Effects of pH and Acid Resistance on the Radiation Resistance of Enterohemorrhagic Escherichia coli

1999 ◽  
Vol 62 (3) ◽  
pp. 219-228 ◽  
Author(s):  
ROBERT L. BUCHANAN ◽  
SHARON G. EDELSON ◽  
GLENN BOYD
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Radiation Resistance ◽  
Brain Heart Infusion ◽  
Acid Resistance ◽  
Enterohemorrhagic Escherichia Coli ◽  
E Coli ◽  
Effects Of Ph ◽  
Ph Dependent ◽  
Subsequent Storage

The effects of pH and the induction of pH-dependent stationary-phase acid resistance on the radiation resistance of Escherichia coli were determined for seven enterohemorrhagic strains and one nonenterohemorrhagic strain. The isolates were grown in acidogenic or nonacidogenic media to pH levels of approximately 4.7 and 7.2, respectively. The cells were then transferred to brain heart infusion (BHI) broth adjusted to pH 4.0, 4.5, 5.0, and 5.5 (with HCl) that was preequilibrated to 2°C, and cultures were then irradiated using a 137Cs source. Surviving cells and the extent of injury were determined by plating on BHI and MacConkey agars both immediately after irradiation and after subsequent storage at 2°C for 7 days. Decreasing the pH of the BHI in which E. coli was irradiated had relatively little effect on the microorganism's radiation resistance. Substantial differences in radiation resistance were noted among strains, and induction of acid resistance consistently increased radiation resistance. Comparison of E. coli levels immediately after irradiation and after 7 days of refrigerated storage suggested that irradiation enhanced pH-mediated inactivation of the pathogen. These results demonstrate that prior growth under conditions that induce a pH-dependent stationary phase cross-protects E. coli against radiation inactivation and must be taken into account when determining the microorganism's irradiation D value.

Inactivation of Escherichia coli O157:H7 in Apple Juice by Irradiation

1998 ◽  
Vol 64 (11) ◽  
pp. 4533-4535 ◽  
Author(s):  
R. L. Buchanan ◽  
S. G. Edelson ◽  
K. Snipes ◽  
G. Boyd
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Radiation Resistance ◽  
Apple Juice ◽  
Suspended Solids ◽  
Acid Resistance ◽  
E Coli ◽  
Cesium 137 ◽  
Microbiological Criteria ◽  
Ph Dependent

ABSTRACT Three strains (932, Ent-C9490, and SEA13B88) of Escherichia coli O157:H7 were used to determine the effectiveness of low-dose gamma irradiation for eliminating E. coli O157:H7 from apple juice or cider and to characterize the effect of inducing pH-dependent, stationary-phase acid resistance on radiation resistance. The strains were grown in tryptic soy broth with or without 1% dextrose for 18 h to produce cells that were or were not induced to pH-dependent stationary-phase acid resistance. The bacteria were then transferred to clarified apple juice and irradiated at 2°C with a cesium-137 irradiator. Non-acid-adapted cells had radiationD values (radiation doses needed to decrease a microbial population by 90%) ranging from 0.12 to 0.21 kGy. D values increased to 0.22 to 0.31 kGy for acid-adapted cells. When acid-adapted SEA13B88 cells were tested in five apple juice brands having different levels of suspended solids (absorbances ranging from 0.04 to 2.01 at 550 nm), radiation resistance increased with increasing levels of suspended solids, with D values ranging from 0.26 to 0.35 kGy. Based on these results, a dose of 1.8 kGy should be sufficient to achieve the 5D inactivation of E. colirecommended by the National Advisory Committee for Microbiological Criteria for Foods.

pH-Dependent Stationary-Phase Acid Resistance Response of Enterohemorrhagic Escherichia coli in the Presence of Various Acidulants†

1999 ◽  
Vol 62 (3) ◽  
pp. 211-218 ◽  
Author(s):  
ROBERT L. BUCHANAN ◽  
SHARON G. EDELSON
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Stress Responses ◽  
Physiological Stress ◽  
Accurate Determination ◽  
Acid Resistance ◽  
Enterohemorrhagic Escherichia Coli ◽  
Resistance Response ◽  
E Coli ◽  
Ph Dependent

The effect of acidulant identity on the pH-dependent stationary-phase acid resistance response of enterohemorrhagic Escherichia coli was studied. Nine strains of E. coli (seven O157:H7, one O111:H-, and one biotype 1 reference strain) were cultured individually for 18 h at 37°C in tryptic soy broth (TSB) plus 1% dextrose and in TSB without dextrose to yield acid resistance induced and noninduced stationary-phase cells, respectively. These cultures were then inoculated into brain heart infusion broth (BHI) supplemented with 0.5% citric, malic, lactic, or acetic acid and adjusted to pH 3.0 with HCl. The BHI tubes were incubated at 37°C for up to 7 h and samples were removed after 0, 2, 5, and 7 h and plated for counting CFU on BHI agar and MacConkey agar (MA). The results were compared to data previously obtained with HCl only. Acid resistance varied substantially among the isolates, being dependent on the strain, the acidulant, and the induction of pH-dependent acid resistance. Hydrochloric acid was consistently the least damaging to cells; lactic acid was the most detrimental. The relative activity of the other acids was strain dependent. Inducing pH-dependent acid resistance increased the already substantial acid tolerance of stationary-phase E. coli. The extent of injury also varied with acid and strain, with as much as a 5-log-cycle differential between BHI agar and MA CFU counts. The accurate determination of the survival of enterohemorrhagic E. coli in acidic foods must take into account the biological variability of the microorganism with respect to its acid resistance and its ability to enhance survival through the induction of physiological stress responses.

scholarly journals Evolutionary Silence of the Acid Chaperone Protein HdeB in Enterohemorrhagic Escherichia coli O157:H7

2011 ◽  
Vol 78 (4) ◽  
pp. 1004-1014 ◽  
Author(s):  
Michelle Q. Carter ◽  
Jacqueline W. Louie ◽  
Clifton K. Fagerquist ◽  
Omar Sultan ◽  
William G. Miller ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Acid Resistance ◽  
Significant Loss ◽  
Enterohemorrhagic Escherichia Coli ◽  
Survival Strategies ◽  
Start Codon ◽  
Divergent Evolution ◽  
Content Type ◽  
E Coli ◽  
K 12

ABSTRACTThe periplasmic chaperones HdeA and HdeB are known to be important for cell survival at low pH (pH < 3) inEscherichia coliandShigellaspp. Here we investigated the roles of HdeA and HdeB in the survival of various enterohemorrhagicE. coli(EHEC) following exposure to pH 2.0. Similar to K-12 strains, the acid protections conferred by HdeA and HdeB in EHEC O145 were significant: loss of HdeA and HdeB led to over 100- to 1,000-fold reductions in acid survival, depending on the growth condition of prechallenge cells. However, this protection was much less inE. coliO157:H7 strains. Deletion ofhdeBdid not affect the acid survival of cells, and deletion ofhdeAled to less than a 5-fold decrease in survival. Sequence analysis of thehdeABoperon revealed a point mutation at the putative start codon of thehdeBgene in all 26E. coliO157:H7 strains analyzed, which shifted the ATG start codon to ATA. This mutation correlated with the lack of HdeB inE. coliO157:H7; however, the plasmid-borne O157-hdeBwas able to restore partially the acid resistance in anE. coliO145ΔhdeABmutant, suggesting the potential function of O157-HdeB as an acid chaperone. We conclude thatE. coliO157:H7 strains have evolved acid survival strategies independent of the HdeA/B chaperones and are more acid resistant than nonpathogenic K-12 for cells grown under nonfavorable culturing conditions such as in Luria-Bertani no-salt broth at 28°C. These results suggest a divergent evolution of acid resistance mechanisms withinE. coli.

Stationary-Phase Acid Resistance and Injury of Recent Bovine Escherichia coli O157 and non-O157 Biotype I Escherichia coli Isolates†

2004 ◽  
Vol 67 (3) ◽  
pp. 583-590 ◽  
Author(s):  
E. D. BERRY ◽  
G. A. BARKOCY-GALLAGHER ◽  
G. R. SIRAGUSA
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Acid Resistance ◽  
Low Ph ◽  
Acid Adaptation ◽  
E Coli ◽  
Log Reduction ◽  
Natural Isolates ◽  
Acidic Conditions ◽  
Ph Challenge

Stationary-phase acid resistance and the induction of acid resistance were assessed for recent bovine carcass isolates of Escherichia coli, including 39 serotype O157 strains and 20 non-O157 strains. When grown to stationary phase in the absence of glucose and without prior acid exposure, there was a range of responses to a pH challenge of 6 h at pH 2.5. However, populations of 53 of the 59 E. coli isolates examined were reduced by less than 2.00 log CFU/ml, and populations of 24 of these isolates were reduced by less than 1.00 log CFU/ml. In contrast, there was little variation in population reductions when the E. coli were grown with glucose and preadapted to acidic conditions. With few exceptions, acid adaptation improved survival to the acid challenge, with 57 of the 59 isolates exhibiting a log reduction of less than 0.50. Differences in acid resistance or the ability to adapt to acidic conditions between E. coli O157:H7 and non-O157 commensal E. coli were not observed. However, we did find that the E. coli O157 were disposed to greater acid injury after the low pH challenge than the non-O157 E. coli, both for cells that were and were not adapted to acidic conditions before the challenge. The enhancement of low pH survival after acid adaptation that was seen among these recent natural isolates of E. coli O157 further supports the idea that the previous environment of this pathogen should be a consideration when designing microbial safety strategies for foods preserved by low pH and acid.

Growth and survival of uninjured and sublethally heat-injured Escherichia coli O157:H7 on beef extract medium as influenced by package atmosphere and storage temperature

1999 ◽  
Vol 45 (3) ◽  
pp. 263-268 ◽  
Author(s):  
Jeffrey J Semanchek ◽  
David A Golden ◽  
Robert C Williams
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Storage Temperature ◽  
Brain Heart Infusion ◽  
Escherichia Coli O157 ◽  
Phase Growth ◽  
Growth And Survival ◽  
E Coli ◽  
Beef Extract ◽  
And Storage

The effect of atmospheric composition and storage temperature on growth and survival of uninjured and sublethally heat-injured Escherichia coli O157:H7, inoculated onto brain heart infusion agar containing 0.3% beef extract (BEM), was determined. BEM plates were packaged in barrier bags in air, 100% CO2, 100% N2, 20% CO2 : 80% N2, and vacuum and were stored at 4, 10, and 37°C for up to 20 days. Package atmosphere and inoculum status (i.e., uninjured or heat-injured) influenced (P < 0.01) growth and survival of E. coli O157:H7 stored at all test temperatures. Growth of heat-injured E. coli O157:H7 was slower (P < 0.01) than uninjured E. coli O157:H7 stored at 37°C. At 37°C, uninjured E. coli O157:H7 reached stationary phase growth earlier than heat-injured populations. Uninjured E. coli O157:H7 grew during 10 days of storage at 10°C, while heat-injured populations declined during 20 days of storage at 10°C. Uninjured E. coli O157:H7 stored at 10°C reached stationary phase growth within approximately 10 days in all packaging atmospheres except CO2. Populations of uninjured and heat-injured E. coli O157:H7 declined throughout storage for 20 days at 4°C. Survival of uninjured populations stored at 4°C, as well as heat-injured populations stored at 4 and 10°C, was enhanced in CO2 atmosphere. Survival of heat-injured E. coli O157:H7 at 4 and 10°C was not different (P > 0.05). Uninjured and heat-injured E. coli O157:H7 are able to survive at low temperatures in the modified atmospheres used in this study.Key words: E. coli O157:H7, sublethal injury, modified atmosphere packaging.

scholarly journals Distinct Acid Resistance and Survival Fitness Displayed by Curli Variants of Enterohemorrhagic Escherichia coli O157:H7

2011 ◽  
Vol 77 (11) ◽  
pp. 3685-3695 ◽  
Author(s):  
Michelle Q. Carter ◽  
Maria T. Brandl ◽  
Jacqueline W. Louie ◽  
Jennifer L. Kyle ◽  
Diana K. Carychao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Acid Resistance ◽  
Cell Aggregation ◽  
Oxygen Depletion ◽  
Growth Conditions ◽  
Enterohemorrhagic Escherichia Coli ◽  
Surface Attachment ◽  
Content Type ◽  
E Coli ◽  
Physiological Properties

ABSTRACTCurli are adhesive fimbriae ofEnterobacteriaceaeand are involved in surface attachment, cell aggregation, and biofilm formation. Here, we report that both inter- and intrastrain variations in curli production are widespread in enterohemorrhagicEscherichia coliO157:H7. The relative proportions of curli-producing variants (C+) and curli-deficient variants (C−) in anE. coliO157:H7 cell population varied depending on the growth conditions. In variants derived from the 2006 U.S. spinach outbreak strains, the shift between the C+and C−subpopulations occurred mostly in response to starvation and was unidirectional from C−to C+; in variants derived from the 1993 hamburger outbreak strains, the shift occurred primarily in response to oxygen depletion and was bidirectional. Furthermore, curli variants derived from the same strain displayed marked differences in survival fitness: C+variants grew to higher concentrations in nutrient-limited conditions than C−variants, whereas C−variants were significantly more acid resistant than C+variants. This difference in acid resistance does not appear to be linked to the curli fimbriaeper se, since acsgAdeletion mutant in either a C+or a C−variant exhibited an acid resistance similar to that of its parental strain. Our data suggest that natural curli variants ofE. coliO157:H7 carry several distinct physiological properties that are important for their environmental survival. Maintenance of curli variants in anE. coliO157:H7 population may provide a survival strategy in which C+variants are selected in a nutrient-limited environment, whereas C−variants are selected in an acidic environment, such as the stomach of an animal host, including that of a human.

Destruction of Escherichia coli O157:H7 and Salmonella typhimurium in Lebanon Bologna by Interaction of Fermentaron pH, Heating Temperature, and Time

1998 ◽  
Vol 61 (2) ◽  
pp. 152-157 ◽  
Author(s):  
KAMESWAR R. ELLAJOSYULA ◽  
STEPHANIE DOORES ◽  
EDWARD W. MILLS ◽  
RICHARD A. WILSON ◽  
RAMASWAMY C. ANANTHESWARAN ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Salmonella Typhimurium ◽  
Heating Temperature ◽  
Enterohemorrhagic Escherichia Coli ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Food Borne ◽  
Recent Outbreak ◽  
Food Borne Illness ◽  
Effects Of Ph

Fermented meats have caused food-borne illness due to enterohemorrhagic Escherichia coli. Consumption of Lebanon bologna was epidemiologically associated with a recent outbreak of salmonellosis. The present study was conducted to determine the effects of pH (after the fermentation step), final heating temperature, and time on destruction of E. coli O157:H7 and Salmonella typhimurium in Lebanon bologna. Raw Lebanon bologna mix was inoculated with either of the pathogens (ca. 108 CFU/g) and fermented for 12 h at 80°F (26.7°C) and then at 100°F (37.8°C) until the pH reached either 5.2 or 4.7. The mix was then heated to 110,115, or 120°F (43.3, 46.1, or 48.9°C). The bologna was sampled at various times, decimally diluted, and plated on either McConkey sorbitol agar or XLD agar to enumerate E. coli O157:H7 and S. typhimurium, respectively. Fermentation alone reduced populations of both pathogens by &lt;2 log units and heating alone reduced populations of E. coli O157:H7 by &lt; 3 log units. A combination of fermenting to either pH 5.2 or 4.7, followed by heating at 110°F (43.3°C) for 20 h, 115°F (46.1°C) for 10 h, or 120°F (48.9°C) for 3 h reduced populations of both pathogens by &gt;7 log units. Overall, S. typhimurium cells were either equally or significantly less resistant (P &lt; 0.01) than cells of E. coli O157:H7. Significant interactions (P &lt; 0.01) among the three factors for the destruction of E. coli O157:H7 were observed. A process-specific regression equation was developed to predict the destruction of E. coli O157:H7 in Lebanon bologna.

Effect of Prior Growth Conditions on the Thermal Inactivation of 13 Strains of Listeria monocytogenes in Two Heating Menstrua

2005 ◽  
Vol 68 (1) ◽  
pp. 168-172 ◽  
Author(s):  
SHARON G. EDELSON-MAMMEL ◽  
RICHARD C. WHITING ◽  
SAM W. JOSEPH ◽  
ROBERT L. BUCHANAN
Keyword(s):  
Listeria Monocytogenes ◽  
Thermal Resistance ◽  
Stationary Phase ◽  
Thermal Tolerance ◽  
Thermal Inactivation ◽  
Brain Heart Infusion ◽  
Acid Resistance ◽  
Growth Conditions ◽  
Ph Values ◽  
Ph Dependent

The thermal tolerance of 13 Listeria monocytogenes strains was tested using a submerged heating coil apparatus. The strains were grown individually for 18 h at 37°C in acidogenic tryptic soy broth (without dextrose) supplemented with 1% glucose and 1% glutamine (TSB+G) or nonacidogenic tryptic soy broth supplemented with 1% glutamine but containing no glucose (dextrose) (TSB−G). The former medium results in cells induced for pH-dependent, stationary-phase acid resistance, whereas the latter medium allows L. monocytogenes to grow to high numbers in the absence of glucose, yielding cells that are not induced for pH-dependent, stationary-phase acid resistance. The average final pH values of the 18-h TSB+G and the TSB−G cultures were 4.7 and 6.7, respectively. The cells grown in the acid resistance–inducing and non–acid resistance–inducing media were then tested in two heating menstrua that consisted of brain heart infusion broth adjusted to pH 3.0 and water activity (aw) of 0.987 or pH 7.0 and aw 0.970. In 14 of the 26 menstruum-strain combinations tested, the acid resistance–induced strains were more heat resistant then the equivalent noninduced cultures. No difference in the pattern of thermal resistance in response to induction of acid resistance was apparent among the different serovars tested. The results suggest that the ability of prior induction of acid resistance to enhance thermal resistance can vary substantially among L. monocytogenes strains.

scholarly journals Characterization of Enterohemorrhagic Escherichia coli Strains Based on Acid Resistance Phenotypes

2005 ◽  
Vol 73 (8) ◽  
pp. 4993-5003 ◽  
Author(s):  
Arvind A. Bhagwat ◽  
Lynn Chan ◽  
Rachel Han ◽  
Jasmine Tan ◽  
Mahendra Kothary ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Natural Populations ◽  
Acid Resistance ◽  
The United States ◽  
Growth Conditions ◽  
Enterohemorrhagic Escherichia Coli ◽  
Aerobic Growth ◽  
E Coli ◽  
Low Copy Number ◽  
Fermentative Growth

ABSTRACT Acid resistance is perceived to be an important property of enterohemorrhagic Escherichia coli strains, enabling the organisms to survive passage through the acidic environment of the stomach so that they may colonize the mammalian gastrointestinal tract and cause disease. Accordingly, the organism has developed at least three genetically and physiologically distinct acid resistance systems which provide different levels of protection. The glutamate-dependent acid resistance (GDAR) system utilizes extracellular glutamate to protect cells during extreme acid challenges and is believed to provide the highest protection from stomach acidity. In this study, the GDAR system of 82 pathogenic E. coli isolates from 34 countries and 23 states within the United States was examined. Twenty-nine isolates were found to be defective in inducing GDAR under aerobic growth conditions, while five other isolates were defective in GDAR under aerobic, as well as fermentative, growth conditions. We introduced rpoS on a low-copy-number plasmid into 26 isolates and were able to restore GDAR in 20 acid-sensitive isolates under aerobic growth conditions. Four isolates were found to be defective in the newly discovered LuxR-like regulator GadE (formerly YhiE). Defects in other isolates could be due to a mutation(s) in a gene(s) with an as yet undefined role in acid resistance since GadE and/or RpoS could not restore acid resistance. These results show that in addition to mutant alleles of rpoS, mutations in gadE exist in natural populations of pathogenic E. coli. Such mutations most likely alter the infectivity of individual isolates and may play a significant role in determining the infective dose of enterohemorrhagic E. coli.

scholarly journals Functional Heterogeneity of RpoS in Stress Tolerance of Enterohemorrhagic Escherichia coli Strains

2006 ◽  
Vol 72 (7) ◽  
pp. 4978-4986 ◽  
Author(s):  
Arvind A. Bhagwat ◽  
Jasmine Tan ◽  
Manan Sharma ◽  
Mahendra Kothary ◽  
Sharon Low ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Dna Sequences ◽  
Stress Responses ◽  
Glycogen Synthesis ◽  
Acid Tolerance ◽  
Acid Resistance ◽  
Growth Conditions ◽  
Enterohemorrhagic Escherichia Coli ◽  
Alkaline Stress ◽  
E Coli

ABSTRACT The stationary-phase sigma factor (RpoS) regulates many cellular responses to environmental stress conditions such as heat, acid, and alkali shocks. On the other hand, mutations at the rpoS locus have frequently been detected among pathogenic as well as commensal strains of Escherichia coli. The objective of this study was to perform a functional analysis of the RpoS-mediated stress responses of enterohemorrhagic E. coli strains from food-borne outbreaks. E. coli strains belonging to serotypes O157:H7, O111:H11, and O26:H11 exhibited polymorphisms for two phenotypes widely used to monitor rpoS mutations, heat tolerance and glycogen synthesis, as well as for two others, alkali tolerance and adherence to Caco-2 cells. However, these strains synthesized the oxidative acid resistance system through an rpoS-dependent pathway. During the transition from mildly acidic growth conditions (pH 5.5) to alkaline stress (pH 10.2), cell survival was dependent on rpoS functionality. Some strains were able to overcome negative regulation by RpoS and induced higher β-galactosidase activity without compromising their acid resistance. There were no major differences in the DNA sequences in the rpoS coding regions among the tested strains. The heterogeneity of rpoS-dependent phenotypes observed for stress-related phenotypes was also evident in the Caco-2 cell adherence assay. Wild-type O157:H7 strains with native rpoS were less adherent than rpoS-complemented counterpart strains, suggesting that rpoS functionality is needed. These results show that some pathogenic E. coli strains can maintain their acid tolerance capability while compromising other RpoS-dependent stress responses. Such adaptation processes may have significant impact on a pathogen's survival in food processing environments, as well in the host's stomach and intestine.

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