scholarly journals Inactivation of Shiga Toxin-Producing Escherichia coli (STEC) and Degradation and Removal of Cellulose from STEC Surfaces by Using Selected Enzymatic and Chemical Treatments

2011 ◽  
Vol 77 (24) ◽  
pp. 8532-8537 ◽  
Author(s):  
Yoen Ju Park ◽  
Jinru Chen

ABSTRACTSome Shiga toxin-producingEscherichia coli(STEC) strains produce extracellular cellulose, a long polymer of glucose with β-1-4 glycosidic bonds. This study evaluated the efficacies of selected enzymatic and chemical treatments in inactivating STEC and degrading/removing the cellulose on STEC surfaces. Six cellulose-producing STEC strains were treated with cellulase (0.51 to 3.83 U/15 ml), acetic and lactic acids (2 and 4%), as well as an acidic and alkaline sanitizer (manufacturers' recommended concentrations) under appropriate conditions. Following each treatment, residual amounts of cellulose and surviving populations of STEC were determined. Treatments with acetic and lactic acids significantly (P< 0.05) reduced the populations of STEC, and those with lactic acid also significantly decreased the amounts of cellulose on STEC. The residual amounts of cellulose on STEC positively correlated to the surviving populations of STEC after the treatments with the organic acids (r= 0.64 to 0.94), and the significance of the correlations ranged from 83 to 99%. Treatments with cellulase and the sanitizers both degraded cellulose. However, treatments with cellulase had no influence on the fate of STEC, and those with the sanitizers reduced STEC cell populations to undetectable levels. Thus, the correlations between the residual amounts of cellulose and the surviving populations of STEC caused by these two treatments were not observed. The results suggest that the selected enzymatic and chemical agents degraded and removed the cellulose on STEC surfaces, and the treatments with organic acids and sanitizers also inactivated STEC cells. The amounts of cellulose produced by STEC strains appear to affect their susceptibilities to certain sanitizing treatments.

2011 ◽  
Vol 74 (5) ◽  
pp. 820-825 ◽  
Author(s):  
YOEN JU PARK ◽  
JINRU CHEN

This study was conducted to evaluate the abilities of five neutralizing agents, Dey-Engley (DE) neutralizing broth (single or double strength), morpholinepropanesulfonic acid (MOPS) buffer, phosphate-buffered saline (PBS), and sodium thiosulfate buffer, in mitigating the activities of acetic or lactic acid (2%) and an alkaline or acidic sanitizer (a manufacturer-recommended concentration) againt the cells of Shiga toxin–producing Escherichia coli (STEC; n = 9). To evaluate the possible toxicity of the neutralizing agents to the STEC cells, each STEC strain was exposed to each of the neutralizing agents at room temperature for 10 min. Neutralizing efficacy was evaluated by placing each STEC strain in a mixture of sanitizer and neutralizer under the same conditions. The neutralizing agents had no detectable toxic effect on the STEC strains. PBS was least effective for neutralizing the activity of selected organic acids and sanitizers. Single-strength DE and sodium thiosulfate neutralized the activity of both acetic and lactic acids. MOPS buffer neutralized the activity of acetic acid and lactic acid against six and five STEC strains, respectively. All neutralizing agents, except double-strength DE broth, had a limited neutralizing effect on the activity of the commercial sanitizers used in the study. The double-strength DE broth effectively neutralized the activity of the two commercial sanitizers with no detectable toxic effects on STEC cells.


2018 ◽  
Vol 81 (5) ◽  
pp. 762-768
Author(s):  
JOSHUA D. HASTY ◽  
JOHN A. HENSON ◽  
GARY R. ACUFF ◽  
DENNIS E. BURSON ◽  
JOHN B. LUCHANSKY ◽  
...  

ABSTRACT Scalding of hide-on bob veal carcasses with or without standard scalding chemical agents typically used for hogs, followed by an 82.2°C hot water wash and lactic acid spray (applied at ambient temperature) before chilling, was evaluated to determine its effectiveness in reducing Shiga toxin–producing Escherichia coli surrogate populations. A five-strain cocktail of rifampin-resistant, nonpathogenic E. coli surrogates was used to inoculate hides of veal carcasses immediately after exsanguination (target inoculation level of 7.0 log CFU/100 cm2). For carcasses receiving no scalding treatments, spraying with 82.2°C water as a final wash resulted in a 4.5-log CFU/100 cm2 surrogate reduction, and an additional 1.2-log CFU/100 cm2 reduction was achieved by spraying with 4.5% lactic acid before chilling. Scalding hide-on carcasses in 60°C water (no chemicals added) for 4 min in a traditional hog scalding tank resulted in a 2.1-log CFU/100 cm2 reduction in surrogate levels, and a subsequent preevisceration 82.2°C water wash provided an additional 2.9-log CFU/100 cm2 reduction. Spraying a 4.5% solution of lactic acid onto scalded, hide-on carcasses (after the 82.2°C water wash) resulted in a minimal additional reduction of 0.4 log CFU/100 cm2. Incorporation of scalding chemicals into the scald water resulted in a 4.1-log CFU/100 cm2 reduction (1.9 log CFU/100 cm2 greater than scalding without chemicals) in the surrogate population, and the first 82.2°C wash provided an additional 2.5-log CFU/100 cm2 reduction. Application of antimicrobial interventions did not affect the carcass temperature decline during chilling, the pH decline, or the color characteristics of the ribeye or the flank of the bob veal carcasses.


1991 ◽  
Vol 54 (7) ◽  
pp. 496-501 ◽  
Author(s):  
ARTHUR HINTON ◽  
GEORGE E. SPATES ◽  
DONALD E. CORRIER ◽  
MICHAEL E. HUME ◽  
JOHN R. DELOACH ◽  
...  

A Veillonella species and Enterococcus durans were isolated from the cecal contents of adult broilers. Mixed cultures of Veillonella and E. durans inhibited the growth of Salmonella typhimurium and Escherichia coli 0157:H7 on media containing 2.5% lactose (w/v). The growth of S. typhimurium or E. coli 0157:H7 was not inhibited by mixed cultures containing Veillonella and E. durans on media containing only 0.25% lactose or by pure cultures of Veillonella or E. durans on media containing either 0.25% or 2.5% lactose. The mixed cultures of Veillonella and E. durans produced significantly (P&lt;0.05) more acetic, propionic, and lactic acids in media containing 2.5% lactose than in media containing 0.25% lactose. The inhibition of the enteropathogens was related to the production of lactic acid from lactose by the E. durans and the production of acetic and propionic acids from lactic acid by the Veillonella.


1990 ◽  
Vol 53 (8) ◽  
pp. 652-655 ◽  
Author(s):  
DONALD E. CONNER ◽  
VIRGINIA N. SCOTT ◽  
DANE T. BERNARD

Growth and survival of four strains of Listeria monocytogenes under acidic conditions were investigated. Tryptic soy broth with yeast extract (TSBYE) was acidified with acetic, citric, hydrochloric, lactic, or propionic acid to pH 4.0–6.0, inoculated with L. monocytogenes and incubated at 30 or 4°C. The minimum test pH at which L. monocytogenes did not grow (inhibitory pH) was determined for each acid. In the pH range tested, this inhibitory pH was 5.0 for propionic acid, 4.5 for acetic and lactic acids, and 4.0 for citric and hydrochloric acids. All four strains gave similar results. Subsequent studies were conducted at 10 and 30°C to determine changes in cell populations in TSBYE adjusted to each inhibitory pH. Initial populations of viable cells (104 CFU/ml) were reduced to &lt;10 CFU/ml within 1–3 weeks at 30°C, whereas at 10°C, L. monocytogenes survived for 11–12 weeks in acetic, citric, or propionic acid-adjusted media and for 6 weeks in media adjusted with hydrochloric or lactic acid. The concentration of undissociated lactic acid was 0.002 M at pH 4.5.


2015 ◽  
Vol 83 (6) ◽  
pp. 2338-2349 ◽  
Author(s):  
J. Funk ◽  
N. Biber ◽  
M. Schneider ◽  
E. Hauser ◽  
S. Enzenmüller ◽  
...  

In this study, the cytotoxicity of the recently described subtilase variant SubAB2-2of Shiga toxin-producingEscherichia coliwas determined and compared to the plasmid-encoded SubAB1and the chromosome-encoded SubAB2-1variant. The genes for the respective enzymatic active (A) subunits and binding (B) subunits of the subtilase toxins were amplified and cloned. The recombinant toxin subunits were expressed and purified. Their cytotoxicity on Vero cells was measured for the single A and B subunits, as well as for mixtures of both, to analyze whether hybrids with toxic activity can be identified. The results demonstrated that all three SubAB variants are toxic for Vero cells. However, the values for the 50% cytotoxic dose (CD50) differ for the individual variants. Highest cytotoxicity was shown for SubAB1. Moreover, hybrids of subunits from different subtilase toxins can be obtained which cause substantial cytotoxicity to Vero cells after mixing the A and B subunits prior to application to the cells, which is characteristic for binary toxins. Furthermore, higher concentrations of the enzymatic subunit SubA1exhibited cytotoxic effects in the absence of the respective B1subunit. A more detailed investigation in the human HeLa cell line revealed that SubA1alone induced apoptosis, while the B1subunit alone did not induce cell death.


2021 ◽  
Author(s):  
Timothy J. Dallman ◽  
David R. Greig ◽  
Saheer E. Gharbia ◽  
Claire Jenkins

Sequence similarity of pathogen genomes can infer the relatedness between isolates as the fewer genetic differences identified between pairs of isolates, the less time since divergence from a common ancestor. Clustering based on hierarchical single linkage clustering of pairwise SNP distances has been employed to detect and investigate outbreaks. Here, we evaluated the evidence-base for the interpretation of phylogenetic clusters of Shiga toxin-producing Escherichia coli (STEC) O157:H7. Whole genome sequences of 1193 isolates of STEC O157:H7 submitted to Public Health England between July 2015 and December 2016 were mapped to the Sakai reference strain. Hierarchical single linkage clustering was performed on the pairwise SNP difference between all isolates at descending distance thresholds. Cases with known epidemiological links fell within 5-SNP single linkage clusters. Five-SNP single linkage community clusters where an epidemiological link was not identified were more likely to be temporally and/or geographically related than sporadic cases. Ten-SNP single linkage clusters occurred infrequently and were challenging to investigate as cases were few, and temporally and/or geographically dispersed. A single linkage cluster threshold of 5-SNPs has utility for the detection of outbreaks linked to both persistent and point sources. Deeper phylogenetic analysis revealed that the distinction between domestic UK and imported isolates could be inferred at the sub-lineage level. Cases associated with domestically acquired infection that fall within clusters that are predominantly travel associated are likely to be caused by contaminated imported food.


2016 ◽  
Vol 79 (4) ◽  
pp. 561-568 ◽  
Author(s):  
NORASAK KALCHAYANAND ◽  
MOHAMMAD KOOHMARAIE ◽  
TOMMY L. WHEELER

ABSTRACT Several antimicrobial compounds are in commercial meat processing plants for pathogen control on beef carcasses. However, the efficacy of the method used is influenced by a number of factors, such as spray pressure, temperature, type of chemical and concentration, exposure time, method of application, equipment design, and the stage in the process that the method is applied. The objective of this study was to evaluate effectiveness of time of exposure of various antimicrobial compounds against nine strains of Shiga toxin–producing Escherichia coli (STEC) and four strains of Salmonella in aqueous antimicrobial solutions with and without organic matter. Non-O157 STEC, STEC O157:H7, and Salmonella were exposed to the following aqueous antimicrobial solutions with or without beef purge for 15, 30, 60, 120, 300, 600, and 1,800 s: (i) 2.5% lactic acid, (ii) 4.0% lactic acid, (iii) 2.5% Beefxide, (iv) 1% Aftec 3000, (v) 200 ppm of peracetic acid, (vi) 300 ppm of hypobromous acid, and (vii) water as a control. In general, increasing exposure time to antimicrobial compounds significantly (P ≤ 0.05) increased the effectiveness against pathogens tested. In aqueous antimicrobial solutions without organic matter, both peracetic acid and hypobromous acid were the most effective in inactivating populations of STEC and Salmonella, providing at least 5.0-log reductions with exposure for 15 s. However, in antimicrobials containing organic matter, 4.0% lactic acid was the most effective compound in reducing levels of STEC and Salmonella, providing 2- to 3-log reductions with exposure for 15 s. The results of this study indicated that organic matter and exposure time influenced the efficacy of antimicrobial compounds against pathogens, especially with oxidizer compounds. These factors should be considered when choosing an antimicrobial compound for an intervention.


2017 ◽  
Vol 61 (9) ◽  
Author(s):  
Andreas Bauwens ◽  
Lisa Kunsmann ◽  
Helge Karch ◽  
Alexander Mellmann ◽  
Martina Bielaszewska

ABSTRACT Ciprofloxacin, meropenem, fosfomycin, and polymyxin B strongly increase production of outer membrane vesicles (OMVs) in Escherichia coli O104:H4 and O157:H7. Ciprofloxacin also upregulates OMV-associated Shiga toxin 2a, the major virulence factor of these pathogens, whereas the other antibiotics increase OMV production without the toxin. These two effects might worsen the clinical outcome of infections caused by Shiga toxin-producing E. coli. Our data support the existing recommendations to avoid antibiotics for treatment of these infections.


2020 ◽  
Vol 9 (45) ◽  
Author(s):  
Yujie Zhang ◽  
Yen-Te Liao ◽  
Vivian C. H. Wu

ABSTRACT Shiga toxin-producing Escherichia coli (STEC) serotype O103 is one of the primary pathogenic contaminants of beef products, contributing to several foodborne outbreaks in recent years. Here, we report the whole-genome sequence of a STEC O103:H2 strain isolated from cattle feces that contains a locus of enterocyte effacement (LEE) pathogenicity island.


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