Isolation of algae
- 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
- 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
- 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