UNIVERSITI TEKNOLOGI MARA

FAKULTI KEJURUTERAAN KIMIA

FOOD PRESERVATION TECHNOLOGY
(CBE648)
NAME : NUR ILHAM BINTI ZAINUDDIN
STUDENT ID : 2016250248
GROUP : EH2207C
: DETERMINATION OF TOTAL AEROBIC
EXPERIMENT TITLE PLATE COUNT ON FRESH AND SPOILED
FOOD
DATE PERFORMED : 11 OCTOBER 2018

SEMESTER :7

No. Title Allocated Marks (%) Marks

1 Abstract 10
2 Introduction 15
3 Literature Review 15
4 Methodology 10
5 Results & Discussion 30
6 Conclusion & Recommendations 10
7 References 5
8 Appendices 5
TOTAL MARKS 100

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Date :
1. Abstract

The microorganism is present in our environment that cause an effect to foods by their
biochemical activities carried under the ideal growth conditions and cannot be seen by eyes.
It is becoming challenged and only with the assistance of the microscope, we can observe the
presence of microorganism. In this experiment, aerobic plate counter (APC) is used to
observe and calculate the number of colony count forms on the spoiled and fresh sample of
bread and milk. The spoiled bread has the highest number of colony count, which are 999
because it has been exposed to air for a long period of time. However, because of some
limitations of APC, no accumulation of the colony does not indicate that there are no
pathogen has affected the sample.
2. Introduction

2.1 Overview

Food spoilage is a process that causing foods not suitable for human consumption due
to changes in its characteristics such as texture, smell, taste or appearance and maybe
certain of foods have formed of microorganism on it. Shelf life of food is the duration
of food during its stable and retains quantities {Rawat, 2015}. The spoilage
microorganisms rate can be influenced by factors like moisture content, appropriate
temperature, pH and nutrients to grow in the food.

2.2 Problem Statement

Microorganism present in our environment which will affect the foods by biochemical
activities carried under the ideal growth conditions. Human eyes can not resolve
microbes that are too small to be seen which is less than 0.1 mm in diameter without
any help of a microscope {Fung, 2009}. It becomes challenging to find a way to
prevent the activities and growth of microorganism in food for microbiologists,
engineer and technologist {Rawat, 2015}. Thus, this experiment is conducted to
study the total aerobic count (TAC) to determine the number of in the fresh and spoil
milk and bread using streaking process.

2.3 Objective

To determine total aerobic plate count on fresh and spoiled milk and bread by using
streaking process.
3. Literature Review

The result in chemical, physical, biochemical, mechanical, organoleptical, microbiological

and others are contributing to the food spoiled. However, it is very hard to classify food
spoilage especially when the culture itself has various ideas on constitutes edible or nonedible
food such as cheese. With some alteration and adoption process, the taste of the foods can be
changed and people tend to love food from another culture. Food can be considered as fresh
or not fresh is by depending on their conditions. For instance, for fresh fruits they are usually
juicy and crispy compared to the rotten fruit; fresh vegetable is usually newly harvested and
firm while vegetable not fresh is stored and wilted; and fresh bread is moist and fresh
compare to spoil bread {Fung, 2009}. As known, milk contains lactobacillus bacteria will use
lactose to gain its energy and produce lactic acid, which causing the spoilage of milk and
makes the milk become sour {Y*, 2017}. Microbes can grow slowly or inactive at low
temperature, which with the refrigeration method, it can prolong the lag phase and the
microbe growth rate will decrease and it also needs high water activity thus keeping products
in dry condition will preserve them.

For the analysis of food spoilage, it is crucial to have a proper sampling procedure which the
microbiological need to be grouped as solid, liquid, surface or air samples in order to have
more useful data analysis. The conventional ‘standard plate count method’ has been used for
100 years ago where it includes media preparation, sample dilution, sample plating with
general selective agar, plate incubation at specific temperature and colony counters after 48
hours in incubators. There are various manipulative parameters used such as volume of
sample to be plated, the selection of agar, incubation time, temperature and gaseous
environments and others. {Fung, 2009}

The aerobic plate counter (APC) also known as aerobic colony count is a test apply to
determine the level of microorganism in food when it being mixed with agar containing
nutrients. APC is used in evaluating the food quality which depends on the number of
bacteria as an indicator of poor sanitation of equipment and utensils or problems with control
of the process or ingredients. For example, APC used to assess the adequacy of moisture
control during the drying process in dried food where for meats, to check the condition of
incoming carcasses to potentially identify suppliers who prove those with excessively high
counts. APC can be performed in several ways, such as using pour plate, spread plate or
spiral plating techniques. In findings the dilution of countable colony, volumes of test
solution dilutions is fixed when using spread plates and pour plate while for spiral plate,
liquid extract of foods is applied in a spiral track to the surface of rotating agar plate. Colony
counter usually used with a spiral plate to determine CFU/g {Lihua, 2008}.
 4. Methodology

For liquid sample;

I. Fresh and spoil liquid sample was prepared.

II. Gloves were worn and sanitized with hand sanitizer to avoid contamination.
III. Petri dish was labelled on the bottom plate to preserve area to observe the growth.
IV. Fresh liquid samples were poured into petri dishes.
V. The sterile L-shape was dipped into the sample and streak in 360
VI. Then, the sterile L-shape was dipped in distilled water to remove a liquid sample
trace.
VII. Step 3-6 was repeated for spoiled liquid sample.
VIII. The petri dish was sealed with parafilm tape after streaking.
IX. The petri dishes were kept inverted and incubated in an incubator for 20 hours at
37 C.
X. The next 20 hours, the petri dishes were taken out from the incubator and the number
of colonies was counted by using a colony counter.
XI. The amount of colony form was observed and recorded.

For solid sample;

I. Fresh and spoil bread sample was prepared.

I. A sterile cotton swab used to take a bit sample of fresh bread.
II. The sample of bread was streaked on the surface of the agar.
III. Step 2-3 was repeated for spoiled bread sample.
IV. The petri dish was sealed with parafilm tape after streaking.
V. The petri dishes were kept inverted and incubated in an incubator for 20 hours at
37 C.
VI. The next 20 hours, the petri dishes were taken out from the incubator and the number
of colonies was counted by using a colony counter.
VII. The amount of colony form was observed and recorded.
5. Result and Discussion

Aerobic Plate Counter (APC)

Food Sample
Fresh Spoil

Milk 0 208

Bread 598 999

Table 5.1: Result of Aerobic Plate Counter for fresh and spoil samples.

Figure 5.1: Milk sample (fresh and spoil)

Figure 5.2: Bread sample (fresh and spoil)

 In this experiment, aerobic plate count method (APC) is used to calculate the number
of microorganisms. Fresh bread shows slightly higher of colony count comparison to spoiled
milk. From here, we can say that fresh bread has higher colony count due to poor in proper
sanitation and has been infected before by microorganism. Even though the lower value of
APC recorded, it does not count that the food as microbes free because APC is a poor
indicator for microbes present. This is because this method is only taken into account of the
visible colony, need a longer period of time for colony to grow and can cause some error in
the process of counting colony if the colony form is a very large number.

Four plates of samples containing both fresh and spoil food is prepared. The samples
used are bread and milk where the colony count is observed and counted for all of the agar
plates. The spoiled bread shows highest colony count, which is 999 number of colony
compare to the other samples; followed by fresh bread and spoiled milk which are 598 and
208 respectively. Fresh milk contains no accumulation of bacteria in 20 hours of incubation
process.

Moreover, each of the plates has different conditions after the incubation process
where it is depending on the period of the sample exposed to air. This streaking process is
suitable for for heat sensitive bacteria, but also can cause bacteria to be killed when exposed
to oxygen from the surrounding. Spoiled bread has a highest colony count because it exposed
to air in a long time and more colony that’s been killed embedded in the sample. Fresh milk
has a lowest colony count because it exposed to air in shortest time.

As known, some of spoilage organisms need oxygen, some of them are killed by
oxygen and still others are facultative. To solve this problem, the change of plate to modifies
atmosphere packaging become an option which can retard and prevent growth of microbe.
These procedures must be assessed for compatibility with different foods so that there are no
significant organoleptic changes in the foods caused by the treatment or preservative.
6. Conclusion and Recommendation

 Conclusion

All in all, it is concluded that APC is a poor indicator of microbial present because of
some limitations in that process. Lower value of APC does not indicate that the food is
microbial free. However, it can be said that the value of of colony count depends on the
condition of the plates. The spoiled food sample shows a higher number of colony count
rather than fresh food sample and fresh food sample that has high value of the colony because
of the poor sanitation while conducting the experiment.

 Recommendations

1. Make sure to wear gloves and sanitize with hand sanitizer to avoid any contamination of
microbial.
2. Use the suitable method to calculate the colony count to avoid error in counting the
colony.
3. For fresh milk that has no accumulation within 20 hours of incubation process, it is
advisable to incubate the sample for longer time.
4. Streak the agar one time only to avoid a lot of layers of food sample and make it difficult
for colony counter later.
5. Open the packaging of fresh food right before starting the experiments to avoid
contamination of food samples by bacteria.
7. References

 Aerobic Plate Count – Murray Brown Labs. (n.d.). Retrieved from http://mb-
labs.com/resources/aerobic-plate-count/
 Fung, D. Y. C. (2009). Food Spoilage, Preservation and Quality Control. Applied
Microbiology: Agro/Food.
 Lihua, F., & Jun, S. (2008). Microbial quality assessment methods for fresh-cut
fruits and vegetables. Stewart Postharvest Review, 4(3), 1-9.
doi:10.2212/spr.2008.3.10
 Rawat, S. (2015). Food Spoilage: Microorganisms and their prevention. Asian
Journal of Plant Science and Research, 47-56.
 Y, S. (2017). A Short Review on Milk Spoilage. Research and Reviews: Journal
of Food and Dairy Technology.
8. Appendices

Figure 8.1: Colony counter

Figure 8.2: Seal the plate with parafilm

Figure 8.3: Pour the fresh milk sample on agar

Figure 8.4: Streak the agar with sterile L-shape

Figure 8.5: Streak the agar with sterile cotton swab

Figure 8.6: Swab a bit of spoiled bread sample