It's a basic knowledge about isolation of algae

1. MICROALGAL
TECHNOLOGY

2. SAMPLE COLELCTION AND ISOLATION
OF ALAGE
SUBASRI.M
I-MSC MICROBIOLOGY
DEPARTMENT OF MICROBIOLOGY
VIVEKANANDHA ARTS AND SCIENCE COLLEGE FOR WOMEN
VEERACHIPALAYAM, SANKARI, SALEM

3. CONTENTS
• Introduction
• Sample collection
• Isolation methods
• Conclusion

4. INTRODUCTION
• The algae belongs to the kingdom “Protista “and domain “eukaryote”
• There are at least 20,000 sps were identified.
• soil and water is the major source for the isolation of algae
• several Methods and basic culture media are developed in the late 1800 &
early 1900
• For the isolation of algae from soil & fresh water algae, temperature and
oxygen is selective factor of major importance
• The algae are found in water sources like pond water, fresh water, waste
water, marine water..... Etc.
• The algae are found in soil sources like terrestrial soil, aquatic soil, rock
soil, desert soil.

5. SAMPLE COLLECTION
• Most algae are collected using very simple and common tools.
• Depending on what type of algae is being collected determines the type of
tool needed.
• There are 2 main locations where algae live, suspended in the water or
associated with the bottom surfaces.
• Once samples are collected, they can be kept fresh in a refrigerator for a
couple of weeks or they can be preserved for long term storage.
• The preservative gluteraldehyde is most commonly used for this purpose.

6. Algae that suspended in the water
• The net is attached behind a boat, it is dragged in
the water, its acting like a filter, concentrating the
algae and allowing most of the water to pass
through the net, settle the algae in the bottom of
the net.
• A deep water sampler used to collect water
several meters below the surface.

7. Algae can be associated with sand
• Add a little To collect the algae associated with sand, the sand is
sucked up using a turkey baster or plastic pipette.
• Gravel is scooped up with a device such as a spoon.

8. General methods
1. Micromanipulation method
2. Centrifugal washing method
3. Spraying method
4. Streak plating method
5. Serial dilution method
ISOLATION OF ALGAE

9. 1.MICROMANIPULATION METHOD
• It is also called as micromanipulation or capillary
pipette aspiration
• Most common method for single cell Isolation
• The tip of a capillary or Pasteur pipette is narrowed by heating in the
fine flame
• Algal cells of interest is sucked up into the capillary pipette through its
tip, while viewing under an inverted microscope
• Sucked cells are transferred onto sterile medium
• Incubate
Procedure

10. Micromanipulation method

11. 2.CENTRIFUGE WASHING TECH
• Collection of microalgae using 10, 30 & 120 µm plankton nets
• Keep in 50 ml plastic bottle
• Mix with at least 20 ml sterile either sterile BBM/CM
• Bring quickly to the laboratory
• Mix with sterile water
• Put into 4 centrifuge tubes
• Centrifuge at 5000 rpm for 5 minutes
• Through the supernatant
Procedure

12. • Mix the residue with equal volume of sterile water using vortex
• Centrifuge again at 5000 rpm for 5 minutes
• Repeat this process for at least 3 times to make free from bacteria
• Streak the microalgae by loop on the agar plate

13. 3.SPRAYING METHOD
• Prepare the Petri dish and the agar should be poured into the plate and wait for
solidification
• Hold a petri plate about 18 inches from the touching tips of two Pasteur pipettes.
• One of these is attached to an airline via a hose, and mounted onto a ring stand.
• The other pipette is suspended tip-up into a container holding the algal mixture.
• The airflow from the first pipette creates a vacuum that draws a stream of algae-
containing liquid up from the container through the second pipette.
• The airflow also sprays the suspended algae through the air, where they can be
intercepted by the agar plate.
Procedure

14. Spraying method

15. 4.STREAK PLATE TECH
• Prepare Petri dish and agar medium
• Label the petri dish with all important information, such as your name,
date, which media, time, sample name must include.
• The agar should be poured into the Petri dish and wait for solidification
• After solidification to pick up the sample, you can use either a metal loop
or disposable plastic loops.
• A loopful of sample is streaked on the first quadrant in a back-and-forth
motion on the agar plate.
Procedure

16. • Sterilise the loop by heating it in the Bunsen burner if using a metal loop.
• Streak the other three quadrants by a similar method.
• Close the lid of the plate after streaking, and store the dish upside
down in an incubator with optimal temperature.

17. Microalgae isolation by the streak plate

18. 5.SERIAL DILUTION METHOD
• The sample/culture is taken in a flask.
• Take six test tubes, each with 9 ml of distilled water are taken.
• The tubes are named as 10-1,10-2.......10-6.
• Now, 1 ml of sample is taken from flask through sterile pipette, added to the
1st test tube(10- 1), mix it well
• 1 ml of mixture is taken from the 10-1 dilution and add to the next test tube
(10-2)
• The same process is then repeated for the remaining tube
Procedure

19. • Chu 10 medium or beneck’s broth media is prepared (different dilutions)
in seperate conical flask & Sterilize at 121’c for 15-20 min
• After Sterilization, add 1ml of diluted water sample to conical flask (flask
containing chu 10 media )
• Incubate for 15 -20 days
• After incubation observe the flask for the growth of alg

20. Serial dilution method

21. • Isolation of algae from soil and water is very important for
environmental and industrial purposal
• Production of biofuel, pigment, single cell protein.
• Local weather is one of the major limiting factors when determining
which microalgae strains can be grown quickly in an established area
• Through this work certain species like Scenedesmus had found to have
scope for the successful production of biomass for biodiesel feedstock
in massive scale..
CONCLUSION