Phagocytosis in Paramecium sp.

Mark Joel Aguit, Kiel Christian Alday, Ma. Kristelle Joyce Arellano, and Kristine Grace Austria
Department of Biological Sciences, University of Santo Tomas, España, Manila

Abstract
In the experiment conducted, the cell culture of Paramecium sp. was fed with Chlorella sp. and
graphite shavings to demonstrate the mechanism of phagocytosis and 1% India ink to
demonstrate the mechanism of pinocytosis. Microscopic examination reveals that the organism
utilizes its cilia to convey food into its gullet, forming a food vacuole that will be digested by
various enzymes in the cytoplasm until only wastes are left, which will be excreted to the cell’s
exterior through exocytosis. Results also show that food particle size and concentration play
factors towards food vacuole formation as smaller size and higher concentration lead to more
food vacuoles present in the cytoplasm with organisms fed with India ink having ten (10),
graphite shavings with eight (8), and Chlorella sp. with six (6). With this, the preferred food by
Paramecium sp. was identified to be India ink as it had the most number of food vacuoles
formed.
Keywords: phagocytosis, pinocytosis, Paramecium sp., food vacuole

Introduction laboratory work because it can easily be

Paramecium sp. is a large, ciliated,
unicellular protozoan that lives in freshwater
like streams, lakes and ponds (Katz &
Deterline, 2018). Like Tetrahymena sp., it is
an ideal specimen that can be used to study
the cell’s ciliated motion, morphology,
behavior and the structure and function of its
organelles (Wheatley et al. 1994). In
addition to that, this organism is also
considered as a suitable specimen for
grown and manipulated at large quantities.
Since Paramecia are unicellular organisms,
food particles cannot easily pass through the
cell through simple diffusion but instead are
ingested through the process of endocytosis.
Endocytosis is a fundamental process that
moves particles into the cell by engulfing it
with its plasma membrane and is usually
used to transport molecules that cannot pass
through the membranes passively. The
process includes the removal of receptors, Methodology
extracellular material, and membrane To demonstrate phagocytosis, a drop of
proteins and lipids from the surface of the Paramecium sp. suspension was first placed
cell (Picco & Kaksonen, 2018). on a slide then covered with a cover slip. A
drop of Chlorella sp. culture was placed on
Protozoans ingest their food through one side of the cover slip for the alga to be
phagocytosis or pinocytosis. Phagocytosis, able to move beneath it. Afterwards,
the condition of “cell eating”, is a type of phagocytosis of Chlorella sp. by
endocytosis which involves the process of Paramecium sp. has been microscopically
taking large or relatively large particles such observed under HPO. Another drop of
as bacteria, cell debris, or intact cells into Paramecium sp. suspension was again
the cell (Cooper, 2004). Meanwhile, placed on another glass slide and using a
pinocytosis is the condition of “cell cutter, graphite from a lead pencil was
drinking” and is a type of endocytosis that scraped until it fell on the drop of the
takes minute food particles or liquid Paramecium sp. culture. Cover slip was
substances, including water into the cell. placed over the glass slide and phagocytosis
This process results in a much smaller was again observed under HPO. Then, 1%
vesicle compared to phagocytosis and it India ink was mixed together with
does not require the vesicle to fuse with a Paramecium sp. suspension of equal volume
lysosome afterwards (Robibaro, 2011). in a microcentrifuge tube. The resulting
Phagocytosis in Paramecium sp. is a cell mixture was again observed under HPO,
behavior that can be directly observed using taking note of manner how the exposed cells
a compound microscope and can be to ink swim and eat compared to cells that
analyzed quantitatively by the number of have not been exposed to ink. After 10
vacuoles that form in a certain period of minutes of ink exposure, 50 𝜇L of the cell
time (Görtz & Fokin, 2009). In this and ink suspension was pipetted in a
experiment, the differences between microcentrifuge tube that contained 25 𝜇L of
phagocytosis and pinocytosis were 3% formalin solution. The new mixture was
distinguished, the pathway of food within mounted on a clean side and cover slip and
the said organism was traced, and the food the appearance and location of food
preference of the organism was identified.
vacuoles and cilia in the immobilized cells food, the vacuole becomes smaller in size
were observed. until all that is left are undigested particles,
which are released to the organism’s
Results and Discussion surroundings through its anal pore by
Upon performing the experiment, Figure 1 exocytosis. The process of phagocytosis of
shows the feeding mechanism through Paramecium sp. was demonstrated by
endocytosis of a unicellular protozoan, the feeding it with Chlorella sp. and graphite
Paramecium sp. As the organism is a filter shavings, which are large particles (>0.5𝜇m)
feeder that normally feeds on small whereas the process of pinocytosis was
organisms such as the algae, Chlorella sp., it demonstrated through the use of India ink,
utilizes its cilia to create water currents to which are very small particles.
collect and engulf food from its
surroundings towards its oral groove Figure 2 then illustrates the food vacuoles
(Ramoino, Diaspro, Fato, & Usai, 2012). formed as the Paramecium sp. cell culture
From the groove, collected food continues was fed with Chlorella sp. (A), graphite
through ciliary action down into the gullet shavings (B), and India ink (C). From this, it
wherein food vacuoles (or phagosomes) can be seen that the number of food
form when sufficient food has been stored. vacuoles formed from phagocytosis of
The formation of food vacuoles occurs as Chlorella sp. was the least at six (6), then
the organism’s plasma membrane creates a the phagocytosis of graphite shavings with
vesicle enclosing the food particles inside eight (8), and pinocytosis of India ink was
(Iwasa & Marshall, 2016). Vesicles are the greatest at ten (10). These results suggest
produced from the folds of the plasma that fewer food vacuoles are formed as the
membrane whose ends move until it fuses. food particle size such as Chlorella sp. (~2-
After the vesicle is formed, it pinches and 10 𝜇m) is bigger, while more food vacuoles
moves inside toward the organism’s are formed in the cytoplasm of the organism
cytoplasm where enzymes such as when the food particle size is smaller like
lysosomes enter it to digest the food, graphite shavings and India ink (0.05 𝜇m)
providing nutrients that will be absorbed by (Ramoino, 1997). The concentration of food
the organism (Ruppert, Fox, & Barnes, particles present is also a factor in the rate of
2004). As the enzymes continue to digest food vacuole formation as high
concentrations lead to more food vacuoles
and vice versa. Thus, both food particle size
and concentration are factors that determine
the food preference of Paramecium sp. From
the experiment, phagocytosis of India ink
had the most number of formed food
vacuoles, making it the preferred food of the
test organism.
Conclusion
In the experiment, phagocytosis was done by
Paramecium sp. as it engulfed the large food
particles (>0.5 𝜇 m) of Chlorella sp. and
graphite shavings while pinocytosis was
observed as Paramecium sp. takes in very
minute particles (<0.5𝜇 m) of the prepared
1% India ink. Also, the mechanism by which
these two processes occur starts from the
sweeping motion of the organism’s cilia to
collect food, engulfment of the food particle,
then formation of food vacuole, digestion by
enzymes, and finally excretion from the
cytoplasm through exocytosis. Lastly, India
ink was identified as the preferred food of
the organism as it had the most number of
food vacuoles formed at ten (10).
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