1. Ecology of Open Algae Ponds Rob McBride October 2014 Rob McBride October 2014

2. Sapphire Produces Valuable Products From Algae, Sunlight, & CO 2 SAPPHIRE ENERGY 1

3. Sapphire’s Primary Production Platform – Open Ponds SAPPHIRE ENERGY 2 Focused on the production of low cost biomass Open ponds are continuously operated year round Key challenge to continuous open operation is stability Pests primarily impact stability

4. Open Pond Stability SAPPHIRE ENERGY 3 Mechanical strategies Netting, filters Cultural strategies pH, Ammonia, Magnesium etc. Chemical strategies Chlorine, glyphosate, headline, etc. Biotic strategies Trophic interactions

5. An Example of a Sapphire Pest Management Strategy SAPPHIRE ENERGY 4

6. An Example of a Sapphire Pest Management Strategy SAPPHIRE ENERGY 5 1. Letcher, P. M., S. Lopez, et al. (2013). "Characterization of Amoeboaphelidium protococcarum, an Algal Parasite New to the Cryptomycota Isolated from an Outdoor Algal Pond Used for the Production of Biofuel." PloS one 8(2): e56232. 2. McBride, R. C., S. Lopez, et al. (2014). "Contamination Management in Low Cost Open Algae Ponds for Biofuels Production." Industrial Biotechnology 10(3): 221-227.

7. Why Are We interested in Open Pond Ecology? SAPPHIRE ENERGY 6 1. Shurin, J., R. Abbott, et al. (2013). "Industrial‐strength ecology: trade‐offs and opportunities in algal biofuel production." Ecology letters 16(11): 1393-1404.

8. In Open Algae Ponds, What is the Richness/Stability Relationship? SAPPHIRE ENERGY 7

9. Overview of the Data Collection SAPPHIRE ENERGY 8

10. Summary of the Generated Data SAPPHIRE ENERGY 9 Bacteria Eukaryotes

11. Summary of the Generated Data SAPPHIRE ENERGY 10 Bacteria Eukaryotes

12. Is Open Algal Pond Species Richness Related to Productivity/Stability? SAPPHIRE ENERGY 11

13. How Do Pond Management Practices Impact Species Richness? SAPPHIRE ENERGY 12

14. How Do Pond Management Practices Impact Species Richness? SAPPHIRE ENERGY 13 Bacteria Eukaryotes

15. Open Algae Pond Ecology – What are the Punchlines? SAPPHIRE ENERGY 14 There was a positive relationship between increasing species diversity and the productivity of the open algae pond. Increasing species diversity did not increase the variance of the open algae pond with this analysis. Process and management strategies can impact ecology, and these impacts can be lasting. While we are still far from engineering open pond algae ecosystems, these type of data suggest that open pond algae systems display some of the patterns observed in terrestrial ecosystems. We and others are working to construct stable consortia in the lab that have productivity and stability benefits, and hope to translate those to the field to realize the potential of the open pond platform for the continued production of low cost biomass.

16. Acknowledgements UCSD ­Steve Mayfield ­John Shurin ­Vineet Bafna ­Doruk Beyter Life Technologies ­Farzad Haerizadeh ­Pei-Zhong Tang ­Scott Becker ­Todd Peterson SDSU ­Yan Wei Lim Sapphire Energy ­Las Cruces – Crop protection Team (Kalli Lambeth), Harvest Team, Cultivation Team ­San Diego – Tony Hoang, Damla Bilgin, Craig Behnke SAPPHIRE ENERGY 15

17. SAPPHIRE ENERGY 16 Questions?