Augmentation of Killing of Escherichia coli O157 by Combinations of Lactate, Ethanol, and Low-pH Conditions

1999 ◽  
Vol 65 (3) ◽  
pp. 1308-1311 ◽  
Author(s):  
Sarah L. Jordan ◽  
Jayne Glover ◽  
Laura Malcolm ◽  
Fiona M. Thomson-Carter ◽  
Ian R. Booth ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Acid Tolerance ◽  
Low Ph ◽  
Exponential Phase ◽  
Escherichia Coli O157 ◽  
Cytoplasmic Ph ◽  
Ph Homeostasis ◽  
Acid Tolerant ◽  
Ph Conditions

ABSTRACT The acid tolerance of Escherichia coli O157:H7 strains can be overcome by addition of lactate, ethanol, or a combination of the two agents. Killing can be increased by as much as 4 log units in the first 5 min of incubation at pH 3 even for the most acid-tolerant isolates. Exponential-phase, habituated, and stationary-phase cells are all sensitive to incubation with lactate and ethanol. Killing correlates with disruption of the capacity for pH homeostasis. Habituated and stationary-phase cells can partially offset the effects of the lowering of cytoplasmic pH.

Survival of Low-pH Stress by Escherichia coli O157:H7: Correlation between Alterations in the Cell Envelope and Increased Acid Tolerance

1999 ◽  
Vol 65 (7) ◽  
pp. 3048-3055 ◽  
Author(s):  
Kieran N. Jordan ◽  
Lynn Oxford ◽  
Conor P. O’Byrne
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Cell Envelope ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Composition ◽  
Acid Tolerance ◽  
Low Ph ◽  
Exponential Phase ◽  
Viability Loss ◽  
High Level

ABSTRACT Survival of a nontoxigenic isolate of Escherichia coliO157:H7 at low pH (pH 3.0) was examined over prolonged time periods for each of three population types: exponential-phase cells, stationary-phase cells, and acid-adapted exponential-phase cells. In each population, approximately 5 × 104 CFU ml−1 were detected after a 24-h incubation at pH 3.0. Even after 3 days at pH 3.0, significant numbers of survivors from each of the three populations could be detected. The high level of acid tolerance exhibited by these survivors was found to be quickly lost once they were transferred to conditions which permitted growth to resume, indicating that they were not mutants. Proton flux measurements on the three populations of cells revealed that the initial rates of viability loss at pH 3.0 correlated well with net proton accumulation. Cells showing a high initial rate of viability loss (exponential-phase cells) accumulated protons at the highest rate, whereas resistant populations (adapted or stationary-phase cells) accumulated protons only slowly. Differences in the protein composition of the cell envelope between the three populations were studied by two-dimensional polyacrylamide gel electrophoresis. Complex differences in the pattern of proteins expressed by each population were uncovered. The implications of these findings are discussed in the context of a possible model accounting for acid tolerance in this important food-borne pathogen.

scholarly journals Enhanced Acid Sensitivity of Pressure-Damaged Escherichia coli O157 Cells

2001 ◽  
Vol 67 (4) ◽  
pp. 1983-1985 ◽  
Author(s):  
Rafael Pagán ◽  
Sarah Jordan ◽  
Amparo Benito ◽  
Bernard Mackey
Keyword(s):  
Escherichia Coli ◽  
Escherichia Coli O157 ◽  
Cytoplasmic Ph ◽  
Ph Homeostasis ◽  
Acid Sensitivity ◽  
Acid Labile

ABSTRACT Pressure-damaged Escherichia coli O157 cells were more acid sensitive than native cells and were impaired in pH homeostasis. However differences in acid sensitivity were not related to differences in cytoplasmic pH (pHi). Cellular β-galactosidase was more acid labile in damaged cells. Sensitization to acid may thus involve loss of protective or repair functions.

scholarly journals Acid tolerance in early colonizers of oral biofilms

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Gabriella Boisen ◽  
Julia R. Davies ◽  
Jessica Neilands
Keyword(s):  
Acid Tolerance ◽  
Low Ph ◽  
Salivary Proteins ◽  
Confocal Scanning Laser Microscopy ◽  
Planktonic Cells ◽  
Tolerance Development ◽  
Salivary Pellicle ◽  
Oral Biofilms ◽  
Acid Tolerant

Abstract Background In caries, low pH drives selection and enrichment of acidogenic and aciduric bacteria in oral biofilms, and development of acid tolerance in early colonizers is thought to play a key role in this shift. Since previous studies have focussed on planktonic cells, the effect of biofilm growth as well as the role of a salivary pellicle on this process is largely unknown. We explored acid tolerance and acid tolerance response (ATR) induction in biofilm cells of both clinical and laboratory strains of three oral streptococcal species (Streptococcus gordonii, Streptococcus oralis and Streptococcus mutans) as well as two oral species of Actinomyces (A. naeslundii and A. odontolyticus) and examined the role of salivary proteins in acid tolerance development. Methods Biofilms were formed on surfaces in Ibidi® mini flow cells with or without a coating of salivary proteins and acid tolerance assessed by exposing them to a challenge known to kill non-acid tolerant cells (pH 3.5 for 30 min) followed by staining with LIVE/DEAD BacLight and confocal scanning laser microscopy. The ability to induce an ATR was assessed by exposing the biofilms to an adaptation pH (pH 5.5) for 2 hours prior to the low pH challenge. Results Biofilm formation significantly increased acid tolerance in all the clinical streptococcal strains (P < 0.05) whereas the laboratory strains varied in their response. In biofilms, S. oralis was much more acid tolerant than S. gordonii or S. mutans. A. naeslundii showed a significant increase in acid tolerance in biofilms compared to planktonic cells (P < 0.001) which was not seen for A. odontolyticus. All strains except S. oralis induced an ATR after pre-exposure to pH 5.5 (P < 0.05). The presence of a salivary pellicle enhanced both acid tolerance development and ATR induction in S. gordonii biofilms (P < 0.05) but did not affect the other bacteria to the same extent. Conclusions These findings suggest that factors such as surface contact, the presence of a salivary pellicle and sensing of environmental pH can contribute to the development of high levels of acid tolerance amongst early colonizers in oral biofilms which may be important in the initiation of caries.

Mannose-Binding Activity of Escherichia coli: a Determinant of Attachment and Ingestion of the Bacteria by Macrophages

1980 ◽  
Vol 29 (2) ◽  
pp. 417-424
Author(s):  
Zvi Bar-Shavit ◽  
Rachel Goldman ◽  
Itzhak Ofek ◽  
Nathan Sharon ◽  
David Mirelman
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Pathogenic Bacteria ◽  
Binding Activity ◽  
Exponential Phase ◽  
Restrictive Temperature ◽  
Filamentous Bacteria ◽  
Phagocytic Cells ◽  
E Coli ◽  
Mannose Binding

Recently, it was suggested that a mannose-specific lectin on the bacterial cell surface is responsible for the recognition by phagocytic cells of certain nonopsonized Escherichia coli strains. In this study we assessed the interaction of two strains of E. coli at different phases of growth with a monolayer of mouse peritoneal macrophages and developed a direct method with [ 14 C]mannan to quantitate the bacterial mannose-binding activity. Normal-sized bacteria were obtained from logarithmic and stationary phases of growth. Nonseptated filamentous cells were formed by growing the organisms in the presence of cephalexin or at a restrictive temperature. Attachment to macrophages of all bacterial forms was inhibited by methyl α- d -mannoside and mannan but not by other sugars tested. The attachment of stationary phase and filamentous bacteria to macrophages, as well as their mannose-binding activity, was similar, whereas in the exponential-phase bacteria they were markedly reduced. The results show a linear relation between the two parameters ( R = 0.98, P < 0.001). The internalization of the filamentous cells attached to macrophages during 45 min of incubation was much less efficient (20%) compared to that of exponential-phase, stationary-phase, or antibody-coated filamentous bacteria (90%). The results indicate that the mannose-binding activity of E. coli determines the recognition of the organisms by phagocytes. They further suggest that administration of β-lactam antibiotics may impair elimination of certain pathogenic bacteria by inducing the formation of filaments which are inefficiently internalized by the host's phagocytic cells.

scholarly journals Contribution of dps to Acid Stress Tolerance and Oxidative Stress Tolerance in Escherichia coli O157:H7

2000 ◽  
Vol 66 (9) ◽  
pp. 3911-3916 ◽  
Author(s):  
Sang Ho Choi ◽  
David J. Baumler ◽  
Charles W. Kaspar
Keyword(s):  
Oxidative Stress ◽  
Escherichia Coli ◽  
Hydrogen Peroxide ◽  
Stationary Phase ◽  
Stress Tolerance ◽  
Parent Strain ◽  
Acid Stress ◽  
Acid Tolerance ◽  
Gastric Fluid ◽  
Acid Challenge

ABSTRACT An Escherichia coli O157:H7dps::nptI mutant (FRIK 47991) was generated, and its survival was compared to that of the parent in HCl (synthetic gastric fluid, pH 1.8) and hydrogen peroxide (15 mM) challenges. The survival of the mutant in log phase (5-h culture) was significantly impaired (4-log10-CFU/ml reduction) compared to that of the parent strain (ca. 1.0-log10-CFU/ml reduction) after a standard 3-h acid challenge. Early-stationary-phase cells (12-h culture) of the mutant decreased by ca. 4 log10CFU/ml while the parent strain decreased by approximately 2 log10 CFU/ml. No significant differences in the survival of late-stationary-phase cells (24-h culture) between the parent strain and the mutant were observed, although numbers of the parent strain declined less in the initial 1 h of acid challenge. FRIK 47991 was more sensitive to hydrogen peroxide challenge than was the parent strain, although survival improved in stationary phase. Complementation of the mutant with a functional dps gene restored acid and hydrogen peroxide tolerance to levels equal to or greater than those exhibited by the parent strain. These results demonstrate that decreases in survival were from the absence of Dps or a protein regulated by Dps. The results from this study establish that Dps contributes to acid tolerance in E. coli O157:H7 and confirm the importance of Dps in oxidative stress protection.

Viability of Acid-Adapted Escherichia coli O157:H7 in Ground Beef Treated with Acidic Calcium Sulfate

2004 ◽  
Vol 67 (3) ◽  
pp. 591-595 ◽  
Author(s):  
LARRY R. BEUCHAT ◽  
ALAN J. SCOUTEN
Keyword(s):  
Escherichia Coli ◽  
Acetic Acid ◽  
Calcium Sulfate ◽  
Ground Beef ◽  
Storage Period ◽  
Acid Tolerance ◽  
Escherichia Coli O157 ◽  
E Coli ◽  
Acidic Environments ◽  
Subsequent Acid

The effects of lactic acid, acetic acid, and acidic calcium sulfate (ACS) on viability and subsequent acid tolerance of three strains of Escherichia coli O157:H7 were determined. Differences in tolerance to acidic environments were observed among strains, but the level of tolerance was not affected by the acidulant to which cells had been exposed. Cells of E. coli O157:H7 adapted to grow on tryptic soy agar acidified to pH 4.5 with ACS were compared to cells grown at pH 7.2 in the absence of ACS for their ability to survive after inoculation into ground beef treated with ACS, as well as untreated beef. The number of ACS-adapted cells recovered from ACS-treated beef was significantly (α = 0.05) higher than the number of control cells recovered from ACS-treated beef during the first 3 days of a 10-day storage period at 4°C, suggesting that ACS-adapted cells might be initially more tolerant than unadapted cells to reduced pH in ACS-treated beef. Regardless of treatment of ground beef with ACS or adaptation of E. coli O157:H7 to ACS before inoculating ground beef, the pathogen survived in high numbers.

scholarly journals Studies on deoxyribonucleic acid-dependent ribonucleic acid polymerase from Escherichia coli. Variations of the enzyme activity during growth

1972 ◽  
Vol 129 (2) ◽  
pp. 291-299 ◽  
Author(s):  
K. A. Abraham ◽  
K. J. Andersen ◽  
A. Rognes
Keyword(s):  
Escherichia Coli ◽  
Stationary Phase ◽  
Stationary Phases ◽  
Bacteriophage T4 ◽  
Polymerase Activity ◽  
Exponential Phase ◽  
Enzyme Preparations ◽  
Rna Polymerase Activity ◽  
Enzyme Molecules ◽  
Cellular Dna

1. RNA polymerase activity of Escherichia coli extracts prepared from cells in exponential and stationary phases of growth, when measured in the presence and absence of external template, showed significant qualitative differences. 2. In both extracts, polymerase activity was higher when assayed with external template, suggesting the presence of a pool of enzyme not bound to cellular DNA. 3. In the crude extract, the fraction of enzyme bound to cellular DNA is higher during the exponential phase of growth. 4. A method is described for the purification of enzyme molecules not tightly bound to cellular DNA from exponential- and stationary-phase cultures. 5. Purified enzyme preparations showed differences in template requirement and subunit composition. 6. On phosphocellulose chromatography of stationary-phase enzyme, a major portion of polymerase activity eluted from the column with 0.25m-KCl. In the case of exponential-phase enzyme, polymerase activity eluted from a phosphocellulose column mainly with 0.35m-KCl. 7. Enzyme assays done with excess of bacteriophage T4 DNA showed a strong inhibition of stationary-phase enzyme by this template. The exponential-phase enzyme was only slightly inhibited by excess of bacteriophage T4 DNA.

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