Experiment 1

THE ANIMAL-LIKE PARAMECIUM

Carandang, Cruz, Pasumbal, Salem, Tolentino

ABSTRACT

This experiment deals with the observation and study of Paramecium. Cultures were observed
under the microscope to observe its gross morphology, feeding habits and reaction to various stimuli.
Physical obstructions were placed to observe thigmotaxis, exposure to various chemicals was done to test
for chemotaxis. Cultures were placed in test tubes and partially exposed to light to determine specien
reaction to light and gravity. Electrical currents at increasing increments were applied to specimens under
the microscope to observe galvanotaxis. Contractile vacuole activity was monitored by observing
specimens placed under solutions of varying saline levels. Digestion was observed under the microscope
through the use of a dyed yeast suspension. Experimental data obtained show the decline of contractile
vacuole activity as the salinity of the media increases, and is in compliance with results found in
literature. Chemotaxis tests showed that specimens reacted positively to sodium bicarbonate and sodium
chloride which is in contrast to theoretical results; it also reacted positively to sucrose. The response of
Paramecium to gravity differed from the expected result - the specimen aggregated near the bottom of the
container. The specimen was also observed to be positively phototactic. Galvanotaxis experiments
demonstrated an affinity of specimen to swim towards the cathode - the speed varying with voltage.
However, cell death occurred on the highest voltage.
INTRODUCTION negative taxis shows that the Paramecium
avoids the stimuli introduced [REF]
Paramecium are unicellular ciliates that belongs
to Phylum Ciliophora. The structures present in In this experiment, the researcher observed the
the unicellular eukaryote allows it to interact and (1) morphology of the Paramecium (2)
detect the changes in its environment. They are movement (3) taxis and (4) the role of its
considered as animal-like organisms due to their contractile vacuole in different levels of salinity
ability to move, reproduce, feed and react to of its habitat.
different taxis. The taxis which varies from
light, gravity, touch, salinity and chemicals all MATERIALS AND METHODS
contribute to the tendency and behavior of the
Paramecium when it is introduced to the To observe gross morphology and movement,
medium it inhabits. [REF] one drop of a Paramecium culture was mounted
on a slide along with a thin sheet of wet cotton
The reaction of the Paramecium can be to entrap Paramecium, under this setup the
observed by preparing a drop of the sample Paramecium’s movement in response to
culture in a glass slide under the microscope. physical obstructions (thigmotaxis) were
The reaction of the Paramecium to the factors observed.
may be positive or negative depending on its
effects to the chemosensory organ of the Paramecium reaction to chemical stimuli
Paramecium. A positive taxis indicates that the (chemotaxis), was observed by exposing
Paramecium is drawn to the stimuli while a specimens to 0.1 N HCl, 0.2% acetic acid, 0.1 M
Sodium Bicarbonate, 3% NaCl, and 5% sucrose
solutions. Pieces of threads are coated in each of
the solutions and placed on a slide containing
the culture then mounted under a microscope.
Geotaxis and phototaxis were observed observe
through placing Paramecium cultures into 2 test
tubes, one covered partly with aluminum foil,
storing them vertically and exposing them to
light. Visual observation was performed to
determine the effects of light and gravity on the
specimen.
a transparent circular organelle present in the
anterior and posterior part of the sample. The
food vacuole, as shown in figure 2 can be
observed when the Paramecium sample ingested
food stained with congo red. The digestion of
food vacuole stained by congo red was observed
to be red in color. After digestion, the expelled
particles were observed to be dark red in color.

Galvanotaxis experiments were performed by

construction of a circuit and exposing specimen
to the electrical current. The voltage used was
increased incrementally to observe the nature of
its effect.

The role of the contractile vacuole was observed

by manually counting the number of their
Figure 1. Morphology of Paramecium in 5%
pulsations under different conditions provided
NaCl solution under high power objective and
by differing media. Paramecium were observed
4x zoom showing the cilia and macronucleus.
in its culture media, distilled water, 2.5%, 5%,
7.5% and 10% NaCl solutions and KCN.
Acidity of the media was determined by the use
of neutral red.

To observe the digestion of Paramecium, 0.1g

of yeast were boiled in 40 mL of water. Congo
red dye was added to the yeast solution to
visualize the passage of food within the
specimen. One drop of the dyed suspension was
added to one drop of the culture and feeding was
observed under the microscope.

RESULTS Figure 2. Food vacuole containing yeast stained

The gross morphology of the Paramecium was
observed under the high power objective of a
light microscope. As shown in figure 1, the cilia,
which is responsible for locomotion and food
capture covers the pellicle of the organism. The
contractile vacuole, which allows the function of
osmoregulation inside the cell is also present as
with Congo Red.
The role of the contractile vacuole in
osmoregulation is observed by subjecting the
ciliates to different concentrations of NaCl.
Shown by table 1 below, the frequency of
pulsation of the contractile vacuole generally
 decreases and the concentration of NaCl sodium bicarbonate as indicated by a (+).
increases. Similar results were also obtained in the 3%
NaCl and 5% sucrose indicating that it is
Table 1. Effect of NaCl concentration on the attracted to these reagents.
pulsation of the contractile vacuole.
Solution Number of contractions Table 3. Chemotaxis reaction of Paramecium to
per minute different reagents
Distilled water 32 Reagents Chemotaxis (+/-)
Culture medium 12 1 M HCl -
2.5% NaCl solution * 0.2% Acetic Acid -
5% NaCl solution * 0.1 M sodium +
7.5% NaCl solution 5 bicarbonate
10% NaCl solution 2 3% NaCl +
* Number of contractions cannot be observed, the 5% sucrose +
Paramecium cells present in the culture were
inactive. Paramecium showed positive geotaxis and
phototaxis. The Paramecium cells aggregated on
To observe the different reaction of the bottom of the container which is exposed to
Paramecium to different stimuli, the stimuli of light.
touch, reagents, light, gravity and electric
current in the sample were introduced. The The tendency of Paramecium to either approach
reaction of the Paramecium to touch was or avoid electric currents were tested by
induced by using a cotton fiber which shows the subjecting the culture medium increasing
tendency of Paramecium to avoid any obstacle. amount of voltage from 1.5 V, with a partition of
If the anterior side is obstructed, it will avoid 1.5, to 12 V by using a D type battery. As seen
this by going backward and changing directions in Table 3, the voltage from 1.5 to 9.0 volts gave
while a hit in the posterior part will just be a galvanotaxis of (+) indicating that the
ignored by the organism. Paramecium migrated towards the cathode of
the battery. The result for the 10.5 and 12 V
Table 2. Thigmotaxis reaction of Paramecium to however showed that the Paramecium decreased
cotton strand its rotation or died because of the current flow.
Anterior part Move backwards and change Further increase in the voltage beyond 12 V was
direction not observed due to the samples cannot tolerate
Posterior part Continue to move forward the voltage anymore.

The chemotaxis was observed by introduction of

Table 4. Galvanotaxis reaction of Paramecium
1 M HCl, 0.2% acetic acid, 0.1 M sodium
to different voltages of electricity.
bicarbonate, 3% NaCl and 5% sucrose. As
Voltage Galvanotaxis
shown in table 2, the results in 1 M HCl
1.5 (+)
,indicated by a (-) sign, showed that the 3.0 (+)
Paramecium avoided the strong acid and some 4.5 (+)
of the organisms which came in close proximity 6.0 (+)
also died. Similarly, the acetic acid, which is a 7.5 (+)
weak acid, also shows similar results. The 9.0 (+)
Paramecium was attracted however to 0.1 M 10.5 Slow rotation
 12.0 death
DISCUSSION
Gross Morphology and Movement
Figure 3. A diagram showing the important
organelles and cell structures of Paramecium in
digestion and osmoregulation. (Adapted from
Urry et al, 2014).
The morphology of Paramecium consists of a
thin, blunt anterior end, and a thick, sharper
posterior end. It has a micronucleus, which
functions for conjugation, and a macronucleus,
which functions for metabolism. Its two
contractile vacuoles, surrounded by radiating
canals, functions for osmoregulation by taking
in excess water then pumping it out. During
digestion, it has an oral groove, a vestibule, a
cytosome, a cytopharynx and an anal pore
through which food passes to these structures
respectively. [REF]
The Paramecium moves through its ciliary
movement. The cilium performs whiplash
movements to propel water in which has two
phases, the effective stroke and the recovery
stroke. The fast effective stroke happens when
the cilium moves in the direction of the main
flow of fluid while the recovery stroke occurs
when the cilium moves back close to the cell
surface to prepare for the next effective stroke.
Usually, the recovery stroke is slower than the
effective stroke because the cilium curls to one
side and sweeps in counter-clockwise. (Blake &
Sleigh, 1974).
Figure 4. Effective stroke (above), recovery
stroke (below). (Adapted from Blake & Sleigh,
1974)
By the Paramecium moves forward and travels
in a straight line. However, it is possible for it to
move backward if there are obstacles such as a
solid object or a predator. This phenomenon is
called avoidance reaction, which generally
causes the cilia to start beating in the opposite
direction. This results in a stop and a spiral
movement in order to return in swimming
forward. (Vidyasagar, 2016).
The ciliate has no cell wall but instead has a
pellicle on which the cilia are attached.
Underneath the pellicle is the plasma membrane,
and under which is the myonema. Between the
bases of the cilia are trichocysts and toxicysts
which are used for defense and attack. To avoid
being engulfed by predators, these structures are
discharged as filaments composed of a cross-
striated shaft and a tip. (Hickman, 2013)
Role of the Contractile Vacuole
The contractile vacuole is the structure
responsible for osmoregulation of the cell. It
prevents the Paramecium from absorbing too
much water that could lyse the cell. Structurally,
it is fed by channels radiating from it and each
of them pulsates alternately. Also, the ciliate has
two of it, the anterior and posterior contractile
vacuoles. (Unger, 1925)
The discharge rate or pulsing of the contractile
vacuoles decreases in increasing solute
concentration in the environment. Paramecium
generally tolerates NaCl concentrations up to
40%. (Watcherman, 1953).
Digestion
In digestion, the feeding apparatus in
Paramecium consists of a shallow ciliated oral
groove, a ciliated tube which leads into the
body, and a bundle of fibers (esophageal fibers)
which extend from the tube nearly to the
posterior end of the body. The tube is composed
of an outer part (the vestibulum) and an inner
part (the pharynx). Food particles are taken
inside the oral groove which the beating of its
cilia causes food-laden water to be swept into
the oral groove from a distance in advance of the
anterior end. Then, the food particles pass
through the vestibule where some are rejected
and thrown out, but others pass into the
cytostome. The strong cilia lining the vestibule
causes this selection mechanism. (Mast, 1947)
When the food vacuole reaches a certain size the
post-buccal fibers clasp the food vacuole and it
is pinched off by them and started on its course.
Cytoplasmic streaming or cyclosis carry the
food vacuoles along a definite course which is
functionally equivalent to a digestive tract.
[REF]
During cyclosis, digestion occurs by enzymes
secreted by protoplasm into the vacuoles. In
digestion, proteins are changed into amino acids,
carbohydrates into soluble sugars and glycogen,
and fats are probably also digested. The contents
of food vacuoles are at first acidic (pH about 4)
and then become alkaline, major digestion
occurs during the alkaline phase. The undigested
matter is egested through the cytopyge with
some force. (Sikora, 1981)
Experimentally, cyclosis can be manifested
through using Congo red. Food vacuoles will
first turn red due to acidic reaction of enzymes,
then change to blue due to alkaline reaction. The
color change signals the influx of the digestive
enzymes into the food vacuole in which a
decrease in acidity can be observed. (Nisbet,
1984)
The Reaction of Paramecium to Various
Stimuli
Thigmotaxis
Thigmotaxis is the most frequent stimuli
encountered by Paramecium. In an experiment
demonstrated by Jennings in 1931, Paramecium
showed high sensitivity on its anterior end when
a glass tip comes into contact with the body
(Witcherman, 1953). When the anterior end of a
Paramecium bumps into an obstacle - in the
experiment, cotton fibers - it moves backwards
and spins to another direction in order to return
to its forward movement. This mechanism is
called avoidance reaction which causes the
spiral movement of the Paramecium. The
response is also consistent with the observations
of Naitoh and Eckert in 1968: the stimulus of a
glass tip on the anterior portion resulted to
reversal in ciliary action and increased
permeability of cell membrane to calcium ions.
However, when the posterior part of the ciliate
comes into contact with a solid barrier, it
continues to move forward. Naitoh and Eckert
also observed the same response: an increase in
ciliary action in a forward direction and
increased permeability of cell membrane to
potassium ions. This avoidance reaction is
important for the ciliate in order to survive
against harmful obstacles such as predators and
dangerous chemicals. (Vidyasagar, 2016).
Chemotaxis
Paramecium cells exhibit different responses
towards different pH levels. In the experiment, it
demonstrated positive chemotaxis towards
sodium bicarbonate, 3% NaCl, and 5% sucrose
while negative chemotaxis towards 1M HCl, and
acetic acid. Theoretically, the ciliate should
move towards sucrose, and acetic acid. It acts in
this way because sucrose is a food source.
According to Chase & Glaser (1930), the more
H+ molecules are present, the acceleration
would be more faster. This implies that
hydrochloric acid and acetic acid would cause
an increased acceleration of ciliary beating in
the ciliate. On the other hand, Paramecium
should move away from sodium bicarbonate and
sodium chloride which induces the cell to
become hypotonic, causing cell lysis.
Geotaxis
The response of Paramecium to gravity is
affected by many factors thus, the geotaxis of
Paramecium is not a simple mechanism. In
typical conditions, Paramecium was observed to
be negatively geotactic - the natural response of
an organisms to swim upward in a container, but
the experimental result recorded the inverse.
Observations of Witcherman showed that
Paramecium tend to aggregate in a ring just
below the meniscus of the water. Even after
inverting the tubes or centrifuging cultures of
Paramecium, the single-celled protist continued
to swim upwards with a considerable speed. The
negative geotaxis of Paramecium is dependent
on a few factors: partial pressure of dissolved
oxygen, sexual reactivity, and nutrition
(Witcherman, 1953).
Phototaxis
Paramecium exhibits negative geotactic
response, but the intensity of light and the
natural upward swimming of Paramecium are
not related. Only one species of Paramecium (P.
bursaria) is positively phototactic, and other
species do not respond to light under natural
conditions. The experimental result observed a
positive phototaxis. When light intensity
increased sharply, Paramecium are generally
negatively phototactic. When exposed to other
forms of electromagnetic radiation like radium
and x-rays, the movement of Paramecium is
greatly accelerated. After prolonged exposure,
the movement of Paramecium decreases below
normal and an action follows after that will
result to death. Paramecium immediately reacts
negatively when exposed to ultraviolet light
(Witcherman, 1953).
Galvanotaxis
Paramecium was shown to have responses to
electric stimuli, but the causes of the responses
are still obscure. When a culture of Paramecium
is subjected to an electric current from a
galvanic cell, all cells swim towards the cathode
or the negative electrode. When the electrodes
are interchanged, Paramecium will swim toward
the new position of the cathode. The speed of
the response depend on the strength of the
electric current. According to Verworn who
observed the behavior of Paramecium from
1889 to 1915, Paramecium moves sluggishly or
does not respond well under weak current. When
the current is increased, the movement of
Paramecium is faster and sharper to the cathode.
Currents much stronger than that result to
Paramecium immobilized with the anterior end
pointed to the cathode. Further increase in
current will make the cells shorter and thicker
and began swimming towards the anode until
one end of the cell burst open and Paramecium
disintegrates (Witcherman, 1953).
*LUDLOFF PHENOMENON
CONCLUSION
The Paramecium is an unicellular ciliate that
usually lives in murky or brackish waters. The
morphology of the ciliate was observed under a
light microscope and was described as an
organism with a sharp, anterior end and a blunt,
posterior rear. Its body is covered by an elastic
material called the pellicle, which acts as a cell
membrane. Two types of nuclei can be seen
inside the cytoplasm, the macronucleus and the
micronucleus; the former is used for controlling
the actions of the cell while the latter is used for
sexual recombination. An eyespot is present for
the detection of light, and large contractile
vacuoles that function of osmoregulation where
they filter excess water out of the cytosol. As
shown in Table 1, the higher the salt
concentration of the solution the less the need
for the contractile vacuole to expel the water
since the water would freely go out of the cell.
An oral groove can be sometimes observed
which functions for feeding of the organism. It
has a flagella which is used for locomotion in
which it rotates counter-clockwise, making the
Paramecium move in the same direction. Cilia
are also present on its body where they also aid
in movement by beating in unison in one
direction. This movement is defined as the
spiral movement of the Paramecium where it
spirals in a counterclockwise direction while
travelling on a straight line.
The ciliate was subjected to different external
stimuli in which its reactions to each was
observed. When subjected to physical obstacles,
the Paramecium would slightly retreat then
change direction when its anterior end was
bumped; it would move to the opposite direction
of the barrier when its posterior end was
bumped.
For chemotaxis it was observed that the
Paramecium was attracted to sodium
bicarbonate, sucrose and 3% NaCl while it
avoided acetic acid and HCl. Theoretically, it
should be attracted either to its food source or to
substances that have high proton concentrations
that would increase its acceleration; it would be
repulsed by chemicals that can cause hypotonic
solution which can lead to cell lysis, such as
NaCl and sodium bicarbonate.
The experimental effect of gravity on the
samples are contrast to its theoretical effect.
Ideally, the Paramecium should float to the top
of the container, but in the experiment they
aggregated at the bottom of the test tube. This
may be caused by other factors such as pressure,
vitality, and sexual reactivity.
In phototaxis it was seen that the Paramecium
reacts positively to light where it stayed in the
portion where light can seep in. However, in
increased light intensity it is theoretically said
that the ciliate would avoid exposure to that
location. It would also react negatively to UV
rays in which prolonged exposure would result
to death.
When subjected to an electric current the
Paramecium would swim to the cathode (-) side.
This observation is correspondent to the
theoretical observation that the ciliate would
swim to the cathode, and the higher the current
the faster it would swim. However, high
voltages would lead to the cell lysis of the
Paramecium.
REFERENCES
Blake, J. R., & Sleigh, M. A. (1974). Mechanics
of Ciliary Motion. Biological Reviews,
49(1), 85-125. doi:10.1111/j.1469-185
X.1974.tb01299.x
Chase, A. M., & Glaser, O. (1930). FORWARD
MOVEMENT OF PARAMECIUM AS A
FUNCTION OF THE HYDROGEN ION
CONCENTRATION. The Journal of
General Physiology, 13(6), 627–636.
Hickman, C. (2013). Integrated Principles of
Zoology, 16th ed. New York: McGraw Hill
Education.
Randall, D. J., Burggren, W. W., French, K., &
Eckert, R. (2002). Eckert animal
physiology: Mechanisms and adaptations.
New York: W.H. Freeman and Co.
Reece, J.B., Urry, L.A., Cain, M.L., Wasserman,
S.A., Minorsky, P.V., & Jackson, R.B.
(2014). Campbell Biology (9th ed.). San
Francisco, CA: Benjamin Cummings.
Sikora, J. (1981). Cytoplasmic streaming in
Paramecium. Protoplasma. 109. 57-77.
10.1007/BF01287630.
Vidyasagar, A. (2016). What Is a Paramecium?
Retrieved September 02, 2017, from
https://www.livescience.com/55178-
paramecium.html
Mast, S. O. (1947). The Food Vacuole in
Paramecium. The Biological Bulletin, 92(1),
31-72. doi:10.2307/1537967
Nisbet, B. (1984). Feeding in the Ciliophora.
Nutrition and Feeding Strategies in
Protozoa, 128-155. doi:10.1007/978-94-
011-6555-6_8
Unger, W. B. (1925). The relation of contractile
and food vacuoles to rhythms in
Paramecium. Experimental Biology and
Medicine, 22(6), 333-334.
doi:10.3181/00379727-22-162
Witcherman, R. (1953). The Biology of
Paramecium. Retrieved from
https://babel.hathitrust.org/cgi/pt?
id=mdp.39015018196421;view=1up;seq=9