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TOXICOLOGY ON FISHERIES PROCESSING– 3 (2 – 1)

DISEASE CAUSED BY MICROORGANISMS. EKO SUSANTO Study Program of Fisheries Processing Technology Diponegoro University Email : eko_thp@undip.ac.id. TOXICOLOGY ON FISHERIES PROCESSING– 3 (2 – 1). EKO SUSANTO – DIPONEGORO UNIVERSITY. Pathogen bacteria that produce toxin. QUESTIONS.

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TOXICOLOGY ON FISHERIES PROCESSING– 3 (2 – 1)

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  1. DISEASE CAUSED BY MICROORGANISMS EKOSUSANTO Study Program of Fisheries Processing Technology Diponegoro University Email : eko_thp@undip.ac.id TOXICOLOGY ON FISHERIES PROCESSING– 3 (2 – 1) EKOSUSANTO – DIPONEGORO UNIVERSITY

  2. Pathogen bacteria that produce toxin

  3. QUESTIONS • Please mention characteristic of C. botulinum? • What happen when butulinum toxin ingested? • Please explain about E. coli based on your knowledge? Please answer those questions for 10 minutes.

  4. CLOSTRIDIUM BOTULINUM Eko Susanto – Diponegoro University EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  5. Gram (+) spore-forming rod • Only srovar A, B, E & F cause botulism in human. • 2 types of C. Botulinum exist :proteolytic (A, some B & F) Non proteolytic (E, some B&) THE ORGANISM EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  6. Gram positive, endospore-forming anaerobes. Botulism is characterized as a rare paralytic disease caused by a nerve toxin produced by the pathogen. The rod-shaped organisms grow best in low-oxygen environments. Proteolityc C. botulinum is a highly dangerous pathogen. Characteristic of C. botulinum

  7. A*= vegetable, fruit, meat, fish & canned products B*= pork meat C = spoil vegetable, carcass, & pork liver D = carcass E*= fish, marine organism, & raw fish F = liver paste Botulinum toxin types EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  8. Botulinogenic properties of fish products (Huss, 1994)

  9. Heat labile proteins. Innactivation at temperature 121oC, freezing doesn’t innactivate botulism toxins. 7 major neurotoxins (types A – G). Neurotoxin are commonly associated with other proteins, such as hemagluttinin & non-toxin-nonhemagglutinin. The neurotoxins are 150-kDa poteins, comprise heavy chain (100 kDa) & light chain (50 kDa). Characteristic of botulinum neurotoxin

  10. Neurotoxin ingested in food Neurotoxin passes through gut mucosa into bloodstream Neurotoxin spreads through body via bloodstream Toxin binds to nerve at the nerve muscle junction Muscle fibre Nerve supply to muscle fiber This block the release of acethylcholine. Muscle cannot to contract, resulting in paralysis WHAT HAPPENS WHEN C. botulinum NEUROTOXIN IS INGESTED ?? (Garbutt, 2007) redesign by Susanto

  11. Mechanism of Botulinum Toxin Botulinum toxin ingested on human

  12. Effect of environmental factors on the growth and survival of proteolytic C. botulinum and non proteolitic C. botulinumb (Peck, 2010) a Heat reistance data without/with lysoyme during recovery b modified from the work of Lund & Peck (2000)

  13. Nausea & vomitting Mainly: neuroligical-burred / blurred vission, difficulty to swallowing (dysphagia), mouth dryness, speech difficulties (dysphonia) & limb & respiration become paralysed, dizziness/vertigo, muscle weakness. Death normally caused by respiratory & cardiac paralysis. Symptoms Botulism EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  14. Botulism toxins are among the most toxic subtances. The minimum lethal dose for mice is 0.4-2.5 ng/kg mouse tissue. 50 % lethal dose for human is 1 ng/g body weight. ex.: 10 people weighing 80 kg each ingested 8.0 x 10-8g of toxin then five of them would beexpected die. Lethal dose of toxin EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  15. 110 100 90 80 70 60 50 40 30 20 10 0 Reported Cases 1982 1987 1992 1997 2002 Year MMWR Center for Food Security and Public Health Iowa State University 2004 EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  16. Recorded food-borne botulism in different countries (peck, 2010) EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  17. EXAMPLE OF RECENT INCIDENTS OF FOOD BORNE BOTULISM INVOLVING NONPROTEOLYTIC c. botulinum (Peck, 2010)

  18. Minimum growth & neurotoxin production occur with range 10oC to 12oC Growth of proteolytic C. Botulinum is prevented at pH of < 4.6 or by 10 % NaCl Minimum aW 0.94 and 0.93 with NaCl & Glycerola. Heat treatment at 121.1oC for 3 min has been adopted as the minimum standard for a botulinum cook CONTROL OF PROTELYTIC c. Botulinum IN FOOD PROCESSING OPERATION EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  19. Minimum growth & neurotoxin production occur with range 3oC to 3.3oC at 5 – 7 weeks. Growth of proteolytic C. Botulinum is prevented at pH of < 5 or by > 5 % NaCl Minimum aW 0.97 and 0.94 with NaCl & Glycerola. Alternative processing technology: high hydrostatic-pressur-treated CONTROL OF nonPROTELYTIC c. Botulinum IN FOOD PROCESSING OPERATION EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  20. Recommended procedures to ensure the safety of minimally heated foods with respect to nonproteolytic C. botulinum EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  21. Stahylococus aureus EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  22. Gram-positive cocci occuring in iregular clumps. Causing intoxication. Optimum growth temperature is 37oC (range of 6 to 48oC). Growth is inhibited in the presence of 0.1% presence acetic acid (pH 5.1) or at pH 4.8 with 5 % NaCl. Able to grow at Aw 0.86. it is capable to grow at Aw 0.83 in the presence of NaCl, sucrose, or glycerol humectants. CHARACTERISTIC OF THE ORGANISM EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  23. Staphylococci are ubiquitous in air, dust, sewage, water, milk, & many foods & on food equipments, environmental surface, human & animals. 30 – 50 % of population are nasal & throat carrier. 15 % are skin carrier (hands esp. patients & staff in hospitals having a carier rate 80%) Skin lesson exp. boils & infection of cuts & burns. Human faeces & clothing. Cows & goat. Food products with high protein is a good growth substrates for S. aureus. Source of MO EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  24. Food-borne outbreaks EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  25. Caused by ingestion of food containing SE performed by metabolically active staphylococci. • Usually a self-limiting illness with short incubation (1-8 h). • The severity depends on : • individual’s susceptibility to the SE, • the amount of contaminated food eaten, • the amount of in the food ingested. • The general health of victim Sfp characteristic EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  26. Nausea Vomitting Diarrhoea & abdominal pain. Collapse & dehydration in severe cases SYMPTOMS EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  27. The minimum amount of toxin required of toxin to produce food poisoning is 1 ng/g (10-9 g/g) of food ingested Lethal dose of toxin EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  28. PREVALENCE OF S. aureus IN SEVERAL FOODS Source: Seo & Bohac, 2010 EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  29. Cooked meats & meat products Foods containing milk of cream Poultry meat & meat products Pre-cooked fish & fish products Pre-cooked crusaceans Gelatine glazes Canned food pasta Food associated with outbreak of S. aureus EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  30. Avoid direct handling of foods (use tongs / gloves) Ensure that raw materials used 4 the prodution of high risk foods are kept refrigerated b4 used. Ensure that high risk foods are rapidly cooled to below 5oC after cooking. Ensure that gigh-risk foods are refrigerate until ready to use. Good personal hygiene. Cover cust / wounds with waterproof dressing Exclude anyone coughing, sneezing / septics cuts / boil from food handling. Prevention of outbreak of S. aureus EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  31. ESCHERICHIA COLI EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  32. Gram negative rod. Member family Enterobacteriaceae family. Able to adapt & colonize a diverse array of environment & the gastrointestinal (GI) E. coli bacteria are mesophilic organism E. coli able to grow at temperature of 10-40oC with optimum tempt 37oC Pathogen can replicate pH values of 4 – 10 & in the presence up t 8% NaCl. Most strains of E.coli are not human pathogen The characteristic of organism EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  33. Environment: • Water sources, compost, urban & rural soils & landscape, sewage, animals include beef & dairy cattle, sheep, swine, horses, rodents, dogs, horses, rodents. • Foods: • Cross contamination to RM, processing water, equipments, & workers. Source of diarrheagenic e. coli EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  34. Entheropathogenic E. coli (EPEC) Enteroinvasive E. coli (EIEC) Enterotoxigenic E. coli (ETEC) Enterohaemorrhagic E. coli (EHEC), also called verocytotoxic E. coli (VTEC) Groups of e. coli Causing gastroenteritis in babies & children Causing Travellers diarrhoea EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  35. Most common mode of transmission, host, symptoms, & characteristics of illness associated with different classes (Beadchamp & Sofos, 2010)

  36. Mechanism of pathogenicity which different e. coli (Beadchamp & Sofos, 2010) EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  37. Diarrhoea & abdominal pain with bleeding – blood appears in stool. Renal failure due to blood cots in the kidney tubules. Internal bleeding due to resulting in brain damage. headache, mild fever, Severe infections of certain serotypes include bloody diarrhea (hemorrhagic colitis), as well as real (kidney) malfunction and failure, thrombocytopenia )inadequate platelet count), microangiopathic hemolytic anemia (lysis of red blood cells), hemolytic uremic syndrome (HUS). Seizure stroke, herniated bowel, chronic rhenal malfuction. symptoms EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  38. Intrinsic factors: • pH, Aw, temperture,nutrients of food. • Extrinsic factors: • Higher level contamination, prevalence in contaminated food products & frequency of catered /picnic/outdoor gilling events. Nationwide distribution of contaminated products causes outbreak Intrinsic & extrinsic factors involved in outbreaks & recalls EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  39. 5 most significant “foodborne illness risk factors” include: • Acquitition of products from unsafe sources • Poor personal hygiene • Contaminated processing equipments • Inadequate heat treatments • Improper handling, processing & storage temperature Food processing condition associated with outbreak EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  40. The infective dose to be low, possibly 10 – 100 organism. E. coli infection strategy Infective dose EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  41. Center for Food Security and Public Health Iowa State University 2004 MMWR EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  42. Advances in diarrheagenic e. coli CONTROL MEASURES DURING PRODUCTION & PROCESSING EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  43. Increased pest control & sanitation programs. Use of HACCP concepts, GAP, GHP, SSOP, GDP. Final products testing for E. coli hide contamination and/or fish meat contamination. SUFFER SUPPLY CHAINS EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  44. Address worker education & hygiene. Zero tolerance policy during processing include equipments, RM, products, & workers. Reduce cross contamination betwen products & equipmentsand/or workers MINIMIZING CONTAMINATION EVENTS DURING PROCESSING EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  45. Appropriate processing & handling Pasteurization Heat treatments Low dose of irradiation (≥ 3 kGy is able to innactivate pathogen). Nonthermal processes: High hydrostatic pressure, shock waves, ultrasonication, pulsed ultraviolet INACTIVATION EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  46. Using antimicrobial agents/ingredients within food products. Using modified atmosphere packaging. Chill temperature (4oC or 40oF)  E. coli not grow INHIBITION EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  47. Do not eat raw / undercooked beef / drink raw milk Abattoir hygiene to prevent faecal contamination adequate cookin of meat & fish Good personal hygiene & health education prevention EKOSUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

  48. Thank you for attention

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