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E. coli: Whats next?. HOST Bill McDowell Editorial Director, Meatingplace MODERATOR Mike Fielding Editor, Meatingplace.

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Presentation on theme: "E. coli: Whats next?. HOST Bill McDowell Editorial Director, Meatingplace MODERATOR Mike Fielding Editor, Meatingplace."— Presentation transcript:

1 E. coli: Whats next?

2 HOST Bill McDowell Editorial Director, Meatingplace MODERATOR Mike Fielding Editor, Meatingplace

3 POLL QUESTIONS Will the Big Six be declared to be product adulterants? Yes No Should they be? Yes No

4 Non-O157 Shiga Toxin Producing E. coli: Biology, Detection and Significance Mohammad Koohmaraie, Ph.D. CEO – Meat Division IEH Laboratories & Consulting Group Lake Forest Park, WA

5 Presentation Outline Nomenclature O157 vs. non-O157 Will Non-O157 STECs get adulterant designation? Detection – FSIS Approach Prevalence Summary and conclusions

6 Nomenclature E. coli O157:H7

7 Y Y Lipopolysaccharide (LPS) Flagella = H antigen = O antigen H1-H56 O1-O173 E. coli Serotyping O157:H7 O104:H4 O26:H11

8 Pathogenicity Not all E. coli strains are pathogenic. To be pathogenic, a strain must have the necessary properties to cause disease in human. These properties are called virulence factors. Exactly what virulence factors are required is unknown.

9 Pathogenicity E. coli can cause human disease when they possess stx1 or stx2. Individuals infected with strains producing stx2 are more likely to develop severe disease than those infected with strains carrying Shiga toxin 1. It is commonly thought that E. coli must contain stx1 or stx2 and eae (intimin) and its substitutes to have the highest chance of causing disease in humans – of course there are always exceptions.

10 S higa T oxin producing E scherichia C oli STEC E ntro Hemorrhagic E scherichia C oli EHEC pSTEC

11 Top Non-O157 Serotypes (CDC) – O2622% of non-O157 STEC – O11116% of non-O157 STEC – O10312% of non-O157 STEC – O121 9% of non-O157 STEC – O45 7% of non-O157 STEC – O145 5% of non-O157 STEC

12 Annual Submission of Non-O157 to PulseNet CDC

13 E. coli O157 vs. Non-O157 E. coli

14 E. coli O157 ingested days non-bloody diarrhea, abdominal cramps days resolution 92%8% HUS bloody diarrhea days 80% Mead. Lancet 1998 Sequence of events in E. coli O157:H7 infection CDC

15 Non-O157 STEC ingested days days resolution 99% bloody diarrhea days 40% non-bloody diarrhea, abdominal cramps <1% HUS Sequence of events in Non-O157 infection CDC

16 Food commodities implicated outbreaks of non-O157 STEC infections with known food commodity, CommodityNumber outbreaksFood items Fruit-nuts3Juice, apple cider, berries Dairy2Cheese, margarine Leafy vegetables1Lettuce No outbreaks due to beef

17 But it all changed in 2010 with the recall of ground beef because of human illness associated with O26 contamination

18 Will Non-O157 STECs Get Adulterant Designation? It is a matter of when and not if any more Less than 12 months

19 Detection

20 Testing for STEC E. coli O157 – Unusual feature: does not ferment sorbitol Non-O157 STEC – Lack unusual features – A group of organisms

21 USDA Approach Meat sample Enrich Screen for eae + stx If negative – STOP If positive, PCR to determine if it is a top 6 If one of the top 6 – Sample is POSITIVE If not a top 6 – Sample is NEGATIVE

22 Non-O157 Prevalence

23 Prevalence of Non-O157 STEC Commercial fed cattle processing plants Commercial fed cattle processing plants as a function of the season of the year Commercial cow/bull processing plants Commercial lamb processing plants Imported raw ground beef material (trim) National ground beef supply We are very appreciative of the U.S. meat industry for allowing us to use their facilities as our laboratory.

24 Results Pre-evisceration (No intervention) Final (after all interventions) E. coli O % (144/324 carcasses) 1.8% (6/326 carcasses) Non-O157 STEC 54% (180/334 carcasses) 8.3% (27/326 carcasses)

25 STEC Prevalence in Imported and Domestic Boneless Beef Trim Used for Ground Beef

26 STEC Frequency of STEC isolation in boneless beef trim by country of origin

27 Serotypes of STEC isolated by country Underlined serotypes have been associated with human illness. Bolded serotypes have been associated with Hemolytic Uremic Syndrome (HUS).

28 A National Survey of the Prevalence of non-O157 Shiga toxin-producing E. coli in Ground Beef

29 BIFSCo Database Microbiological Regions

30 Ground Beef Non-O157 STEC 4133 samples Supplies by ground beef manufacturers over a 24 months period 24.3% (1006/4133) of the samples were positive for Shiga toxin (stx1 or stx2). In all, 0.2% of the samples (10/4133) could be considered a food safety threat.

31 Control Hide is the principal source of most pathogens. An effective control in order of significance is: 1 st - Dressing practices 2 nd - Effective interventions Effective – scientific support Works in your hands – validation Must make contact the potential pathogens 3 rd - Effective test and hold

32 Summary and Conclusions STEC are a natural part of the animal microflora. Some Non-O157 STEC can cause severe disease in humans. Non-O157 STEC is found at high frequency in pre- harvest samples (feces and hides). The prevalence on carcass depends on the adequacy of dressing practices.

33 Summary and Conclusions A thoughtful and comprehensive approach should be used to develop a national policy with respect to the control of non-O157 STEC. This approach should be such that remove the most virulent E. coli from the food supply not just some and not just any. In this way we are truly reducing the burden to public health of pathogenic E. coli without undue economic hardship.

34 Mike Mullen Corporate Account Manager For more information contact: or

35 The Sinful Six Interventions to Minimize the Risk for Beef Processors: A Research and Application Update Harshavardhan Thippareddi, Ph.D. Associate Professor and Extension Food Safety Microbiologist Dept. of Food Science and Technology University of Nebraska, Lincoln, NE 68583

36 Outline Introduction Prevalence of STECs Antimicrobial Interventions for STECs Non-O157 STEC Challenges for the processor Non-intact Beef – STECs Summary

37 Innovative Interventions Developed and Applied by the Beef Industry 1993 E. coli O157:H7 Pacific Northwest – Knife trimming and water washing 2009 – Multiple meat processing interventions – Sequential – Hurdle Technologies – Primary and Secondary – At all phases of meat processing

38 Outline Introduction Prevalence of STECs Antimicrobial Interventions for STECs Non-O157 STEC Challenges for the processor Non-intact Beef – STECs Summary

39 Cattle STEC prevalence data varies: – 0-19%, dairy cattle 1,2,3,4 – 19.4 – 56.3% 5, beef cattle feces/hides Food prevalence data very limited: – Pre-evisceration beef carcasses >50% 5,6 – Retail ground beef, 2.3% 7 1 Wachsmuth et al., Wells et al., Cray et al., Thran et al., Barkocy-Gallagher, et al., Arthur et al., Samadpour et al., 2006 Dr. Hagen, 2007 Non-O157 Shiga toxin producing E. coli – From FSIS Public Meeting, 2007

40 Outline Introduction Prevalence of STECs Antimicrobial Interventions for STECs Non-O157 STEC Challenges for the processor Non-intact Beef – STECs Summary

41 Antimicrobial Interventions for Slaughter, Fabrication and Grinding

42 Antimicrobial Agents: Classification Direct Food Additives Sod. or Pot. Lactates, Buffered sodium citrate, sod. Diacetate and Lactoferrin, Irradiation Considered ingredients, need to be labeled as such Secondary Direct Food Additives Peroxy acids, ASC, Ozone No labeling requirement

43 Slaughter: – Chemical dehairing – Hot water rinses – Steam pasteurization – Steam vacuum – Chemical rinses – Lactoferrin Antimicrobial Interventions

44 Fabrication: – Organic acid rinses – Sanova – Ozone – Per-acetic acid – Lauric Arginate – Lactoferrin Antimicrobial Interventions

45 Trim for Grinding: – Organic acid rinses – Ozone – Per-acetic acid – ASC – Multiple hurdle technology – High Pressure Processing Ground Beef: – Irradiation Antimicrobial Interventions

46 Chlorine, Organic acids most commonly used – Organic acids – lactic, acetic, citric Improves microbiological quality of carcasses Other chemicals include Per-oxy acetic acid, Acidified sodium chlorite, CPC Chemical Rinses

47 Antimicrobials - Variety High InoculumLow Inoculum SurvivalReductionSurvivalReduction Non-Trt AC, 0.001% AA, 2% LA, LB, 1% PAA, 0.02% ASC, 0.02% CPC, 0.5% Ransom et al., 2003 Boneless beef short plates & Lean tissue pieces; E. coli O157:H7

48 Antimicrobial Interventions for Non- intact Beef Oklahoma State / Ross (2010) study compared 10 different antimicrobials for efficacy on surface inoculated beef subprimals CytoGuard is a registered trademark of A&B Ingredients Courtesy: Arun Ramabadran, Spray Systems Co., Wheaton, IL

49 Outline Introduction Prevalence of STECs Antimicrobial Interventions for STECs Non-O157 STEC Challenges for the processor Non-intact Beef – STECs Summary

50 Non-O157 STEC Challenges for the Beef Processor Biology and characteristics of the organisms are unknown Characteristics of Significance – Thermal resistance – Acid resistance – Dehydration – Aw – Irradiation – Behavior to antimicrobial interventions traditionally used in beef processing

51 Characteristics of Significance in Meat Processing – 10°C Strain O26:H11 survived, but did not grow LPD h to 39.4 h Growth rate – to log CFU/g/h O145 and O157 had shorter LPD compared to O45, O103, and O121 (O111 had intermediate LPD) After end of LPD, no difference in growth rate (Chatzikyriakidiu et al., 2011)

52 Characteristics of Significance in Meat Processing – Growth (Thippareddi et al., Unpublished)

53 Characteristics of Significance in Meat Processing – Thermal Resistance L. monocytogenesSalmonella spp. LeanFattyLeanFatty 125°F °F °F Z Value STECsBrothGround Beef 130°F (54.4°C) °F (60°C) °F (65.5°C) (Fain et al., 2009; Leong et al., 2011, Vasan et al., 2011)

54 Characteristics of Significance in Meat Processing – Acid Resistance E. coli O157:H7 reductions of >7.70 log CFU/ml was observed in lemon and lime juices* within 72 h at 22°C STECs (six serotypes) reduction of >6 log within 72 h at 22°C in lemon and lime juices (Enache et al., 2009; Kataoka et al., 2011) *Lemon Juice: 6.8° Brix; 2.62 pH; 4.62 TA *Lime Juice: 7.4° Brix; 2.54 pH; 4.82 TA

55 Non-O157 STECs – Properties of Significance, Acid Resistance Montet et al., 2009

56 Outline Introduction Prevalence of STECs Antimicrobial Interventions for STECs Non-O157 STEC Challenges for the processor Non-intact Beef – STECs Summary

57 Translocation and Thermal Inactivation of Shiga Toxin- Producing E. coli in Blade and Chemically Tenderized Beef 1.Luchansky, J. B., J. E. Call, R. K. Phebus, and H. Thippareddi Translocation of surface inoculated Escherichia coli O157:H7 into beef subprimals following blade tenderization. J. Food Prot. 71: Luchansky, J. B., A. C. S. Porto-Fett, B. Shoyer, R. K. Phebus, H. Thippareddi, and J. E. Call Thermal inactivation of Escherichia coli O157:H7 in blade tenderized beef steaks cooked on a commercial open- flame gas grill. J. Food Prot. 72: Luchansky, J. B., A. C. S. Porto-Fett, B. A. Shoyer, J. E. Call, W. Schlosser, W. Shaw, N. Bauer, and H. Latimer Inactivation of Shiga toxin-producing O157:H7 and non-O157:H7 Shiga toxin-producing Escherichia coli in brine-injected, gas-grilled steaks. J. Food Prot. 74: Luchansky, J. B., A. C. S. Porto-Fett, B. A. Shoyer, J. E. Call, W. Schlosser, W. Shaw, N. Bauer, and H. Latimer Fate of Shiga toxin-producing O157:H7 and non-O157:H7 Shiga toxin-producing Escherichia coli in blade tenderized beef steaks cooked on a commercial open-flame gas grill. J. Food Prot., Submitted

58 STEC and ECOH behave similarly! No discernible differences in translocation between ECOH and STEC following blade tenderization or chemical injection of beef sub-primals – Majority of cells to top-most 1 cm – More transfer via single pass and lean side inoculation and tenderization than double pass and fat side inoculation and tenderization (NCBA Study) Courtesy: Dr. John Luchansky & the ARS Non-Intact Team

59 STEC and ECOH behave similarly! No discernible differences in thermal resistance between STEC and ECOH following cooking of blade tenderized or chemically-injected steaks – Higher temperatures generated greater lethality – No difference in lethality based on steak thickness (1.0 or 1.5 in.) – Subtle differences in thermal resistance between steaks injected with lactate-containing brine compared to brine without lactate that were cooked at 140 F Courtesy: Dr. John Luchansky & the ARS Non-Intact Team

60 Outline Introduction Prevalence of STECs Antimicrobial Interventions for STECs Non-O157 STEC Challenges for the processor Non-intact Beef – STECs Summary

61 STECs seem to be similar in characteristics to E. coli O157:H7 based on current research Very few interventions eliminate the E. coli O157:H7 and non-O157 STEC risk Novel technologies such as carcass irradiation and HPP should be optimized and validated to assure safety of intact as well as non-intact beef products

62 The beef industry has made significant strides in reducing risks and costs associated with foodborne pathogens. – However, food safety remains a significant concern due to increasing media and regulatory scrutiny. – E. coli O157 is naturally occurring in animals and the environment, which makes it an ever-present challenge. – The beef industry has a good track record for safety, but there is always room to improve. Now there is a tool available to your suppliers that can help reduce E. coli O157 before it enters the packing plant in the cattle, at the feedlot thereby helping reduce E. coli O157 throughout the beef processing chain. – Escherichia Coli Bacterial Extract vaccine* with SRP ® technology is the only conditionally licensed product that can help reduce E. coli O157** carrier state prevalence and shedding at the source in the cattle. – Reducing the amount of E. coli O157 coming into the plant can give other interventions in place an opportunity to be as effective as possible. The goal for us at Pfizer Animal Health is to help ensure continued confidence in the industry to help keep demand for our products strong. * This product license is conditional. Efficacy and potency test studies are in progress. ** Escherichia coli O157:H7 bacterial strain 1 Woerner DR, Ransom JR, Sofos JN, Dewell GA, Smith GC, Salman MD, et al. Determining the Prevalence of Escherichia coli O157 in Cattle and Beef from the Feedlot to the Cooler. J Food Prot 2006;69: Together, we can reduce E. coli O157. Dr. Brad Morgan, Food Safety Specialist Pfizer Animal Health

63 E.COLI: Legal Issues and the Big Six Robert G. Hibbert K&L Gates LLP

64 E.coli 0157:H7 Precedent – 1994 Policy Statement – Texas Food Industry v. Espy (1994) – Extension to other products – trim, non-intact – Avoidance of rulemaking – Integration with HACCP

65 E.coli Precedent (continued) – Commercial response – Technological responses – Exception proves the rule-labeling and product handling distinctions

66 Shiga–Toxin Producing E.Coli Serotypes – Outbreaks – Methodology issues – Marler petition – Pending policy statement

67 Legal Issues (Regulatory) – Adulterants under 1(m)(1) vs. 1 (m)(3) – Policy by Recall – Abandonment of rulemaking (listeria RTE precedent) – HACCP and inspection implementation

68 Legal Issues (Commercial) – Guarantees and requirements – Role of testing – Reliability of testing – Tort liability – Insurance concerns

69 QUESTIONS & ANSWERS

70 FOR MORE INFORMATION Mohammad Koohmaraie, Ph.D.: Harshavardhan Thippareddi, Ph.D.: Robert G. Hibbert: Mike Fielding: Bill McDowell: Ecolab: Pfizer Animal Health: Webinar recording and PowerPoint presentation will be ed to you within 48 hours. For more information:


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