MICROBIOLOGY

KBO20201P/YBO20101P

LAB REPORT 1: ASEPTIC TECHNIQUE

INDIVIDUAL HANDS-ON MARKS LAB REPORT

ID NUMBER MARKS TOTAL

STUDENT’S NAME MARKS

012023090028 AHMAD FAIZ BIN AHMAD FAUZI

012023071524 NUR HANNAH AJLAA BINTI

MUHAMAD NASARRUDDIN

012023091930 ROSHSEINI A/P MAKENTERAN

012023090111 TEO XIN LE

LECTURER: MS YASOHDHA ANNE

INTRODUCTION

Aseptic technique is a fundamental set of practices and procedures used in various

fields to create and maintain a sterile environment and to prevent contamination from

pathogens.

This practice should be used for procedures such as screening of isolates or strains,

pure cultures, slant cultures, single spore cultures and microbes transferring cultures.

Proper aseptic technique can prevent the cultures contamination from inborn and

outborn microbes in the environment and minimize the risk of contamination. It can also

avoid microbes from being unintentionally released into the environment and

contaminating the lab users in the laboratory.

Aseptic techniques are essential for producing accurate and reliable results, as

contamination can lead to false or misleading results that could impact research findings

and conclusions.

OBJECTIVES

To learn how to:

● decontaminate the lab bench

● organize the workspace

● adjust the bunsen burner

● Sterilize inoculating tools

● Aseptically transfer organisms from broth/plate cultures using BSL 2 procedures

MATERIALS AND APPARATUS

A) Aseptic Technique

1) Bunsen burner

2) Microbial culture

3) 70% alcohol

4) Sterile plate with nutrient agar

5) Inoculating loop

6) Match/lighter

7) Tissue paper (for wiping the bench)

8) Disposable gloves

B) Bacteria culture

1) Nutrient agar plates

2) Sterile cotton swab

3) Bacteria sample

4) Incubator
PROCEDURES

A) Flaming procedure

The flaming procedure is designed to heat the end of the loop gradually because after

use it will contain culture, which may “splutter” on rapid heating with the possibility of

releasing small particles of culture and aerosol formation.

1. The handle was positioned at the end of the wire in the light blue cone of the

flame. This was the cool area of the flame.

2. The rest of the wire was drawn upwards slowly up into the hottest region of the

flame, immediately above the light blue cone.

3. It was held until it was red hot.

4. The full length of the wire was ensured to receive adequate heating.

5. It was allowed to cool then used immediately.

6. The loop did not put down or it was waved

around.

7. The loop was resterilized immediately

after use.
B) Flaming the neck of bottles and test tubes

1. The lab table was thoroughly disinfected by applying 70% ethanol and it was

allowed to air dry.

2. An inoculating loop was obtained and a culture tube containing a microorganism.

3. The Bunsen burner was lighted and a blue cone flame was adjusted to obtain

(Figure 1).

4. The culture tube cap was loose but it can't be removed.

5. The lid of the bottle was loose so that it could be removed easily.

6. The bottle/test tube was lifted with the left hand.

7. The lid of the bottle/cotton wool plug was removed with the little finger of the right

hand. (The bottle was turned, not the lid.)

8. The lid/cotton wool plug didn't put down.

9. The neck of the bottle/test tube was flamed by passing the neck forwards and

back through a hot Bunsen flame (see Figure 2).

10. The lid on the bottle/cotton wool plug was replaced using the little finger (see

Figure 2). (The bottle was turned, not the lid.)

11. The lid of the nutrient agar plate was lifted, as little as possible, and the bacteria

was spreaded on the inoculating loop over the surface of the agar (see Figure 3).

The Petri dish was rotated 90° and repeated.

12. The inoculating loop was reflamed until red hot (see Figure 1).

13. The lab table was thoroughly disinfected as directed in step 1.

C) Swabbing procedure

1. The lab bench was thoroughly disinfected by applying 70% ethanol and it was

allowed to air dry.

2. A sterile cotton swab was used to swipe the surface of the phone twice.

3. Once the cotton swab was coated with the bacteria, it was streaked in a zig-zag

pattern on agar plate A.

4. The fingerprint of the thumb was placed on agar plate B.

5. A new sterile cotton swab was used to rub the sweaty area on the body. The

bacteria collected with the swab was then transferred on agar plate C.

6. The three agar plates were then kept in an incubator.

7. The observation was recorded.

RESULTS

Agar plate Diagram Observations

Ros and Little to few bacteria colonies

Anabelle’s present on both sides of the
phone swab agar plate.

Hannah and A higher density of bacteria

Faiz’s phone colonies spread evenly across
swab the agar plate.
Thumbprint of Anabelle and Hannah’s
Ros, Anabelle, fingerprints had a few bacteria
Hannah and colonies present.
Faiz
While Ros and Faiz’s
fingerprint had a higher density
of bacteria present.

Upper lip swab An abundance of bacteria

of Faiz colonie was present on the
agar plate.
DISCUSSION QUESTIONS

1) Why is aseptic technique important in the diagnosis of a disease?

- This is to prevent microbes from unintentionally being released into the

environment and contaminating the laboratory and users.

2) Why do we use 70% alcohol as part of aseptic technique?

- As a disinfectant, 70% alcohol is the balanced concentration for killing bacteria

and viruses. At 70% concentration, even though ethanol takes longer to

evaporate, it is able to penetrate cells more effectively. Additionally, the water in a

70% grade is crucial in denaturing proteins. It is also less flammable.

CONCLUSION

In conclusion, aseptic technique is important to create and maintain a sterile

environment and prevent contamination by unwanted microorganisms. Whether working

with microorganisms in a laboratory or performing medical procedures, aseptic

technique plays a critical role in safeguarding human health, ensuring accurate research

and experimentation, and maintaining product quality in various industries.

REFERENCES

● Kaiser, G. (2021, May 5). 2.2: Introduction to Bacterial Growth and Aseptic
Techniques. Biology LibreTexts.
https://bio.libretexts.org/Courses/North_Carolina_State_University/MB352_Gener
al_Microbiology_Laboratory_2021_(Lee)/02%3A_Cultivation_of_Microbes/2.02%
3A_Introduction_to_Bacterial_Growth_and_Aseptic_Techniques
● Reynolds, J. (2016, March 26). Bacterial Colony Morphology. Biology LibreTexts.
https://bio.libretexts.org/Learning_Objects/Laboratory_Experiments/Microbiology
_Labs/Microbiology_Labs_I/08%3A_Bacterial_Colony_Morphology

● Microbiology Society. (2020). Observing bacteria in a petri dish. Microbiology

Society.
https://microbiologysociety.org/why-microbiology-matters/what-is-microbiology/ba
cteria/observing-bacteria-in-a-petri-dish.html

● Gelling, C. (2023). Ethanol Grades: The 3 Types Of Ethanol Used in the Lab.
Bitesize Bio.
https://bitesizebio.com/13518/which-type-of-ethanol-should-i-use/#:~:text=At%20
a%2070%25%20concentration%2C%20however,is%20less%20flammable%20%
5B1%5D.
● Cherney, K. (2018, September 29). Aseptic technique. Healthline.
https://www.healthline.com/health/aseptic-technique#types

● Siddiquee, S. (2017). The basic concept of microbiology. In Fungal biology (pp.

1–15). https://doi.org/10.1007/978-3-319-64946-7_1
RUBRIC