LABORATORY REPORT

FOOD SCIENCE AND NUTRITION (SBK3033)

TITLE FOOD MICROBIOLOGY

DATE OF EXPERIMENT 25/5/2023

GROUP A

LECTURER NAME DR. SUZITA RAMLI

NAME MATRIC NO.

NURATIRAH BINTI MOKHTAR D20221101707

NORAIN FARZANA BINTI SADZALI D20221101730

NURUL FATIHAH BINTI NURHADI D20221101733

NINA SHARLINA BINTI SELAMAT D20221101736

NUR ALIA SYAFIQHA BINTI MOHAMAD D20221101737

NASIR

NUR JUMAATUL NAZAWIYAH BINTI D20221101746

SAZLIN HELMI

NIK NUR SYAHIRAH BINTI NIK DANYA D20221101748

EXPERIMENT 1 :MICROBIAL CONTAMINATION
OBJECTIVES
1.To learn the spread-plate count technique.
2.To estimate the number of bacterial in food samples at different storage time..
3.To understand the preservative nature of high osmotic pressure.

INTRODUCTION
Humans have struggled with microbial food contamination throughout history. Hominids
consumed incredibly polluted food during the majority of the five million years of our
development, including food that was coated in dirt, nibbled on by rats and mice, infected with
maggots and different microorganisms, rotting to varying degrees, etc. It is obvious that we are
not as sensitive as we may think. On the other hand, rotting and rotten food definitely caused
discomfort in many of our predecessors and undoubtedly caused discomfort in many more. Only
the third generation has benefited from secure and safe food preservation techniques. The
Northwest area has had more than its fair share of catastrophic food-borne bacterial illness in
recent years, which is brought on by a new.

EXPERIMENT 1(A). BACTERIAL COUNTS IN VARIOUSLY STORED MEAT

MATERIALS AND EQUIPMENTS

● Bread
● Nutrient agar
● Sterile water
● Media containing a high concentration of salt or sugar
● Hockey stick
● Plate
● Pipette
METHODS
1. The food sample, which is bread, was prepared by adding (20 g + 180 ml of sterile water)
into stomacher bag. The sample was stomached by using stomacher machine for 3 min at
250 rpm or until the sample was blended properly.
2. The dilution solution (9 ml in each test tube) was prepared.
3. 1 ml of blend sample was took out and was added to the first test tube (9 ml sterile water/
nutrient broth)
4. Dilution was made based on the following: (10-2, 10-3, 10-4, 10-5, 10 -6).
5. 0.1 ml of the sample from each test tube was added to the centre of the plate.
6. The glass spreading rod was removed from the ethanol breakers and passed through the
flame. Do not hold it in the flame. The ethanol will catch fire. The beaker was allowed to
burn off away from the beaker. The beaker was holded for about 30 seconds until it cools.
7. The plates were successively spread. Each dilution was duplicated. Do not flame the
spreading rod between plates. The plates were allowed to sit at room temperature until the
liquid soaks into the agar, then placed in the incubator (refer Figure 2).
8. The culture was incubated at 37oC until the next lab and then the number of colonies per
plate was counted. Plates with greater than 300 isolated colonies were labelled TNTC (Too
Numerous To Count) and below 30 are labelled as TFTC (Too Few Too Count). The
number of bacteria in the original sample was calculated using this formula.

Count (CFU/ml or CFU/G)= (average number of colony)x[(Dilution factor ×volume plated)]

RESULT

SAMPLE TOTAL PLATE COUNT

10-2 too numerous to count

10-3 to numerous to count 434

10-4 180

10-5 too few to count 20

10-6 too few to count

 Result of experiment 1(a)

DISCUSSION
Based on the theory, there is a relation between dilution and count. The dilution that has the size
of the test tube was the original size, 10-1 , 10-2 ,10-3 and so on while count is the average of
colonies on the plates of a specific dilution factor. The agar provides a solid growth surface for
the bacteria, upon which bacteria reproduce until the distinctive lumps of cells that we call
colonies form. Then, based on the result obtained also the same. There is bacteria found for
calculation of colony-forming units. For the 10-2 and 10-3 agar plate, the bacteria is too numerous
to count while for 10-4 , there is 180 bacteria growth in the agar plate.Meanwhile, 10-5 and 10-6
bacteria were too few to count on the agar plate. The number of counts similarly reduced as the
number of dilutions decreased.
EXPERIMENT 1(B). EFFECT OF HIGH OSMOTIC CONDITIONS ON THE GROWTH
OF BACTERIA

MATERIALS AND EQUIPMENTS

▪ Broth
▪ Food samples
▪ Microscope

METHOD
1. 4 broth tubes per pair with the following concentrations of salt or sugar were obtained: Salt- 0,
1%, 5%, 15% or sugar- 0, 5%, 25%, 50% (percentage by weight; i.e. for a 50% sugar solution
add the dried medium (for 100 ml of medium) to 50 g of sucrose and sufficient water added to
bring the solution to 100 ml final volume).
2. Each tube was labelled.
3. Each tube inoculated with 10 drops of the original meat suspension (allow the large chunks of
meat to settle out first).
4. Each tube were well mixed.
5. All tubes were Incubated as described by the instructor.
6. The amount of growth in each tube following incubation were estimated, assuming that the
growth in the control tube was + and no growth is (-).
7. These results were recorded in the table below.

RESULTS
Table 1
Osmotic Agent Percent Growth
0 0 +
Sugar 5 +
Sugar 25 +
Sugar 50 +
Salt 1 +
Salt 5 +
Salt 15 +
 Result of experiment 1 (b)

DISCUSSION
Based on this experiment, the foreign matter in the solution represents the growth of the bacteria.
All of the tubes are suitable for bacterial growth. The most foreign matter was shown in 0%
solution that bacteria can grow faster. The least foreign matter is found in salt 15% that the
bacteria cannot grow any further. There are many factors might affect the growth of bacteria such
as temperature, pH, osmotic pressure.

QUESTIONS

1. Explain why cuts of meat like steak are not considered as dangerous for bacterial
infection as hamburger.
Steak are considered less dangerous than hamburger because when grilling a steak, all
exposed parts are heated until the bacteria is killed. Meanwhile the procedure of mincing
meat ensure that the outside end up on the inside, and the bacteria spreads throughout the
entire patty.
 2. What are the symptoms of E. coli, O157:H7 infection?
Symptoms of E. coli, O157:H7 infection are nausea, diarrhea and fatigue.

3. What is the treatment for E. coli, O157:H7 infection?

E. coli O157 infection has no specific treatment.Most sick individuals can be looked after
at home and will recover without the need for medical attention.Even antibiotics are not
helpful for treating E. coli O157:H7 infection. Antidiarrheal agents should not be used
either but drinking plenty of liquids is crucial since diarrhoea can cause dehydration.

4. How can you tell if your hamburger is cooked properly?

It is essential to use a food thermometer to know whether the hamburger is cooked well
or not.This can prevent undercooking,verify that food has reached a safe minimum
internal temperature and consequently prevent foodborne illness.For hamburger at 160
°F, a safely cooked patty may look brown, pink, or some variation of brown or pink.You
can place the thermometer probe sideways or at an angle into thin hamburgers when
grilling them.

CONCLUSION

In this experiment , we determine that the number of counts similarly reduced as the number of
dilution decreased using the provided formula as the theory . The fungi were growing
successfully due to the factors of presence of water , suitable pH and temperature of the bread .

In the experiment of 1(B) , The foreign matter in the solution represents the growth of the
bacteria . The most foreign matter was shown in 0% solution that bacteria can grow faster . This
experiment shows that bacteria grow in the presence of water and oxygen . osmosis is powered
by the potential energy of a concentration gradient and does not require the expenditure of
metabolic energy . While water molecules are small enough to pass through the cytoplasmic
membrane bacteria . Most of the bacteria require an isotonic solution environment for optimum
growth .

In addition , the least foreign matter found in salt 15% that the bacteria cannot grow any further .
When there are high concentrations outside of a bacterial cell , water from inside the bacteria
diffuses out of the cell in order to reach equilibrium and equalize the salt concentration . When
the bacteria cells lose all of their water it causes the loss of the cell’s structure and the cells
dehydrates which leads to the cell death .
REFERENCES

1. Ahern, H. (n.d.-b). Bacteriological Culture Methods. Pressbooks.

https://milnepublishing.geneseo.edu/suny-microbiology-lab/chapter/bacteriological-cultur

e-methods/#:~:text=Agar%2C%20which%20is%20a%20polysaccharide,that%20we%20

call%20colonies%20form.

2. Joan Petersen & Susan McLaughlin Introduction to enumeration of bacteria. (2021, June 2).

Biology LibreTexts.

https://bio.libretexts.org/Courses/North_Carolina_State_University/MB352_General_Mic

robiology_Labo

3. Melissa Mayer The Effects of Salt Concentrations on Bacterial Growth. (2018, May 7).

https://sciencing.com/effects-salt-concentration-bacterial-growth-5924409.html