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Microalgae cultivation in different 𝒑 𝑯
,
temperature and media for
lipid production
Dakshayini Jayarama Reddy , Ravi kuma...
1. Introduction
- What is algae, biofuel and fossil-fuel in very simple
concept ?
- Why we should focus on biofuel ?
- How...
Introduction : What ? Why ? How ?
 Algae – simple plant like photosynthesizing organisms
# Microalgae (i.e., unicellular)...
Algae GOOD for Biodiesel Production
low cost,
high growth and
high biomass production rates.
Introduction : What ? Why ? H...
Introduction
WASTE
CO2
SALINE
WATER
NON-FOOD
RESOURCE
Waste
water
treatment
PROTEIN
&
CARBS
=40%
NON PRODUCTIVE LAND
: Wha...
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
50 100 200
600
1200
5000
GALLON/ACRE/YEAR
ENERGY CROPS
Comparison of oi...
: What ? Why ? How ?
National
Security ???
Introduction
: What ? Why ? How ?Introduction
Fossil fuels
&
Global Warming
!!!
∑GHG
TEMPERATURE
INCREASE
NEGATIVE
EFFECTS
Introduction
Price, mines and Demand ???
: What ? Why ? How ?
Trans-esterification (biodiesel formation)
Biodiesel
Introduction
Algal Oil Alcohol
Alcohol
Alcohol
Esters Glycerol
: What...
: What ? Why ? How ?Introduction
10 gm/L 100 gm/L1 gm/L
Mass production, Harvesting & Processing!!
Methods
Collection
Identification
Growth studies
Biomass Determination
Optical density
Thin Layer Chromatography
FLOW-CHAR...
1. Collection
Samples from the pond of BBMP park, Banglore
1. RR Nagar
2. Jaya Nagar
3. Bhuvaneshwori Nagar
Materials/Meth...
2. Screening of algae isolates
microalgae samples were spread on the plates
containing standard Blue Green 11 (BG-11) medi...
3. Identification
algae identification field Guide
by
Huynh and Serediak, 2006
Materials/Methods I Results & Discussion I ...
4. Preparation of Growth media and inoculation
Standard Blue Green-11 media according to Kuhl,
1964
Autoclaved at 121℃, 15...
5. Growth Studies- Qualitatively
 Measure Optical density at 540 nm against media
as a blank.
 The procedure was repeate...
6. Growth Studies- Quantitatively
 Harvested by centrifugation at 3000 rpm for 10 min.
 Noted wet biomass in g/L.
 Biom...
7. Effects of variables on lipid content - Analytically
Thin layer chromatography
based on a multistage distribution proce...
DA B C
1
2
?Origin
Solvent Front
x
y
Retention factor Value
Same or near 𝑹 𝒇 Value
means like compounds !!!
𝑅𝑓 =
𝑌
𝑥
1. Sp...
Physico-chemical parameters of algal water samples found
Highest that of RR nagar pond.
Huynh and Serediak, 2006
Chlorella...
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
25 30 35
0.43
0 0
0.36
0 0
OD
TEMPERATURE
Growth pattern vs. Temperatures
Chlor...
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
8 9 10
0.3
0.32
0.08
0.27
0.24
0.03
OD
PH
Growth pattern of algae in different 𝑷^𝒉
Chlor...
0
0.5
1
1.5
2
2.5
3
Beneck's Rao's Zarrouk's
2.94
0.56 0.53
0.29
0.73
0.22
OD
MEDIA
Growth pattern of algae in different m...
Materials/Methods I Results & Discussion I Conclusion
35.8
13.3
Wet Biomass in g/L
Chlorella Chladophora
4.45
1.2
Dry Biom...
•By Nile red staining
Chlorella sp. With lipid droplets inside the cell,
whereas Chladospora sp. without.
& thus TLC was p...
4. THIN LAYER CHROMATOGRAPHY
Materials/Methods I Results & Discussion I Conclusion
C-14-16-18
Zarrouk’s Rao’s
Beneck’s
pH ...
 The Chlorella sp. had a highest growth & biomass
at 25 ℃ (OD- 0.43)
𝒑 𝑯
-9 (OD-0.32)
Beneck's media (OD-2.94)
Materials/...
SUSTAINABILTY
GREEN JOBS AT HOME
OIL INDEPENDENCE
STRONGER ECONOMY
GREATER ENERGY SECURITY
HARMONICAL ENVIRONMENT
ACKNOWLEDGMENTS
Proff Dr. Sangeeta Rajbhandari
Assist. Dr. Giri Pd. Joshi
Tribhuwan University, Nepal
Thank you!!!
Microalgae cultivation in different pH, Temperature and Media
Microalgae cultivation in different pH, Temperature and Media
Microalgae cultivation in different pH, Temperature and Media
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Microalgae cultivation in different pH, Temperature and Media

Biotechnology advances in Algal Biofuel, one of the promising field for near future! This global warming due to our urgent necessary uses for all aspects of life, whether it is for living or just for recreation, we use fossils as our fuels. Even all your kitchen stuffs, from utensils to plastic containers... from LPG gas to your perfumes, medicines and cosmetics, all and all, directly or indirectly during their processings, uses this carbon emitting FOSSILS FUELS.
On the other opposite hand, we could use biofuel like Microalgae and various others.

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Microalgae cultivation in different pH, Temperature and Media

  1. 1. Microalgae cultivation in different 𝒑 𝑯 , temperature and media for lipid production Dakshayini Jayarama Reddy , Ravi kumar Krishnappa & Girisha Sirangala Thimmappa, Presentation based on Lipid production for Bio-fuel generation related research International Journal of Life Sciences 8 ( 2 ) : 2 0 1 4 ; 1 3 - 1 7 Bir Bahadur Thapa( M.Sc. I sem) Central Department of Botany Tribhuwan University Nepal
  2. 2. 1. Introduction - What is algae, biofuel and fossil-fuel in very simple concept ? - Why we should focus on biofuel ? - How is that possible ? 2. How researchers carried their research ? & 3. What they concluded from research ?
  3. 3. Introduction : What ? Why ? How ?  Algae – simple plant like photosynthesizing organisms # Microalgae (i.e., unicellular) – in micrometres. # Macro-algae (i.e., Seaweeds) – in inches.  Bio-fuels – fuels obtained from living plants!! - are carbon neutrals - renewable  Fossil-fuels – from dead’s of millions years ago!!!
  4. 4. Algae GOOD for Biodiesel Production low cost, high growth and high biomass production rates. Introduction : What ? Why ? How ?
  5. 5. Introduction WASTE CO2 SALINE WATER NON-FOOD RESOURCE Waste water treatment PROTEIN & CARBS =40% NON PRODUCTIVE LAND : What ? Why ? How ?
  6. 6. 0 500 1000 1500 2000 2500 3000 3500 4000 4500 5000 50 100 200 600 1200 5000 GALLON/ACRE/YEAR ENERGY CROPS Comparison of oil contents
  7. 7. : What ? Why ? How ? National Security ??? Introduction
  8. 8. : What ? Why ? How ?Introduction Fossil fuels & Global Warming !!!
  9. 9. ∑GHG TEMPERATURE INCREASE NEGATIVE EFFECTS
  10. 10. Introduction Price, mines and Demand ??? : What ? Why ? How ?
  11. 11. Trans-esterification (biodiesel formation) Biodiesel Introduction Algal Oil Alcohol Alcohol Alcohol Esters Glycerol : What ? Why ? How ? TAGs
  12. 12. : What ? Why ? How ?Introduction
  13. 13. 10 gm/L 100 gm/L1 gm/L Mass production, Harvesting & Processing!!
  14. 14. Methods Collection Identification Growth studies Biomass Determination Optical density Thin Layer Chromatography FLOW-CHART Screening Micro-algal Culture
  15. 15. 1. Collection Samples from the pond of BBMP park, Banglore 1. RR Nagar 2. Jaya Nagar 3. Bhuvaneshwori Nagar Materials/Methods I Results & Discussion I Conclusion
  16. 16. 2. Screening of algae isolates microalgae samples were spread on the plates containing standard Blue Green 11 (BG-11) medium Continuous subculture till the pure culture is obtained. Materials/Methods I Results & Discussion I Conclusion Incubated for 10-15 days 16:8 hours light/dark photoperiod.
  17. 17. 3. Identification algae identification field Guide by Huynh and Serediak, 2006 Materials/Methods I Results & Discussion I Conclusion
  18. 18. 4. Preparation of Growth media and inoculation Standard Blue Green-11 media according to Kuhl, 1964 Autoclaved at 121℃, 15 minutes & cooled Used as an inoculation And triplicate experiments Materials/Methods I Results & Discussion I Conclusion
  19. 19. 5. Growth Studies- Qualitatively  Measure Optical density at 540 nm against media as a blank.  The procedure was repeated for 10 days at regular time intervals of 24 hours. Materials/Methods I Results & Discussion I Conclusion
  20. 20. 6. Growth Studies- Quantitatively  Harvested by centrifugation at 3000 rpm for 10 min.  Noted wet biomass in g/L.  Biomass dried in a hot air oven at 50℃ overnight.  Noted dry biomass in g/L. Materials/Methods I Results & Discussion I Conclusion
  21. 21. 7. Effects of variables on lipid content - Analytically Thin layer chromatography based on a multistage distribution process. Materials/Methods I Results & Discussion I Conclusion
  22. 22. DA B C 1 2 ?Origin Solvent Front x y Retention factor Value Same or near 𝑹 𝒇 Value means like compounds !!! 𝑅𝑓 = 𝑌 𝑥 1. Spotting the TLC Plate standard lipids (𝑪 𝟏𝟒, 𝑪 𝟏𝟔 & 𝑪 𝟏𝟖 ) & Subject lipids 2. Development hexane & chloroform, 9:1 (v/v). 3. Visualization iodine chamber 4. The 𝑹 𝒇 value was calculated.
  23. 23. Physico-chemical parameters of algal water samples found Highest that of RR nagar pond. Huynh and Serediak, 2006 Chlorella sp. & Chladophora sp. Materials/Methods I Results & Discussion I Conclusion 1. Screening & Identification
  24. 24. 0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 0.4 0.45 25 30 35 0.43 0 0 0.36 0 0 OD TEMPERATURE Growth pattern vs. Temperatures Chlorella Chladophora Materials/Methods I Results & Discussion I Conclusion 2. OPTICAL DENSITY
  25. 25. 0 0.05 0.1 0.15 0.2 0.25 0.3 0.35 8 9 10 0.3 0.32 0.08 0.27 0.24 0.03 OD PH Growth pattern of algae in different 𝑷^𝒉 Chlorella Chladophora Materials/Methods I Results & Discussion I Conclusion 2. OPTICAL DENSITY
  26. 26. 0 0.5 1 1.5 2 2.5 3 Beneck's Rao's Zarrouk's 2.94 0.56 0.53 0.29 0.73 0.22 OD MEDIA Growth pattern of algae in different media Chlorella Chladophora Materials/Methods I Results & Discussion I Conclusion 2. OPTICAL DENSITY
  27. 27. Materials/Methods I Results & Discussion I Conclusion 35.8 13.3 Wet Biomass in g/L Chlorella Chladophora 4.45 1.2 Dry Biomass in g/L Chlorella Chladophora 3. BIOMASS ESTIMATION
  28. 28. •By Nile red staining Chlorella sp. With lipid droplets inside the cell, whereas Chladospora sp. without. & thus TLC was performed only for lipids from Chlorella. Materials/Methods I Results & Discussion I Conclusion 4. THIN LAYER CHROMATOGRAPHY
  29. 29. 4. THIN LAYER CHROMATOGRAPHY Materials/Methods I Results & Discussion I Conclusion C-14-16-18 Zarrouk’s Rao’s Beneck’s pH 8 pH 9 pH 10 pH 25℃
  30. 30.  The Chlorella sp. had a highest growth & biomass at 25 ℃ (OD- 0.43) 𝒑 𝑯 -9 (OD-0.32) Beneck's media (OD-2.94) Materials/Methods I Results & Discussion I Conclusion
  31. 31. SUSTAINABILTY GREEN JOBS AT HOME OIL INDEPENDENCE STRONGER ECONOMY GREATER ENERGY SECURITY HARMONICAL ENVIRONMENT
  32. 32. ACKNOWLEDGMENTS Proff Dr. Sangeeta Rajbhandari Assist. Dr. Giri Pd. Joshi Tribhuwan University, Nepal
  33. 33. Thank you!!!

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