MAPUA UNIVERSITY

SCHOOL OF CIVIL ENGINEERING & ENVIRONMENTAL & SANITARY ENGINEERING

FLORDELIZA C. VILLASENOR

___________________________________________________________________

MICROBIOLOGY and PARASITOLOGY

for ENVIRONMENTAL and SANITARY
ENGINEERING
_______________________________________

Compiled by:
FLORDELIZA C. VILLASENOR

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MAPUA UNIVERSITY
SCHOOL OF CIVIL ENGINEERING & ENVIRONMENTAL & SANITARY ENGINEERING
FLORDELIZA C. VILLASENOR

MICROBIOLOGY and PARASITOLOGY

LABORATORY

NAME: __________________________________________________
COURSE & SECTION: ______________ STUDENT NO.:________
GROUP NO. _____________

EXPERIMENT NO. 1
MICROORGANISMS in the ENVIRONMENT
TITLE

DATE PERFORMED: ___________________________

DATE SUBMITTED : ___________________________ GRADE

INSTRUCTOR

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MAPUA UNIVERSITY
SCHOOL OF CIVIL ENGINEERING & ENVIRONMENTAL & SANITARY ENGINEERING
FLORDELIZA C. VILLASENOR

EXPERIMENT NO. 1
MICROORGANISMS in the ENVIRONMENT
Theory
Microorganisms play some very important roles in the environment and in human health. A
small number of these microorganisms can cause diseases in humans which can be transmitted
through air, water, soil, food, insects and other paths. However, some microorganisms can
beneficial to human activities like treatment of sewage and biodegradable solid wastes. Thus,
recognizing the presence of certain kinds of microorganisms could help people in control and
prevention of diseases as well as improving the quality of life of humans.
It is important to note that microorganisms exist in mixed populations. Pure cultures of bacteria
rarely exist outside the research laboratory. Colonies can be grown from by culturing
microorganisms from different sources into agar plates. By getting samples from the
environment and putting samples in appropriate substrate, the microorganisms can grow into
colonies, thus validating their presence or absence in specific environmental compartments.

Outcomes
At the end of the experiment, the students should be able to identify the characteristics of the
microorganisms present in the environment.

Objectives
At the end of the experiment, the student should
• Be able to know the process of isolating microorganisms from the environment using
aseptic technique of culturing bacteria
• Be able to describe the forms oor shapes of colonies in agar plates
• Be able to identify the presence of microorganisms from different sources

Equipments
• 10 pcs. Of Petri dishes
• 2 liters of distilled water
• Sterile cotton swabs
• Permanent marker
• Safety gloves

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MAPUA UNIVERSITY
SCHOOL OF CIVIL ENGINEERING & ENVIRONMENTAL & SANITARY ENGINEERING
FLORDELIZA C. VILLASENOR
• Nutrient agar
• Autoclave
• 1-lter Erlenmeyer flask
• Bunsen burner
• Inoculating loop
• Reagents required for the determination of D.O.

Procedures
A. Preparing sterile agar plates.
1. Sterilize 1-liter Erlenmeyer flask, stirring rods, petri dishes (and covers) into an autoclave
Set at 121℃, 15 psi for 15 minutes.
2. After autoclaving, cool the petri dishes at room temperature with covers placed.
B. Preparing the agar plate
1. Dissolve nutrient agar in 1 liter of distilled water. Follow the manufacturer’s instruction
indicated in the label of the nutrient agar container.
2. Place the Erlenmeyer flask on electric stove continuously stirring the solution until it
boils.
3. Remove the flask from the stove.
4. Pour the nutrient agar solution into each Petri dish up to half-full. (Partially remove the
cover while pouring then replace the cover immediately.)
5. Leave the Petri dish for about 30 minutes until the solution congeals.
C. Inoculating the agar plate
Inoculates the agar plates with the following materials

Materials Method of Procedure

Inoculation
Food plate Inoculating Loop 1. Sterilize inoculating loop by passing
it over flame of a Bunsen burner until
the loop turn to flaming red. Let it
cool down for few seconds.
2. Partly remove the cover of the Petri
dish. Using the loop, get sample from
food plates and spread the material
over the agar plate. Return the cover
immediately.
Water Pour Pour few drops of water into the agar
plate while it is slightly open. Replace
cover immediately.

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MAPUA UNIVERSITY
SCHOOL OF CIVIL ENGINEERING & ENVIRONMENTAL & SANITARY ENGINEERING
FLORDELIZA C. VILLASENOR
Air Exposing to air Remove cover of the Petri dish. Leave
the agar plate exposed to air for at least
15 minutes. Replace the cover
immediately.
Garden Soil Sprinkling Sprinkle sparingly the garden soil on the
agar plate.
Sneezing, Direct inoculation Remove the cover of the Petri dish.
Coughing, Hair Cough, sneeze or smear the agar plate
Scalp with fingers
Surface (door knob, Swabbing Using sterile cotton buds, swab the
desk top) surface using cotton buds. Smear into
the surface of the agar plate. Replace the
cover immediately.
Human surface Swabbing Using sterile cotton buds, swab the
(skin, tow webs, surface using cotton buds. Smear into
molar) the surface of the agar plate. Replace the
cover immediately.

1. Set the temperature of incubator to 37℃. Put the agar plates inside the incubator
with each plate placed upside down. Leave the agar plates in the incubator for 7
days.
2. After seven days, examine each plate. Describe the kinds of colonies formed in
each plate using standard forms and elevation descriptions. Count the number that
each colony formed

Guide Questions
1. What are microorganisms?

2. What are the classifications and structures of a microorganism?

3. What are the importance of a microorganisms to us people?

4. Discuss the evolution of the microorganisms through time.

5. Why do we need to study microorganisms?

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MAPUA UNIVERSITY
SCHOOL OF CIVIL ENGINEERING & ENVIRONMENTAL & SANITARY ENGINEERING
FLORDELIZA C. VILLASENOR

Data Sheet
EXPERIMENT No. 1:
MICROORGANISMS in the Environment
Course/Section: __________________________Date Performed: ________________________
Members:
Name Signature
1. __________________________________________ ________________________

2. __________________________________________ ________________________
3. __________________________________________ ________________________
4. __________________________________________ ________________________

Observe each agar plate and record the observations based on form, elevation, margin and number
of colonies.

___________________________________________________________________
Data:

1. __________________________________ 2. ___________________________________

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MAPUA UNIVERSITY
SCHOOL OF CIVIL ENGINEERING & ENVIRONMENTAL & SANITARY ENGINEERING
FLORDELIZA C. VILLASENOR

Form Elevation Margin No. Form Elevation Margin No.

3. __________________________________ 4. ___________________________________

Form Elevation Margin No. Form Elevation Margin No.

Instructor’s Signature
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MAPUA UNIVERSITY
SCHOOL OF CIVIL ENGINEERING & ENVIRONMENTAL & SANITARY ENGINEERING
FLORDELIZA C. VILLASENOR

5. __________________________________ 6. ___________________________________

Form Elevation Margin No. Form Elevation Margin No.

7. __________________________________ 8. ___________________________________

Instructor’s Signature

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MAPUA UNIVERSITY
SCHOOL OF CIVIL ENGINEERING & ENVIRONMENTAL & SANITARY ENGINEERING
FLORDELIZA C. VILLASENOR

Form Elevation Margin No. Form Elevation Margin No.

9. __________________________________ 10. ___________________________________

Form Elevation Margin No. Form Elevation Margin No.

Instructor’s Signature

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MAPUA UNIVERSITY
SCHOOL OF CIVIL ENGINEERING & ENVIRONMENTAL & SANITARY ENGINEERING
FLORDELIZA C. VILLASENOR

Analysis

Conclusion

Reflection

References

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