KOLEHIYO NG LUNGSOD NG DASMARIÑAS

Laboratory Manual
General Microbiology

P
repared by:

nst. Lou Sandino L. Castro

 Institute of Applied Sciences
Science Unit

EXPERIMENT 2 Hay Infusion

OBJECTIVES

▪ Understand the diverse world of microorganisms, their roles in ecosystems, and their
significance in nutrient cycling and decomposition.

▪ Develop skills in observing and identifying microorganisms using the microscope.

▪ Identify the microorganisms observed in the hay infusion and their locomotory
organelles.

▪ Collaborate with peers in preparing the hay infusion, sharing observations, and
discussing findings to enhance collective learning and understanding.

DISCUSSION

A hay infusion is a mixture made by soaking hay or dried grass in water. This mixture
serves as a nutrient source for microorganisms, which naturally exist on the surfaces of
plants and in the environment. When hay is placed in water and allowed to soak, it releases
organic materials and nutrients into the water, creating an environment conducive to the
growth of various microorganisms. The resulting mixture becomes a microcosm, a small-
scale representation of a natural ecosystem. It enables scientists and students to observe
the dynamics of microbial communities, nutrient cycling, and the interactions between
different microorganisms in a controlled setting.

In this activity, you can expect to observe a variety of microorganisms thriving within the
mixture. The types of microorganisms present can vary depending on the environment
from which you collected your samples and the duration of the experiment. Here are some
common microorganisms you might observe in a hay infusion:

1. Bacteria: Bacteria are ubiquitous microorganisms that come in various shapes,

sizes, and arrangements. They play essential roles in nutrient cycling, decomposition, and
other ecological processes.
2. Algae: Algae are photosynthetic microorganisms that can range from single-celled
to multicellular forms. They may appear green, brown, or even red-colored cells,
depending on the species.
3. Protozoa: Protozoa are single-celled organisms that move using hair-like structures
called cilia or whip-like structures called flagella. They are often predators, feeding on
bacteria and other smaller microorganisms.

NS0300L- General Microbiology

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 Institute of Applied Sciences
Science Unit

4. Rotifers: Rotifers are microscopic multicellular organisms with cilia surrounding

their mouths, creating water currents that bring in food particles. They are filter feeders
and can be seen rotating as they move.
5. Euglenoids: Euglenoids are single-celled organisms that can be both autotrophic
(photosynthetic) and heterotrophic (feeding on other organisms). They have a whip-like
tail called a flagellum.
6. Amoebas: Amoebas are unicellular organisms that move by extending and
retracting their pseudopodia (temporary “feet”). They feed by engulfing smaller particles.
7. Paramecia: Paramecia are ciliate protozoa with an elongated body covered in cilia.
They have a characteristic slipper-like shape and are voracious feeders.

MATERIALS

Hay Infusion Preparation

Qty Specs Item

500 mL Clean Water
3-5 mL Canal or Pond water
1 Glass jar
10 grams Hay or dried grass (handful)
1 Scissors
1 Wire loop
1 Alcohol lamp
1 Tripod
1 Lighter
100 mL Beaker
50 mL Beaker
1 Stirring rod

Observation in the Laboratory

Qty Specs Item

1 Light microscope
1 Glass slide
1 Cover slip
1 dropper

SAFETY PRECAUTIONS

NS0300L- General Microbiology

PAG
E \*
 Institute of Applied Sciences
Science Unit

● Observe proper handling of the microscope.

● Handle microscope slides and coverslips carefully to avoid cuts from broken glass.

● Do not play with the heat and keep lighter away when not in use.

● Adhere to all laboratory rules and guidelines.

PROCEDURE

Hay Infusion Preparation

1. Boil 1 glass of clean water.

2. Add hay or dried grass and simmer for 3 minutes.
3. Transfer the preparation into a clean glass jar and allow to cool.
4. Add 2-3 tablespoons of canal or pond water. Cover the glass jar half Way and store at
room temperature for 2-3 days.

Observation in the Laboratory

1. Using a dropper, draw a sample from the bottom of the glass jar and place 1 drop on
a clean glass slide and place the coverslip.
2. Observe under the microscope.
3. Record at least 3 microorganisms observed in the hay infusion.

DISPOSAL AND CLEAN-UP

● All equipment and materials must be clean and dry properly.

● Microscope must be returned to its proper storage.
● Working table must be cleaned.
● Dispose of used microscope slides, coverslips, and other waste materials properly
according to your lab's disposal protocols.
REFERENCES

NS0300L- General Microbiology

PAG
E \*
 Institute of Applied Sciences
Science Unit

NAME: ____________________________ RATING: ____________

LAB REPORT FOR
EXPERIMENT 1 GROUP: _________

DATE SUBMITTED: ___________________

Results and Observations

Fill in the box below by providing a photo documentation of the different specimens you recorded, label their
parts (nuclei, cell walls or cell membranes, and locomotory organelles, and briefly explain their role in the
ecosystem. (10pts)

Observed specimen 1 Observed specimen 2

Species name: Species name:

_______________________________ _______________________________

Description: Description:
____________________________________ ____________________________________
____________________________________ ____________________________________
____________________________________ ____________________________________
____________________________________ ____________________________________
____________________________________ ____________________________________
NS0300L- General Microbiology
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 Institute of Applied Sciences
Science Unit

Observed specimen 3 Observed specimen 4

Species name: Species name:

_______________________________ _______________________________

Description: Description:
____________________________________ ____________________________________
____________________________________ ____________________________________
____________________________________ ____________________________________
____________________________________ ____________________________________
____________________________________ ____________________________________

Post-Lab Questions

1. What were the types of microorganisms you observed in the hay infusion mixture? Enumerate
them and describe their behavior. (5 pts)

NS0300L- General Microbiology

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 Institute of Applied Sciences
Science Unit

2. What factors affect the growth of microorganisms in a hay infusion mixture? (5pts)

3. What is the importance of boiling hay or dried grass in the preparation of hay infusion? (5 pts)

4. What locomotory organelles do euglenas and paramecia have? Describe their motility. (5 pts)\

5. What is the importance of adding canal or pond water in the hay infusion preparation? (3pts)

6. What are the differences between prokaryotic and eukaryotic organisms? (3pts)

NS0300L- General Microbiology

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E \*
 Institute of Applied Sciences
Science Unit

7. Are the protozoans in the hay infusion pathogenic to humans? Support your answer with
evidence.

8. What is the difference between autotrophic and heterotrophic organisms?

REFERENCES:

Summary and Conclusions

NS0300L- General Microbiology

PAG
E \*
 Institute of Applied Sciences
Science Unit

NS0300L- General Microbiology

PAG
E \*