scholarly journals Are Antimicrobial Interventions Associated with Heat-Resistant Escherichia coli on Meat?

2020 ◽  
Vol 86 (13) ◽  
Author(s):  
Peipei Zhang ◽  
Frances Tran ◽  
Kim Stanford ◽  
Xianqin Yang

ABSTRACT Decontamination practices, which often involve thermal treatments, are routinely performed in beef packing plants and have generally improved the safety of meat in North America. We investigated whether Escherichia coli in the beef production chain is becoming more heat resistant due to those treatments. Cattle isolates (n = 750) included seven serogroups (O157, O103, O111, O121, O145, O26, and O45) which were collected between 2002 and 2017. Beef plant isolates (n = 700) from carcasses, fabrication equipment, and beef products were included. Heat resistance was determined in Luria-Bertani broth at 60°C and by PCR screening for the locus of heat resistance (LHR). The decimal reduction for E. coli at 60°C (D60ºC values) ranged from 0 to 7.54 min, with 97.2% of the values being <2 min. The prevalence of E. coli with D60ºC values of >2 min was not significantly different (P > 0.05) among cattle and meat plant isolates. E. coli from equipment before sanitation (median, 1.03 min) was more heat resistant than that after sanitation (median, 0.9 min). No significant difference in D60ºC values was observed among E. coli isolates from different years, from carcasses before and after antimicrobial interventions, or from before and during carcass chilling. Of all isolates, 1.97% harbored LHR, and the LHR-positive isolates had greater median D60ºC values than the LHR-negative isolates (3.25 versus 0.96 min). No increase in heat resistance in E. coli was observed along the beef production chain or with time. IMPORTANCE The implementation of multiple hurdles in the beef production chain has resulted in substantial improvement in the microbial safety of beef in Canada. In this study, we characterized a large number of Escherichia coli isolates (n = 1,450) from various sources/stages of beef processing to determine whether the commonly used antimicrobial interventions would give rise to heat-resistant E. coli on meat, which in turn may require alternatives to the current control of pathogens and/or modifications to the current cooking recommendations for meat. The findings show that the degree and rate of heat resistance in E. coli did not increase along the production chain or with time. This furthers our understanding of man-made ecological niches that are required for the development of heat resistance in E. coli.

2017 ◽  
Vol 83 (20) ◽  
Author(s):  
Ryan Mercer ◽  
Oanh Nguyen ◽  
Qixing Ou ◽  
Lynn McMullen ◽  
Michael G. Gänzle

ABSTRACT The locus of heat resistance (LHR) is a 15- to 19-kb genomic island conferring exceptional heat resistance to organisms in the family Enterobacteriaceae, including pathogenic strains of Salmonella enterica and Escherichia coli. The complement of LHR-comprising genes that is necessary for heat resistance and the stress-induced or growth-phase-induced expression of LHR-comprising genes are unknown. This study determined the contribution of the seven LHR-comprising genes yfdX1 GI, yfdX2, hdeD GI, orf11, trx GI, kefB, and psiE GI by comparing the heat resistances of E. coli strains harboring plasmid-encoded derivatives of the different LHRs in these genes. (Genes carry a subscript “GI” [genomic island] if an ortholog of the same gene is present in genomes of E. coli.) LHR-encoded heat shock proteins sHSP20, ClpKGI, and sHSPGI are not sufficient for the heat resistance phenotype; YfdX1, YfdX2, and HdeD are necessary to complement the LHR heat shock proteins and to impart a high level of resistance. Deletion of trx GI, kefB, and psiE GI from plasmid-encoded copies of the LHR did not significantly affect heat resistance. The effect of the growth phase and the NaCl concentration on expression from the putative LHR promoter p2 was determined by quantitative reverse transcription-PCR and by a plasmid-encoded p2:GFP promoter fusion. The expression levels of exponential- and stationary-phase E. coli cells were not significantly different, but the addition of 1% NaCl significantly increased LHR expression. Remarkably, LHR expression in E. coli was dependent on a chromosomal copy of evgA. In conclusion, this study improved our understanding of the genes required for exceptional heat resistance in E. coli and factors that increase their expression in food. IMPORTANCE The locus of heat resistance (LHR) is a genomic island conferring exceptional heat resistance to several foodborne pathogens. The exceptional level of heat resistance provided by the LHR questions the control of pathogens by current food processing and preparation techniques. The function of LHR-comprising genes and their regulation, however, remain largely unknown. This study defines a core complement of LHR-encoded proteins that are necessary for heat resistance and demonstrates that regulation of the LHR in E. coli requires a chromosomal copy of the gene encoding EvgA. This study provides insight into the function of a transmissible genomic island that allows otherwise heat-sensitive enteric bacteria, including pathogens, to lead a thermoduric lifestyle and thus contributes to the detection and control of heat-resistant enteric bacteria in food.


1980 ◽  
Vol 43 (5) ◽  
pp. 376-380 ◽  
Author(s):  
R. S. SINGH ◽  
B. RANGANATHAN

Three Escherichia coli cultures (0111:B4, 0127:B8 and NP) were selected to study their heat-resistant characteristics when in cow skim, cow whole and buffalo whole milk. The temperatures of heat-treatment included in this study were 50, 55, 60 and 63 C. The time interval during heat-treatment was 10 min at 50 and 55 C and 5 min at 60 and 63 C. Marked differences in heat-resistance were observed in the three E. coli cultures. The z-values obtained for strain 0111:B4 were 8.3, 9.0 and 10.2 when tested in cow skim milk, cow whole milk and buffalo milk, respectively. The z-values for 0127:B8 and NP were 17.5, 18.0 and 19.2 and 18.8, 19.0 and 20.3, respectively, for the three types of milk.


2020 ◽  
Vol 86 (14) ◽  
Author(s):  
Subhrajit Bhar ◽  
Tungadri Bose ◽  
Sharmila S. Mande

ABSTRACT Signal transduction systems are essential for microorganisms to respond to their ever-changing environment. They can be distinguished into one-component systems, two-component systems, and extracytoplasmic-function σ factors. Abundances of a few signal-transducing proteins, termed herein as sensory proteins (SPs), have previously been reported to be correlated with the genome size and ecological niche of certain Gram-positive bacteria. No such reports are available for Gram-negative bacteria. The current study attempts to investigate the relationship of the abundances of SPs to genome size in Escherichia coli, and the bacterial pathotypes or phylotypes. While the relationship between SP abundance and genome size could not be established, the sensory protein index (SPI), a new metric defined herein, was found to be correlated with E. coli virulence. In addition, significant association was observed among the distribution of SPs and E. coli pathotypes. Results indicate that such associations might be due to genomic rearrangements to best utilize the resources available in a given ecological niche. Overall, the study provides an in-depth analysis of the occurrence of different SPs among pathogenic and nonpathogenic E. coli strains. Possibilities of using the SPI as a marker for identifying pathogenic strains from among an organism complex are also discussed. IMPORTANCE Sensory proteins (SPs) act as sensors and actuators for a cell and participate in important mechanisms pertaining to bacterial survival, adaptation, and virulence. Therefore, bacterial species residing in similar ecological niches or those sharing common pathotypes are expected to exhibit similar SP signatures. We have investigated profiles of SPs in different species of Escherichia coli and present in this article the sensory protein index (SPI), a metric for quantifying the abundance and/or distribution of SPs across bacterial genomes, which could indicate the virulence potency of a bacterium. The SPI could find use in characterizing uncultured strains and bacterial complexes, as a biomarker for disease diagnostics, evaluating the effect of therapeutic interventions, assessing effects of ecological alterations, etc. Grouping the studied strains of E. coli on the basis of the frequency of occurrence of SPs in their genomes could potentially replicate the stratification of these strains on the basis of their phylotypes. In addition, E. coli strains belonging to the same pathotypes were also seen to share similar SP signatures. Furthermore, the SPI was seen to be an indicator of pathogenic potency of E. coli strains. The SPI metric is expected to be useful in the (pathogenic) characterization of hereto uncultured strains which are routinely sequenced in host microbiome analysis projects, or from among an ensemble of microbial organisms constituting a biospecimen. Thus, the possibilities of using the SPI as a biomarker for diagnosis of a disease or the outcome of a therapeutic intervention cannot be ruled out. Further, SPIs obtained from longitudinal ecological samples have the potential to serve as key indicators of environmental changes. Such changes in the environment are often detrimental to the resident biome and methods for timely detection of environmental changes hold huge socioeconomic benefits.


2014 ◽  
Vol 80 (14) ◽  
pp. 4383-4390 ◽  
Author(s):  
Ron N. Xavier ◽  
Hugh W. Morgan ◽  
Ian R. McDonald ◽  
Helen Withers

ABSTRACTThe ability to maintain a dual lifestyle of colonizing the ruminant gut and surviving in nonhost environments once shed is key to the success ofEscherichia coliO157:H7 as a zoonotic pathogen. Both physical and biological conditions encountered by the bacteria are likely to change during the transition between host and nonhost environments. In this study, carbon starvation at suboptimal temperatures in nonhost environments was simulated by starving a New Zealand bovineE. coliO157:H7 isolate in phosphate-buffered saline at 4 and 15°C for 84 days. Recovery of starved cells on media with different nutrient availabilities was monitored under aerobic and anaerobic conditions. We found that the New Zealand bovineE. coliO157:H7 isolate was able to maintain membrane integrity and viability over 84 days and that the level of recovery depended on the nutrient level of the recovery medium as well as the starvation temperature. In addition, a significant difference in carbon utilization was observed between starved and nonstarved cells.


2011 ◽  
Vol 55 (6) ◽  
pp. 3005-3007 ◽  
Author(s):  
Ivan Literak ◽  
Radim Petro ◽  
Monika Dolejska ◽  
Erika Gruberova ◽  
Hana Dobiasova ◽  
...  

ABSTRACTThe study was performed in the Czech Republic during 2007 to 2009. OfEscherichia coliisolates from 275 children aged 6 weeks, 36% (n= 177) were resistant to 1 to 7 antibiotics. Of isolates from 253 children aged 6 to 17 years, 24% (n= 205) were resistant to 1 to 5 antibiotics. There was no significant difference in the prevalences of antibiotic-resistantE. coliisolates between these groups of children, even though the consumptions of antibiotics were quite different.


1998 ◽  
Vol 61 (9) ◽  
pp. 1184-1186 ◽  
Author(s):  
NICOLE C. WILLIAMS ◽  
STEVEN C. INGHAM

On a beef carcass, Escherichia coli may sequentially encounter acid- and heat-intervention steps. This study tested whether acid stress (1.5% [vol/vol] acetic acid, pH 4.0, 37°C, 15 min) would enhance subsequent heat resistance of E. coli. Initially, cells (E. coli O157:H7 ATCC 43894, nonpathogenic E. coli B [strain FRIK-124], and rpoS-deficient mutant 813-6 [derived from E. coli O157:H7 ATCC 43895]) were acid stressed and transferred to 54°C tiypticase soy broth (TSB), and survivors were immediately enumerated after at least three intervals of 12, 2, and 6 min, respectively, by plating. The ATCC 43894 and 813-6 strains survived the acid stress but strain FRIK-124 did not. Acid-stressed ATCC 43894 had significantly lower D values than the non-acid-stressed controls. Strain 813-6 had significantly lower D values than strain ATCC 43894, with no significant difference between acid-stressed and non-acid-stressed cells. In a second experiment, cooling of cells prior to plating resulted in an increased D value for acid-stressed ATCC 43894 cells, such that it was not significantly different from the D value for non-acid-stressed Controls. Using this protocol, there was no significant difference in D values between acid-stressed and non-acid-stressed ATCC 43894 cells in prewarmed TSB (54, 58, and 62°C), in prewarmed ground beef slurry (GBS; 58°C), or in TSB and GBS inoculated at 5°C and heated to 58°C. The acid stress tested does not enhance subsequent heat resistance of E. coli.


2011 ◽  
Vol 77 (20) ◽  
pp. 7339-7344 ◽  
Author(s):  
R. M. Goulter-Thorsen ◽  
E. Taran ◽  
I. R. Gentle ◽  
K. S. Gobius ◽  
G. A. Dykes

ABSTRACTThe role of curli expression in attachment ofEscherichia coliO157:H7 to glass, Teflon, and stainless steel (SS) was investigated through the creation ofcsgAknockout mutants in two isolates ofE. coliO157:H7. Attachment assays using epifluorescence microscopy and measurements of the force of adhesion of bacterial cells to the substrates using atomic force microscopy (AFM) force mapping were used to determine differences in attachment between wild-type (wt) andcsgA-negative (ΔcsgA) strains following growth in four different media. The hydrophobicity of the cells was determined using contact angle measurements (CAM) and bacterial adhesion to hydrocarbons (BATH). The attachment assay results indicated that ΔcsgAstrains attached to glass, Teflon, and SS surfaces in significantly different numbers than their wt counterparts in a growth medium-dependent fashion (P< 0.05). However, no clear correlation was seen between attachment numbers, surface type, or growth medium. No correlation was seen between BATH and CAM results (R2< 0.70). Hydrophobicity differed between the wt and ΔcsgAin some cases in a growth medium- and method-dependent fashion (P< 0.05). AFM force mapping revealed no significant difference in the forces of adhesion to glass and SS surfaces between wt and ΔcsgAstrains (P> 0.05) but a significantly greater force of adhesion to Teflon for one of the two wt strains than for its ΔcsgAcounterpart (P< 0.05). This study shows that CsgA production byE. coliO157:H7 may alter attachment behavior in some environments; however, further investigation is required in order to determine the exact relationship between CsgA production and attachment to abiotic surfaces.


2019 ◽  
Vol 3 (2) ◽  
Author(s):  
X. Yang ◽  
F. Tran ◽  
M. Klasse

ObjectivesRecent reports of an extremely heat resistant but non-pathogenic beef Escherichia coli strain, AW 1.7, raised concerns over the adequacy of cooking ground beef to 71°C in Canada. The objective of this study was to assess the adequacy of the current cooking recommendations for ground beef in relation to heat resistant E. coli.Materials and MethodsIn total, 8 potentially heat resistant E. coli strains (4 generic and 4 E. coli O157:H7) from beef along with E. coli AW1.7 were included in this study. Heat resistance of the strains was first evaluated by decimal reductions at 60°C (D60°C-value), the time required to have a log reduction of the bacterial population at 60°C. The more heat resistant strains of each group (E. coli 62 and 68, and E. coli O157 J3 and C37) were further assessed for their heat resistance when grown in Lennox Broth without salt (LB-NS), LB + 2% NaCl and Meat Juice (MJ). Then, the two most heat resistant E. coli O157 strains (J3 and C37) and E. coli AW 1.7 were each introduced to extra lean ground beef (100 g) in vacuum pouches for determination of their D-values at three temperatures, 54, 57, and 60°C, from which a z-value for each strain was derived. The thermal characteristics of all three strains were fed into a predictive model to determine the process lethality of cooking burgers to 71°C with resting for up to 5 min. Finally, inactivation of the most heat resistant E. coli strain AW1.7, assessed in this study and reported in the literature, in ground beef was validated by grilling burgers containing 6.20 ± 0.24 log CFU/g of the organism to 71°C without or with a resting of 3 or 5 min.ResultsThe D60°C-values for these strains varied from 1.3 to 9.0 min, with J3 and AW1.7 being the least and most heat resistant strains, respectively. The D60°C-values for E. coli 62 and 68 were similar and were not affected by growth medium, while the heat resistance of C37, J3 and AW1.7 varied with the growth medium. When heated in extra lean ground beef (100 g) in vacuum pouches, the mean D54°C, D57°C, and D60°C-values were 44.8, 18.6 and 2.9 min for C37, 13.8, 6.9 and 0.9 min for J3, and 40.5, 9.1 6.1 min for AW1.7. The derived z- and D71°C-values were, respectively, 5.0, 5.1 and 7.3°C; and 0.022, 0.008, and 0.156 min. Burger temperatures continued to rise after being removed from heat when the target temperature was reached, by up to 5°C, and resting of 1 min would result in a destruction of 133, 374, and 14 log C37, J3 and AW1.7, estimated from process lethality. When burgers inoculated with AW1.7 were cooked to 71°C, 14 of the 15 burgers yielded no E. coli, while the 15th had a reduction of 4.5 log. Additional resting of 3 or 5 min resulted in complete elimination of AW 1.7.ConclusionIt has been predicted that 2% of E. coli from beef may carry heat resistant genes. The findings in this study, along with the very low level of total E. coli expected in ground beef in Canada, suggest that cooking ground beef to 71°C should be adequate to ensure the safety of such products.


2014 ◽  
Vol 80 (23) ◽  
pp. 7230-7234 ◽  
Author(s):  
Charlotte Valat ◽  
Karine Forest ◽  
Frédéric Auvray ◽  
Véronique Métayer ◽  
Thomas Méheut ◽  
...  

ABSTRACTThe CS31A, F17, and F5 adhesins are usually targeted by serology-based methods to detect pathogenicEscherichia coliassociated with calf enteritis. However, the virulence traits of the selected isolates are still poorly described. Here, from a set of 349 diarrheagenicE. coliisolates from cattle, we demonstrated a 70.8% concordance rate (Cohen's kappa, 0.599) between serology- and PCR-based approaches for the detection of adhesins under field conditions. A 79% to 82.4% correspondence between the two methods was found for fimbrial adhesins, whereas major discrepancies (33%) were observed for CS31A-type antigens. Various F17A variants were found, such as F17Ac (20K) (50%), F17Aa (FY) (18.9%), F17Ab (8.1%), and F17Ad (111K) (5.4%), including a high proportion (17.6%) of new F17A internal combinations (F17Aab, F17Aac, and F17Abc) or untypeable variants. In addition, the highest proportion of pathovar-associated virulence factor (VF) genes was observed amongE. coliisolates that produced F5/F41 adhesins. A specific link between the heat-stable toxins related to the enterotoxigenicE. coli(ETEC) pathovar and adhesins was identified. STa was significantly linked to F5/F41 and EAST1 to CS31A adhesins (P< 0.001), respectively, whereas NTEC was associated with F17 adhesin (P= 0.001). Clustering between phylogroups according to the adhesin types was also observed. Also, few Shiga toxin-producingE. coli(STEC) or enteropathogenicE. coli(EPEC) pathovars were identified. Finally, no statistically significant difference was observed in the occurrence of extended-spectrum beta lactamase (ESBL) production according to the adhesins expressed by the isolates (P= 0.09). Altogether, this study gives new insights into the relationship between adhesins, VF, and antimicrobial resistance in calf enteritis and supports the need for further standardization of methodologies for such approaches.


2013 ◽  
Vol 57 (5) ◽  
pp. 2109-2113 ◽  
Author(s):  
F. Docobo-Pérez ◽  
L. López-Cerero ◽  
R. López-Rojas ◽  
P. Egea ◽  
J. Domínguez-Herrera ◽  
...  

ABSTRACTEscherichia coliis commonly involved in infections with a heavy bacterial burden. Piperacillin-tazobactam and carbapenems are among the recommended empirical treatments for health care-associated complicated intra-abdominal infections. In contrast to amoxicillin-clavulanate, both have reducedin vitroactivity in the presence of high concentrations of extended-spectrum β-lactamase (ESBL)-producing and non-ESBL-producingE. colibacteria. Our goal was to compare the efficacy of these antimicrobials against different concentrations of two clinicalE. colistrains, one an ESBL-producer and the other a non-ESBL-producer, in a murine sepsis model. An experimental sepsis model {∼5.5 log10CFU/g [low inoculum concentration (LI)] or ∼7.5 log10CFU/g [high inoculum concentration (HI)]} usingE. colistrains ATCC 25922 (non-ESBL producer) and Ec1062 (CTX-M-14 producer), which are susceptible to the three antimicrobials, was used. Amoxicillin-clavulanate (50/12.5 mg/kg given intramuscularly [i.m.]), piperacillin-tazobactam (25/3.125 mg/kg given intraperitoneally [i.p.]), and imipenem (30 mg/kg i.m.) were used. Piperacillin-tazobactam and imipenem reduced spleen ATCC 25922 strain concentrations (−2.53 and −2.14 log10CFU/g [P< 0.05, respectively]) in the HI versus LI groups, while amoxicillin-clavulanate maintained its efficacy (−1.01 log10CFU/g [no statistically significant difference]). Regarding the Ec1062 strain, the antimicrobials showed lower efficacy in the HI than in the LI groups: −0.73, −1.89, and −1.62 log10CFU/g (P< 0.05, for piperacillin-tazobactam, imipenem, and amoxicillin-clavulanate, respectively, although imipenem and amoxicillin-clavulanate were more efficacious than piperacillin-tazobactam). An adapted imipenem treatment (based on the time for which the serum drug concentration remained above the MIC obtained with a HI of the ATCC 25922 strain) improved its efficacy to −1.67 log10CFU/g (P< 0.05). These results suggest that amoxicillin-clavulanate could be an alternative to imipenem treatment of infections caused by ESBL- and non-ESBL-producingE. colistrains in patients with therapeutic failure with piperacillin-tazobactam.


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