1. Developing New Working Protocols and Bioreactor Designs to Enhance Biomass Growth and Energy Yield in Schizochytrium limacinum and C.reinhardtii Jeffrey Yau and Christina George Manhasset Science Research

2. Background The use of bioreactors to combat the growing problem of greenhouse gases has been extensively studied in recent decades. The use of bioreactors to combat the growing problem of greenhouse gases has been extensively studied in recent decades. (Chisti, 2007) http://www.ieagreen.org.uk/newsletter/dec80/images/biofixation.JPG

3. The U.S. has reported a 3.3% increase of carbon dioxide emissions in the past year Mostly due to car emissions and industrial factories. (Hopwood, 2007) http://photos.mongabay.com/08/0423methaneglobal.jpg The red line shows the trend together with seasonal variations. The black line indicates the trend that emerges when the seasonal cycle has been removed.

4. Raceway Pond Design A raceway pond is made of a closed loop recirculation channel (Chisti, 2007) http://www.agric.wa.gov.au/content/SUST/BIOFUEL/110407_Biodieselfrom microalgae.pdf Original Design Modified Design Direction of Flow Taken: January 5, 2009

5. Tubular Photo Bioreactor Design Consists of straight transparent tubes either made out of glass or plastic (also known as solar collectors) Consists of straight transparent tubes either made out of glass or plastic (also known as solar collectors) (Chisti, 2007) http://www.agric.wa.gov.au/content/SUST/BIOFUEL/110407_Biod ieselfrommicroalgae.pdf Original Design Taken: January 5, 2009 Modified Design

6. Control: Airlift Design A self-contained bioreactor A self-contained bioreactor Utilizes a baffle to re-circulate the bacteria in suspension Utilizes a baffle to re-circulate the bacteria in suspension Taken on: December 21, 2008

7. Organism: S.limacinum Contains pigments for photosynthesis Contains pigments for photosynthesis Known to contain EPA, DHA, and omega-3 fatty acids Known to contain EPA, DHA, and omega-3 fatty acids Reliable source of oil production for biodiesels Reliable source of oil production for biodiesels (Kamlangdee, 2003) http://roweb.cityu.edu.hk/researchreport/2002-2003/Project/020.jpg

8. Organism: C.reinhardtii Contains an enzyme called hydrogenase that allows creation of hydrogen Contains an enzyme called hydrogenase that allows creation of hydrogen (Tiede, 2008) Ability to produce hydrogen under anoxic conditions Ability to produce hydrogen under anoxic conditions (Fouchard, 2005) http://en.wikipedia.org/wiki/Chlamydomonas_reinhardtii Magnified 3000X

9. Tubular Photo Bioreactor design for algal cultures Molina Grima; et al (2001): Experiment on tubular photobioreactors using P. tricornutum Molina Grima; et al (2001): Experiment on tubular photobioreactors using P. tricornutum Tested tubular diameter on the amount of sunlight that penetrated through the culture broth Tested tubular diameter on the amount of sunlight that penetrated through the culture broth

10. Polyunsaturated fatty acids production by Schizochytrium sp. Kamlangdee (2003): Experiment on polyunsaturated fatty acids production by Schizochytrium sp. Kamlangdee (2003): Experiment on polyunsaturated fatty acids production by Schizochytrium sp. Found single isolate reliable in production of DHA Found single isolate reliable in production of DHA

11. Hydrogen as Clean Fuel Via Continuous Fermentation by Anaerobic Photosynthetic Bacteria Continuous hydrogen production via fermentation of synthesis gas Continuous hydrogen production via fermentation of synthesis gas (Najafpour, 2003) http://www.vurup.sk/pc/vol45_2003/issue3-4/pdf/14.pdf

12. Purpose Therefore the purpose of this experiment was to create a bioreactor design that would enhance growth rate and energy yield in Schizochytrium limacinum and C.reinhardtii Therefore the purpose of this experiment was to create a bioreactor design that would enhance growth rate and energy yield in Schizochytrium limacinum and C.reinhardtii

13. Hypothesis Null Hypothesis: No difference will be found in either bioreactor design Null Hypothesis: No difference will be found in either bioreactor design Alternate Hypothesis: A significantly more amount of growth in both micro-organisms will occur in the Tubular Photobioreactor compared to the Raceway Pond Alternate Hypothesis: A significantly more amount of growth in both micro-organisms will occur in the Tubular Photobioreactor compared to the Raceway Pond

14. Methodology Problem: What is the most efficient design for a photo bioreactor to enhance the energy yield and growth rate of Schizochytrium limacinum and C.reinhardtii? Control Group: -Growth medium under normal conditions. -Growth rate in Airlift Bioreactor Independent Variable 1: Growth of Schizochytrium limacinum Dependent Variables: -Oil Extracted from Hexane -Hydrogen collected from C.reinhardtii -Three different sized tubes (0.01m, 0.012m, 0.019m) An ANOVA test will be used to statistically analyze the data (p<.05). The Scheffe post hoc test will be used. Schizochytrium limacinum will be obtained from atcc.org. The Glucose Yeast Extract Medium will contain 1g of Yeast Extract, 1g of Peptone, 5.0g of Glucose, and 1L of 15% Natural Seawater in a 1000mL Volumetric flask. The yeast extract, glucose, and the Natural Seawater will be obtained from Carolina Biological. Peptone will be obtained from Flinn Scientific. Growth Projection: 4 weeks Independent Variable 2: Growth of C.reinhardtii Photo Bioreactor Measurement of Biomass yield (once daily from start of experimentation) using Aquafluor Fluorometer C.reinhardtii will be obtained from Carolina, the Biological Vendor. The C.reinahrdtii will be cultured in a fresh water tank, with a 12 hour supply of light, and oxygen. The algae will be kept at 70 degrees Fahrenheit and cultured with a 10% Algal growth medium. Raceway pond Measurement of Biomass yield using Spectrophotometer (Wavelength at 610nm) Measurement of the effect of carbon dioxide on pH levels (Using Co2 Sensor on GLX and pH paper)