Kesini Casthuri Govindan Student ID: 3130300479

Microbiology Lab report

Experiment 3:

Distribution, disinfection and sterilization of bacteria

Abstract
The purpose of this experiment is to understand that the distribution of microorganism is
universal in nature including air, soil, water, animals, and human body. Most microbes do not
cause diseases and are not poisonous to human.

I. Microorganisms in air

Material Agar plate

Methods
1. Mark the bottom of one clean plate with name, date and 'air sample'.
2. Remove the lid of the agar plate and allow the plate to remain uncovered for the
entire lab period.
3. Replace the lid and incubate at 37ºC overnight. Observe the growing patterns of
microorganisms.

Result
Bacterial colonies grow on the plate.
Kesini Casthuri Govindan Student ID: 3130300479

II. Microorganisms on human skin

Material Agar plate

Method

1. Keep one's finger on the agar surface of a clean plate for a short time.
2. Incubate at 37ºC overnight. Observe the growing patterns of microorganism.

Result
Bacterial colonies grow on the plate.
Kesini Casthuri Govindan Student ID: 3130300479

III. Boiling and autoclaving

Principle
Heat is the simplest mean of sterilizing materials, provided the material is itself resistant to
heat damage. Heat acts by denaturing cell proteins and nucleic acids and by disturbing cell
membranes. A temperature of 100ºC will kill all vegetative forms but not spore forms of
bacteria within 2-3 minutes in laboratory scale cultures. A temperature of 121ºC for 15 min is
used to kill spores. There are two kinds of heat sterilization methods: dry heart and moist heat
(steam). At same temperature, steam is more effective because steam provides a means for
distribute heat to all parts of the sterilization vessel.

Materials
Broth culture of Bacillus subtilis and E.coli; broth tubes (12), pipette, autoclave, metal pot,
induction cooker.

Methods
1. Take 12 broth tubes, 2 for each bacteria, divide into three groups, mark the tubes with
group number and bacteria names (Bacillus subtilis or E.coli).
2. Using pipette, transfer two drops of bacteria (either Bacillus subtilis or E.coli) into
broth tubes according to labeling.
3. Treat the bacteria by boiling (group A) or autoclaving (group B). Leave one group as
untreated control (group C).
4. Incubate the broth tubes at 37ºC overnight. Observe the growing patterns of
microorganisms and record the result in the table below.
Kesini Casthuri Govindan Student ID: 3130300479

Results

Bacteria Group A Group B Group C

(100ºC, 5 min) (121ºC, 15 min) (no heating)
E.coli Clear: indicates no Clear Cloudiness
growth
Bacillus subtilis Cloudiness due to Clear Cloudiness
growth
Kesini Casthuri Govindan Student ID: 3130300479

IV. Ultraviolet radiation

Principle
The germicidal/ microbicidal effect of sunlight is due in large part to the action of ultraviolet
light. Microbicidal activity of ultraviolet (UV) light depends on the length of exposure which
is 20 mins ans the wavelength of UV which is ranged between 200nm-270 nm. Mechanism of
the microbicidal activity of ultraviolet (UV) light on microbes concerns the certain
photochemical effects on DNA. UV treatment leads to the forming of thymine-thymine
dimmers within the one DNA strand that interferes with base pairing and DNA replication.
Although UV light has a strong microbicidal effect, it has very low penetrating power; a
piece of paper can totally stop the UV lights penetrate. For this reason, UV light is commonly
used for air and surface cleaning.

Materials
Slant culture of E.coli agar plate, inoculation loop, alcohol burner, UV light source

Procedure
1. Streak the bacteria over the entire surface of an agar plate.
2. Take the plate to the UV light source and with the lids half-removed.
3. Expose it under the UV light for 20 mins.
4. Replace the lid and incubate at 37ºC overnight. Observe the growing patterns of
microorganisms.

Result
A large amount of bacteria colonies will grow on the area covered by the lid. No or rare
colonies will grow on the area directly exposed to UV light.