SANA MOUSAWI S4553520 HBM2105

LAB REPORT 1

SEPARATING MICROBES, ASEPSIS, HUMAN &

ENVIRONMENTAL MICROFLORA

By Sana Mousawi

INTRODUCTION/AIMS

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All the microorganisms that naturally inhabit our bodies or the

environment, including

bacteria, fungus, and viruses, are collectively referred to as the

microbiome. The human microbiome involves in metabolic processes,

battles against infections, trains the immune system, and via these

fundamental processes, impacts most of our physiological processes in

some way (Ogunrinola et al., 2020).

Microbes are everywhere, thus a laboratory has an abundant source of

possible contamination. The amount of contamination on tools and work

surfaces needs to be kept to a minimum to ensure the success of

experiments. Aseptic technique is a set of standard techniques used to

keep microbes out of sterilized fluids and cultures in the laboratory. These

techniques are necessary for experiments that involve cultivating cells.

The contamination possibility of fluids and cultures in an experiment is

decreased by practises like disinfecting laboratory workstation, making a

sterilized environment with a Bunsen burner, minimising the exposure of

open cultures to the air and sterilising the tools (Clare and Rowley, 2017).

The isolation streak method is a microbiological laboratory method for

obtaining isolated colonies of bacteria from a complex mixture or for

isolating pure bacteria cultures. It is commonly used to grow bacteria and

obtain pure cultures, but it may also be used to isolate yeasts. The

inoculum is streaked throughout the agar surface to possibly space

between individual bacterial cells (Eyler, 2013).

A microbial culture is a technique for growing microorganisms in a

controlled laboratory environment while allowing them to multiply in a

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predefined culture media. In microbiology, cultures are fundamental

diagnostic techniques that are employed as research tools (Eyler, 2013).

In real world, to better understand and treat infectious diseases, a pure

bacterial culture is necessary to research the bacteria's virulence,

sensitivity to antibiotics, and genomic sequencing (Choi et al., 2018).

The aim of this experiment is to learn and practice the aseptic technique

and the microorganism separating techniques. Also, examine the bacteria

in the air and on human body.

METHODS

As described in HBM2105, Medical Microbiology & Immunity, Laboratory

Manual 2, Semester 2 Block 2.

RESULTS

Exercise 1

1 4

Figure 1. The observed quadrant horse

blood
agar (HBA) streak plate, 37° C

2 3

Streak Shape/Co Size Texture Colour Number

Plate lony
37° C Round Small Moist/slim Yellow Hundreds

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Blood Circular y

Agar
Table 1. Specifications of the observed streak plate in Exercise 1

Exercise 2

Figure 2. The observed Nutrient

Agar (NA) plate, Air 37° C

Figure 3. The observed Nutrient Agar

plate, Air 25° C

Streak Shape/ Size Texture Colour Number

Plate Colony
NA Air Round/Circula Small Moist/slim Yellow & 5

37° C r y White
NA Air Round/Circula Small Moist/slim Yellow & 12

25° C r y White

Modera Brown 1

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Round/Circula te Dry Brown

Table 2. Specifications of the observed streak plate in Exercise 2

Exercise 3A,3B & 3C

Armpit

Figure 4. The observed trypticase-soy

agar (TSA) plate with swabs from
Individual 1’s armpit and in between
fingers, 30° C - Exercise 3A

Finger

Individual 1

Figure 5. The observed Baird Parker plate

(BPA) plate with fingerprints of two individuals, 30° C
– Exercise 3B

Individual 2

Individual 1

Figure 6. The observed mannitol

salt agar (MSA) plate with
5 fingerprints of two individuals, 30° C
– Exercise 3B
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Individual 2

Figure 7. The observed TSA plate

Afte Bef
with swab fromore the palm of Individual
r
1, 37° C – Exercise 3C

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Streak Shape/ Size Texture Colour Number

Plate Colony
TSA 30° Round/Circula Very Slimy/Moist Milky Thousands

C r small white

Armpit &

Finger
BPA 30° Punctiform Very Shiny Black ~200

C small

Individua

l1
BPA 30° Punctiform Very Shiny Black ~100

C Wavy/ small (Black)

Individua Filamentous Dry White 1

l2 Moderat

MSA 30° Lobate/Irregul Large Slimy/Moist White 4

C ar

Individua

l1
MSA 30° Round/ Small Slimy/ Moist Yellow ~60

C Circular &

Individua White

l2
TSA 37° Round/Circula Moderat Slimy/Moist White 10

C r e

Before

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TSA 37° Wavy Small Slimy/Moist Yellow Thousands

C Circular Moderat Dry White 1

After Circular e Moist/Transluc Opaque ~60

Small ent

Table 3. Specifications of the observed streak plates in Exercise 3A, 3B & 3C

DISSCUSION

The streak plate's primary goal is to create well-distributed bacterial

colonies from a concentrated cell solution. The bacteria were streaked

with an inoculating loop over solid media in several stages to get the

outcome. After each streaking step flaming was done to sterilise the loop

and more diluted inoculum. In microbiology, aseptic is important as it

maintains pure cultures and avoids contamination during transferring from

cultures to media. It is also important to wait until the sterilised loop is

fully cooled down to destroying the microorganisms (Clare and Rowley,

2017).

When the bacteria were being streaked from section 1 to 4 respectively,

dilution occurred. As the bacteria streaked in quadrant 1 was directly from

the culture, the bacterial growth was denser than the subsequent

quadrants. That is why according to Figure 1, the number of cells in

section 4 is minimal, which was anticipated. Each isolated cell replicated

and a visible pile of slimy, yellow and round colonies of bacteria formed

around the primary isolated cell.

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There are microorganisms in the air but the amount of them depends on

several conditions including temperature and humidity (Phillips, 2009).

According to Figure 2 & 3, the number of microorganisms in warmer air is

less than in cooler air, because it is harder for bacteria to survive in heat.

According to Table 2, both plates revealed same bacteria colonies, but the

“25° C plate” had more bacteria and a brown, dry bacteria.

Certain bacteria normally live on human’s skin. But the number of

bacteria in the wet areas of skin like the armpit, are proper spots for

bacteria to grow, as bacteria do better in the moist (Phillips, 2009). As

shown in Figure 4, there is equal or even somewhat more amounts of

bacteria in the “between fingers” sample section of the plate. This is

because the individual’s fingers have been slightly moist, as they were

wearing gloves before taking the sample.

According to Figure 5, in the “BPA plate”, Staphylococcus aureus with

black colonies and some with very narrow white margin appeared for both

Individuals 1 & 2. Staphylococcus aureus is a normal flora, which is found

on the human skin. According to Figure 6, Individual 1 fingerprints

revealed Staphylococcus epidermidis, which are the white colonies on the

upper part of the plate and “Individual 2” sample revealed Staphylococcus

aureus, which are the opaque and yellow colonies at the bottom of the

plate. Since everyone's skin has a distinctive fundamental structure, due

to inheritance, which enables interactions between various

microorganisms (Choi et al., 2018). The microbiome of Individual 1 & 2

from various locations varies because microbes also vary in the

community. Overall, the microbes on the human body and in the air or

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environment are different. Human skin contains microflora and there are

different microorganisms in the air possibly pathogens too. The

environmental microbiome effects the human microbiome and play

important role in human’s health. Hence why spitting has been socially

unacceptable in Australia, as it has been found to be associated with the

spread of infections (Ogunrinola et al., 2020).

From Exercise 3C, it is demonstrated that bacteria can be transferred by

touch easily.

According to Figure 7, the bacteria on the “Before” section is a few but

after the touch of Micrococcus luteus, a large colony appeared on the

“After” section of the plate.

Microorganisms play a vital role to life on earth. Therefore, understanding

bacteria and microbiology is crucial for comprehending the whole life on

earth. Ultimately, all the objectives of the experiment were achieved.

Knowledge of microorganism isolation principles, asepsis and microflora

were gained by the students.

REFERENCES

1. Choi, Q., Kim, H.J., Kim, J.W., Kwon, G.C. and Koo, S.H. (2018).
Manual versus automated streaking system in clinical microbiology
laboratory: Performance evaluation of Previ Isola for blood culture
and body fluid samples. Journal of Clinical Laboratory Analysis,
32(5), p.e22373. doi:10.1002/jcla.22373.

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2. Clare, S. and Rowley, S. (2017). Implementing the aseptic non touch

technique (ANTT) clinical practice framework for aseptic technique:
A pragmatic evaluation using a mixed methods approach in two
london hospitals. Journal of Infection Prevention, 19(1), pp.6–15.
doi:10.1177/1757177417720996.
3. Eyler, E. (2013). Chapter One - Pouring Agar Plates and Streaking or
Spreading to Isolate Individual Colonies. [online] ScienceDirect.
Available at:
https://www.sciencedirect.com/science/article/abs/pii/B97801242006
78000015.
4. Ogunrinola, G.A., Oyewale, J.O., Oshamika, O.O. and Olasehinde, G.I.
(2020). The Human Microbiome and Its Impacts on Health.
International Journal of Microbiology, 2020, pp.1–7.
doi:10.1155/2020/8045646.
5. Phillips, M.L. (2009). Gut Reaction: Environmental Effects on the
Human Microbiota. Environmental Health Perspectives, [online]
117(5), pp.A198–A205. Available at:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2685866/.
6. Sanders, E.R. (2012). Aseptic Laboratory Techniques: Plating
Methods. Journal of Visualized Experiments, [online] 3064(63).
doi:10.3791/3064.

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