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. Critical Success Factors 2.71 1.28 7 1. Project Approach The project performers have implemented technically sound research, development, and deployment approaches and demonstrated necessary results to meet their targets The project performers have identified a project management plan that includes well-defined milestones and adequate methods for addressing potential risks. Reviewer Comments Reviewer: 1 Criteria Score: 3 The Approach is not really well-defined. The overall OBJECTIVES are outlined, and agreeable. There are 3 Phases in the Overview, but how the first will be addressed, and how decisions are to be made as to value in Phases 2 and 3, are not established. This important project would be helped with some thoughtful experimental planning. This should also extend to the dissemination of the results and especially their meaning. Reviewer: 2 Criteria Score: 2 This project aims to look at the potential for algal biofuels feedstock in ponds to become contaminated and toxic (e.g., with heavy metals, etc.). The presenter's concerns about confidentiality related to obtaining test samples from biofuels' research projects/facilities may be higher than justified. There is some concern about the inappropriate application of over-sophisticated technology in this project. Reviewer: 3 Criteria Score: 4 Same as the project by Yeager. Reviewer: 4 Criteria Score: 1 Slide 8 shows exposure of algae to 1-100 mg/L Cu followed by XANES analysis. What is the purpose of exposing algae to a well-known algae toxicant at extraordinarily high (1 mg/L) concentrations? I suspect these concentrations were used because XANES requires high concentrations for detection. What is the point of making measurements from single cells, rather than a bulk sample of biomass. This SSRL study does not seem to have any value. In addition, the cell toxicity work is not interesting until it is established that any important human exposure to algae toxins would occur. Of course the response is "How will we know the concentrations if we don't measure them?" However, start with the prevalence of toxinproducing species (expect trace to none based on current commercial algae production) and assess the potential routes of human exposure (workers touching culture water or blowdown water? -- risk is also minimal). Overall risk is negligible. The project proponents need to counter this type of opinion to justify the project. Study of media effects on algae, not humans, is a more pressing topic for researchers with these advanced skills and expertise. Reviewer: 5 Criteria Score: 2 The objective of the project is to perform a risk assessment for heavy metal and cell toxicity appropriate for large-scale algal ponds. The presentation completely failed to convince me that the approach was appropriate. Reviewer: 6 Criteria Score: 3 Very little to no justification was given as to the criteria for toxicity, nor the cells lines used. TAs the whole project hinges on these decisions why was the project started in the absence of major groups that study toxiity (CDC, Centers for Ocean and Human Health)? Page 168 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: 4 Because the geography, prodcution system, strain, and mode of operatin are not identified for commerical production, it appears that filed measurements are pre-mature. Presenter Response We appreciate the opportunity to respond to these questions. The approach was not clear partially because the project was separated into two different presentations. We have created a clarifying “Project Approach” document that is available in Yeager’s comment responses and upon request. The presentations by Sullivan and Yeager are a single project with linkages between metals, metal-containing organic toxins, human cell toxicities, and VOC production. The approach to experimental planning, which will begin with Phase II, will partially depend upon observations made in Phase I. We do not expect to find toxic concentrations of metals, but we will be looking for potential interactions between metal cofactors, algae, and pathogens that may lead to toxicity. Metals and pathogens found in field location samples will guide three types of experiments. First, the form (physical and speciation) and subsequent, cumulative uptake of metals by algae will be evaluated using the geochemical data and analysis of the algae itself. We intend to downselect metals of interest based on solution concentrations and their potential as metal cofactors in metal-containing organic toxins, as well as projected solution concentrations from systems under evaporation. Concentration, oxidation state and speciation of potential metal cofactors in toxin production will be evaluated in conjuction with Peter Moeller and his research group. Regarding human cell toxicity testing, algae is being grown in unprecedented large quantities for biofuel production. This presents several exposure unknowns for workers that are handling and processing algae on a daily basis. This project aims to address the potential occupational health hazards to workers in the algal biofuel industry. Exposure may come from direct dermal contact, or from inhalation of bioaerosols during cultivation, harvesting, or processing concentrated algae for extraction of lipids. In addition, algae are well known to accumulate metals, and may act as a vehicle for metal toxicity through these same routes. A human cell model system (HepG2 cells) was selected because it is commonly used for investigating unknown toxicity, and it is an epithelial cell type that is relevant to dermal exposure. A large panel of response end-points is being evaluated representing acute and chronic immune response, mitochondrial activity, and inflammation, among others Several of the most consistently responding endpoints will be down-selected and included in a standardized screening panel. Initially, we are testing laboratory-grown algal species that are candidates for biofuel production, beginning with Nannochloropsis salina. We will advance to samples collected at algal pond test-sites as these samples become available. In the case of the sampled sites, we will compare the responses to the responses in the “control” laboratory sites in order to distinguish responses due to the algae itself, from responses due to metal or bacterial contaminants in the industrial algal samples. With regard to the contaminants, at LANL, we are focusing primarily on identifying whether metal accumulation has a role on human cell toxicity. We are contacting as many commercial facilities as possible; however, the industry is in its infancy. We note that we rely very much on their willingness to collaborate on what can be a potentially sensitive topic. We do not believe that this is too early to assess potential hazards, particularly because growth facilities have not reached a stage of consistency of operations, nor do they exist in singular environments. The DOE has a strong obligation, as well, to support human health safety in the research operations that it has funded. Large-scale facilities that we are working with include those run by our co- PI Paul Zimba at Texas A&M, Corpus Christi, and the Texas Agrilife Station at Pecos (also a collaborator with LANL under the NAABB program). Page 169 of 223
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Reviewer Comments - EERE

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