DISEASE CAUSED BY MICROORGANISMS EKO SUSANTO DIPONEGORO UNIVERSITY EKO SUSANTO Study Program of Fisheries Processing Technology Diponegoro University Email ekothpundipacid ID: 913266
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Slide1
TOXICOLOGY ON FISHERIES PROCESSING– 3 (2 – 1)
DISEASE CAUSED BY MICROORGANISMS
EKO SUSANTO – DIPONEGORO UNIVERSITY
EKO
SUSANTOStudy Program of Fisheries Processing TechnologyDiponegoro University Email : eko_thp@undip.ac.id
Slide2Pathogen bacteria that produce toxin
Slide3QUESTIONS
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.
Slide4CLOSTRIDIUM BOTULINUM
4
Eko Susanto – Diponegoro University
EKO SUSANTO –
DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide5THE ORGANISM
Gram (+) spore-forming rodOnly 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&)
EKO
SUSANTO – DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide6Characteristic of C. botulinum
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.
Slide7Botulinum toxin typesA*= vegetable, fruit, meat, fish & canned products
B*= pork meatC = spoil vegetable, carcass, & pork liverD = carcassE*= fish, marine organism, & raw fish
F = liver paste
EKO SUSANTO
– DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide8Botulinogenic properties of fish products (Huss, 1994)
Fish product
Factors adding to botulism hazard
Factors reducing botulism hazard
Safety of product based on
clasification
Fresh and frozen
Vacuum packaging
Traditional chill storage putrefaction before toxin is produced
Cooking before being eaten
No risk
Pasteurized
Prolonged storage life
toxin produced before putrefaction
vacuum packaging
poor hygiene
Chill storage (< 3
o
C)
Synergistic aerobic flora eliminated
Cooking before being eaten
Chill storage
No risk if cooked
High risk if not cooked
Cold smoked
Same as above
Not cooked before being eaten
No tradition for chill storage
Chill storage
Salting (NaCl concentration > 3%)
High redox-potential in unspoiled products
Chill storage
Process control (raw material, salting when applicable)
High risk
Fermented
Fermentation may be slow
High temperature during fermentation
Not cooked before being eaten
Salting (NaCl concentration 3 % in brine)
Chill storage
Low pH
Process control
Chill storage
High risk
Semi-preserved
Not cooked before being eaten
Application of salt, acid etc
Chill storage
Process control
Low risk
Fully preserved
Not cooked before being eaten
Packed in closed cans
Autoclaving
Process control (autoclaving, closing of cans)
Low risk
Slide9Characteristic of botulinum neurotoxinHeat 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).
Slide10WHAT HAPPENS WHEN C. botulinum NEUROTOXIN IS INGESTED ??
Nerve supply to muscle fiber
Neurotoxin spreads through body via bloodstream
Toxin binds to nerve at the nerve muscle junction
Muscle fibre
This block the release of acethylcholine. Muscle cannot to contract, resulting in paralysis
Neurotoxin passes through gut mucosa into bloodstream
Neurotoxin ingested in food
(Garbutt, 2007)
redesign by Susanto
Slide11Mechanism of Botulinum ToxinBotulinum toxin ingested on human
Slide12Effect of environmental factors on the growth and survival of proteolytic C.
botulinum and non proteolitic C. botulinum
b (Peck, 2010)
Factor
Proteolytic C. botulinum
Nonproteolytic C. botulinum
Neurotoxin formed
A,B,F
B,E,F
Minimum growth temperature
10-12
o
C
2.5-3.0
o
C
Maximum growth temperature
37
o
C
25
o
C
Minimum pH for growth
4.6
5.0
NaCl concentration preventing growth (%)
10
5
Minimum water activity for growth
NaCl as humectant
0.94
0.97
Glycerol as humectant
0.93
0.94
Spore heat resistant
D
121
o
C
= 0.21 min
D
82.2
o
C
= 2.4/231 min
a
Spore radiation resistant
D = 2.0 – 4.5 kGy
D = 1.0 – 2.0 kGy
Food involved in botulism outbreak
Home-canned foods, faulty commercial processing
Fermented marine products, dried fish, vacuum-packed fish
a
Heat reistance data without/with lysoyme during recovery
b
modified from the work of Lund & Peck (2000)
Slide13Symptoms Botulism
Nausea & vomittingMainly: 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.
EKO SUSANTO
– DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide14Lethal dose of toxinBotulism 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.
EKO
SUSANTO – DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide15Center for Food Security and Public Health Iowa State University 2004
Year
1982 1987 1992 1997 2002
Reported Cases
1101009080706050403020
100
MMWR
EKO
SUSANTO
–
DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide16Recorded food-borne botulism in different countries (peck, 2010)
Country
Period
No. of cases
Total
Avg per yr
Belgium
1982 – 2000
32
2
Canada
1971 – 2005
439
13
China
1958 – 1983
4377
168
Denmark
1984 – 2000
18
1
France
1971 – 2003
1286
39
Georgia
1980 – 2002
879
40
Germany
1983 – 2000
376
22
Italy
1979 – 2000
750
34
Japan
1951 – 1987
479
13
Poland
1971 – 2000
9219
307
Spain
1971 – 1998
27
10
Sweden
1969 – 2000
13
1
United Kingdom
1971 – 2005
38
1
United States
1971 – 2003
934
28
EKO
SUSANTO
–
DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide17EXAMPLE OF RECENT INCIDENTS OF FOOD BORNE BOTULISM INVOLVING NONPROTEOLYTIC c.
botulinum
Yr
Country
Product
Toxin type
No. of cases
Factor contributing to botulism outbreak
References
1991
Egypt
Commercial unviscerated, salte fish
E
> 81
Putrefaction of fish before salting
Weber et al (1993)
1997
Fance
Fish
E
1
Not known
Korkeala et al., 1998
1997
Germany
Home-smokked, vacuum packed fish
E
4
Temperature abuse
Anonymous (1998b)
1998
France
Commercial frozen, vacum packed scallop
E
1
Temperature abuse
Boyer et al (2001)
2001
Canada
Hommade fermented samon roe
E
4
Unsafe process
Anoymous 2002
2003
Germany
Home-salted, air-dried fish
E
3
Temperature abuse
Eriksen et al (2004)
2004
Germany
Commercial vacuum packed smoked salmon
E
1
Consumed after “use by date”
Dressler (2005)
(Peck, 2010)
Slide18CONTROL OF PROTELYTIC c. Botulinum
IN FOOD PROCESSING OPERATION
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 % NaClMinimum 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
EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide19CONTROL OF nonPROTELYTIC c. Botulinum
IN FOOD PROCESSING OPERATION
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 % NaClMinimum aW 0.97 and 0.94 with NaCl & Glycerola.Alternative processing technology: high hydrostatic-pressur-treated
EKO
SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide20Recommended procedures to ensure the safety of minimally heated foods with respect to nonproteolytic
C. botulinum
Recommendation
Storage at < 3.0
oCStorage at ≤ 8 oC and shelflife of ≤ 10
oC
Storage at chill temperature combined with heat treatment of 90
oC for 10 min or equivalent lethality (e.g., 80
oC for 129 min, 85o
C for 36 min
Storage of chill temperature combined with pH ≤ 5.0 throughtout the food
Storage of chill temperature combined with a salt concentration of ≥ 3.5% throughtout the food
Storage at chill temperature combined with a combination of heat treatment and other preservative factors which can be shown consistenly to prevent growth and neurotoxin production by
C. botulinum
EKO
SUSANTO
–
DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide21Stahylococus aureus
EKO
SUSANTO – DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide22CHARACTERISTIC OF THE ORGANISM
Gram-positive cocci occuring in iregular clumps.Causing intoxication.Optimum growth temperature is 37oC (range of 6 to 48o
C).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.
EKO
SUSANTO
– DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide23Source of MOStaphylococci 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.
EKO
SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide24Food-borne outbreaks
EKO
SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide25Sfp characteristicCaused 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
EKO SUSANTO
– DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide26SYMPTOMSNauseaVomitting
Diarrhoea & abdominal pain.Collapse & dehydration in severe cases
EKO SUSANTO
– DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide27Lethal dose of toxinThe minimum amount of toxin required of toxin to produce food poisoning is 1 ng/g (10
-9 g/g) of food ingested
EKO SUSANTO –
DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide28PREVALENCE OF S. aureus IN SEVERAL FOODS
Products
No. of samples tested
% positive for S. aureus
No. of S. aureus CFU/g
% Positive for SE
Reference
Raw meat
139
2.8
NA
7.8
Moon et al. (2007)
Raw milk
714
7.9
NA
31.8
Moon et al. (2007)
Frozen prawn
46
23.9
>3
NA
Sanjeev et al. (1987)
Shrimp
1,468
27
>3
NA
Swartzentruber et al. (1980)
Ready-to-eat fast food
3,332
8.6
NA
47
Oh et al. (2007)
Source: Seo & Bohac, 2010
EKO
SUSANTO
–
DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide29Food associated with outbreak of S. aureus
Cooked meats & meat productsFoods containing milk of cream
Poultry meat & meat productsPre-cooked fish & fish productsPre-cooked crusaceansGelatine glazesCanned foodpasta
EKO
SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide30Prevention of outbreak of S. aureus
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 dressingExclude anyone coughing, sneezing / septics cuts / boil from food handling.
EKO
SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide31ESCHERICHIA COLI
EKO
SUSANTO – DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide32The characteristic of organism
Gram negative rod.Member family Enterobacteriaceae family.Able to adapt & colonize a diverse array of environment & the gastrointestinal (GI)E. coli
bacteria are mesophilic organismE. coli able to grow at temperature of 10-40oC with optimum tempt 37oCPathogen can replicate pH values of 4 – 10 & in the presence up t 8% NaCl.Most strains of E.coli are not human pathogen
EKO
SUSANTO
– DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide33Source of diarrheagenic e. coli
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.
EKO SUSANTO
– DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide34Groups of e. coli
Entheropathogenic E. coli (EPEC)Enteroinvasive
E. coli (EIEC)Enterotoxigenic E. coli (ETEC)Enterohaemorrhagic E. coli (EHEC), also called verocytotoxic E. coli (VTEC)
Causing gastroenteritis
in babies & childrenCausing Travellers diarrhoea
EKO
SUSANTO – DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide35Most common mode of transmission, host, symptoms, & characteristics of illness associated with different classes
Class
Classic host
Symptoms
Incubation duration (days)
Acute / chronic presentation
Infection dose
EPEC
Infants
(< 6 mo); more prevalent in deveeloping countries
Severe diarrhea, fever, vomiting
Variable
Chronic diarrhea, malnutrition
High; low in fant
EAEC
Children; moe prevalent in devloping countries
Watery or bloody diarrhea, fever
Variable
Chronic watery diarrhea, severe dehydration
High
EIEC
Children, more prevalent in developing countries
Watery diarrhea, abdominal cramping, fever
1-3; self limiting
Dysentry syndrome
Low
ETEC
Traveers & infants native to developing countries
Watery diarrhea, abdominal cramping, milk fever, nausea
1-3; 3-7
Cholera – like
High
EHEC
C
hildren & elderly
Diarrhea, bloody diarhea, abdominal pain, vomiting
1-8; 4-10
Bloody diarrhea (hemorrhagic colitis), HUS, kidney failure
low
(Beadchamp & Sofos, 2010)
Slide36Mechanism of pathogenicity which different e. coli
Class
adhesion site
Adhesion mediator
Invassion potential
Toxins
Other virulence factor
EPEC
Small intestine
Intimin
Moderate
Possible enterotoxin (EAST1)
EAF palsmid, LEE island, flagellin, CDT
EAEC
Large & small intestine
AAF
None
EAST1, Pet, Pic
Flagellin
EIEC
Large intestine (colon)
Unclear
High
Enterotoxin
Cell-to-cell spread )IcsA), serine-protease (SepA)
ETEC
Small intestine
Fibrial CFs
None
LT, ST
CDT
EHEC
Large intestine (colon)
Intimin
Moderate
Stx, enterohemolysin, EAST1
LEE island, pO157, flagellin, CDT, CNF
(Beadchamp & Sofos, 2010)
EKO
SUSANTO
–
DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide37symptoms
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.
EKO
SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide38Intrinsic & extrinsic factors involved in outbreaks & recalls
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
EKO
SUSANTO –
DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide39Food processing condition associated with outbreak5 most significant “foodborne illness risk factors” include:
Acquitition of products from unsafe sourcesPoor personal hygieneContaminated processing equipmentsInadequate heat treatments
Improper handling, processing & storage temperature
EKO SUSANTO
– DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide40Infective dose The infective dose to be low, possibly 10 – 100 organism.
E. coli infection strategy
EKO SUSANTO
– DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide41Center for Food Security and Public Health Iowa State University 2004
MMWR
EKO
SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide42Advances in diarrheagenic e. coli
CONTROL MEASURES DURING PRODUCTION & PROCESSING
EKO
SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide43SUFFER SUPPLY CHAINSIncreased 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.
EKO
SUSANTO – DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide44MINIMIZING CONTAMINATION EVENTS DURING PROCESSINGAddress worker education & hygiene.
Zero tolerance policy during processing include equipments, RM, products, & workers.Reduce cross contamination betwen products & equipmentsand/or workers
EKO
SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide45INACTIVATIONAppropriate processing & handling
PasteurizationHeat treatmentsLow dose of irradiation (≥ 3 kGy is able to innactivate pathogen).Nonthermal processes: High hydrostatic pressure, shock waves, ultrasonication, pulsed ultraviolet
EKO
SUSANTO – DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide46INHIBITIONUsing antimicrobial agents/ingredients within food products.
Using modified atmosphere packaging.Chill temperature (4oC or 40oF)
E. coli not grow
EKO
SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)
Slide47prevention
Do not eat raw / undercooked beef / drink raw milkAbattoir hygiene to prevent faecal contamination adequate cookin of meat & fish Good personal hygiene & health education
EKO
SUSANTO – DIPONEGORO
UNIVERSITY (eko_thp@undip.ac.id)
Slide48Thank you for attention