toxicology on fisheries processing– 3 (2 – 1)

TOXICOLOGY ON FISHERIES PROCESSING– 3 (2 – 1)

DISEASE CAUSED BY MICROORGANISMS (1st meeting)

EKO SUSANTO – DIPONEGORO UNIVERSITY

EKO SUSANTOStudy Program of Fisheries Processing Technology

Diponegoro University Email : eko_thp@undip.ac.id

REFERENCES: Peck, M.W., 2010. Clostridium botulinum. Edited by: Juneja,

K.V., and Sofos, K.N. Pathogens and Toxin in Food: challenges and intervention. ASM Press. Washinton DC.

Juneja, K.V., Novak, J.S., and Labbe, R.J, 2010. Clostridium perfringens. Edited by: Juneja, K.V., and Sofos, K.N. Pathogens and Toxin in Food: challenges and intervention. ASM Press. Washinton DC.

Beauchamp, C.S. and Sofos, J.N. 2010. Diarahegenic Eschericia coli. Edited by: Juneja, K.V., and Sofos, K.N. Pathogens and Toxin in Food: challenges and intervention. ASM Press. Washinton DC.

Seo, K.S. Bohach, G.H., 2010. Staphylococal Food Poisoning. Edited by: Juneja, K.V., and Sofos, K.N. Pathogens and Toxin in Food: challenges and intervention. ASM Press. Washinton DC.

Wright, A.C. and Sceneider, K.R. 2010. Pathogenic vibrios in seafood. Edited by: Juneja, K.V., and Sofos, K.N. Pathogens and Toxin in Food: challenges and intervention. ASM Press. Washinton DC.

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

REFERENCES: CONTINUE Amastrong, G.D. 2008. Pathogenic Mechanisms of the

Enterohemorrhagic Escherichia coli—Some New Insights. Edited by: Wilson, C.L. Microbial Food Contamination 2nd ed. CRC Press. Boca Raton.

Nilsson, L. and Gram, L. . 2002. Improving the control of pathogens in fish products. Edited by: Bremmer, A.H. Safety and quality issues in fish processing. CRC Press. Boca Raton.

WHO-FAO. 2005. Microbiological risk assessment series: Risk assessment of Vibrio vulnificus in raw oysters interpretative summary and technical report. WHO-FAO UN.

Garbutt, J. 1997. Essentials of food microbiology. Arnold. London.

Pelczar, M.J. & Chan, E.C.S. 1976. Dasar-dasar mikrobiologi. Diterjemhakan: Hadioetomo et al., 1988. UI press.

Huss, H.H. 1994. Assurance of seafood quality. FAO fisheries technical paper.

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

LECTURE RULES The lecture will be taken place during 4 meetings 10 minutes after lecturer starting lecture. Students

are prohibited to get in class. If the lecturer is late 10 minutes after the start of

lecture time without confirmation to students, the students are permitted to leaving class.

Final score consist of 35 % tasks and 65 % of final examination

The students have to attend lecture 75 % minimally. The lecture consist of class lecture and self study. The students are permit to get out class during

lecture if they don’t want to joining lecture. Eko Susanto – Diponegoro University

4EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

MATERIALS 1st meeting : bacteria toxins in food. 2nd meeting : Clostridium botulinum toxin

toxin, Staphylococcus aureus toxin, Eschericia coli toxin.

3rd meeting : Vibrio toxin, Pseudomonas cocovenenans toxin, Fungal toxin (Penicillium).

4th meeting : Presentation

5EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

QUESTIONS How do we know if food is being

contaminated by bacteria? Please mention bacteria which cause

food borne? What are the differences between

endotoxin and exotoxin?

Please answer those questions for 10 minutes.

Kerjakan selama 10 menit6EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

INTRODUCTION Food-borne diseases are of major concern to consumers, producers

and authorities alike. Despite an increased awareness, the number of cases and outbreaks

does not appear to be decreasing. Many foods are implicated in food-borne disease outbreaks. Seafoods rank third on the list of products which have caused food-

borne disease. Seafoodborne disease may be caused by a variety of agents, including

aquatic toxins, biogenic amines, bacteria, virus and parasites. Bacteria are mostly found in low numbers in live fish with the exclude

of marine vibrios. Marine vibrios, such as V. parahaemolyticus and V. vulnificus, may be

found in high numbers in shellfish and in shellfish-eating fish from tropical waters and during the summer months in temperate zones

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

DEFINITION

Disease any harmful change in the tissues and/or metabolism of a plant, animal & human that produces the symptoms of illness.

Pathogens MO that cause disease. Toxin chemical substances produced by MO

that are harmful to human tissues and physiology.

Food poisoning an acute (arising suddenly and of short duration) gastroenteritis caused by the ingestion of food (source: Garbutt, 1997

THE NATURE OF FOOD SPOILAGE

Major reasons 4 food being rejected: Organoleptic changes growth MO Chemical changes in food Physical damage Freezer burn ‘staling’ changes Aw Ripening Presence of foreign materials Contamination with chemical agent

FACTORS THAT CAUSE BACTERIA GROWTH Internal factor (Aw, pH, redox potential,

nutrition, natural antibacteria, biology structure) Processing factors (hot treatment, irradiation,

pasteurization, grinding) External factors (environment, temperature) Implicit factor (compt become dominant) Factors of food

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

INTERACTIONS INVOLVED IN THE SELECTION OF SPOILAGE MICROFLORA

Extrinsic factor Contaminating microflora

Implicit factors Intrinsic factors

Growth of specific spoilage microflora

Spoilage symptoms

Interaction

Source: Garbutt, 1997

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

MO CHANGES ON STORED FOOD

0 1 2 3 4 5 6 7 8 9 100

20

40

60

80

100

Bakteri gram (-) Bakteri gram (+)

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

MO CHANGES ON STORED FOOD ON COLD TEMPERATURE

0 1 2 3 4 5 6 7 80

20

40

60

80

100

Bakteri gram (-) Bakteri gram (+)

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

SOURCES OF CONTAMINATION OF FOOD

Raw material with natural micro flora

AirSpoiled foods

Processing equipment

Diseased plants & animals

Packaging materials

Pests Humans

Sewage

Soil

Water

Feces

Aerosol

Dust

Source: Garbutt, 1997

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

IF WE TAKE FRESH FISH FILLETS STORED ON ICE. WHAT ARE THE POTENTIAL SOURCES OF CONTAMINATION?

Natural surface & gut flora of fish Water & possibly sediment from natural habitat Fishing nets Surfaces on board the fishing vessels Fish boxes Ice / refrigerated sea water Human resources Pests Soil air

COMPOSITION OF CONTAMINATING MO Gram-negative rods & coccobacilli.

Acinetobacter, aeromonas, Alcaligenes, Citrobacter, Enterobacter, Escherichia, Flavobacterium, Moraxella, Proteus, Pseudomonas, Salmonella, Shewanella, & Yersinia

Gram positive rodsBacillus, Brochothrix, Clostridium, Corynebacterium, Lactobacillus, & Listeria

Gram negative cocciEnterococcus, Lactococcus, Pediococcus, &

Staphylococcus.

PATHOGENESIS OF FOODBORNE & RELATED ORGANISM

Skeletal muscle : Trichinella spiralis Stomach : Helicobacter pylori Liver : Clonorchis Small intestine :

Astroviruses, Bacillus cereus, Campylobacter jejuni, Clostridium perfringens, E. coli, Salmonellae, S.typhi, Vibrio cholerae, V.parahaemolyticus

Large intestine/colon : Campylobacter (small intestine), E.coli, Entamoeba histoytica, Salmonella eneritidis, Shigellae, especially S. dysenteridae.

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

CHANGES IN FOODS CAUSED BY SPOILAGE MO

General appearance moldy (F) & slimy (B) Color F (red/black) & B (colored); chemical

changes --> greening of meat (H2S) Texture Pseudomonas fluorescens at fish -->

prod proteinase caused tissues to soften Odor / flavor MO prod chem. associated with

metab. act. A mixture of the above

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

HOW DO MO CAUSE DISEASE?1. The permanent Mo is essential in combating

invasion of the body by potential pathogens by competing 4 space & nutrients, stms producing antibiotics. Ex: E.coli prevent salmonellae in the colon

2. Bacteria in the colon synthesize vit. K & contribute significantly to our req. of protein

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

DISEASE PRODUCTION BY MO

Exotoxin: toxin is secreted

by MO into the cell environment

Endotoxin: toxin is produced

by MO & secreted if the MO cell being damaged.

Sifat Exotoxin Endotoxin

MOgram (+) & gram (-) gram (-)

Source protein lipopolisakarida

ketahanan panas

Inactive at 60 - 80 oC, exclude sev exotoxin

stabile at sterilization temp

Lethal doserendah, sangat toksik

> tinggi eksotoksin

cara kerjaSpesifik untuk sel/tenunan sel < spesifik

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

EXOTOXIN

Characteristic: 1. Generally proteins

synthesized by metabolic activity.

2. Produced by gram-positive & gram-negative organism.

3. No structural components of the cell

4. Secreted into the cell environment.

Exotoxin

Exotoxin – soluble protein released into environment

by active cell

Enterotoxin Neurotoxin

Affects the gut Affects the nervous system

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

Source: Garbut, 1997

ENDOTOXIN

Characteristic: 1. Lypopolysacharid

es.2. Toxic components

of the cell wall released when the cell dies & breaks down.

3. Produced by gram-negative MO

Exotoxin

Act as an enterotoxin in the gut

Fever Toxic shock

Rash Inflammation of organ

Endotoxin

Lipopolysaccharide in outer wall layer

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

Source: Garbut, 1997

FOOD POISONING Food poisoning an acute (arising suddenly and of

short duration) gastroenteritis caused by the ingestion of food (Garbutt, 2007).

Gastroenteritis is characterized by: Abdominal pain; Diarrhoea With / without vomiting With / without fever

Bacteria caused food poisoning: S. aureus, C. perfringens, C.botulinum, & Bacillus cereus.

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

INTOXICATION & INFECTION

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

INTOXICATION

Intoxications involve food poisoning in which the organism grows in the food & releases a toxin from cells.Toxin is ingested along with the food, toxin gives rise to the food poisoning syndrome.Bacteria toxins that produce intoxication are exotoxins. S aureus & C botulinum

Active organism secretes exotoxin (enterotoxin ) into food

Food eaten

Enterotoxin affect gut giving gastroenteritis

Enterotoxin affect gut giving gastroenteritis

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

Source: Garbut, 1997

INFECTIONS

Infection involve food poisoning caused by ingestion of live organism when the organisms grow in the gastrointestinal tract to produce the disease. Most food poisoning caused by infection. Ex. Salmonella spp & C. perfringens

Organism ingested along with food

Dose sufficient to overcome host defenses

Organism grows in the host gut

Organism affects gut giving gastroenteritis

Organism appears in faeces in large number

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

PHYSIOLOGICAL MECHANISM ASSOCIATED WITH FOOD POISONING (GARBUTT, 1997)

NORMAL GUT PHYSIOLOGY ENTEROTOXIN INGESTED WITH FOOD

Vomit receptor

Connective tissue

H2O

Na+

Gut epitheliumBlood supply

Enterotoxin affects vomit receptors

H2O out

Na+ out

Vomiting centre in the brain stimulated

Vomiting

Diarrhoea

Fluid and electrolyte loss

Dehydration

Ingestion can lead to people on the situation: Acute illness with med treatment (MT) Acute illness without MT Mild illness without MT, symptoms can be

ignored Infection without symptom

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

FACTORS CAUSE PERSON BECOME ILL

Age Diet Nutrition’ Genetic make-up of the person The presence of other disease Suppressed immunity Previous contact with the disease

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

MAJOR FOOD POISONING ORGANISMS ASSOCIATED WITH SEAFOOD ORGANISM CAUSING DISEASE (NILSSON & GRAM,

2002)Organism causing disease MID of toxin or live cells Primary habitat

Bacteria of aquatic originClostridium botulinum type E 0.1–1 μg toxin Ubiquitous in aquatic

environment, soil, ocean

sediment, intestinal tract of fish,

surface of fish Marine Vibrio sppV. cholera 108 cfu/g Estuarine and coastal warm

waters (>15ºC), intestines of

shellfish-eating fish and tract of

oysters

V. parahaemolyticus 105–106 cfu/gV. vulnificus Unknown

Histamine producing bacteria >100 mg histamine/100 g Members of Enterobacteriaceae from the aquatic environment

Dinoflagellates; maybe bacteria associated with the algae

Paralytic shellfish poisoning (PSP) toxin

Aquatic environment, accumulated in bivalves (e.g. mussels, oysters)

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

CONTINUED:Bacteria from the general environment Listeria monocytogenes unknown–108 cfu/g Widespread in nature, soil,

foilage, faeces, seafood processing environments

C .botulinum (mesophilic) Widespread in soilBacteria from the human/animal reservoirShigella spp. 102–105 cfu/g Faecal polluted coastal regions or

ponds; cause faecal contamination of seafood

Salmonella spp. 10–106

Escherichia coli 10–108 cfu/gStaphylococcus aureus 0.14–0.19 μg toxin/kg

bodyweightPond water, human carrier (cause postharvest contamination)

VirusesHepatitis A Living virus can infect

humansFaecal polluted water,accumulation in shellfishNorwalk virus

AlgaeDinoflagellates E.g. ciguatoxins,

PSP, ASP, DSP, NSP toxinsOpen waters, marine tropicalwaters; accumulation in shellfish (e.g. mussels, oysters)

Parasites Some living parasites can infect humans

Fish and shellfish

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

PATHOGEN BACTERIA ON SEAFOOD

Bacteria

Action model Toxin stability

Minimum dose to infect

infection Toxin forming

Indigenous bacteria

Clostridium botulinum   + Low -Vibrio sp +     HighV. cholerae +     -

V. parahaemoliticus +     (> 106/g)

Aeromonas hydrophila +     NK

Plesiomonas shigelloides +     NKListeria monocytogenes       NK

Non-indigenous

bacteria

Salmonella sp +     < 102

Shigella +     106

E. coli +     101 – 102

Staphylococcus aureus   + High 101 – 103

EKO SUSANTO – DIPONEGORO UNIVERSITY (eko_thp@undip.ac.id)

THANK YOU FOR ATTENTION