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. Reviewer: 7 Criteria Score: 4 It appears that better definiton of commercial algae biofuel produciton systems is a critical success factor that cannot be addressed in the timeframe of this project. Presenter Response As an OHH PI (Moeller), I can assure the reviewer that there is no novel toxin work being carried out in NOAA, and apart from myself I do not believe there has been within NOAA. I mentioned earlier, each new toxin/toxic threat poses its own scientific problems. Essentially they must be treated from scratch. The efforts of the FDA, CDC and NOAA have focused on 4-5 toxins and have essentially stopped there. The vast amount of literature on these toxins only highlights the paucity of understanding algal toxin production, proliferation etc. Even these toxins (STX, PbTX, DA, CTX, Microcystins) are quite different from each other and have very little cross over in terms of scientific dealings. And these are known! Unknown toxins pose all new issues that the current literature simply does not deal with. I work with EPA, FDA, and USDA and of course within NOAA. On this topic I believe I am an expert. In phase I we will be collecting samples from 8-10 algae producers currently growing strains for biofuel production. We will also be working with several of the model organisms selected for the NAABB project (Nannochloropsis salina 1776 and Chlorella vulgaris) in Phase II and III (and other strains depending on the results of Phase I). The knowledge base concerning human health risks and algae production systems is minimal. In fact, the knowledge base is almost non-existent (at least in the public realm) for the algae currently under investigation for biofuel production. Non-toxic lab strains will inevitably behave quite differently under large scale, open culture. The body of literature regarding micro-algae culture (even in the lab) will attest to this. Pathogens are well-known to inhabit cooling towers (often full of algae), why not algae production ponds? No one has looked. This exploratory project aims to fill in some of the baseline information needed to enable any level of risk assessment. It is our hope that the information gleaned from this project can be used by professionals from other agencies (EPA, CDC, NOAA, etc.) to perform a proper risk assessment (if warranted). We are screening for known toxins (e.g. STX, PbTX, DA, CTX, Microcystins) and will pick them up quickly if they exist. However, unknown toxins prescribe a whole new set of problems, requiring new scientific methodology. Major products of this research include detection and monitoring tools developed in the course of toxin characterization. It is the first step in any novel toxin research. The second point made by the reviewer is very poignant. Our cytotoxicity assays are based primarily on the N2A and GH4C1 mammalian assays. These assays have been adopted in Moeller’s lab for novel toxin screening because, to date, they provide positive assessments for all known toxins. Irrespective of mode of action (Na+ antagonism/protagonism, Ca2+ antagonism /protagonism, phosphatase inhibition, etc) all give positive responses. After saying this, this PI agrees that our work is only as good as the assays we employ. To this end, when we observe cytotoxicity (live models) that does not set off the two aforementioned assays, we pursue other assays available to us through the Medical University of SC. Information gained from this research will be used to determine the likelihood that algae biofuel production systems (as a whole) pose a risk to human health. We intend that the tools and information obtained from this project will be used (if necessary) to help evaluate risks and form the basis for formal risk assessments in the future – once commercial algae biofuel production systems are better defined. 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. Page 164 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 Comments Reviewer: 1 This project is important yet there is little mention of how the results will be applied or shared, or what the value is. There should be interaction with EPA, but none is mentioned. Reviewer: 2 The potential for tech transfer from this project may be weak. Reviewer: 3 Results will point to framework for formal risk assessment process, including recommendations for path forward Reviewer: 4 Collaborators are involved which is good. Reviewer: 5 Technology transfer efforts (outside of the risk assessment itself) were not adequately addressed. Reviewer: 6 Too early to comment but the PI should partner with major efforts and groups. Reviewer: 7 Unless or until the algae biofuel technology comes together, risk assesment will not be ready for external communication. Presenter Response It is our hope that the information gleaned from this project can be used by professionals from other agencies (EPA, CDC, NOAA, etc.) to perform a full-scale risk assessment (if warranted). Currently DOE and NOAA are directly involved in the project and CDC will be contacted if we find evidence for persistent populations of pathogens. EPA involvement is most likely to occur if significant hazards are identified, but are not likely to be adequately addressed by federal safety regulations. Beyond information, publications, and the final report that we will produce, technologies (molecular analysis, toxin screening, toxicity assays, etc.) that are developed during this project are expected to facilitate risk assessment data collection efforts at individual facilities in the future (if warranted) and to guide the development of next generation techniques. For example, techniques to extract and analyze nucleic acids from algae samples in order to analyze all members of the microbial community will be made available. Little to no toxicity data (at least in the public realm) is known for many of the algae slated for biofuel production. In addition, even where such data does exist, the situation can and likely will change in the future for the reasons stated earlier. Major products of this research will be the technology transfer. For instance, as new toxins (or known ones from new matrices) are identified, LC, LC/MS, NMR, selective bioassays etc. methodology will be developed and subsequently adapted for toxin detection, monitoring and so forth. Even preliminary mitigation efforts can be attempted by monitoring metal speciation where this issue is involved in toxin production. Potential for tech transfer is huge and a very real product of this research. We believe our team constitutes a major group in the field. We are currently collaborating with other PIs funded through this program (Todd Lane, Sandia). CDC will be contacted if we find strong evidence for the sustained presence of a human pathogen in any of the production system samples that are examined. Page 165 of 223
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Reviewer Comments - EERE

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