scholarly journals Inactivation of Escherichia coli andListeria innocua in Milk by Combined Treatment with High Hydrostatic Pressure and the Lactoperoxidase System

2000 ◽  
Vol 66 (10) ◽  
pp. 4173-4179 ◽  
Author(s):  
Cristina García-Graells ◽  
Caroline Valckx ◽  
Chris W. Michiels
Keyword(s):  
Escherichia Coli ◽  
High Pressure ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Combined Treatment ◽  
Food Preservation ◽  
Pressure Treatment ◽  
E Coli ◽  
Preservation Method ◽  
Lactoperoxidase System

ABSTRACT We have studied inactivation of four strains each ofEscherichia coli and Listeria innocua in milk by the combined use of high hydrostatic pressure and the lactoperoxidase-thiocyanate-hydrogen peroxide system as a potential mild food preservation method. The lactoperoxidase system alone exerted a bacteriostatic effect on both species for at least 24 h at room temperature, but none of the strains was inactivated. Upon high-pressure treatment in the presence of the lactoperoxidase system, different results were obtained for E. coli and L. innocua. For none of the E. coli strains did the lactoperoxidase system increase the inactivation compared to a treatment with high pressure alone. However, a strong synergistic interaction of both treatments was observed for L. innocua. Inactivation exceeding 7 decades was achieved for all strains with a mild treatment (400 MPa, 15 min, 20°C), which in the absence of the lactoperoxidase system caused only 2 to 5 decades of inactivation depending on the strain. Milk as a substrate was found to have a considerable effect protecting E. coli and L. innocua against pressure inactivation and reducing the effectiveness of the lactoperoxidase system under pressure on L. innocua. Time course experiments showed that L. innocua counts continued to decrease in the first hours after pressure treatment in the presence of the lactoperoxidase system.E. coli counts remained constant for at least 24 h, except after treatment at the highest pressure level (600 MPa, 15 min, 20°C), in which case, in the presence of the lactoperoxidase system, a transient decrease was observed, indicating sublethal injury rather than true inactivation.

scholarly journals Heat Shock Protein-Mediated Resistance to High Hydrostatic Pressure in Escherichia coli

2004 ◽  
Vol 70 (5) ◽  
pp. 2660-2666 ◽  
Author(s):  
Abram Aertsen ◽  
Kristof Vanoirbeek ◽  
Philipp De Spiegeleer ◽  
Jan Sermon ◽  
Kristel Hauben ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
High Pressure ◽  
Hydrostatic Pressure ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
High Hydrostatic Pressure ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
E Coli ◽  
Shock Proteins

ABSTRACT A random library of Escherichia coli MG1655 genomic fragments fused to a promoterless green fluorescent protein (GFP) gene was constructed and screened by differential fluorescence induction for promoters that are induced after exposure to a sublethal high hydrostatic pressure stress. This screening yielded three promoters of genes belonging to the heat shock regulon (dnaK, lon, clpPX), suggesting a role for heat shock proteins in protection against, and/or repair of, damage caused by high pressure. Several further observations provide additional support for this hypothesis: (i) the expression of rpoH, encoding the heat shock-specific sigma factor σ32, was also induced by high pressure; (ii) heat shock rendered E. coli significantly more resistant to subsequent high-pressure inactivation, and this heat shock-induced pressure resistance followed the same time course as the induction of heat shock genes; (iii) basal expression levels of GFP from heat shock promoters, and expression of several heat shock proteins as determined by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins extracted from pulse-labeled cells, was increased in three previously isolated pressure-resistant mutants of E. coli compared to wild-type levels.

scholarly journals Inactivation of Escherichia coli O157:H7 by High Hydrostatic Pressure Combined with Gas Packaging

2019 ◽  
Vol 7 (6) ◽  
pp. 154 ◽  
Author(s):  
Bing Zhou ◽  
Luyao Zhang ◽  
Xiao Wang ◽  
Peng Dong ◽  
Xiaosong Hu ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Escherichia Coli ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Cellular Structure ◽  
Combined Treatment ◽  
Physiological Saline ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Significant Difference

The inactivation of Escherichia coli O157:H7 (E. coli) in physiological saline and lotus roots by high hydrostatic pressure (HHP) in combination with CO2 or N2 was studied. Changes in the morphology, cellular structure, and membrane permeability of the cells in physiological saline after treatments were investigated using scanning electron microscopy, transmission electron microscopy, and flow cytometry, respectively. It was shown that after HHP treatments at 150–550 MPa, CO2-packed E. coli cells had higher inactivation than the N2-packed and vacuum-packed cells, and no significant difference was observed in the latter two groups. Further, both the morphology and intracellular structure of CO2-packed E.coli cells were strongly destroyed by high hydrostatic pressure. However, serious damage to the intracellular structures occurred in only the N2-packed E. coli cells. During HHP treatments, the presence of CO2 caused more disruptions in the membrane of E. coli cells than in the N2-packed and vacuum-packed cells. These results indicate that the combined treatment of HHP and CO2 had a strong synergistic bactericidal effect, whereas N2 did not have synergistic effects with HHP. Although these two combined treatments had different effects on the inactivation of E. coli cells, the inactivation mechanisms might be similar. During both treatments, E. coli cells were inactivated by cell damage induced to the cellular structure through the membrane components and the extracellular morphology, unlike the independent HHP treatment.

scholarly journals Induction of Oxidative Stress by High Hydrostatic Pressure in Escherichia coli

2005 ◽  
Vol 71 (5) ◽  
pp. 2226-2231 ◽  
Author(s):  
Abram Aertsen ◽  
Philipp De Spiegeleer ◽  
Kristof Vanoirbeek ◽  
Maria Lavilla ◽  
Chris W. Michiels
Keyword(s):  
Oxidative Stress ◽  
Escherichia Coli ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Applied Pressure ◽  
Pressure Treatment ◽  
E Coli ◽  
Cell Counts ◽  
Pressure Sensitive ◽  
High Hydrostatic Pressure Treatment

ABSTRACT Using leaderless alkaline phosphatase as a probe, it was demonstrated that pressure treatment induces endogenous intracellular oxidative stress in Escherichia coli MG1655. In stationary-phase cells, this oxidative stress increased with the applied pressure at least up to 400 MPa, which is well beyond the pressure at which the cells started to become inactivated (200 MPa). In exponential-phase cells, in contrast, oxidative stress increased with pressure treatment up to 150 MPa and then decreased again, together with the cell counts. Anaerobic incubation after pressure treatment significantly supported the recovery of MG1655, while mutants with increased intrinsic sensitivity toward oxidative stress (katE, katF, oxyR, sodAB, and soxS) were found to be more pressure sensitive than wild-type MG1655. Furthermore, mild pressure treatment strongly sensitized E. coli toward t-butylhydroperoxide and the superoxide generator plumbagin. Finally, previously described pressure-resistant mutants of E. coli MG1655 displayed enhanced resistance toward plumbagin. In one of these mutants, the induction of endogenous oxidative stress upon high hydrostatic pressure treatment was also investigated and found to be much lower than in MG1655. These results suggest that, at least under some conditions, the inactivation of E. coli by high hydrostatic pressure treatment is the consequence of a suicide mechanism involving the induction of an endogenous oxidative burst.

scholarly journals Induction of Shiga Toxin-Converting Prophage in Escherichia coli by High Hydrostatic Pressure

2005 ◽  
Vol 71 (3) ◽  
pp. 1155-1162 ◽  
Author(s):  
Abram Aertsen ◽  
David Faster ◽  
Chris W. Michiels
Keyword(s):  
Escherichia Coli ◽  
Hydrostatic Pressure ◽  
Shiga Toxin ◽  
High Hydrostatic Pressure ◽  
Sos Response ◽  
Resistant Mutant ◽  
Food Preservation ◽  
Bacterial Survival ◽  
E Coli ◽  
Pressure Resistance

ABSTRACT Since high hydrostatic pressure is becoming increasingly important in modern food preservation, its potential effects on microorganisms need to be thoroughly investigated. In this context, mild pressures (<200 MPa) have recently been shown to induce an SOS response in Escherichia coli MG1655. Due to this response, we observed a RecA- and LexA-dependent induction of lambda prophage upon treating E. coli lysogens with sublethal pressures. In this report, we extend this observation to lambdoid Shiga toxin (Stx)-converting bacteriophages in MG1655, which constitute an important virulence trait in Stx-producing E. coli strains (STEC). The window of pressures capable of inducing Stx phages correlated well with the window of bacterial survival. When pressure treatments were conducted in whole milk, which is known to promote bacterial survival, Stx phage induction could be observed at up to 250 MPa in E. coli MG1655 and at up to 300 MPa in a pressure-resistant mutant of this strain. In addition, we found that the intrinsic pressure resistance of two types of Stx phages was very different, with one type surviving relatively well treatments of up to 400 MPa for 15 min at 20°C. Interestingly, and in contrast to UV irradiation or mitomycin C treatment, pressure was not able to induce Stx prophage or an SOS response in several natural Stx-producing STEC isolates.

Study of the inactivation of Escherichia coli and pectin methylesterase in mango nectar under selected high hydrostatic pressure treatments

2011 ◽  
Vol 17 (6) ◽  
pp. 541-547 ◽  
Author(s):  
D. Bermúdez-Aguirre ◽  
J. Ángel Guerrero-Beltrán ◽  
G.V. Barbosa-Cánovas ◽  
J. Welti-Chanes
Keyword(s):  
Escherichia Coli ◽  
High Pressure ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Pectin Methylesterase ◽  
E Coli ◽  
Viable Cells ◽  
Plate Counts ◽  
The Media ◽  
Inactivation Effect

High hydrostatic pressure (HHP) was applied to fresh mango nectar (FMN) and sterilized mango nectar (SMN) to inactivate Escherichia coli and pectin methylesterase (PME). Pressure was applied at 275, 345 and 414 MPa. The come-up time (CUT) as well as 1, 2 and 4 min of treatment times were applied at the selected pressure to evaluate the inactivation effect on E. coli and PME. Total plate counts (TPC) were also evaluated in FMN. Results showed that mesophiles are inactivated in FMN to an important degree (up to 4 log) only with the CUT; the highest inactivation for mesophiles (7 log) was reported at 414 MPa after 4 min. Meanwhile, for E. coli 345 and 414 MPa after 2 and 1 min, respectively, were able to inactivate all viable cells in FMN. However, in SMN after 4 min at 275 MPa all cells of E. coli were also inactivated, showing the protective effect of the media between FMN and SMN. The PME showed its resistance to be inactivated with high pressure, showing the highest decrease in enzymatic activity (45%) after 4 min at 345 MPa but with an important activation at the highest pressure (414 MPa).

High-Pressure Transient Sensitization of Escherichia coli to Lysozyme and Nisin by Disruption of Outer-Membrane Permeability

1996 ◽  
Vol 59 (4) ◽  
pp. 350-355 ◽  
Author(s):  
KRISTEL J. A. HAUBEN ◽  
ELKE Y. WUYTACK ◽  
CARINE C. F. SOONTJENS ◽  
CHRIS W. MICHIELS
Keyword(s):  
Escherichia Coli ◽  
High Pressure ◽  
Outer Membrane ◽  
High Pressures ◽  
Ethylenediaminetetraacetic Acid ◽  
Viable Count ◽  
Water Soluble ◽  
Pressure Treatment ◽  
E Coli ◽  
Glucose Agar

Escherichia coli MG1655 suspensions in 10 mM phosphate buffer (pH 7.0) were subjected to high pressures in the range of 180 to 320 MPa for 15 min. Cell death was evident at 220 MPa and increased exponentially with pressure. Surviving populations were sublethally injured, as demonstrated by their reduced ability to form colonies on violet red bile glucose agar, a selective growth medium containing crystal violet and bile salts. During exposure to high pressure (&gt; 180 MPa), cells were sensitive to lysozyme, nisin, and ethylenediaminetetraacetic acid (EDTA), as was apparent from an increased lethality of pressure in the presence of these agents. Sublethal injury in the surviving population was lower in the presence of nisin and lysozyme, but higher in the presence of EDTA. Combinations of EDTA with nisin or lysozyme present during pressure treatment increased lethality in an additive manner. However, the addition of lysozyme, nisin and/or EDTA to pressurized cell suspensions immediately after pressure treatment did not cause any viable count reduction. Finally, we observed leakage of the periplasmic enzyme β-lactamase from an ampicillin-resistant recombinant E. coli MG1655 under high pressure. These results suggest that high pressure transiently disrupts the permeability of the E. coli outer membrane for water-soluble proteins.

Inactivation of Escherichia coli O157:H7 in Orange Juice Using a Combination of High Pressure and Mild Heat

1999 ◽  
Vol 62 (3) ◽  
pp. 277-279 ◽  
Author(s):  
M. LINTON ◽  
J. M. J. McCLEMENTS ◽  
M. F. PATTERSON
Keyword(s):  
Escherichia Coli ◽  
High Pressure ◽  
Resistant Strain ◽  
Orange Juice ◽  
Escherichia Coli O157 ◽  
Pressure Treatment ◽  
Processing Conditions ◽  
E Coli ◽  
Mild Heat ◽  
Ph Range

The effect of high pressure on the survival of a pressure-resistant strain of Escherichia coli O157:H7 (NCTC 12079) in orange juice was investigated over the pH range 3.4 to 5.0. The pH of commercial, sterile orange juice was adjusted to 3.4, 3.6, 3.9, 4.5, or 5.0. The juice was then inoculated with 108 CFU ml−1 of E. coli O157:H7. The inoculated orange juice was subjected to pressure treatments of 400, 500, or 550 MPa at 20°C or 30°C to determine the conditions that would give a 6-log10 inactivation of E. coli O157:H7. A pressure treatment of 550 MPa for 5 min at 20°C produced this level of kill at pH 3.4, 3.6, 3.9, and 4.5 but not at pH 5.0. Combining pressure treatment with mild heat (30°C) did result in a 6-log10 inactivation at pH 5.0. Thus, the processing conditions (temperature and time) must be considered when pressure-treating orange juice to ensure microbiological safety.

Combined Effect of High Hydrostatic Pressure and Nisin on Loss of Viability, Membrane Damage and Release of Intracellular Contents of Bacillus subtilis and Escherichia coli

2010 ◽  
Vol 6 (5) ◽  
Author(s):  
Wei-Min Qi ◽  
Ping Qian ◽  
Jian-Yong Yu ◽  
Chi-Yu Zhang ◽  
Xiao Chen ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Hydrostatic Pressure ◽  
Synergistic Effect ◽  
High Hydrostatic Pressure ◽  
Cell Envelope ◽  
Membrane Damage ◽  
Combined Effect ◽  
Gram Negative Bacteria ◽  
E Coli

Bacillus subtilis and Escherichia coli were chosen to investigate the combined effect of high hydrostatic pressure (HHP) and Nisin on loss of viability, membrane damage and release of intracellular contents of microorganisms. The results showed that the combination of 200 IU/mL Nisin and HHP exhibited a synergistic effect over 2 log on the inactivation of B. subtilis at pressure 300 MPa. The similar synergistic effect was observed on the membrane damage and release of intracellular contents of B. subtilis. The Nisin alone had no effect against E. coli, which belongs to gram negative bacteria. However, at pressure 300 MPa, Nisin caused the membrane damage from 55% to 80%. The synergistic effect of Nisin and HHP on loss of viability, membrane damage and release of intracellular contents of E. coli were also illustrated when the HHP pressure exceeded 300 MPa as the consequence of the serious changes produced by HHP at higher pressure in the cell envelope. It allows the entry of Nisin molecules to cell membrane.

Inactivation of Escherichia coli in Milk by High-Hydrostatic-Pressure Treatment in Combination with Antimicrobial Peptides

1999 ◽  
Vol 62 (11) ◽  
pp. 1248-1254 ◽  
Author(s):  
CRISTINA GARCÍA-GRAELLS ◽  
BARBARA MASSCHALCK ◽  
CHRIS W. MICHIELS
Keyword(s):  
Escherichia Coli ◽  
Hydrostatic Pressure ◽  
Antimicrobial Peptides ◽  
High Hydrostatic Pressure ◽  
Skim Milk ◽  
Substantial Improvement ◽  
Pressure Treatment ◽  
Different Temperatures ◽  
Resistant Mutants ◽  
High Concentrations

We studied the inactivation in milk of four Escherichia coli strains (MG1655 and three pressure-resistant mutants isolated from MG1655) by high hydrostatic pressure, alone or in combination with the natural antimicrobial peptides lysozyme and nisin and at different temperatures (10 to 50°C). Compared with that of phosphate buffer, the complex physicochemical environment of milk exerted a strong protective effect on E. coli MG1655 against high-hydrostatic-pressure inactivation, reducing inactivation from 7 logs at 400 MPa to only 3 logs at 700 MPa in 15 min at 20°C. An increase in lethality was achieved by addition of high concentrations of lysozyme (400 μg/ml) and nisin (400 IU/ml) to the milk before pressure treatment. The additional reduction amounted maximally to 3 logs in skim milk at 550 MPa but was strain dependent and significantly reduced in 1.55% fat and whole milk. An increase of the process temperature to 50°C also enhanced inactivation, particularly for the parental strain, but even in the presence of lysozyme and nisin, a 15-min treatment at 550 MPa and 50°C in skim milk allowed decimal reductions of only 4.5 to 6.9 for the pressure-resistant mutants. A substantial improvement of inactivation efficiency at ambient temperature was achieved by application of consecutive, short pressure treatments interrupted by brief decompressions. Interestingly, this pulsed-pressure treatment enhanced the sensitivity of the cells not only to high pressure but also to the action of lysozyme and nisin.

scholarly journals Synergetic Inactivation Mechanism of Protocatechuic Acid and High Hydrostatic Pressure against Escherichia coli O157:H7

Foods ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 3053
Author(s):  
Jingyi Hao ◽  
Yuqing Lei ◽  
Zhilin Gan ◽  
Wanbin Zhao ◽  
Junyan Shi ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Protocatechuic Acid ◽  
Combined Treatment ◽  
Membrane Integrity ◽  
Escherichia Coli O157 ◽  
Safety Issues ◽  
Bactericidal Effects ◽  
The Individual

With the wide application of high hydrostatic pressure (HHP) technology in the food industry, safety issues regarding food products, resulting in potential food safety hazards, have arisen. To address such problems, this study explored the synergetic bactericidal effects and mechanisms of protocatechuic acid (PCA) and HHP against Escherichia coli O157:H7. At greater than 200 MPa, PCA (1.25 mg/mL for 60 min) plus HHP treatments had significant synergetic bactericidal effects that positively correlated with pressure. After a combined treatment at 500 MPa for 5 min, an approximate 9.0 log CFU/mL colony decline occurred, whereas the individual HHP and PCA treatments caused 4.48 and 1.06 log CFU/mL colony decreases, respectively. Mechanistically, membrane integrity and morphology were damaged, and the permeability increased when E. coli O157: H7 was exposed to the synergetic stress of PCA plus HHP. Inside cells, the synergetic treatment additionally targeted the activities of enzymes such as superoxide dismutase, catalase and ATPase, which were inhibited significantly (p ≤ 0.05) when exposed to high pressure. Moreover, an analysis of circular dichroism spectra indicated that the synergetic treatment caused a change in DNA structure, which was expressed as the redshift of the characteristic absorption peak. Thus, the synergetic treatment of PCA plus HHP may be used as a decontamination method owing to the good bactericidal effects on multiple targets.

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