BIOLOGY 3

MICROBIAL DIVERSITY
LABORATORY REPORT NO. 1
Hay Infusion

prepared by PAUL EDUARD A. SAGARINO

A. Method Summary (in your own words and in paragraph form)

I. Preparation of Hay Infusion

The class prepared a hay infusion mixture for everyone. We heated the leaves
on a hot plate at 345 C and allowed it to boil for six hours. Afterwards, a classmate
drained some of the water and removed the majority of the pond leaves. She then
added pond water into the beaker and it was placed on a rack to rest under ambient
light.

II. Microscopic Examination

The class borrowed depression slides and cover slips. I dropped 2 drops of water
from the beaker with hay infusion water and I cut a small piece of leaf from the beaker. I
placed a small piece of cotton to slow down the organisms in the drops of hay infusion
water. I placed the depression slide on the microscope and looked at the organisms and
identified them with the help of our adviser, sir Millard.

B. Observations / Results (photographs with captions, text, paragraphs, etc.)

I. Preparation of Hay Infusion

Figure 1.1 Hay Infusion after boiling

 Figure 1.2 Hay Infusion

II. Microscopic Examination

Figure 2.1 Colpidium under a microscope

Figure 2.1 shows a Colpidium from the hay infusion under a microscope.
 Figure 2.2 Binary fission and euglena under a microscope

Figure 2.2 shows a binary fission and euglena from the hay infusion under a
microscope.

Figure 2.3 Paramecium under a microscope

Figure 2.3 shows a Paramecium from the hay infusion under a microscope.
 Figure 2.4 Amoeba under a microscope

Figure 2.4 shows an Amoeba from the hay infusion under a microscope.

1) Ciliates, flagellates, and amoebae exhibit unique forms of movement when observed
under the microscope. Ciliates move rapidly and smoothly using coordinated hair-like
cilia, sometimes appearing erratic as they change direction quickly. Flagellates, on the
other hand, propel themselves through their environment using whip-like flagella, often
swimming in a spiral or helical pattern. Amoebae employ a slower, crawling motion
called amoeboid movement, which involves the continuous extension and retraction of
temporary cellular projections called pseudopods, resulting in a constantly changing
shape. The diverse locomotion strategies of these microscopic organisms demonstrate
the remarkable adaptability of life at the cellular level.

C. Discussion (explain what you saw / explain your results)

I. Preparation of Hay Infusion

Hay infusions are boiled to sterilize them and kill any bacteria or microorganisms
that may be present in the hay. They are then placed on a rack with ambient lighting to
provide exposure to oxygen and light, which are necessary for the growth of
microorganisms. The smell of hay infusions can be unpleasant due to the
decomposition of organic matter in the hay, which produces gasses like methane and
hydrogen sulfide. However, this is a natural part of the process of creating a diverse
microbial community in the hay infusion.

We added leaves to the hay infusion to increase the diversity of microorganisms

present in the infusion. Different types of leaves contain different nutrients and organic
matter that can support the growth of a wider range of microorganisms. This can lead to
a more diverse microbial community in the hay infusion, which can be useful for a
variety of purposes such as studying microbial ecology or testing the efficacy of
antimicrobial agents. Additionally, adding leaves can help to mimic the natural
environment in which the hay and other plant materials would be found, which can
enhance the infusion.
 II. Microscopic Examination

In the early stages of a hay infusion, Paramecium may be the dominant

microorganism due to its rapid growth, ability to exploit abundant resources, and
tolerance to fluctuating environmental conditions. As the hay infusion matures over a
week or two, conditions stabilize, and resources become more limited, allowing
slower-growing organisms like amoebas to appear. The shift in environmental
conditions, competition for resources, and predation contribute to the development of a
more diverse microbial community. Paramecium's initial advantage in a nutrient-rich
environment gives way to the establishment of other protozoa, such as amoebas, which
can better compete as conditions change. Over time, the hay infusion ecosystem
becomes increasingly complex, reflecting the dynamic balance of species and their
adaptations to the environment.

The purpose of placing a small strand or piece of cotton fiber in the depression
slide with the drops of water from the hay infusion liquid is to impede the rapid
movement of certain microorganisms, such as ciliates and flagellates. By slowing down
these fast-moving organisms, it becomes easier to observe and study their morphology
and behavior under the microscope. The cotton fiber acts as a physical obstacle that
restricts their free movement, facilitating a more in-depth examination.

Ciliates are a group of single-celled protozoa characterized by the presence of

hair-like structures called cilia on their surface. These cilia are primarily used for
locomotion and, in some cases, for feeding. Paramecium is a well-known example of a
ciliate and has a characteristic slipper-like shape.

Paramecium swims by coordinating the beating of its cilia, which cover its entire
cell surface. The cilia beat in a synchronized manner, generating wave-like motions that
propel the organism through the surrounding medium. Paramecium can move rapidly
and change direction quickly, making it an efficient swimmer.

Feeding in Paramecium involves the use of specialized cilia-lined oral grooves,

also known as the cytostome. As the cilia around the oral groove beat, they create
water currents that draw in food particles, such as bacteria and other small
microorganisms. The food particles are then trapped in a coating of mucus and funneled
into the cell through the cytostome.

Flagellates are a group of single-celled protozoa that use whip-like appendages

called flagella for locomotion and, in some cases, for feeding. Euglena is a well-known
example of a flagellate, characterized by its elongated shape and the presence of a
single flagellum.

Euglena swim by rotating their flagellum, which extends from the anterior end of
the cell. The flagellum acts like a propeller, generating a force that pushes the organism
through the surrounding medium. As the flagellum rotates, Euglena often moves in a
spiral or helical pattern, which can be easily observed under a microscope.
 Euglena have a unique mode of nutrition called mixotrophy, meaning they can
obtain nutrients through both photosynthesis and phagotrophy. They possess
chloroplasts, which enable them to perform photosynthesis, converting sunlight into
chemical energy. This allows Euglena to produce their own food when light is available.

Amoebas are a group of single-celled protozoa that exhibit a distinctive mode of

movement and feeding mechanism. They are characterized by their irregular shape and
the ability to form temporary cellular projections called pseudopods.

Amoebas do not "swim" in the traditional sense. Instead, they move through a
process called amoeboid movement. This involves the extension and retraction of
pseudopods, which allows them to crawl or glide through their environment. Amoebas
form pseudopods in the direction they want to move, and their cytoplasm flows into the
projection. The rear part of the cell then retracts, propelling the organism forward. This
movement is generally slower than that of ciliates or flagellates and is characterized by
a continuous change in the organism's shape.

Amoebas feed through a process called phagocytosis, which also involves the
use of pseudopods. When an amoeba encounters a food particle, such as a bacterium
or another protozoan, it extends its pseudopods around the prey, eventually engulfing it.
The engulfed particle becomes enclosed in a food vacuole within the cell. The food
vacuole then fuses with lysosomes, which contain digestive enzymes that break down
the food particle into nutrients. These nutrients are absorbed by the cell, while
indigestible waste is expelled through the cell membrane via a process called
exocytosis.