Experiment 6: Isolation of Microorganisms from Environmental

Samples

Group 9:
So, Ignacius De Charles T.
Tria, Allysa Hannah D.J.
Villafuerte, Arman Joseph F.
Villasenor, Jose Manuel J.
Wahab, Dana Trisha N.

Department of Biology, College of Science, University of Santo Tomas

Abstract
The aim of the experiment is to isolate microorganisms from different environmental sources, specifically
airborne microorganisms and microorganisms from human skin. Exposing agar plates in an enclosed
area in UST carpark was done in order to obtain airborne microorganisms. On the other hand, microbes
from human skin were obtained through swabbing. The study also aims to affirm the efficiency of alcogel
brands in killing microbes (99.9%). This was done by applying Green Cross alcogel on one thumb and
doing a thumbprint on an agar plate. Another agar plate contained a thumbprint without application of any
alcogel. A plate count is done 24 hours after the experiment. Plate counts from agar plates exposed in the
UST carpark yielded 3 and 4 colonies while plate counts for the nose swab were both too many to count
(TNTC). Results from the thumbprint with alcogel plate counts yielded 40 and 1 while the thumbprint
without alcogel yielded 10 and 2. Results show that one alcogel plate was able to reduce microbial count
to 50%. However, the other alcogel plate yielded quadruple the microbial colonies of its counterpart.

Introduction
Microorganisms are living organisms that are immensely small to be seen by the naked
eye because they are microscopic. They are 1 millimeter or less in diameter and can be found
distributed everywhere on our environment. Also, their existence is ubiquitous because they can
thrive as long as there are organic and inorganic compounds that are present in the
environment.

Microbial growth on the environment depends on the tolerance of microbes to the

conditions of the habitat and the availability of nutrients. Most of the microbes can thrive in
oligotrophic environments in which the availability of nutrients is scarce. The evolution of
microbes to grow on famine is due to their adapting and survival response to the constantly
changing harsh environments over time.
 Culture media is a solid (Agar) or a liquid (Broth) artificial preparation used in the
laboratory to grow and culture microorganisms. There are different kinds of medium used per
microbe. Every species has nutritional requirements of the medium for it to effectively grow. In
the laboratory, microbes are isolated and identified through culturing in the medium. Also, they
are tested for antibiotic sensitivities, water and food analysis and for other industrial
microbiology.

This experiment aims to isolate the microorganisms gathered from environmental

sources and to enumerate the different microbial species present in the given environment.

Methodology
In the experiment, half strength of Nutrient Agar (NA) and half strength of Potato
Dextrose Agar (PDA) were measured and prepared for the culture medium of the microbes.
Nutrient Agar and Potato Dextrose Agar were poured onto three plates each and were allowed
to cool and solidify. Then, the plates were subjected to autoclave sterilization to prevent
contamination in the medium for the preparation of the gathering of pure microbes. Airborne
microorganisms were obtained through exposing one Nutrient Agar and one Potato Dextrose
Agar inside the Multiventure room in carpark for two minutes. Microorganisms on human skin
were obtained through the use of swabbing the autoclaved cotton swab on top of a human body
part. Microorganisms existence tested on hands with and without the application of Green
Cross alcogel. One thumb was applied with alcogel while the other thumb was not. After drying,
both thumbs were imprinted into strength Nutrient Agar and strength Potato Dextrose Agar.
After all the microorganisms acquired, all the plates were incubated at room temperature for one
day. Thereafter, microbial growths on the culture media are observed and counted.

Results
*Pictures were taken a few days after plate count. As such, colonies might have already proliferated.

A. Air Exposure (Multi-venture, Ground Floor, Carpark, UST)

 Nutrient Agar: 4 colonies Potato Dextrose Agar: 3 colonies

B. Nose Swab

Nutrient Agar: TNTC Potato Dextrose Agar: TNTC

C. Alcogel (Green Cross)

*Photos taken a week after experiment. Colonies have already proliferated.

 Nutrient Agar (Without alcogel on left half of plate, with alcogel on right half)
Without alcogel: 10 With alcogel: 40

Potato Dextrose Agar (Without alcogel on left half of plate, with alcogel on right
half)
Without alcogel: 2 With alcogel: 1
Tabulated Results (All groups)
 A. Air Exposure

Group number Area NA plate count PDA plate count

1 Female Restroom
2 Cafeteria
3 Gen. Ed. Faculty
4 ATM machine
5 Stairs
6 Mens Restroom
7 Autoclave
8 Locker
9 Multi-venture Carpark
4 0
2 2
4 1
21 1
11 12
0 0
2 1
2 7
3 4

B. Human skin
Group number Area NA plate count PDA plate count
1 Pimple 0 0
2 Ears 21 9
3 Between fingers 61 24
4 Sole of feet TNTC 10
5 Behind the knee 22 0
6 Under tongue 0 0
7 Bikini area 66 134
8 Belly button TNTC TNTC
9 On nose TNTC TNTC

C. Alcogel

Group Number Without Alcogel With Alcogel

1 10 41
2 10, 0 22, 25
3 TNTC, 42 0, 0
4 0, 1 2, 9
5 8 1
6 0 0
7 16, 2 81, 0
8 6, 8 0, 0
9 10, 2 40, 1

Discussion

According to Thiel (1999), factors that contribute to the growth of microbes include
temperature, pH, oxygen, salt concentration, and available nutrients. While most bacteria thrive
best at temperatures of 25-40 degrees Celsius, some bacteria have proven its adaptability to
more extreme cases such has high temperatures and acidities. As such, the growth factor for
each microorganism is unique.
 In order to maintain growth and metabolic functions, microorganisms need to attain
certain basic nutrients from its environment. These include water, nitrogen, vitamins and
minerals, and a source of energy such as glucose (Mossel et al., 1995).

In the experiment, the media used were Nutrient Agar and Potato Dextrose Agar.
Nutrient Agar is a popular medium used to grow a variety of bacteria and fungi. The nutrients
included in this medium are peptone, beef and yeast extract, and sodium chloride. Its pH level is
neutral (7.4) at 25 degrees Celsius. On the other hand, PDA is mainly intended for the
cultivation of fungi and molds. It contains antibiotics that inhibit bacterial growth. Therefore, it is
a selective medium. It composition consists of potato infusion, dextrose, and agar. Its pH is 5.6
at 25 degrees Celsius. Half strength mediums are used in the experiment as some bacteria find
it too rich and cannot thrive in a high concentration. (Faddin, 1985).

The methods of bacterial transfer were air exposure and swabbing. In air exposure,
microbes in the air, if present, will stick to the surface of the agar if left for a period of time. In the
case of this experiment, 3 colonies were formed in the NA and 4 in the PDA when left in the
Multi-venture room for 2 minutes. Swabbing makes use of a sterile swab that is then swiped on
the desired surface. The nose area was swabbed and then transferred to an agar plate. After 24
hours, many colonies of bacteria were discovered.

Alcogel contains ethyl alcohol, a substance that kills bacteria through protein
denaturation and dissolving the lipid membrane. Alcogels are claimed to kill 99.9% of the
bacteria on our skin.In the experiment, the results show a alcogel to non-alcogel ratio of 40:10
and 1:2. Surprisingly, one agar plate shows that applying alcogel yielded more colonies of
bacteria while the other plate showed that alcogel reduced microbial count by 50%. Based on
the results, it can be said that the alcogel does not affirm its claim of reducing bacteria to 99.9%.
Possible source of error could be that the students have already applied alcohol prior to the
experiment. Another source of error is that contaminants from the air might have seeped into the
agar plate.

Conclusion
Based on the results, microorganisms can be found almost everywhere. Their growth
are affected by factors such as temperature, acidity, salt, nutrient content, and humidity. In
isolating microorganisms from the air, agar plate exposure can be used. On the other hand,
swabbing can be used for obtaining microbes on surfaces. Based on the results of our
experiment, alcogels do not kill 99.9% of the bacteria on the surface of the skin.
 References
Willey, J., Sherwood, L. & Woolverton, C. (2014). Prescotts Microbiology (9thed.). New York,
NY: The McGraw-Hill Companies, Inc.

Thiel, T. (1999). Introduction to Bacteria. University of Missouri

Mossel D., Corry J., Struijk C., & Baird R. (1995). Essentials of the microbiology of foods: a
textbook for advanced studiers. Chichester (England): John Wiley and Sons. 699 p.

Mac Faddin, J. F. 1985. Media for isolation-cultivation-identification-maintenance of medical

bacteria, vol.1. Williams & Wilkins, Baltimore, MD.