Fate of Escherichia coli Strains Inoculated in Model Cheese Elaborated with or without Starter and Treated by High Hydrostatic Pressure

2006 ◽  
Vol 69 (12) ◽  
pp. 2856-2864 ◽  
Author(s):  
SÍLVIA DE LAMO-CASTELLVÍ ◽  
MARTA CAPELLAS ◽  
ARTUR X. ROIG-SAGUÉS ◽  
TOMÁS LÓPEZ-PEDEMONTE ◽  
M. MANUELA HERNÁNDEZ-HERRERO ◽  
...  

The aim of this research was to study high hydrostatic pressure inactivation of two strains of Escherichia coli (E. coli O59:H21 [CECT 405] and E. coli O157:H7 [CECT 5947]) inoculated in washed-curd model cheese elaborated with and without starter and the ability of these strains for survival, recovery, and growth. Samples were treated at 300, 400, and 500 MPa for 10 min at 20°C and analyzed after the treatment and after 1, 7, and 15 days of storage at 8°C to study the behavior of Escherichia populations. Cheeses elaborated with starter showed the maximum lethality at 400 and 500 MPa, and no significant differences in the baroresistant behavior of either strains were detected, except for E. coli O157:H7 at 400 MPa in cell counts obtained with thin agar layer method medium, where the decrease value was significantly lower. In cheese elaborated without starter, the highest decrease value was observed at 500 MPa, except for E. coli O59:H21 in cell counts obtained with selective culture medium, where the highest decrease value was also found at 400 MPa. The ability to repair and grow was not observed in model cheese elaborated with starter, as cell counts of treated samples decreased after 15 days of storage at 8°C. By contrast, in cheese elaborated without starter, all pressurized samples showed the trend to repair and grow during the storage period in both strains. These results suggest that the presence of starter and low pH values are the main factors that control the ability of Escherichia strains inoculated in this type of cheese and treated by high hydrostatic pressure to recover and grow.

2005 ◽  
Vol 71 (5) ◽  
pp. 2226-2231 ◽  
Author(s):  
Abram Aertsen ◽  
Philipp De Spiegeleer ◽  
Kristof Vanoirbeek ◽  
Maria Lavilla ◽  
Chris W. Michiels

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.


2004 ◽  
Vol 70 (5) ◽  
pp. 2660-2666 ◽  
Author(s):  
Abram Aertsen ◽  
Kristof Vanoirbeek ◽  
Philipp De Spiegeleer ◽  
Jan Sermon ◽  
Kristel Hauben ◽  
...  

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.


Author(s):  
Wei-Min Qi ◽  
Ping Qian ◽  
Jian-Yong Yu ◽  
Chi-Yu Zhang ◽  
Xiao Chen ◽  
...  

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.


2000 ◽  
Vol 63 (12) ◽  
pp. 1713-1718 ◽  
Author(s):  
ERROL V. RAGHUBEER ◽  
C. PATRICK DUNNE ◽  
DANIEL F. FARKAS ◽  
EDMUND Y. TING

The effects of high hydrostatic pressure (HPP; 545 MPa) on strains of Escherichia coli O157:H7, Listeria monocytogenes, enterotoxigenic Staphylococcus aureus, and nonpathogenic microorganisms were studied in tomato-based salsa. Products were evaluated for the survival of the inoculated pathogens following HPP treatment and after storage at 4°C and 21 to 23°C for up to 2 months. Inoculated samples without HPP treatment, stored under the same conditions, were also evaluated to determine the effects of the acid environment of salsa on the survival of inoculated strains. None of the inoculated pathogens were detected in the HPP-treated samples for all treatments throughout the storage period. Inoculated pathogens were detected in the non–HPP-treated samples stored at 4°C after 1 month, with L. monocytogenes showing the highest level of survivors. In the non–HPP-treated samples stored at 21 to 23°C, E. coli and S. aureus were not detected after 1 week, but L. monocytogenes was detected in low levels. Studies with nonpathogenic strains of the pathogens were conducted at Oregon State University using HPP treatments in a semicontinuous production system. The nonpathogenic microorganisms (E. coli, Listeria innocua, Listeria welshimeri, and nonenterotoxigenic S. aureus) were inoculated together into a feeder tank containing 100 liters of salsa. Microbiological results of samples collected before HPP treatment and from the aseptic filler were similar to those obtained for the pathogenic strains. No survivors were detected in any of the HPP-treated samples.


2000 ◽  
Vol 66 (10) ◽  
pp. 4173-4179 ◽  
Author(s):  
Cristina García-Graells ◽  
Caroline Valckx ◽  
Chris W. Michiels

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.


2001 ◽  
Vol 67 (10) ◽  
pp. 4901-4907 ◽  
Author(s):  
Marianne Robey ◽  
Amparo Benito ◽  
Roger H. Hutson ◽  
Cristina Pascual ◽  
Simon F. Park ◽  
...  

ABSTRACT Several natural isolates of Escherichia coliO157:H7 have previously been shown to exhibit stationary-phase-dependent variation in their resistance to inactivation by high hydrostatic pressure. In this report we demonstrate that loss of the stationary-phase-inducible sigma factor RpoS resulted in decreased resistance to pressure inE. coli O157:H7 and in a commensal strain. Furthermore, variation in the RpoS activity of the natural isolates of O157:H7 correlated with the pressure resistance of those strains. Heterogeneity was noted in the rpoS alleles of the natural isolates that may explain the differences in RpoS activity. These results are consistent with a role for rpoS in mediating resistance to high hydrostatic pressure in E. coliO157:H7.


2003 ◽  
Vol 9 (6) ◽  
pp. 403-407 ◽  
Author(s):  
C. Dogan ◽  
O. Erkmen

Inactivation of Escherichia coli by high hydrostatic pressure (UHHP) was determined in broth, milk and orange and peach juices inoculated with the bacteria. HHP ranged from 200 to 700 MPa at 25 C and different treatment times. No cell growth occurred in broth after 60, 25, 15, 10 and 7 min at 300, 400, 500, 600 and 700 MPa, respectively. Reduction of aerobic bacteria in milk and peach juice were 3.08 and 6.07 log units after 15 min at 400 MPa, respectively, while all bacterial cells were inactivated in orange juice. Sterilisation of raw milk contaminated with E. coli occurred at 600 MPa for 30 min, while peach and orange juices needed 12 and 10 min, respectively. The injury of cells in broth at 300 MPa ranged from 8.8 to 100% depending on magnitude of pressure and treated time. In general, inactivation of aerobic bacteria and E. coli was enhanced significantly (P<0.01) by increasing the pressure.


2014 ◽  
Vol 77 (5) ◽  
pp. 781-787 ◽  
Author(s):  
NICOLAS A. LAVIERI ◽  
JOSEPH G. SEBRANEK ◽  
JOSEPH C. CORDRAY ◽  
JAMES S. DICKSON ◽  
ASHLEY M. HORSCH ◽  
...  

Sodium nitrite exerts an inhibitory effect on the growth of Listeria monocytogenes. The objective of this study was to investigate the effects of various nitrite concentrations from a vegetable source with and without high hydrostatic pressure (HHP) on the recovery and growth of L. monocytogenes on ready-to-eat restructured ham. A preconverted celery powder was used as the vegetable source of nitrite. Targeted concentrations of natural nitrite investigated were 0, 50, and 100 mg/kg. HHP treatments evaluated were 400 MPa for 4 min and 600 MPa for 1 or 4 min at 12 ± 2°C (initial temperature of the pressurization fluid). Viable L. monocytogenes populations were monitored on modified Oxford medium and thin agar layer medium through 98 days of storage at 4 ± 1°C. Populations on both media did not differ. The HHP treatment at 600 MPa for 4 min resulted in L. monocytogenes populations below the detection limit of our sampling protocols throughout the storage period regardless of the natural nitrite concentration. The combination of HHP at 400 MPa for 4 min or 600 MPa for 1 min with natural nitrite resulted in initial inhibition of viable L. monocytogenes. Ham formulations that did not contain natural nitrite allowed faster growth of L. monocytogenes than did those with nitrite, regardless of whether they were treated with HHP. The results indicate that nitrite from a vegetable source at the concentrations used in this study resulted in slower growth of this microorganism. HHP treatments enhanced the inhibitory effects of natural nitrite on L. monocytogenes growth. Thus, the combination of natural nitrite plus HHP appears to have a synergistic inhibitory effect on L. monocytogenes growth.


2016 ◽  
Vol 82 (22) ◽  
pp. 6656-6663 ◽  
Author(s):  
Elisa Gayán ◽  
Alexander Cambré ◽  
Chris W. Michiels ◽  
Abram Aertsen

ABSTRACTThe development of resistance in foodborne pathogens to food preservation techniques is an issue of increasing concern, especially in minimally processed foods where safety relies on hurdle technology. In this context, mild heat can be used in combination with so-called nonthermal processes, such as high hydrostatic pressure (HHP), at lower individual intensities to better retain the quality of the food. However, mild stresses may increase the risk of (cross-)resistance development in the surviving population, which in turn might compromise food safety. In this investigation, we examined the evolution ofEscherichia coliO157:H7 strain ATCC 43888 after recurrent exposure to progressively intensifying mild heat shocks (from 54.0°C to 60.0°C in 0.5°C increments) with intermittent resuscitation and growth of survivors. As such, mutant strains were obtained after 10 cycles of selection with ca. 106-fold higher heat resistance than that for the parental strain at 58.0°C, although this resistance did not extend to temperatures exceeding 60.0°C. Moreover, these mutant strains typically displayed cross-resistance against HHP shock and displayed signs of enhanced RpoS and RpoH activity. Interestingly, additional cycles of selection maintaining the intensity of the heat shock constant (58.5°C) selected for mutant strains in which resuscitation speed, rather than resistance, appeared to be increased. Therefore, it seems that resistance and resuscitation speed are rapidly evolvable traits inE. coliATCC 43888 that can compromise food safety.IMPORTANCEIn this investigation, we demonstrated thatEscherichia coliO157:H7 ATCC 43888 rapidly acquires resistance to mild heat exposure, with this resistance yielding cross-protection to high hydrostatic pressure treatment. In addition, mutants ofE. coliATCC 43888 in which resuscitation speed, rather than resistance, appeared to be improved were selected. As such, both resistance and resuscitation speed seem to be rapidly evolvable traits that can compromise the control of foodborne pathogens in minimal processing strategies, which rely on the efficacy of combined mild preservation stresses for food safety.


2006 ◽  
Vol 69 (3) ◽  
pp. 542-547 ◽  
Author(s):  
KWAN DEOG MOON ◽  
PASCAL DELAQUIS ◽  
PETER TOIVONEN ◽  
SUSAN BACH ◽  
KAREEN STANICH ◽  
...  

The behavior of Escherichia coli O157:H7 in Granny Smith, Gala, Empire, McIntosh, Red Delicious, and Golden Delicious apple juice with or without supplementation with 5 or 10 mM vanillic acid was examined over a storage period of 7 days at 4 and 15°C. The consequences of supplementation on sensory difference and preference were also determined by triangle testing. Juices made from the six apple cultivars had pH values ranging between pH 3.13 and 3.92. Vanillic acid exerted a concentration, pH, and time-dependent lethal effect toward E. coli O157:H7 in unpasteurized apple juice. Supplementation with 10 mM vanillic acid led to a 5-logarithm reduction in populations after 7 days at both temperatures, but sensory analysis revealed significant differences from and preference for unsupplemented juices. Supplementation with 5 mM vanillic acid accelerated death of E. coli O157:H7, but population reductions ranged from 5 log CFU/ml in low pH juices to none in high pH juices, particularly at 4°C. No sensory difference or preference was detected in two of the six juices at this level of supplementation.


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