Food microbiology workshop UEHA 2013

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This is a large presentation for an Environmental Health Specialists workshop held in Utah in 2013. It includes video attachments, quizzes and more.

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  • CDC data from 2011 indicate that there are approximately 5,461,731 cases of Norovirus per year. [read slide].
  • The 2011 data from the CDC indicates that there are approximately 149 deaths attributed to Norovirus annually. While this may seem high compared to the others on this table, it is out of over 5 million illnesses. Most likely those that died were highly immuno-compromised and Norovirus contributed to their death rather than caused their death. Salmonella is considered the most deadly at 378 deaths per year. Toxoplamagondiia unicellular parasite is implicated in 327 deaths annually. Listeria monocytogenes causes 255 deaths per year and this is out of only approximately 1500 illnesses. This is the pathogen with the highest percentage of death. Clostridium perfringens causes 26 deaths per year. Campylobacter causes 76 deaths per year. The Shiga toxin producing E. coli like E. coli O157:H7 only cause 20 deaths per year. Shigella causes 10 deaths per year. Yersinia enterolcoliticacauses 29 deaths per year. The combined group of Vibrios cause only 48 deaths per year. No deaths were attributed to Trichinella illness. And lastly, one of the deadly toxin producers, Clostridium botulinum only causes 9 deaths annually.
  • There are over 250 different food pathogens identified to date. However, most are rarely encountered and little data is available on them. There are 31 more highly known foodborne pathogens. Most of these pathogens are tracked by local, state, and federal public health officials. Data in 2006 shows that Norovirus causes approximately 40% of all foodborne illnesses. The remaining four pathogenic viruses are not tracked in this figure. Bacteria including 21 different bacterial pathogens makes up 23% of all foodborne illnesses. Five different parasite pathogens account for just 1% of foodborne illnesses. Chemicals cause approximately 5% of foodborne illnesses. At least 29% of foodborne gastroenteritis illnesses have no cause. This may be due to insufficient data or unidentified pathogens. There is no data on potential foodborne pathogens causing non-gastroenteritis symptoms.
  • The remainder of the course will take an in-depth look at these fourteen different foodborne illness pathogens or pathogen groups.
  • A typical outbreak of Staph gastroenteritis starts with a food that is cooked, for example ham. This will kill off any competing microorganisms. Then a person handling the ham, maybe making ham salad, contaminates it with their hands or with a sneeze. The ham salad is then left out at room temperature. Maybe it will be taken to a picnic and sit in the sun for a few more hours. Finally the picnickers dine on the ham salad. They enjoy their outing and return home. In just a hour or more after eating people will know something is wrong. Something is very wrong. Hopefully they are not in their car on an hour long ride home.
  • A typical outbreak of Staph gastroenteritis starts with a food that is cooked, for example ham. This will kill off any competing microorganisms. Then a person handling the ham, maybe making ham salad, contaminates it with their hands or with a sneeze. The ham salad is then left out at room temperature. Maybe it will be taken to a picnic and sit in the sun for a few more hours. Finally the picnickers dine on the ham salad. They enjoy their outing and return home. In just a hour or more after eating people will know something is wrong. Something is very wrong. Hopefully they are not in their car on an hour long ride home.
  • Food microbiology workshop UEHA 2013

    1. 1.   UEHA 2013 Special Workshop
    2. 2. Topic 15 minute exercise Food microbiology 1 refresher Contact APC Foodborne illness 2 overview pH and Aw testing 3 STECs and Salmonella Coliform plating on petrifilms 4 L. monocytogenes and Campylobacter 5 Norovirus and Hepatitis A Cheese sampling S. aureus carrier sampling
    3. 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. We’ll collect these plates and incubate them, then provide pictures of the results.
    4. 4. Topic 15 minute exercise Food microbiology 1 refresher Contact APC Foodborne illness 2 overview pH and Aw testing 3 STECs and Salmonella Coliform plating on petrifilms 4 L. monocytogenes and Campylobacter 5 Norovirus and Hepatitis A Cheese sampling S. aureus carrier sampling
    5. 5. 1 in 3 1 in 5 1 in 6 1 in 12
    6. 6. Annual foodborne illnesses 2011 Norovirus 5,461,731 Yersinia enterocolitica Salmonella spp 1,027,561 Unicellular parasites Clostridium perfringens 965,958 Vibrios Campylobacter 845,024 Listeria monocytogenes Staphylococcus aureus 241,148 Trichinella STEC 175,905 C. botulinum Shigella 131,254 97,752 ~70,000 52,408 1591 156 55
    7. 7. 50 600 3,000 40,000
    8. 8. Annual foodborne illness deaths Norovirus Salmonella spp Clostridium perfringens Campylobacter Staphylococcus aureus STEC Shigella 149 Yersinia enterocolitica 378 Unicellular parasites 26 Vibrios 76 Listeria monocytogenes 6 Trichinella 20 C. botulinum 10 29 327 48 255 0 9
    9. 9. Salmonellosis Botulism Cholera / Typhoid Fever Norovirus Gastroenteritis
    10. 10. Salmonellosis Botulism Cholera / Typhoid Fever Norovirus Gastroenteritis
    11. 11. Meats Poultry Eggs All answers are correct
    12. 12. 33 25 >250 2,033
    13. 13. 4% 40% 14% 44%
    14. 14. 2% 12% 18% 21%
    15. 15. Other/Multiple 2% Unknown 29% Norovirus 40% Chemical 5% Parasites 1% Bacteria 23%
    16. 16. True False
    17. 17. Listeria monocytogenes Norovirus Salmonella Clostridium botulinum
    18. 18. Very young Very old Infirm (sick) Allergic individuals
    19. 19. Stomach acids Immune System Intestinal flora competition Just pure luck
    20. 20. Where a pathogens’ toxin causes illness Where 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. 21. Global food sources Consumer choice for fresh foods Emerging pathogens Lack of on-farm food safety
    22. 22. Foodborne illnesses Norovirus 1 Yersinia enterocolitica 8 Salmonella spp 2 Unicellular parasites 9 Clostridium perfringens 3 Vibrios 10 Campylobacter 4 Listeria monocytogenes 11 Staphylococcus aureus 5 Trichinella 12 STEC 6 C. botulinum 13 Shigella 7 Non-infectious 14
    23. 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. We’ll collect these plates and incubate them, then provide pictures of the results.
    24. 24. Topic 15 minute exercise Food microbiology 1 refresher Contact APC Foodborne illness 2 overview pH and Aw testing 3 STECs and Salmonella Coliform plating on petrifilms 4 L. monocytogenes and Campylobacter 5 Norovirus and Hepatitis A Cheese sampling S. aureus carrier sampling
    25. 25. 1. Remove 5 ml of milk from sample container and pipet onto coliform petrifilm. 2. Press petrifilm with plastic tool. 3. We’ll collect these films, incubate them, then provide pictures of the results.
    26. 26. E. Coli O157: H7 CONTENTS
    27. 27. 1. ETEC – Enterotoxigenic -Travellers diarrhea a. 2. 3. 4. Has toxins that cause diarrhea EPEC – 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. 28. 1. Typical STEC EHEC outbreak Fecal origins of E. coli Some form of cross contamination occurs Either food is consumed raw or undercooked Person eats the food Illness starts in 3-4 days, but can be 1-9 days
    29. 29. • Mild • • Hemorrhagic colitis (2-9 days) • • Asymptomatic to mild diarrhea Abdominal cramps and dysentery Hemolytic 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. 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. 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. 32. • • • USA 175,905 est. STEC illnesses annually Approx 63,000 are EHEC 20 deaths annually
    33. 33.  Foods • subjected to fecal-oral contamination Undercooked 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. 34. Cooking Prevent cross contamination
    35. 35. Consumer education
    36. 36. CONTENTS
    37. 37. 1. Typical outbreak Salmonella is a classic fecal contaminate Some form of cross contamination occurs Processing fails to kill all Salmonella Person eats the food Illness starts in 6-72 hours
    38. 38. • Common • Diarrhea & abdominal cramps • Fever • Additional symptoms: • • • • • Bloody diarrhea Vomiting Headache Body aches Reactive arthritis
    39. 39. • • • • G-, non-sporeformer, facultative, rod Survives difficult environments (dry and acid) Predominantly fecal S. enterica Enteritidis and S. enterica Typhimurium • • >2500 serotypes Requires large numbers for infection OR only 1 if protected from stomach acids
    40. 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 – don’t see high levels of DNA transfer
    41. 41. • USA 1,027,561 591 est. illnesses/yr • Second largest number after Norovirus • 378 deaths annually • Deaths in immunocompromised • • Use PFGE to quickly determine serovar and strain Used to implicate multistate outbreaks
    42. 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. 43. Proper cooking Prevent cross contamination
    44. 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. 45. Consumer education
    46. 46. Topic 15 minute exercise Food microbiology 1 refresher Contact APC Foodborne illness 2 overview pH and Aw testing 3 STECs and Salmonella Coliform plating on petrifilms 4 L. monocytogenes and Campylobacter 5 Norovirus and Hepatitis A Cheese sampling S. aureus carrier sampling
    47. 47. CONTENTS
    48. 48. 1. Typical outbreak Listeria monocytogenes found in food processing environment Some form of cross contamination occurs Lm can grow slowly in foods even under refrigeration Person eats the food Illness starts in >12 hours, but can be several days
    49. 49. • Gastrointestinal • Mild flu-like symptoms (persistent fever, mild nausea, mild vomiting) • Invasive (enters phagocytic immune cells) • Septicemia • Meningitis • Spontaneous abortion barrier) (crosses placental
    50. 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. 51. • Many different serovars • one highly virulent (4b) • while others not at all • • Requires ↑ no.s for infection due to success of human immune system Virulence associated with pathogenic factors (genes
    52. 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. 53. • • • USA 1591 est. illnesses annually 255 deaths annually Highest FBI death rate (~16%)
    54. 54.  Foods that 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. 55. Sanitation Prevent cross contamination
    56. 56. Anti-listerial additives Post packaging thermal processing
    57. 57. Consumer education
    58. 58. Campylobacter CONTENTS
    59. 59. 1. Typical outbreak Fecal origins in poultry, swine, sheep and cattle Some form of cross contamination occurs Either food is consumed raw or undercooked Person eats the food Illness starts in 2-5 days
    60. 60. • Gastrointestinal (2-10 days) • • • Bacteremia (0.15% progress) • • Fever, diarrhea, cramps, vomiting May have muscle aches, nausea, headache Meningitis, hepatitis, pancreatitis Guillan 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. 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. 62. • • • Adhesion proteins Invasion proteins Toxin proteins • CDT cytolethal distending toxin • Hemolysin toxin
    63. 63. • • • • USA 845,024 est. illnesses annually Most undiagnosed Most in very young 1-12 months and other immunocompromised 76 deaths annually
    64. 64. Foods subjected to fecal-oral contamination • • • Undercooked poultry Raw milk Not seen in produce as much due to lack of survival abilities
    65. 65. Cooking Prevent cross contamination
    66. 66. Consumer education
    67. 67. 1. Remove pieces of cheese from package. 2. Place in mouth, chew, and swallow. 3. Talk about hurdles and barriers.
    68. 68. Topic 15 minute exercise Food microbiology 1 refresher Contact APC Foodborne illness 2 overview pH and Aw testing 3 STECs and Salmonella Coliform plating on petrifilms 4 L. monocytogenes and Campylobacter 5 Norovirus and Hepatitis A Cheese sampling S. aureus carrier sampling
    69. 69. NOROVIRUS CONTENTS
    70. 70. 1. Typical Norovirus outbreak Fecal, oral, aerosol, and vomitus origins Some form of food contamination occurs Either food is consumed raw or undercooked Person eats the food Illness starts in 12-48 hrs
    71. 71. 1. most common (1-3 days) odiarrhea, vomiting, nausea, stomach pain 2. other symptoms ofever, headache, body aches
    72. 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. 73. • • Viruses are seeking to invade human cells and reproduce The symptoms are immune reactions to virus presence
    74. 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. 75.  Foods • • • • subjected to contamination leafy 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. 76. • • • Hand hygiene Exclusion of ill workers Rapid disinfection
    77. 77. Fogger s Bleac h
    78. 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. We’ll incubate films and add test strip as needed, then provide photos of results.
    79. 79. Topic 15 minute exercise Food microbiology 1 refresher Contact APC Foodborne illness 2 overview pH and Aw testing 3 STECs and Salmonella Coliform plating on petrifilms 4 L. monocytogenes and Campylobacter 5 Norovirus and Hepatitis A Cheese sampling S. aureus carrier sampling

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