THE DETECTION OF THE PRESENCE OF MICROORGANISMS IN A BURGER FOOD SAMPLE

D. Antipuesto, C. De la Pea, J. Figueroa, A. Harder,and A. Libo-on

University of Negros Occidental Recoletos
This scientific paper was an attempt to detect the presence of different microorganisms found on a sample of a burger. In conducting the experiment on the presence of different microorganisms found on the food sample, the experiment was done by blending the food sample with peptone water. The food sample was then tested for the presence of coliforms, Escherichia coli, Staphylococcus aureus, and Salmonella. For the detection of the presence of coliforms, the researchers used Lauryl Sulfate Tryptose Broth (LSTB) and Brilliant Green Lactose Bile Broth (BGLBB). For the presence of E. coli, the researchers used Eosin Methylene Blue Agar (EMB). For the detection of Staphylococcus aureus, the researchers used Baird-Parker Agar (BPA). For the detection of Salmonella, the researchers used Rappaport broth first to enrich the sample and the used Brilliant Green Agar (BGA), Salmonella-Shigella Agar (SSA) and Bismuth Sulfite Agar (BSA) to confirm the presence of Salmonella. As the experiment has been conducted, the results showed that the burger sample contained S. aureus due to the positive result exhibited by the sample to the test for S. aureus. Thus, the data obtained in the experiment presents that the burger food sample contained S. aureus.

1. Introduction

Everybody loves burger. Who wouldnt? Because of its juicy patty trapped between two soft buns, sometimes added with cheese, tomatoes, lettuce, and pickles we consider it as one of our favourite fast food of all time. Due to the influence of the Americans we Filipinos learned to love this kind of food. It was first created in America around 1890. Today, many fast food franchises had developed different ways to improve the burgers physical appearance and taste. But, do you know that within this heavenly treat there are microorganisms that can be found in it? These microorganisms can either be pathogenic or non-pathogenic. The

transmitted through air or physical contact. The patty that is made up of grinned beef is prone to microbial inhabitants. Recent studies have shown that burgers can be free from Escherichia coli (E.coli) and other health-risk bacteria. Heat can be one of the factors that can denature proteins that keep these bacteria alive. The objective of this study is to determine the pathogenecity of the burger. Specifically, the study aims to find out (1) the bacteria that are present on the sample, (2) how fast these bacteria can grow and (3) what nutrients can it grow into. The study is relevant to the well-being of humans since that many people are fond of eating this kind of food especially those who are bought on streets. It can also promote

presence of microorganisms is due to unhygienic practices in food processing and preparation. The contaminants can be

awareness about the health-risk factors that can be present on the food that we eat.

salmonellas, always in numbers below 1 per g. The remaining 355 cooked hamburgers were prepared from samples pre-cooked for 10 min

2. Review of Related Literature

at 80 'C. Some were grilled and some fat fried. The total bacterial counts were from 103 to 105 per g, and counts of Enterobacteriaceae below 102 per g. Salmonellae, again in small numbers only, were recovered from 3.5 % of samples. When hamburgers were artificially

S. K. Tamminga, R. R. Beumer AND E. H. Kampelmacher Microbiological studies on Hamburgers November 29, 1980 One hundred and eighty-two raw, 112 precooked and 750 cooked hamburgers

composed mainly of beef or beef and pork were subjected to microbiological

contaminated with Salmonella typhimurium it took 5-5 min on a commercial grill, 2-25 min frying in a frying pan and 1-75 min on a household grill to reliably reduce the

examination. Raw hamburgers gave total bacterial counts from 106 to 108 per
3 3

g,
5

counts

of

salmonella count one hundredfold. This means that at many vending places

Enterobacteriaceae from 10 to 106 per g, ofEscherichia coli from 10 to 10 , of group D streptococci from 10
2 4

hamburgers are often cooked for too short a time. D-values were determined for S.

to

10 ,

of

Staphylococcus aureus from 3 to 102 and of Clostridium perfringens less than 10 bacteria per g. Of the samples, 32 % contained salmonellas; the highest most probable number was 102 per g but most estimates were below 1 per g. Corresponding figures for the pre-cooked samples were 2-3 log cycles lower, and only one sample contained salmonella. Yersinia enterocolitica was not isolated from any raw or pre-cooked sample. Three hundred and ninety-five of the cooked hamburgers were prepared by grilling raw hamburgers for between 2 and 5-5 min. These gave total bacterial counts from 105 to 107 per g, and counts of Enterobacteriaceae from 102 to 105 per g. Of the samples, 9-4 % contained

typhimurium in hamburger meat at 50, 55, 60, 65 and 70 'C, these values were 7-1, 5-1, 1-2, 0 9 and 0-6 min respectively. It can be concluded that the heating action in the centre of the hamburgers will take place more slowly than in the hamburger as a whole, and that the time between cooking and

consumption is very important in reducing the microbial load to acceptable levels. Pre-cooking (10 min at 80 C in a water bath) gives a reduction in the numbers of salmonella of about 4 x 103, after which cooking gives a further reduction as

mentioned above.

These experiments show, in agreement with other authors, that raw minced meat is contaminated by a wide range of microorganisms, often present in large numbers. The level of micro-organisms, especially that of hygienic indicator organisms, draws

refrigeration. Few (30%) vendors displayed their food handler's badge. Most wore hair restraints. Doneness of beef patties was

judged solely by appearance. In all cases, the assemblage and serving of hamburgers was facilitated by the use of tongs. No to low microbial risk was noted in samples for 81.5% Salmonella, 71.4% Staphylococcus aureus

attention, yet again, to measures which might be taken to counteract the consequences of the unavoidable original contamination of meat during slaughtering, such as cooling, limitation of storage period and regular cleaning and disinfection of apparatus and utensils. The value of the indicator organisms in judging the possibility of the presence of pathogenic organisms is very limited, for example when Enterobacteriaceae counts are relatively low, a considerable percentage of samples may still contain salmonella.

and 96.3% Escherichia coli of samples. The presence of foodborne pathogens posed some potential risks to consumers. The presence of microbial pathogens in some beef patties could pose a potential health concern to the consuming public. Overall the study highlighted the need for effective communication on microbiological food risks, proper instruction and supervision in food handling procedures, greater

consumer/vendor education on transmission N. Badrie, A. Joseph and A. Chen An Observational Study of Food Safety Practices by Street Vendors and of enteric food - borne diseases and food safety risks, how safely to cook and serve foods and more vigilant monitoring by

Microbiological Quality of Street-Purchased Hamburger Beef Patties in Trinidad, West Indies There has been minimal study on the food safety practices by street-food vendors in Trinidad, West Indies and little is known on the microbiological quality of street-

food inspectors and control staff.

C.L. Little, I. A. Gillespie and R. T. Mitchell Microbiological examination of ready-to-eat burgers sampled anonymously at the point of sale in the United Kingdom.December 4, 2001 During May and June 1999 a microbiological study of ready-to-eat burgers purchased anonymously from burger outlets (combined take-away and burger restaurants, take-awayonly fixed premises, mobile vendors,

purchased

hamburgers.

Microbiological

analysis was conducted on 27 beef patties. Most (80.1%) vendors sold hamburgers from readily removable units. Only 10% of vending units had accessed to running water and

temporary stalls and other burger outlets)

was undertaken. The intention was to determine the microbiological quality of ready-to-eat burgers as purchased by

materials: 25 grams of the burger sample, digital balance, 225ml of peptone water, a blender, Erlenmeyer flasks, cotton plugs, aluminium foils, petri dishes, micropipettes, serological pipettes, bulbs, liters of distilled water, dozens of test tubes, test tube racks, and screw cap tubes. The mediums used to determine the bacteria present in the food sample were Rappaport, LSTB, BGA, BGLBB, BPA, EMB, TSA, SSA, TSIA, and LIA.

customers of take-away premises and to ascertain, where information was available, whether the Chief Medical Officer's advice on cooking burgers was being followed.

Examination of 3,128 ready-to-eat burgers found that 2,868 (92%) were of acceptable quality and 260 (8%) were of unsatisfactory quality. Unsatisfactory results were mostly due to high aerobic colony counts (ACCs). Salmonella spp., Campylobacter spp. and Escherichia coli O157 were not detected in any of the samples examined. Acceptable microbiological quality of ready-to-eat

3.2 Preparation The test required only a small amount from the specimen. Specifically, 25 grams of the burger was needed. We carefully weighed 25 grams of the burger on a digital balance. We needed it to be shredded in very fine pieces so we placed it in a blender with 225 ml of peptone water to make 250 ml of the mixture. The mixture was then placed in an Erlenmeyer flask, covered with a cotton plug and aluminium foil. In order for microorganisms to grow, the mixture was placed in an incubator, where the temperature and environment was fit for their growth and multiplication. A serial dilution up to 10-2 was prepared for the

burgers was associated with outlets, such as combined take-away and burger restaurants and in particular national franchise outlets, which had management food hygiene training and hazard analysis in place. Poor

microbiological quality was associated with undercooking and local outlets as indicated by Local Authority Inspectors' Consumers at Risk scores.

3. Methodology

different

test

for

the

detection

of

microorganisms present in the food sample. 3.1 Materials We were assigned to conduct an experiment on the presence of 3.3Detection of Coliforms For the detection of coliforms present in the burger sample, we used three Durham tubes with 10 mL Lauryl Sulfate Tryptose Broth (LSTB) containing 1mL of three successive

microorganisms in a burger of a popular local fast food chain in Negros Occidental. To accomplish this test, we used these following

dilutions. The tubes were incubated at 35C for 24 hours. Confirmatory test for the presence of Coliforms was done by

BGA plate will produce a colorless, pink to fuschia, tanslucent to opaque, with

surrounding medium pink to red. The positive result for SSA is colorless to pale pink, opaque, transparent or translucent colonies, with or without black center. The positive result for BSA is brown, black, other times with metallic sheen colonies.

transferring a loopful of each gas positive tubes of LSTB to a tube of Brilliant Green Lactose Bile Broth (BGLBB) and streak on Eosin Methylene Blue Agar (EMB). The tubes and the plates were incubated at 35C for 24 hours. For the test for Escherichia coli, the EMB plates with a positive result for E. coli will produce a green sheen on the surface of the plate.

4. Results and Discussions

This

chapter

presents,

analyzes

and

interprets the data gathered from the experimentation of a burger food sample. The

3.4 Detection of Staphylococcus aureus For the detection of Staphylococcus aureus, transfer 0.1mL of two serial dilutions to a plate of Baird-Parker Agar (BPA). A positive result of the presence of S. aureus will show a black and convex colonies surrounded by a clear zone.

purpose of this study is to detect different microorganisms present in burger food sample. The following statements will provide the data of the experiment conducted to test the hypothesis formulated in this study.

Table 1. Detection of Coliforms using Lauryl Sulfate Tryptose Broth (LSTB)

3.5 Detection of Salmonella For the detection of Salmonella, first the food sample should be enriched by

Tube # T1 (10 ) T2 (10 ) T3 (10 )

-2 -1 0

Result Gas formation Gas formation Gas formation

Interpretation Positive Positive Positive

transferring 1mL of three serial dilutions of the food sample to Rappaport broth. Incubate at 35C for 24 hours. After the incubation, the researchers streaked a loopful from the enrichment medium to the following

differential media: Brilliant Green Agar (BGA), Salmonella-Shigella Agar (SSA) and Bismuth Sulfite Agar (BSA). Incubate at 35C for 24 hours. The positive result of Salmonella in

Table 1.2. Detection of Coliforms using Brilliant Green Lactose Bile Broth (BGLBB)
Tube # T1 (10 )
-1 0

Table 3. Detection of Salmonella enrichment using Rappaport broth

Result Blurry with gas formation

Interpretation Positive

Tube # T1 (100) T2 (10-1) T3 (10-2)

Result Blurry medium Blurry medium Blurry medium

Interpretation Growth

T2 (10 )
-2

Blurry with gas formation

Positive

Growth

T3 (10 )

Blurry with gas formation

Positive

Growth

Table 1.3 Presence of Escherichia coli using Eosin Methylene Blue Agar (EMB) Plate # P1 (10 ) P2 (10 ) P3 (10-2)
-1 0

Table 3.1.. Detection of Salmonella using Brilliant Green Agar (BGA) Tube # P1 (100) P2 (10-1) P3 (10-2) Result Brownish colony Brownish colony Brownish colony Interpretation Negative Negative Negative

Result Brown colonies Brown colonies Brown colonies

Interpretation Negative Negative Negative

Table 2. Detection of Staphylococcus aureus using Baird-Parker Agar (BPA) Tube # P1 (10 )
0

Result Brown with black center, surrounding medium is pink

Interpretation

Table 3.2 Detection of Salmonella using Salmonella-Shigella Agar (SSA)

Positive

Tube # P1 (100)

Result Dark brown surface

Interpretation Negative

P2 (10-1)

Brown with black center, surrounding medium is pink Positive

P2 (10 )

-1

Dark brown surface

Negative

P3 (10-2)

Light brown surface

Negative

P3 (10-2)

Brown with black center, surrounding medium is pink Positive

Table 3.3 Detection of Salmonella using Bismuth Sulfite Agar (BSA) Tube # P1 (100) Result Brown surface P2 (10-1) Brown surface P3 (10-2) Brown surface Positive Positive Interpretation Positive

6. References 1. www.mendeley.com 2. www.internetjfs.org 3. www.ncbi.nlm.nih.gov

A physiological test was supposed to be done with the positive result for BSA using Triple Sugar Iron Agar (TSAI) and Lysine Iron Agar (LAI). These tests were not done due to the negative result of the detection of Salmonella using BSA. 5. Conclusion In conclusion to this paper based on the results of the experiment on the detection of the presence of microorganisms in a burger food sample, the researchers have concluded that Staphylococcus aureus is present in the food sample due to the positive result it showed in the test for the detection of S. aureus in the burger food sample. The researchers also concluded that S. aureus has been found on the food sample due to reasons such as, unhygienic practices in food processing and preparation, the transmission of contaminants through air or physical contact and the main ingredient, ground beef, which is prone to microbial inhabitants