Jeffrey Yau and Christina George Manhasset Science Research 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

Background 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

Mostly due to car emissions and industrial factories. (Hopwood, 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.

Raceway Pond Design http://www.agric.wa.gov.au/content/SUST/BIOFUEL/110407_Biodieselfrommicroalgae.pdf A raceway pond is made of a closed loop recirculation channel (Chisti, 2007)

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

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

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

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

Tubular Photo Bioreactor design for algal cultures 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

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

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

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

Hypothesis 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

Photo Bioreactor utilizing Airlift Bioreactor 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 utilizing Airlift 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 around 70 degrees Fahrenheit and cultured with a 10% Algal growth medium. Raceway pond utilizing and producing the algae’s energy 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)