1. UEHA 2013 Special Workshop

2. Topic15 minute exerciseFood microbiology 1 refresherContact APCFoodborne illness 2 overviewpH and Aw testing3 STECs and SalmonellaColiform plating on petrifilms4L. monocytogenes and Campylobacter5 Norovirus and Hepatitis ACheese sampling S. aureus carrier sampling 3. 1. 2. 3. 4.Label 2 agar plates with your name. Press three or four fingertips to one agar plate. Kiss the second agar plate lightly. Well collect these plates and incubate them, then provide pictures of the results. 4. Topic15 minute exerciseFood microbiology 1 refresherContact APCFoodborne illness 2 overviewpH and Aw testing3 STECs and SalmonellaColiform plating on petrifilms4L. monocytogenes and Campylobacter5 Norovirus and Hepatitis ACheese sampling S. aureus carrier sampling 5. 1 in 3 1 in 51 in 6 1 in 12 6. Annual foodborne illnesses 2011 Norovirus5,461,731 Yersinia enterocoliticaSalmonella spp1,027,561 Unicellular parasitesClostridium perfringens965,958 VibriosCampylobacter845,024 Listeria monocytogenesStaphylococcus aureus241,148 TrichinellaSTEC175,905 C. botulinumShigella131,25497,752 ~70,00052,408 1591 156 55 7. 50 6003,000 40,000 8. Annual foodborne illness deaths NorovirusSalmonella spp Clostridium perfringens Campylobacter Staphylococcus aureus STEC Shigella149 Yersinia enterocolitica 378 Unicellular parasites 26 Vibrios 76 Listeria monocytogenes 6 Trichinella 20 C. botulinum 1029327 48 255 0 9 9. SalmonellosisBotulism Cholera / Typhoid FeverNorovirus Gastroenteritis 10. SalmonellosisBotulism Cholera / Typhoid FeverNorovirus Gastroenteritis 11. Meats PoultryEggs All answers are correct 12. 33 25>250 2,033 13. 4% 40%14% 44% 14. 2% 12%18% 21% 15. Other/Multiple 2%Unknown 29% Norovirus 40%Chemical 5% Parasites 1% Bacteria 23% 16. True False 17. Listeria monocytogenes Norovirus Salmonella Clostridium botulinum 18. Very young Very oldInfirm (sick) Allergic individuals 19. Stomach acids Immune System Intestinal flora competition Just pure luck 20. Where a pathogens toxin causes illnessWhere a parasite invades host tissue Where a pathogen causes direct illness by invading host tissue Where a pathogen causes direct illness by attaching to or invading host tissue 21. Global food sources Consumer choice for fresh foodsEmerging pathogens Lack of on-farm food safety 22. Foodborne illnesses Norovirus1Yersinia enterocolitica8Salmonella spp2Unicellular parasites9Clostridium perfringens3Vibrios10Campylobacter4Listeria monocytogenes11Staphylococcus aureus5Trichinella12STEC6C. botulinum13Shigella7Non-infectious14 23. 1. 2. 3. 4.Label 2 agar plates with your name. Press four fingertips to one agar plate. Kiss the second agar plate lightly. Well collect these plates and incubate them, then provide pictures of the results. 24. Topic15 minute exerciseFood microbiology 1 refresherContact APCFoodborne illness 2 overviewpH and Aw testing3 STECs and SalmonellaColiform plating on petrifilms4L. monocytogenes and Campylobacter5 Norovirus and Hepatitis ACheese sampling S. aureus carrier sampling 25. 1. Remove 5 ml of milk from sample container and pipet onto coliform petrifilm. 2. Press petrifilm with plastic tool. 3. Well collect these films, incubate them, then provide pictures of the results. 26. E. Coli O157: H7 CONTENTS 27. 1.ETEC Enterotoxigenic -Travellers diarrhea a.2.3.4.Has toxins that cause diarrheaEPEC Enteropathogenic attaches to intestinal walls causing diarrhea and malabsorption (mostly in infants) EIEC Enteroinvasive attaches, then invades intestinal mucosa causing dysentery (bloody diarrhea). Most likely plasmid from Shigella STEC/EHEC - Shiga Toxin Enterohemorrhagic (discussed below) 28. 1. Typical STEC EHEC outbreakFecal origins of E. coliSome form of cross contamination occurs Either food is consumed raw or undercooked Person eats the foodIllness starts in 3-4 days, but can be 1-9 days 29. Mild Hemorrhagic colitis (2-9 days) Asymptomatic to mild diarrheaAbdominal cramps and dysenteryHemolytic uremic syndrome (3-7%) thrombotic thrombocytopenia purpura Internalized cells with Stx2 toxin in blood attach to and attack kidney cells 3-5% HUS mortality 30. G-, non-sporeformer, facultative 400 serotypes of STEC only a few EHEC O157 predominant strain (75%) O111, O26, O121, O103, O145, and O45 are others Coliform assay usually negative (O157H7) Survives difficult environments (dry and acid) Virulence associated with pathogenic 31. Shiga toxin 1 (Stx1) is nearly identical to the toxin produced by Shigella dysenteriae Type I. Most likely genetically transferred by plasmids Shiga toxin 2 (Stx2) - is most often associated with severe sequelae, such as hemolytic uremic syndrome (HUS), characterized by acute renal failure. LEE (locus for enterocyte effacement; intimin, a protein that enables bacterial attachment to epithelial cells). 32. USA 175,905 est. STEC illnesses annually Approx 63,000 are EHEC 20 deaths annually 33. Foods subjected to fecal-oral contaminationUndercooked hamburger (1982 Jack-in-box outbreak)Unpasteurized juices (fecal contamination from drops) yogurt, mayonnaise, fermented sausage, cheese (note pH below 4.6 EC is acid tolerant) fresh produce (lettuce, spinach, etc) 34. CookingPrevent cross contamination 35. Consumer education 36. CONTENTS 37. 1. Typical outbreakSalmonella is a classic fecal contaminateSome form of cross contamination occursProcessing fails to kill all SalmonellaPerson eats the foodIllness starts in 6-72 hours 38. Common Diarrhea & abdominal cramps Fever Additional symptoms: Bloody diarrhea Vomiting Headache Body aches Reactive arthritis 39. G-, non-sporeformer, facultative, rod Survives difficult environments (dry and acid) Predominantly fecal S. enterica Enteritidis and S. enterica Typhimurium >2500 serotypesRequires large numbers for infection OR only 1 if protected from stomach acids 40. An adhesion protein (attaches cells to intestinal walls) An invasion protein (internalizes cells) Other survival factors can determine severity and length of illness Mostly genomic encoded dont see high levels of DNA transfer 41. USA 1,027,561 591 est. illnesses/yr Second largest number afterNorovirus 378 deaths annually Deaths in immunocompromised Use PFGE to quickly determine serovar and strain Used to implicate multistate outbreaks 42. meats,poultry, eggs, milk and dairy products, fish, shrimp, spices, yeast, coconut, sauces, unpasteurized salad dressings, cake mixes, cream-filled desserts and toppings that contain raw egg, dried gelatin, peanut butter, cocoa, produce (fruits and vegetables, such as tomatoes, peppers, and cantaloupes), and chocolate. 43. Proper cookingPrevent cross contamination 44. Sanitation and GMPs Prevent ingress or spread of Salmonella Enhance the stringency of hygiene practices Apply hygienic design principles to building and equipment design Prevent or minimize growth of Salmonella within the facility Establish a raw materials/ingredients control program Validate control measures to inactivate Salmonella Establish procedures for verification of Salmonella controls and corrective actions 45. Consumer education 46. Topic15 minute exerciseFood microbiology 1 refresherContact APCFoodborne illness 2 overviewpH and Aw testing3 STECs and SalmonellaColiform plating on petrifilms4L. monocytogenes and Campylobacter5 Norovirus and Hepatitis ACheese sampling S. aureus carrier sampling 47. CONTENTS 48. 1. Typical outbreakListeria monocytogenes found in food processing environment Some form of cross contamination occurs Lm can grow slowly in foods even under refrigeration Person eats the foodIllness starts in >12 hours, but can be several days 49. Gastrointestinal Mild flu-like symptoms (persistentfever, mild nausea, mild vomiting) Invasive (enters phagocytic immune cells) Septicemia Meningitis Spontaneous abortion barrier)(crosses placental 50. G+, non-sporeformer, facultative Grows under refrigeration (slowly) Survives difficult environments Isolated from 1-10% human feces, many animals, soil, water, and sewage Often found in food processing 51. Many different serovars one highly virulent (4b) while others not at allRequires no.s for infection due to success of human immune system Virulence associated with pathogenic factors (genes 52. An adhesin protein (attaches cells to intestinal walls) Internalin encourages immune cells to phagocytize LM Listeriolysin O protein that helps LM escape immune cells vacuoles to survive in cytoplasm ActA helps form listiopods (protrusions of cells inside immune cell that are engulfed by a neighboring cell (transmits LM from cell to cell) 53. USA 1591 est. illnesses annually 255 deaths annually Highest FBI death rate (~16%) 54. Foodsthat require considerable handling that are either minimally process (fresh/raw) or are handled extensively AFTER thermal processing or other lethality's meat and meat products (hot dogs, deli meats) Raw milk and dairy products (raw milk cheese) fresh produce (cantaloupe 2012) 55. SanitationPrevent cross contamination 56. Anti-listerial additivesPost packaging thermal processing 57. Consumer education 58. Campylobacter CONTENTS 59. 1. Typical outbreakFecal origins in poultry, swine, sheep and cattle Some form of cross contamination occurs Either food is consumed raw or undercooked Person eats the foodIllness starts in 2-5 days 60. Gastrointestinal (2-10 days) Bacteremia (0.15% progress) Fever, diarrhea, cramps, vomiting May have muscle aches, nausea, headacheMeningitis, hepatitis, pancreatitisGuillan barre (0.1% of gastroenteritis) Autoimmune disease Immune system created to attack Campy attacks certain neurological cells by mistake About 40% of GB have had Campybacteriosis 61. G-, non-sporeformer, S-shaped C. jejuni major pathogenic species Microaerophilic (prefer O2 3-5%) Difficult to culture susceptible to drying, heating, freezing, disinfectants, and acidic conditions Takes ~10,000 cells for infection 62. Adhesion proteins Invasion proteins Toxin proteins CDT cytolethal distending toxin Hemolysin toxin 63. USA 845,024 est. illnesses annually Most undiagnosed Most in very young 1-12 months and other immunocompromised 76 deaths annually 64. Foodssubjected to fecal-oral contamination Undercooked poultry Raw milkNot seen in produce as much due to lack of survival abilities 65. CookingPrevent cross contamination 66. Consumer education 67. 1. Remove pieces of cheese from package. 2. Place in mouth, chew, and swallow. 3. Talk about hurdles and barriers. 68. Topic15 minute exerciseFood microbiology 1 refresherContact APCFoodborne illness 2 overviewpH and Aw testing3 STECs and SalmonellaColiform plating on petrifilms4L. monocytogenes and Campylobacter5 Norovirus and Hepatitis ACheese sampling S. aureus carrier sampling 69. NOROVIRUS CONTENTS 70. 1. Typical Norovirus outbreakFecal, oral, aerosol, and vomitus originsSome form of food contamination occurs Either food is consumed raw or undercooked Person eats the foodIllness starts in 12-48 hrs 71. 1.most common (1-3 days) odiarrhea, vomiting, nausea,stomach pain 2.other symptoms ofever, headache, body aches 72. Non-enveloped, ss-RNA virus Family Calciviridae Infectious dose ~20 viruses 100 billion viruses/g feces in infected person Viruses bind to histological blood proteins (ABO system)(explains below) Humans only reservoir no zoonotic transmission known (yet) Only minor immunity retained, new variations arise frequently 73. Viruses are seeking to invade human cells and reproduce The symptoms are immune reactions to virus presence 74. Cause 80-90% of all virus gastroenteritis USA 5,461,731 est. illnesses annually (food) Approx. 21 million total (food/nonfood) Worldwide 260 million est annually USA 140 deaths annually (mostly elderly) 75. Foods subjected to contaminationleafy greens (such as lettuce), fresh fruits, and shellfish (such as oysters). But, any food that is served raw or handled after being cooked can get contaminated. 76. Hand hygiene Exclusion of ill workers Rapid disinfection 77. Fogger sBleac h 78. 1. 2. 3. 4.Using a sterile moistened swab, swab the interior of your nose. Return swab to 1 ml sample buffer. Vortex to mix well. Place 1 ml swab buffer onto S. aureus Petrifilm. Well incubate films and add test strip as needed, then provide photos of results. 79. Topic15 minute exerciseFood microbiology 1 refresherContact APCFoodborne illness 2 overviewpH and Aw testing3 STECs and SalmonellaColiform plating on petrifilms4L. monocytogenes and Campylobacter5 Norovirus and Hepatitis ACheese sampling S. aureus carrier sampling