1) Researchers collected microalgae samples from ponds in Bangalore, India and screened them to obtain pure cultures of Chlorella sp. and Chladophora sp.
2) They studied the effect of temperature, pH, and growth media on microalgae growth and found that Chlorella sp. had the highest growth at 25°C, pH 9, and in Beneck's media.
3) Analysis showed Chlorella sp. had the highest biomass production and lipid content suitable for biodiesel production according to thin layer chromatography.
HMCS Max Bernays Pre-Deployment Brief (May 2024).pptx
Microalgae cultivation in different pH, Temperature and Media
1.
2.
3. 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
4. 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 ?
5. 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!!!
6. Algae GOOD for Biodiesel Production
low cost,
high growth and
high biomass production rates.
Introduction : What ? Why ? How ?
18. 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
19. 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.
21. 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
22. 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
23. 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
24. 7. Effects of variables on lipid content - Analytically
Thin layer chromatography
based on a multistage distribution process.
Materials/Methods I Results & Discussion I Conclusion
25. 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.
26. 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
29. 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
30. 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
31. •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
33. 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
34. SUSTAINABILTY
GREEN JOBS AT HOME
OIL INDEPENDENCE
STRONGER ECONOMY
GREATER ENERGY SECURITY
HARMONICAL ENVIRONMENT
Good Morning everyone. This is only an interesting day and I appreciate your patience and attention for a few moments!!
Hello and thank you so much for having me. I m delighted to be here. What a beautiful day it is and I m so happy to be sharing it with all of you.
The uni-cellar forms of algae, known as microalgae, have an extraordinary potential for cultivation as energy crops. In fact, algae can produce up to 300 times more oil per acre than conventional crops, such as rapeseed, palms or soybeans, and have a short harvesting cycle – one to ten days. They can be grown under conditions unsuitable for conventional crop production, reducing the burden on agricultural land .
Algae are the best suitable feedstock for biofuel production because of their low cost, high growth and high biomass production rates.
Much greater productivity than other energy crops---Non-food resource-----Can utilize saline water---Can utilize waste CO2 streams
Can be used in conjunction with waste water treatment---An algal bio-refinery could produce oils, protein, and carbohydrates!!!
Can be grown on marginal lands useless for ordinary crops •High yield per acre –have a harvesting cycle of 1–10 days
•Can be grown with minimal impact on fresh water resources
•Can be grown using flue gas from power plants as aCO2source
•Can convert a much higher fraction of biomass to oil than conventional crops, e.g. 60% versus 2-3% for soybean
National Security Not good to have energy dependence on foreign Countries.
Global Warming Fossil fuels release greenhouse gases ???
There is clear scientific evidence that emissions of greenhouse gases, such as carbon dioxide (CO2), methane (CH4) and nitrous oxide (N2O), arising from fossil fuel combustion and land-use change as a result of human activities, are disturbing the Earth’s climate.
The Price of Energy is Going UP -Oil reserves are depleting- World demand for energy is increasing.
Algae can be turned into a fuel by transesterification.--Ethanol is reacted with the algal oil, sodium ethanolate as the catalyst.--End product of this reaction are hence BIODIESEL, glycerol, sodium ethanolate!!!
Biofuels from land-rich tropical countries may help displace foreign petroleum imports for many industrialized nations, providing a possible solution to the twin challenges of energy
security and climate change.
To obtain a large amount of biomass component, major requirement is to select the best medium. The selection of the medium depends on the several factors which include the chemical composition of the medium to obtain the maximum growth of microalgae. Hence, the main goal of the present study is to evaluate the effect of different pH, temperature, and media on the growth and lipid content of isolated microalgae.
In general, measuring the optical density (OD) is a common method to quantify the concentration of substances (Beer-Lambert law), since the absorbance is proportional to the
concentration of the absorbing species in the sample.
The samples separately transferred to 50 mL falcon tubes--Centrifuged at 3000 rpm for 10 min--The supernatant collected, washed with 1% sodium chloride for 3 times.
The layer containing lipids with the solvent was evaporated in a rotary evaporator at 70 C under vacuum.
After evaporation the sample was dissolved in two solvents, chloroform & hexane--and subjected to thin layer chromatography.
like all chromatographic techniques, based on a multistage distribution process. This process involves: a suitable adsorbent (the stationary phase), solvents or solvent mixtures (the mobile phase or eluent), and the sample molecules.
2. Growth studies
Chlorella sp. & Chladospora sp. more growth at 25 C (OD- 0.43 and 0.36).
No growth was seen at 30C and 35C (r2= 0.997)
Chlorella sp. highest growth in Beneck's media (OD-2.94), minimum growth in Zarrouk's medium (OD-0.53) .
Chladospora sp. maximum growth in Rao's media (OD-0.73), minimum growth in Zarrouk's media (OD-0.22) and optimum growth in Beneck's media (OD -0.29) (r2= 1).
Hence, the present study reveals the effect of different physical and chemical conditions on the growth of Chlorella and Cladophora spp. Their growth can be improved further and there is a need to search for their low cost cultivation by using waste sources (agriculture and domestic wastes) to provide good sources of biomass to meet the primary requirements of alternative fuel production in future. The farming of algae, or algaculture, has the potential to address several of the world’s most demanding challenges, from rising oil prices and conflicts between the demand for biofuel and food, to the needs of a rapidly growing worldwide population. Biofuels, one of the most promising areas, can replace fossil fuels and help reduce the introduction of new carbon dioxide (CO2).
