same stress), while preventing lethal effects in the population, were pH 4.5 (2h), pH 9.5 (30 min), 0.05 mM H(2)O(2) (30 min), and 45 degrees C (2h). Under these adaptation conditions, times for the first decimal reduction (TFDC) to a lethal treatment at acid pH, alkaline pH, hydrogen peroxide, and heat were increased by a factor of 1.6, 2, 2, and 6, respectively. The presence of chloramphenicol or rifampicin in the adaptation medium completely abolished the increase in homologous resistance to acid pH and to hydrogen peroxide. By contrast, the development of homologous resistance to alkaline pH resulted independently of the presence of either chloramphenicol or rifampicin. S. aureus heat resistance increased in the presence of the inhibitors during the heat shock, but only partially. In some cases, the exposure to a given stress induced cross-protection against other agents. Protective combinations of sublethal stress and lethal agents were: acid pH-heat, acid pH-hydrogen peroxide, alkaline pH-hydrogen peroxide, heat-acid pH, and heat-hydrogen peroxide. These combinations of agents applied sequentially should be avoided in food-processing environments.

Predicting thermal inactivation in media of different pH of Salmonella grown at different temperatures

The influence of the growth temperature and the pH of the heating medium on the heat resistance at different temperatures of Salmonella typhimurium ATCC 13311 was studied and described mathematically. The shift of the growth temperature from 10 to 37 degrees C increased heat resistance of S. typhimurium fourfold. The pH of the heating medium at which heat resistance was maximum was pH 6 for cells grown at 37 degrees C, but changed with growth temperature. The alkalinization of the heating medium from pH 6 to pH 7.7 decreased the heat resistance of cells grown at 37 degrees C by a factor of 3. Neither the growth temperature nor the pH modified the z values significantly (4.9 degrees C). The decimal reduction times at different treatment temperatures, in buffers of different pH of cells of S. typhimurium grown at different temperatures, were accurately described by a mathematical equation (correlation coefficient of 0.97). This equation was also tested for Salmonella senftenberg 775W (ATCC 43845) and Salmonella enteritidis ATCC 13076, strains in which the correlation coefficients between the observed and the theoretically calculated values were 0.91 and 0.98, respectively.