Evaluation of Thermophile Algae in Horizontal Photo-Bioreactor

83 Pages·2013·1.65 MB·English

by  Olivier Schmied

Evaluation of Thermophile Algae in Horizontal Photo-Bioreactor

by

Olivier Schmied

BioResource and Agricultural Engineering

BioResource and Agricultural Engineering Department

California Polytechnic State University

San Luis Obispo

2013
SIGNATURE PAGE

TITLE : Evaluation of Thermophile Algae in Horizontal Photo-
Bioreactor

AUHTOR : Olivier Schmied

DATE SUBMITTED : May 17, 2013

_Dr. Andrew Holtz______________ ______________________________
Senior Project Advisor Signature

______________________________
Date

_Dr. Ken Solomon______________ ______________________________
Department Head Signature

______________________________
Date

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ACKNOWLEDGEMENTS

Acknowledgements listed in no particular order.

First, I would like to express my appreciation to Professor Melinda Keller who made this
project possible.

Second, I would like to express my appreciation to Dr. Brian Hampson who made the
FSN (Food Science & Nutrition) Lab available to conduct algae experiments to make this
project possible.

Third, I would like to express my appreciation to Dr. Andrew Holtz who offered
guidance in times of need.

Fourth, I would like to express my appreciation to Virgil Threlkel for assistance in Lab
during the construction phase of this project.

Fifth, I would like to express my appreciation to John King and employees at Combined
Power who funded Melinda Keller, as well as donated an algae sample to conduct this
project.

Sixth, I would like to express my appreciation to the CPCONNECT Pilling Endowment
whose funding allowed for this project to be completed.

Sixth, I would like to express my appreciation to my friends and family for supporting me
throughout my academic career at California Polytechnic State University.

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ABSTRACT

The project investigator designed and built two algae photo bioreactors, one of which was
integrated into a test pond where Melinda Keller conducted heat transfer experiments
with fluctuating temperatures. The other was used as a control experiment. The project
investigator decided to test a thermophilic strain of algae in this system. To obtain
optimum culture growth, the project investigator implemented an air mixing system into
the photo bioreactor. The air mixing system promotes gas exchange between the fluid and
the algae cells.

After algae were cultured in the Food Science and Nutrition Lab at Cal Poly, the project
investigator introduced thermophilic algae into both photo bioreactors at Melinda
Keller’s test pond and control tank. Data from all the cultures were used to create growth
curves. The control experiment grew slowly and appeared to be stressed constantly due to
lower temperatures. The photo bioreactor in Melinda Keller’s test pond responded well to
the temperature fluctuations and experienced healthy growth. The cell counts showed
steady growth to a maximum culture density. The project investigator concluded that a
thermophilic stain of algae is suitable for the temperature environment being created by
Melinda Keller’s heat transfer experiments.
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DISCLAIMER STATEMENT

The university makes it clear that the information forwarded herewith is a project
resulting from a class assignment and has been graded and accepted only as a fulfillment
of a course requirement. Acceptance by the university does not imply technical accuracy
or reliability. Any use of the information in this report is made by the user(s) at his/her
own risk, which may include catastrophic failure of the device or infringement of patent
or copyright laws.

Therefore, the recipient and/or user of the information contained in this report agrees to
indemnify, defend and save harmless the State its officers, agents and employees from
any and all claims and losses accruing or resulting to any person, firm, or corporation
who may be injured or damaged as a result of the use of this report.
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TABLE OF CONTENTS

SIGNATURE PAGE ..................................................................................................................... ii
ACKNOWLEDGEMENTS.......................................................................................................... iii
ABSTRACT .................................................................................................................................. iv
DISCLAIMER STATEMENT ...................................................................................................... v
LIST OF FIGURES .................................................................................................................... viii
INTRODUCTION ........................................................................................................................ 1
Background .......................................................................................................................................... 1
Justification ........................................................................................................................................... 1
Objectives .............................................................................................................................................. 2
LITERATURE REVIEW ............................................................................................................. 3
Background of Energy Crisis .......................................................................................................... 3
Introduction to Microalgae ............................................................................................................. 4
Microalgae Production Systems .................................................................................................... 5
Potential Uses for Algal Biomass .................................................................................................. 7
PROCEDURES AND METHODS .............................................................................................. 9
Algae Collection in Mammoth Lakes California ....................................................................... 9
Thermophilic Algae from Hot Springs. ................................................................................................. 9
CP Connect Grant. .......................................................................................................................................... 9
Mammoth Lakes Hot Spring. .................................................................................................................... 9
Design/Construction of Main Photo-Bioreactor Pond (PBR) System ........................... 10
Design/Construction of Control PBR ........................................................................................ 12
Procedures for Cultivating Algae ............................................................................................... 12
Stock algae culture. ..................................................................................................................................... 12
Nutrient Media ............................................................................................................................................. 12
Culture Scaling .............................................................................................................................................. 13
Procedures for Quantifying Culture Density ......................................................................... 13
1L Lab Culture Cell Count. ....................................................................................................................... 13
20L Lab Culture Cell Count ...................................................................................................................... 13
Control PBR Cell Count ............................................................................................................................. 13
80L PBR Cell Count. .................................................................................................................................... 13
RESULTS .................................................................................................................................... 15
Algae Growth Results ..................................................................................................................... 15
Mammoth Lakes Algae Collection .............................................................................................. 25
DISCUSSION .............................................................................................................................. 27
Maximum Synechococcus Culture Temperature ................................................................. 27
Maximum Culture Density ............................................................................................................ 27
Nutrient Depletion .......................................................................................................................... 28
Shepard’s Hot Spring, Mammoth Lakes CA ............................................................................ 28
Hot spring temperature. ........................................................................................................................... 29
Evidence of Algae Culture Stress ................................................................................................ 29
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Air Mixing System ............................................................................................................................ 31
Conclusion .......................................................................................................................................... 31
RECOMMENDATIONS ............................................................................................................ 32
Project Improvements ................................................................................................................... 32
Project Continuation ...................................................................................................................... 32
REFERENCES............................................................................................................................. 34
APPENDICES ............................................................................................................................. 36
APPENDIX A: CP CONNECT PROJECT PROPOSAL / AWARD ..................................... 37
APPENDIX B: SOLIDWORKS RENDERINGS / HARDWARE IMAGES ........................ 43
Control PBR Assembly ................................................................................................................... 44
Main PBR Assembly ........................................................................................................................ 46
Combined Power end cap. ....................................................................................................................... 46
Inner tube assembly ................................................................................................................................... 46
Outer tube assembly .................................................................................................................................. 47
Main PBR air mixing system ................................................................................................................... 47
APPENDIX C: BRISTOL MEDIUM RECIPE ....................................................................... 49
APPENDIX D: COUNTING CHAMBER INSTRUCTIONS ................................................ 51
APPENDIX E: ALGAE CULTURE DATA ............................................................................. 61
APPENDIX F: ALGAE CULTURE GROWTH STAGES ...................................................... 71

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LIST OF FIGURES

Figure 1: Gasoline cost increase since the closeout of the NREL in 1996. (EIA, 2013) .... 3
Figure 2: Diesel cost increase since the closeout of the NREL in 1996. (EIA, 2013) ........ 4
Figure 3. Illustration of carbon dioxide conversion by photosynthesis. (Bayless, 2000) ... 4
Figure 4: Advantages of using photo-bioreactors vs. open ponds. (Olsen, 2009) .............. 6
Figure 5: Temperature of Synechococcus compared to maximum rate of photosynthesis
(Castenholz, 1969). ..................................................................................................... 7
Figure 6. Shepard’s Hot Spring map, Mammoth Lakes CA. (Google Maps, 2013) ........ 10
Figure 7. Artificial test pond. ............................................................................................ 11
Figure 8. Main PBR with respect to water level. .............................................................. 11
Figure 9. Algae samples, Main PBR & Control PBR. ...................................................... 14
Figure 10. 5 stages of growth for micro-algae. (Lavens). ................................................ 15
Figure 12. 1L culture density, 1/25/13-2/12/13. ............................................................... 16
Figure 13. Culture pigment comparison. .......................................................................... 17
Figure 14. 19L culture density, 2/12/13-2/19/13. ............................................................. 17
Figure 15. 1L culture density, 2/21/13 - 3/16/13. ............................................................. 18
Figure 16. 19L culture density, 3/16/13 – 3/21/13............................................................ 19
Figure 17. 1L culture density, 3/16/13 – 4/5/13................................................................ 20
Figure 18. 20L culture density, 4/5/13 - 4/24/13. ............................................................. 21
Figure 19. Main PBR temperature cycle. .......................................................................... 22
Figure 20. 80L culture density, 4/22/13 - 5/14/13 ............................................................ 23
Figure 21. 20L culture density, 4/24/13 - 5/14/13. ........................................................... 24
Figure 22. 5.55L culture density, 4/30/13 - 5/14/13 ......................................................... 25
Figure 23. Project Investigator at Shepard's Hot Spring. .................................................. 26
Figure 24. Shepard's Hot Spring, Mammoth Lakes CA. .................................................. 26
Figure 25. Green algae collected at Shepard's Hot Spring. .............................................. 26
Figure 26. Death phase in 20L culture. ............................................................................. 27
Figure 27. Shepard's Hot Spring algae sample. ................................................................ 28
Figure 28. Control PBR culture stress. ............................................................................. 29
Figure 29. Stock culture. ................................................................................................... 30
Figure 30. Culture health of Main PBR. ........................................................................... 30
Figure 31. Main PBR air mixing system. ......................................................................... 31
Figure 32. Control PBR Components. .............................................................................. 44
Figure 33. Control PBR, air-mixing system. .................................................................... 44
Figure 34. Control PBR in control tank at M.E. project labs............................................ 45
Figure 35. Control PBR air-mixing example. ................................................................... 45
Figure 36. Inner and outer end cap. .................................................................................. 46
Figure 37. Inner tube assembly. ........................................................................................ 47
Figure 38. Outer tube assembly. ....................................................................................... 47
Figure 39. Air mixing system. .......................................................................................... 48
Figure 40. Main PBR in test pond. ................................................................................... 48
Figure 41. Stationary and death stage. .............................................................................. 72
Figure 42. Exponential and stationary stage. .................................................................... 73
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LIST OF TABLES

Table 1. Cell-count data, 1/25/13 - 2/12/13, 1 Liter ......................................................... 62
Table 2. Cell-count data, 2/12/13 – 2/19/13, 19 Liter ....................................................... 63
Table 3. Cell-count data, 2/21/13 – 3/16/13, 1 Liter ......................................................... 64
Table 4. Cell-count data, 3/16/13 – 3/21/13, 19 Liter ....................................................... 65
Table 5. Cell-count data, 3/16/13 – 4/15/13, 1 Liter ......................................................... 66
Table 6. Cell-count data, 4/5/13 – 4/24/13, 20 Liter ......................................................... 67
Table 7. Cell-count data, 4/22/13 – 5/14/13, 80 Liter ....................................................... 68
Table 8. Cell-count data, 4/24/13 – 5/14/13, 20 Liter ....................................................... 69
Table 9. Cell-count data, 4/30/13 – 5/14/13, 20 Liter ....................................................... 70

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