scholarly journals A Risk Assessment of Shiga Toxin-producing Escherichia coli (STEC) in the Norwegian Meat Chain with Emphasis on Dry-cured Sausages

2021 ◽  
pp. 52-54
Author(s):  
Espen Rimstad ◽  
Leiv Sigve Håvarstein ◽  
Georg Kapperud ◽  
Jørgen Lassen ◽  
Bjørn-Tore Lunestad ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Risk Assessment ◽  
Food Safety ◽  
Shiga Toxin ◽  
Starter Cultures ◽  
Epidemiological Surveillance ◽  
Contamination Level ◽  
Base Line ◽  
Animal Reservoir ◽  
E Coli

coli is part of the normal gastrointestinal microbial flora of humans and animals. E. coli bacteria causing enteric/diarrhoeal disease are categorized into different groups based on their virulence properties and pathogenic features in humans. Enterohaemorrhagic E. coli (EHEC) are E. coli strains that  cause bloody diarrhoea  and haemolytic uraemic  syndrome  (HUS)  in humans, and have a defined zoonotic association. The major virulence factor of EHEC (and the actual  cause  of HUS)  is  the  ability  to  produce Shiga  toxins  (Stx),  thus  the  name Shiga Toxin Producing E. coli (STEC).  With enteropathogenic Escherichia  coli  (EPEC),  the diarrhoea in these  patients  is  due  to  attaching  and  effacing  (A/E)  lesions  in  the  enteric epithelium.    This risk assessment  was  conducted  after  a  human  outbreak  of  STEC  O103  in  2006, associated with contaminated dry-fermented sausages.   The Norwegian Scientific Committee for Food Safety (Vitenskapskomitéen for mattrygghet), Panel on  Biological  Hazards,  was  asked  by  the  Norwegian  Food  Safety  Authority (Mattilsynet) for  a  risk  assessment  regarding  shiga  toxin-producing  E.  coli  (STEC)  in  the Norwegian meat chain, with emphasis on dry-cured sausages. In response, an ad hoc Working Group  of  experts was  appointed with  the mandate  to  draft  a  risk  assessment  regarding  this issue.   The current report approaches the task by following and analysing the entire process, from the origin of the meats at farm level, to the final production and storage of dry-cured sausages. An overall  aim of  the  report has been  to  identify  and describe potential  intervention options  in various parts of this chain.     The main conclusions from the risk assessment are as follows:   It is  not  possible  to  give  any  reliable  quantitative  estimates  of  the  current  risk associated with consumption of dry-cured sausages.    There are  no  clear  indications  of  any  general  change  in  the  epidemiology  of  STEC  infections in humans in Norway over the last decade.   There is no documentation that there has been any change in the occurrence of various STEC in the domestic animal reservoir during the last decade.   The combination of proper slaughter hygiene and use of  thermal decontamination of sheep,  cattle  and  pig  carcasses  represents  an  efficient  way  to  reduce  STEC contamination. This approach would not only cause a reduction in the contamination level of STEC, but also provide a general beneficial effect on the level of other enteric pathogens, such as Salmonella and Yersinia enterocolitica.   Proper use  of  starter  cultures  in  fermentation,  combined  with  higher  fermentation temperatures,  will  reduce  the  probability  of  growth  of  STEC  in  contaminated  drycured sausages.   A combination  of  higher  fermentation  temperatures,  a  lower  pH  during  the  process, and heat-treatment of  the  final product should effectively eliminate  the potential  risk for transmission of STEC infections from consumption of dry-cured sausages. A 5 log reduction is possible.   Technological options  are  available  to  reduce  significantly  the  transfer  of  potential pathogens through meats in general, and specifically through dry-cured sausages.    The most important data gap is the lack of information about the actual occurrence of STEC infections  in  humans  in Norway.  Improved  laboratory  diagnostic  procedures and  epidemiological  surveillance,  combined with  better  reporting  and  tracing  in  the health care system are necessary.    The implementation  of  properly  designed  base-line  studies  of  various  domestic animals,  to  provide  data  on  the  occurrence  of  various  serotypes  and  their  virulence factors  present  is  recommended.  Also,  this  would  provide  a  better  basis  for comparison with human isolates. 

scholarly journals Precision long-read metagenomics sequencing for food safety by detection and assembly of Shiga toxin-producing Escherichia coli in irrigation water

2020 ◽  
Author(s):  
Meghan Maguire ◽  
Julie A. Kase ◽  
Dwayne Roberson ◽  
Tim Muruvanda ◽  
Eric W. Brown ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Shiga Toxin ◽  
Irrigation Water ◽  
Foodborne Illness ◽  
Nanopore Sequencing ◽  
E Coli ◽  
Antimicrobial Resistance Genes ◽  
Metagenomic Sample

ABSTRACTShiga toxin-producing Escherichia coli (STEC) contamination of agricultural water might be an important factor to recent foodborne illness and outbreaks involving leafy greens. Whole genome sequencing generation of closed bacterial genomes plays an important role in source tracking. We aimed to determine the limits of detection and classification of STECs by qPCR and nanopore sequencing using enriched irrigation water artificially contaminated with E. coli O157:H7 (EDL933). We determined the limit of STEC detection by qPCR to be 30 CFU/reaction, which is equivalent to 105 CFU/ml in the enrichment. By using Oxford Nanopore’s EPI2ME WIMP workflow and de novo assembly with Flye followed by taxon classification with a k-mer analysis software (Kraken), E. coli O157:H7 could be detected at 103 CFU/ml (68 reads) and a complete fragmented E. coli O157:H7 metagenome-assembled genome (MAG) was obtained at 105-108 CFU/ml. Using a custom script to extract the E. coli reads, a completely closed MAG was obtained at 107-108 CFU/ml and a complete, fragmented MAG was obtained at 105-106 CFU/ml. In silico virulence detection for E. coli MAGs for 105-108 CFU/ml showed that the virulotype was indistinguishable from the spiked E. coli O157:H7 strain. We further identified the bacterial species in the un-spiked enrichment, including antimicrobial resistance genes, which could have important implications to food safety. We propose this workflow could be used for detection and complete genomic characterization of STEC from a complex microbial sample and could be applied to determine the limit of detection and assembly of other foodborne bacterial pathogens.IMPORTANCEFoodborne illness caused by Shiga toxin-producing E. coli (STEC) ranges in severity from diarrhea to hemolytic uremic syndrome and produce-related incidence is increasing. The pervasive nature of E. coli requires not only detection, but also a complete genome to determine potential pathogenicity based on stx and eae genes, serotype, and other virulence factors. We have developed a pipeline to determine the limits of nanopore sequencing for STECs in a metagenomic sample. By utilizing the current qPCR in the FDA Bacteriological Analytical Manual (BAM) Chapter 4A, we can quantify the amount of STEC in the enrichment and then sequence and classify the STEC in less than half the time as current protocols that require a single isolate. These methods have wide implications for food safety, including decreased time to STEC identification during outbreaks, characterization of the microbial community, and the potential to use these methods to determine the limits for other foodborne pathogens.

scholarly journals Mixed Biofilm Formation by Shiga Toxin–Producing Escherichia coli and Salmonella enterica Serovar Typhimurium Enhanced Bacterial Resistance to Sanitization due to Extracellular Polymeric Substances†

2013 ◽  
Vol 76 (9) ◽  
pp. 1513-1522 ◽  
Author(s):  
RONG WANG ◽  
NORASAK KALCHAYANAND ◽  
JOHN W. SCHMIDT ◽  
DAYNA M. HARHAY
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Foodborne Pathogens ◽  
Shiga Toxin ◽  
Biofilm Formation ◽  
Extracellular Polymeric Substances ◽  
Bacterial Resistance ◽  
Salmonella Enterica Serovar Typhimurium ◽  
E Coli ◽  
Serovar Typhimurium

Shiga toxin–producing Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium are important foodborne pathogens capable of forming single-species biofilms or coexisting in multispecies biofilm communities. Bacterial biofilm cells are usually more resistant to sanitization than their planktonic counterparts, so these foodborne pathogens in biofilms pose a serious food safety concern. We investigated how the coexistence of E. coli O157:H7 and Salmonella Typhimurium strains would affect bacterial planktonic growth competition and mixed biofilm composition. Furthermore, we also investigated how mixed biofilm formation would affect bacterial resistance to common sanitizers. Salmonella Typhimurium strains were able to outcompete E. coli strains in the planktonic growth phase; however, mixed biofilm development was highly dependent upon companion strain properties in terms of the expression of bacterial extracellular polymeric substances (EPS), including curli fimbriae and exopolysaccharide cellulose. The EPS-producing strains with higher biofilm-forming abilities were able to establish themselves in mixed biofilms more efficiently. In comparison to single-strain biofilms, Salmonella or E. coli strains with negative EPS expression obtained significantly enhanced resistance to sanitization by forming mixed biofilms with an EPS-producing companion strain of the other species. These observations indicate that the bacterial EPS components not only enhance the sanitizer resistance of the EPS-producing strains but also render protections to their companion strains, regardless of species, in mixed biofilms. Our study highlights the potential risk of cross-contamination by multispecies biofilms in food safety and the need for increased attention to proper sanitization practices in food processing facilities.

scholarly journals Detection of Escherichia coli O157:H7 and Shiga toxin 2a gene in pork, pig feces, and clean water at Jagalan slaughterhouse in Surakarta, Central Java Province, Indonesia

2019 ◽  
Vol 12 (10) ◽  
pp. 1584-1590
Author(s):  
Maria Kristiani Epi Goma ◽  
Alvita Indraswari ◽  
Aris Haryanto ◽  
Dyah Ayu Widiasih
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Shiga Toxin ◽  
Water Samples ◽  
Animal Origin ◽  
Clean Water ◽  
Escherichia Coli O157 ◽  
Fecal Samples ◽  
E Coli ◽  
Pork Samples

Background and Aim: The feasibility assessment of food products on the market becomes one of the milestones of food safety. The quality of food safety of animal origin especially pork need to get attention and more real action from the parties related and concerned. Since pork is also a source of transmission for the contagion of foodborne disease so that the study of the existence of several agents in the pork and its products become the benchmark of safety level. This study aimed to isolate, identify, and detect the Shiga toxin 2a (stx2a) gene from Escherichia coli O157:H7 in pork, pig feces, and clean water in the Jagalan slaughterhouse. Materials and Methods: A total of 70 samples consisting of 32 pork samples, 32 pig fecal samples, and 6 clean water samples were used to isolate and identify E. coli O157:H7 and the stx2a gene. Isolation and identification of E. coli O157:H7 were performed using culture on eosin methylene blue agar and Sorbitol-MacConkey agar media and confirmed molecularly with polymerase chain reaction to amplify the target genes rfbE (317 bp) and fliC (381 bp). The isolates, which were identified as E. coli O157:H7, were investigated for the stx2a gene (553 bp). Results: The results of this study show that of the total collected samples, E. coli O157:H7 was 28.6% in Jagalan slaughterhouse and consisted of 25% of pork samples, 31.25% of pig fecal samples, and 33.3% of clean water samples. The isolates that were identified to be E. coli O157:H7 mostly contained the stx2a gene, which was equal to 75%, and consisted of seven isolates from pork samples, seven isolates from fecal samples, and one isolate from clean water samples. Conclusion: E. coli O157:H7 was found in 28.6% of pork, pig feces, and clean water in Jagalan slaughterhouse and 75% of identified E. coli O157:H7 contained the stx2a gene.

scholarly journals Characterization of Clinical Escherichia coli Strains Producing a Novel Shiga Toxin 2 Subtype in Sweden and Denmark

2021 ◽  
Vol 9 (11) ◽  
pp. 2374
Author(s):  
Xiangning Bai ◽  
Flemming Scheutz ◽  
Henrik Mellström Dahlgren ◽  
Ingela Hedenström ◽  
Cecilia Jernberg
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Virulence Genes ◽  
Epidemiological Surveillance ◽  
E Coli ◽  
Shiga Toxin 2 ◽  
Life Threatening ◽  
Macrolide Resistance Gene

Shiga toxin (Stx) is the key virulence factor in the Shiga Toxin-Producing Escherichia coli (STEC), which can cause diarrhea and hemorrhagic colitis with potential life-threatening complications. There are two major types of Stx: Stx1 and Stx2. Several Stx1/Stx2 subtypes have been identified in E. coli, varying in sequences, toxicity and host specificity. Here, we report a novel Stx2 subtype (designated Stx2m) from three clinical E. coli strains isolated from diarrheal patients and asymptomatic carriers in Sweden and Denmark. The Stx2m toxin was functional and exhibited cytotoxicity in vitro. The two Swedish Stx2m-producing strains belonged to the same serotype O148:H39 and Multilocus Sequencing Typing (MLST) Sequence Type (ST) 5825, while the Danish strain belonged to the O96:H19 serotype and ST99 type. Whole-genome sequencing (WGS) data analysis revealed that the three Stx2m-producing strains harbored additional virulence genes and the macrolide resistance gene mdf (A). Our findings expand the pool of Stx2 subtypes and highlight the clinical significance of emerging STEC variants. Given the clinical relevance of the Stx2m-producing strains, we propose to include Stx2m in epidemiological surveillance of STEC infections and clinical diagnosis.

scholarly journals Dual-Serotype Biofilm Formation by Shiga Toxin-Producing Escherichia coli O157:H7 and O26:H11 Strains

2012 ◽  
Vol 78 (17) ◽  
pp. 6341-6344 ◽  
Author(s):  
Rong Wang ◽  
Norasak Kalchayanand ◽  
James L. Bono ◽  
John W. Schmidt ◽  
Joseph M. Bosilevac
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Potential Risk ◽  
Shiga Toxin ◽  
Biofilm Formation ◽  
Escherichia Coli O157 ◽  
Content Type ◽  
E Coli

ABSTRACTEscherichia coliO26:H11 strains were able to outgrow O157:H7 companion strains in planktonic and biofilm phases and also to effectively compete with precolonized O157:H7 cells to establish themselves in mixed biofilms.E. coliO157:H7 strains were unable to displace preformed O26:H11 biofilms. Therefore,E. coliO26:H11 remains a potential risk in food safety.

scholarly journals Prevalence and Characterization of Extended-Spectrum β-Lactamase-Producing Antibiotic-Resistant Escherichia coli and Klebsiella pneumoniae in Ready-to-Eat Street Foods

Antibiotics ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 850
Author(s):  
Shobha Giri ◽  
Vaishnavi Kudva ◽  
Kalidas Shetty ◽  
Veena Shetty
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Klebsiella Pneumoniae ◽  
Rural Areas ◽  
Significant Risk ◽  
Microbiological Quality ◽  
Resistant Bacteria ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Extended Spectrum

As the global urban populations increase with rapid migration from rural areas, ready-to-eat (RTE) street foods are posing food safety challenges where street foods are prepared with less structured food safety guidelines in small and roadside outlets. The increased presence of extended-spectrum-β-lactamase (ESBL) producing bacteria in street foods is a significant risk for human health because of its epidemiological significance. Escherichia coli and Klebsiella pneumoniae have become important and dangerous foodborne pathogens globally for their relevance to antibiotic resistance. The present study was undertaken to evaluate the potential burden of antibiotic-resistant E. coli and K. pneumoniae contaminating RTE street foods and to assess the microbiological quality of foods in a typical emerging and growing urban suburb of India where RTE street foods are rapidly establishing with public health implications. A total of 100 RTE food samples were collected of which, 22.88% were E. coli and 27.12% K. pneumoniae. The prevalence of ESBL-producing E. coli and K. pneumoniae was 25.42%, isolated mostly from chutneys, salads, paani puri, and chicken. Antimicrobial resistance was observed towards cefepime (72.9%), imipenem (55.9%), cefotaxime (52.5%), and meropenem (16.9%) with 86.44% of the isolates with MAR index above 0.22. Among β-lactamase encoding genes, blaTEM (40.68%) was the most prevalent followed by blaCTX (32.20%) and blaSHV (10.17%). blaNDM gene was detected in 20.34% of the isolates. This study indicated that contaminated RTE street foods present health risks to consumers and there is a high potential of transferring multi-drug-resistant bacteria from foods to humans and from person to person as pathogens or as commensal residents of the human gut leading to challenges for subsequent therapeutic treatments.

scholarly journals Prevalence and molecular characterisation of shiga-toxin-producing Escherichia coli in raw milk cheeses from Lazio region, Italy

2016 ◽  
Vol 5 (1) ◽  
Author(s):  
Selene Marozzi ◽  
Paola De Santis ◽  
Sarah Lovari ◽  
Roberto Condoleo ◽  
Stefano Bilei ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Raw Milk ◽  
Foodborne Diseases ◽  
Control Plan ◽  
Lazio Region ◽  
Standard Methods ◽  
E Coli ◽  
Regional Control ◽  
Unpasteurized Milk

In recent years, the incidence of foodborne diseases caused by shiga toxin-producing <em>Escherichia coli</em> (STEC) has increased globally. For this reason, within the specific regional control plan for the detection of STEC in food products in Italy, the presence of STEC in unpasteurized milk cheeses was investigated. In total 203 samples obtained from March 2011 to December 2013 were analysed, with two standard methods (ISO 16654:2001 and ISO 13136:2012). Two strains of <em>E. coli</em> O157 were isolated (2/161, 1.2%) but did not carry any virulence-associated genes and 22 <em>stx</em>-positive samples (22/146, 15.1%) were detected in enrichment cultures, mostly from ovine cheeses. Only two strains isolated from different ovine cheeses carried <em>stx</em> gene and none of these was <em>eae</em>-positive. This study confirms the presence of <em>stx</em>-positive <em>E. coli</em> and suggests that this type of food cannot be excluded as a potential vehicle of STEC.

scholarly journals Novel Hybrid of Typical Enteropathogenic Escherichia coli and Shiga-Toxin-Producing E. coli (tEPEC/STEC) Emerging From Pet Birds

2018 ◽  
Vol 9 ◽  
Author(s):  
Rosely Martins Gioia-Di Chiacchio ◽  
Marcos Paulo Vieira Cunha ◽  
Lilian Rose Marques de Sá ◽  
Yamê Minieiro Davies ◽  
Camila Bueno Pacheco Pereira ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Enteropathogenic Escherichia Coli ◽  
E Coli ◽  
Pet Birds

Introduction to the Food Safety Concerns of Verotoxin-Producing Escherichia coli

2003 ◽  
Vol 228 (4) ◽  
pp. 331-332 ◽  
Author(s):  
Hussein S. Hussein ◽  
Stanley T. Omaye
Keyword(s):  
Escherichia Coli ◽  
Food Safety ◽  
Safety Concern ◽  
Critical Evaluation ◽  
The United States ◽  
Global Perspective ◽  
E Coli ◽  
The Past ◽  
Food Borne Pathogens ◽  
Food Borne

Verotoxin-producing Escherichia coli (VTEC) have emerged in the past two decades as food-borne pathogens that can cause major outbreaks of human illnesses worldwide. The number of outbreaks has increased in recent years due to changes in food production and processing systems, eating habits, microbial adaptation, and methods of VTEC transmission. The human illnesses range from mild diarrhea to hemolytic uremic syndrome (HUS) that can lead to death. The VTEC outbreaks have been attributed to O157:H7 and non-O157:H7 serotypes of E. coli. These E. coli serotypes include motile (e.g., O26:H11 and O104:H21) and nonmotile (e.g., O111:H–,0145:H–, and O157:H–) strains. In the United States, E. coli O157:H7 has been the major cause of VTEC outbreaks. Worldwide, however, non-O157:H7 VTEC (e.g., members of the 026, O103, O111, O118, O145, and O166 serogroups) have caused approximately 30% of the HUS cases in the past decade. Because large numbers of the VTEC outbreaks have been attributed to consumption of ruminant products (e.g., ground beef), cattle and sheep are considered reservoirs of these food-borne pathogens. Because of the food safety concern of VTEC, a global perspective on this problem is addressed (Exp Biol Med Vol. 228, No. 4). The first objective was to evaluate the known non-O157:H7 VTEC strains and the limitations associated with their detection and characterization. The second objective was to identify the VTEC serotypes associated with outbreaks of human illnesses and to provide critical evaluation of their virulence. The third objective was to determine the rumen effect on survival of E. coli O157:H7 as a VTEC model. The fourth objective was to explore the role of intimins in promoting attaching and effacing lesions in humans. Finally, the ability of VTEC to cause persistent infections in cattle was evaluated.

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.

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