1. 1
TO STUDY THE GROWTH OF PARAMECIUM
SUBMITTED BY:
NAME :Gogisetti Saurabh
SESSION :2015-2016
ROLL NO. :14IMSILS042
SEMESTER NO. :Semester 4
PROJECT CARRIED OUT UNDER THE GUIDANCE OF
Dr. ASHUTOSH KUMAR
AT
INSTITUTE OF LIFE SCIENCES
AHMEDABAD UNIVERSITY
NAVRANGPURA, AHMEDABAD - 380009, GUJARAT
APRIL 2016

2. INTRODUCTION:
• It’s a unicellular eukaryotic protozoan which is ovoid, elongated and foot
shaped in structure.
• Whole cell is covered with cilia.
• Have closely spaced spindle shaped Trichocysts embedded deeply in cell
membrane that surrounds the organism.
• Deep oral groove is present, running from the anterior of the cell to its midpoint.
• Mainly live by heterotrophy, feeding on bacteria on bacteria & other small
organisms.
• Consists two nucleuses and contractile vacuoles.
2
Reproduction
Sexual
Conjugation occurs during unfavorable
conditions.
Asexual
Consists of transverse binary fission which
divides paramecium into two daughter cells.

3. MORPHOLOGICAL STRUCTURES
AND FUNCTIONS
3
Morphological Structure Function
Pellicle Membrane protecting the cell
Cilia Hair like structures to swipe in
food
Oral groove Collects food and directs it
Cell mouth Opening for food
Anal pore Disposes off waste
Contractile vacuole Contracts out excess water from
the cell
Radiating Canals Paths to contractile vacuoles
Trichocysts Used for defense
Gullet Forms food vacuoles

4. FOOD HABITS:
• They feed mainly on 1.Bacteria 2. Algae 3.Yeasts.
•When enough food is accumulated, it forms vacuole in the cytoplasm,
they begin to circulate through the cell.
• Enzymes present in the cytoplasm digest the food flowing in
vacuoles.
•When the vacuoles are fully digested, they are thrown out through
the anal pore with the waste contents into the environment.
• Food is detected using Trichocysts.
•Food vacuole travels from Anterior end to Posterior end. 4

5. REVIEW LITERATURE
The killer trait of Paramecium and its causative agents. By:Martina
Schrallhammer in December 2010.
•Paramecium provide the closest unicellular analog to the biological architecture
of multicellular species.
•Paramecium sometimes maintain endosymbiont relationship with some
microorganisms mainly with bacteria.
•In bacteria, species of Caedibacter are the species which make their host into a
killer trait.
•Caedibacter produce R-bodies which have- enrolling capacity, can form hollow
cylinder and is highly complex in structure.
•In presence of Caedibacter paramecia turn into killer species. The kill the
sensitive ones as per the natural selection.
•Infected stocks originate in: North America, Central America, Europe , Japan and
Australia.
5

6. REVIEW LITERATURE
The Ciliate Paramecium Shows Higher Motility in Non-Uniform
Chemical Landscapes. By : Giuffre et. Al in 2011.
•The used species here was Paramecium tetraurelia.
•They have trichocysts which detects light, temperature and variety of
attracting and repelling chemical substances.
•Chemosensitivty of Paramecium makes it a good biosensor for detecting
environmental pollutants in water.
•Motile organisms and cells sense their environment and react to it in a
directed motion by taxis.
•Known attractant is – Sodium Acetate [ C2H3NaO2] and repellent –
Potassium Ferrate [ K3[Fe(CN)6] .
6

7. METHODOLOGY
Aim: To study the growth of Paramecium.
Objective: Determining the growth of Paramecium using
different laboratory systems.
Materials needed:
Beakers
Protozoan pellets
Dryls buffer
Pipettes
20 µM filter
Distilled water
7
Wheat seeds
Multiplate reader (12
wells)
Stereomicroscope
Microscopic Slides
Incubator

8. METHODOLOGY
Media Preparation:
•In 100 mL of heated water 1 protozoan pellet was added.
•900 mL of distilled water was added to the 100 mL of protozoan
pellet mixture and autoclaved at 1210
C for 30 minutes.
•Dryls’ Buffer was prepared by mixing 1mM- Disodium
hydrogenphosphate, 1mM- Sodium Dihydrogen phosphate,
2mM- trisodium citrate, 1.5 mM-Calcium Chloride.
 Sodium citrate : Slow carbon source
Calcium chloride : responsible for locomotion /movement.
Dryls buffer: its allows Paramecium to grow/live for 6 days.
8

9. METHODOLOGY
Filtering /Isolating Paramecium from Stock:
1.The Paramecium from the stock was filtered using a 20µM filter into
three capped containers.
2.40 mL of media was taken and filtrated out into the media .
3.Grinded and boiled wheat seeds were added into the media.
4.The mixture was incubated for 1 day for good growth
5.The culture was filter out using 20µM filter. filter was washed in
Dryls buffer & observed under Stereomicroscope.
6.Paramecium was seen near the circumference of the container.
7.Using a 100 µL pipette , 50 µL of the Dryls buffer was taken on to a
slide and observed under a Stereomicroscope for their counting.
9

10. Case 1: Starvation:
1.1mL of pure Dryls buffer was added in 3 wells of 12 well plate, 4-5
Paramecium was added in it for overnight, without providing any media or
food or nutrients.
2.These paramecium was observed next day.
Case 2: Growing Paramecium in Media with Wheat:
1. Protozoan pellet media (200µL) was poured in each well of 12 well
plate.
2. 50µL from the Paramecium stock was added and observed under the
microscope on the slide.
3. 100µL Dryls buffer containing paramecium was added into the wells.
4. Grind and boil the wheat grain in an oven.
5.After this process soaked wheat grain was taken and poured into the wells,
left the culture for overnight.
10

11. Case 3:Studying growth using bacterial culture:
Preparing Media for Bacteria:Leuria Bertani:
•Mainly used in growing bacteria as it permits fast growth & good
growth yields for many species.
•Easy to prepare because it is easy to prepare & provide broad base
of nutrients.
•Components:
1.Tryptophan
2.Yeast extract
3.NaCl
•Taking 5g in 250 mL conical flask with 100ml distilled water,
keep it for autoclave for 30 minutes for 121˚C.
11

12. Growing Bacteria in the prepared media:
•In 100 mL of media 500µL of bacterial culture was added.
•Incubate this freshly made culture at 160 rpm at 37˚C , overnight
in incubator shaker.
•10 mL of the freshly prepared culturewas centrifuged at 3220 rcf
for 6 minutes.
•Pellet was mixed and centrifuged again10 ml of Dryls buffer for 2-
3 times.
•Vortex the pellet with Dryls buffer.
•10µL of the bacterial solution was added it to the well plates
containing Paramecium , media and wheat seeds.
•The bacterial strain used was E.coli K-14
12

13. Paramecium near the wheat grain
13
Paramecium in bacterial premises
RESULTS
• Paramecium in the wheat grain culture grew in number but less as compared to
the bacterial culture medium.
• In the wheat grain media , Paramecium were more near the wheat grain and
lesser as we move away from it.
• In bacterial culture media, they were swarming in the media as their food was
not intact.
• In the starvation media, very few paramecia were seen and they had slow
locomotion

14. CONCLUSION:
• Paramecium have an appetite for bacteria and if starved they become
immobile or slow moving. Sometimes their number also decreases in
starvation.
• As the Paramecium is bacterial grazer, it can used by water filtration
industries for filtering out bacterial strains from polluted waters.
• They can also be used to the keep the bacterial population in control,
balancing them in the ecosystem and in food chain.
14In bacterial culture ( 12-4-2016) On wheat seed (12-4-2016)

15. ACKNOWLEDGEMENTS:
• Dr. Rishi Shankar
Director, Institute of Life Sciences, Ahmedabad University
• Dr. Ashutosh Kumar
Assistant Professor, Institute of Life Sciences, Ahmedabad University
• Govind Saran Gupta
Senior Research Fellow, Institute of Life Sciences, Ahmedabad University
15Signature of the student Signature of the project supervisor