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MICROBIOLOGICAL QUALITY OF RANDOMLY SELECTED READY-TO-EAT FOOD ON POULTRY BASED PRODUCT
Prepared by: Muhammad Idris A. WahidSupervisor: Puan Siti Baizura M. Zain
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INTRODUCTION
Problemstatement
Significancestudy
Objective
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INTRODUCTION (CONT.)
Many foods premises such restaurants, cafeteriasand even street vendors served ready to eat foodsand it becomes a common choice for human indeveloping countries.
People prefer to eat ready to eat foods because itreadily cooked, inexpensive, nutritional meals andusually food premises provide a wide varieties of foodchoices for the customer to eat.
These make ready to eat foods more preferable, butthe vendors usually lack of adequate training on basicfood hygiene practices and this will give raises
concern over the safety of these sold foods.
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P ROBLEM STATEMENTS
The food price is always be an issues; food soldare quite expensive but the safety level of the foodis unknown
Food handlers usually lack of adequate training onbasic food hygiene practices and this will giveraises concern over the safety of these sold foods
Pathogens might be contaminate food duringpreparation, cooking, reheating and holding period Food borne illness
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S IGNIFICANT OF STUDY
Important - Preventis better than cure
A lot of money spend when dealing with food borne illness in order
to run the enforcement, investigating of the outbreaks, sampling of
sample to be analyses at laboratory, cost of the treatment, cost of
the equipment and chemical used etc
The RTE food served at the Cafeteria should be safe for
consumption to avoid any harm to the consumer.
Because of these great consequences it is important to study the
microbiological quality of foods.
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S IGNIFICANT OF STUDY (CONT .)
Cared about microbiological hazard in the foodwe consumed
We should know about the foods we consumed. Its not just only
about their nutritional value, we should also care about the safety
in term of the potential of food borne illness to occur. Hence a
control measure can be taken by the food premises in reducing
the potential hazard.
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S IGNIFICANT OF STUDY (CONT .)
Give additional exposure for Food Science andTechnology student
It will expose student the general knowledge on pathogenic (if
any) microorganisms detection.
As food technology student who dealing most with foods during
food production and processing, it is important for us to have a
basic knowledge about the characteristics of these pathogens (if
any) especially about their optimum growth environment, factors
influences their growth and also how to inhibit the growth of
pathogens in order to reduce their contamination during food
preparation.
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S IGNIFICANT OF STUDY (CONT .)When the risk is there, a control measure can beestablished in order to control the problem
Make it more clear regard the safety of the foods we take, because
some delicious food is not necessarily prepared in the right way.
Once we know the risk is there, a control measure can be establish
in order to overcome the problems and give guideline of good
hygiene practices among the food handlers, that will result in not
only increasing the customer confident, also can raise the
reputation of food premises and the most important is can reducedor eliminated the risk of food borne illness.
The steps taken to keep foods safe from disease-causing bacteria
will also improve the culinary quality and shelf life of the food.
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OBJECTIVE OF STUDY
Determine the microbiological Q
in ready to eat cooked dishesthat serves at the cafeteria inorder to interpret the risk of theseparticular food that are present to
consumer
To isolatemicroorganism thatpresent in ready toeat cooked dishes
To identify thespecies of isolated
microorganism
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READY-TO-EAT FOODS ..
Kari Kicap Lemak
Rendang Kurma Sweet sour
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PATHOGENS ASSOCIATED WITH POULTRY
Foodborne human pathogens associate withpoultry - Campylobacter spp, Salmonella serotypes, Listeria monocytogenes, Escherichia coli,, Clostridium perfringens, Staphylococcus aureus (Ray and Bhunia, 2007).
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S OURCE OF MICROORGANISMS
Sources of foodcontamination
Air
Water
Foodhandlers
Foodcontact
surfaces
Animals
Ingredients
(McSwane et al.,2004)
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FACTORS INFLUENCING MICROBIAL GROWTH
Food
Acid
Temperature
Time
Oxygen
Moisture
(Jay, 2000)
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(SOURCE : C ENTRES FOR DISEASE CONTROL AND PREVENTION , 2000)
37%
19%16%
11%
6% 11%
Causative Factors of Foodborne Illness 1993-1997 (by % outbreak)
improper holdingtemperature
poor personel hygiene
contaminatedequipment
inadequate cooking
food from unsafesource
other
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FOODBORNE HUMAN PATHOGENS ASSOCIATE WITH POULTRY
Campylobacter spp
Salmonella serotypes
Listeria monocytogenes
Escherichia coli Clostridium perfringens
Staphylococcus aureus
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CAUSATIVEAGENT
TYPE OFILLNESS
SYMPTOMS ONSET COMMON FOOD PREVENTION
Salmonella spp. Bacterialinfection Nausea, fever, vomiting,abdominal cramps,diarrhea (6 to 48 hours)
Raw meat: raw poultry, eggs,milk, dairy product Properly cook foods, avoidcross contamination
Staphylococcusaureus
Bacterialintoxication
Nausea, vomiting,abdominal cramps,headaches (2 to 6 hours)
Food that are prepared withhuman contact; cooked orprocessed foods
Wash hands and practice goodpersonal hygiene. Cooking willnot inactivate the toxin
FOODBORNE ILLNESS CAUSE BY BACTERIA
(McSwane et al.,2004)
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METHODOLOGY
Samplecollection
Samplepreparation Serial dilution
Total PlateCountIncubated
Coloniesobservation
Gram staining Microscopicdetermination
Biochemical test(confirmation
test)
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S AMPLE COLLECTION
This study was carried out at Dataran ChendekiaUiTM Shah Alam and involved three restaurants andcharacterised as restaurant A, B and C respectively.
All food samples taken for the microbiological testwere chosen randomly based on poultry origin
The timing of the samples taken was between 11.30
a.m. to 1.30 p.m. at which the time considers aslunch hour
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S AMPLE COLLECTION (CONT .)
Aseptic technique is use to transfer the sample intoa sterile plastic container, using single use sterileutensils
The sample collection is based on USDArequirement - Microbiology Laboratory Guidebook 3 rd edition, 1998
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S AMPLE PREPARATION
The chicken content of each poultry dish areaseptically cut into small pieces to increase thesurface area
Chicken sample, are weight 25 g and homogenizein stomacher bag with 225 ml of 0.1% peptonewater to make tenfold dilution
The sample is homogenize for 30 sec in a
stomacher
Further serial dilution is make as require using 1 mlof the homogenate and 9 ml of 0.1% peptone water
(Meldrum et al., 2006)
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S ERIAL DILUTION
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INCUBATION
Incubation time and temperature are based onaerobic and anaerobic method
Aerobic
The plate will beincubated for 24 48hours at 37 oC
Anaerobic
The plate will be placeinside the anaerobic
jar (remove oxygen)and incubated for 24 48 hours at 37 oC
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COLONIES OBSERVATION
The total number of bacteria is determined bymultiplying the colony count by the dilution factor toyield the number of bacteria per gram of food.
The number of CFU per ml of sample =
The number of colonies The dilution factor of the plate count
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GRAM STAINING
Inoculate the colony onto the surface of
glass specimen
Stain with methyl violet for 20s
Wash off and replace with iodine & leavefor 1 min
Wash off with alcohol or acetone &leaving for a few second
Wash with water
Counterstained with safranin
(Collins and Lyne, 1987)
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GRAM STAINING
Gram negative stainingGram positive staining
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RESULT OF ENUMERATION FOR AEROBIC AND ANAEROBIC P LATE COUNT FOR RESTAURANT A
Samples CFU/mL Gram Morphology Aerobic
Lemak 2.5 x 10 6 +ve, -ve Round, regular, white opaque, yellow
Kicap - - -
Rendang 3.3 x 10 6 +ve Round, regular, white opaque
Kari 4.0 x 10 6 +ve Round, regular, white opaque
Anaerobic
Lemak 2.1 x 10 6 +ve, -ve Round, regular, white opaque, yellow
Kicap - - -RendangKari
2.9 x 10 6
3.1 x 10 6 +ve
+veRound, regular, white opaque
Round, regular, white opaque
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RESULT OF ENUMERATION FOR AEROBIC AND ANAEROBIC P LATE COUNT FOR RESTAURANT B
Samples CFU/mL Gram Morphology
Aerobic
Kari 3.4 x 10 6 +ve Round, regular, white opaque
Sup 3.8 x 10 6 +ve, -ve Round, regular, white opaque, yellow
Lemak 7.5 x 10 5 +ve, -ve Round, regular, white opaque, yellow
Masam manis 2.0 x 10 4 +ve, -ve Round, regular, white opaque, yellow
Anaerobic
Kari 2.1 x 10 6 +ve Round, regular, white opaque
Sup 2.9 x 10 6 +ve, -ve Round, regular, white opaque, yellow
Lemak 4.6 x 10 5 +ve, -ve Round, regular, white opaque, yellow
Masam manis 4.2 x 10 3 +ve, -ve Round, regular, white opaque, yellow
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RESULT OF ENUMERATION FOR AEROBIC AND ANAEROBIC P LATE COUNT FOR RESTAURANT C
Samples CFU/mL Gram MorphologyAerobic
Kurma ND - -Lemak ND - -Curry ND - -Rendang ND - -
Anaerobic
Kurma ND - -Lemak ND - -CurryRendang
NDND
--
--
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RESULT OF GRAM STAINING AND MICROSCOPY EXAMINATION
Restaurant Whitecolony
Yellowcolony
Restaurant A
Ayam lemak +ve -ve
Ayam kicap N/A N/A
Ayam rendang +ve - Ayam kari +ve -
Restaurant B
Ayam kari +ve - Ayam sup +ve -ve
Ayam lemak +ve -ve Ayam masam manis +ve -ve
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Agar Colony ObservationWhite opaque
Mannitol salt agar +ve Colony found to grow on agar media
Color of agar changed from red to yellow
Blood base agar +ve Colony found to grow on agar media Each colony surround with a clear zone Coagulation of blood occurred
Yellow
TSI +ve Slant turn to black in color
Simmon citrate +ve Orange color appear at the bottom of slant
Eosine Methylene Blue +ve Colony growth appear grey in color
Salmonella-Shigella agar +ve Colony found to grow on agar media
Colony have a black spot on the centre
Table : Biochemical test (confirmation test)
RESULT OF INOCULATION OF IDENTIFIED COLONIES ON SPECIFIC AGAR
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RESULT INTERPRETATION
Microbial result will be interpreted in accordancewith microbiological criteria based on the guidancedocuments developed by both the UK FoodProtection Agency and FSANZ
These criteria use the present or level of bacterialcontamination as an indicator of food safety, andclassify prepared foods as being of good,
acceptable, unsatisfactory or unacceptable(potentially hazardous) microbiological quality
Test Microbiological Result
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Good Acceptable Unsatisfactory Potentially hazoardous
Standard Plate Count
Category A
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RESULTS INTERPRETATION (CONT .)
CFU numbers of enterotoxigenic S.aureus (greaterthan 10 6 CFU/mL) are needed to produce enoughstaphylococcal enterotoxin to cause food intoxication(Khalilah, 2007)
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RESULTS INTERPRETATION (CONT .)
Based on the result, the white opaque and roundshape colonies from aerobic and anaerobic plateare characterised as gram positive bacteria,suspected to be a staphylococcus spp. and
confirmed by using two selective different agars;mannitol salt agar and blood base agar.
The yellow, regular, round shape colonies detected
from both aerobic and anaerobic condition arefound gram negative rod shaped bacteria and wassuspected to be salmonella spp. confirmed by usingTSI, Simmon citrate, Eosine Methylene Blue andSS agar.
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CONCLUSION
It can be concluded that the samples taken for themicrobiological quality test are contained foodbornepathogens
Based on the biochemical test done onto the speciesof isolated colonies the result showed positive growthof salmonella spp. and staphylococcus aureus bacteria
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CONCLUSION (CONT .)
Result was interpreted in accordance withmicrobiological criteria based on the guidancedocuments developed by both the UK FoodProtection Agency and FSANZ:
The level of staphylococcus aureus andsalmonella spp. contamination in samples, can beclassified prepared foods as being of unacceptable(potentially hazardous) microbiological quality
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RECOMMENDATIONS
Control of cross contamination with proper cleaningand sanitizing
Cooked food for sufficient time at the sufficienttemperature and keep at proper temperature duringhot-holding of foods.
Food handlers should always in good health anduse good personal hygiene practices.
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REFERENCES Bibek Ray and Arun Bhunia, (2007). Fundamental food
microbiology. (4 th ed.).
C.H. Collins and Patricia M. Lyne, (1987). Microbiological method.(5 th ed.).
David McSwane, Nancy R. Rue and Richard Linton, (2004).
Essentials of food safety and sanitation. (4th
ed.). PearsonPrentice Hall.
E. Cardinale, J.D. Perrier gros-Claude, F. Tall, E.F. Gueye and G.Salvat, (2004). Risk factors for contamination of ready-to-eat street-vended poultry dishes in Dakar, Senegal.
International Journal of Food Microbiology 103 (2005) 157-165.
G.C. Mead, (2005). Food safety control in the poultry industry.Woodhead Publishing in Food Science and Technology.
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REFERENCES James M. Jay, (2000). Modern Food Microbiology. (6 th ed.). Aspen
Publisher.
Khalilah, A.K., (2007). Food Microbiology: a laboratory manual
McLandsborough, L., (2003). Food microbiology laboratory.
NSW Food Authority, (2009). Microbiological quality guide forready-to-eat foods: A guide to interpreting microbiologicalresult.
R.J. Meldrum, R.M.M. Smith, P. Ellis, J. Garside and on behalf ofthe Welsh Food microbiological Forum, (2006).Microbiological quality of randomly selected ready-to-eatfood sampled between 2003 and 2005 in Wales, UK.
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REFERENCES R.J. Meldrum, C.L. little, S. Sagoo, V. Mithani, J. McLauchlin and
E. de pinna, (2009). Assesment of the microbiological safety
of salad vegetables and sauces from kebab take-awayrestaurants in United kingdom.
Thomas J. Montville and Karl R. Matthew, (2005). Foodmicrobiology: An introduction.
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