MICROBIOLOGICAL QUALITY OF RANDOMLY SELECTED READY-TO-EAT

FOOD ON POULTRY BASED PRODUCT

Prepared by: Muhammad Idris A. Wahid Supervisor: Puan Siti Baizura M. Zain

INTRODUCTION

Objective Significance study Problem statement

INTRODUCTION (CONT.)

Many foods premises such restaurants, cafeterias and even street vendors served ready to eat foods and it becomes a common choice for human in developing countries. People prefer to eat ready to eat foods because it readily cooked, inexpensive, nutritional meals and usually food premises provide a wide varieties of food choices for the customer to eat.

These make ready to eat foods more preferable, but the vendors usually lack of adequate training on basic food hygiene practices and this will give raises concern over the safety of these sold foods.

PROBLEM STATEMENTS

The food price is always be an issues; food sold are quite expensive but the safety level of the food is unknown Food handlers usually lack of adequate training on basic food hygiene practices and this will give raises concern over the safety of these sold foods

Pathogens might be contaminate food during preparation, cooking, reheating and holding period Food borne illness

SIGNIFICANT OF STUDY

Important - Prevent is 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.

SIGNIFICANT OF STUDY (CONT.)

Cared about microbiological hazard in the food we 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.

SIGNIFICANT OF STUDY (CONT.)

Give additional exposure for Food Science and Technology 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.

SIGNIFICANT OF STUDY (CONT.)

When the risk is there, a control measure can be established 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 reduced

or 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.

OBJECTIVE OF STUDY
To isolate microorganism that present in ready to eat cooked dishes

To identify the species of isolated microorganism

Determine the microbiological Q in ready to eat cooked dishes that serves at the cafeteria in order to interpret the risk of these particular food that are present to consumer

READY-TO-EAT FOODS..

Kari

Kicap

Lemak

Rendang

Kurma

Sweet sour

PATHOGENS ASSOCIATED WITH POULTRY

Foodborne human pathogens associate with poultry - Campylobacter spp, Salmonella serotypes, Listeria monocytogenes, Escherichia coli,, Clostridium perfringens, Staphylococcus aureus (Ray and Bhunia, 2007).

SOURCE OF MICROORGANISMS

Air Ingredie nts

Water

Sources of food contamination

Animals Food contact surfaces (McSwane et al.,2004)
Food handlers

FACTORS INFLUENCING MICROBIAL GROWTH

Food

Moisture

Acid

Oxygen

Temperature

Time (Jay, 2000)

Causative Factors of Foodborne Illness 19931997 (by % outbreak)

6% 11% 16%

11% 37%

improper holding temperature poor personel hygiene contaminated equipment inadequate cooking food from unsafe source other

19%

(SOURCE: CENTRES FOR DISEASE CONTROL AND PREVENTION, 2000)

FOODBORNE HUMAN PATHOGENS ASSOCIATE

WITH POULTRY

Campylobacter spp

Salmonella serotypes

Listeria monocytogenes

Escherichia coli

Clostridium perfringens

Staphylococcus aureus

FOODBORNE ILLNESS CAUSE

BY BACTERIA

CAUSATIVE AGENT

TYPE OF ILLNESS

SYMPTOMS ONSET

COMMON FOOD

PREVENTION

Salmonella spp.

Bacterial infection

Nausea, fever, vomiting, abdominal cramps, diarrhea (6 to 48 hours) Nausea, vomiting, abdominal cramps, headaches (2 to 6 hours)

Raw meat: raw poultry, eggs, Properly cook foods, avoid milk, dairy product cross contamination

Staphylococcus aureus

Bacterial intoxication

Food that are prepared with human contact; cooked or processed foods

Wash hands and practice good personal hygiene. Cooking will not inactivate the toxin

(McSwane et al.,2004)

METHODOLOGY
Sample collection Sample preparation Serial dilution

Colonies observation

Incubated

Total Plate Count

Gram staining

Microscopic determination

Biochemical test (confirmation test)

SAMPLE COLLECTION

This study was carried out at Dataran Chendekia UiTM Shah Alam and involved three restaurants and characterised as restaurant A, B and C respectively. All food samples taken for the microbiological test were 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 as lunch hour

SAMPLE COLLECTION (CONT.)

Aseptic technique is use to transfer the sample into a sterile plastic container, using single use sterile utensils The sample collection is based on USDA requirement - Microbiology Laboratory Guidebook 3rd edition, 1998

SAMPLE PREPARATION

The chicken content of each poultry dish are aseptically cut into small pieces to increase the surface area Chicken sample, are weight 25 g and homogenize in stomacher bag with 225 ml of 0.1% peptone water to make tenfold dilution The sample is homogenize for 30 sec in a stomacher Further serial dilution is make as require using 1 ml of the homogenate and 9 ml of 0.1% peptone water
(Meldrum et al., 2006)

SERIAL DILUTION

INCUBATION

Incubation time and temperature are based on aerobic and anaerobic method

Aerobic
The plate will be incubated for 24 48 hours at 37 oC

Anaerobic
The plate will be place inside the anaerobic jar (remove oxygen) and incubated for 24 48 hours at 37 oC

COLONIES OBSERVATION

The total number of bacteria is determined by multiplying the colony count by the dilution factor to yield 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

GRAM STAINING
Inoculate the colony onto the surface of glass specimen

Stain with methyl violet for 20s

Wash off and replace with iodine & leave for 1 min

Wash off with alcohol or acetone & leaving for a few second

Wash with water

Counterstained with safranin

(Collins and Lyne, 1987)

GRAM STAINING

Gram positive staining

Gram negative staining

RESULT OF ENUMERATION FOR AEROBIC AND ANAEROBIC PLATE COUNT FOR RESTAURANT A
Samples CFU/mL Aerobic Lemak Kicap Rendang Kari 2.5 x 106 3.3 x 106 4.0 x 106 Anaerobic Lemak Kicap Rendang Kari 2.1 x 106 2.9 x 106 3.1 x 106 +ve, -ve +ve +ve Round, regular, white opaque, yellow Round, regular, white opaque Round, regular, white opaque +ve, -ve +ve +ve Round, regular, white opaque, yellow Round, regular, white opaque Round, regular, white opaque Gram Morphology

RESULT OF ENUMERATION FOR AEROBIC AND ANAEROBIC PLATE COUNT FOR RESTAURANT B
Samples Kari Sup Lemak CFU/mL Gram Aerobic 3.4 x 106 3.8 x 106 7.5 x 105 +ve +ve, -ve +ve, -ve +ve, -ve Round, regular, white opaque Round, regular, white opaque, yellow Round, regular, white opaque, yellow Round, regular, white opaque, yellow Morphology

Masam manis 2.0 x 104 Anaerobic Kari Sup 2.1 x 106 2.9 x 106

+ve +ve, -ve

Round, regular, white opaque Round, regular, white opaque, yellow

Lemak

4.6 x 105

+ve, -ve
+ve, -ve

Round, regular, white opaque, yellow

Round, regular, white opaque, yellow

Masam manis 4.2 x 103

RESULT OF ENUMERATION FOR AEROBIC AND ANAEROBIC PLATE COUNT FOR RESTAURANT C
Samples CFU/mL Aerobic Kurma Lemak ND ND Gram Morphology

Curry
Rendang

ND
ND Anaerobic

Kurma Lemak Curry Rendang

ND ND ND ND

RESULT OF GRAM STAINING AND MICROSCOPY

EXAMINATION
Restaurant Restaurant A Ayam lemak Ayam kicap Ayam rendang Ayam kari Restaurant B Ayam kari Ayam sup Ayam lemak Ayam masam manis +ve +ve +ve +ve -ve -ve -ve +ve N/A +ve +ve -ve N/A White colony Yellow colony

RESULT OF INOCULATION OF IDENTIFIED COLONIES

ON SPECIFIC AGAR
Agar Mannitol salt agar Colony White opaque +ve Colony found to grow on agar media Observation

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 Simmon citrate Eosine Methylene Blue +ve +ve +ve Slant turn to black in color Orange color appear at the bottom of slant Colony growth appear grey in color Colony found to grow on agar media Colony have a black spot on the centre

Salmonella-Shigella agar +ve

Table : Biochemical test (confirmation test)

RESULT INTERPRETATION

Microbial result will be interpreted in accordance with microbiological criteria based on the guidance documents developed by both the UK Food Protection Agency and FSANZ

These criteria use the present or level of bacterial contamination as an indicator of food safety, and classify prepared foods as being of good, acceptable, unsatisfactory or unacceptable (potentially hazardous) microbiological quality

Test

Microbiological Result

Good

Acceptable
Standard Plate Count

Unsatisfactory

Potentially hazoardous

Category A
Category B Category C

<104
<106 N/A

<105
<107 N/A Indicators

105
107 N/A

N/A
N/A N/A

Enterobacteriaceae E.Coli

<102 <3

102 to <104 3 to <102 Pathogens

104 102

N/A N/A

Coagulase +Ve staphylococci

<102

102 to <103

103 to<104

104 104 104

C.Perfringens

<102

102 to <103

103 to<104

B. Cereus

<102

102 to <103

103 to<104

v. Parahaemolyticus

Not detected in 25 g <3

If detected as per below 3 to <102 102 to<104 104 Detected in 25 g Detected in 25 g L. Monocytogenes

Campylobacter spp Salmonella spp

Not detected in 25 g Not detected in25 g

Food group 1

Not detected in 25 g

Detected in 25 g

Food group 2

Not detected in 25 g

Detected but <102

Detected but <102

102
102

Food group 3

Not detected in 25 g

RESULTS INTERPRETATION (CONT.)

CFU numbers of enterotoxigenic S.aureus (greater than 106 CFU/mL) are needed to produce enough staphylococcal enterotoxin to cause food intoxication (Khalilah, 2007)

RESULTS INTERPRETATION (CONT.)

Based on the result, the white opaque and round shape colonies from aerobic and anaerobic plate are 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 are found gram negative rod shaped bacteria and was suspected to be salmonella spp. confirmed by using TSI, Simmon citrate, Eosine Methylene Blue and SS agar.

CONCLUSION

It can be concluded that the samples taken for the microbiological quality test are contained foodborne pathogens Based on the biochemical test done onto the species of isolated colonies the result showed positive growth of salmonella spp. and staphylococcus aureus bacteria

CONCLUSION (CONT.)

Result was interpreted in accordance with microbiological criteria based on the guidance documents developed by both the UK Food Protection Agency and FSANZ:

The level of staphylococcus aureus and salmonella spp. contamination in samples, can be classified prepared foods as being of unacceptable (potentially hazardous) microbiological quality

RECOMMENDATIONS

Control of cross contamination with proper cleaning and sanitizing Cooked food for sufficient time at the sufficient temperature and keep at proper temperature during hot-holding of foods. Food handlers should always in good health and use good personal hygiene practices.

REFERENCES

Bibek Ray and Arun Bhunia, (2007). Fundamental food microbiology. (4th ed.). C.H. Collins and Patricia M. Lyne, (1987). Microbiological (5th ed.). method.

David McSwane, Nancy R. Rue and Richard Linton, (2004). Essentials of food safety and sanitation. (4th ed.). Pearson Prentice Hall. E. Cardinale, J.D. Perrier gros-Claude, F. Tall, E.F. Gueye and G. Salvat, (2004). Risk factors for contamination of ready-toeat street-vended poultry dishes in Dakar, Senegal. International Journal of Food Microbiology 103 (2005) 157165. G.C. Mead, (2005). Food safety control in the poultry industry. Woodhead Publishing in Food Science and Technology.

REFERENCES

James M. Jay, (2000). Modern Food Microbiology. (6th 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 for ready-to-eat foods: A guide to interpreting microbiological result. R.J. Meldrum, R.M.M. Smith, P. Ellis, J. Garside and on behalf of the Welsh Food microbiological Forum, (2006). Microbiological quality of randomly selected ready-to-eat food sampled between 2003 and 2005 in Wales, UK.

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-away restaurants in United kingdom. Thomas J. Montville and Karl R. Matthew, (2005). Food microbiology: An introduction.