scholarly journals Impact of Inoculum Preparation and Storage Conditions on the Response of Escherichia coli O157:H7 Populations to Undercooking and Simulated Exposure to Gastric Fluid

2006 ◽  
Vol 72 (1) ◽  
pp. 672-679 ◽  
Author(s):  
Jarret D. Stopforth ◽  
Panagiotis N. Skandamis ◽  
Laura V. Ashton ◽  
Ifigenia Geornaras ◽  
Patricia A. Kendall ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Acid Tolerance ◽  
Storage Conditions ◽  
Gastric Fluid ◽  
Simulated Gastric Fluid ◽  
Tissue Samples ◽  
E Coli ◽  
Inoculum Preparation ◽  
The Impact ◽  
And Storage

ABSTRACT This study evaluated the impact of inoculum preparation and storage conditions on the response of Escherichia coli O157:H7 exposed to consumer-induced stresses simulating undercooking and digestion. Lean beef tissue samples were inoculated with E. coli O157:H7 cultures prepared in tryptic soy broth or meat decontamination runoff fluids (WASH) or detached from moist biofilms or dried biofilms formed on stainless steel coupons immersed in inoculated WASH. After inoculation, the samples were left untreated or dipped for 30 s each in hot (75°C) water followed by lactic acid (2%, 55°C), vacuum packaged, stored at 4 (28 days) or 12°C (16 days), and periodically transferred to aerobic storage (7°C for 5 days). During storage, samples were exposed to sequential heat (55°C; 20 min) and simulated gastric fluid (adjusted to pH 1.0 with HCl; 90 min) stresses simulating consumption of undercooked beef. Under the conditions of this study, cells originating from inocula of planktonic cells were, in general, more resistant to heat and acid than cells from cultures grown as biofilms and detached prior to meat inoculation. Heat and acid tolerance of cells on meat stored at 4°C was lower than that of cells on nondecontaminated meat stored at 12°C, where growth occurred during storage. Decontamination of fresh beef resulted in injury that inhibited subsequent growth of surviving cells at 12°C, as well as in decreases in resistance to subsequent heat and acid stresses. The shift of pathogen cells on beef stored under vacuum at 4°C to aerobic storage did not affect cell populations or subsequent survival after sequential exposure to heat and simulated gastric fluid. However, the transfer of meat stored under vacuum at 12°C to aerobic storage resulted in reduction in pathogen counts during aerobic storage and sensitization of survivors to the effects of sequential heat and acid exposure.

scholarly journals Investigation of multidrug-resistant fatal colisepticaemia in weanling pigs

2015 ◽  
Vol 82 (1) ◽  
Author(s):  
Folorunso O. Fasina ◽  
Dauda G. Bwala ◽  
Evelyn Madoroba
Keyword(s):  
Escherichia Coli ◽  
South African ◽  
Multidrug Resistant ◽  
Weanling Pigs ◽  
Tissue Samples ◽  
E Coli ◽  
Young Pigs ◽  
Polymerase Chain ◽  
The Impact ◽  
Future Management

Escherichia coli is usually a benign commensal of the gut microflora. However, when E. coli acquires virulence genes it can multiply rapidly and cause disease through colonisation of the intestinal mucosa. Escherichia coli can become a significant pathogen in young pigs. We report an investigation of fatal colisepticaemia in weanling pigs from emerging farms where piglets and weaners were diarrhoeic and the mortality rate ranged between 15% and 70% in each litter. Faecal and tissue samples were processed for histopathology, bacteriology and molecular biology (multiplex and monoplex polymerase chain reaction) and we recovered enteroaggregative multidrug-resistant E. coli producing EAST-1 enterotoxin. An association between poor housing conditions and the observed cases was established and future management programmes were recommended to reduce the impact of such pathogens. Enteroaggregative E. coli is becoming a major problem in the pig industry. It therefore becomes necessary to establish the full impact of E. coli on the South African pig industry and to determine the geographic extent of the problem.

Survival of Escherichia coliO157:H7 in Synthetic Gastric Fluid after Cold and Acid Habituation in Apple Juice or Trypticase Soy Broth Acidified with Hydrochloric Acid or Organic Acids

1998 ◽  
Vol 61 (8) ◽  
pp. 939-947 ◽  
Author(s):  
HEIDI E. ULJAS ◽  
STEVEN C. INGHAM
Keyword(s):  
Lactic Acid ◽  
Citric Acid ◽  
Organic Acids ◽  
Apple Juice ◽  
Acid Tolerance ◽  
Storage Conditions ◽  
Gastric Fluid ◽  
E Coli ◽  
Storage Media ◽  
Strain Storage

Extreme acid tolerance of Escherichia coli O157:H7 has raised doubts about the safety of acidic foods. This study examined whether prior storage in acidic and/or cold conditions enhanced survival of E. coli O157 :H7 in synthetic gastric fluid (SGF). Three E. coli O157:H7 strains were stored in trypticase soy broth (TSB; acidified with HCl, malic acid, citric acid, or lactic acid) or pH 3.5 and 6.5 (nonacidic control) apple juice at 4 and 21°C for ≤7 days and then were incubated in pH 2.5 SGF at 37°C for 4 h. Cells survived better in apple juice than in TSB containing organic acids, suggesting that juice constituents other than organic acids protect E. coli O157:H7. Refrigeration combined with low pH best protected cells in apple juice and acidified TSB, but, compared to the nonacidic control, only acidified TSB enhanced subsequent survival in pH 2.5 SGF. Equal survival in SGF occurred after storage in pH 3.5 or 6.5 apple juice at 4°C, suggesting that low temperature alone in apple juice enhanced acid tolerance. Two strains stored at 4°C in TSB containing malic or citric acid subsequently survived better in SGF than cells stored in nonacidified TSB but poorer than cells stored in the presence of HCl. These differences reflect the higher pKa of these organic acids. However, subsequent survival of these strains in SGF was poorer after refrigerated storage in apple juice than in TSB containing citric or malic acids. Cells stored in lactic acid were most likely to be completely eliminated upon transfer to SGF. Differences in survival in storage media or SGF related to strain, storage conditions, or acidifier were consistent and often statistically significant (P < 0.05). Although the survival of E. coli O157:H7 in refrigerated acidic beverages may not be affected by the type of acidifier used, the subsequent survival in SGF of this pathogen may be critically dependent on this factor.

scholarly journals Growth and Survival of Acid-Resistant and Non-Acid-Resistant Shiga-Toxin-ProducingEscherichia coliStrains during the Manufacture and Ripening of Camembert Cheese

2009 ◽  
Vol 2009 ◽  
pp. 1-10 ◽  
Author(s):  
M. P. Montet ◽  
E. Jamet ◽  
S. Ganet ◽  
M. Dizin ◽  
S. Miszczycha ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Acid Tolerance ◽  
Acid Resistance ◽  
Gastric Fluid ◽  
Simulated Gastric Fluid ◽  
Growth And Survival ◽  
Cheese Making ◽  
Early Stages ◽  
Camembert Cheese

Growth and survival of acid-resistant (AR) and non-acid-resistant (NAR) Shiga-toxin-producingEscherichia coli(STEC) strains were investigated during the manufacture and ripening of microfiltered milk Camembert cheeses. The induction of acid resistance of the STEC strains in cheeses was also studied. Six different mixtures of AR and/or NAR STEC strains were inoculated separately into microfiltered milk at a level of103CFUmL−1. The STEC counts (AR and NAR) initially increased by 1 to 2log⁡10CFUg−1during cheese-making. Thereafter, the populations stabilized during salting/drying and then decreased during the early stages of ripening. Exposing the STEC strains in artificially inoculated cheeses to simulated gastric fluid (SGF - pH: 2.0) reduced the number of NAR strains to undetectable levels within 40 minutes, versus 120 minutes for the AR STEC strains. AR and NAR STEC were able to survive during the manufacture and ripening of Camembert cheese prepared from microfiltered milk with no evidence of induced acid tolerance in NAR STEC strains.

scholarly journals Stress Resistance Development and Genome-Wide Transcriptional Response ofEscherichia coliO157:H7 Adapted to Sublethal Thymol, Carvacrol, andtrans-Cinnamaldehyde

2018 ◽  
Vol 84 (22) ◽  
Author(s):  
Wenqian Yuan ◽  
Zi Jing Seng ◽  
Gurjeet Singh Kohli ◽  
Liang Yang ◽  
Hyun-Gyun Yuk
Keyword(s):  
Escherichia Coli ◽  
Stress Resistance ◽  
Virulence Genes ◽  
Acid Resistance ◽  
Gastric Fluid ◽  
Simulated Gastric Fluid ◽  
Cross Resistance ◽  
Resistance Development ◽  
Content Type ◽  
E Coli

ABSTRACTThymol, carvacrol, andtrans-cinnamaldehyde are essential oil (EO) compounds with broad-spectrum antimicrobial activities against foodborne pathogens, includingEscherichia coliO157:H7. However, little is known regarding direct resistance and cross-resistance development inE. coliO157:H7 after adaptation to sublethal levels of these compounds, and information is scarce on microbial adaptive responses at a molecular level. The present study demonstrated thatE. coliO157:H7 was able to grow in the presence of sublethal thymol (1/2T), carvacrol (1/2C), ortrans-cinnamaldehyde (1/2TC), displaying an extended lag phase duration and a lower maximum growth rate. EO-adapted cells developed direct resistance against lethal EO treatments and cross-resistance against heat (58°C) and oxidative (50 mM H2O2) stresses. However, no induction of acid resistance (simulated gastric fluid, pH 1.5) was observed. RNA sequencing revealed a large number (310 to 338) of differentially expressed (adjustedPvalue [Padj], <0.05; fold change, ≥5) genes in 1/2T and 1/2C cells, while 1/2TC cells only showed 27 genes with altered expression. In accordance with resistance phenotypes, the genes related to membrane, heat, and oxidative stress responses and genes related to iron uptake and metabolism were upregulated. Conversely, virulence genes associated with motility, biofilm formation, and efflux pumps were repressed. This study demonstrated the development of direct resistance and cross-resistance and characterized whole-genome transcriptional responses inE. coliO157:H7 adapted to sublethal thymol, carvacrol, ortrans-cinnamaldehyde. The data suggested that caution should be exercised when using EO compounds as food antimicrobials, due to the potential stress resistance development inE. coliO157:H7.IMPORTANCEThe present study was designed to understand transcriptomic changes and the potential development of direct and cross-resistance in essential oil (EO)-adaptedEscherichia coliO157:H7. The results demonstrated altered growth behaviors ofE. coliO157:H7 during adaptation in sublethal thymol, carvacrol, andtrans-cinnamaldehyde. Generally, EO-adapted bacteria showed enhanced resistance against subsequent lethal EO, heat, and oxidative stresses, with no induction of acid resistance in simulated gastric fluid. A transcriptomic analysis revealed the upregulation of related stress resistance genes and a downregulation of various virulence genes in EO-adapted cells. This study provides new insights into microbial EO adaptation behaviors and highlights the risk of resistance development in adapted bacteria.

scholarly journals The Protein Interactome of Glycolysis in Escherichia coli

Proteomes ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 16
Author(s):  
Shomeek Chowdhury ◽  
Stephen Hepper ◽  
Mudassir K. Lodi ◽  
Milton H. Saier ◽  
Peter Uetz
Keyword(s):  
Escherichia Coli ◽  
Protein Interactions ◽  
Allosteric Regulation ◽  
Glycolytic Enzymes ◽  
Protein Protein Interactions ◽  
Post Translational Modification ◽  
Interacting Protein ◽  
E Coli ◽  
Protein Interactome ◽  
The Impact

Glycolysis is regulated by numerous mechanisms including allosteric regulation, post-translational modification or protein-protein interactions (PPI). While glycolytic enzymes have been found to interact with hundreds of proteins, the impact of only some of these PPIs on glycolysis is well understood. Here we investigate which of these interactions may affect glycolysis in E. coli and possibly across numerous other bacteria, based on the stoichiometry of interacting protein pairs (from proteomic studies) and their conservation across bacteria. We present a list of 339 protein-protein interactions involving glycolytic enzymes but predict that ~70% of glycolytic interactors are not present in adequate amounts to have a significant impact on glycolysis. Finally, we identify a conserved but uncharacterized subset of interactions that are likely to affect glycolysis and deserve further study.

Modelling inactivation of wild‐type and clinical Escherichia coli O26 strains using UV‐C and thermal treatment and subsequent persistence in simulated gastric fluid

2019 ◽  
Vol 127 (5) ◽  
pp. 1564-1575 ◽  
Author(s):  
V.S. Castro ◽  
D.K.A. Rosario ◽  
Y.S. Mutz ◽  
A.C.C. Paletta ◽  
E.E.S. Figueiredo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Thermal Treatment ◽  
Gastric Fluid ◽  
Simulated Gastric Fluid ◽  
Wild Type ◽  
Uv C

scholarly journals Solubility assessment of single-chain antibody fragment against epithelial cell adhesion molecule extracellular domain in four Escherichia coli strains

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Fatemeh Sadat Javadian ◽  
Majid Basafa ◽  
Aidin Behravan ◽  
Atieh Hashemi
Keyword(s):  
Escherichia Coli ◽  
Cell Adhesion ◽  
Epithelial Cell ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Extracellular Domain ◽  
Epithelial Cell Adhesion Molecule ◽  
Single Chain ◽  
E Coli ◽  
The Impact

Abstract Background Overexpression of the EpCAM (epithelial cell adhesion molecule) in malignancies makes it an attractive target for passive immunotherapy in a wide range of carcinomas. In comparison with full-length antibodies, due to the small size, the scFvs (single-chain variable fragments) are more suitable for recombinant expression in E. coli (Escherichia coli). However, the proteins expressed in large amounts in E. coli tend to form inclusion bodies that need to be refolded which may result in poor recovery of bioactive proteins. Various engineered strains were shown to be able to alleviate the insolubility problem. Here, we studied the impact of four E. coli strains on the soluble level of anti-EpEX-scFv (anti-EpCAM extracellular domain-scFv) protein. Results Although results showed that the amount of soluble anti-EpEX-scFv obtained in BL21TM (DE3) (114.22 ± 3.47 mg/L) was significantly higher to those produced in the same condition in E. coli RosettaTM (DE3) (71.39 ± 0.31 mg/L), and OrigamiTM T7 (58.99 ± 0.44 mg/L) strains, it was not significantly different from that produced by E. coli SHuffleTM T7 (108.87 ± 2.71 mg/L). Furthermore, the highest volumetric productivity of protein reached 318.29 ± 26.38 mg/L in BL21TM (DE3). Conclusions Although BL21TM (DE3) can be a suitable strain for high-level production of anti-EpEX-scFv protein, due to higher solubility yield (about 55%), E. coli SHuffleTM T7 seems to be better candidate for soluble production of scfv compared to BL21TM (DE3) (solubility yield of about 30%).

One-Step Preparation of Competent E. coli: Transformation and Storage of Bacterial Cells in the Same Solution

2021 ◽  
Vol 2021 (11) ◽  
pp. pdb.prot101212 ◽  
Author(s):  
Michael R. Green ◽  
Joseph Sambrook
Keyword(s):  
Escherichia Coli ◽  
Convenient Method ◽  
Plasmid Dna ◽  
Transformation Efficiency ◽  
Bacterial Cells ◽  
E Coli ◽  
One Step ◽  
And Storage

This protocol describes a convenient method for the preparation, use, and storage of competent Escherichia coli. The reported transformation efficiency of this method is ∼5 × 107 transformants/µg of plasmid DNA.

scholarly journals Impact of Vacuum Cooling on Escherichia coli O157:H7 Infiltration into Lettuce Tissue

2008 ◽  
Vol 74 (10) ◽  
pp. 3138-3142 ◽  
Author(s):  
Haiping Li ◽  
Mehrdad Tajkarimi ◽  
Bennie I. Osburn
Keyword(s):  
Escherichia Coli ◽  
Risk Assessment ◽  
Reduction Rate ◽  
Escherichia Coli O157 ◽  
Surface Sterilization ◽  
E Coli ◽  
Green Industry ◽  
Vacuum Cooling ◽  
The Impact

ABSTRACT Vacuum cooling is a common practice in the California leafy green industry. This study addressed the impact of vacuum cooling on the infiltration of Escherichia coli O157:H7 into lettuce as part of the risk assessment responding to the E. coli O157:H7 outbreaks associated with leafy green produce from California. Vacuum cooling significantly increased the infiltration of E. coli O157:H7 into the lettuce tissue (2.65E+06 CFU/g) compared to the nonvacuumed condition (1.98E+05 CFU/g). A stringent surface sterilization and quadruple washing could not eliminate the internalized bacteria from lettuce. It appeared that vacuuming forcibly changed the structure of lettuce tissue such as the stomata, suggesting a possible mechanism of E. coli O157:H7 internalization. Vacuuming also caused a lower reduction rate of E. coli O157:H7 in stored lettuce leaves than that for the nonvacuumed condition.

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.

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