Reviewer Comments - EERE

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2011 Algae Platform Review – Reviewer Comments Reviewer Comments are direct transcripts of commentary and material provided by the Platform’s Review Panel. They have not been edited or altered by the Biomass Program. driven by both the needs of the laboratory analysis and the capabilities of the producer. Clearly the best choices for nucleic acid preservation are flash freezing harvested biomass and shipment on dry ice. Alternatively the samples can be solubilized in Trizol and shipped on dry ice which will also preserve nucleic acids. Harvesting methods may impact nucleic acid content of the samples so these need to be evaluated. Frozen shipment methods do not generally preserve cellular integrity and do not allow for down stream physical separation methods. In many cases viruses are not captured by biomass harvesting methods. Shipment of "whole water" samples on ice has the benefit of retaining cellular integrity allowing downstream physical separation methods and can contain free virus. However as the reviewer correctly point out, changes in the samples can occur. As our nucleic acid analysis methods have improved, the need for physical separation has diminished and it is now apparent that a combination of frozen harvested biomass and filtrate (for viruses) is the optimal approach. 4. Critical Success Factors The project has identified critical factors, (including technical, business, market, regulatory, and legal factors) that impact the potential technical and commercial success of the project The project has presented adequate plans to recognize, address, and overcome these factors The project has the opportunity to advance the state of technology and impact the viability of commercial algal biomass feedstock supply and conversion, through one or more of the following: i. Cross-Cutting Analysis (ex. economic analysis, sustainability analysis, resource assessments, risk assessments) ii. Feedstock Supply R&D (ex. biology, cultivation, resource use, biomass characteristics, harvesting/dewatering) iii. Downstream Refining R&D (ex. extraction, conversion, fuel, products, fuel/product infrastructure and end-use) iv. Environmental sustainability (example: water use, GMOs, energy consumption) Reviewer Comments Reviewer: 1 Criteria Score: 5 These seem to be reasonably outlined. Reviewer: 2 Criteria Score: 6 The dipstick model is really important to practical work, so demonstration that this works against known pathogens/predators (e.g., viruses, bacteria, fungi, rotifers) is essential. Reviewer: 3 Criteria Score: 7 • Efficient and quantitative nucleic acid extraction and recovery. • Effective host and background subtraction Reviewer: 4 Criteria Score: 8 Still needed: Determine false positive/negative rates for various pests. Reviewer: 5 Criteria Score: 4 Critical success factors were identified but they seemed to focus primarily on the technical aspects of this approach rather than ultimate use of the technology. The project would be improved with additional input by those involved in large scale algal cultivation. Reviewer: 6 Criteria Score: 3 The project largely ignores other factors that can contribute to crashes that might have occurred. Page 154 of 223
2011 Algae Platform Review – Reviewer Comments Reviewer Comments are direct transcripts of commentary and material provided by the Platform’s Review Panel. They have not been edited or altered by the Biomass Program. Reviewer: 7 Criteria Score: 5 Beyond the success factors related to identifying pathogens quickly, there is the factor of defining actions that could conceivably be taken on commerical scale. Presenter Response We agree that the development of positive and negative controls and the determination of false positive and negative rates is a significant critical success factor. We are in the process of demonstrating the efficacy of our methods with known algal pathogens. We continue to develop appropriate collaborations and will focus on the development of collaborations with large scale producers. We have already interacted with and received letters supporting the importance and potential impact of our approach from Dr. Ami Ben-Amotz and from Sapphire Energy Inc, two of the leading and most respected algal biofuels developers in the world. Part of our strategy for developing these is an aggressive communications plan. We plan on presenting at national and international meetings early in the project to attract feedback from producers and to further advance the development of collaborations and other interactions. This project certainly focuses on the development of methods for the detection of biological agents to address the fact that current analytical methods are blind to broad classes of biological agents. However, this project does not, as the reviewer suggests, ignore the abiotic parameters that modulate the activity of biological agents or that are, in themselves the root causes of crashes. We will be collecting data on the physical and chemical parameters that are associated with the crash. We will use this data in addition to any putative biological agents, that may be present, to experimentally dissect and recapitulate the crash. 5. Technology Transfer and Collaborations Please comment on the degree to which the project adequately interfaces and coordinates with other institutions and projects to provide additional benefits to the Biomass Program, such as publications, awards, or others. Reviewer Comments Reviewer: 1 There should be a closer relationship with some pond operators. Reviewer: 2 This project is early on but has high potential for excellent technical transfer. Reviewer: 3 The goal of this project is to develop tools and methods that will be used to identify the etiological agents of pond crashes through the forensic analysis crash samples. Once identified, we will confirm by reproducing crashes in laboratory experiments. Reviewer: 4 Collaborations planned are only related to sample provision. Can anything be gained from collaborating with others on the genetics or rapid detection methods? Reviewer: 5 Technology transfer partners were not identified. Page 155 of 223
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Reviewer Comments - EERE

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