Reviewer Comments - EERE

2011 Biomass Program Algae Platform Peer 

Review: Review Comments 

Table of Contents 

Algae R&D Platform Overview ................................................................................................................ 2 

Project: 9.5.1.1 Los Alamos National Laboratory ..................................................................................... 9 

Project: 9.5.1.4 Cellana ............................................................................................................................ 19 

Project: 9.5.1.5 Arizona State University ................................................................................................ 26 

Project: 9.5.1.6 CABComm ..................................................................................................................... 33 

Project: 7.2.1.7 University of Nebraska Lincoln ..................................................................................... 38 

Project: 9.6.5.3 NREL ............................................................................................................................. 44 

Project: 9.6.5.2 Argonne National Lab .................................................................................................... 50 

Project: 9.2.2.2 University of California - San Diego ............................................................................. 55 

Project: 9.2.1.2 University of Georgia ..................................................................................................... 65 

Project: 9.2.2.1 University of Maine ....................................................................................................... 71 

Project: 9.2.1.1 Montana State University ............................................................................................... 77 

Project: 9.6.1.3 Pacific Northwest National Laboratory .......................................................................... 83 

Project: 9.6.1.2 PNNL ............................................................................................................................. 91 

Project: 9.1.3.1 INL ............................................................................................................................... 100 

Project: 9.6.1.6 PNNL ........................................................................................................................... 109 

Project: 9.6.1.1 National Renewable Energy Lab .................................................................................. 117 

Project: 9.6.5.1 Sandia National Labs.................................................................................................... 122 

Project: 9.1.2.1 Brookhaven National Laboratory ................................................................................. 127 

Project: 9.1.3.2 INL ............................................................................................................................... 135 

Project: 9.2.2.3 National Renewable Energy Laboratory ...................................................................... 142 

Project: 9.1.2.2 Sandia National Laboratories ....................................................................................... 149 

Project: 9.6.1.5 SRNL ............................................................................................................................ 158 

Project: 9.6.1.7 Los Alamos National Laboratory ................................................................................. 167 

Project: 7.9.2.2 Desert Research Institute .............................................................................................. 176 

Project: 7.7.2.18 Brooklyn College ....................................................................................................... 185 

Project: 7.9.1.4 Cold Spring Harbor Laboratory ................................................................................... 193 

Project: 7.9.2.1 Old Dominion University ............................................................................................. 199 

Project: 9.6.6.3 PNNL ........................................................................................................................... 204 

Project: 9.6.6.2 Sandia National Labs.................................................................................................... 212 

Project: 9.6.6.1 NREL ........................................................................................................................... 217 

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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. 

Algae R&D Platform Overview 

Title: Platform Overview - Joyce Yang 

Presenter: Technology Manager 

Presentation Date: Thursday, April 07, 2011 

Criteria Avg. Score Stud Deviation Count 

Relevance 7.67 1.60 6 

Approach 7.00 1.83 6 

Progress 6.50 2.36 6 

1. Relevance 

Please evaluate the degree to which: 

Platform goals, technical targets, and barriers are clearly articulated and logical 

Platform goals and planned activities support the goals and objectives outlined in the MYPP 

Achieving Platform goals will increase the commercial viability of biofuels. 

How could the Platform change to better support the Biomass Program goals? 

Reviewer Comments 

Reviewer: 1 Criteria Score: 5 

The very high level OBP goals as targeted by EISA were spelled out. There has been significant 

investment in projects over the past two years, including one massive investment in NAABB. But it is 

clear from the presentation that goals are yet to be set. There is an implication from one of the slides that 

Sustainability and Analysis were done to an extent up front. I am not sure how apparent that is in the 

project selection, however. Many of the projects seem to suffer from a lack of analysis-based goal and 

deliverable setting. An improved delivery here might discuss some back-of-the-envelope technoeconomic 

assessments which allow one to key in at early stages on specific areas of the slide titled 

Roadmap Categories, further expounded in the next 4 slides on next steps and integration. In other words, 

a little deeper technical description would have helped make a better case for project selection. One could 

even have pasted the specific projects on the Integration diagrams. There are few enough programs that 

this could have been done and maybe have bolstered project selection decisions. 

I unfortunately came away with questions as to how certain projects were selected. Some of the projects 

lacked a mission-focus that I would expect for projects which are basically determining whether algae 

should be a feedstock or not. If FY2013 is the year of "Downselection" then each of the Projects should 

concretely map onto the pathway leading to that selection, and goals/deliverables should have been 

developed which will allow that to happen. This should have been bolstered by at least rudimentary 

techno-economics. Maybe the projects were selected this way. It sure was not apparent from some of the 

presentations. 


The general platform goals involving feedstock development, cultivation, harvest, extraction and 

conversion, and fuel production/distribution are sound. It is keenly important to connect basic and applied 

scientists and to make sure each end of this equation understands the potential and limitations of the status 

quo across the platform. 


Effort by the program is directly along the lines of the Algal Biomass mission 


Although the Algal Biofuels Roadmap was rather uncritical and did not do much to narrow down which 

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technologies are suitable for biofuel production, it has provided a good framework for organizing the 

research program. Thus, the objectives of the funded projects are in-line with what is needed to resolve 

the many questions on algae biofuel production. 


Good presentation of Program goals and objectives. 



It is important to recognize these research activities as a long term effort with low probability of success, 

with the potential for very high benefits. The focus on productivity, harvesting, de-watering, extraction 

and process economics is highly relevant. Algal biofuels projects for specific geographies do not support 

platform goals and technical targets because the geographies have not been chosen based on science. 

Some of the detailed work on co-products, upgrading oil to fuel, and risk assessment appears to be 

premature because a neither commercial strains nor commercial processes have been identified. 

2. Approach 


Platform approaches are effective, as demonstrated by the extent to which: Platform milestones and 

organization; project portfolio; and strategic directions facilitate reaching Program Performance Goals as 

outlined in the MYPP 

The Platform portfolio is focused and balanced to achieve Biomass Program and Platform goals, as 

demonstrated by: Work Breakdown Structure; unit operations; and pathway prioritization Please explain 

your score by commenting on the strengths and weakness evaluated 

What changes would increase the effectiveness of the Platform? 



There was a framework established for constructing the evaluation of algae as a potential feedstock for 

biofuels production in the future. As mentioned previously, there did not appear to be significant evidence 

of direction-setting using technoeconomic assessments. The portfolio should then be balanced with 

respect to the key drivers. It may be that most of the funds should be spent on biology, since without 

adequate productivity per organism, algae will not be a cost-viable feedstock. One would like to hit this 

hard so as to be able to come to a decision point in FY2013 or FY2014. Part of the OBP program should 

be a rapid identification of those feedstocks that are viable and those that are not, so as to be able to 

throttle expenditures here and spend more on the next economic bottlenecks. 

As a general comment, some of the programs are doing a better job at the WBS to provide useful 

deliverables, while others would seem to require some oversight. 


The type of review meeting held here should be useful to the personnel of the projects. Some of the 

projects have great focus by dedicated and expert scientists/engineers. However, there are a number of 

weak projects in the current portfolio. It is especially important that modelling projects use realistic input 

data and have excellent knowledge of background literature. In some cases, the goals are appropriate to a 

project but the likelihood of success by that group of investigators is low due to mismatched expertise. 

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A variety of complementary approaches are being pursued, which increases chances of success. 


The use of consortia is essential for coordinating research on the many steps in the algae biofuels 

production chain. This approach was a good choice, but flexibility should be maintained, allowing new 

participants and technologies to enter/exit existing consortia where appropriate and useful, as determined 

by the PIs and the DOE managers. The existing practice of at least weekly contact of DOE managers with 

the major consortia should be helpful in achieving good results. Although time-consuming for the 

managers, such coordination should facilitate the continual improvement and adjustment of the research 

plan. The DOE managers should assist in the rapid culling of technologies from a consortium that are not 

likely to meet economic, energy balance, or LCA criteria. The PIs may have difficulty severing 

relationships with collaborators without some outside impetus, although there is no evidence of that 

problem thus far. A general weakness of many of the project presentations was a lack of well-defined 

technology down-select criteria nor any a priori quantitative estimates of technology performance in 

terms of cost, energy balance, and LCA. While the national labs are generating open-source TEA and 

LCA software, good initial assessments should be done by the project proponents at the proposal phase, 

then to be updated at least annually during the project. These assessments should be gaged against 

performance criteria set by the OBP algae managers. If fact, these suggestions may have already been 

implemented at the proposal phase, but then not emphasized in the presentation template used for the 

review. Related to the above discussion, the algae program should fund both basic and applied research, 

with the majority being applied. Each project should strive to define to what extent the research is basic or 

applied. This would aid particular reviewers who are prone to prefer applied work to give an allowance 

for the apparent lack of immediate practicality of some of the projects. However, the main value in better 

defining basic/applied would be to prevent basic science projects being presented as applied, which 

comes across poorly to reviewers. Many of the individual projects, not consortia, are also valuable. Of 

course, independent approaches are needed, perhaps funded on a seed basis, to be incorporated into 

consortia as results warrant. The algae program managers should work to coordinate research funding and 

RFPs with ARPA-e, the Office of Science, NETL, and the USDA. The OBP algae program should be 

involved in all DOE algae work because the algae program managers have already become expert in the 

topic and duplication of expertise and uncoordinated research funding is undesirable. Of course, the rapid 

rise and fall of funding for the algae program is detrimental to sustaining the research groups who will 

bring algae biofuel to fruition over the long-run. Algae biofuel development will need to be a well-funded 

on-going effort even if early success is found. The implementation, maintenance, and improvement of 

productivity on tens of thousands of hectares of algae production will require on-going effort akin to the 

agricultural research funded by USDA. Inclusion of non-fuel co-products as valid research topics in the 

algae program should be continued. Of course, the link to biofuel production must still be present. One 

exception to that requirement would be if the algae technology has promise for leading to significant 

savings in fossil fuel consumption or greenhouse gas emissions. These two benefits should be considered 

nearly as valuable as actual transportation fuel production. Coming to a smaller detail, the proposal 

review process may need to be improved based on some of the marginal projects presenting during the 

review. The small pool of algae production experts to choose from could be the source of the problem. 

The OBP should continue to expand their list of potential reviewers. Members of the Aquatic Species 

Program are good candidates since they have longer experience and more expertise than many in the field 

today. The panel used in this program review had a good diversity of expertise and experience. Genetic 

engineering expertise is important to have in algae reviews. 


Good presentation of Platform approaches and portfolio. 

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The approach of having one large, highly funded consortium working in parallel with two smaller 

consortia and multiple academic and private projects is likely to produce redundancies. Bringing the 

dispersed efforts into focus on the key technical hurdles is daunting. 

3. Progress 

Please evaluate the degree to the Platform is progressing towards achieving Biomass Program and 

Platform goals, specifically in reference to: meeting performance targets and likelihood of achieving the 

goals presented. 

Please provide recommendations for improvements for tracking progress. 



Based on the individual presentations, it does not appear likely that the platform goals will be met. I think 

the OBP should be hardening these goals/milestones/deliverables for the total platform and for each of the 

individual pieces. I am not speaking about making the FY2013 technology selections now, I am saying 

that it does not appear that the component deliverables to allow the FY2013 selection have been 

formulated with enough clarity and detail to maximize the likelihood of selecting in FY2013. With these 

deliverables in hand, meet with the various projects to map their contributions onto this same deliverable 

timeline. Then attempt to build a fire. I presume since funds have been spent totally, and unless these 

timelines and deliverables were built into the contracts originally, then there is little recourse except to 

convince the project leaders of the value of delivering in line with the Platform. Again these need to be 

hard-wired and not general deliverables. 


The largest project has excellent personnel and multiple scales; it is the place where there is the most 

promise of success. It is essential that this project doesn't get too side-tracked by "neat stuff"; basic and 

applied scientists and engineers in this project need to know on a monthly basis what each group has 

found that advances the project. 

Some of the "sampling from the wild" for feedstock improvement is a little too much like a fishing 

expedition; little hypothesis-testing was evident in sampling location selection. Many models appear 

completely unrealistic (poor research of input data, lack of ground-truthing and consideration of multiuses 

of land area, sea level rise projections affecting coastal projects, etc.). In general, too many projects 

exhibited a failure to know the background literature related to their project goals. 

The best targets are likely to be non-GMO species because open systems are too subject to bird dispersal. 

Thus, the "neat stuff" related to transformation should be evenly balanced by traditional strain selection. 


Too early to judge, as this is just the beginning of this program. Nevertheless, this first-time peer review 

process showed a vibrant group on investigators and truly dedicated program staff. 


The creation of the Roadmap and the MYPP early in the program was important. Most of the projects 

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funded recently have value and are making progress. This program review is also a sign of progress and 

organization. 


It is too early in the process to evaluate progress in the algal biomass program since many of the projects 

have just begun. 



Expectations for progress need to be calibrated to the vastness of the challenges in commercializing algal 

biofuels. Most of the projects reviewed had less than one year of activity. Identifying technologies to 

evaluate and getting a techno-economic model in place to use as a tool to determine viability is good 

progress. 

4. Overall Impressions 

Please provide an overall evaluation of the platform, including strengths, weaknesses, and any gaps in the 

Platform portfolio. 


Reviewer: 1 

The view and assessment of the algae platform are very intriguing. It is like starting from preliminary 

data, and then trying to move to commercial reality in a fixed timeline of 10 years. If one subtracts from 

the 2022 target a typical and sufficient length of time for development and demonstration of all the 

downstream tasks leading to biofuels, one is left with the need to come to conclusions on the viability of 

algae as a viable feedstock possibility in a very short timeline. Some of the projects will contribute to that 

decision point, while others seem not likely to do so. A reformulation of platform 

goals/deliverables/timeframe may help the OBP personnel dialog and bring the PIs along so that the OBP 

may meet its 2022 goals. Overall it is easy to come to the conclusion that people (PIs, scientists, PMs, 

DOE, government officials) have already concluded either that 10 years is still so far away or algae is not 

really in the 2022 picture, so why push? If this lack of drive to meet 2022 goals is real, and algae is the 

advanced, advanced feedstock for 2040 fuels, then re-trenching to the really basic drivers at far less 

expenditure would be a better move in terms of overall OBP goals. If people think there is still plenty of 

time in the 2022 window to spend years on determining algae as feedstock possibility, they should also 

look at some realistic deliverables and experiences in the other unit operations. Overall, OBP should set 

tighter operating specs for results delivery and move deliberately and rapidly. 


I think stronger project selection would be particularly helpful to attaining goals. No competition should 

be limited to one entity (i.e., national labs vs small business vs academia). If DoE can forge strong people 

from business, engineering and basic science into a project, that is likely to have the most success. The 

largest project is an example of this, and with tight coordination and internal communication, it should 

succeed because of its funding level (large!) and personnel. A number of other projects are strong, and 

some others should be eliminated. 

Much stronger peer review is required at DoE before favorable funding decisions are reached. A revised 

peer review process might have individual reviewr comments/scores sent only to PIs, because this permits 

reviewers to provide detailed and constructively critical assessments of the project to help PIs. For public 

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distribution, a panel summary could be released. This is the closer to the NSF procedure, which has an 

excellent track record in terms of scientific result and taxpayer benefit. 


Strengths: Enthusiasm of the program staff and team; clarity of presentation and clarity of vision for the 

program. 


The OBP algae program team all seem very capable and dedicated to developing the sound information 

from the research that is needed to guide DOE decisions. All in all, I was heartened by the straightforward 

approach and open attitude I sensed in the program team. I am much more confident in the OBP 

having seen this team interact during the review process. The algae program is new but making laudable 

progress, especially in light of the management burden that the sudden influx of ARRA and 

Congressional funding must cause. I am confident that if DOE can continue to develop the expertise of 

the algae program managers that the research projects will reach a level of value that is exceptional 

among DOE programs. On the other hand, management turn-over and uncoordinated algae efforts across 

DOE will obvious give a poorer overall result. 


The platform approach is good, but a considerable number of the projects seemed unnecessarily weak 

(even at this early stage of review). 



The greatest strengths of the program are the definition of key hurdles to commericialiization, the 

identification and funding of individuals and institutions with the ability to address these hurdles, and the 

implementation of modeling tools to evaluate progress. A weakness is the ability to define specific, 

relevant metrics to track all of the individual projects and the overall effort, and to prevent redundant 

work. 

There is a great weakness in the projects that are forced into a specific geography. Putting challenging 

geographical constraints onto the extremely difficult commercialization of algal biofuels virtually ensures 

failure of those projects. 

Uneven long term funding with high spending for 3 years also creates significant program weaknesses. 

Initial funding should be at a level that matches the effort necessary to demonstrate feasibility with the 

highest existing level of talent. If key hurdles are addressed and potential pathways are defined, funding 

should increase over time, not drop precipitously. 

5. Additional Recommendations, Comments, and Observations 



It is clear that accountability of the investigators is an issue for these projects. Either an installment 

contract needs to be devised with deliverable attainment a requisite for further funds, or the deliverables 

need hard-nosed formulation before the contract is awarded. OBP seems to have its mission firmly in 

front, but there is not the same devotion from all of the awardees in my perception. The appearances are 

that some are just in it for the funds, or to extend a research program, or to peddle a favorite piece of 

technology, or for the politics. A very tough management and execution challenge! 

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This is an important area for DoE; some biofuels must come from algae (due to the chemical realities of 

different molecules for different purposes). Also, there are additional products (plastics) that will demand 

algal biofuels, even when we kick the petroleum habit. If algal biomass production were coupled to 

watershed clean-up (e.g., the Mississippi, the amount of biomass produced could be significant and drive 

a much larger part of total biofuels in the future from algal sources. 


There is an absolute need of continuity in this Algal Biomass program. Three years of R&D is not 

sufficient to bring about a successful outcome, as the case would be with any and all new and ambitious 

programs. The stakes for the country and the world are enormous, and the effort ought to reflect this very 

fact. I should emphasize that microalgal biofuels is an area the private sector has embraced with 

enthusiasm, and where private industry funds have been invested. The federal government and the DOE 

ought to take a queue from the trail of "private sector investments" in this field and to help accelerate this 

line of R&D with continuous support beyond the initial three-year period of funding. 


There are no additional recommendations. 


For the large projects, it is vital that the managers pressure the investigators to objectively evaluate and 

integrate their individual results relative to large scale production. Otherwise, it will be too easy for 

individuals to focus on individual processes and be left with an array of unrelated results. 


It would be helpful to have independent experts develop the road map, as opposed to those who will 

shortly be applying for large expenditures of funds along the lines of the roadmap. 

Also, it would be helpful to permit a longer time for preperation from the time of the RFP, and to instill a 

more rigorous review process of the proposals. 


It might be useful to require projects to establish specific, quantitative goals direclty related to algal 

biofuel commercial requirements. 

Focus on the biggest technical hurdles--growth, harvest, and extraction. Some of the risk assesment and 

co-product studies appear to be premature because strain and process will dramatically affect 

characteristics. Co-product and risk assesment work should be limited to early feasibility demonstration. 

The program will not fail due to lack of technology to convert lipids to fuels, so this work should be 

minimized. 

Peer Review Presentations need more structure imposed by DOE--a summary slide of specific 

quantitative goals, baseline, achieved, and expected results against those goals, and decision points based 

on resutls. Any economic projections need to be accompanied by assumptions and calculations. 

There are many strain screening efforts. A centralized database for the results might be valuable. 

Validation of screening processes against strain performance in pilot operations is needed. A 

standaridzied, small scale, rapid measuremetn system for strain performance validated against longe term 

outdoor cutlivation would be extremely valuable. A statistical analysis of algae biodiversity based on 

strain data already collected might be a useful tool to determine if continued bioprospecting is likley to 

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find a strain with traits sufficient for commericialization. 

Process research efforts appear to be hindered by a lack of algae supply. Considerable resources are used 

and time delays incurred by projects growing algae on very small scale. The DOE should consider 

developing algae supply agreements with facilities that can supply kg to 10's of kg dry basis quantities of 

custom grown algae feedstock to researchers. 

NREL techno economic modeling is an essential tool and should be expanded to evaluate scenarios based 

on pilto results and conceptual projections. 

Project: 9.5.1.1 Los Alamos National Laboratory 

Title: NAABB An Algal Biofuels Consortium 

Presenter: Jose Olivares 


Criteria Avg Score Std Deviation Count 

Approach 5.71 1.28 7 

Progress 5.57 0.49 7 


Critical Success Factors 5.71 1.16 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. 



This is an enormous project There has been significant effort at lining up the approaches on several 

different lines of attack. 


Scaled well from lab bench to large facility at Pecos. Project participants are strong. However, all 

participants need to focus on the end goal of biofuel production, and it would likely be helpful for a 

project meeting to be held soon at Pecos, to highlight the importance (and challenges) of scale. 


The NAABB Algal Biofuels Consortium aims to address key barriers across the entire chain of algal 

biofuels production. As such, it is an integrated program that aims to develop tools via which to address 

current issues in algal biofuels. 


See Overall Impression text. 


Very broad goals encompass almost every topic within biofuel production. 

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Altough the presentation was professional, and the individual topics (unit operations) discussed of 

obvious quality, it was very difficult to see how this approach improves upon the most recent reports of 

Benneman and co-authors It appears to be a strong collection of individual researchers, each with their 

own expertise, collected from labs around the U.S., performing their own research. Although the PI layed 

out a competant set of "criteria" or objectives that has to be met to achieve success (slide 2), the follow on 

slides simply looked like individual PI's sent along slides from their group and the PI massaged them 

together. This reviewer did not see a clear system of unit operations put in place up front, with specific 

objectives (and input and outputs for all) laid out, that the "team" had to develop. Until this is done, until 

the PI steps up and states "this is how we are going forward and these are the targets per unit operation" 

this consortium can be limited by a culture of self-preservation that undermines the down select process. 

The PI is strongly encouraged to read the reports of Benneman and co-authors and to understand how 

most of the restrictions stated in those reports will dominate, and how he will use this "experience" and 

"common sense" to rapidly scale this program back to a fundamental, agriculture/farmer based approach. 

Rapid downselection with an aggressive application of common sense should be applied rapidly before 

this program becomes too difficult to manage towards large scale commercial deployment. 


Program has well-defined, specific, and relevant goals with quantitative values for productivity and costs. 

Milestones and decision points are identified on a time scale. Activities that should be done in sequence 

are being driven in parallel, possibly because of the short term nature of the funding. 

Presenter Response 

As the FOA for our proposal stated, the main NAABB program goals are the development of 

technologies for biofuels development. The FOA also required that our program add components 

necessary to develop all aspects of biomass generation and utilization, in order to increase the economic 

value of the overall biofuels development, such as would be implemented in a biorefinery system. 

Therefore, the bulk of our program leads to biofuels development from algal lipids and biomass, along 

with power generation, chemical and agricultural coproducts developed from lipid extracted biomass. We 

have had several face-to-face project meetings and many others, in which scale-up challenges have been a 

major topic. Similarly, our management team and several set of investigators have travelled and met at 

Pecos and other production sites to review the process and understand the challenges. We will continue to 

do this and will take into consideration having a future overall program meeting at one of our large-scale 

facilities. The NAABB program uses the knowledge gathered in the Acquatic Species Program (ASP), by 

John Benemann and others, as a starting point. For example, we utilize Nannochloropsis salina as one of 

our main production strains. Unfortunately, many of the other strains identified by the ASP have been 

lost. Therefore, the NAABB prospecting efforts is helping continue to identify potential useful strains for 

cultivation. We also realize the technological advances that have occurred since the end of the ASP, for 

example, genomic and transcriptome sequencing capabilities have made it much easier to identify target 

genes for regulation and modification. Thus, the NAABB program is utilizing new high throughput 

sequencing and proteomics capabilities to advance our knowledge of algae and develop successful 

productive strains of algae. We are examining innovative ways to increase lipid production. This includes 

the identification of genes via systems biology and random mutagenesis approaches and developing tools 

to transform likely production strains with those candidate genes that prove helpful. Crop protection also 

falls under this area of increased lipid production by potentially decreasing down time due to crop failure, 

i.e., pond crashes. The FOA for this program required the formation of a consortium with a broad 

representation from academia, industry and national laboratories. In forming our consortium we chose 

institutions that brought capabilities that were complimentary and appropriate across the developmental 

process for biofuels. We have assembled a team that has unique capabilities, yet is willing to work 

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together to develop the NAABB objectives and goals. The NAABB program has over 70 individual task 

and subtask areas, within a very structured work break down schedule with milestones, deliverables, and 

decision points. These are contained within our Statement of Project Objectives and our Project 

Management Plan, as submitted and negotiated with DOE. A 45 minute presentation, at such a level of 

detail, would not have been possible. Our program has incorporated a number of check points, 

assessments, and decision points for the processes and technologies being developed, the first of which 

are being implemented at months 12, 15, 18 and 24, in our program. These will generate a downselection, 

rescoping, and/or refocusing for many of our processes to achieve timely changes in R&D to 

achieve our goals and objectives. Our goal is to make production of microalgae a farming endeavor that 

is: economically feasible, energetically positive, and environmentally sound. This goal cannot happen 

without new scientific and technological breakthroughs. And, certainly it cannot happen without the 

underlying foundation and infrastructure to support such a farming endeavor, e.g., seed (algal strain) 

development, sound cultivation practices, economical harvesting, and process logistics. All of which are 

major components of the NAABB program. 

2. Technical Progress and Accomplishments 

Please evaluate the degree to which the project has made progress in its objectives and stated project 

management plan has met its objectives in achieving milestones and overcoming technical barriers. 



Progress has been made even though the project has not been in place for very long. 


Development of ARID raceway, genomic information, and strain improvement work is promising. 

Acoustic focusing is promising in recovery. GREET analysis for life cycle analysis is important. 

Recycling of N and P after lipid extraction will be critical. Bioproducts of fuel include aquaculture feeds, 

which is very promising but what is the omega 3 FA content after lipid extraction, if any? Without this, 

the feed quality will be much less valuable. 

Concerns/monitoring of competitors and pathogens in open pond raceways is more transparent than many 

similar efforts; chemical and engineered disease resistance is proposed; I think it is unlikely that GMO 

algae will be approved in open pond raceways due to wide dispersal potential of such algae into the wild 

at 100s of km from sites. Strain selection needs more attention for disease resistance. 


This is the first year of operation of the NAABB and progress is modest. However, this is to be expected 

as all projects have a long ramp-up time in the beginning. Question: How does NAABB plan to monitor 

and assess progress by individual participants? I ask them to develop a quality control mechanism that 

would be applied to all participants. As one criterion, I would recommend simple Gantt charts for each 

and every of the NAABB projects, and application of the criterion of "peer-reviewed" publications, as a 

benchmark. 




Unfortunately, there was not enough detail for evaluation. 

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Score: 5 

It was a competant presentation of work presented, from each group, but very little of it was obviously 

linked to an overall goal of developing a single commercial production system. It was not clear if any 

single unit operation was aware of what the upstream unit operation was going to deliver and what the 

downstream unit operation was going to need to receive. One should be careful touting numbers like 

0.11$ per gallon harvested (in the extraction section). Groups are always trying to insert cost instead of 

energy ratios, at each unit operation. It may sound great to have 0.11$ per gallon harvested, but if one 

process is processing streams that are 99.9% water, this is likely not such a low number when one 

considers the total amount of gallons to be processed. One should show how much energy (or power) is 

consumed per unit of fuel (or volumetric flowrate) processed. This ratio should be less than 10% of the 

overall system for extraction, and 15% for dewatering and harvesting (or 25% if combined into one step). 

It’s important that the use of cost is replaced by energy use in systems analysis. If the amount of energy 

used is less than that produced, the cost balance will sort it self out. But if more energy is consumed than 

produced, the cost balance will not sort itself out unless free sources of energy are leveraged into the 

system. 

One should not just state low energy input for the acoustic harvester. Low relative to WHAT? One should 

show the power input required per volumetric flow of fuel processed (and use this to develop an energy 

ratio). It is conerning to see respective extraction (or harvesting) devices touted without a proper metric 

for the energy consumed per unit energy produced. The project should be specific about the use of energy 

balances that track the unit energy consumed per unit energy produced. 


Progress in Algal biology and cultivation are co-dependent and are not quantifiable until cultivation data 

is available, but the approach and technical expertise is a positive. There appears to be good progress in 

identification and preliminary testing of several novel technologies for harvesting and extraction. Because 

biology, cultivation, harvesting, and extraction will all have significant impact on oil and coproducts, 

much of the work on fuel conversion and feeding studies appears to be premature. 


As presented in the Peer Review, the NAABB program for strain selection flows into a cultivation 

pipeline that tests these strains in outdoor environments. This will provide screening for strain robustness 

against environmental factors such as predation, bacterial infections, etc. Our program also incorporates 

the development of technologies that will help provide resistance from such infections, through the 

addition of bacterial controlling substances and breakdown of biofilms produced by such organisms. 

Disease resistance is a good parameter to test and one we certainly keep in mind. Our initial selection is 

on growth and lipid productivity but we intend to put the best strains into a cultivation environment. 

NAABB is already monitoring progress in a number of ways, including: site visits by our project 

management team, monthly reports, financial and fiduciary reporting, face-to-face status reviews, 

monthly team meetings, bi-weekly meetings with team leads, and weekly meetings our operations, 

executive team and DOE. As our supporting slides show, the NAABB investigators have already credited 

the NAABB program with over 20 manuscripts in print or under review in peer reviewed journals. We 

have also developed a website with regular highlights. We have an obligation to protect intellectual 

property, make it available first to our partners for licensing opportunities. Once this obligation is met, 

any IP not licensed is then made available to the general public through the innovating institution. 

Further, NAABB investigators have made over 100 presentations at national and international 

conferences. Our investigators have and are coordinating technical sessions at national conferences. 

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Similarly, we are chairing and organizing an international conference in St. Louis later this year, in which 

many NAABB investigators will be presenting their work along with peers from the international 

community. 

We are in total agreement with the reviewer comments. For all of our harvesting and extraction processes 

we are developing a process flow model that determines all of the inputs and outputs both for mass 

balance and energy consumption and/or generation. These can then be converted to a cost analysis. The 

preliminary cost results presented for the LANL acoustic harvester represented the results from such a 

detailed analysis. 

3. Project Relevance 


The project both identifies with and contributes to meeting the platform goals and objectives of the 

Biomass Program Multi-Year Program Plan 

The project has considered applications of the expected outputs 



If the consortium can perform as a team, the relevance will be up there in scores. The potential is there. 


Highly relevant and promising. Costing of all stages needs strong attention in year ahead. 


Project is well aligned with the Algae Biomass MYPP and roadmap, and efforts are in direct support of 

this EERE program. 




The stated goals are closely aligned with the platform goals. 


Score 7: The presentation was very good in terms of presenting its talk under the umbrella of the Biomass 

Program's objectives, and the Program's output. 


NAABB activities are a close reflection of the Biomass Mulit-Year Program Plan. 


The magnitude of the NAABB program makes it very challenging to put together a presentation for this 

type of review process. Therefore we strived at a 45-minute presentation that provides an overview of the 

NAABB framework, innovation, status, results, and strategy for the future. In most cases, when we 

presented innovation or results, we were faced with only the time to present a bullet, if at all presented. In 

some cases, we chose to provide a particular highlight of the scientific innovation and results from our 

program. 

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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) 



These CSFs have been stated in the presentation. They also are tied to each thrust. 


Genomics' concentration must be demonstrated to be worthwhile at intensity developed. 

Disease/competitor problems must be overcome; stochastic events that cause crashes must be modelled 

realistically and costed. 

Probiotic bacterial angle likely to be significant. 

The most important thing now is to show cost-effective production of algal lipids to methane to 

bioproducts. Scale concerns must be met. 


The project has systematically identified all possible factors and barriers that need be addressed for a 

successful outcome. 




Unfortunately, there was not enough detail for evaluation. 


Please evaluate the degree to which: 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 Other than slide 2, not much in this area. The project has presented adequate plans to 

recognize, address, and overcome these factors. This was a weakness to the presentation as little 

discussion or mechanism by which this would be approached was presented. 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) 




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end-use) 

iv. Environmental sustainability (example: water use, GMOs, energy consumption) The PI consistently 

showed Sustianability as a large "arrow" at the bottom of the slide that suggested it was to be the overall 

driver. However, the driving criteria for sustainability and how they would be imposed for the 

downselection was not apparant. Then again, to proper evaluate a consortium of this size, a full day 

should have been given to their presentation. 


Success factors have been clearly recognized. At the current state of technology definitive plans to 

overcome and address these factors can not be defined. Plans appear to be appropriately focused on 

determining the feasibility of potential solutions that might be contributed to a viable biofuel pathway. 

Successful demonstration of feasibilities will provide an opportunity to impact commmercial viability. 

More emphasis is needed on developing ameasurement system to quantify strain productivity which is 

validated against long term outdoor cultivation. 


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. 



This project will require an extraordinary level of coordination to deliver the outcomes. There are no 

apparent public strategies or demonstration strategies that would be valuable. The use of iPhone is open to 

question. There are already journal publications. 


Tech transfer of raceway design, acoustic focussing etc. is strong. Pubs and presentations look good. 

Critical year ahead to demonstrate cost-effectiveness of biofuel production at scale required. 


Too soon to tell. 




The project appears to be highly coordinated, but the project is too large (and presentation too short) for 

adequate review of this topic. 


This was quite extensive but might actually be the Achilles heel of this program. The collaborations are 

so extensive, and the expectations of large scale microalgae production for fuels so overstated, the PI 

might find it difficult to down select to the final process. There is concerne that the number of 

collaborations is too big for a single PI to manage. 

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Internal sharing appears to be strong. Broad outreach in economic modeling is commendable. 

Development and broad sharing of an algae biology data base with key properties of various strains might 

have value. 


Direct response to the Tech Transfer Section: As shown in the peer review presentation, our program 

structure provides for a thorough and very supportive management team that manages the day-to-day 

activities of NAABB. This includes three technical directors, 13 technical team leads, 2 operations 

managers, a full time project manager, an industry and intellectual property manager, and the full 

financial and subcontracting engine at Donald Danforth Plant Science Center. Without this structure, I 

would agree with the reviewer. 

While there are no more responses to the Tech Transfer section, with the 4,000 character limitation in 

each response section, there are some additional comments referring to the Overall Impressions section: 

We want to thank the DOE for the opportunity to respond to the Peer Review Committee comments. We 

found that the comments reflected some very positive views that the NAABB program is making an 

impact, even this early on in the performance period. The NAABB primary focus remains biofuels 

production. Therefore, if upstream processes extract or degrade the fatty acids left in the animal feed, this 

will affect its nutritional value. Thus, NAABB investigators are trying to determine the value of such 

feed. Obviously, there is a feedback loop, and if such high value components can be extracted and/or left 

in to add additional value to the feeds we will incorporate such processes. Delivering feed to an animal in 

a format that it will be consumed effectively is key to this industry. It increases handling characteristics. 

Not all potential feed materials pelletize easily (pelletizing distillers grains is currently a challenge for this 

industry) and in some cases additives need to be introduced in order to achieve practical formats. 

Achieving this can increase the value of feed by as much as $112 /ton. In the time allotted all we could 

present were major goals and objectives, e.g., in slide two of the presentation. Nevertheless, the NAABB 

program does incorporate understanding net energy production through life cycle assessments on all 

processes that are carried forward. A comparison of these net energy production processes can then be 

available to policy makers and industry for further decision making on viability of any one process. 


Please provide an overall evaluation of the project, including strengths, weaknesses, the project approach, 

scope, and any other overall comments. 



This project hinges on many parties keeping their thrusts on target. Some of the tasks could use more 

injection of industrial reviews and maybe even technology contributions. 


The team is good and seems well managed. The level of funding is extremely high and thus expectations 

of the project managers for real biofuel scale production should be equally high. Balancing "neat" 

science, much of it required for success, with pragmatic advances is one of the challenges to the project. 

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Strengths: an impressive array of projects and expertise is brought together in this NAABB effort. 

Challenges: The ability to successfully oversee and coordinate all project-components. How do they plan 

to monitor and assess progress by individual participants? ARe there metrics and milestones for each part 

of the NAAB effort? How do they plan to disseminate information and the know-how resulting from this 

effort? I recommend the criterion of "peer-reviewed" publications and Gantt charts, as benchmarks. Does 

the consortium plan to hold regular meetings, where all participants would present their progress? The 

DNA sequencing effort of Botryococcus braunii was initiated by a group of interested (Botryococcus 

community) scientists and actually started by the JGI well ahead of the NAABB; it is inappropriate for 

the NAABB to claim this as one of their accomplishments. 


Overall Impression: 5 

The project abstract correctly identified the basic key technical challenges: "1) Algal strains that can be 

cultivated in real-world conditions and harvested with minimal energy; 2) Technologies that are scaleable 

and provide the energy return on investment; 3) Technology integration with needed nutrient, water, and 

other recycles; and 4) Sustainable technologies with respect to environment, cost and permitting." The 

presentation identified cost goals, such as $2.10/gal oil, presumably a total cost. Quantitative goals on net 

energy production and LCA issues were missing. 

A major lack in the presentation was evidence that the proponent have made quantitative projections on 

whether their suite of candidate technologies has the potential to meet the key technical challenges and 

the cost goals. Target productivities, oil contents, etc. were given, so management should demonstrate on 

an on-going basis that these technical targets would actually lead to the cost target. Do both the 

productivity and lipid content have to be near the maximum conceivably feasible to reach $2/gal? How 

many major breakthroughs are needed to reach this goal? Then results/progress from the technologies 

could be measured against the needed performance. In particular, somewhat reliable projections for some 

technologies (eg, ARID cultivation system, harvesting, and extraction technologies) should be possible 

immediately. Of course a down-select is scheduled, but the methodology of that process remained 

obscure. 

The strain selection process does not seem to be designed well. The slides state that either >300 or >500 

stains, mostly collected from the environment, have already been screened. Why would any of these 

survive under cultivation conditions? The lab culturing conditions described do not recreate the main 

stressors of outdoor cultivation. The current methods seem to be brute-force only, not efficient, and not 

well thought out. 

The good aspects of the management plan are the frequent PI meetings and reports to DOE, but still 

needed is the better defined goal-vs-performance assessment mentioned above. The outdoor productions 

systems are a great strength of the program for demonstrating a tangible result. It is good that production 

strains are currently being tested outdoors in addition to the less promising lab strain selection procedure. 

Other areas need strengthening: The purpose of Solix PBR effort was not explained. The wastewater and 

produced water efforts were not well described. The animal feeding trials will require large quantities of 

biomass and presumably several years of effort. The lab-based animal digestion research described is 

indeed needed. If any actual animal feeding trials are executed, what value can be obtained given the 

limited research time available? 

The weakness of the program are probably due to the fact that it is only one year old and due to the 

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perceived need to cover all bases in the proposal. Down-selects should occur as soon as possible to help 

focus the program and reduce management burden. The down-selection process should be based on 

realistic transparent projections related to the points identified by the researchers and listed in the first 

paragraph above. 


This project is too large and the presentation too short for an adequate review. Overall communication 

appeared good, but it was not clear that the many individual projects would receive timely and objective 

evaluation by the managers and integration with the pilot- and full-scale production pipelines. With 50M 

in funding, this project should produce algal biofuel. Will it be able to make timely changes in R&D so as 

achieve that goal? 


As stated above, the program appears to be a relatively strong collection of project investigators who 

bring their technology to the program without an appropriate overall view of just exactly is the overal 

system of unit operations that will be applied, and how their technology should fit into the system. The PI 

needs to set the tone and gials, and not let individuals assume their technology is the anchor around which 

every other unit operation should bend. 


The project is too large and diverse to review based on a 1 hour presentation. All reviewer comments 

should be viewed in that light. We don't have a clear picture of the overall effort and the state of progress. 

The project is well-structured around specific quantitative goals and decision points. 

It would be valuable to reveiwers and to project managers to have currently achieved results compared to 

the stated project goal and to the pre-project state of technology for each activity. 

Within the constraints of the funding structure and the interests of the participating partners, increase 

focus on the biggest technical hurdles,--strain, cultivation, harvest, extraction and techno-economics. The 

program will not fail due to lack of technology to convert lipids to fuels. Some of the co-product studies 

appear to be premature because strain and process will dramatically affect co-product characteristics. Coproduct 

work should be limited to early feasibility demonstration. 

Algea screening processes need to be validated against pilot results to determine if the screening 

processes and parameters select superior strains for end use conditions. 

Having the project dispersed among many institutions does not appear to be a weakness. 


Yes, the consortium has already held several regular meetings during the past year, and will continue to 

do this in the future. We agree completely with the reviewer and stated during the presentation that the 

organism, Botryococcus braunii, is currently being sequenced by JGI. We did not try to infer any prior 

involvement in this process by NAABB until our organization was formed. The community of scientists 

that provided the materials for the JGI sequencing program included a team of investigators from Univ. of 

Arizona, Texas A&M, LANL, Northern Arizona Univ., and the Univ. of Kentucky. Except for the last 

two institutions, the others are part of the NAABB Botryococcus program, and these investigators are 

currently working with JGI to finish the genomic sequencing of one of the strains, and annotating and 

mining the sequence information that has already been provided by JGI. This is certainly something we 

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always have in mind in the Algal Biology and Algal Cultivation teams. As we identify promising genes in 

Algal Biology we will put them into model or production strains to test them against the baseline (parent 

strain). Our project on developing a pond-simulating photobioreactor that can be widely deployed across 

the NAABB will allow us to acquire standardized (comparable) rate and lipid productivity data from 

across the consortium that can be incorporated into our economic models. Within our 15 and 18 month 

(June and September 2011) deliverables the NAABB program is set to generate the baseline cases along 

with a preliminary assessment of all of the quantitative parameters for our harvesting and conversion 

technologies in order to determine feasibility for technical success. We recognize the need for full testing 

of strains under outdoor cultivation conditions. Our strain selection approach is multi-phased with a highthroughput 

screen that selects strains in the lab for growth and lipid productivity. Those that make it 

through this initial test then are taken to outdoor pond environments for further evaluation of their 

characteristics and robustness. Finally, we have to date isolated over 800 samples of algae, and screened 

over 400 in the lab for productivity. About 60 of these show excellent promise, and several of the strains 

are being tested in our small outdoor raceways. Stresses do need to be added to the second tier screening. 

We consider the efforts to be fairly efficient in using a combination of plating and FACS isolation of 

single algae. Selecting for ability to grow on solid and liquid media, exhibit reasonably fast growth rates 

in liquid culture and accumulate lipids are our first tier selections to quickly narrow down the field of 

candidates. The Solix Biosystems partnership is an important one for the consortium as they provide an 

excellent working process for commercial photobioreactors. This process will help us evaluate their 

feasibility in a number of applications, including the ability of PBRs to contain organisms along with 

issues of predatory and bacterial infections. With the recent placement of a Solix photobioreactor at 

NMSU, we will also be able to address a number of other issues associated with the best utilization of 

PBR’s in algal cultivation. Our objective for this peer review was not to describe any technology in detail 

but to provide an overview that would fit within the allotted time period. The wastewater and produced 

water efforts within NAABB have as major objectives understanding the nutrient utilization and recycle 

of such waters, but most importantly the contamination issues and clean-up processes that would be 

required to make this important source of water usable in algal cultivation. The requirement for large 

amounts of biomass is limiting NAABB’s ability to carry out experiments on large animals. The timing is 

now getting critical and major decisions will be made within the next couple of months as to what animal 

studies move forward based on availability of biomass. 

Project: 9.5.1.4 Cellana 

Title: Large-Scale Production of Fuels and Feed from Marine Microalgae 

Presenter: Jeff Obbard 



Approach 4.14 0.83 7 

Progress 4.29 1.39 7 






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What is the program? It was not clearly stated. One can look at the diagrams and still not follow exactly 

what the program is. There are too many disparate programs listed, but which is it that will be pursued? 

Alternatively, there could be more effective delivery by consolidation. 


Project has just started and has good targets/ goal schedule. Most data presented were based on a single 

diatom; screening of new strains (500) be fruitful--- but what are they? What are their tolerances? How 

much lipid do they produce? These aspects might be considered in future, given that the project is just 

starting. 

The productivity target may be unrealistic; productivity values that were presented were extrapolated to a 

large scale from a small experiment without any of the negative factors of large scale considered or 

explicitly identified (e.g., competitors, pathogens). Similarly, amino acid contents of several "novel" 

strains were presented by number alone (not algal group) and look as if they supply reasonable required 

amino acids but without control data from a terrestrial protein source or fish etc. were hard to evaluate. 

The project is promising due to integration at the Kona facility of all aspects (large scale growth to 

facilities for processing and biofuel separation). 


The focal point of Cellana is to develop new strains of microalgae with high productivity, demonstrate 

sustainable production of biomass and bio-product, efficient harvesting, all based on pilot-scale, and to 

further demonstrate end-to-end process integration. However, there are concerns regarding the proposed 

approaches by Cellana: 1- Random mutagenesis to generate and isolate strains with enhanced lipid 

production would not work. It is the wrong biological approach, the reason being that mutagenesis tends 

to eliminate single enzymatic steps or processes but it is not designed to induce enhanced generation of a 

specific biosynthetic pathway and accumulation of product. Moreover, chemical, UV or similar 

mutagenesis would introduce, with certainty, multiple (dozens or hundreds) of mutations that invariably 

lower fitness of the organism, resulting in lower overall growth and productivity. 2- Reference was made 

to algal "husbandry" by which to improve microalgal productivity. Aside from the bad choice of term, 

husbandry entails genetic crosses, something that is not known to occur in microalgae. Cellana ought to 

seek help with their proposed approaches to avoid going down the wrong path(s) in their otherwise 

valuable effort. 




Among a number of objectives, the most important is to be able to test and validate different culture and 

harvesting operations at production scale. Many of the approaches (such as strain selection, optimizing 

culture conditions, and harvesting/dewatering) seemed very conventional and unlikely to yield significant 

improvements in yield. 


Although the basic concept is reasonable - intensive innoculum development followed by deployment 

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into open ponds operated on a short interval - the PI did not demonstate much data supporting sound 

research and deployment, other than a multitude of screening work developed. Little was said about the 

large scale deployment and no project management plan was identified and the PI (Huntley) did not 

present with the facility manager presenting instead. Milestones were absent and not well defiend. 


The operation and optimization of the algae growth system appears to be technically sound, but the 

approach, structure, and activities of the project appear to lack critical, objective review in the context of 

large scale biofuel commercialization. The current effort appears to be directed at small-scale, high-value 

algae products, and not at large scale biofuel production. 






Progress appears adequate to the extent of the program thus far. 


The project is just beginning and only Cellana has begun initial work. 


More than 500 strains have been isolated and screened. Some have been subjected to pilot-scale growth 

and production, as well as techno-economic and sustainability analysis based on (presumably) a prior 

3+years of Consortium result. 




The project was funded in Sept 2010 and results were very preliminary. Some of the results seemed to be 

extrapolated from small scale experiments and assumptions were not always clear. 


Other than strain isolation and screening, little progress was identified other than statements of 

achievements, some of which did not stand up under questioning. Milestones related to production of fuel 

were made (gallons per acre per year) that were misrepresented. The PI should include all steps in the 

growth process, including the area and time devoted to the production of the innoculum should be 

included in the calculation of biomass productivity. One cannot only use the time that the innoculums 

spend in the final grow out ponds. Finally, water use needs to be presented in terms of the amount of 

product used. One should not say a goal has been met by reducing water use to 10,000 m 3 . This term has 

to be relative to some measureble resource utilzied. 10,000 m 3 per WHAT? 


The strength of the progress is the operation of an outdoor system producing 1 metric ton/month dry algae 

biomass, but this needs to be viewed in the context of 1000 MT/day needed for an at-scale biofuel plant. 

Data presented on productivity and economics lacked sufficient detail to be meaningful in the evaluation 

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of progress. Hidden assumptions in productivity numbers and project economics further obfuscate the 

state of progress. 







It is hard to guess the relevance. If the deliverables were better defined, they may be compared with the 

OBP program goals. 


The projects goals align with the Biomass program. 


The Cellana project addresses operational aspects of microalgal cultivation, cost of production & process 

sustainability 




The project is highly relevant to program goals. 


It's relevant to look at marine strains and Hawaii is an appropriate place. In terms of applicaitons, the use 

of defatted biomass as an animal feed is overlooking the reality that lipids are the desired nutrient of fish 

farmer. So the real value for feeds is in the lipids, specifically those with neutaceutical value. Was this 

considered? 


Operating and optimizing a real world cultivation system is essential for the potential development of 

algae biofuels. 

Specific project work on economics and co-products are not relevant to commercial scale biofuel 

production. 








assessments) 


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end-use) 

iv. Environmental sustainability (example: water use, GMOs, energy consumption). 



Goals are listed rather than CSFs. 


Some factors are well-considered at this early stage (e.g., ability to return seawater without nutrient 

enrichment to sea due to algal N depletion to cause lipids to rise). Others need more attention. For 

example, more attention is needed on the effect of different strains and conficting goals of lipid extraction 

on byproducts (e.g., aquaculture feeds). Algal strains lacking high carotenoid levels are needed for 

production of fish pellets for cod to prevent them being pink, not white....a market problem. A major 

current deficiency in aquaculture feeds for marine fish is to supply omega 3 fatty acids---this is leading to 

overfishing of small fish and krill to supply protein/omega 3 fatty acids to salmon etc farms. It would be 

helpful for this young project to address how problems such as this can be addressed to produce valuable 

byproducts (e.g. aquaculture feeds) while producing biofuels in a sustainable manner. 


As with the other projects, these revolve around microalgal productivity, efficiency and cost of 

harvesting, downstream carbon intensity, and aquafeed product value. Cellana should talk to RAE, as 

they have developed an apparently efficient way of harvesting algal biomass. 




Critical success factors were described but it was unclear whether the project will overcome these factors 

using traditional approaches. 


The presentation of the data was "couched" in terms that were misleading. 

The group has no extraction technology put in place and no obvious mechanism to deal with the salt that 

will be most prevalent. 


Critical success factors are identified, but they are defined in the context of producing a high value algae 

product. A limited amount of technology developed is likely to be applicable to large scale biofuel 

production. 





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Publications are in hand already. The project appears to be self-contained. What comparisons with other 

oil production rates are expressed? No simple comparisons can be found. 


This is a new project and does not yet have tech transfer. 


Cellana should talk to RAE, as they have developed an apparently efficient way of harvesting algal 

biomass. 




The potential for technology transfer is good. 


The project seemed fine in this area. 


It would be very valuable to share the results of screening 500 strains of algae, and how the screening 

results compare to real world productivity. Because of the focus and reliance on high value co-products, 

the design report will likely have limited value for advancing large scale biofuel technology. 






My overall impression is that there could be more value to this project that could be achieved. There are 

questions about the ways the cycle time is being calculated, and some comments about how yields are 

expressed that also detract from perceptions. 


This is a project with great potential; its success depends upon transparency, accurate comparisons etc. 

These factors are required to recognize problems and correct them. 


Slide 12 on productivity claims "Sustained Productivity" of 200 MT dw per ha per year. How do they 

propose to achieve this, when best-case scenario on sustained productivity of microalgae is 20-30 g per 

square m per d (~100 MT dw per ha per y)? There are concerns regarding the proposed approaches by 

Cellana: 1- Random mutagenesis to generate and isolate strains with enhanced lipid production would not 

work. It is the wrong biological approach, the reason being that mutagenesis tends to eliminate single 

enzymatic steps or processes but it is not designed to induce enhanced generation of a specific 

biosynthetic pathway and accumulation of product. Moreover, chemical, UV or similar mutagenesis 

Page 24 of 223




would introduce, with certainty, multiple (dozens or hundreds) of mutations that invariably lower fitness 

of the organism, resulting in lower overall growth and productivity. 2- Reference was made to algal 

"husbandry" by which to improve microalgal productivity. Aside from the bad choice of term, husbandry 

entails genetic crosses, something that is not known to occur by, let alone controlled in microalgae. 

Cellana ought to seek help with their proposed approaches to avoid going down the wrong path(s) in their 

otherwise valuable effort. What is 4 in-pond harvesting processes?? 3 out-of-pond harvesting processes?? 

These need to be explained. 



The cultivation ponds and ability to conduct aquaculture feed co-product studies are impressive. A major 

problem was that the presentation gave the impression that Cellana was achieving 100-150 MT/ha-yr of 

algae production in the ~1000-m2 ponds depicted in the aerial photo. Upon inquiry, the presenter clarified 

that that productivity was achieved over the short term in 200-L tanks. In addition, total dry matter was 

presented, not the relevant characteristic, ash free dry weight. This came across as an effort to mislead the 

reviewer panel and DOE. In addition, the target of ~200 MT/ha-yr was presented but without any serious 

discussion of how the capability to achieve that extraordinary productivity would be developed. 

"Hydrodynamically-induced flashing light" was listed as one of the methods to improve productivity, but 

this classical effect is operative only at the milli-second scale, and the energy consumption to achieve that 

scale through mixing would be too large for a good energy ROI. 

In-pond sedimentation and "sweeping" was touted as a major advance, but the equipment needs and costs 

to make use of this process at the 1000-acre scale need to be projected to determine if it is feasible. Other 

issues not addressed include: 

Wet extraction studies were conducted, but what was the solvent loss rate? What is the ratio of PBR 

ground coverage to pond area, and how does that relate to overall areal productivity and costs? 

The presentation stated 3 years of consortium operation, $90 million invested, 20% completion on the 

DOE project, but the main results seem to be construction of the facility (good). Very few data were 

presented for a project touting this level of maturity. How was the figure of $90 million invested arrived 

at? 


The presentation described a very conventional algal production process. The primary value of funding 

this facility is its potential to test different processes at a large scale. Although it is very early in the 

project, I am concerned whether the 9M in funding will yield results that will be useful to the biomass 

program. 


The progress presented seems very small compared to the amount of money invested (over 90 million). 


This project may have great value in developing algae of aquaculture feed, but it is not on a path to 

commercialize large scale biofuel production. 

The value of the whole algae production optimized to aquaculture feed needs to be rigorously compared 

to the value of biofuel plus lipid extracted algae meal. It appears very likley that the economics are much 

better for optimized whole algae aquaculture feed than they are for biofuel production with coproducts. 

Page 25 of 223




Project: 9.5.1.5 Arizona State University 

Title: Sustainable Algal Biofuels Consortium 

Presenter: John McGowen 



Approach 7.43 1.18 7 

Progress 6.71 1.28 7 










The Approaches are well developed and firmly tied to expected outcomes and deliverables. 


This project concentrates on feedstock development including developing a matrix of algal biomass 

produced under different growth/processing conditions; screening for lipases and application of 

commercial lipases; develop biochemical strategies for conversion of lipids to FAME etc. (the latter just 

starting). 

Excellent work to date, especially in terms of careful QC evaluation between labs of different partners 

and within labs, of different technicians. This is a model for all DoE funded projects, and helps to assure 

success of the project. 


SABC will examine multiple biomass samples to explore the possibility of extracting lipid and 

carbohydrate-based biofuels. Theirs is essentially a compositional analysis methods development, as well 

as development of methods for evaluation of productivity in growing algal cultures. Pretreatment steps 

will be employed that may eliminate need for drying biomass and offer lower cost and lower energy route 

to lipid extraction and conversion. Enzyme discovery will supplement commercial enzyme evaluation to 

facilitate conversion with heretofore unexplored biomass. SABC ought to talk with RAE, as they have 

developed method for the effective harvesting of microalgal biomass. 


See Overall Impressions text. 


Project approach and direction are sound and were presented well. 


This is a precisely focussed project, with obvious data and material controls put in place. Very smart to 

Page 26 of 223




look at whole cell conversion (i.e. use algal biomass similar to other biomass) coupled with the immediate 

testing against ASTM standards. The deconstruction of biomass is usefull as well (for eventual enzymatic 

hydrolysis to simple sugars) and couples this work with the tremendous amount of work already done on 

biomass (in this area). It is rather apparrant the system rules in this program. 


Excellent approach to working toward standardization of algae analytical techniques. 

Specific quantitative goals for the conversion processes should be defined and expressed. These goals 

should be based on a clear definition of quantitative feasibility parameters. Criteria for milestone decision 

should be defined based on these quantitative commercial feasibility goals. 


Thank you for the supportive comments and constructive feedback. 

We agree with the reviewer that establishing both goals and metrics for the conversion process will add 

value to the project. To compare conversion conditions that will allow down-selection to more effective 

conditions we’ll focus on component yield: 1. Mass of sugars released from pretreatment and enzymatic 

hydrolysis of algae. Treatments releasing ≥50% of the initial algal carbohydrate composition will be 

selected for further study. 2. Mass of lipid extracted from algal biomass either before or after pretreatment 

and enzymatic hydrolysis, based on a statistically relevant difference (≥20%) between treatments at 95% 

confidence level. These two metrics will allow for selection of conversion processes that will lead to 

deconstruction of algae providing both high lipid and carbohydrate removal across different growth 

regimes. While these criteria may identify effective pretreatment and enzymatic hydrolysis processes, it 

may not possible to link these targets to quantitative, commercial, feasible goals based on the lack of an 

industrial standard process. 

Additional criteria for measuring both goals and progress in the conversion task related to both enzyme 

discovery and enzyme efficacy in lipid/cell wall deconstruction are more difficult to quantify. 

Examples for a minimum benchmark for enzyme discovery: 1. Selection of 10 thermophilic fungal strains 

isolated from targeted screens, demonstrating enzymatic activity using processed algae (control, 

extracted, pretreated) for genomic sequencing. And for enzyme efficacy: 1. Demonstrate activity 

(protease amd or lipase) based on a statistically relevant difference (≥20%) compared to control 

conditions, at a 95% confidence interval. 






The project has not been in operation long enough to generate progress. 


Good progress and milestones are carefully spelled out. Project seems well managed and coordinated. 

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Too early to tell. 




Progress thus far was mainly on QA/QC issues between labs. Project plans were sound. 


Fine for time funded. 


Good progress is evident in standardized characterization of algae biomass. 

It appears that the basic criteria for measuring progress in pre-treatment and biomass conversion are not in 

place. 

Growing biomass appears to be a bottleneck. Partnership with facility that can supply kg quantities might 

be valuable. 


The primary criteria for measuring progress in pre-treatment and biomass conversion for this project will 

be based on the mass of both carbohydrate and lipid removed/extracted from algae grown under different 

growth regimes (See above section in Approach). Coupling the advanced characterization method 

development and leveraging the experience within the team in quantifying cellulosic biochemical 

conversion, along with the detailed biochemical fingerprinting (QA/QC) enables the SABC to develop 

and then apply critical tools to allow for quantification of the biochemical conversion process in algal 

biomass. Defining the criteria and developing the tools to help quantify the biochemical conversion 

processes are a critical early objective for the project. 

Biomass supply in the kg range will come from the lead institution ASU. We have devoted significant 

time and resources into the biomass supply pipeline to ensure adequate supply. 

We are confident in our ability to meet the needs of the project as defined. However, we are always 

actively exploring where we can expand the potential sourcing of biomass especially as we look to year 

two activities and or if we progress faster than expected and have the desire/need to take things to a larger 

scale or look at additional production strains - especially from [pre]-commercial sources (especially from 

any of the other DOE funded consortia). 







The project has the potential to be extremely relevant. Investigators have considered this aim in the 

program definition. 

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Bench-top scale is the major limitation, but project personnel indicated their hope to provide their data on 

best feedstock species/growth conditions and processing for biofuels to other DoE projects that could 

implement at larger (production-scale) facilities. 


The work provides a basis for integrated algal bio-refinery operations while offering a broad biofuel 

portfolio. 




The project goals are highly relevant to the Biomass Program plan. 


Most relevant given that focus on whole cell use for fabrication of a range of high value fuels (not just 

extraction of bio-oil for biodiesel). The deconstruction of biomass fits much of the work being done in the 

biomass program. 


High relevance for algae characterization work. Rational design of process is contingent on having well 

characterized feedstock. 

High relevance for biochemical pre-treatment. This addresses a major hurdle in unlocking economical 

value of algae slurry. 

Work on converting FAME, FFA, and sugars to fuels is not relevant to overcoming hurdles to algal 

biofuel commercialization. Algal biofuels will not fail because of fuel conversion process viability. 


While we acknowledge the general sentiment questioning the relevance of the work converting FAME, 

FFA, and sugars into fuels as not relevant to overcoming the hurdles to algal biofuel commercialization, 

we feel some of the nuance of what it really takes to take a new feedstock and make fuel (new or old) may 

be overlooked in that sentiment. Neither the FAME, FFA, nor sugars produced from algae or any other 

biomass source will be 100 percent pure or reagent grade. The small portion of the project devoted to 

making and characterizing finished fuels will evaluate the potential for these impurities to 1) negatively 

affect conversion process chemistry or operability and 2) negatively affect fuel performance. 

Additionally, for some fuels such as biodiesel or renewable diesel, both made from lipids, the feedstock 

chemical structure can have a significant impact on the final fuel properties, potentially in a negative way 

that would create additional barriers to commercial deployment. While not the main focus of the project, 

knowing if these issues exist early on will be critical for more rapid commercialization of algal biofuels. 




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assessments) 




end-use) 




The CSFs are enumerated more as the deliverables of the work. That is not too objectionable, given the 

thoughtful analysis leading to the program. 


Continued attention to detail and QC is important. Interaction with an appropriate DoE-funded project(s) 

that have production scale facilities towards end of project would be ideal. Perhaps program managers can 

help here. 


Project aims to develop a suite of rapid, accurate, and reproducible analytical methods for determination 

of algal biomass composition that can be applied to whole cell biomass as well as fractions generated by 

processing of the biomass. Effort further aims to apply optimization of biomass pretreatment, including 

enzymatic hydrolysis and conversion operations of the biomass with integration into a single, efficient 

process. These items are a tall order to be achieved in this short period of time. 





High marks here. In particular the use of ASTM standards from the get go. Correlating that to the profiles 

of the strains grown is appropriate. The program becomes even better when it considers natural wild 

mixed cultures instead of pure strains. 


Plans for earliest possible techno-economic modeling to produce quantitative goals for feasibility 

demonstration is essential. 






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This cannot be scored yet. For success, there must be coordination, and there will be significant use of the 

results. 


Good goals for this. 








Very well organized and appropriate, actually excellent balance between NREL, Sandia, and ASU. 


Excellent cooperation with external institutions during robin development of analytical procedures. 

Good sharing with many results public. 







There is significant potential with this project. It is extremely well designed. 


If project continues with current scientific rigor and pragmatic outlook, high success is possible. 


Interesting presentation. What is the rationale for the selection of Chlorella, Scenedesmus, and 

Nannochloropsis in this work? Definition of the species and strains is important for the proper assessment 

of the effort and its outcomes. Please identify the precise strains employed. Overlap with the work 

performed in other supported projects? 



The focus is biochemical conversion of algal biomass, but the presentation acknowledges that biomass 

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composition is a function of species and growth conditions. Of the limitless combinations to test, their 

focus on "production" strains is sensible, but still the growth conditions and any hypothesized 

mechanisms to control composition are not discussed. In addition, what are the backup plans in case 

biomass production falls short of the needs for the rest of the research? 

The lab QAQC and analytical methods development work will put the project on a good footing from the 

start. More data were presented from this young project than from more established projects. But what are 

the details of the plans to develop real-time high-throughput methods for compositional analysis? 

Projections should be prepared on the economics of using enzymes, ionic liquids, etc. to be used as a 

baseline to evaluate performance and long-term prospects of the processes being developed. An estimate 

should be made for each process and envisioned integrated processes. The initial projections will no 

doubt be unfavorable, but this will help in planning the course of process development. Economic 

performance projections should be re-evaluated periodically as the results come in. 

The commercial partners should be expected to allow complete review and inspection of the data they 

generate in order to maintain confidence in the results. 


Very nice presentation of a multi-institutional project. 


The best consortium presentations by far. 


The concepts of hydrolyzing and fermenting algae and lipid extracted algae are potentially enabling for 

algal biofuels. To determine how far the concepts are from commercialability quantitative goals for 

feasibility demonstration need to be defined. These should be based on preliminary techno-economic 

modeling and should include parameters such as yields, payloads, cycle times, enzyme costs, feedstock 

requirements. 


The rationale for the choice of strains for this work is based on 1) experience within the team, 2) overlap 

with existing programs, and 3) the fact that these strains are comparable to the best biofuel production 

strains in the literature (i.e. it is a combination of strain availability and experience with these strains in 

our indoor and outdoor production environment, growing them under different conditions, and at a scale 

that allows for high confidence in our ability to deliver the necessary biomass quality and quantity to the 

team). The strains are all maintained as part of the strain collection within the Laboratory for Algae 

Research and Biotechnology at Arizona State University and are all publically available. We will disclose 

the specific strains (and source) to the DOE and upon publication of results which will include the growth 

conditions utilized to achieve different biochemical compositions for a given strain. As discussed above 

in section 2, we are confident in our ability to supply the necessary biomass to meet the needs of the 

program as currently defined. 

As to the question around detail plans for real-time/high-throughput compositional analysis, based on 

encouraging preliminary results within the partner institutions, we will continue to develop, LC/MS 

analysis, Infrared (IR)-based spectroscopy methods and chemometric models for rapid compositional 

analysis with the goal to make these methods the default compositional analysis methods. Application of 

these methods may reduce compositional analysis time from days to less than an hour (e.g., NIR based 

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methods) and can also significantly reduce sample size (e.g., LC/MS). The results from these analyses can 

guide pretreatment and enzyme testing and development tasks and lead to a down-selection for strains 

best suited to fuel production (i.e. highest lipid and carbohydrate composition). These chemical 

compositional analysis studies will reveal to us what chemical additions to the pretreatment step (acids, 

bases, etc.) will be effective at recovering some of the sugars and assisting in the detailed process 

characterization around the different algal deconstruction, enzyme discovery and efficacy pathways we 

are exploring. Having an idea about the carbohydrate make-up of a cell is going to suggest potential 

enzyme preps that may be useful in the hydrolysis step and we feel this work will be equally applicable to 

whole algal biomass and to the algal biomass that remains after lipid extraction. 

The several constructive comments about techno-economic analysis throughout this review we 

acknowledge are spot on. We are focused, in a broad sense, on the ultimate goal of reducing costs to 

allow for economically viable and usable products, including biofuels, from algae. However, in the 

current funded scope we have acknowledged that we will have to leverage our technical results and make 

our data available to the many programs funded by the DOE that already are established in order to look 

at the technoeconomics of algae biofuel production - but may benefit from additional data to feed and 

exercise the models they are generating. The information and data we generate we will continue to 

actively seek to share within the community and ideally leverage, especially through the large-scale 

projects funded by the DOE such as the other algal consortia, the significant emphasis and defined scope 

of which is already established by the DOE for technoeconomics of biofuel production from algal 

biomass. 

Project: 9.5.1.6 CABComm 

Title: Proposed research activities for the Consortium for Algal Biofuels Commercialization 

Presenter: Paul Falkowski 



Approach 5.17 0.90 6 

Progress 5.33 0.94 6 










The approach seems reasonable for such a disparate, multi-pronged program 


Project has just started. Presentation did not outline goals and management strategies clearly. While many 

good people are involved, some concern was raised by the presentation, which stated that award monies 

would be evenly divided among PIs; some goals, especially the larger-scale ones that are so critical, 

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might require more funding and sub-groups that were not evident in presentation. 

PO4-3 is indeed important, but is only one piece of the puzzle. The presentation flashed a slide of other 

projects that are included in the contract, but they were not explained. 


In consultation with Commercial Partners, CAB-Comm will: Identify the most pressing research needs 

and assign a priority status: Identify the research group(s) with the capabilities and expertise to address 

each priority item: Identify milestones and deliverables for specific projects 


Not possible to adequately score this item because of inadequate detail in presentation. There were no 

detailed descriptions in the presentation about the role of each laboratory. In final slides, 5-6 general 

objectives were listed for each of the three goals. 


See Overall Impressions 


Milestones and deliverables were not defined. Targets of research are broad and vague. 







No actual progress expected because of the very recent actual award. At least plans are being formulated. 


Just started. 


Too early to tell, project just started (last week). 


No progress to date--funding just begun. 



Since this is a new project, the expected progress is to have defined objectives and a management plan. 

These were not presented. 

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The directions presented will be relevant if successfully addressed and completed. 


Only partially explained in presentation. 


The three research focus areas of CAB-Comm fall within critical stages of the overall algal biofuels value 

chain. 


Not possible to adequately score this item because of inadequate detail in presentation. 




Crop protection, nutrient utilization and recycling, and genetic tool development are essential for algal 

biofuel commercialization. The project has not defined expected outputs. 









assessments) 




end-use) 


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Some listing of legitimate CSFs was presented. 


A strong management structure that links PI's goals and ensures a focus on end goals is critical (and was 

not apparent from presentation). 


Critical success factors to technical and commercial viability: Integration of research projects with 

commercial partners. Achieve milestones and deliverable on time. Continuous review of projects to retain 

commercial relevance. Potential Challenges to overcome and to achieve project results: With 21 academic 

participants, communication between labs and commercial partners will be essential. With only two 

industrial partners, commercial research focus will be much easier to mamage. Demonstrate how project 

will advance the commercial viability of biomass: Sapphire projects are directly tied to viability of first 

integrated algal biorefinery - due to start operations in 2012. Life Technology projects will enter market 

as research tools in 2011. 


Unfortunately, there were no detailed descriptions about how achievement of milestones will change or 

not change goals 

or collaborations between laboratories. 




No evidence of indentification of critical factors as defined above was presented. 








Coordination will be one of the big challenges for this program. 


Just starting. 


CAB-Comm works closely with Sapphire Energy in the area of Tech transfer 

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Potential for technology transfer is good given that this is a multi-institutional collaboration. 




Within the consortium there is a broad group of academic and industrial partners. No plan for outside 

communications and co-operation was given. 







This project has potential. The investigator was not happy with the reduced scope, but it is hoped this 

becomes a non-issue in favor of real progress. 


Project needs integrated milestones and goals to succeed. PIs can be expected to do good basic science. 


Interesting and well handled presentation. Telling it like it is without exaggeration and false promises. 

Impressive group of investigators in CAB-Comm, increases the credibility and likelihood of success of 

this effort. The planning is good, critical progress and direction control items are in place and, therefore, 

there is a strong likelihood of useful results emanating from this effort. 


Since this project just began, it was inappropriate to evaluate progress. However, the presentation lacked 

descriptions about the role of each laboratory and how achievement of milestones will change or not 

change goals or collaborations between laboratories. 


As this was just funded it is very difficult to review. It is clear that the presenter has a good perspective on 

the field (he nailed the P issue on the head) but the "world is about to collapse" terminology could be 

avoided as a motivational tool. Also, funding each PI 100,000K equally is likely to keep this a program 

that only produces individual work, by each PI, and could undermine any chance of building up a true 

consortium that develops a process. 


Although this is a new project, the presentation of scope, goals, and approach did not meet minimum 

expectations. 

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Project: 7.2.1.7 University of Nebraska Lincoln 

Title: Research for Developing Renewable Biofuels from Algae 

Presenter: George Oyler 



Approach 3.29 1.28 7 

Progress 3.86 1.46 7 










Some of the basic studies were desirable approaches. Some of the extensions, such as the Photobioreactor 

system or the nanobody approaches are far less justifiable. 


Focus is on developing photobioreactor with new/improved processes for concentration of algae/lipid 

product. The project is developing a lipidome and transcriptome for relevant conditions (N deprivation, 

acetate free media etc.). Evaluation of algal viruses is being made for a processing step. Bioreactors of 

this type may not be efficient for scale-up and costing of processes and procedures needs more analysis. 


[1] Project aims to increase fundamental knowledge of lipid biosynthetic pathways, lipid transport, and 

storage and secretion mechanisms in two important eukaryotic algal genera, Chlamydomonas and 

Chlorella; [2] Develop new and improved technologies for manipulating genomes and gene expression in 

eukaryotic green algae; and [3] Establish a state-of-the-art Photobioreactor Research Facility to develop 

low cost, efficient algal cells harvesting strategies and to improve current technologies by developing 

novel, next generation photobioreactors for algal biofuel and specialty chemicals production. 




Goals are focused on lipid production, photobioreactor development, and harvesting. Novel aspects of the 

project include use of viral enzymes for algal lysis and nanobodies for algal harvesting. The 

photobioreactor research appears to ignore the results from decades of work by other groups. 

Unfortunately, the nanobody approach appears to be wildly unrealistic for real world conditions. 

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In addition to developing state of the art photobioreactors, the PI's suggest to tether cells to cellulose 

membranes. As a laboratory excercise on a nano-scale, this is most interesting, but unrealistic commercial 

scale production. In addition to the costs of cooling and materials, biofilms will coat the internal walls of 

any photobioreactor. Such films will restrict photon transfer and undermine the need to tether the cells. A 

simpler approach would be to innoculate with metaoblically manipulated cells that secrete oil and allow 

them to coat the inner walls. Still, cooling costs will likely challenge this approach. There is also the issue 

of CO2 delivery to tethered cells within a commercial scale photobioreactor system. 


Well-defined criteria for success and milestone decision points were not presented. It did not appear that 

the scope was defiend by a preliminary evaluation of commercial viability. 


xx 






On the basis of the results in the presentation, I believe more progress on the key thrusts could be made if 

less attention was given to nanobodies, etc. 


This project is 50% complete. Membrane development and antibody stages of processing development 

appear expensive and their effectiveness for scale-up, especially with algae growing under commercial 

conditions needs to be demonstrated more rigorously. 


Not a whole lot of progress yet. However, the effort aims to address: 1. High resolution lipidomics 

analysis capacity developed, 2. Transcriptomic analysis of N deprivation underway, 3. Development of 

novel molecular tools for genetic manipulation of algae, 4. Advancement of molecular algae viral 

pathogenesis, 5. Development of nanobody tools for algae growth, harvesting and research. 




The project appears to have met some of its objectives (development of molecular tools, etc). 


A fair amount of data was presented. 


Data compared against a well defiend criteria to measure progress were not presented. 

Page 39 of 223











There are some good outcomes that have relevance. There are others that do not. 


PIs should refocus on core goals/milestones and do costing/scale-up calculations to justify that this is a 

sustainable biofuels' production process. In the absence of this, the types of processes and materials being 

developed and the bioreactor system itself raises conerns about the amount of energy input required to the 

biofuel's derived energy that will be available. 


- Development of an understanding of TAG oil formation in lipid bodies after N deprivation of 

microalgae, including de novo synthesis vs. redistribution of membrane lipids. - Development of novel 

nanobody based membrane linkage and cell harvesting systems. 




Unfortunately, the project's approach and results do not seem relevant to meeting platform goals and 

objectives. 


One cannot be relevant until the work is appropriately placed in the reports by Benneman and co-authors. 


The engineered nano antibody work does not appeear to have commerical potential at scale. 

The reasoning for pursuing PBRs was vague and non-technical and did not show a pathway to overcome 

cost issues for PBRs that have been shown in techno-economic modeling. 








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assessments) 




end-use) 




These were not really addressed in the presentation. The project could use a review of these even at this 

stage of completion as part of a consideration of what the valuable deliverables should be. 


Demonstration of energy efficiency and sustainability of processing technologies under development is 

very important. 


Transition of basic molecular research to algae biofuels and bioproducts is a challenge that is being 

addressed by team approach involving multiple disciplines 




The presenter was not successful in convincing me that this project would advance the state of biology or 

technology relative to platform goals. 


Most hurdles to photobioreactors were not well addressed in the data. The PI should revisit reports by 

Benneman and co-authors. 


Project viability criteria based on commercial requirements were not presented, and no plan to overcome 

known barriers to commercialization of PBRs was given. 








There have been a number of publications. There had to be internal coordination during the project. It is 

not clear whether the plans and results have been extensively shared within the OBP program 

participants. 

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Required additional work and documentation. 


UNL directed spin off underway for algae biotechnology and biofuels (NU Green Technologies). Joint 

ventures with SR, CGC, NPPD resulting from this funding. Provisional patents filed for aspects of this 

funded research 




No technology transfer activities were detailed. 


No comment. 


Prolific publication ensures tech transfer. 







The investigators should close out by considering the detailed outcomes in hand and how they can 

advance the overall program. Starting new lines of inquiry into photobioreactors and other exploratory 

programs are not warranted. 


Project appears to be following potentially interesting basic science directions but without as much focus 

on core milestones (sustainability, practicability, scaling) as is important. This reviewer has particular 

concern over the use and subsequent ability to eliminate algal viruses from the process; PIs should give 

this more attention in terms of scale/pragmatic concerns. 


A multifaceted approach that is based on the expertise of current faculty members at the ULN, ranging 

from microalgae, viruses, molecular biology and photobioreactors. Strengths: the range of topic covered 

by the team. Challenges: integrating the currently existing expertise for the benefit of the Program. 



Not enough information was provided for this reviewer to evaluate the results of -omics, virus, and 

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genetics aspects of the project. 

The nanobody research is meant to help in harvesting of algae, biofilm growth of algae, and provide a 

research tool. Algae can be flocculated and can grow in biofilms without nanobodies, so the importance is 

not clear. Nothing was said about the potential research applications of nanobodies. 

The slides do not provide much information about the photobioreactor research. However, PBRs were a 

larger component of the oral presentation. The speaker was not able to effectively defend the use of PBRs 

for biofuel production. The well-know problems include excessive total costs, the likelihood of negative 

energy ROI, and difficulty in controlling heating and gas exchange. When asked about the method of 

cooling, the speaker said that evaporation of the water will cool the system but that water loses would be 

minimal due to the double-layered design of the PBR. Of course, if the water re-condenses within the 

system, heat is not removed. The speaker did not appear to have the basic knowledge needed to design 

PBRs. 


Although I agree that basic research may lead to improvements in strains and downstream processing, 

there was no aspect of this research that seemed likely to yield such improvements. 


Tethered microalgal cells and photobioreactors for commercial scale production of microalgae is severly 

challenged. 


From a process engineering perspective the project has failed to define a method for evaluating the 

commercial viability of concepts and has not defined criteria for success and decision points based on 

commerical scale processing requirements. Because proposed technologies of PBRs and nano-antibody 

structures have well defined techno economic hurdles, a clear pathway to overcome those hurdles is 

needed before meaningful research commences. 


The team at UNL, the PI, and the presenter appreciate the efforts and contributions of the review group in 

facilitating the achievement of the goals for the DOE algae biomass platform. While recognizing the 

contributions of the review team and the demands placed by the need to review a large number of projects 

in a short period of time, it is clear that the format of a 15-minute PowerPoint presentation followed by 5 

minutes of questions is apparently inadequate for the reviewers to grasp the scope and vision of the UNL 

programs in algae biofuels and biotechnology. First the UNL team recognizes the importance of assessing 

the short-term needs of open raceway production of algae biomass as an initial step toward understanding 

the economical growth of algae at scale and the challenges faced with scale-up. These steps towards 

growing algae with 30 to 60 year old technology but at a much larger scale are indeed meritorious in 

actually achieving experience of algae production at 10 to 1000 hectare scales. In this context the UNL 

team applauds the landmark works by Dr John Benneman in the Aquatic Species Program that 

summarized the technology of 1980 to 1996 and predicted the production of algae biofuels at $60 per 

barrel. We also are well aware of the recent highly detailed and very sound techno-economic analysis by 

Dr Benneman and one of the reviewers that suggest it is impossible to produce algae at less than 

$300/barrel as of the review in 2010 with current technologies of open raceway growth. We also are 

aware of the recent techno-economic reviews that suggest economic biofuels production with algae is 

nearly impossible with raceway design, hexane extraction (a well known neurotoxin), and 

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transesterification. The team at UNL concurs with these well founded reviews and recognizes the critical 

importance of basic research on lipid production in algae, the development of molecular tools for 

engineering algae, and the importance of low cost innovative system of growth systems utilizing highly 

innovative and patent pending tools such as nanobodies in order to allow algae biofuels to be a reality. 

We indeed accept that the substantial innovative approaches marrying world class basic algae biology 

research and transformative application of novel tools to growth and harvesting (nanobodies) that the 

UNL team is pursuing are critical to truly economical biofuels production from algae. As stated in the 

review oral presentation, the presenter is clearly aware of the limitations of open raceway systems with 

aerial productivity limited to 25 gm/m2/day which has remained constant over 40 years. The presenter 

indeed is aware of the severe constraints of photobioreactor designs (PBRs) as currently configured as so 

acridly depicted by the reviewers. The presenter is aware of the heat removal need in PBRs and correctly 

presented a number of means of such heat removal. The UNL team is in fact developing innovative low 

cost growth systems to address the limitations of current PBRs including the use of nanobodies. The 

transformative value of these breakthrough technologies will be apparent as the result of this well placed 

DOE sponsored research. Unfortunately these tangible advances were evaluated on the basis of limited 

information (a 22 slide PowerPoint) and through the lens current and limited raceway technology. The 

UNL team does hold the Benneman point of view in high regard, that maintains with current technology 

algae biofuels are economically unattainable. The UNL team also strongly believes in the ability of the 

combination of basic research and transformative technologies such as nanobodies to overcome these 

obstacles. The UNL team is grateful for the opportunity afforded by the DOE to play a role in this 

transformation of algae biofuels to become a major element in the DOE biofuels portfolio and regrets the 

focus of the review team on current technologies with all their limitations. 

Project: 9.6.5.3 NREL 

Title: Algal Biofuel Pathway Baseline Costs 

Presenter: Andy Aden 



Approach 8.14 0.64 7 

Progress 8.00 0.53 7 










This is a project that is producing useful results right from the start. The needs and deliverables were 

thought out first, and the approaches are devised to achieve them. 


Models are being developed well for evaluation of capability of sustainable production with most key 

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factors incorporated into bioreactor, open pond, and heterotrophic production models. Application of such 

models to other DoE projects in this area offers potential for greater transparency and sensitivity analyses 

in other projects. 


The goal of this task is to develop baseline technoeconomic analysis and user models for algal biofuels 




The presentation described the development of excel based program with process components identified. 

Very logical approach and well presented. 



With baseline cost modeling complete, additional milestones for incorporation of processing concepts and 

projections of costs should be defined. 


We agree that these models will be useful to a number of stakeholders within the research and 

development community. We look forward to working with the other projects so that we can be mutually 

beneficial to one another. We agree that additional milestones for incorporation of concepts and cost 

projections should be defined; this is being done in conjunction with DOE and others. 






For the short time in operation, results are already forthcoming. 


Relevant literature being used well to create models/factors included in models. 


Models currently calculate high costs for algal biofuels. Submitted publication to peer reviewed journal 

for autotrophic pathways (Applied Energy) 




Tasks appear to be completed. 


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Baseline modeling is already a valuable tool for guiding research. 


Thank you. 







The relevance is right on target. 


Models that can be used as part of reporting requirements to DoE are definitely needed and this project 

may supply them. 


Primary cost drivers are lipid content and yield. 




The project is highly relevant and provides very good benchmark values. 



Baseline cost analysis and extension to conceptual process cost projection is essential to rationalizing 

research and development resources. 


Thank you very much 







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assessments) 




end-use) 




The CSFs and Challenges are discussed in the presentation. 


PIs should continue to monitor literature to adjust models. Add additional factors (e.g., effects of climate 

change on precipitation/water availability to projects, sealevel rise to coastal land availability---consult 

USGS personnel over this point). 


Models will be useful in assisting DOE with target development and research interaction. Success 

Factors: Maintaining close interaction with researchers is critical, as is transparent communication of all 

assumptions and results to ensure proper use of data. 





Modeling still too removed from actual applicaiton. Need to integrate modeling with the consortium 

projects. 


The critical factor of access to pilot data for various process concepts is identified. 


We will continue to monitor literature to keep the models up to date and relevant. One of the first items 

we are working on next is syncing up with other resource assessment activities, such as the PNNL models 

for resource evaluation. We have already begun to work with the consortia, particularly the NAABB. 

NREL has had discussions with the NAABB sustainability team, primarily James Richardson, economist 

from Texas A&M. While the NREL models are developed from an engineer economics standpoint, the 

NAABB sustainability team is using exciting econometric tools that can further utilize the NREL work. 

While the NREL models are currently supplied by literature data, we look forward to incorporating 

research data from detailed bench and pilot scale experimentation. 





Page 47 of 223






Coordination with other efforts on LCAs and technoeconomic assessments should be exploited to 

maximize benefits. 


Little yet but project is young and shows excellent promise here. 


Models will be used to assist the DOE with target development and research interactions. 




The potential for technology transfer is very good. 


Not appropriately linked to consortium projects yet. 


Plans for publication and model sharing are very valuable. 


Again, integration with consortia is taking place as we speak. We look forward to this expanded 

collaboration with other tea and lca practioners. 






This seemed to be the project that was most on target with what the OBP program needs. Once again, any 

responsible technoeconomic modeling effort adds value. 


Strong project with good sensitivity analysis. 


Rigorous algae baseline technoeconomic analysis and user models have been developed for the Biomass 

Program and algae community - 3 pathways examined: open pond, photobioreactor, heterotrophic. This is 

a nice and low-cost techno-economic analysis project. Speaker needs to point his (laser) pointer to what 

he is talking about on the screen. Also, for the non-economists, please explain how to read the sensitivity 

X-Y plots. 

Page 48 of 223






Adaptable TEAs are important for assessing technological progress. However, several TEAs have been 

published recently and another is being developed, apparently independently, at INL (also reviewed). The 

major advance that must come from this project is a model that is easily used and modified by others. 

That should be the primary goal. "User-friendly" is mentioned in the presentation. For example, any use 

of dissolved air flotation and chemical coagulation is unrealistic and will need to be substituted for lowcost, 

low energy intensity technologies. 

The apparent involvement of undisclosed commercial partners raises a question about objectivity of the 

TEA assumptions. Presumably safeguards are in place, but these should be described. Of course, many of 

the OBP projects include commercial partner. The particular importance in this case is that a general 

decision support tool for the algae community is being developed. Transparency and ability to modify the 

assumptions should continue to be emphasized. 

For the heterotrophic model, the documentation should make clear that cellulosic sugar is not yet 

available. Use of cellulosic sugar is a huge assumption at this point. The projected autotrophic lipid 

productivity by both ponds and PBRs is reasonable for near term capabilities. The much higher 

evaporation and blowdown of the ponds compared to the PBRs should be substantiated--how was the 

evaporation of PBR cooling water estimated? 

The PBR sensitivity analysis is stretched beyond the conceivable, based on this calculation: 

The 8-cm diam. tubes contain 63 L of liquid per m2 of tube. The presentation gives 1.25 g/L-d as the base 

case productivity, or 63 L x 1.25 g/L-d = 78 g/m2-d. This is already a very high productivity, which is 

then projected in the sensitivity analysis to 3.0 g/L-d or 234 g/m2-d, which is not credible. The same 

comment could apply to the 60 g/m2-d projected for ponds. Sensitivity analysis might be conducted in 

terms of a combined modest improvement in productivity and in cost, to achieve more rational estimates. 

The tornado graph single factor graph is a step towards that. 

What if any efforts are being made to complement the apparently duplicative effort of INL (Newby 

presenter)? 


Very relevant project, good approach, and good presentation. 


Best of the modeling projects and well organized and an important effort. The PI should interact actively 

with the consortiums, however, to bring this model to appropriate execution. Would be very useful to use 

this model to "harness" the overal efforts of the consortiums, and to organize them towards a single goal 

of commercializing a specific pathway. 

Finally, be careful on the AD assumptions. An added carbon stream will be required to get the methane 

production if defatted algae is the principle feed. 


This project is likely the best investment made by DOE fro the algae platform. Flexibility should be 

incorporated into the model and the application should be be expanded and required for research efforts. 

Page 49 of 223




Care should be taken in presenting results as a tornado diagram (sensitivity diagram), because 1 factor at 

a time sensitivity analysis can be misleading. 


Project: 9.6.5.2 Argonne National Lab 

Title: Algae Life Cycle Assessment with GREET 

Presenter: Ed Frank 



Approach 7.29 0.70 7 

Progress 6.86 0.35 7 










The Approach has been quite good, based on the earlier work with the GREET model. Each additional 

study improves the work. 


This project is building an algal process model (APM) to extend GREET, which has been used for other 

energy sources (fossil fuels and others). Such a model is needed. 


– Using GREET ensures accurate, consistent comparison with previous and other OBP LCA assessments 

– Providing an algae LCA framework is an effective strategy to obtain comparable, transparent, 

reproducible analysis and to increase data accretion and availability. – Objectives are associated with 

metrics, tasks, and dates. 




The project goal is to create an LCA tool as well as to conduct a specific LCA for current schemes of 

algal biofuel production. The goals are technically sound and well defined. 


Page 50 of 223





Consistent approach with other GREET modeling. 


Thank you very much. We hope that with the GREET LCA framework, researchers and stakeholders will 

be able to evaluate algae pathways with other biofuel and transportation pathways consistently. 



management plan has met its objectives in achieving milestones and overcoming technical barriers 



Progress has been commensurate with the lifetime of the project. 


This study is 75% complete. Presentor cites difficulties with lack of validated data and the fact that some 

required data are "proprietary". Despite this, complete initial APM is in hand with an extended GREET. 


– The LCI inventory is diverse and open, covers many scenarios and a large fraction of the cumulative 

energy demand. – Early analysis results demonstrate ability to evaluate many scenarios. 




The project has met its milestones. A prototype was developed and described, a sensitivity analysis is 

underway and on time for summer 2011 deadline. 



Baseline scenarios near completion, but value of project is for evaluation of other scenarios. 


We agree that the project value will be increased by evaluating new scenarios. With completion of the 

technology options and scenarios currently under evaluation, we anticipate increased effort in this regard 

in the coming year. 





Page 51 of 223






The project is clearly relevant. The model should be exercised to provide cases for validation and use. 


This type of model is important to guide comparisons across projects. 


– The work addresses several MYPP barriers including St-D, At-A, At-C. – The project engages several 

national labs, OBP projects, and companies. 




The project is broadly relevant to platform goals. 



A valuable tool for down-selecting projects if data is available, if model has requried flexibility, and if the 

model is accompanied by techno-economic analysis. 


Flexibility for GREET design in general and algae pathways in particular has been a key consideration by 

ANL. We will continue to pursue flexibility and seek inputs from GREET users and the algae community 

in this regard. The tie to techno-economic analysis is important. In the coming year, with our LCA 

framework for algae in place, we hope to work with the NREL TEA team to define common parameters 

for both TEA and LCA and to exchange data and results between the two efforts. Similarly, working 

closely with experimental efforts will provide cases for use. 








assessments) 




end-use) 


Page 52 of 223






There was a good discussion of the CSFs. 


Demonstration of practicability of model needs additional work, including with data coming into 

literature from new projects and also older literature. 


– Risk factors were identified along with mitigation strategies – The project has identified a few critical 

issues for algae LCA 





There was a lack of criteria defining success at each unit operation, as well as information on the inputs 

and outputs of each module. It is important to have commercially realistic inputs and outputs for each unit 

operation. The PI should seek out people with real hands on experience growing microalgae at large scale, 

including harvesting, dewatering, and drying. 


Model acceptance and access to pilot data are critical. Project team is pursuing both. 


With the long history of GREET development and its large user basis, GREET is an excellent platform to 

develop consistent LCA and LCA comparisons. At the time of the presentation, ANL did not have 

definitive LCA results for algae yet, thus we did not go into detail how GREET and the APM are being 

used. With the completion of the APM, GREET expansion of algae pathways, and reports documenting 

the efforts and results over the summer 2011, practicality of the modeling capability will become 

apparent. We agree that use of data from processes that do not scale and that are not economically viable 

would not be meaningful for shedding light on future, large-scale algae operations. This is always a 

challenge for examining technologies that are not in commercial stage. Interactions with other analytic 

efforts such as those in NREL TEA team and other algae research efforts are always helpful to LCA 

efforts. Furthermore, with the APM, a few such processes, e.g., thermal drying, are in the inventory to 

allow people to quantitatively de-select them on an LCA basis and a few, e.g., centrifugation, are present 

because they may play a role if applied judiciously. Also, alternatives that are acceptable from an LCA 

perspective that are not viable economically, can suggest areas for technical development. That is, by 

providing a broader range of potential technology options for some critical algae stages, users of GREET, 

together with the APM, will be able to put their technologies in a broader perspective. 





Page 53 of 223






This aspect could be improved. There should be an attempt to integrate several of the modeling efforts, 

and there should be some movement towards defining best practices. 


The project personnel are working with Exxon and other algal biotech companies. 


– Algae LCA analyses will become systematic and comparable – The community will be able to address a 

larger number of scenarios 




The potential for technology transfer was good. The presenter emphasized extending analysis to 

approaches and developments by other research groups. 


This was undeveloped. Although a good approach, much of the assumptions and process pathway is 

rather devoid of issues described in the reports of Benneman and co-authors. Contact with the 

consortiums and groups (or those elements thereof that are actually growing and harvesting mciroalgae) 

woud be helpful. 


Publication and use with NAABB should be the first steps to broad acceptance and usage. 


We agree that defining best practices is important. Also important is developing a clear understanding of 

methodological uncertainties within LCA of algal fuels. Our strategy for addressing both of these is to 

facilitate consistent comparison amongst analyses so that analysts can identify and define best practices 

rather than being distracted by unintentional systematic differences in their models. ANL will make 

GREET and APM available in the public domain with technical publication. This plus community 

engagement is our vision of technology transfer. 






The projects on LCA add value. Using the models in predictive mode should be a future effort. 


This model is being competently built. 

Page 54 of 223





Too early to tell, as this is the initial stages of this project. 



LCA modeling is important and GREET is a common format. Because the algae biofuel process train is 

not well developed currently, not too much effort should be put into details at this point. However, the 

presentation did not provide much information on the scenarios that have been modeled. For example, 

what algae cultivation technologies have been implemented in the inventory? Is biomass transport 

included, etc.? 

The developers should strive to make the model transparent and easy to adapt/modify by outside users. It 

should be tied to the parallel TEA model. 


Very good project and presentation. 


This is a good first start, but may not produce a useful outcome unless grounded in the realities of 

commercial microaglae production. The PI would be well advised reading in detail the Benneman and coauthor 

reports, spending time at facilities that grow microaglae, and then re-reading these reports. The 

development of a Life Cycle Analysis after this activity would help develop better pathways and inputs 

and boundary conditions. Make sure all these assumptisns for each unit operation reflect the wisdom of 

Benneman/Oswald reports. 


The GREET modelling will help screen alternatives, but without an accompanying techno-economic 

model it has limited value. 


ANL learned a great deal from Benemann and other earlier work. The Benemann 1996 report was used 

extensively. Benemann, Weisman, Goebel, Oswald, and others developed many details, e.g., CO2 

transport and transfer, nitrogen recycling, and circulation velocity dependance upon algal species and size 

as well as economic constraints. They were helpful in establishing the structure and architecture of the 

APM tool. We feel that including the ability to model alternative scenarios and directly compare them 

unambiguously with the pragmatic earlier work is an expedient way to help the community to reach 

closure on some of the key issues. 

Project: 9.2.2.2 University of California - San Diego 

Title: Development of Renewable Biofuels Technology by Transcriptomic Analysis and Metabolic 

Engineering of Diatoms 

Presenter: Mark Hildebrand 



Approach 7.57 1.05 7 

Page 55 of 223




Progress 7.43 1.40 7 










The organization of the work seems to be very focused. Tasks are layed out in a very sequenced fashion. 

There are some questions that the investigator should address even for this exploratory project. Baseline 

oil production performance and how it compares to algae is one example. How does it fit with the OBP 

algae program and deliverables is another? Overall, this will be a contributory exploratory program. 


This project's goals are basic science aspects of the Roadmap to understand TAG production under silicon 

versus nitrogen limitation, understand genes involved in control points for carbon flux, and develop 

genetic tools (including transformation and knockdowns with RNAi, too) for manipulation in diatoms. 

Thalassiosira pseudonana and Cyclotella cryptica are the key targets and the PI suggests that what is 

learned in these systems can be transferred to most other diatom biofuels' targets. 


By characterizing gene expression changes using transcriptomics, fundamental aspects of carbon flux and 

partitioning can be elucidated, providing gene targets for metabolic engineering. 




The overall goal is to perform metabolic engineering of diatoms for higher TAG content. The specific 

goals are to compare transcriptional analyses of two diatoms, identify targets for engineering, and then 

engineer improved diatom strains. The science is very sound and was well presented. 


For what it is—a largely academic approach—it's a very solid approach. It lacks a basis for supporting 

realistic commercial growth of microalgae, i.e wild and native mixed cultures, but will provide very good 

scientific knowledge and advance our general understanding of diatom metabolism with respect to lipid 

biosynthesis. 


Although the approach is fundamental research, a baseline and commercial goals for productivity and 

yield would help calibrate the project. 

Page 56 of 223





The organization of the work seems to be very focused. Tasks are layed out in a very sequenced fashion. 

There are some questions that the investigator should address even for this exploratory project. Baseline 

oil production performance and how it compares to algae is one example. How does it fit with the OBP 

algae program is another. Overall, this will be a contributory exploratory program. 

Response: We are measuring increased performance based on the improvement of TAG production. 

Baseline data was presented in slides 9 and 11 of the presentation. The fit with OBP relates to the 

contribution of biomass and lipid accumulation as a component of an overall production system. Cost 

analyses indicate that improvement of these factors will be a major contributor to cost reduction. ----------- 

--------------------------------------------------------------------- 

This project's goals are basic science aspects of the Roadmap to understand TAG production under silicon 

versus nitrogen limitation, understand genes involved in control points for carbon flux, and develop 

genetic tools (including transformation and knockdowns with RNAi, too) for manipulation in diatoms. 

Thalassiosira pseudonana and Cyclotella cryptica are the key targets and the PI suggests that what is 

learned in these systems can be transferred to most other diatom biofuels' targets. 

No response. -------------------------------------------------------------------------------- 

By characterizing gene expression changes using transcriptomics, fundamental aspects of carbon flux and 

partitioning can be elucidated, providing gene targets for metabolic engineering. 

No response. 

-------------------------------------------------------------------------------- 

See Overall Impressions text. -------------------------------------------------------------------------------- 

The overall goal is to perform metabolic engineering of diatoms for higher TAG content. The specific 

goals are to compare transcriptional analyses of two diatoms, identify targets for engineering, and then 

engineer improved diatom strains. The science is very sound and was well presented. 

No response -------------------------------------------------------------------------------- 

For what it is—a largely academic approach—it's a very solid approach. It lacks a basis for supporting 

realistic commercial growth of microalgae, which wil be wild and native mixed cultures, but will provide 

very good scientific knowledge and advance our general understanding of diatom metabolism with 

respect to lipid biosynthesis. 

Response: As a first step, unialgal systems must be investigated, but knowledge could then be applied to 

more complex systems. The goals are to provide practical knowledge to benefit production systems – in 

this sense it is not strictly an academic approach. -------------------------------------------------------------------- 

------------ 

Although the approach is fundemental research, a baseline and commercial goals for productivity and 

yield would help calibrate the project. 

Page 57 of 223




Response: The funding amount has limited this to a laboratory-based study, but baseline productivity has 

been measured (slides 9 and 11 in the presentation) and provides a basis for monitoring improvement. To 

our knowledge, there is no existing data relating lab-based productivity to large-scale outdoor 

productivity in any study system (and this is likely to be highly variable depending on the species), 

therefore we cannot predict how our measure of improvement will translate to production. However, the 

only way to determine this is to generate the improved strains and test them in both scenarios. Based on 

first principles, it is logical to assume that at least some of the improvements generated in the lab will 

result in improved production. 






Progress appears to be acceptable at this stage. 


The project is about 40% complete. Excellent progress on knockdown tools (antisense and RNAi) and 

characterization of TAG under cell arrest by Si or N or MT inhibitors is evident. Transcriptomics of 

Cyclotella are advanced, and key enzymes related to carbon flux are being characterized with good 

progress. 


A whole-transcriptome analysis of TAG accumulation during silicon limitation in T. pseudonana is nearly 

complete, and samples are prepared for a similar analysis of silicon and nitrogen limitation in C. cryptica. 

Genetic manipulation tools have been developed. A clearer understanding of metabolic processes 

involved in TAG accumulation has resulted, and target genes have been identified for manipulation. 

Distinct stages during the TAG accumulation process have also been identified. 




The project has made progress in its objectives and seems to be on the way to meeting its milestones. The 

timelines for different milestones were not specifically stated. 


Very good work at an appropriate pace. 


Strong initial progress, but it need to be calibrated to real world performance and commercial needs 


Progress appears to be acceptable at this stage. ---------------------------------------------------------------------- 

---------- 

Page 58 of 223




The project is about 40% complete. Excellent progress on knockdown tools (antisense and RNAi) and 

characterization of TAG under cell arrest by Si or N or MT inhibitors is evident. Transcriptomics of 

Cyclotella are advanced, and key enzymes related to carbon flux are being characterized with good 

progress. 

No response. -------------------------------------------------------------------------------- 

A whole-transcriptome analysis of TAG accumulation during silicon limitation in T. pseudonana is nearly 

complete, and samples are prepared for a similar analysis of silicon and nitrogen limitation in C. cryptica. 

Genetic manipulation tools have been developed. A clearer understanding of metabolic processes 

involved in TAG accumulation has resulted, and target genes for manipulation. Distinct stages during the 

TAG accumulation process have also been identified. 

No response. -------------------------------------------------------------------------------- 


The project has made progress in its objectives and seems to be on the way to meeting its milestones. The 

timelines for different milestones were not specifically stated. 

Response: Yes, timelines were inadvertently omitted in the presentation, but are included in the proposal. 

-------------------------------------------------------------------------------- 

No problem here. Very good work at an appropriate pace. Quite competant. 

No response. -------------------------------------------------------------------------------- 

Strong intial progress, but it need to be calibrated to real world performance and comemrical needs 

Response: Yes, once improved strains are generated, that is the goal. 







One can see the relevance of microorganism production of lipid studies. The exact relevance of this to 

OBP program could be better delineated. What are the targets for this project? 


The basic science, including genomic tools being developed, have the potential to make a strong 

contribution to feedstock development for biofuels. 


Improving microalgal lipid accumulation characteristics will have a major impact on biofuels production. 

Page 59 of 223







Although the project has considered applications of its outputs, it is very unclear how relevant the results 

will be to large-scale applications. 


The key question is relevant to what? If its commercial production of microalgae under conditions that 

can be supported in open pond culture, at that scale, this work is not relevant. If the relevance is to basic 

scientific understanding of diatom metabolism with respect to lipid biosynthesis, then this is extremely 

relevant. 


Baseline and long term goals for strain performance would clarify and emphasize relevance. A method 

and partner for evaluating commercial potential needs to be identified. 


One can see the relevance of microorganism production of lipid studies. The exact relevance of this to 

OBP program could be better delineated. What are the targets for this project? 

Response: Targets are to develop procedures for improving strains (see slide 5 and 26). ---------------------- 

---------------------------------------------------------- 

The basic science, including genomic tools being developed, have the potential to make a strong 

contribution to feedstock development for biofuels. No response. ------------------------------------------------ 

-------------------------------- 

Improving microalgal lipid accumulation characteristics will have a major impact on biofuels production. 

Response: This is exactly the point for questions by other reviewers related to the relevance of the 

research. -------------------------------------------------------------------------------- 


Although the project has considered applications of its outputs, it is very unclear how relevant the results 

will be to large-scale applications. Response: We will never know the answer to that question unless we 

do the work and test the strains under production conditions. Cost analysis justifies the effort. -------------- 

------------------------------------------------------------------ 

The key question is relevant to what? If its commercial production of microalgae under conditions that 

can be supported in open pond culture, at that scale, the answer is that this work is not relevant. If the 

relevance is to basic scientific understanding of diatom metabolism with respect to lipid biosynthesis, 

then this is extremely relevant. 

Response: What is the basis for the conclusion that this will not work at scale? To our knowledge there 

are no studies demonstrating this. We will never know the answer to that question unless we do the work 

and test the strains under production conditions. That is the goal of this project. 

-------------------------------------------------------------------------------- 

Page 60 of 223




Baseline and long term goals for strain performance would clarify and emphasize relevance. A method 

and partner for evaluating commercial potential needs to be identified. 

Response: The funding amount has limited this to a laboratory-based study, but baseline productivity has 

been measured (slides 9 and 11 in the presentation) and provides a basis for monitoring improvement. To 

our knowledge, there is no existing data relating lab-based productivity to large-scale outdoor 

productivity (and this is likely to be highly variable depending on the species), therefore we cannot 

predict how our measure of improvement will translate to production, however, the only way to determine 

this is to generate the improved strains and test them, and based on first principles, it is logical to assume 

that at least some of the improvements generated in the lab will result in improved production. If 

additional funding can be provided to allow us to interface with a commercial partner, we would be happy 

to do so. 








assessments) 




end-use) 




As an exploratory program, some latitude can be given in expressing these. 


Demonstrating genetic manipulations and manipulations of nutrient "checkpoints" and "switches" that are 

persistent over many generations of diatoms is important to final success of this excellent basic scientific 

work to biofuels' development. Given that GMO algae may not be acceptable in commercial use in open 

facilities, characterizing heterotrophic growth in the context of the checkpoints/carbon flux under N and 

Si deprivation may be of interest----and trying to find natural switches for the cell cycle that can be 

applied w/o transformation. 

Given that diatoms reach a cardinal point where sexual reproduction is necessary to restore vegetative 

size, how will the genetic transformations and knockdown capability react to the diatom life history? Of 

course, some diatoms use vegetative processes to restore the largest cell size of the population, in the 

absence of sex, but as the population size decreases, sex is still critical in most species to restore cell size. 

Please reflect on these problems in your work. 

Page 61 of 223





Success relates to generating strains with improved TAG accumulation. Identifying genes that enable this 

without decreasing overall productivity is the challenge. 




The critical success factors were focused on identifying gene targets. Within this narrow biological focus, 

the project presentation included adequate plans to address these factors. 


In the context of understanding metabolic synthesis of lipids in diatoms, the PI has done an excellent job 

of laying out the appropriate pathways and metabolic markers. There is no doubt the data produce is and 

will be high quality. 


Critical success factors are clear focus on lipid production and accumulation in a real world setting 


Demonstrating genetic manipulations and manipulations of nutrient "checkpoints" and "switches" that are 

persistant over many generations of diatoms is important to final success of this excellent basic scientific 

work to biofuels' development. Given that GMO algae may not be acceptable in commercial use in open 

facilities, characterizing heterotrophic growth in the context of the checkpoints/carbon flux under N and 

Si deprivation may be of interest----and trying to find natural switches for the cell cycle that can be 

applied w/o transformation. 

Response: We appreciate the concern about GMOs, but at this point in the development of algal biofuels 

technology, it is not clear whether or not GMOs will be allowed. If we assume the need for GMOs in a 

biofuels setting will be as great as is the widespread use of GMOs in food crops and approval for their use 

is gained, then this research could directly translate into production strains. If not, then it will still provide 

a fundamental basis for understanding that will inform approaches for selection of desirable traits 

(potentially using mutagenesis). Regarding heterotrophic growth, we are examining that aspect in another 

project (C. cryptica has marginal heterotrophic growth capability). Natural switches for cell cycle arrest 

are Si and N limitation. 

Given that diatoms reach a cardinal point where sexual reproduction is necessary to restore vegetative 

size, how will the genetic transformations and knockdown capability react to the diatom life history? Of 

course, some diatoms use vegetative processes to restore the largest cell size of the population, in the 

absence of sex, but as the population size decreases, sex is still critical in most species to restore cell size. 

Please reflect on these problems in your work. 

Response: The reduction in size phenomenon is not universal for diatoms, only those with a higher level 

of silicification than average in which the cell wall is highly rigid follow this life cycle process. Many 

species do not reduce in size with generation, and it is actually extremely difficult to induce the sexual 

cycle in them – they have a strong preference for vegetative reproduction. Our lab has published on the 

size reduction question, and provided direct proof that it need not happen (Hildebrand et al. 2007, J. 

Phycol. 43, 730–740), and other publications have documented the same. Of particular relevance is that 

the species we are focusing on do not undergo size reduction and triggering of the sexual cycle, and many 

Page 62 of 223




of the other species we are examining have a similar feature. The sexual cycle could potentially be taken 

advantage of (if it was easy to reproducibly induce) because cells would go through a haploid stage which 

may facilitate genetic or transgenic approaches to improve productivity. 

Success relates to generating strains with improved TAG accumulation. Identifying genes that enable this 

without decreasing overall productivity is the challenge. 

Response: We believe that identifying and testing multiple gene targets will facilitate achieving that goal. 

See Overall Impressions text. The critical success factors were focused on identifying gene targets. 

Within this narrow biological focus, the project presentation included adequate plans to address these 

factors. 

No response. In the context of understanding metabolic synthesis of lipids in diatoms, the PI had done an 

excellent job of laying out the appropriate pathways and metabolic markers. There is no doubt the data 

produce is and will be high quality. 

No response. Critical success factors are clear focus on lipid production and accumulation in a real world 

setting 

Response: That is the ultimate goal. 







I would like to see this developed in greater detail. There is no doubt that publications will be 

forthcoming. 


Continued publication and presentations and interactions with more applied groups in the Biofuels' 

portfolio are important to transfer scientific advances made here into practical advances toward biofuel 

goals. 


Future work will be to test the effect of manipulation of specific genes on TAG accumulation. When 

successful, the technology of these modifications can be transferred to production systems. 




Technology transfer will be primarily publications and presentations. 


This is mostly a single PI project. The partner identified (Dr. Allen) is appropriate. 

Page 63 of 223





It was not clear how resutls would be shared or how any potential commercial opportunity would be 

evaluated. 


I would like to see this developed in greater detail. There is no doubt that publications will be 

forthcoming. Response: In addition to publications and presentations, the research will develop our 

understanding of the fundamental biological control processes over lipid and biomass accumulation. 

Given the fundamental need for this in the context of a production system, it is likely to benefit other 

aspects of the Biomass Program research. ---------------------------------------------------------------------------- 

---- 

Continued publication and presentations and interactions with more applied groups in the Biofuels' 

portfolio is important to transfer scientific advances made here into practical advances toward biofuel 

goals. 

Response: Agree. 

-------------------------------------------------------------------------------- 

Future work will be to test the effect of manipulation of specific genes on TAG accumulation. If 

successful, the technology of these modifications can be transferred to production systems. 

No response. -------------------------------------------------------------------------------- 


Technology transfer will be primarily publications and presentations. Response: Initially yes, but if better 

performing strains are produced then material for testing in large scale systems will be available. ---------- 

---------------------------------------------------------------------- 

This is mostly a single PI project. The partner identified (Dr. Allen) is fine. However, there is no real 

issue here, given the projects objectives and capacity of the PI. 

No response. -------------------------------------------------------------------------------- 

It was not clear how resutls would be shared and any potential commercial opportunity would be 

evaluated. 

Response: Results would be shared via publications and presentations, and if future funding was 

available, or if a commercial partner became interested, large scale testing could be pursued. 




Page 64 of 223






As a project generating science, I like this work. It may even be a supporting project for the OBP efforts. 

However, it would be better to spell these expectations out to make the case. Perhaps this would have 

been better funded totally by NSF? 


Collaboration with Venter Institute a plus. This project is making excellent progress in the area of 

understanding the lipidome of diatoms and providing tools and metabolic data to manipulate it. 


PI is well-versed in the art of cell and molecular biology of Thalassiosira and Cyclotella. Useful results 

will emanate from this work. The only caution is that changes in transcript mRNA level often are not 

reflected in changes in protein amount or activity. How is the PI planning to account / check for the 

possibility that transcriptomics are not always the best approach to assess flux and metabolomics?? 



A model presentation of theory, hypotheses, and data. The team has significant accomplishments at 13 

months into the contract, while using among the most modest budgets of all projects reviewed. The means 

to provide affordable silica to diatom production is practical issue to address if they are to be used in 

biofuel production. 


Within the area of fundamental research for algal biofuels, this is a very good project and was well 

presented. 


This work is high level academic work, with the ever constant promise that the work will support the 

commercial effort. That said the work is limited to themetabolic engineering of a single strain that will 

require conditions of sterility at large scale deployment. As such, this project may be more suited in DOE 

Basic Sciences or NSF. 


Technically strong work, but the presentation did not address how research gains would be evaluated for 

commercial value, the magnitude of improvement that is required and expected, and on what time frame 

this might occur. 


Project: 9.2.1.2 University of Georgia 

Title: Improving cost effectiveness of algae-lipid production through advances in nutrient delivery and 

processing systems 

Presenter: K.C. Das 


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Approach 4.29 1.83 7 

Progress 4.14 1.36 7 










Many different tasks have been proposed for this project. In addition, many others were added as various 

tasks did not work out. This is not scientifically structured work. 


The focus of this project is to deliver nutrients to algae via poultry litter (feces) and to explore uses of 

residual biomass. It appears that initial work may have been spread over more different goals than 

optimal. 


Project addresses key targets in Biomass Multi-year Program Plan related to cost competitiveness of 

biomass derived biofuels 




Approach is very multi-faceted with a wastewater utilization theme. Approach and results were all 

laboratory scale. Unfortunately, it was hard to understand many of the logical connections between the 

different project themes. 


The PI is addressing a real world concern and in a manner that is keeping an eye on the realities of 

commercialization. The PI could focus down a bit more and aggressively pursue a few stragetic pathways. 


The project goals should be defined around specific economic and productivity targets. The potential 

magnitude of the nutrient opportunity should be defined in terms of potential tons/year of biomass. 




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It is clear that results have been taken, mainly in an exploratory mode. 


The project is about 85% complete and has involved many departments at UGA and one partner in 

Auburn. The project has been unable to use poultry litter to supply nutrients to algae (e.g. Chlorella) and 

produce biofuel-quality levels of lipid; this is due to the high N causing biomass growth rather than lipid 

accumulation (a relatively well known fact in background literature). Consequently the project has 

focussed on biomass production, but the nutrient solution is dark so algae have to be grown in shallow 

raceways and the product is smelly. 


Investigators are developing sustainable, cost competitive, algae technologies to enable biofuel 

production and creation of a new domestic industry. 




Unfortunately, it was difficult to evaluate progress, stated objectives, and milestones 


The data presented represents reasonable progress. Oveall the data lacked depth and the work seemed to 

move from concept to concept, with only semi-superficial depth to the data accumulated. Tone should 

nail down a pathway and really work to undestand it at all levels. 


The dark color of the PLE, the reduced productivity of growth in raceways with PLE, the odor issue, and 

water recycle problems with possible concentration of color bodies indicate that this is not a viable 

nutrient source. 







The work is so scattered it is hard to credit much true relevance to the OBP program goals. 


Nutrient supply is an important part of production of algal biofuels; however, the dark extract and 

odiferous product may limit applications. 


Consistent with the objectives of the Algal Biomass program, although part of the emphasis is the cleanup 

of poultry-derived wastewater. 

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It was very unclear how these results are relevant to program platform. 


Although a very good project, given the objectives, these objectives are not focussed on large scale fuel 

production. The project does address a solid waste stream that needs to be managed, and how to do it with 

postiive energy economics. Microalgae a way to treat waste water is very useful. 


There appear to be many issues with PLE that make the nutrient source problematic for algal biofuel 

production: color, odor, reduction of productivity. 

The economics and application at scale of immoblized nitrogen fixation should be presented. 








assessments) 




end-use) 




Some set of CSFs should have been devised, even though the project was exploratory in nature. Industries 

are faced with this same task for their exploratory projects. Books have been written on how to control 

and gauge this work. Some sort of system should have been employed here. 


The project is nearly complete a different approach to nutrient supply for algal biofuel's may be required 

based on its results to date. 


Poultry litter needs to be shown as a potential low cost nutrient source for algaculture. Advanced fiber 

flocculation needs to be established as a promising technology for harvesting. 



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Unfortunately, there was not enough detail presented to adequately evaluate success factors. 


The work presented addressed a waste stream, poultry litter, and a means (microalgae production) as a 

means (among others) to treat it. THis was useful but there appeared to lack clear and comprehensive 

trigger points for decision making. 


The project success factors do not appear to be defined in terms of technical and economic hurdles to 

algal biofuel production. 







It is unclear whether there were any outputs that could be useful. 


This project's results are somewhat cautionary and should be shared with other DoE projects. 





It was unclear whether and how the results of these disparate lines of laboratory research would benefit 

the Biomass Program. 


This was not well developed. 


If research results and theory indicate that the use of PLE is not a viable option for algal biofuel 

production, valuable informatton sharing would be to reach these conclusions and explain the nutrient 

charactorstics that are unsuitable. 




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This work was too scattered, and it is unclear what the outcomes were. Maybe there are some buried 

nuggets but they will remain hidden I believe. 


The project investigated a number of things (including cyanobacteria) in addition to the main goals, and 

more focus on solving the extract's problems (coloration, odor) might have been in hindsight, better. It 

would be difficult to recommend that other DoE funded projects take this route to solving the critical 

problems of nutrients supply based on the data that were presented. 


This project has the potential of both cleaning up wastewater and generating useful algal biomass. But I 

would not feed such microalgal biomass back to poultry. 



Goals and research topics are too numerous and scattered, and the results are not presented on all topics 

mentioned: 

Cultivation-related: poultry waste media, N-fixers, CO2 capture 

Processing-related: coagulation with fibers, cell disruption, lipid extraction, residuals for feed 

Other activities: ponds vs PBRs, ethanol production, carpet industry wastewater media 

Poultry waste media were very dark with only 1% poultry waste. Growth results are difficult to judge 

from single data graph shown. 

Ponds vs. PBR: setup is not ideal because the ponds are shallow in deep tanks, causing shading. The 

productivity values for the PBRs are probably not corrected for lack of shading (edge effect), which is 

unrealistic at full scale. 

Carbonation column needs to be evaluated in terms of energy consumption, not only CO2 dissolution 

efficiency. The column energy consumption will be large. 

Only one graph and no conclusions provided for N-fixation experiment. Purpose not clearly stated. 

About seven papers are listed that appear to stem from DOE funding. Despite this nice output, the 

presentation lacked cohesion and few if any clear conclusions can be drawn from the result presented. 

The focus on wastewaters is potentially useful. 


The approach is all lab-scale based and multifaceted. It was unclear whether there was sufficient depth to 

the different lines of research to prove useful to the Biomass Program. 


This was a good project addressing an important waste stream. Well situated within farming industry, or 

solid waste management. 

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It appears that research results and PLE physical characteristics indicate that this in not a good nutreint 

source for algal biofuel production. If this conclusion is true, than the value to the algal biofuel program is 

an analysis of why PLE is unsuitable. The concept of immoblized nitrogen fixation need to be well 

defined in terms of commercial potential of algal biofuels. 

Project: 9.2.2.1 University of Maine 

Title: Production of higher alcohols liquid biofuel via acidogenic digestion and chemical upgrading of 

industrial biomass streams. 

Presenter: Peter van Walsum 



Approach 6.20 1.47 5 

Progress 6.20 0.98 5 










There is not a good description of the approach. There is no rationale detailed for us to follow the plans. 

When chemicals are to be the products rather than fuels, some more detailed discussion of the valuable or 

less valuable products is expected, and strategies for maximizing higher value products should have been 

part of the Approach. 


No breakthroughs needed. Project is focusing on robust technology, cheap feedstock, existing industries 

to achieve its potential. 


The approach was to use conventional wastes from existing local industries, evaluate three different 

pathways, and to develop process models. The research and approach were sound and well presented. 


Good and solid approach to upgrading via three reasonable pathways. Appropriate go or no go criteria, 

and is an appropriate pathway from microalgae biomass to a fuel. 


Good use of techno economic modeling. Project would benefit from clearer definition and 

communication of goals for the potential volume of fuel production and targets for capital and total fuel 

costs. 

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There is not a good description of the Approach. There is no rationale detailed for us to follow the plans. 

When chemicals are to be the products rather than fuels, some more detailed discussion of the valuable or 

less valuable products is expected, and strategies for maximizing higher value products should have been 

part of the Approach. 

Chemical prices are difficult to determine, and are usually quite volatile. Since this is a preliminary 

investigation, we are not focusing efforts on optimizing production of particular products. Generally, we 

have seen prices for most of the likely commodity type products (organic acids, ketones, esters) to be 

roughly in the same range, which is about 2-3 X the value of a chemically comparable fuel. Also, the 

value of any chemical will depend on its purity, and this introduces another layer of modeling and cost 

calculations to determine best candidates for purified chemical markets. 

Good use of techno economic modeling. Project would benefit from clearer definition and 

communication of goals for the potential volume of fuel production and targets for capital and total fuel 

costs. 

I agree that more clarity on volume of production and anticipated costs would be helpful. We will clarify 

these issues as we continue forward. 



management plan has met its objectives in achieving milestones and overcoming technical barriers 



Progress has been made as the project aims toward completion. More details and data could have been 

helful. 


Demonstrated fermentation and downstream unit operations, still need to demonstrate integrated process 


The presentation showed accomplishment of key milestones and progress--looks very good. 


Well done. Clearly a good pace of data that has been well taken, presented, and characterized. 


Successful use of pilot data in techno economic modeling. Modeling results may indicate that this process 

is not economically viable. 


Successful use of pilot data in techno-economic modeling. Modeling results may indicate that this process 

is not economically viable. 

The scale of these potential applications makes the economics challenging. However, they are 

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encouraging results if reasonable scale up could be accomplished, perhaps by tapping other available 

feedstocks available in particular locations. 







There is moderate relevance in that data on products and productivity will be forthcoming. Some of the 

products are not as valuable as projected, and conclusions should be provided in a controlled manner. 

Several details of how the experiments were conducted, and the lack of appreciation for some of the 

details argue for review beforehand. One example was the specific comments on the addition of 

significant levels of manure which contain organic acids, and the lack of correction for them. 


This project entails efforts at demonstrating low hydrolysis and fermentation costs, as well as an 

integrated bio and thermochemical platform. 


Excellent example of use of local resources for alcohol production, with potential use in other 

coastal/forested areas. However, it is unclear that this type of local resource utilization project could be 

scaled to meet significant transportation needs. 


The final products are appropriate fuels for the DOE Program. It’s good to look at other products than just 

biodiesel. 


It appaears that the potential fuel volumes from this process are very small. The mixed alcohols produced 

may have unique charactoristics which, combined with the small volumes, make it a nuiscence for fuel 

production. 


There is moderate relevance in that data on products and productivity will be forthcoming. Some of the 

products are not as valuable as projected, and conclusions should be provided in a controlled manner. 

Several details of how the experiments were carried out, and the lack of appreciation for some of the 

details argue for review beforehand. Specific comments on the addition of significant levels of manure 

with organic acids, and the lack of correction for them is an example. Details on experimental methods 

are documented in our quarterly reports. Time constraints of the presentation precluded giving many 

details on procedures. I checked back with my research student and he confirmed that, indeed as shown in 

the fermentation time plot, the initial acid concentration in the fermentation was near zero, indicating very 

little initial contribution of organic acids from the manure. The manure may have been free of acids 

because it was dried and acids originally present may have volatilized prior to the manure addition to the 

fermentation. 

Excellent example of use of local resources for alcohol production with potential use in other 

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coastal/forested areas. However, it is unclear that this type of local resource utilization project could be 

scaled to meet significant transportation needs. 

The resources under investigation in this project are too limited to provide significant volume of product 

for transportation. Several of such biomass sources would be needed to produce even a modest amount of 

fuel. However, these biomass resources are available and already alkali pretreated, thus they make for a 

good starting place to test and demonstrate the conversion pathways. 

It appears that the potential fuel volumes form this process are very small. The mixed alcohols produced 

may have unique characteristics which, combined with the small volumes, make it a nuisance for fuel 

production. Certainly the volumes we are looking at are small and they have value more as demonstration 

feedstocks than commercial opportunities. The nuisance is important. I think that if fuels are the final 

target then hydrotreatment of the mixed alcohols to gasoline range hydrocarbons (which has been 

demonstrated) would be the most likley way to go to market. Such hydrotreatment was not within the 

scope of this project. 








assessments) 




end-use) 




There is an extensive list of CSFs and how they are addressed. Challenges are also tabulated and 

discussed. 


Higher fermentation concentrations needed. 


Demonstrated completion of critical success factors and technology transfer--seems to have addressed 

them well. 


I really liked the net energy balances that were calculated. A lot of time was devoted to the factors being 

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evaluated, and the PI had reasonably targeted appropriate criteria, from fermentation titres and yields 

thorugh legal regulation. The energy balance, however, was most appreciated. 


Potentially low volume of fuels combined with the end-use of mixed alcohol might be additonal success 

factors to be addressed. 


Higher fermentation concentrations needed. Agreed. 

Potentially low volume of fuels combined with the end-use of mixed alcohol might be additonal success 

factors to be addressed. Agreed. Scale would need to be increased for commercial operation. Mixed 

alcohols may need to be hydrotreated to gasoline hydrocarbon. 







It is not easily determined if the deliverables and conclusions are of value to the OBP program or not. 

There are many details that would need evaluating to ultimately determine the value of the outcomes. It is 

believed that publications will be an output. 


Working closely with OTFF. FMC option looks less promising (low titer). Terrabon relationship is 

developing. 


Collaboration and technology transfer were still in process. 


I did not get a strong feel for this. 


Widespread sharing through many publications. 


It is not easily determined if the deliverables and conclusions are of value to the OBP program or not. 

There are many details that would need evaluating to ultimately determine the value of the outcomes. It is 

believed that publications will be an output. Publications (non-peer reviewed) have been produced. 

Manuscripts are in preparation for submission to peer reviewed publications. 

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There shold be more comfort in the project and its outcomes than I register. There seem to be many 

details that one needs to know before one could nod in agreement. I fear that more experiments would be 

the more likely result. The production of light organic acids may not be a valuable outcome. However, 

this may just set the stage for a separate project beyond this. 


A main objective of this effort is to achieve low cost enzymatic hydrolysis and complete carbohydrate 

fermentation of biomass feedstock. - What are the cellular biochemical targets of the purported 

fermentation? - What are the products to be developed by this approach? These specifics are a must for 

the proper evaluation of the project. 


Very nice presentation of a local resource utilization project. 


Overall a good project that is well done. Solid pathways considered. Looking at three pathways? What are 

the criteria for comparison? Seaweed digestion. Ends up a dirty liquid (pigments et al). How does one 

purify the fuel and how does one dispose of the fermentation media? What is the go no go criteria? How 

do alcohol yields compare against simple AD of chicken manure adn methane gas produciton (and on site 

combustion?) 


My strongest impression is that the potential fuel volumes are too low and too dispersed geographically to 

be a viable transporation fuel source. The capital and production cost estimates indicate that this 

technology has very big hurdles for economic viability. 


I think that the “comfort” issue may be that much of the project has been working to down select from 

three to one preferred process pathway. The initial investigations were done at a preliminary level. We are 

currently focusing on the ketonization pathway and I believe that the data we are generating will help to 

develop a higher level of comfort. Now that we have selected the ketonization route as the preferred 

pathway, our final product is primarily isopropyl alcohol along other secondary alcohols. Optimally we 

would like to see higher proportions of longer chain alcohols, as these are more compatible with gasoline 

infrastructure. This in turn requires fermentation s that accumulate larger quantities of butyrate and other 

longer chain acids. 

Criteria of comparison between the three pathways were yield, energy balance, cost and ease of 

implementation. The ketonization pathway won out on energy balance and ease of implementation. Ease 

of implementation was critical in order to make progress on the time line we had set for ourselves. The 

seaweed digestion does turn dark. So far, we have found that purifying the fermentation broths is not 

necessary when using the ketonization route. The other organic materials from the fermentation media 

that accompany the carboxylate salts into the ketonization reaction appear to contribute to the chemistry 

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of deoxygenation. Converting unpurified carboxylate salts does result in a much greater variety of organic 

compounds, but these may not be problematic if the mixture is ultimately hydrotreated and blended with 

petroleum-based fuels. Thus, the fermentation media is disposed of by evaporating out the water and the 

solids are sent to ketonization. The ash from the ketonization contains CaCO3 and other alkaline 

components which would be recycled back to neutralize the fermentation step. The ash recycle would 

eventually require purging and it may have value as an alkaline soil amendment ( this is currently how the 

algae sludges are disposed of—as alkaline soil conditioner) A study of comparing methanogenic and 

acidogenic digestion of biomass is underway. Preliminary findings (not a part of this project) appear to 

suggest that the higher value of liquid fuels and chemicals can more than offset the greater complexity of 

the processing. However, AD does benefit from much easier scale down to process small volume 

resources, with the small-scale resources investigated in this study, this is likely to be true. We are 

finding, though, that at these smaller scales, the costs appear attractive compared to other liquid biofuel 

technologies. 

Project: 9.2.1.1 Montana State University 

Title: Extremophilic Microalgae: Advanced Lipid and Biomass Production for Biofuels and 

Bioproducts 

Presenter: Brent Peyton 



Approach 6.00 1.07 7 

Progress 5.86 1.12 7 










The project has proceeded in a logical fashion and has therefore had a reasonable approach. The next 

stage is to scale-up out of the lab, and this is what is planned. 


A major focus of this project is to screen algae that may be good lipid producers for growth at high pH, 

where they may be protected from competition and predators. 


Effort focuses on characterizing and quantifying alkaliphilic algal growth and lipid production for scaleup 

to open ponds. Issues to be addressed: -Help stabilize algal pond systems without genetic modification, 

-Increase CO2 uptake rates -Utilize low quality water -Provide TAG production rate and extent data in 

outdoor systems 

Page 77 of 223







The broad objectives are to develop and use high lipid producing alkaliphilic (sp?) strains from culture 

collections, determine kinetic parameters in lab-scale studies, and then test strains in open ponds at pilot 

scale at Utah State Univ. The science is sound and objectives are reasonable. 


It is relevant to approach production in extrememe environments for all the reasons stated. And the use of 

high pH water is reasonable but the presentation lacked a clear demonstration of just how much of this 

kind of water there is. What is the resource and what is the rational amount of fuel that can be produced? 

Where are these waters located (i.e., in cold climates open culture or photobioreactors are severely 

challenged). 

I think overall this is an interesting project, extreme environments for high pH water) that will be stressed 

in terms of resource allocation and commercial application. 

The PI should do more than just propose extreme pH. The defense of this approach, from a resource 

management and commercial application point of view needs to be defended. 

Also, present biomass data as gdw l-1. 


The project needs specifc goals for productivity and yield based on commercial algal biofuel production 

requirements. 


There is a significant amount of non-potable high pH and salinity water in the west, where potable water 

is extremely limited. Other, funded specific projects are focused on determining the water resources 

available across the country to determine where our work will be feasible. 






Progress has been good through the several years of work. Still, the large scale-up effort remains. 


The project is 65% complete and is now concentrating on characterization of 4 green algae and 4 diatoms 

but has found 30 strains that will grow at pH greater than 9.5 despite not being able to use carbonate as a 

C source (at least the main targets do not). 

Lipid production from these is high, based on Nile red; some analytical chemistry should be used to 

support the Nile red results. 

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This project's research on feedstock appropriate to high pH waters may be particularly germane to the 

West where water is limiting. 


(1) Demonstrated high lipid production with alkaliphilic algae including Scenedesmus WC-1 and diatom 

RGd-1. (2) Significantly increased growth rates and lipid yields with different culturing conditions (pH, 

aeration, light:cycling, bicarbonate) (3) Filed provisional patent on bicarbonate work (jointly supported 

by DOD and AFOSR) 




Lab scale experiments are completed, will address 50 gallon and 500 gallon outdoor open pond studies in 

summer of 2011. 


Much of the data is focussed on calibraiton curves (or related thereof) for Nile Red and lipid density. Past 

that, there appears to be some shake flask work that looks at batch cell growth and some ntrate uptake, but 

past that this work is premature. The open pond data appeared to be one time only, over 24 days (almost a 

batch curve). This data was not well discussed or explained. 


Conceptual definition of a high pH production system is needed to put progress into commerical context. 


Open pond tests are scheduled for this summer. 







Relevance is still somewhat open to discussion. True it is algae, but the alkalophilic nature is special case 

until further development work is done. 


May be particularly relevant to algal biofuels' development in the West. Central premise of study remains 

to be tested, however (that high pH growing conditions are protective to algae vis-a-vis competitors and 

predators). 


Algal Biofuels have potential to produce significant amounts of biodiesel, but this is in the early stages of 

development. 

Page 79 of 223








There was little justification for the use of extreme cultures (to pH) in terms of the Biomass Program 

goals of large scale cultivation. What is the resource of this kind of water? Where is it found, and what is 

the opportunity to grow under these conditions? Very little data other than calibration curves for Nile Red 

was shown, so there is little real contribution to basic science yet. Overall, its commercial application is 

as of yet poorly defended. 


Preliminary techno economic modeling of a conceptual alakaline cultivation and harvest system and 

identification of possible geographies are needed to define relevance. 


We strongly disagree with the very negative comment here. This project has so far led to a provisional 

patent and a peer-reviewed publication with at least two to three others in the works. 








assessments) 




end-use) 




I interpret the CSFs presented as reasonable assessments. 


It would be good to close the loop on the contract by studying competitors and predators (including 

pathogens) that develop/are present under the high pH growing conditions and when algae are being 

grown up for biomass vs lipid accumulation. 


1. High pH conditions help maintain the stability of the culture and lipid productivity. 2. Organisms 

adapted to low value alkaline/saline conditions. 3. Outdoor tests show competitive rates and total yields of 

TAG accumulation. 

Page 80 of 223








The open pond growth data is over 24 days. That's a long time. How does this compare to commercial 

standard biomass productivities, and lipid production thereof. There was little comparison of data to a 

baseline expectation of any production level that would support scale up. 


Definition of a viable alkaline cutlivation system is critical to success. 


We will be defining a best case alkaline system based on the open pond tests in summer 2011. 







This aspect could be improved. After the larger scale raceway experiments, there should be some 

exposure to the OBP group at large. Publications have been forthcoming. 


Publications and presentations are being made at good level. The work yet to be accomplished will 

determine how successful tech transfer can be. 


1. Signed NDA’s with a number of companies on the bicarbonate work and are in license discussions 

with a large company in this area. 2. Considering patenting strains 3. Peer reviewed publications 4. Task 3 

Outdoor open pond tests at USU 




A patent application has been submitted applied for the bicarbonate results. However, no commercial 

partners have been identified. 


I did not see much other than a reference to University Partners. 


many of the listed publications in the presentation slides are not directly related to this work. 

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We are in negotiations with commercial partners on the provisional patent, but prefer not to name any at 

this time. 






Overall, the project has been successful at the research scale. The larger scale experiments will add 

significantly to the weight of the results. 


This is a useful study of algal biofuels' feedstock development for high pH waters; additional 

characterization of growing conditions relative to competitors, predators and pathogens will be important. 


More specific information is needed from this project: What is the rate of growth of the alkaliphilic algae 

(pH > 9.5), compared to neutral pH algae? Goal of the project is to achieve lipid composition >15%. 

What is the pH optima of the strains examined? In lab experiments, buffer systems are used (expensive). 

How is this going to be done in scale-up? 



The Goal Statement and Project Objectives have a mismatch that should be ironed-out. The Goal 

Statement and Critical Success Factors give the impression that alkaliphilic culture resistance to invading 

algae/pests is a hypothesis needing testing. However, the Project Objectives are all focused on lipid 

production. 

Extreme alkaline conditions would presumably have a metabolic cost to the algae leading to lower 

productivity than neutral pH waters. (e.g., Spirulina's relatively low productivity). Although, this need not 

be a major point of research, including some control non-alkaliphilic cultures in the lab and field 

experiments would provide direct information on this concern. Slide 7 may show this kind of experiment, 

but it is not clear. Controls could also be an opportunity to test the invader question raised by the Goal 

Statement. 

The presentation of the data is confusing in some cases. In Slide 11, what was the pH on the unbuffered 

culture? Slide 12: What does this mean: "No pH induced and low nitrate depletion TAG accumulation"? 

The effect of bicarbonate addition on lipid content is interesting, but what are the next steps to discover 

the mechanisms of the bicarbonate effect? Is it a pH effect or a carbon supply effect? Were the algae Climited 

prior to the bicarbonate addition? Are there any practical ways to implement this effect at large 

scale? 

A great strength is the use of outdoor ponds to scale-up, but unfortunately, this work has apparently not 

yet begun and so will be only short-term. 

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What is national availability of alkaline waters for this process? Include at least a preliminary resource 

assessment in this research. Although not a unique problem to alkaline groundwaters, the reporting should 

acknowledge difficulties with disposal of blowdown potentially contaminated with natural toxicants-metals, 

metalloids. 


Nice presentation of a nearly complete project. The impact of the project is uncertain until they have 

results from large-scale outdoor trials. 


Overall, this is a limited project (so far) academically and its commercial relevance is more suggestion 

than reality. No real case was made for the source of water (oveall amount, access, location...etc) other 

than to say it’s good to use unwanted water (which I agree with). Also, the point regarding a mechanism 

against contamination was relevant but poorly developed in the experimental approach and no data really 

addressed it. 


The project would benefit from development of a conceptual alkaline produciton system and definition of 

produdtivity and yield requirements to make the system viable for biofuel production. 


In the time allotted and the very defined format, it was difficult to present enough detail for a complete 

understanding of our results. These results are presented in the following publication, which is available 

online, and in print shortly: Gardner, R, Peters P, Peyton BM and Cooksey K. Medium pH and Nitrate 

Concentration Effects on Accumulation of Triacylglycerol in Two Members of the Chlorophyta. In Press. 

J. Appl. Phycology, 2011. 

Discovering the fundamental mechanisms behind the bicarbonate are not part of the proposed workscope 

for this much more applied project. Further, examining the susceptibility for pond contamination is part of 

this summer's work. We anticipate that the energy balances for our cultures/systems will not be 

significantly different than any other culturing systems. Detailed energy balances were not part of this 

organism-focused scope of work. 

Project: 9.6.1.3 Pacific Northwest National Laboratory 

Title: Macroalgae GIS Analysis 

Presenter: Guri Roesijadi 



Approach 3.57 1.18 7 

Progress 3.14 1.25 7 



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The Approach section suffers from a real baseline assessment of the logic for this tool development. 

There is a statement that macroalgae will be a feedstock of substance, but of course there is no back-up or 

consideration of the difficulty of harvest, transport and conversion of this feedstock compared to landbased 

feedstocks. 


This is a modeling project and the number of ciritcal variable included should be expanded. It is likely 

that the project would benefit from a detailed review of older and current publication on macroalgal 

production. 


GIS tools will be developed to identify cultivation sites with minimal competing impacts on existing uses 

of marine and coastal environments. 




The ultimate goal is to evaluate conversion of seaweed to alcohols. As a first step, the approach of this 

work is to conduct strategic analysis of US macroalgae production as feedstock resource. 



Inclusion of current information from the exisitng kelp industry and a sustainabilty analysis might make 

the production potential analysis better. 


1.1. Although the biomass potential of macroalgae as a biofuels feedstock in the United States is high, the 

resource is undeveloped. Whether it will be a “feedstock of substance” is not known. A national 

assessment of the production potential of macroalgae in U.S. waters is needed. Our year 1 deliverable, the 

FY10 report “Macroalgae as a Biomass Feedstock: A Preliminary Analysis” is an initial analysis of the 

resource, economics, and environmental impacts. This assessment was “high-level” due to the lack of an 

existing national macroalgae production industry. We conducted a preliminary analysis based on best 

available data, and, where appropriate, application of our understanding of processing other feedstocks to 

macroalgae. Cultivation of macroalgae as feedstock would not compete with an existing domestic supply 

chain or with available land, thus, avoiding conflicts faced by other feedstocks. It was beyond the scope 

of the program review to present the findings in detail due to time and format constraints. For the program 

review we presented key information from this report, meant to provide background for our resource 

assessment in FY11, which focuses on the question, “Where can this biomass be produced?” In the initial 

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analysis, we estimated a need for 32.3 million dry metric tons grown on about 11,000 km2 to meet 

nominal biofuels targets (i.e., a 1% gasoline replacement target). Additionally, we noted 

recommendations from the Pew Oceans Commission that marine aquaculture needs to move to offshore 

waters due to competition with existing uses or restrictions in nearshore marine environments. 

Developing the tool for locating production sites in U.S. waters, which do not conflict with existing uses 

or regulations, was considered a first-order need. Quantifying production potential and identifying 

potential cultivation sites are critical for our objectives and, after a baseline has been established, a 

comparison to other feedstocks will be useful. Other considerations related to infrastructure needs, 

economics, and environmental impact and life cycles analysis would be expected to be addressed in 

follow-on projects once siting selection questions are addressed. 1.2.1. Our study is based on having 

conducted thorough reviews of the relevant literature. We have conducted an extensive literature reviews 

on macroalgae and biofuels both prior to and during this study. These have resulted in two documents: (1) 

Roesijadi et al. 2008. Techno-Economic Feasibility Analysis of Offshore Seaweed Farming for Bioenergy 

and Biobased Products, PNWD-3931 (175 references) and (2) the FY 2010 deliverable, Roesijadi et al. 

2010. Macroalgae as a Biomass Feedstock: A Preliminary Analysis. Technical Report 19944 (73 

references). The former covered biology of seaweeds, the seaweed industry, biobased seaweed products, 

offshore seaweed farms, structures and technologies for offshore farms, environmental factors, 

environmental impacts, seaweed biotechnology, and preliminary techno-economic feasibility. The latter 

added preliminary resource analyses of production in the United States; the macroalgae-to-market supply 

chain (cultivation, harvest, preprocessing, types of biofuels, and conversion technology for macroalgae); 

preliminary economic assessment; and preliminary life cycle analysis. The initial steps for developing the 

GIS model concept involved a separate literature review of algal species and relevant production data, 

infrastructure needs, and competing uses based on both algal operations as well as other marine and 

coastal operations. Spatial analysis approaches in aquaculture and studies on macroalgal production were 

reviewed. This literature forms the basis of our analysis. We conduct ongoing searches routinely. 1.2.2. 

The conceptual macroalgae growth model includes critical variables including solar radiation 

(photosynthetically active radiation), turbidity/light penetration, sea surface winds, water temperature, 

salinity, and nutrients 

1.3. NC 1.4. NC 1.5. NC 1.6. NC 






No progress, even on literature assessment. 


The presenter stated (mistakenly), that no macroalgal production now exists in the US. It is important to 

review the published literature on of current or recent harvesting and aquaculture projects. The project 

should give more attention to realistic amounts of sea surface, biomass to gasoline conversion and amount 

of gasoline needed. Among the critical factors not currently in the model would be the depth, storm 

frequency, types of algae (biomass potential) that could be grown or harvested in different areas of the US 

exclusion zone. 

The model did not appear to include any cost factors for offshore development of algae. 

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Provided preliminary assessment of macroalgae as biofuels feedstock for the United States. Demonstrated 

with selected data and test criteria that data for the bio-physical model and selection constraints for the 

competing use models can be mapped for the west and east coasts of the United States. 




Delivered macroalgae review (preliminary analysis) in FY10; in FY11 goal is to produce specific 

resource assessment tool. Unfortunately, the depth of the analysis is unclear. It would have been helpful 

to have a better overview of historic and international activities (and previous analyses). 



It appears that the growth model needs critical review, refinement and validation prior to application. 


2.1. Our year 1 deliverable was an initial review of the literature and preliminary analysis. For 

development of GIS tools, a literature review was completed and initial approach identified. This is 

consistent with the timeline developed for this project. 2.2.1. There is currently no macroalgae cultivation 

industry in the U.S. for biomass production at the scale needed for biofuels. There is cultivation activity, 

but not at levels to constitute an industry. In the FAO Fisheries and Aquaculture Statistics 2006, the U.S. 

is not listed as a contributor to world production of cultivated macroalgae. With few exceptions, open 

water cultivation of macroalgae in the U.S. has been experimental, demonstration, or pilot-scale. The first 

lease for open water cultivation in the State of Maine was recently granted to grow kelp for a specialty 

market. Likewise, the U.S. contributes only 0.5% of the worldwide harvest of natural stands. The largest 

commercial wild kelp harvester ISP Alginates (formerly Kelco) discontinued its leases of kelp beds in 

California in the 2005-2006 timeframe and ceased operation. 2.2.2. In project year 1 and the FY10 

deliverable, a proof of concept calculation provided an order of magnitude estimate of the area of 

macroalgae cultivation for a nominal gasoline replacement of 1% of the current domestic consumption 

based on best available data. We estimated about 11,000 square kilometers (equivalent to the area of the 

Island of Hawaii) would be needed, illustrating the scale of production needed. We also reviewed 

literature on aquaculture projects, developed a needs matrix, and interviewed industry scientists. Based on 

this, we identified two groups of kelp as a focus for this year’s assessment: Macrocystis and 

Saccharina/Laminaria and related species. Selection was based on species distribution, growth rates, and 

biochemical feedstock derived from the literature and our own research. The purpose of this assessment is 

to provide the tools/models to generate a critical baseline for which to base future research questions. 

Cultivation at the scale above will need to be offshore on suspended scaffolds to avoid competition with 

nearshore kelp populations. Depth is not critical because the species would be suspended near the surface, 

but wave energy associated with storm frequencies is. In the 1970s, the demonstration project of offshore 

Macrocystis production off Southern California saw loss due to late fall storm events. We agree that there 

is a need to address storm event frequency and in particular, wave heights. Storm track frequencies, in 

addition to remotely-sensed wave height data are publicly available to help develop probabilities of 

occurrence that can help assess a prioritization score of candidate sites. This work requires the use of 

foundational building blocks to establish the overall assessment. If at this point there is reasonable 

potential, we can then work to develop and integrate additional factors providing a more rigorous 

assessment. 2.2.3. Our current focus is on biomass production potential and developing a GIS spatial 

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analysis tool to map this potential. This is expected to provide a good baseline to assess potential across 

U.S. waters. Differential costs of development across the landscape can be included later to calculate the 

net value in specified areas. 2.3. NC 2.4. NC 2.5. Our previous analyses, including international and 

historic activities, are documented in our FY10 report “Macroalgae as a Biomass Feedstock: A 

Preliminary Analysis”. 2.6. We agree that the macroalgae growth model needs critical peer-review and 

validation. The current model considers energy balance components including PAR and light and water 

quality aspects such as water temperature, salinity, and turbidity. Growth rate and other biophysical 

parameters for Macrocystis and Saccharina/Laminaria will be derived from peer-reviewed literature. 

Observation data from the literature and field sites will be used for validation. 







The project should be viewed as not relevant without the requisite preliminary analysis. 


Much of the information related to this project already exists in the literature. Its assembly and modelling 

has merit if realistically done. 


There is a need to evaluate macroalgae as biomass resource to enable Biomass Multi-Year Program Plan 

goals. 




The project is relevant to strategic goals. 



Relevance of survey results is contigent on validation of growth and sustainable harvest models. 


3.1. NC 3.2. The macroalgae growth model requires critical peer-review and validation. Observation data 

from the literature and PNNL's own field sites will be used for model validation. 3.3. NC 3.4. NC 3.5. NC 

3.6. We agree that the macroalgae growth model requires critical peer-review and validation. The current 

model considers energy balance components including PAR and light attenuation as well as water quality 

aspects such as water temperature, salinity, and turbidity. Growth rate and other biophysical parameters 

for Macrocystis and Saccharina/Laminaria will be derived from peer-reviewed literature. Observation 

data from the literature and field sites will be used for model validation. Results from the macroalgae 

growth model can be used to guide decisions around harvest intensity 

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assessments) 




end-use) 




The possible deliverables were presented as the CSFs, instead of a real assessment of the CSFs. 


A strong review of the relevant literature combined with realistic costing of offshore macroalgal 

development etc. are critical to success. Note that Chinese macroalgal industry data should not be used (I 

mention this because of FAO data that the PI might find and consider using); labor costs are insignificant 

there, but more importantly, the Chinese shelf is spatially extensive and very shallow (a few meters depth 

for kms of distance from shore), which is one reason why the industry is so successful. 


There may be no successful outcome from this project, please see concerns listed under "Overall 

impressions" 






The criticality of model validation apears to be under-emphasized. 


4.1 NC 4.2.1. Economic analysis will be needed for a global assessment of macroalgae as a feedstock. 

The current project is centered on assessment for macroalgal growth conditions and competing uses. 

4.2.2. Direct comparisons are not a complete crosswalk. This is a new potential feedstock, and, for 

understanding the overall potential, we need to use comparisons and knowledge from a variety of 

industries ranging from algae, other forms of aquaculture, and marine energy to come up with the most 

reasonable assumptions. As to data from the Chinese industry, those production estimates are the basis for 

comparison with production from current experimental kelp cultivation efforts underway in other regions, 

such as in Ireland. Those production estimates compare favorably with those of the Chinese kelp industry 

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and, together, form a basis for comparison for production potential through cultivation. Activities in 

China would not provide a strong basis for economic comparison for the reasons stated by the reviewer. 

4.3. A study by Graham et al.( PNAS 104: 16576–16580 [2007]) demonstrated the existence of 

previously unknown populations of deep water tropical kelp resulting from model predictions based on 

ecophysiological modeling and geospatial analysis of physico-chemico data from ocean environments. 

This is not unlike the approach we are taking. The two demonstration areas are being used to test model 

parameters. Once validated for prediction of population distribution in the demonstration areas, the plan is 

to apply the model to broader aspects of the U.S. EEZ. Our success in developing and applying such 

spatial models is listed as a challenge in our presentation. However, members of our team have 

successfully developed parallel models for land-based resource assessment of microalgae applicable to 

the United States (Wigmosta et al.2011. National Microalgae Biofuel Production Potential and Resource 

Demand. Water Resources Research. doi: 10.1029/2010WR009966). We feel that this experience will 

facilitate research on the current project on macroalgae. 

4.4. NC 4.5. NC 4.6 The macroalgae growth model will provide information on growth rates and total 

biomass. Understanding how much of the resource can exist is needed for its valuation. Perhaps the 

reviewer is noting that other factors are needed as well to assess a realistic long-term production potential. 

We agree that model validation is important. This study is a first step, and the first of its kind to provide 

quantitative macroalgae production potential as a biofuels feedstock. 







I did not sense the coordination with the OBP Algae program or with other investigators. 


Tech transfer not evident yet in presentation. 


Publish findings and disseminate new knowledge to end-users. Provide new assessment tool to end-users. 

Submit results to the Biomass Knowledge Discovery Framework. 




No technology transfer reported. 


See section on Overall Impressions 


Information sharing appears premature. the survey needs validation and refinement before it is useful. 

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5.1. This is one of two projects focused on macroalgae as a marine biomass in the OBP algae program. 

Members of our research team are the lead and participants of the “Microalgae analysis” project in the 

OBP algae program. The other project is the “Collaborative: PNNL -Canada Collaboration”, also being 

conducted at PNNL. The principle investigator of the current project is also on the research team of the 

collaboration with Canadian scientists. 5.2. NC 5.3. NC 5.4. NC 5.5. NC 5.6. NC 5.7 We have just started 

the GIS tool development and assessment. FY10 was concluded with delivery of the report “Macroalgae 

as a Biomass Feedstock: A Preliminary Analysis”. This report was provided to staff of the Bioenergy 

Knowledge Discovery Framework. The GIS development and assessment is, in effect, a separate and 

focused activity that builds on the preliminary analysis. The deliverable for FY11 is a project plan for 

research to be concluded in the following years and preliminary assessment. Data will be shared when 

models have been appropriately applied and reviewed. 


Please provide an overall evaluation of the project, including strengths, weaknesses, the project 

approach, scope, and any other overall comments. 



This project seems highly unrealistic and has not had the basic analysis performed to justify the program. 


Better modelling and better input data relative to macroalgae are critically needed; the current models 

appear to lack many relevant factors. 


This project appears to be advocating cultivation and/or harvesting of algal biomass from the coastal 

regions of the US. If the latter, large tracts of ocean floor would be needed for the biomass-to-biofuels 

conversion. Is the harvesting of the coastal kelp forests the equivalent of clear cutting in forests? Has the 

PI addressed ecological and environmental consequences of such a plan? This may be a good exercise on 

paper but a moot point in practice, as the ecological consequences of large-scale harvesting of algal 

biomass from coastal regions would likely be untenable in the context of viable biofuels. There is a startup 

company in Texas that aims to exploit Macrocystis pyrifera upon cultivation on land, in 

photobioreactors for biofuels generation. This may be an ecologically and environmentally more 

acceptable approach. 



Macroalgae for fuel has been the subject of much research in past decades. The task "Conduct a high level 

review of biomass feedstock and conversion potential of macroalgae for biofuels production in the United 

States" was completed. What were the results of this review? What are the current and projected technical 

and economic feasibilities of macroalgae biofuel production? That should be established before the 

resource assessment. 


Nice presentation of a straightforward project. Given the scarcity of data on macroalgae conversion, some 

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of the underlying assumptions in the model seemed unlikely. It is unclear how helpful this assessment 

will prove to be. 


This was a terribly underassuming talk that did not articulate any compelling argument for the work. It's 

an interesting subject, though, and one would hope the project is better managed if continued. It is simply 

difficult to provide much more than that in commentary based upon what was presented. 


This appears to be an early first step in developing a meaningful macroalgae feedstock resource analysis. 

The incompleteness of the work needs to be emphasized to put the status of the survey in proper 

perspective. 


6.1. NC 6.2 This project has two primary modeling activities that are being developed in parallel: first, the 

macroalgae biophysical growth model and spatial assessment, and second, the competing use and 

infrastructure constraints model. The macroalgae growth model considers energy balance components 

including PAR and light attenuation as well as water quality aspects such as water temperature, salinity, 

and turbidity. Growth rate and other biophysical parameters for Macrocystis and Saccharina/Laminaria 

will be used from past and current lab and field studies conducted by team members who are experts in 

the field, as well as through current peer-reviewed literature. Selection of the kelp was based on growth 

characteristic, known biochemical feedstock constituents, and current international activities that have 

identified the brown macroalgae as suitable marine biomass. Observation data from the literature and 

PNNL's own field sites will be used for model validation. The conceptual framework for the spatial 

assessment has been established such that we can conduct analysis at the 1km spatial resolution for U.S. 

EEZ. While not a part of the formal operating plan at this point in time, additional considerations for 

storm track frequencies and wave heights can be factored in and used to help establish a prioritization of 

areas where macroalgae growth is favorable. Growth model results will be used in conjunction with 

spatial modeling of competing use analysis, regulatory requirements, and existing/required infrastructure 

to further refine the prioritization of potential sites and economic viability. The team consists of 

established experts in marine spatial planning and GIS modeling, mass and energy balance modeling, 

geoinformatics and spatial modeling, marine phycology and algal ecology, ecological risk assessment, 

and marine biotechnology. 6.3. We are not advocating development in coastal regions or harvest of 

natural populations. In fact, we are inclined to follow the recommendations of the Pew Oceans 

Commission that marine aquaculture for the United States should focus on offshore, rather than 

nearshore, waters. Our goal is to provide an unbiased national resource assessment for cultivation of 

macroalgae as marine biomass as a contribution to national biofuels feedstock production goals stipulated 

in the Energy Independence and Security Act (EISA). Environmental aspects including habitat and 

species are being considered under “competing uses” in our assessment. 6.4. NC 6.5. NC 6.6. NC 6.7. We 

discuss the relatively early stage of our GIS model development and application and provide explanatory 

background above. The GIS model development is a new task initiated in FY11. 

Project: 9.6.1.2 PNNL 

Title: Microalgae Analysis 

Presenter: Mark Wigmosta 

Presentation Date: Friday, April 08, 2011 

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Approach 5.43 1.29 7 

Progress 5.57 1.18 7 










The approach is quite reasonable. 


Modelling study based on production of 220 billion L/yr of algal biofuels. Designed to aid in site 

selection for farms. 

The presentation did not clarify what productivity levels (what algal species? what productivity figures?) 

are being input to the model. The PI notes that the water resources' database is from 1978 and presents a 

problem in modelling effort. The basic unit selected for model seems reasonable (100 farm ponds of 10 

acres each plus 200 acres for infrastructure). 

The model should include additional sensitivity analysis in two ways: 1) Ground-truthing of sites selected 

by the model as good is needed to see if the model is really performing, and 2) the model needs to 

incorporate sealevel rise predictions/changes in predicted precipitation patterns on a decadal level for the 

next 30 years, at least. 


A systematic biophysical evaluation of resource demands and constraints on microalgae biofuel 

production. 




The objectives are to continue development of resource demands and constraints on microalgal 

production. The approach and targets are technically sound. 


This is detailed but misses the crucial application of experience of application. The approach is 

exhaustive and pays good attention to detail, but its a bit too sanitized. If the PI's could integrate the view 

from the ground, i.e. the farmer, into their criteria this would become an excellent study. I encourage the 

PI's to include this perspective and turn this work into an excellent product for the DOE. 


The project should be structured such that all major factors affecting the potential production are defined 

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and accounted through the Integrated Assessment framewok before specific numerical results are 

presented and published. 


[1.1] N/C --- [1.2a] Biomass productivity was estimated using a biophysical growth model based on 

incoming solar radiation and water temperature determined from local meteorology through energy 

balance calculations. Biological input parameters to the model were derived from current literature and 

are explicitly described in Wigmosta et al. (2011). As noted the last systematic national water resource 

assessment was conducted in 1978. Part of our current effort is to estimate water availability with the 

most current information at each potential pond site. [1.2b] We agree that ground-truthing is an important 

issue for assessments of resource use and algal biofuel production. We have approached this issue at 

several levels including: 1) In the early phases of our efforts in developing the Biomass Assessment Tool 

(BAT) we worked closely with J.R. Benemann and D.B. Anderson in developing the conceptual design, 

data requirements, and underlying assumptions for the algal growth model which is the foundation for the 

biomass production and resource use estimates produced by the BAT. 2) Simulated pond evaporation 

rates from our model were compared with corrected pan evaporation data over a range of climate zones. 

3) To the extent possible, based on limited published literature values and anecdotal information provided 

by J.R. Benemann derived from his direct experiences in algae cultivation, we have attempted to 

benchmark our analytical results. For example, our results indicate an average conversion efficiency of 

total solar energy to organic mass of approximately 1%. This is consistent with the published value of 

1%-3% for observed yields from Williams and Laurens (2010). We also found our calculated national 

mean annual vegetable oil production rate of 5775 L/ha/yr is conservatively well below the published 

value for current production of 14,000 L/ha/yr by Mascarelli (2009). We have plans, during the summer 

months of 2011 to visit several research sites where algal growth experiments are planned or underway 

including: 1) PNNL's Marine Science Laboratory where current lab experiments represent different U.S. 

regional climates and focus in on both fresh and saline water algal strains; 2) University of Arizona where 

focus is on the design and operation of raceway's in arid climates using freshwater species; 3) Texas 

A&M (Pecos) to evaluate large-scale pond systems and production strains for biomass and lipids; and 4) 

Utah State University where research is focused on cold weather cultivation and alternative nutrient 

addition strategies. We will also monitor algal growth experiments at New Mexico State University and 

Texas A&M (Corpus Christi). [1.2c] Using the GIS capabilities of BAT, it would be relatively 

straightforward to consider the impacts of sea level rise to evaluate the sensitivity of microalgae 

production potential to alternative climate change scenarios, particularly related to land availability in the 

coastal regions. --- [1.3] N/C --- [1.4] N/C --- [1.5] N/C --- [1.6] We agree with the invaluable need to 

ground-truth from the perspective of operators/farmers; please see Response [1.2b]. --- [1.7] We agree 

with the reviewer in that it is ideal to model the major factors in multiple stages of the lifecycle process, 

as is being done under the INL/PNNL Integrated Assessment Framework (IAF) project. There are 

however, clear benefits to the scientific research community to share periodic and logical progressions 

within specific lifecycle stages; the current project has benefited from such presentations and publications 

from others, and our assumptions, results and contributions have been peer-reviewed by experts in the 

field and published in Wigmosta et. al (2011). It should also be noted, that the IAF leverages major 

components developed under the current project allowing a model progression to include and represent 

the "resource to production to refinery" portion of the lifecycle. 


Please evaluate the degree to which the project has 

made progress in its objectives and stated project management plan 

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has met its objectives in achieving milestones and overcoming technical barriers 



The progress is adequate to the time spent on the project so far. 


Progress in this project is satisfactory, but the things mentioned in the approach section would likely 

improve the project. 


Development of a GIS-based Biomass Assessment Tool that integrates high-resolution data with 

biophysical models to provide spatially/temporally explicit estimates of production potential and resource 

demands and constants 




Scientists have not finished different modules, but indicated that milestones will be accomplished by the 

project deadlines. 


No problem - good work progress. 


Progress falls significantly short of the state that should be required before specific estimates are 

presented and published. 


[2.1] N/C --- [2.2] See Response 1.2(a-c) --- [2.3] N/C --- [2.4] N/C --- [2.5] The project in on track to 

meet project milestones and deadlines. --- [2.6] N/C --- [2.7] The research, methodology, and products 

developed under the current project provide key building blocks to realizing the ability to represent the 

full lifecycle process. We feel that sufficient progress has been made to establish national baseline 

conditions under well-specified assumptions and limitations (Wigmosta et. al 2011). As such, there are 

clear benefits to the scientific research community to share periodic and logical progressions within 

specific lifecycle stages; the current project has benefited from such presentations and publications from 

others, and our assumptions, results and contributions have been peer-reviewed by experts in the field and 

published in Wigmosta et. al (2011). 

Wigmosta, M. S., A. M. Coleman, R. J. Skaggs, M. H. Huesemann, and L. J. Lane (2011), National 

microalgae biofuel production potential and resource demand, Water Resour. Res., 47, W00H04, 

doi:10.1029/2010WR009966. 





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This project has high relevance for the OBP program 


This kind of model is very important to DoE efforts to reach affordable, sustainable production of algal 

biofuels in US. 


This study provides DOE and industry with the first spatially-explicit estimate of open pond microalgae 

production potential and resource demands for the conterminous United States. It realistically addresses 

critical questions of how much energy can be produced, where production can occur, and how much land 

and water resource will be required. 




The project is highly relevant to platform goals. However, the ultimate utility of this tool will depend on 

the transparency of its assumptions (and the ability to change them). 


Relevance will shoot up to 9 or 10 once they incorporate the follow-on perspective of the farmer. 


A survey that includes estimates of all major factors has great relevance. The current state of this survey 

has signficant omissions. 


[3.1] N/C --- [3.2] N/C --- [3.3] N/C --- [3.4] N/C --- [3.5] The Biomass Assessment Tool (BAT) webinterface 

allows a user to access, query, and interact with all national datasets and models produced under 

this project. Ultimately, the user will have control to adjust spatial filtering parameters as well as model 

parameters to screen existing databases for elements such as minimum suitable land area, geographic 

boundary, proximity to nutrient source, proximity to CO2 source, climate ranges, biomass production 

rates, lipid production rates, percent of national production target, and temporal ranges. The BAT is 

constructed such that when more current or detailed knowledge is available for particular algal strains in 

specific geographic domains this information can be incorporated. We have intentionally designed the 

BAT and the algal growth model such that we can readily incorporate more accurate information or 

improved models for specific algae strains, environmental conditions, and downstream lifecycle processes 

as they become available. For example, we are closely monitoring ongoing and planned technology 

development efforts by the NAABB Algal Biology and Cultivation Teams where there is a focus on 

increasing overall productivity of biomass accumulation and lipid/hydrocarbon content and increasing 

overall productivity by developing and optimizing scalable cultivation practices and growth rates under 

various environments. We also are interacting with PIs at PNNL, Utah State University, University of 

Arizona, and Texas A&M to evaluate their results, and as appropriate, incorporate them into the BAT. 

Finally, we are continually reviewing the algae and bioenergy literature to identify potential 

enhancements to BAT as appropriate. --- [3.6] We agree with the need to incorporate farmer/operator 

experience. This is part of current and planned future work; please see Response 1.2b. --- [3.7] The 

current state of our study provides a high spatial/temporal evaluation of open pond microalgae production 

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potential along with water and land requirements for feedstock growth when water and nutrients are not 

limiting. Ongoing work (WBS 9.6.1.6) is prioritizing and optimizing sites that are best suited in the 

"resource to feedstock to biorefinery" portion of the lifecycle by considering nutrient supply, CO2, water 

demand and availability, and system logistics including cost of infrastructure. Unique aspects of different 

algal strains will also be considered under NAABB allowing the capability to optimize strains to local site 

conditions, including climate, water source, nutrient requirements, and production in an open/closed 

system. We anticipate that the results of this study will supply valuable data for use in complete life cycle 

analyses. 








assessments) 




end-use) 




The CSFs and Challenges are stated reasonably well and very pertinent. 


Realistic ground-truthing of model predictions (and revision of model, if needed) and incorporation of 

sealevel rise predictions will help to assure that the results are useful. 


Integrated assessment of on-site feedstock production, integration into full lifecycle analysis 




Critical factors have been formally identified, but one factor that appears to be missing is any 

observational component to "ground-truth" some of the results. Without this component, this is likely to 

be an interesting modelling exercise with very limited practical application. 


Very good attention to the resource criteria. Very good. Score will shoot up as soon as the PI's incorporate 

logistic criteria of on-site review. 

Very good assumptions made. This is a very realistic study. I have high confidence in these PI's. 

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Critcal success factors should include development of an accurate Integrated Assessment Framework. 


[4.1] N/C --- [4.2] We agree that ground-truthing is an important issue for assessments of resource use 

and algal biofuel production; this is the subject of current and planned work as detailed in Response 1.2b. 

Using the GIS capabilities of BAT, it would be relatively straightforward to consider the impacts of sea 

level rise in the coastal regions of the national assessment, thereby providing an evaluation of the 

sensitivity of microalgae production potential under alternative climate change scenarios. --- [4.3] The 

research, methodology, and products developed under the current project provide key building blocks to 

realizing the capability of representing the full lifecycle process. The foundation of this realization is 

based upon the scalability of the BAT to link lifecycle components in an integrated and flexible manner. 

Under WBS 9.6.1.6, this capability is being exercised to bring PNNL-developed spatial analysis and 

production models together with the INL-developed system logistics model enabling a "resource to 

production to biorefinery" representation of the lifecycle. --- [4.4] N/C --- [4.5] We agree that groundtruthing 

is an important issue for assessments of resource use, algal biofuel production, and observational 

validation of modeled results; this is the subject of current and planned work as detailed in Response 1.2b. 

--- [4.6a] We agree on the need for logistics criteria. Ongoing work (WBS 9.6.1.6) is prioritizing and 

optimizing sites that are best suited in the resource to feedstock to refinery portion of the lifecycle by 

considering nutrient supply, CO2, water demand and availability, and system logistics including existing 

and required infrastructure. [4.6b] N/C --- [4.7] The research, methodology, and products developed 

under the current project provide key building blocks to realizing the capability of representing the full 

lifecycle process. The foundation of this realization is based upon the scalability of the BAT to link 

lifecycle components in an integrated and flexible manner. Under Project 9.6.1.6, “Integrated Assessment 

Framework”, this capability is being exercised to bring PNNL-developed spatial analysis and production 

models together with the INL-developed system logistics model enabling an integrated "resource to 

production to biorefinery" capability. 







There was proposed interaction with NAABB specifically along with some small dissemination plan for 

the literature. Somehow this should be made more available to the academic and investigative 

communities. 


Presentations of model should continue. This model should be very important to other DoE-funded 

projects. 


Integration of web-based Biomass Assessment Tool into Bioenergy Knowledge Discovery Framework, 

coordination with NAABB, collaboration with INL & ANL 

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Technology transfer partners (within NAABB) have been identified but there was no indication of any 

current interaction with them. The project would be benefit from early and frequent interactions with 

potential users. 


No problems here. 


Tech transfer should have been limited to within the Integrated Assessment Framework team until a 

complete an accurate model is developed. Presentation and publication of survey results prior to 

validation may be counter-productive. 


[5.1] Selected results from NAABB will be integrated into the Biomass Assessment Tool (BAT) and be 

available to the academic and investigative communities (as well as industry and other interested parties) 

though its web-based interface (additional detail is provided in Response 3.5). Results of this project will 

also be disseminated through BAT linkage to the DOE Bioenergy Knowledge Discovery Framework 

(KDF), as well as, peer-reviewed papers and technical reports. --- [5.2] We will continue to disseminate 

project results; please see Response 5.1 for more details. --- [5.3] N/C --- [5.4] N/C --- [5.5] We are 

closely monitoring ongoing and planned technology development efforts by the NAABB Algal Biology 

and Cultivation Teams where there is a focus on increasing overall productivity of biomass accumulation 

and lipid/hydrocarbon content and increasing overall productivity by developing and optimizing scalable 

cultivation practices and growth rates under various environments. We also are interacting with PIs at 

PNNL, Utah State University, U of Arizona, and Texas A&M to evaluate their results, and as appropriate 

incorporate them into the BAT. In addition, results, methods, and models developed under this project 

will be distributed through the BAT and the KDF. --- [5.6] N/C --- [5.7] We feel there are clear benefits to 

the scientific research community to share periodic and logical progressions within specific lifecycle 

stages; the current project has benefited from such presentations and publications by others and our 

results and contributions have been peer-reviewed by experts in the field and published in Wigmosta et. al 

(2011). 






The project has a good feel to it. Information of value to the development of algal biofuels will be an 

outcome. 


The project is proceeding satisfactorily. 

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Are there any conclusions drawn from the work so far? A tasks, milestones and timetable slide would be 

useful to add. Is the analysis dependent on the biological system, i.e., the species to be used? Greenhouse 

covered ponds, which allows controlled energy and water vapor transfer with the atmosphere, would be 

an untenable situation for biofuels production, as it will make the process cost prohibitive. A lot of this 

work has already been done ad infinitum by the "race track" and "photobioreactor" field. Thus, work in 

this project is redundant, as it repeats what is already in the literature. 



CO2 is probably the most limiting resource, but it seems to be near the last that is going to be 

implemented. Similarly, it's not clear how much has been accomplished on the Biomass Growth Model. 

Presumably it would show that the upper tier states cannot produced economical quantities of biomass 

due to a short growing season. That would eliminate the need for the detailed analysis of the other 

resources in these regions. Hopefully, not much effort was expended to model land and water resources in 

non-viable locations (i.e., no CO2, wrong climate). 

The land available would be expanded somewhat by considering degraded cropland rather than excluding 

all cropland. 

It was not clear if the water use considered blowdown disposal in addition to evaporation. 

Ground-truthing of some specific sites should be done early in the program to have an idea of how many 

false positives/negatives are being generated. 

Important project. 


This is a good project, but it would be even better if potential users were consulted frequently and some 

sort of "ground-truthing " of the results could be incorporated. The presenter seems well aware of the 

model's limitations and is willing to improve the model's transparency (with respect to basic 

assumptions). 


A very good start to a very important study. The PI's are encouraged to incorporate the added perspective 

of the farmer and land developer to further down select sites that pass the resource nexus criteria. Will 

become a very important study once this is completed. 


The survey has very high potential value but needs further refinement and validation and integration into 

a broader assessment model before it is useful. Key assumptions need to be challenged and reviewed. 

Uncertainities need to be estimated and included when meanignful quantitative resutls are presented. 

Publishing that 17% of U.S. transporation fuels could be produced by algae should be discouraged when 

it is know that including techno economic factors makes the error range of the estimate 0% to 100%. 


[6.1] N/C --- [6.2] N/C --- [6.3] An important step to realizing the potential of algae is quantifying the 

demands commercial-scale algal bevel production will place on water and land resources. This study has 

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provided baseline estimates of land potentially suitable for open pond microalgae feedstock production 

along with water demand for various levels of biomass production. Initial results suggest that microalgae 

based biofuel has the potential to make a significant contribution to renewable fuel targets, however, 

water demand is likely to be high. Final results are ultimately species dependent which is represented in 

the growth model through input parameters that control the conversion of solar energy to biomass/lipids, 

in particular those associated with water temperature and light intensity. Impact of CO2 and other 

nutrients is the subject of current work in WBS 9.6.1.6. We chose a greenhouse configuration to represent 

the general features of closed systems in terms of temperature and evaporation control. The main focus of 

this study is a first step to identify resource demands and constraints on the production of microalgae, not 

the engineering details of open or closed systems. Our study presents a first of its kind high‐resolution 

spatiotemporal national assessment that brings to bear fundamental questions of where production can 

occur, how many land and water resources are required, and how much biofuel could be produced. 

--- [6.4a] Regarding CO2, see Response 3.7. We agree that growing season length needs to be considered 

and is currently being implemented under WBS 9.6.1.6. The biomass growth model provides hourly 

estimate of biomass production that will allow us to quantify biophysically and economically viable 

growing seasons. [6.4b] Our GIS-based land screening analysis is flexible and will allow inclusion of 

additional rulesets, including degraded agricultural land using metrics available in the USDA-NASS 

database. [6.4c] Our current study of saline water includes water requirements for blowdown; 

consideration of blowdown disposal is proposed for FY12. [6.4d] Re. the need for ground-truth data – see 

Response 1.2b. [6.4e] N/C --- [6.5] Re. the need for ground-truth data – see Response 1.2b. --- [6.6a] We 

agree with the need for ground truth data and operator perspective – see Response 1.2b. [6.6b] N/C [6.6c] 

See Response 1.2a for biomass productivity. Also, we are currently refining the model to account for CO2 

and other required nutrients. [6.6d] N/C [6.6e] N/C [6.6f] Water use was calculated from our open pond 

model and is roughly equal to the water required to replace evaporative loss less precipitation. Biofuel 

production was based on biomass calculated from our growth model with a lipid content of 20% and an 

extraction/conversion efficiency of 80%. [6.6g] We are currently developing an estimate of available 

water supply that will be compared to site-specific freshwater demand. [6.6h] Our enhanced growth 

model will allow consideration of atmospheric CO2. [6.6i] N/C [6.6j] See Response 6.3. --- [6.7] We 

agree that the BAT needs further refinement to consider additional resource constraints and additional 

components of the lifecycle (see Responses 3.7, 4.3, 4.6a, 4.7, and 6.4). We agree this project will benefit 

from ground truth data and additional input from algae farmers/operators (see Response 1.2b) as part of 

the model review, refinement, and validation. We also agree with the need to estimate uncertainty; this is 

a subject of proposed research for FY12. As noted in Response 1.7, we do feel there are clear benefits to 

the scientific research community to share periodic and logical progressions within specific lifecycle 

stages; the current project has benefited from such presentations and publications by others, and our 

assumptions, results and contributions have been peer-reviewed by experts in the field and also 

contributed back to community in Wigmosta et. al (2011). 

Project: 9.1.3.1 INL 

Title: Algae-Based Biofuels Integrated Assessment Framework: Development, Evaluation, and 

Demonstration 

Presenter: Deborah Newby 



Approach 4.43 1.05 7 

Progress 4.57 1.29 7 


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The Approach builds upon previous feedstock supply modeling work which is a good starting point. 

Development of a model which allows for the cost productivity analyses to be performed for various 

operating schemes is desirable. One aspect that I may have mistaken is hardwiring in certain processing 

steps which are known to be too expensive. I hope that is not the case. 


This modelling project will construct an Algae Logistics Model (ALM) of an Integrated Assessment 

Framework (IAF). BAT is included. These model components will be put together, and require lots of 

careful project coordination between partners. 

The model used flocculation and centrifugation in the model, simply because modelling data were 

available; however, these processes may make the process of recovery too expensive at the scale required 

for real algal biofuels' production. PIs need to mine older literature (e.g., Oswald, earlier DoE reports) for 

data to make a realistic (and productive) model. 

The species of algae that are cultured will make a big difference to models; I suggest PIs try to model at 

least one specific model. PI had concern that relevant "proprietary data" is making modeling difficult. Try 

to avoid this problem by seeing if current DoE-funded projects' can share data plus use older, published 

literature. 

The scale of the pond that is modelled may be too small. 


Asystematic biophysical evaluation would be provided of resource demands and constraints on 

microalgae biofuel production 




The goals of the project are to develop an algae logistics model and to provide baseline designs for algal 

systems. There seems to be considerable overlap with the NREL project "Algal biofuel pathway baseline 

costs." 


Models can be a waste of time if the system modeled is nonsensical. The PI has followed the Solix image 

of a large microalgae farm in the middle of the desert surrounding a pilot plant. The obvious problem is 

that such a plant will not likely be built, as power plants are not generally built in the middle of nowehere. 

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Is this near a coal supply...etc. And what about distribution lines for the electricity generated? What is the 

energy balance once all that has been factored in...etc. 

I would suggest that the PI's go back to the reports of Benemann et al, and Lundquist et al. Model those 

systems and then expand. 


Much of this assesment appears to be redundant with NREL Algal Biofuel Baseline Costs and other 

published cost and resource analyses. 


The approach builds upon previous feedstock supply modeling work conducted at the INL and consists of 

essentially two main components: 1) development of an algal logistics module (ALM) that will leverage 

the history of supply system modeling at the INL; and 2) integration of this expanded feedstock supply 

model with the co-funded work being conducted at PNNL expanding their Biomass Assessment Tool 

(BAT) to include algae. The completed product will be the Integrated Assessment Framework that will 

allow us to select appropriate locations for algal farms based on specific criteria including land value, 

slope of land, sunlight, temperature, water availability, etc. and then to model the logistics and costs 

associated with the placement of a particular farm design at that site. Co-location or proximity to other 

resources such as waste-water treatment plants or existent coal-fired plants as a source of CO2 as well as 

proximity to transportation infrastructure and processing facilities are will also be considered by the IAF. 

There is no expectation of building plants adjacent to algal farms, but through the use of the IAF, sites 

with appropriate characteristics will be identified for siting of algal farms. The Solix model was not the 

basis for our design, which as the reviewer suggests is not economically viable. We agree that it would 

make sense to site algal-farms near water resources such as the Gulf Coast or wastewater treatment plants. 

We also agree that modeling existing algal farms is essential both from design and validation 

perspectives. We are working to incorporate such designs into our model databases. The algae logistics 

module consists of numerous data layers that can be tailored to a particular site and configuration. No 

aspect of the design is hardwired, allowing the model to be dynamic as new engineering solutions to the 

challenges in algal biofuel production are developed This logistics modeling effort does provide cost data 

for system design concepts in a similar fashion to previous techno-economic analysis efforts including the 

assessment performed by NREL. The most important products of this work are not economic analyses, 

but rather the establishment of an engineering decision making framework that can couple suites of 

models and data sets supporting the development of viable algae supply system technologies and designs. 

Economic evaluations of system performance are clearly a critical component of establishing viability and 

are one of the important assessment metrics delivered through the modeling toolkit. As noted by the 

reviewers, achieving cost and performance targets required to make algal biofuels a reality will require 

engineering advancements to existing technologies and systems. The modeling framework being built 

through this work is strictly focused on providing the ability to determine performance at the engineering 

fidelity required to support new technology and concept development. 





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The Project has only been operational for a short time. 


The partners are making progress but need to evaluate their data sources carefully and improve them, and 

the processes and scale used. 


Development of a GIS-based Biomass Assessment Tool that integrates high-resolution data with 

biophysical models to provide spatially/temporally explicit estimates of production potential and resource 

demands and constants 




Progress is limited because the project began in the fall of 2010. 


In terms of effort and work done, this is fine. 


Preliminary model uses several technologies that have been shown to lack viablity based on technoeconomic 

analysis, and modeling results appear to confirm previous work. 


The modeling framework consists of numerous model and data layers that can be configured to represent 

a particular site and farm design. This allows the model to be dynamic as new engineering solutions to the 

challenges in algal biofuel production are identified. We established the baseline design presented as that, 

simply a baseline. We pointed out that we recognized that the scale of 1-acre farms included in the 

baseline has been shown to not be economically feasible. It did provide use with a starting point to begin 

populating our databases with equipment and energy use specs, and was also used to help us put together 

the linkages between the different elements of the farm, and also for the development of the hooks 

between the different components of the joint model for integration of the logistics analysis (ALM-INL) 

with the biomass resource assessment (BAT-PNNL). The initial phase is essential to the development of 

the model, and raceways and farms of appropriate scale and configurations will be introduced into the 

framework developed through this baseline effort. We agree that the baseline design lacks viability and 

that the modeling effort confirms previous work. Additional farm designs will be incorporated as the 

model is refined, and will yield an integrated assessment that extends beyond currently available models. 

The IAF will provide DOE with a decision support tool for robust exploration of open pond microalgae 

production potential and resource demands for the conterminous United States. 





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Models that help perform technoeconomic reviews without costly experimentation are valuable. Who will 

hve access to the model? Who wll be served by its use? These questions impact the relevance. Also, the 

model has to fit in its framework. In this case, the model must fit feedstock supply and transportation 

scenarios at both ends to be valuable. This may help define the entire technology package to the refinery 

gate, which would be very relevant. 


The project needs to demonstrate that its models are realistic (see Approach). 


This study provides DOE and industry with the first spatially-explicit estimate of open pond microalgae 

production potential and resource demands for the conterminous United States. It realistically addresses 

critical questions of how much energy can be produced, where production can occur, and how much land 

and water resource will be required. 




The project is highly relevant to program goals. 


The modeling effort is fine, in terms of competence of work, but the process the PI selects should be more 

realistic. Instead of the model of a power plant based farm in the middle of desert-nowwhere one should 

focus on the pond systems in the Gulf coast states or sewage treatment plants. 


The integrated assesment has great potential value. The production and harvesting model built around 

available data from non-viable operations compromised the relevance of the work. It is not clear how 

much of the framework developed will be applicable to other scenarios. Acceptance of the modeling and 

sharing of data and concepts from other institutions are necessary for relevance. 


We agree with the reviewer’s comment that the Solix model is not relevant (see response to Approach 

section). We are building a framework to explore a vast range of design concepts. Establishing this 

framework requires setting a baseline design case to lay the foundation for the model and data integration 

infrastructure. We appreciate and absolutely acknowledge the reviewers’ opinions that the baseline design 

case has performance flaws. The relevance of the baseline design scale of 1-acre may be applicable to 

inoculation pond assessments, but there are clear challenges using this scale as a replicated unit 

throughout a production farm. The basis for design concepts incorporated into the framework will be 

existent farms, as information is available, as well conceptual designs developed from the multiple 

integrated database layers within the dynamic model. Relationships are being developed to gain access to 

information from those operating both commercial and test-bed algal farms. With the completion of the 

modeling framework, investigation of a comprehensive set of design concepts will be facilitated, 

including the ability to seamlessly incorporate technologies being built across the OBP research platform 

and broader research community. The IAF meets many of the DOE roadmap goals for modeling 

including the ability to combine disparate databases, optimize facility location and supply chain logistics, 

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include a number of approaches, scales, and levels of detail required for a particular assessment, and to be 

easily modified to incorporate new technologies as they become available. In terms of access and use of 

the modeling framework, the primary use will be facilitated through an IAF web-based interface that will 

build upon the current BAT web tool housed at PNNL. The IAF interface will include dynamic access 

and utilization of the logistics modeling framework. The logistics toolset will also be available in a 

standalone version at the end of the project. The exact configuration of the user interfaces is not yet 

determined, as the focus has been delivering the logistics modeling capability with the IAF. 








assessments) 




end-use) 




These appear to be adequately defined. 


The project needs to have outreach to some of the other, larger DoE projects' data to make sure model 

variables are realistic and scaled; part of this concern can be met by better reference and use of existing 

data in the literature. 


Integrated assessment of on-site feedstock production, integration into full lifecycle analysis 




Critical factors have been identified, but it is not clear that the project can overcome the challenge of 

gaining access to (and publishing) engineering systems data from successful companies and developers. 


The PI has identified perhaps too many process unit operations at each critical process step (harvesting, 

dewatering, extraction...) with no apparent understanding of how these operate or fit together. I did not 

see the critical energy criteria identified in the algal biofuels roadmap. The PI would do well to read this 

and the reports of Benneman et al and Lundquist et al to the DOE to get a much better framework for 

critical criteria. 

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critical success factors are well defined. It's all about partnership and sharing to determine viablity by 

integrating process concepts and locations. 


As noted by both presenter and reviewer, a key critical success factor is the ability to access designs and 

data from established algal farms. The research team is working to develop relationships with others 

working in the area of algal biofuels in order to gain access to engineering systems data from successful 

companies, developers, and other research institutions (see response in section 5). The Algae Road Map 

notes that access to proprietary information associated with algae pilot and production facilities is a key 

challenge. The PI’s appreciate the reviewer’s concern about the complex interactions between process 

unit operations. Rigorous mass and energy balance constraints are being implemented for every process 

element within the supply system modeling framework. The PI’s will refer to the reports as suggested for 

further reference and cross check on critical criteria. 







The model should link in very well with the overall process of delivering bioproducts from algae, 

providing the correct modeling hooks are included, and it is flexible enough in construction to allow for 

modifications. 


The potential of the project for useful tech transfer wouldimprove a lot with better model input data. 


Integration of web-based Biomass Assessment Tool into Bioenergy Knowledge Discovery Framework, 

coordination with NAABB, collaboration with INL & ANL. 




Technology transfer will be a major challenge for this project. 


Limited but this a PhD project. 


Specific plans to partner with institutions developing process technology are needed. 

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The PI’s appreciate the comments about providing the correct modeling hooks, and having flexible 

construction. The PI’s are deeply engaged in communities developing advanced data and model 

integration tools for engineering decision making, and are building the algae logistics modeling 

framework with current state of the art concepts and tools. In fact, the tools and approaches being utilized 

in this modeling framework have been associated with three R&D 100 awards over the past five years. 

We agree that technology transfer will be a challenge for this project and the progress and 

accomplishments to date have not been focused on technology transfer. This is because the most 

important knowledge and conclusions that will be developed by this project are not yet known. The PI’s 

are very aware of the role and need for technology transfer, and will be continuously working with project 

partners and DOE to make sure the right pathways for technology transfer have been identified and are 

being pursued. The research team has begun working to develop relationships to gain access to 

engineering systems data from successful companies, developers, and other research institutions. Current 

partnerships with entities operating large-scale ponds or farms include those with Utah State University, 

Energy Dynamics Laboratory, and Pacific Northwest National Laboratory and their NAABB partners that 

operate ponds at University of Arizona and Texas A&M. This will give us access to a variety of scales 

ranging from 220L- 24,000L raceways, and a floating pond configuration at 154,310 L. Algal include 

strains of Chlorella, Nanochloropsis, Chaetoceros, Neochloris, and indigenous/wild fresh water strains 

from Utah. We are working to expand our relationships with commercial algal farms as well and 

appreciate the suggestion offered by one reviewer that Cyanotech Corporation would be willing to 

provide information. 






This modeling work seems to promise a useful product when completed. One hopes all available 

commercial data is utilized in the model, and that some plan for testing the full scale model is part of the 

end deliverable. 


The project may need to use additional data and scales. 


Are there overlaps between Biomass Assessment (PNNL) and Algae Logistics Model (INL)? Does the 

analysis depend on the species cultivated and the product derived? E.g., the species used would determine 

Algae harvesting processes. PI needs to bring this important aspect into the modeling. The alga biotech 

sector has already accomplished and currently use these criteria on a routine basis: -Raceway 

infrastructure -Pond configurations -Product storage and transportation are issues that have been solved. 

How is this theoretical analysis going to be better than what already has been accomplished by the private 

sector? Reviewer recommends that the PI should go to the Big Island of Hawaii (Kona) and visit 

Cyanotech Corporation. They would be happy to help you get a head start. Overall, this appears to be 

redundant work. It is reinventing the wheel. 


Overall Impressions: 3 

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Goals listed in presentation are numerous and presented is somewhat disjointed fashion: Technology gap 

analysis, cost analysis (aka techno-economic analysis), resources analysis, infrastructure compatibility, 

algae logistics module (similar to woody biomass analysis), and baseline facility designs, 

Harvesting and dewatering technologies selected thus far are mostly conventional and obviously too 

expensive and/or energy intensive for algae biofuel. Pond sizes being assessed are 1 acres and 10 acre. 1 

acre is obviously too small to be economical. 

I don't see how this technology analysis and model designs are different from the many others in the 

recent literature and in Aquatic Species Program reports. Isn't this also somewhat a duplication of the 

technology assessment being done by NAABB/NREL? 


The apparent overlap with an NREL project was disconcerting (at the very least, they should collaborate). 

The challenge in gaining access to proprietary engineering information seems very problematic and there 

was no clear path to overcoming this obstacle. 


Overall this project needs more realism in terms of the systems it models. 


The challenge of acquiring scenarios supported by pilot data appears to be the greatest hurdle. The 

integrated assesment framework appears to be a valuable, or perhaps essential tool for production 

scenario evaluation and selection, but it is only useful if validated process data is available. building the 

tool around available data instead of relevant data does not appear to help advance the technology. 


Many of the overall impression comments have been addressed (see respective sections above). When 

completed the IAF will provide DOE with the first spatially-explicit estimate of open pond microalgae 

production potential and resource demands for the conterminous United States. It will meet many of the 

DOE roadmap goals for modeling including the ability to combine disparate databases, optimize facility 

location and supply chain logistics, include a number of approaches, scales, and levels of detail required 

for a particular assessment, and to be easily modified to incorporate new technologies as they become 

available. Data from pilot scale and commercial algal production farms will be incorporated based on 

availability of data through literature, partnerships, and collaborations as described previously. As 

suggested by the reviewer, a full testing and verification plan will be delivered as part of the conclusion to 

the initial two year project. These activities will be performed to the degree that resources allow through 

the development process, and a comprehensive testing matrix will be established for potential continued 

work. We have read and continue to refer to the foundational literature indicated by the reviewers. These 

papers reflect the state of technology applications at the time the reports were generated. Our model is 

designed to encompass those technologies (and clearly it should be consistent with them) but also to 

expand beyond those technologies; and combine spatial resource information with production and supply 

chain logistics. INL and NREL have begun collaborating in this area and look to leverage unique aspects 

of each modeling approach. The modeling framework being developed at the INL in support of the IAF is 

providing a plug-and-play architecture for investigating a broad range of technologies and design 

concepts for algae supply systems. This approach is a natural extension of several initial techno-economic 

assessments that have been performed to date. Furthermore, the logistics modeling framework being 

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developed in this work provides an opportunity for exploring engineering research questions using model 

and data tools at multiple fidelities as may be required by the specific question at hand. 


Title: Collaborative: Algae-based Integrated Assessment Framework 

Presenter: Richard Skaggs 



Approach 5.57 1.18 7 

Progress 5.86 0.99 7 










The Approach in this project is well reasoned. 


This is a modelling project that incorporates features such as the cost of land. Productivity is based upon 

the canonical Redfield ratio, but it is now well known that different algal taxonomic groups (e.g. green 

algae vs diatoms) have different Redfield ratios. It would be good to try to do a general model, and then 

test it against a specific case, and adjust Redfield ratios appropriately for a species that is an actual 

biofuels' target (e.g., Nitzschia as a diatom; Chlorella as a green alga). 


Effort provides integration and systematic biophysical evaluation of resource demands and constraints on 

microalgae biofuel production with harvesting logistics and cost analysis coupled with sound project 

management. 




The approach is to build CO2 and nutrient source data bases for inclusion into a DOE analysis 

framework. The approach is sound and feasible. 


The approach appears reasonable but its very difficult to differentiate if from the previous PNNL 

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presentation, and that from INL (Newby). It seems to duplicate that work, and its difficult to see the 

interconnection. 

That said, its an important resource model and these efforts are important. However, a much more defined 

relationship to the other PNNL model would be recommended. 


The project management plan would benefit from some in-progress validation process, perhaps including 

virtual and real visits to sites deemed suitable for alagla biofuel production. 

Definition of goals for model accuracies might have value. 


1.1 N/C 1.2 The canonical Redfield ratio is a valuable starting point for incorporating nutrient use into our 

general model. As with any model, keeping the underlying assumptions in mind is necessary for 

interpreting the model. It will be beneficial to ensure the model is built with flexibility in regarding later 

adjustments in the C:N:P ratio. After completing the general model, adjusting the C:N:P ratio to reflect, in 

turn, a diatom and a green alga may be good next steps. The caveat to these iterations of the model would 

be that different strains are likely to be employed in different climates and latitudes, so we will utilize 

different strain-specific ratios as appropriate and as data become available. Furtrher, future algal 

biological and cultivation research may provide data and understanding leading to a more mechanistic, 

improved approach to modeling nutrient use. We are collaborating with the NAABB Algal biology and 

Algal Cultivation Teams as they generate new information on algal productivity as a function of nutrient 

sources and use and climatic conditions. 1.3 N/C; 1.4 N/C; 1.5 N/C; 1.6 Related projects are WBS 

.9.6.1.2, “Microalgae Analysis,” and WBS 9.1.3.1, “Algae-Based Biofuels Integrated Assessment 

Framework: Development, Evaluation, and Demonstration,”. WBS 9.6.1.2 is responsible for much of the 

original Biomass Assessment Tool (BAT) development, as well as ongoing selected enhancements 

including analysis of freshwater availability, saline water sources, web-based interface development and 

linkage to the KDF. WBS 9.1.3.1 is, by design, a collaborative project with WBS 9.6.1.6 such that the 

BAT’s GIS-based, high-resolution resource assessment capabilities are coupled with INL’s Algae 

Logistics Module (ALM) to create the Integrated Assessment Framework (IAF). In short, INL’s efforts 

are focused on development and delivery of the ALM, including providing pioneer supply system design 

for feedstock logistics encompassing algal harvesting, storage and transport, and cost and engineering 

performance assessment. The IAF will provide a resource assessment capability that addresses the 

complete feedstock supply chain. The 3 tasks within WBS 9.6.1.6 are distinct in scope from the 2 projects 

above. Task 1, being executed by PNNL building on the existing BAT, is focused on addition of critical 

data base and analytical elements to the BAT. Task 2, being executed jointly with INL, is to develop the 

IAF, coupling the high spatial and temporal modeling capabilities of BAT with ALM. The result will be 

an analysis framework encompassing the complete microalgae feedstock supply chain. Task 3, working 

with INL, is using the IAF to conduct selected national, regional, and site-specific analyses of microalgae 

production potential and tradeoffs. 1.7 Validation is an important issue for resource use and algal biofuel 

production assessments. For this project and the overall BAT development, we have approached this at 

several levels: 1) Early on we worked with J. Benemann and D. Anderson to create the conceptual design, 

data requirements, and underlying assumptions for the algal growth model 2) Based on limited published 

values and anecdotal information provided by J. Benemann, we have benchmarked our analytical results. 

For example, our results indicate an average conversion efficiency of total solar energy to organic mass of 

approximately 1%. This is consistent with the published value of 1%-3% for observed yields from 

Williams and Laurens (2010). 3) During 2011 the team will visit research sites where algal growth 

experiments are underway: a) PNNL lab experiments representing different U.S. regional climates for 

fresh and saline water strains; b) U. of Arizona focused on design and operation of raceway's in arid 

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climates using freshwater algae; c) Texas A&M to evaluate large-scale ponds and production strains; and 

d) Utah State University focused on cold weather cultivation and nutrient addition strategies. 







Progress is acceptable for the length of time thus far. 


The models need to incorporate the newer data being produced (try to get input factors from several of the 

larger DoE projects) and use information from the published literature, including on Redfield ratios. 


Progress has been achieved on growth model and data bases development, land value model, and IAF 

framework design. 




Progress is limited because the project just started. They have accessed EPA databases for powerplants 

and wastewater treatment facilities; have started looking at NPDES permit sites and ag-based nutrient 

datasets. 


Good progress and work well done. 


Initial progress is good, but the scope of the model is quite daunting. 


2.1 N/C 2.2 We designed the BAT and the algal growth model such that we can readily incorporate more 

accurate information or improved models for specific algae strains, environmental conditions, and 

downstream lifecycle processes as they become available. For example, we are closely monitoring 

ongoing and planned technology development efforts by the NAABB Algal Biology and Cultivation 

Teams where there is a focus on increasing overall productivity of biomass accumulation and 

lipid/hydrocarbon content. The Cultivation Team is focused on increasing overall productivity by 

developing optimizing scalable cultivation practices and growth rates under various environments. We 

also are interacting with PIs at PNNL, Utah State University, U of Arizona, and Texas A&M to evaluate 

their results and as appropriate incorporate them into the BAT. Finally, we are continually reviewing the 

algae and bioenergy literature to identify potential upgrades to BAT as appropriate. 2.3 N/C 2.4 N/C 2.5 

The nationally available data from EPA and others are being incorporated into the IAF to better define 1) 

additional components in the lifecycle process, and 2) potential co-location/co-benefit opportunities to use 

other lifecycle waste products in the algae process stream. Spatial modeling of these entities, along with 

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required transport pathways and energy requirements, will provide necessary metrics for site 

prioritization. 2.7 The project team agrees with the reviewer's comment. However, in recognition of the 

DOE OBP’s challenge of establishing a credible, accurate inventory of national feedstock resource 

potential and assessing environmentally sustainable feedstock potential, our goal is to develop a feedstock 

supply assessment framework that can be readily updated and improved as new data, models and 

information become available. Thus, over the long-term, the expectation is that many researchers and 

institutions will contribute to addressing OBP’s challenge, and the IAF will provide a framework for 

organizing and utilizing research results, and contributing to the needed assessments. 







This project is extremely relevant to the OBP program on algal biofuels. 


These models will be helpful to establishment of biofuels' farms, but need careful data input and 

sensitivity analysis of models. 


Effort further provides the DOE with analysis framework to holistically evaluate U.S. microalgae 

production potential and associated resource requirements and feedstock logistics 




The project is relevant to platform goals. 


Again, the relevance is predicated on a much clearer integration or collaborative relationship with the 

other PNNL study (and the Idaho model). 


Work is hihgly relevant because an accurate assessment to define alagal biofuel potential is necessary. 


3.1 N/C 3.2 We agree with the reviewer's comment. Please see Response 2.2. We further agree that 

sensitivity studies will be an important tool to understanding the various tradeoffs involved in the siting 

and scaling of industrial-scale feedstock supply enterprises, and the associated resource and infrastructure 

requirements. These sensitivity studies will be conducted under the IAF testing and demonstration efforts 

for this project scheduled for FY12. 3.3 N/C 3.4 N/C 3.5 N/C 3.6 We agree with the reviewer's comment. 

This project is by design a collaborative effort that is integrating PNNL’s BAT with INL’s ALM, and 

utilizes the existing capabilities at both laboratories as a point of departure. In addition, please see 

Response 1.6. 3.7 The overall objective of this project in collaboration with the INL project is to develop 

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an integrated analytical framework to conduct an accurate assessment of algal biofuel potential 

encompassing the "resource to production to biorefinery" portion of the feedstock lifecycle. 








assessments) 




end-use) 




The CSFs and Challenges are adequately explained. 


This model effort needs additional input data. One factor not yet included that is important is the effect of 

sealevel rise and predicted climate change, in general. This should be incorporated as a sensitivity 

analysis. 


Integration of on-site feedstock production and harvesting logistics assessments into full lifecycle analysis 




Critical success factors were identified but it will be a major challenge to make the model truly useful. 

How to develop a model with enough depth in the different modules so that it is practical (and not just an 

exercise in modeling using GIS overlays)? 


I saw no problems here. 


The scope of the model is so vast that critical success factors include focusing on key leverage items 

where there are gaps in current knowledge and in the selection and incorporation of best available exisitng 

data and models. For example, it appears that the processing component of the logistic model could be 

improved and moved forward by using NREL modeling. 

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4.1 N/C 4.2 See Responses 2.2 and 3.2 regarding quality of input data. As to potential impacts of climate 

change in general and sea level rise in particular, these are not part of the current work scope of this 

project, however, ongoing work at PNNL (WBS 6.2.1.2 Global Change Assessment Model and WBS 

11.1.1.6 Biomass Production Under Climate Change) is evaluating the potential impacts of climate 

change on water availability on overall biomass production, and specifically water availability. Regarding 

sea level rise, for future efforts it would be relatively straightforward using the GIS capabilities of BAT, it 

would be relatively straightforward to consider the impacts of sea level rise in the coastal regions of the 

national assessment, thereby providing an evaluation of the sensitivity of microalgae production potential 

under alternative climate change scenarios. 4.3 N/C 4.4 N/C 4.5 The models and analysis capabilities 

housed under the BAT are designed for flexibility and usability in terms of adding additional/updated data 

sets and linking additional models, such as is being done under the current project. This is accomplished 

in the BAT design for two fundamental modes of operation. First, the 'workstation' version of the BAT 

enables full access to the data, models, configuration, and parameter files allowing a researcher to adjust 

and tune the various models and data at will. Second, the web-based BAT is intended to provide a userfriendly 

interface to interact with, analyze, and configure and execute the included biophysical and spatial 

models. Our intent with the web-based BAT is to get beyond the complexities of the 'workstation' BAT 

and allow a tool that is easily used and readily available for private industry and others. The project team 

consists of several Geographic Information Science professionals bringing to bear state-of-the-science 

spatial models and analysis. Several ongoing projects are supporting the BAT development process to 

achieve the goal of having a single analysis portal that is benefited by the work of the many ongoing 

collaborations. 4.6 N/C 4.7 N/C 







There is good coordination for the project and within the project. 


This project has potential for good technical transfer. 


Web-based Integration Assessment Framework into BioenergyKnowledge Discovery Framework and 

collaboration with INLand NAABB 




Technology transfer partners have been identified but it is too early to evaluate progress. 


Same comment on interaction with other PNNL study. That is needed to improve tech transfer value. 

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Strategic partnersip with industry and NAABB should be developed to the fullest extent possible. 

Premature publication of estimates should be avoided. Any estimates that are eventually reported should 

include error estimates and uncertainties. 


5.1 N/C 5.2 We agree with the reviewers’ comment. An explicit element of our approach is to make the 

IAF both accessible and generally useable through a web-based user interface and by direct linkage to the 

KDF. Also see Response 4.5. 5.3 N/C 5.4 N/C 5.5 See Response 5.7 below. 5.6 N/C 5.7 The project team 

agrees with the reviewers’ comments. Team members are directly involved with the NAABB and 

associated industrial partners. We are diligently identifying areas for synergy, collaboration, and 

data/information/model exchange. 






This project has value for the Program as a whole. 


This modelling approach needs to be more realistic to be useful, but has the potential to be an excellent 

effort. 


Are there overlaps between the two Biomass Assessment efforts at PNNL? and also with the Algae 

Logistics Model effort at INL? Such analyses (including the previous two presentations) are premature 

given the uncertainties that surround strains, products, processes, all of which do not enter into the 

"Assessment". It is better to invest the funds in the biology of "Algae Biomass", where our collective 

knowledge and expertise are severely lacking. The concern here is that by the time a microalgae process 

can be scaled up, "models and associated spatial and temporal information for improved understanding of 

geographic location, relative cost, and sustainability of potential microalgae biofuel production" may be 

obsolete. 


Overall Impressions: 6 

This project is another adder to the GIS-ALM project. This project adds the crucial CO2 and nutrient 

resource data, with the CO2 being the most limiting resource that probably should have been the starting 

point of the three projects. Why are these three separate projects, each with their own budget? 

Skaggs: 2 years $700k 

Newby: 2 years $700k 

Wigmosta: 1 years, $365k 

The Redfield Ratio does not apply to algae cultivation, only to observations of marine algae. How does 

this project (and the Wigmosta project) build on the Sandia resource assessment? For this and the Newby 

project, what references are going to be the basis for your facility designs and resource uses? The PIs 

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should consult extensively with experts and not rely on the highly variable quality of published literature. 

As with the Newby project, I suggest some ground-truth designs be prepared early in the project. 


Good project but its practicality is likely to be limited by the depth of its individual modules. 


See above. 


The assesssment is valuable but challenging. Modelling must have flexibility to incorporate rapidly 

changing technology. Key leverage items should be indentified early in the modelling effort so resources 

can be focused. Model validation strategy needs to be developed. premature dsitriburtion and use of 

modelling or resutls is a significant risk. 


6.1 N/C 6.2 See Response 1.7. 6.3 As described in Response 1.6, there are three principle tasks within 

WBS 9.6.1.6 that are distinct in scope from the other projects. Task 1, is adding critical data base and 

analytical elements to the BAT, namely: ▪ CO2 and nutrient source data bases ▪ Enhanced algal growth 

model to estimate nutrient and CO2 requirements, and ▪ Land value and availability model and data base 

Task 2, coupling the BAT with ALM consists of two primary elements: 1) develop the IAF design 

specifications and enabling architecture requirements, and 2) integrate the software and data base 

components of the BAT and ALM, develop necessary new components (e.g., proximity/cost model, 

feedstock supply tradeoff model, etc.), and web-based IAF user interface. Task 3, joint (w/INL) 

application and demonstration of the IAF will evaluate at regional/national scales: optimal 

feedstock/harvest enterprise locations and scales infrastructure requirements and identification 

regional/national production performance and feasibility 6.4a WBS 9.6.1.2, “Microalgae Analysis is an 

ongoing effort, including the initial development of the BAT that originated in FY 2008. This project and 

WBS 9.1.3.1 are new projects initiated in September FY10 with distinct scopes of work to build on and 

extend the BAT capabilities as described in Response 1.6 and 6.3. Thus, DOE OBP has funded them as 

separate projects. 6.4b See Response 1.2. 6.4c The teams for this project and WBS 9.6.1.2 have consulted 

with with Ron Pate from the early stages in the development of the BAT to ensure this project builds on, 

without replicating the work of Ron Pate and others at Sandia National Laboratory. As part of his March 

17, 2011 presentation to the NRC Study on Algae Biofuels Sustainability, Ron provided a comprehensive 

side-by-side comparison of the Sandia and PNNL algae biofuels resource analyses. For example, he 

characterized the PNNL assessment as “high resolution/granularity at National level w/ 30-m resolution” 

based on “approximate physics-based growth modeling based on conditions” and the SNL assessment as 

low resolution/granularity at State level, multi-state regions” and “no detailed physics-based modeling.” 

6.4d As discussed in Response 1.7, we are making every effort to ground our work based on first hand, 

real work experience in microalgae production with a particular emphasis on current and planned 

laboratory and pilot scale efforts. 6.5 We agree with the reviewers’ comments. Therefore, we have 

designed the BAT in general such that we can readily incorporate more accurate information or improved 

models for specific algae strains, environmental conditions, and downstream lifecycle processes as they 

become available. It is also our expectation that by designing the IAF to enable broad accessibility, over 

the long-term many researchers and institutions will contribute to the depth and breadth of the individual 

IAF modules. 6.6a See Responses 1.6 and 6.3. 6.6b See Response 6.4c. 6.7 See Response 1.7. 

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Project: 9.6.1.1 National Renewable Energy Lab 

Title: US-Israel Algal Biofuels (NREL) 

Presenter: Robert Baldwin 



Approach 5.29 1.67 7 

Progress 5.14 1.55 7 










The project is basically about evealuating whether microwaves can be used in algal conversion to some 

advantage. I would prefer the approach be focused. The attempts to deal with the 

development/demonstration/commercial problems are premature in my view. 


This project aims to demonstrate cost-effective extraction of lipids from algae, and has tried pyrolysis, 

novel extraction methods, gasification etc. The feedstock types being evaluated are Nannochloropsis and 

Coccomyxa, with microreactors to scale models. 


Interesting and potentially useful approach at extracting oils from algal biomass via pyrolysis. 




The project's goal is to characterize a specific set of extraction and conversion methods in collorabation 

with a commercial source in Israel (Seambiotic). Methods include microwave extraction, gasification, and 

pyrolysis. It seems appropriate to characterize these methods, but this seems like an academic exercise 

(i.e. no breakthroughs expected). 


Microwave assisted extraction is a challenge for large scale commercial extraction. 


A preliminary techno economic analysis may have eliminated certain process options such as microwave 

heating, allowing resources to be focues on potentially viable options. 

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The objective of the project is to evaluate methods other than the traditionally-used solvent extraction to 

see if they can be applied to selectively release desired components from algal biomass. The original 

experimental plan was to proceed with a few targeted experiments, then down-select based on technical 

results. This plan was followed for all 3 technologies being researched: 1) solvent extraction - 

hydrothermal; 2) pyrolysis; 3) gasification. A considerable amount of data has been collected for all three 

strategies but could not be adequately presented in the less than 10 minute timeframe that the presenter 

was allowed. 

A persistent criticism of the work is that it is not supported by technoeconomic analysis (TEA). We wish 

to point out that the research was exploratory in nature and was designed to develop information (e.g. 

process conditions, product yields, etc.) that can now be used to develop a TEA. Our primary goal was to 

compare several low-severity processes based on technical merit, and eliminate from consideration 

processes that were found to be technically infeasible (such as gasification due to high tar loadings); the 

project is only now at a maturity level where valid economics can start to be developed. To label as 'bad 

science' exploratory research that may be later found to be economically challenged is unfair. On the 

other hand, to use pre-conceived notions of economics to do process downselect before any data has been 

collected is simply misguided. 







Progress is perhaps behind schedule, based on the extent presented. 


The project began in March 2009, and has completed work that shows that nitrogen contamination of 

some lipids is a problem requiring solution in their extraction techniques. 


Some promising results were presented for both hydrothermal and pyrolysis; gas cleaning for syngas 

product may be problematic. 




The project appears to be on target with its stated goals. 


Not a lot of data - appears to be one-off experiments. 


Progress might have been compromised by lack of concept screening using preliminary techno economic 

analysis. 

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If the project were focused to deliver conclusive information on the pluses and minuses to using 

microwaves as source of power or directed energy input, this would be highly relevant. The longer it 

takes to come to conclusions, the less relevant he overall project is. 


Efficient extraction of lipids from good feedstock is very important. 


A mixture of long and short-chain hydrocarbons that would be extracted by pyrolysis might fit the need 

for renewable biofuels from algal biomass. This effort supports DOE objectives for HC transportation 

fuels from biomass 




The project is relevant to platform goals. 


There was an apparent lack of hypothesis driven work that justified the choices made. 


There appears to be a high level of uncertainty if some the process options are viable even if the bench 

scale chemistry is successful. 









assessments) 

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end-use) 




Critical success factors were not enumerated. It sounded like investigators were looking for them. I think 

they have not evaluated what are the real key deliverables, and what it would take to achieve them. 


Procedures need to be refined to produce "clean" lipids. 


Success factors: Low severity processing has opportunity to provide low cost pathway for extraction of 

high value fuels precursors Challenges: separations and gas cleaning 




Among the critical success factors is "cost effective methods for production of fuel precursors." 

Overcoming this obstacle seems unlikely (even at the outset) using these technologies. 


From a scale up perspective, this approach is extremely challenged from an energy balance or process 

control point of view. 


A critical success factor is techno economic screencing of process concepts prior to initiation of lab work. 

This was not evident in the presentation. 


A criticism was leveled concerning the economic viability of using microwaves. We agree that if 

microwaves are used simply to heat the water in the algae/water slurry that this is an inefficient and 

(likely) uneconomical prospect. The reviewers failed to grasp that our hypothesis is that microwaves can 

be used selectively to effect cell wall rupture by hydrothermal degradation, thus liberating lipids which 

will spontaneously separate from the water phase and spent algal bodies (solids). Further, we have found 

that the aqueous phase contains most of the protein material from the cell walls. Thus, the microwaveassisted 

process is a way of effecting simultaneous extraction and separation of lipid materials from algae 

- this is the hypothesis being tested. This is being accomplished at very low severity conditions compared 

to other hydrothermal processes which operate at pressures of about 150 atm and temperatures around 

350°C. Our work on pyrolysis is similarly providing evidence that fuels precursors can be effectively 

liberated from algae under low severity conditions. Gasification was found to produce a syngas with high 

tar loadings, so this pathway has been de-emphasized in the research program. 

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The project is a collaboration, and coordination seemed to be adequate. The results need to be shared with 

the other OBP programs. 


There is a high potential for effective tech transfer from this project. 






Technology transfer potential is very good because of the commercial collaboration. 


I did not see much in this regard. 


The presentaition lacks evidence of colaberation in indentifying viable opportunities for thermochemcial 

options. 







I believe with focus, this project would have real value. This is true no matter the outcome. If welldesigned, 

even negative results allow decisions to be made in the overall process. 


The chemical extraction procedures being devised and tested are important and the project personnel are 

making good progress. The feedstock targets being analyzed are appropriate. 


Interesting and worthwhile approach. I recommend support and continuation, as pyrolysis appears to 

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provide promising results. Could you elaborate on the conditions employed in pyrolysis? (How do you 

disperse the biomass? Is it wet or dry? Under ambient or other pressure?) 



Of course, economics and environmental considerations are the mostly likely "show-stoppers" and much 

should be known about these considerations based on past research and experience. In this project, early 

efforts should be made to evaluate the prospects for safe economical processes and then track the 

development progress against the needed results. Are the needed levels of improvement conceivable? 

Novel extraction methods are mentioned several times but no details are given. 


The project is technically sound but it was very unclear why the investigators expected to end up with a 

commercially viable technology. 


I note a lack of energy balances in the future work. 


The project appears to have moved to laboratory work before a rigorous screening of concepts was 

completed. The risk of laboratory success delivering dead on arrival technology because of techno 

economic considerations appears to be very high. 


In response to Reviewer 3, we have considerable micro-scale data on dried material. We will perform 

some gram-scale tests in order to obtain enough liquid product to do a more thorough analysis. Benchscale 

fluid-bed pyrolysis with pneumatic feeding will be performed if resources allow. 

Reviewer 6 appears to be concerned about the energy balance of microwave extraction. We are 

performing a series of tests to compare the energy-efficiency of various liquid extraction procedures 

including microwave extraction. 

Project: 9.6.5.1 Sandia National Labs 

Title: Pond to Wheels Algae Biodiesel Life Cycle Assessment 

Presenter: Howard Passell 



Approach 4.71 1.16 7 

Progress 4.57 1.29 7 



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The approach is described. One might argue with some of the choices, but there is at least reasonable 

disclosure. 


The goal of the project is life cycle assessment for Dunaliella production, with evaluation also of 

Nannochloris and Nannochloropsis. Some model parameters come from GREET with cultivation and 

harvesting from Seambiotic. The consumption of electricity for algal cultivation and harvesting is 

demonstrated by the model to be highest impact cost and by their assumptions a 100,000 m2 facility is 

optimal. The model needs to include other factors (e.g., construction costs, provision of power plant). 


•Pond-to-wheels life cycle assessment of CO2, N2O and CH4 emissions for light and heavy duty vehicles 

across cultivation, harvesting, extraction, separation, conversion and combustion. •Provide a model that 

can be shared with other institutions. 




Goals included development of an LCA and a pond flow optimization model using information from a 

commercial process. The approach is sound. 


A solid approach that looks at appropriate process indicators (including energy). It is clear the approach 

was solid as their analysis led to the conclusion that improvements should be reached in 

extraction/dewatering/harvesting. Most LCA are too complicated to relevant to the majority but this 

presentation seemed to be just contained enough that the majority could follow it. I think this approach, 

particularly applied to open ponds has great merit. I also appreciated the fact the LCA was set out outdoor 

ponds which is the likely path forward. 


It appears that the LCA was performed without first completing a thorough review and analysis of 

published LCA's and without selection of relevant scenarios. 

The presentation did not put the pond flow optimization work in context of previous art. 




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The project is closing in on completion. 


Work is 80% complete. Size of optimal facility should be reconsidered in time remaining with additional 

costs added in. 


Milestones completed: Physics model development Physics model validation 




According to the presenter, targeted milestones will be met by the end of FY11. 


Good progress made. 


This project does not advance the state of LCA analysis. 

The presentation did not elucidate advances were in pond modeling. 








It is my view that any number of LCAs and cases and sensitivities are of value to the program. 


These algae are good biofuels' targets so specific models related to data from scaling up facilities fits 

Roadmap. However, failure to consider all costs will limit relevance if not corrected. 


Provides analytical structure for understanding how to reduce energy requirements while increasing 

productivity in algae cultivation. Provides potential improvement to LCA and sustainability indicators, 

and so assists in overcoming barriers identified in LCA 

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Project goals are relevant to platform goals but were limited by the scales considered. 


I found the application of a competent LCA applied to open raceway ponds that incorporate results with 

real energy balances (among other outputs) to be quite relevant. 


Previously published LCA's for algae biofuels are more relevant and complete. 

Pond modeling work should be compared to current state-of-art to illustrate advancemetns being made. 

Specific opportunites for modeling to improve pond design for biofuel scale facilites should be defined. 









assessments) 




end-use) 




Delineation is adequate. One might question whether some alternative choices might have been selected. 


Costs of production may still be too high, even though not all costs are included. If so, consider different 

scales of pond culture. 


Model provides a structure for Seambiotic pond optimization. Seambiotic Model can be useful for all 

algae producers 




Critical success factors were not included in the presentation. 

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Appropriate success factors selected. 


A critical success factor is both pond modelling and LCA is to define the current state of the art and 

ensure that new research advances the technology towards algla biofuel production at scale. 








I believe this result should be a point of discussion with other projects and should also make it into the 

literature. 


Presentations etc. appearing. Model may be useful to other projects if costing factors can be better 

incorporated. 


Plan is acceptable. 






Quite good. 


It is not clear if pond modeling is an advance, and it is not evident how widely the model will be shared. 





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The project is useful. It could have perhaps been improved by a review of some of the assumptions and 

boundaries selected. Nonetheless, it is a valuable contribution. 


This model shows the challenges still ahead for economic production of algal biofuels. Project should try 

to incorporate full costs and do better sensitivity analysis on size of pond and productivity of algae. 


Much of this analysis is available in the photobioreactors literature. 


LCA Overall Impression: 2 

Hydrodynamic Model Overall Impression: 6 

How is this LCA project being coordinated with the Argonne LCA? This project seems to focus on the 

existing Seabiotic plant, which is 1000x smaller than any feasible biofuel plant. In addition, 1000-m2 

individual ponds are modeled, which are about 40x smaller than the size generally considered feasible for 

biofuel feedstock production. The "Future Projection" system is better, but still too small. Use of 

centrifugation for biofuel production is inappropriate on its face. The base-case should be abandoned as 

irrelevant, and the Future Projection should be greatly adjusted. Collaboration with the Argonne LCA 

project may be advisable, not to say that they are using appropriate assumptions either. Any feasible algae 

biofuel process will use only the most simple, low-energy, and largest-scale processes conceivable, as 

outlined in the Aquatic Species Program reports. 

The hydrodynamic-biological model may be valuable for design details on very large raceway ponds. It 

will be good to validate with the small-scale Seambiotic data or other >1000 m2 ponds. 


This seemed to be a reasonable (but predictable) project. It was not clear that this project represents a 

significant advance compared to existing LCAs. 


Quite favorable for the reasons identified above. 


Previous LCA work appears to be more thoroguh and relevant. It is not clear what advances in pond flow 

modeling have been made compared to previous work, and how those modeling advances can improve 

pond design. 

Project: 9.1.2.1 Brookhaven National Laboratory 

Title: New technology: Improving Microalgal Oil Production Based on Quantitative Analysis of 

Metabolism 

Presenter: Jorg Schwender 



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Approach 4.43 2.13 7 

Progress 4.86 1.46 7 










The first step lacking in the approach is a discussion of why this organism is chosen for these detailed 

studies. After all, if this is not likely to be a viable lipid-generating organism, why do the work. 

Investigators suggest it is not in their slide titled Critical Succes Factors. 


This project's goal is to determine how to maximize TAG production from Chlamydomonas. The 

approach includes metabolic studies (e.g. evaluation of acetate enrichment of basic medium vs N 

deprivation) and transformation to add a glucose transporter. 


To address carbon allocation and partitioning in green algae we use: Biochemical analysis Modeling of 

metabolism Metabolic engineering (genetic transformation 




The goal is to identify metabolic targets for increasing lipid production using a model strain of Chlamy. 

The goal and approach seem to have very limited relevance to platform goals. 


This approach (MFA) is far more applicable to the fermentation industry that has the ability to grow pure 

cultures under highly controlled conditions. To apply the MFA analysis of a lab rat strain as a means to 

suggest pond conditions is highly challenged if one agrees open ponds of local cultures is the likely path 

forward. If the work was applied to strains isolated from real ponds, and if the model applied conditions 

that exist in ponds, it might be able to explain why certain productivities are achieved and thus be more 

applicable. 


It is difficult to evaluate if the approach of using Chalmydomonas in acetate media will apply to 

phototrophic growth of other species. 

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All reviewers except reviewer 3 clearly put strongly in doubt the choice of Chlamydomonas as the 

organism to be used in the project. Yet, we believe we can clearly document the relevance of the project. 

First of all, in the algal biofuels roadmap 

(http://www1.eere.energy.gov/biomass/pdfs/algal_biofuels_roadmap.pdf), p. 10, under “Selecting Algal 

Model Systems for Study”, it is outlined that “Species with sequenced genomes and transgenic 

capabilities are the most amenable to investigating cellular processes since the basic tools are in place.”. 

Further relevant document citations see below. In the following the decisive arguments to use 

Chlamydomonas that also have been given in the presentation. We decided to use Chlamydomonas 

reinhardtii (strain cw15) since for this strain we have available well established techniques, mutants, 

knowledge and preliminary results that make the pursued goals attainable: 1) Efficient transformation of 

green algae is not trivial and transformation protocols for Chlamydomonas are already in practice in C. 

Xu’s lab. 2) Chlamydomonas was also chosen because a highly detailed metabolic models have been 

published for this organism (see slide 14, Boyle and Morgan (2009) BMC Systems Biology 3:4; 

Manichaikul et al., Nature Methods 6, 589 - 592 (2009)). A validated metabolic network is basis for flux 

analysis. Although the genomes of other green alga have been sequenced, producing a good metabolic 

network requires a very well annotated genome and would take a considerable amount of time. In 

addition to Chlamy there appear to be genomes of Chlorella sp. NC64A, Chlorella vulgaris C-169 

(Coccomyxa sp. C-169), Volvox carteri, and two marine Micromonas strains available. Yet we could not 

identify any one of these strains to be known as a production strain. 3) The proposal is based on 

preliminary experimental results, such as the accumulation of oil under continuous growth and high 

concentration of the carbon source (acetate). We expect that these findings are transferable to other green 

alga such as Chlorella, Dunaliella or Scenedesmus, but we don’t know for sure until we experimentally 

verify this. We also base the proposal on a starchless Chlamydomonas mutant, which has been 

characterized before in detail by other groups. 4) In the lab of C. Xu intensive Chlamydomonas mutant 

screen efforts are under way that might contribute to our effort. 5) please consider also Morowvata MH, 

Rasoul-Aminia S, Ghasemi Y (2010) Chlamydomonas as a “new” organism for biodiesel production. 

Bioresource Technology 101: 2059-2062 Siaut et al., BMC Biotechnol. 11:7(2011). PMID: 21255402 At 

least we expect that the results and methods generated in these studies with Chlamydomonas will have a 

transformational effect on our ability to optimize oil accumulation in commercial production strains. 

Reviewer 5, claiming that our goals do not identify with the goals of the EERE Algae-to-biofuels 

platform: we disagree. See Algal biofuels roadmap, p. 11, paragraph “Carbon Partitioning and 

Metabolism”. On p. 6 under “OVERCOMING BARRIERS TO ALGAL BIOFUELS: TECHNOLOGY 

GOALS”, the document states as a goal: “Investigate genetics and biochemical pathways for production 

of fuel precursors” See also p. 10 the paragraph under “Selecting Algal Model Systems for Study” Also 

see Multi-Year Program Plan (MYPP): 

http://www1.eere.energy.gov/biomass/pdfs/biomass_mypp_november2010.pdf Taken from p. 2-10/2-11 

of this document, on slide 3 we state that we address the barrier: Ft-C. Feedstock Genetics and 

Development: The productivity and robustness of algae and other feedstocks used for biofuel production 

could be improved by selection, screening, breeding and/or genetic engineering. This will require 

extensive ecological, genetic, and biochemical information, which is currently lacking for most algal 

species. The barrier description mentions the requirement of biochemical information on algae 

productivity. 





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Given the duration of efforts, I would have hoped for more progress. 


The project began recently (October 2010); consequently, few data are available. 


PI utilizes quantitative analysis of flux prior to applying directed manipulation of metabolism 




Limited progress because project began in October, 2010. 


Not a lot of data but the project may have just started. 


Progress needs to be defined agaisnt some specific goals. 


Reviewer 7: “Progress needs to be defined against some specific goals”. We did define specific goals on 

slide 11. See e.g.: 1.1 Establish growth conditions to perform 13C-MFA 90% 1.2 Set up Chlamydomonas 

metabolic model. 90% 1.3 13C-MFA: experimental design (substrate label) 50% 







The relevance is open to question in the investigators own analysis. The slide in the presentation labelled 

Critical Success Factors is not that, but it is a depressing assessment of the relevance and approach. 


The PI suggested that Chlamydomonas might not be the best biofuels' target but that use of it in this 

project would allow techniques to be developed that could then be applied to other algae. I agree that this 

is an excellent model system, but it might be helpful to expand the studies to include a species that can 

both be transformed and is a good biofuels' target. 


•Since the goal is to increase oil yield, research improves the economics of biofuel production (feedstock 

supply). •High oil yields achieved under continuous growth allow for a continuous production process, 

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i.e. simplify the process of feedstock production. •The tools that would be developed (MFA, MCA of 

microalgae) can be further used for directed strain improvement. 




There was no compelling explanation of how the results of this research would be relevant to DOE goals. 


MFA applied to a lab strain and identifying factors (for lipid biosynthesis) will not be particularly helpful 

to open pond production. 


Project should strive to demonstrate early evidence of relevance 


Reviewer 1: The reviewer was asked here to comment about project relevance: Slide 17 discusses the 

relevance of the project. Instead reviewer 1 refers here to slide 18 “4 - critical success factors” and calls it 

“depressing”. We assumed that slide 18 basically was intended to answer the question: What are possible 

reasons why the project could fail? What can be done in each case? For example we describe what could 

be done if Chlamydomonas was successfully engineered but turns out not to be suitable for mass 

production. Reviewer 5: “No compelling argument”. We disagree. The anticipated results are relevant to 

DOE goals. On slide 17 we state: “Since the goal is to increase oil yield, our research improves the 

economics of biofuel production” (feedstock supply). Feedstock supply relates to the MYPP document, 

FT-C. In similar, the algal biofuels roadmap states on p. 6 under “OVERCOMING BARRIERS TO 

ALGAL BIOFUELS: TECHNOLOGY GOALS” as a goal: “Investigate genetics and biochemical 

pathways for production of fuel precursors”. On slide 17 we state: 2) High oil yields achieved under 

continuous growth allows for a continuous production process, i.e. simplify the process of feedstock 

production. Furthermore, the tools we develop (MFA, MCA of microalgae) can be further used for 

directed strain improvement of commercial strains. Related to this, p. 6 of the roadmap document states 

under “OVERCOMING BARRIERS TO ALGAL BIOFUELS: TECHNOLOGY GOALS”: “Achieve 

robust and stable cultures at a commercial scale” Reviewer 6: Our goals are relevant for the algae-tobiofuels 

program. That our results cannot be applied to open ponds is an opinion of the reviewer. 








assessments) 




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end-use) 




These were not enumerated. 


Demonstration of the application of the work proposed to practical algal biofuels' production is important. 

In the future, please report oil concentrations of algae on the basis of algal dry mass, as part of 

presentations (again, important for effective tech transfer) and consider expanding studies to other algae. 

The transformation studies on Chlamydomonas may not be easily transferable to other organisms. 


PI predicts that 13C-Metabolic Flux Analysis might not work well with the two-carbon source acetate. To 

allow Chlamydomonas to grow on glucose, he will transform it with a glucose transporter. While analysis 

is performed with algae growing on acetate, the results of the analysis are to be applied to algae growing 

autotrophically (on atmospheric CO2). Manipulation of the targets revealed by analysis therefore might 

be less relevant for autotrophically grown alga. This issue can be theoretically explored by the use of Flux 

Balance Analysis. Chlamydomonas might not be the most suitable species for mass oil production. A 

succesful engineering approach might have to be applied to another algal strain like Chlorella or 

Nannochloropsis. 




This project is unlikely to have a near-term significant impact since Chlamy is not a relevant organism. 


"If we are successful, Chlamydomonas might not be the most suitable species for mass oil production." 

How is success defined by identifying a strain as not being the most suitable - ignoring the fact this is not 

an industrial strain anyway. 


Applying learnings to potential commerical systems is critical. 


Reviewer1: We disagree. We enumerated critical success factors on slide 18. Reviewer 5: “Chlamy is not 

a relevant organism”: We disagree. See Algal biofuels roadmap, p. 10, under “Selecting Algal Model 

Systems for Study”. This paragraph makes clear that algal model species are not out of the question. 

Reviewer 6: asks “How is success defined by identifying a strain as not being the most suitable - ignoring 

the fact this is not an industrial strain anyway”. We disagree. Again, the algal biofuels roadmap does not 

exclude the use of model species. It says e.g. on p. 10 “Species with sequenced genomes and transgenic 

capabilities are the most amenable to investigating cellular processes since the basic tools are in place.” – 

this clearly refers to our project. 

please consider also Morowvata MH, Rasoul-Aminia S, Ghasemi Y (2010) Chlamydomonas as a “new” 

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organism for biodiesel production. Bioresource Technology 101: 2059-2062 Siaut et al., BMC 

Biotechnol. 11:7(2011). PMID: 21255402 







I see little evidence that this is an integrated project, nor did it seem this was a large concern for the 

investigator. 


The project is in its early stages. 


Transformation of non-Chlamydomonas species 




Potential for technology transfer to producers is extremely unlikely. 


No comment. 


Plan for tech transfer needs to be developed. 


Reviewer 1: We ask DOE management to disregard this very unspecific comment that the PI “seemed not 

to be concerned” about project integration. 

With regards to what was asked of the reviewers to comment about: our project was about 1/2 year old 

when it was reviewed by this panel. There was no chance to get any collaborations or co-publications 

going within that time. 






I appreciate the science that is incorporated in the project. However, this should be work focused on the 

problem of optimized algal biofuel production. The investigators appear to want to do their project 

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whether relevant or not. How this work can then be tied to full scale production has not been developed in 

the plans. 


The project is at an early stage. 


Overall, this is a worthwhile project that should be pursued. Quantitative analysis needs to include 

measurement of the oil content on a per cell or dry biomass basis. Also, information is needed on how 

much oil accumulates in relation to other cellular parameters? Specifically starch and chlorophyll. Slide 

13: TAG/lipid content? How is lipid defined in the context of this work? 



While the technical approach seems reasonable, the use of Chlamydomonas was not defended well. Other 

algae more likely to be suitable for biofuel production have also been sequenced. Why could the studies 

not be done on them? Similarly, the use of acetate and the plan to modify Chlamy to use glucose may 

have scientific value, but the case for eventual value to autotrophic cultivation was not made. 


Although improved basic knowledge of lipid metabolism in algal strains is important, the impact of this 

particular project is unnecessarily limited by the focus on Chlamydomonas. 


The PI has a skill in MFA and an affinity for a given microalgal strain. Thereafter the PI has cranked out 

a classic MFA analysis that does not really relate to the conditions of apparrent absence of what will 

dominate commercial production. 


A clear path from fundemental resutls in a model organism to relevant commercial technology needs to 

be defined. 


Overall: Hard to explain is that none of the reviewer really assessed or criticized the control analysis 

approach (Metabolic Control Analysis, MCA). Yet this was shown in the presentation as a large part of 

the proposal. This effort is complementary to MFA. 

Reviewer 1: “Not relevant for full scale production”: The Algal biofuels roadmap has a wide spectrum of 

barriers and goals defined (p.6). Our project fits “Investigate genetics and biochemical pathways for 

production of fuel precursors”. We see the potential of transformational discoveries that allows to 

overcome the long known principle limitation, that algae typically produce high amounts of lipid 

exclusively under nutrient stress. We have a good chance to address a major problem that already has 

been identified in the Aquatic Species Program. R2: see comments above. R3: In our experiments we 

generally analyze algal cell material by determining major biomass compounds (protein, lipid, starch, cell 

wall material, free metabolites) on a dry weight bases. This may not have been explicit in the 

presentation, but is a basic part of MFA. 

R4: see remarks under “1 Appraoch”. R5: The idea is to gain insights about the role of central metabolism 

in carbon partitioning with the model organism Chlamydomonas, which allows to transfer the resulting 

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insights subsequently to other green algal species. We see the potential of transformational discoveries 

that allows to overcome the long known principle limitation, that algae typically produce high amounts of 

lipid exclusively under nutrient stress. 

R6: The reviewer expresses here his negative view on the value of using model organisms. Also a 

negative view on Metabolic Flux Analysis (MFA), a method usually applied under well controlled 

laboratory conditions. 

Our proposal has a focus on the role of carbon partitioning, i.e. central metabolism in oil accumulation. 

Please note that in the EERE algal biofuels roadmap the goal to understand carbon partitioning was 

emphasized in a paragraph on p. 11. 

We disagree with the view that knowledge gained by MFA with the model species Chlamydomonas is 

irrelevant for commercial production in ponds. Calvin and coworkers dissected the reaction scheme of 

carbon fixation in photosynthesis using green algae under strictly controlled laboratory conditions. Their 

findings are the foundation to understand photosynthesis and biomass production in general as well as for 

green alga in ponds. In addition to the classical Calvin cycle, some years ago the presenting PI could 

describe an important novel functional mode in carbon fixation and oil synthesis (Schwender et al., 

Nature 432: 779–782, 2004). This work was done by the use of MFA and underscores the value of this 

approach for improvement of oil production. 

Please see citations under “1 Approach”. 

Project: 9.1.3.2 INL 

Title: Microalgae Harvesting/Dewatering and Drying 

Presenter: Deborah Newby 



Approach 4.29 0.70 7 

Progress 5.00 0.76 7 










The program approach is reasonable with one big question mark. Did anyone perform a simple energy 

and cost analysis for this particular filtration technology? The technology must have a chance of 

competing with other technologies. The presentation left me with the perception that the technique was 

chosen for other reasons, and no simple assessment was done. 


The project aims to lower the cost of harvesting and collecting algae and to recycle the water used by 

biomass facilities with new/improved processes. Several problems are apparent, but the project only 

began in October 2010, so there is opportunity to improve the project. One problem is that the filter 

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development described is unlikely to fit the scale of pond/raceways required for economical biofuels' 

development. Secondly, the PI is approaching the filtration problem as if unialgal and axenic cultures 

would be filtered; this is unlikely to be true, so "messier" cultures should be used in the work than pure 

Chaetoceros, Cyclotella, Chlorella etc. The PI and team have several scales of raceways built into project, 

which is good, and have previous experiences with filtration problems (nuclear waste to beet processing). 

The algal biofuels' projects have their own distinctive filtration problems, and the PIs need to give 

attention to background knowledge acquisition. 


The work aims to use high face velocity filtration to effect harvesting of microalgae from a culture and 

water recycling/conservation 




The project goal is to characterize and improve on dewatering processes. The specific 

approach is to develop and test different filtration (membrane) technologies. The approach is sound but 

appears limited to one technology. 


This is a technical assessment of a particular type of harvesting dewatering unit operation. In that context 

the approach is fine EXCEPT for lack of energy balances. The PI should present the power consumed per 

unit volume of water consumed from the manufacturer. The PI could have then estimated a typical 

starting concentration of algae in water harvested (about 0.3 gdw l-1) and a typical bio-oil content (about 

20%). From those numbers the PI could point out, right from the start, if this device has a chance to meet 

the target energy load of no more than 10% of the energy (per unit volume processed) as per the Algal 

Biofuels Roadmap. 


A clear definition of project scope and goals in terms of input stream characteristics, output stream 

characteristics, energy consumption, yield, scale, and capital cost target is needed. The goal of cost 

reduction compared to centrifugation is not sufficient. Dewatering economics must enable commercial 

scale algal biofuel production. 


We agree with the reviewer’s comment that the ultimate goal of this technology is to provide a costeffective 

method for dewatering. The selection of the cross-flow technology for application to the 

dewatering issue was based on past work in this area where success was achieved from starting materials 

of similar characteristics to the algae cultures. The presenters also wish to point out that engineering 

solutions, such as development and application of embedded membranes, can significantly improve the 

efficiency and feasibility of using cross-flow membrane technology. This project was funded to look at 

the possibility making these advancements, and we agree an energy calculation is valuable and ultimately 

necessary while noting that the laboratory scale units being used for the development efforts do not 

necessary represent the potential for the technology at scale.. Please see the overall impressions section 

for an energy calculation. As the reviewer points out, the initial parametric testing is being done using 

unialgal and axenic cultures. The basis for this approach is to understand how the different types of algae 

interact with the membrane, so that appropriate membranes may be selected and/or developed for further 

analyses. The initial cultures include 3 green algae, and 2 diatoms, allowing us to assess the membranes 

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with different biochemical and physical properties, including, size, shape, EPS, silica framework, etc. All 

strains selected are known for their lipid production potential. We fully agree the technology needs to be 

assessed using “messier” cultures since that is the reality of what will be harvested from paddle-wheel 

grown outdoor pond algae. We included the variety of scales of raceways in our proposal for this very 

reason. As we develop our membrane dewatering technology our intention has always been to apply it to 

more complex populations. We are currently working with several entities operating large-scale ponds or 

farms include those with Utah State University, Energy Dynamics Laboratory, and Pacific Northwest 

National Laboratory and their NAABB partners that operate ponds at University of Arizona and Texas 

A&M. This will give us access to a variety of scales ranging from 220L- 24,000L raceways, and a 

floating pond configuration at 154,310 L. Algal include strains of Chlorella, Nanochloropsis, 

Chaetoceros, Neochloris, and indigenous/wild fresh water lipid producing strains from Utah. Biomass 

from the variety of ponds grown at Utah State University and the Energy Dynamics Lab, will serve as our 

feedstocks for testing at larger-scale on “messier” cultures. Our hope is that the technology will prove 

useful and we will be able to apply it for dewatering at other production facilities as well. 







It appears a reasonable amount of progress has been made for the project duration. 


This project began very recently. 


Scale and diversity of algal cultures was expanded GC-FID and Nile Red lipid analysis methods were 

applied to expanded algae library Procurement of STC - delivered and initial testing 




Progress is limited since it just began in October 2010. 



Identification of novel technology and intitiation of testing is good progress. Progress on developing 

specific commercial goals and clearly defined harvesting or dewatering concepts is lagging. 


We agree with the reviewer’s comments regarding progress on this new start project. The development of 

specific commercial goals will be addressed as we develop and refine the technology. 

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This project has high relevance if there is likelihood that filtration will be included in the best, most 

effective, lowest cost process scheme. 


Harvesting technologies are critical to success of algal biofuels' projects, but the practicality of the current 

approach will need to be demonstrated. 


Effort aims to improve commercial-scale capabilities in the harvesting and dewatering on microalgal 

biomass 




Project goals are relevant to the Biomass Program Plan but the results may be limited by the choice of 

influent used. Will the results be relevant to paddle-wheel grown outdoor pond algae? 


Unless a quality energy balance is done properly up front this work has little relevance. 


Technological breakthroughs in harvesting and dewatering are needed for algal biofuels to be viable. 

Technologies must be evaluated against commercial scale cost and performance requirements to be 

relevant. 


Please refer the overall impressions section for the energy analysis and to the approach section for 

clarification on strain selection and relevance to populations harvested from outdoor ponds. 








assessments) 



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end-use) 




CSFs have been provided. However, some of them should have been considered to some extent before 

starting the project. For example, to question (as part of the CSFs!) whether there are other economic 

(read more economic) approaches, is a fine example of the analysis that should have been checked before 

start. 


One of the most critical factors to success is starting with a realistic feedstock culture for filtration. The 

project needs to collaborate with biologists who can provide better insight to how these problems will 

affect the goals of the filtration research. 


Critical success factors: Technical – can high shear filtration be performed at an industrial scale for algae 

(1,000’s of liters per hour)? Market – If high shear filtration can be performed is it economic for 

dewatering of algae? Are there other economically feasible drying approaches? Business – Are there 

commercial filtration processes that are economic (with minor modification) for algae and, if so, at what 

$$ per volume? Top technical challenges: Reduction/elimination of membrane fouling (High Shear, 

embedded membranes, and surface charge of membranes) Capture of “phantom” oils (secondary nanofiltration); 

Cost reduction when compared to competition (centrifugation) 




Critical success factors were identified, but it is not clear that they can be satisfactorily addressed unless 

effluents from full-scale raceways are used at the outset. 


Energy balances are needed for realistic conditions of operations. 


The project should address the potential hurdle of having an adequate supply of algae of various species, 

strains, and growth conditions. Capital and operating costs hurdles for commercial viability need to be 

specified.. 


Please refer the overall impressions section for the energy analysis and to the approach section for 

clarification on strain selection and relevance to populations harvested from outdoor ponds. 





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There is direct coordination if the operation fits into the overall process. Publications will result. 


The project began very recently. 


Early-stage relationship development with RAE (Jeff Kanel) for collaborative work on pre-concentration 

and filtration processes; Collaboration with SpinTek on embedded membranes for algal filtration. 

Collaboration with SpinTek and others on “Phantom” oil collection with nanofiltration; Development of 

additional partners (Graber, Koch, Pall, etc. as needed). 




Technology partners have been identified. 



Commercial interests of equipment manufacturers might drive tech transfer. The project can serve a 

watchdog function on claims. 


As pointed out, was have a number of partnerships and interactions with commercial entities that could 

help carry this forward. We have relationships with the major filtration manufacturers for food and 

beverage, mining, industrial fiber, and ink industries – each of which has a unique approach their specific 

industry’s needs. INL has relationships with companies that manufacture filtration systems such as 

Graber, Pall, Filtron, Parker, and SpinTek, and will utilize these resources as deemed appropriate. 






I believe this is a useful project. Confidence level would be higher if there was some support for this 

approach as a viable one in terms of the cost effectiveness. Something about the presentation left a strong 

impession of a machine looking for a problem, and here comes a problem with funding attached. 


The PIs may not be using realistic feedstock or suitable recovery targets in their harvesting/filtration 

research plans, but the project is in its early stages so these concerns should be correctable. 

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Is there a preliminary estimate of the cost per unit biomass undergoing this harvesting, dewatering, and 

drying process? The concern here is that the proposed process would prove to be utterly uneconomical. 



The researchers should begin by projecting the cost and energy consumption of these devices based on 

other applications. Then estimate the needed decrease in cost/energy to make algae biofuels feasible. How 

does that estimate compare with the best conceivable outcome from the research? This is an obvious 

starting point for any applied research. Presumably this exercise was done at the proposal stage, but it still 

needs to be shown to project reviewers. As it stands, these interesting technologies and the planned 

research could make progress for various industrial and water treatment applications. It is unlikely they 

will have a place in affordable algae biofuels production. Discuss this with one of the TEA teams and 

look in the TEA literature. 


My impression is that the presenter had a technology that "he can make work"for this particular 

application. Will this be an objective analysis? However, it appeared that some critical homework on 

energy estimates on spin-tech units had not been performed at the outset. Results will be of very limited 

applicability unless most of the tests use actual effluents from full-scale raceways. 


It appears to be a technology worth having a look at provided energy balances are executed. 


The project could benefit from definition of commercial performance criteria for various harvesting and 

dewatering scenarios. Process specifications should include feed stream characteristics, separation 

efficiency metrics, capacity requirements at scale, operating costs and capital cost. Characterization of 

algae slurries at various states of de-watering may be useful, especially rheological analysis. High internal 

moisture and high viscosities will be significant hurdles that can be partially defined by rheology. 


We appreciate the reviewer’s acknowledgement of the useful aspects of the project. We want to direct the 

reviewer’s attention to the statement early on in the presentation that this is an “analytic evaluation of 

cross flow filtration for harvesting/dewatering algae.” In that statement is the fact that there are a variety 

of cross flow filtration configurations each with its own inherent advantages, disadvantages, and 

limitations – thus the presentation of the levels of instantaneous face velocities in the presentation is an 

attempt to justify the fact that once we understand the mechanistic aspects of the membrane fouling 

caused by the presence of the algae, we will then be able to design an appropriate set of cross flow 

membrane unit operations to accomplish the task. Something that the literature has virtually no references 

that answer this basic question. At no time was it meant for the presentation to come across as a “machine 

looking for a job”. The spinning membrane unit is only a single component in the entire system that will 

be designed to dewater the algae to the economically viable level. Data for our preliminary cost estimates 

for processing 12 M gal water/algae were obtained through discussions with industry leaders (Millipore, 

Koch Membranes. Scenario 1) includes a floatation system and rotator system with 80% and 95% 

recovery rates, respectively and a conservative assumption that filtration membranes will be replaced 

every year. We anticipate an operational cost of $2.06 x 10E-5 per gallon for the filtration process or 

$246.90. Scenario 2) includes same assumptions as above but without the rotary membrane and assuming 

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conditions that prevent membrane fouling, the cost changes to $2.03 x 10E-5 per gallon for the filtration 

process or $243.80. No CapX were included in any of these estimates. We recognize that these 

preliminary calculations need significant refinement, but they provide an initial basis to warrant further 

investigation both from cost and experimental perspectives. Due to the combined complexity of the 

estimation and space limitations, we are open to discuss this further with reviewer. We do not fully agree 

with this comment that the presenter had a technology that “he can make work” for this particular 

application. Once again we apologize for any misperception that this is a technology in search of an 

application. We carefully evaluated costs for different filtration methods and came down with the two 

scenarios and further we provide the costs per gallon in the above examples which are surprisingly 

insensitive to method, but the level of solids (%solids) varies vastly ( Scenario 1 = 50 – 60%, Scenario 2 

= 15 – 20%) . We hope to consume much less than 15% of total energy, but need to optimize the 

debottlenecking separation of water and algae, thus the focused effort. Basically, it is not economic to 

filter all of the water in the raceway so a pre-concentration must be performed. There are three main 

approaches to pre-concentration: 1) full volume evaporation (used as a pre-concentration step much as 

does the salt industry, but the algae will probably eat up their stored oils to survive these types of 

scenarios thus rendering them nearly useless to us for oil production), 2) gravity assisted 

flocculation/settling, and 3) air (or other gas) assisted floatation. All of these methods are commonly 

utilized in industry (food processing, mining, etc.). We have selected gas assisted flotation due to its 

consistent results, speed, high concentration ratios, and energy efficiency. We are pursuing very smallscale 

studies with several varieties of algae to understand how the systems will perform before going to 

the expense of purchasing a full scale rotary membrane unit. We have read a number or TEA articles and 

will continue to consider the techno-economic feasibility of the cross-flow membrane technology. 

Project: 9.2.2.3 National Renewable Energy Laboratory 

Title: Efficient use of algal biomass residues for biopower production with nutrient recycle 

Presenter: Eric Jarvis 



Approach 5.43 1.18 7 

Progress 4.57 1.92 7 










The Problem is worthy of study. The approach is worth evaluating. Do not culture your own algal 

biomass! Partner with someone or get big quantities from actual industrial concerns. Improve the 

effectiveness of the approaches. Decide what the deliverables will be and design the program to deliver. 

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This project is considering the process of anaerobic digestion to change residual biomass to biogass 

(methane and carbon dioxide) and to recycle CO2 to cultures. Three species (Nannochloropsis, Chlorella, 

Phaeodactylum) are being used. This is good, because these algae are different biochemically while being 

good biofuels' targets. The PI promised to share data and the project's overall approach seems sound. 


Biogas production will be optimized for three algal species, best conditions will be scaled up, nutrient 

recycle will be explored, and results will inform TE and LCA models 




The goal is to characterize the digestion potential of extracted algal biomass and the nutrient value of the 

digested effluent. Unfortunately, the approach is includes the use of lab-grown algal cultures rather than 

biomass grown in pilot- or full scale systems. This seems both unnecessarily slow and potentially 

irrelevant (if the experiments need to be repeated to test biomass from full-scale systems). 


They are doing the "right" thing in terms of the subject area, and simply address each unit operation 

(growth, harvesting, dewatering, extraction....) but without new hypothesis to be tested, in the context of 

the long literature history out there. Also, AD of defatted microalgae biomass will not produce much 

methane (relative to CO2) unless one has added in carbon. 


It is highly desirable to define some specific quantitative goals based on economic and process 

perfromance criteria. Sourcing of biomass from a partner is a strong positive. 


We have chosen the three strains of microalgae for our studies (Nannochloropsis sp., Chlorella vulgaris 

UTEX395, and Phaeodactylum tricornutum) based on careful consideration. These strains sample the 

extremes of genetic diversity among eukaryotic microalgae being utilized for algal biofuels development. 

They are industrially relevant, but are also some of the best understood biochemically and 

physiologically. This provides several advantages; for example, our own work on the analysis of the cell 

wall composition of UTEX395 will be useful in designing enzymatic approaches to pretreatment. We 

have obtained Nannochloropsis from a commercial source (Seambiotic). Unfortunately, we are currently 

unaware of industrial sources for the other two strains. In-house cultivation allows us more control over 

growth conditions and will generate sufficient biomass for the studies. Of course, we would be very 

interested to hear of possible commercial sources for large quantities of biomass from these or related 

species. The criticism regarding a lack of hypothesis and context is noted, and perhaps this was a failure 

of the presentation. A number of technoeconomic (TE) modeling efforts, including NREL’s, have made 

assumptions about the efficiency of anaerobic digestion on oil-extracted algae. Essentially, the hypothesis 

that we are testing is whether those assumptions are reasonable. Andy Aden’s presentation, which 

preceded this one, covered analysis of the cost of algal lipids based on scenarios that include anaerobic 

digestion of algal residues after oil extraction. Inputs to the model (with current base case assumptions in 

parentheses) include efficiency of carbon conversion to biogas (65%), methane content of the biogas 

(40%), digester retention time (10 days), nitrogen recycle fraction (75%) and phosphorous recycle 

fraction (50%). Base case assumptions are based primarily on Weissman and Goebel (1996), and are just 

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that – assumptions. It is critical to establish whether these targets can be achieved, or possibly exceeded. 

Elimination of the entire anaerobic digestion component of NREL’s TE model (i.e., simply discarding the 

algal residue) results in a per gallon triglyceride cost increase of $2.22 for the base case pond conditions, 

or about a 25% increase in cost. Clearly, then, being able to meet these anaerobic digestion metrics is very 

important to achieving a cost-competitive process. The C/N ratio of the feed is one of the experimental 

variables we will be testing. As the reviewer points out, the methane content of biogas from de-fatted 

algae could be an issue, and one that we will be addressing. If carbon supplementation is required, it is 

critical that we establish that fact, as co-feed sourcing will have a significant impact on the economics, 

siting, etc. 







Not enough time under award has elapsed to gauge progress. 


The PI appears to be informed and reviewed appropriate background literature; this bodes well. 


The project is still in its infancy --preparation of materials for AD testing is just not nearing completion 




Progress has been limited by the difficulty in culturing sufficient quantities of algal biomass within their 

facility. 


Cannot judge - too little data. 


Project is in a very early stage. 


As the reviewers note, the project is still at an early stage. 




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This could be a relevant project. More progress and a sharpening of the project approaches would help 

this. 


The project is important to recycling and efficiency of energy extraction from algal biofuels. 


Commercial success of algal biofuels will require co-products of sufficient value and market size --AD 

for biopower has significant potential, but the details are poorly understood 




The project goals are certainly relevant to platform goals, but the approach unnecessarily limits the 

ultimate relevance of any results. 


The project proposed activities at each stage (such as enzymatic hydrolysis) without driving hypothesis 

anywhere or justification for a pathway that fits into the knowledge basis of the literature and reports by 

Benneman et al and Lundquist et al. 


The relevance is high because there is no scale limit on the concept. Detailed economic goals are needed. 


These comments are addressed by the discussion above under Project Approach. The “driving 

hypothesis” of the project is the validation of specific assumptions being made in the technoeconomic 

modeling, which are described above. The current knowledge base in the literature is a good start, but 

overall the amount of research reported on anaerobic digestion of algal residues is miniscule compared to 

the work done on other aspects of the algal biofuels process. 








assessments) 




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end-use) 




There is a reasonable initial description of these. 


Undefined animal waste is used as to innoculate the digester; will this pose replication problems? Will 

this process scale-up---and how will this be evaluated? It may be helpful to increase the AD efficiency 

with more initial proteases and cellulases. 


Need to demonstrate sufficient biogas yields from recalcitrant algal feedstocks, moderate capital costs, 

and efficient nutrient recycle 




Critical success factors were identified but no novel approaches were suggested that were likely to 

overcome these challenges. 


None stated to any real relevance. 


There appears to be very significant economic hurdles of capital requirements and of the low value of 

biogas and nutrients compared to the co-product value that will probably be needed to make algal fuels 

viable. 


The addition of proteases and cellulases to the digesters is an interesting concept, and we propose to 

address that through enzymatic pretreatment of the algal biomass before digestion. The longevity of 

enzymes added to the digester would likely be an issue, so we have opted for pretreatment in a more 

controlled environment. As stated in the presentation, our goal in this project is not the testing of novel, 

high-risk approaches, but rather the testing of assumptions for a supposedly “known” process. Having 

said that, we will of course be open to any novel concepts that might improve the yield of the anaerobic 

digestion process or the efficacy of nutrient recycle. It may be true that higher value co-products will be 

needed. This project and the associated technoeconomics will help establish whether or not that is the 

case. Note that some co-product scenarios (e.g., animal feed) might completely displace the anaerobic 

digestion component of the process, but others (e.g., specialty fatty acids, industrial proteins, etc.) will 

still leave a significant residue stream suitable for anaerobic digestion. 

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This cannot be assessed at this time. 


There is strong potential for good tech transfer from this project. 


Future work will generate vital data on AD of spent microalgae to gain value from both biopower 

production and recycling of nutrients. Data will be freely shared to benefit the entire algal biofuels 

community 




Technology transfer potential seems unlikely. 


Too early a project to comment. 


NREL work is widely shared. 


As noted, NREL work is widely shared and technology transfer will take place through publications and 

interactions with interested parties. 






Overall, I believe with some modifications, there could be useful outcomes from this project. 


Increasing the anaerobic digester's replicability with defined inputs and potentially adding additional 

sources of enzymes to break biomass should be considered. Recovery of energy from residual biomass 

after lipid extraction by anaerobic digestion is useful research. The diverse feedstock targets is a good 

aspect of the project. 

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Anaerobic Digestion : What are the anaerobic microbes involved? Please define them for the benefit of 

the review process. Is there progress so far in AD? The reviewer's recommendation for the PSI is to seek 

protein degradation (proteases) and cell wall degradation (cellulases) by which to pre-digest the biomass. 

Breaking up the biomass into the constituent components would substantially improve yields. 



The research goals are important and relevant, but the digestion results of future systems will depend on 

which algae are grown, the growth conditions, and the particular oil extraction method (current solvent 

extractions of all types have cost and other barriers). This project tries to cover the first two points by 

studying several species and growing them under conditions that might be used at full-scale. These are 

still somewhat speculative, but the oil recovery methods of the future are an even greater unknown. Thus, 

this project will be able to provide methane yield and production estimates for some speculative future 

conditions. However, even these preliminary results will be valuable. 

Presumably, the biochemical composition of the digester influent (and effluent) will be quantified and 

controlled, although this is not mentioned in the presentation. 

The emphasis on growing algae specifically for this project may consume most of the project effort. Only 

marginally sufficient biomass quantities might be produced using the small-scale culturing equipment 

implied by the presentation. Efforts should be made to identify another research group that is producing 

larger quantities of algae and extracting it and would be willing to share the residuals. The 2 kg from 

Seambiotic is a good start. 

Small, presumably batch, digestion experiments are planned for 12 months, prior to setup of 1-L, 

presumably semi-continuous-feed, digesters. Operation of only batch digesters for 12 months seems 

excessive because the more realistic data will come from the 1-L semi-continuous digesters, which appear 

to be scheduled to operate for only 4 months--a marginal amount of time to collect steady state data. The 

planned loading rate and C:N studies should not be the main emphasis because they are likely to give 

results similar to digestion of conventional wastes. Algae biochemical composition and oil extraction 

procedures are likely to be the dominant variables with respect to methane yield. The primary 

experiments should explore a range of these two variables. 

The algae regrowth work is valuable if light attenuation by the digester effluent is considered as one of 

the variables. 

What makes you think that the enzyme treatment will be affordable? 


The use of algal biomass from laboratory grown cultures drastically reduces the relevance of this project. 

Given the extensive knowledge base about anaerobic digestion, the funding level, and the two year 

project period, the goals appeared very modest. 


No real justification for any of the steps taken. 

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The project could benefit from specific quantitative goals based on commercial requirements for the 

process and the economics. The extent of the technology survey that has been completed by the team is 

not clear. 


Many of these comments are addressed above. Regarding the emphasis on batch digesters vs. larger scale 

semi-continuous digesters, that is a useful comment and we will do as much work at the larger scale as 

time and materials allow. However, our work at the smaller scale will provide much higher throughput 

and therefore the testing of many more variables. We believe that this approach will allow for better 

optimized digestions and maximize the output of useful data. The cost of enzymatic treatments, if they are 

shown to be effective, will be modeled. Recent advances in the production of low-cost enzymes for 

lignocellulosic ethanol processes suggest that such options may be affordable. We apologize if the 

presentation did not adequately convey the justification for this work. As noted above, when considering 

a process that currently is predicted to reduce algal lipid costs by $2.22 per gallon, it is important that the 

underlying assumptions be tested. Co-products are essential to an economically viable process, and any 

potential producer weighing the various options will want to make decisions based on actual data rather 

than assumptions. 

As discussed above, the possible need for co-feedstock will be an important outcome. We plan to use 

relatively defined co-feedstocks for initial studies (primarily cellulose), but may be able to test alternative 

low-cost materials. Regarding a question about recylcing of the AD sludge, we believe that sludge recycle 

may be problematic. Current scenarios focus on the liquid fraction of the effluent. Regarding a suggestion 

that algal residues simply be burned for power generation, anaerobic digestion for biogas/biopower has 

several advantages over a simple combustion scenario. In particular, the drying step required before 

combustion adds very significant costs and energy consumption to the process. Also, nutrient recycle 

would be more difficult; combustion of this high-protein feedstock would likely release much of the 

nitrogen as NOx, making recycle difficult and resulting in air pollution issues. Specific quantitative goals 

are discussed above under Project Approach. 

Project: 9.1.2.2 Sandia National Laboratories 

Title: Pond Crash Forensics 

Presenter: Todd Lane 



Approach 6.00 1.41 7 

Progress 5.86 1.36 7 








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The approach is reasonably good. How will multiple causes be addressed, or non-organic causes? What 

sort of modeling will be used. Many protocols need promulgation, i.e. sampling, storage, etc. 


This project began in October 2010 and is 25% complete. The project partners represent broad expertise. 

The goal of the project is "dipstick" 

development of rapid detection of common algal pond pathogens and detection of "novel" problems 

within several days by other molecular analysis (not detailed: Hi-Seq? Cloning?). This is an important 

problem and the overall approach is promising, including use of biotinylated DNA and subtraction 

technology with microfluidics to enrich for abnormal (pathogenic) sequences. 

The FY11 milestones are well defined. 


Develop methods to enrich for nucleic acids that are likely to derive from the etiological agent of the 

crash. Utilize ultra high throughput sequencing to identify agents. Create quantitative assays that facilitate 

the detection of agents at low concentration. Recreate the crash under laboratory conditions. Utilize 

advance spectroscopic methods to detect early hallmarks of algal stress. 


Promising concept, but it is not clear which types of organisms the process will be able to detect--viruses, 

predators, and pathogens are mentioned. Apparently bacteria, fungi, and zooplankton could be identified 

with this method in addition to viruses. It's odd that no particular known algae pathogens are mentioned. 

Non-crashing ponds similar to the subject crashing ponds should be tested as controls, to estimate the 

false positive rate. 


The objective is to identify agents responsible for pond crashes. The approach is to use high throughput 

sequencing to detect biological agents at low concentration. This approach seems technology driven 

rather than driven by the needs/experiences of managers of algal cultivation systems. 


Good work except that the justification was not made as to why etiological agents are the sources of 

crashes. What is the evidence justifying the funds to search here for the sources of crashes relative to the 

disapearance, for example, if micronutrients. 

In terms of looking at etiological agents, though, well thought out project. 


A good first step for this project would be to complete a failure mode and effects analysis (FMEA) on 

pond crashes to identify opportunities. If pond crashes have pathogenic causes with high impact, low 

detectability, and potential opportunities for mitigation, then the metagenomic analysis and development 

of advanced spectroscopy for pathogen detection would have clear value. leveraging previous work is 

highly desirable, but the opportunity needs to be better defined. 

It also appears that this project would be more efficient if it was initiated after the production system is 

better defined. 

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The proposed research has two goals. The first is the development of a system for the rapid identification 

of unknown agents by high throughput sequencing. The second is the development of a set of assays to 

detect the etiological agents, once identified, at low concentration (a "dipstick" assay for example). We do 

not propose to use high throughput sequencing as an everyday detection methodology as suggested a 

reviewer. 

There are several lines of evidence that implicate biological agents in pond crashes. The detrimental effect 

of protozoan grazers on mass algal culture has been well documented (Lincoln et al Aquaculture 1983, 

32:331-337 ). The role of other algal pathogens such as bacteria and viruses in production ponds has not 

be as well documented, likely because they are not readily detectable by microscopic analysis. However, 

in the study of natural blooms, there has been a number of recent reports of novel algal viruses and 

evidence of the important role that they play in the demise of a bloom (Lange et al FEMS Microbiol Rev, 

2009, 33: 295-323). In addition, bacterial (Mayali and Azam J. Eukaryot Microbiol, 2004, 51:139-144) 

and fungal pathogens Jia et al Ecological Engineer. 2010 36:1389-1391) have also been identified that 

affect algae viability and may limit or contribute to the demise of blooms . Given the fact that these 

agents impact algal biomass levels in natural systems it is not unreasonable to expect that they can have 

an affect in algal production ponds (Gachon et al Trends in Plant Science, 2010, 15: 633-640; Stephens et 

al Trends in Plant Science 2010, 15:554-564). The strength of the our approach is that it is not limited to 

detection of known agents. The methods for enrichment and characterization of nucleic acids from 

unknown etiological agents are applicable to the broad spectrum of potential agents (viral, bacterial, 

fungal, protozoan) and also to the detection and quantification of invasive or "weed" algal species. 

There are pond crashes that are induced strictly by the physiochemical parameters (pH, temperature, 

nutrient levels etc). It is also clear that there are pond crashes in which biological agents play a necessary 

(if not sufficient) role and that the effect of these biological agents can indeed be modulated by the 

physiochemical parameters. Fortunately many of the physiochemical parameters can be readily measured 

and it has always been the goal of this project to collect and include such data in our analyses. 

Unfortunately, the detection and quantification of biological agents which may play a role in pond 

instability is not as straightforward and this is the gap that this project intends to fill. It is our intent to 

study both crashed and un-crashed parallel ponds with the un-crashed ponds serving as negative controls. 

We will also utilize pre- and post-crash samples from the same pond as a source of controls. It is also a 

goal of the project to isolate, when present and possible, the etiological agent of the crash, recapitulate the 

crash, and to determine how physiochemical parameters modulate the ability of the agent to cause a crash. 

In terms of positive controls, there are several known algal pathogens (various phycoviruses, bacterial 

agents and predators (rotifers and other protozoa). Many of these agents are already in use in our 

laboratories for methods development and are intended for use as positive controls. 

We believe that there is insufficient data publicly available to carry out the failure mode and effects 

analysis, suggested by one reviewer. The current inability to detect and quantify biological agents (aside 

from grazers) precludes the determination of the fraction of pond crashes with pathogenic causes. 

Finalization of a number of protocols such as those for such as sampling and storage is an early research 

goal and we are converging on an appropriate solution. 

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Project is moving at an acceptable pace. 


The project has good partners and practical goals that are being approached well. 


• Developed and tested sample preparation and analysis methods • Developed host and background 

subtraction reagents • Developed agent isolation methods 


Some tasks have been completed. 


Progress is limited because the project just began (October 2010). 


New project so difficult to judge. Much more information could have been given (and defended) as to the 

development of the methods applied. 


It is early in project. 


The project team would like to have presented more technical detail but were limited by the required 

format and time allotted for the presentation. We have made significant progress in the development of 

positive controls and in house culture systems as a source of materials for analysis. We have obtained and 

are analyzing samples from an number of parallel pilot scale ponds. We are refining our methods to 

accommodate the use of frozen samples to ensure sample stability during shipment. 







The relevance is somewhat apparent if it is actually used to promote the scale-up of production. The 

investigators need to be sure they do not place hemselves in the position of promoting a methodology or 

technique and looking for a problem. Some partnering with pond operators is desirable. 

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This project has great relevance to algal biofuels' ponds at appropriate scale for commercial development. 

My only concern is what detection means for commercial facilities (e.g., would facilities choose to 

immediately harvest all algae and start over with new feedstock introduction after bleaching?) in practice, 

but it is unquestionably important to be able to assess pathogens quickly. This technique could also be 

applied to detecting other contaminants (i.e., possible pathogens or just competitive algae). 


Early detection will enable the “salvage” of biomass prior to crash Will facilitate pond remediation and 

return to production Will enable the testing of countermeasures against crash agents. Will increase 

annualized pond production 


Important topic 


This project seems to be driven by technology funded for bio-defense. The presenter failed to convince 

me how it is relevant for algal cultivation. 


Its fine to look at etiological sources but how is there affect going to be evaluated in terms of the other 

macro and micro environemental factors that could also be the cause? THis work should be put in the 

context of establishing controls (for one thing more information on the conditions of the ponds when the 

crash came....i.e. did it rain? Did the pH shoot up or down for some reason? Without this the wrong 

positive could be found. 


It's not clear how frequently RapTOR would be used in the management of a algae production system. 

The project did not present statistics on the occurenc of pond crashes from unknown pathogens. 


The team recognizes the importance of close interaction with pond operators. In the proposal phase we 

obtained letters of support from Sapphire Energy Inc and Dr. Ami Ben-Amotz (NBT, Seambiotic). We 

will redouble our efforts to form partnerships with pond operators early in the project. As part of this 

effort we have developed an aggressive communication plan to facilitate the development of such 

interactions. We agree that our methods under development can be applied to contaminants in general and 

we will work with algal producers to refine the concepts of operation for the technology. 

The PI acknowledges that there are pond crashes that are induced strictly by the physiochemical 

parameters (pH, temperature, nutrient levels etc). The evidence supporting the potential role of various 

types of biological agents in pond crashes has been detailed above. It is evident that there are pond 

crashes in which biological agents play a necessary role and that the effect of these biological agents can 

indeed be modulated by the physiochemical parameters. Fortunately many (but obviously, not all) of the 

physiochemical parameters can be readily measured and it has always been the goal of this project to 

collect and include such data in our analyses. Unfortunately, the detection and quantification of biological 

agents, which may play a role in pond instability, is not as straightforward and represents a blind spot in 

any cause analysis. 

We are concerned with sample transport. The choice of appropriate field preparation and transport is 

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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. 








assessments) 




end-use) 




These seem to be reasonably outlined. 


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. 


• Efficient and quantitative nucleic acid extraction and recovery. • Effective host and background 

subtraction 


Still needed: Determine false positive/negative rates for various pests. 


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. 


The project largely ignores other factors that can contribute to crashes that might have occurred. 

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Beyond the success factors related to identifying pathogens quickly, there is the factor of defining actions 

that could conceivably be taken on commerical scale. 


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. 







There should be a closer relationship with some pond operators. 


This project is early on but has high potential for excellent technical transfer. 


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. 


Collaborations planned are only related to sample provision. Can anything be gained from collaborating 

with others on the genetics or rapid detection methods? 


Technology transfer partners were not identified. 

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Too early to judge. 


Presentations, posters , and being tied into other algae efforts at Sandia show strong tech transfer. 


The project team is in the process of developing stronger ties with algal pond operators. During the 

proposal process we obtained letters of support from Sapphire Energy inc and from Dr Ami Ben-Amotz 

(NBT, Seambiotic). We are seeking to build upon these initial interactions and develop others. We are 

also in the process of developing collaborations in the area metagenomic analysis and have partnered with 

a team that has submitted a letter of intent to the JGI community sequencing program on this topic. 

Sandia has strong internal experience and capabilities in the area of rapid detection that the project will 

leverage. 






This is a potentially interesting project and approach. Successful implementation should be an aggressive 

goal for the investigators. 


This is an excellent project with important goals and well-defined milestones. Appropriate metrics (e.g., 

speed of analysis, sensitivity of detection, efficiency of analysis, common pathogens being analyzed etc.) 

are under consideration in the development of molecular diptick technology. 


Diagnosing the root causes of pond crashes at Sandia may provide some experimental results. The value 

of these results and conclusion would be limited by geographic and seasonal considerations. Meaning that 

microorganisms that feed on microalgae, the usual cause of pond crashes, would not be the same from 

location-to-location, season-to-season, or even year-to-year for the same location. Thus, the practical 

significance of the outcome from this effort would be limited. How could the PIs modify their effort to 

overcome this concern? 


Potentially valuable project using novel methods. The growth of in-house cultures will probably be very 

important to making progress. Samples coming from various field sites may be too variable and 

infrequent to determine crash patterns. 


This appeared to be a high-tech approach with limited applicability to real-world algal production. 

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I have conerns about the lack of positive controls in this project. Without those one can find exactly what 

he's looking for because of a lack of controls and a narrow focus on only one variable causing a crash. 


The potential value of the project is not clear due to lack of information on pond crash stastics and lack of 

definition of actions that can be taken on a commerical scale in response to pathogen identification. 


There are several published reports stating that predators can have severe deleterious effects on mass 

cultures. It has also been determined that viruses can lead to the demise of algal blooms in natural systems 

and that both bacterial and fungal pathogens can affect algal viability. It has also been acknowledged in 

the literature that, taken together, this this evidence strongly suggests that biological agents have the 

potential to severely impact algal production systems. It is also clear that the nature of these agents is 

poorly understood and there is a need for tools for the rapid identification and detection of these agents. 

This project will have two outcomes. The first is the development of a system for the rapid identification 

of unknown agents by high throughput sequencing. The second is the development of a set of assays to 

detect the etiological agents, once identified, at low concentration (a "dipstick" assay for example). The 

the proposed system for agent identification will not be affected by geography and seasonal variation. On 

the contrary, such an system would be universal and not dependent on any a priori knowledge of the 

etiological agents. Rapid agent identification will allow for the timely development of the simpler 

detection assays. An alternative strategy, that is under consideration, is the development of assays that 

detect general classes of threats. 

Whether an approach is high-tech or not is not relevant; a more salient question is whether the approach is 

cost effective. The same cost drivers that affect environmental and clinical sample analysis for biodefense 

are indeed relevant for algal production ponds. There is a need to minimize the cost of the initial analysis 

and the resultant assays. The high tech approach taken towards sample preparation and initial 

identification of pond crash agents will dramatically decrease analysis cost by minimizing reagent use and 

decreasing in the per sample costs for DNA sequence analysis. By increasing the signal content of the 

sample, we will decrease the amount of sequence data required to obtain a result, thus, increasing the 

number of samples that can be run in a lane of sequencing. 

It is clear to the research team that only a fraction of pond crashes will have a biological agent as a cause. 

It has always been the intent in the project to utilize un-crashed parallel ponds as a source of negative 

controls and known algal pathogens and predators as positive controls. Unfortunately the Principal 

Investigator did not communicate this effectively during the oral presentation. Once a putative etiological 

agent is identified, attempts will be made to isolate and propagate that agent and ultimately recapitulate 

the crash (fulfilling Koch's postulates). This process will have the effect of limiting false positives and 

identifying the abiotic parameters that contribute to the pond crashes. Metadata taken from the pond crash 

(temperature, pH, nutrient levels etc) will also be analyzed along with the biological signature to further 

understand the interplay between the agent and the environmental conditions. Identification and 

manipulation of the salient environmental parameters (with and without putative biological agents) with 

also facilitate the characterization of crashes that are due to solely to environmental stressors. However, it 

is clear that we can not, in the scope of this project, measure all environmental parameters, and the causes 

of some crashes will remain unknown. 

We agree that given the limitations in obtaining significant numbers of samples from crashed ponds, in 

house cultures will be important to making progress. We are modifying our approach to make more use of 

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this source of samples. We are also in the process of acquiring samples from closed photobioreactors and 

recycled ponds. We are in the process of developing collaborations with industrial partners that operate 

production systems. 

Project: 9.6.1.5 SRNL 

Title: Human Health Risk Assessment of Algal Production Systems: Toxins and Toxic Components, 

Harmful VOCs, Metal Speciation/Bioconcentration, and Pathogenic Microorganisms Associated with 

Large-Scale Algae Cultivation Systems 

Presenter: Chris Yeager 



Approach 3.14 0.64 7 

Progress 3.43 1.05 7 










The Approach lacks enough definition to support the importance of this program. If the issues are fully 

encompassing, how does the plan address them? What are the measures that should be employed, and 

how does the experimental plan address them? I see Phase 1 , but how do we decide Phase 2 and 

especially 3? The importance of the topic demands the best in terms of justification and explanation. 


This project includes collaboration between SRNL and TAMU-CC with the goal of examining toxins and 

toxic algae that develop in algal biofuels' raceways and ponds. The potential for algae causing these 

problems to be introduced regularly with commercial-scale culture of defined feedstocks appears 

unrealistic. Appropriate background experience in this area and strong knowledge of the rich literature on 

HABs was not evident from the presentation. 


Multi-dimensional; complementary aspects of lab and field work would combine geochemistry and 

biology. Overall objective is to provide experimental data for use in a comprehensive risk assessment of 

algae-based biofuel production systems on human health 


See other comments 


The goal of the research is to assess health risk associated with large scale algal cultivation and the focus 

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is on toxin production, VOC production, and human pathogens. The approach is sound but the toxin 

portion appears to have broad overlap with an existing knowledge base and research conducted at CDC 

and Woods Hole. 


One should start with a broad knowledge of pre-existing information. The CDC and the Centers for 

Oceans and Human Health have a lot of data already on this subject. This project should be linked with 

those groups. 


Because the geography, production system, strain, and mode of operation are not identified for 

commerical production, it appears that field measurements are pre-mature. 


While the structure of experiments can be outlined in advance (e.g. generalized cultivation processes, 

analytical screening methods) the full extent of experimentation will not be clear until Phase I, is at least, 

underway. It is our contention that these threats do exist – as per our preliminary findings. We believe 

that field sampling at active sites is the best way to perform the initial scoping studies required to evaluate 

if toxic metals, toxins, VOC’s and pathogens are of concern in these systems, and to best design the 

experiments to address the objectives of Phase II and Phase III. We will develop several risk scenarios 

that will outline how potential toxic exposures may occur to workers or consumers. A framework for a 

formal risk assessment will be developed and recommendations for further work will be made. The is a 

four-way collaboration: SRNL, PI Chris Yeager (project management, algae growth for Phase II, 

pathogen and toxin gene detection), NOAA, PI Peter Moeller (toxicity testing and toxin identification), 

Texas A&M, PI Paul Zimba (VOC analysis and outdoor growth experiments for Phase III), LANL, PI 

Jeri Sullivan (metal accumulation/cycling, toxicity) and Babetta Marrone (human cell toxicity). Dr. 

Yeager has 15+ years of experience using molecular techniques to detect microorganisms, including 

pathogens, in environmental samples. The number and type of pathogens that we will target obviously 

depends on the sample source (marine salt water, brackish inland water, wastewater, etc.) and location. 

We have spent considerable time determining which pathogens may be prevalent in different water 

sources and in different parts of the country. The list shown at the program review was simply a “working 

list”; different species/genes will be targeted depending on the sample source. The pairing of biology and 

water chemistry is one of the underlying strengths of this project. In terms of algae production systems for 

biofuels, very large scale, open air culturing will enable a number of environmental factors to act as 

modifiers (temperature flux, sunlight, airborne particulates, bacteria, fungal contamination, etc.) of water 

chemistry, and thus toxin production or the survivability of pathogens. Peter Moeller is the PI for the 

toxin work and is a PI at one of NOAA's Centers of Excellence in Oceans and Human Health 

(http://oceansandhumanhealth.noaa.gov/centers/). He was NOAA's scientist of the year in 2009 with a 

long history of work on algal toxins. Paul Zimba, Director of the Center for Coastal Studies at Texas 

A&M, is the lead for VOC characterization and has studied volatile metabolites produced by algae, fungi, 

and bacteria in aquaculture systems (http://ddr.nal.usda.gov/bitstream/10113/17447/1/IND44081503.pdf), 

has extensive experience with algal toxins. See comments above for Yeager’s credentials. This is a 

TEAM effort and the SRNL/NOAA/Texas A&M team formed (and was pursuing funding) before the call 

for this DOE program was released. Yeager is the lead-PI for this particular project because it was funded 

through a DOE National Lab call; however, all PIs are actively involved in the project. LANL was added 

to the mix because of their expertise with metals and human toxicity and their involvement with the 

National Alliance for Advanced Biofuels and Bioproducts (NAABB). Risk identification and avoidance is 

best and most economically performed at the beginning of a process, rather than after the fact. Analysis of 

field measurements is an essential element of our proposal. It is the variable geography, production 

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system designs and modes of operation that propagate the effective number of environmental cues, that 

are so often responsible for toxin production and pathogen survivability. We intend that experiments in 

Phase II and III help us better define culture and environmental variables that impact toxin and VOC 

production, the presence of pathogens, and heavy metal accumulation 







The project has been operational for limited time. 


The project started in September 2010 and is 10% complete. The lack of background knowledge of 

feedstock algae limits progress. 


PIs are actively conscripting sites for sampling and collecting data. Method development in the lab is 

progressing well, analytical data being analyzed 


New project; little basis for evaluation. Even weighting of scores in the average overall score require a 

low score here. 


Progress is limited because the project began in Sept 2010. 


But its a new project. 


Good progress in identifying potential risks. Getting meaningful information to quantify and mitigate 

these risk appears to be very problematic. 


We are fully aware of the common feedstock algae that are currently being investigated, but many 

companies are using variant strains, very novel organisms, or different phyla (cyanobacteria), and this 

information is not something that is intended for the public domain. Furthermore, it is not clear that the 

common feedstock strains are not capable of producing toxins. We have performed an exhaustive 

literature search and the team’s collective knowledge forms a very strong basis from which to proceed. 

It is beyond the scope of the project to completely quantify and mitigate the potential risks, but rather to 

provide the preliminary data required to enable assessment. This is the main thrust of our proposal. We 

have identified potential risks long before this project was funded, and there is a high likelihood that 

characterization of this risk will require novel approaches. Each problem (novel toxin isolation, 

purification and structural characterization; detection and quantification of pathogens in a novel 

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environment) has its own set of needs. We will develop analytical measuring tools such as MS, NMR, 

LC, DNA extraction techniques, Q-PCR, etc. to quantify these risks. Mitigation is very dependent on the 

threat. It is important to understand that in chemistry structure and function are intimately associated. One 

typically cannot mitigate toxicity until the compound responsible is characterized, detectible, and capable 

of being monitored. These are the products we are pursuing. 







The relevance is quite High. I believe it could be linked in somewhat better fashion to the experimental 

approaches and experimental outcomes. 


The potential for "rogue" facilities to develop the types of problems framed in the report is high, but this 

seems less so for the funded DoE projects. More importantly, other DoE projects presenting at this 

meeting had excellent command of pathogenic problems and were developing appropriate technology to 

detect them. It is important to be aware of the large literature, facilities, and resources available on 

harmful algae and the excellent and long-standing analytical techniques for their detection. 


Address questions of potential risk early rather than late; provide data for future risk assessment. This 

project supports the goals of the Algae Platform and OBP Program by: Addressing the social 

sustainability of algae biofuel production Understanding the potential negative human health impacts of 

algae biofuel production—from biomass production through harvesting 


It's important to look ahead to potential problems, but the state of algae biofuels development is so 

preliminary at this point, not much detail is warranted on human health effects. The inorganic water 

contaminants issues (eg, metals) would be good to assess in order to provide constraints to the current 

resource assessment projects. Potential use of pesticides/herbicides, as promoted by some groups, would 

be another topic to investigate to provide them with guidelines. 

The table listing pathogens and toxin producers is very excessive. Hopefully, this is showing a list of 

organisms standard to the the method (standard primers). Making special effort for most of the organisms 

listed (Slide 6) would be wasted effort because they would be no more likely to appear in algae ponds 

than other water bodies. 

This project should be a risk assessment desk study. Follow-up lab/field studies would expand knowledge 

in the areas with the most risk. 


Risk assessments are certainly relevant to overall program goals. However, it is unclear whether the 

results of this project will substantially contribute to the knowledge base. 

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It is relevent but does it duplicate ongoing work in other organizations and why is the DOE funding toxin 

work? 


A conceptual risk survey is important, but trying to measure and mitigate risks does not appear to be 

possible at the current state of technology development. 


The comments above do not apply to novel toxins or to “novel toxin generation’ by previously ‘benign’ 

species. I (Moeller) have worked in the HAB arena for over 20 years and I can assure you that there is 

little to no literature out there on toxins people simply do not know about. To apply current understanding 

of STX, PbTX, DA and/or microcystin to these systems is untenable. These projects require new 

approaches, must develop new detection and monitoring tools and just as importantly, but be reassessed 

routinely for even more “novel toxin generation”. 

We are attempting to obtain algae samples from a cross-cutting representation of current growers, not just 

DOE-funded projects. 

All other DOE projects investigating “pathogens” are concerned with algal pathogens (i.e. algal viruses, 

rotifers, etc.), not human pathogens. In light of the significant investments that DOE is making in algal 

biofuels R&D and commercialization, it is imperative for DOE to take a proactive approach to understand 

the risks. This includes the safety of products and co-products to consumers, as well as occupational 

hazards. If no early action is performed, responses after the fact are likely to be expensive and reflect 

negatively on the biofuel production community. We agree that risks associated with the application of 

pesticides/herbicides to algal growth systems should be assessed, but are beyond the scope of this project. 

We have spent considerable time determining which pathogens may be prevalent in different water 

sources and in different areas. The list shown at the program review was simply a “working list”; 

different species/genes will be targeted depending on the sample source. We are adapting pre-existing 

PCR methods into a PCR array to enumerate pathogens and toxin genes in algae samples. We plan to 

collaborate with Todd Lane, Sandia (also funded through this program) to perform deep sequence 

analysis on select samples. Between the team’s collective experience and an exhaustive literature search 

we have already conducted the “desk study”. The end result of that exercise convinced us that the 

potential for a human health risk is present in algae ponds. The initial scoping study (Phase I) is designed 

to provide the baseline data necessary to begin to carry out experiments (Phase II and Phase III) to 

characterize those risks. 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). Most of the toxin work will be performed at NOAA; however, this project does not an 

overlap with existing HAB research. An algal production pond and natural waters are VERY different 

systems. Nor does it overlap with work performed by the CDC. If we find strong evidence for the 

sustained presence of a human pathogen in any of the production system samples, we plan to contact the 

CDC and offer sample material for further analysis. One of the PIs (Zimba) has extensive experience 

working with the aquaculture industry and views this project as extremely relevant and unique. During 

the course of this project, we intend to collaborate closely with at least one algae production system using 

wastewater (we are currently talking with Patrick Hatcher, also funded through this program, of Old 

Dominion University). We can certainly learn much and apply existing data, techniques, etc. from the 

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wastewater. Yes, it is impossible to mitigate uncharacterized risks. This is the main thrust of the proposal. 

With new toxins/toxicity or the presence of human pathogens in novel environments, new approaches to 

identify the risk and detect the culprit must be carried out first. We will develop analytical measuring 

tools such as MS, NMR, LC, Q-PCR, etc. to achieve these goals. These tools, or adaptation of these tools, 

will represent a key asset to performing more formal risk assessments. 








assessments) 




end-use) 




The CSFs are the same as the other presentation on this Project. See the same comments. I do not think 

these adequately address the success of the project, which is of high significance overall, but requires 

some real assessment of what success means in the human health assessment. Input from EPA would be 

expected to be important. 


The project appears unaware of the NOAA Oceans and Human Health Centers, as well as the vast 

experience of the FDA and CDC in the stated area of research of the project. 


There is a need to establish the strains of algae to be investigated and justify the selection. Are the strains 

to be investigated the ones that would be used for biofuels production? If not, the work would find no 

application. 


See other comments. 


Critical success factors were identified but the plans to address a major obstacle (relevance of assessment 

results) seem inadequate. 


No real information was given as to the pre-existing knowledge of toxins to screen for, or as to the reason 

behind the screens chosen. This part of lacking. 

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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. 


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. 





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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. 


The potential for tech transfer from this project may be weak. 


Results will point to framework for formal risk assessment process, including recommendations for path 

forward 


Collaborators are involved which is good. 


Technology transfer efforts (outside of the risk assessment itself) were not adequately addressed. 


Too early to comment but the PI should partner with major efforts and groups. 


Unless or until the algae biofuel technology comes together, risk assesment will not be ready for external 

communication. 


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. 

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We will be laying the groundwork to enable a formal risk assessment, which will indeed require multiple 

agencies and much more work. 






This Project is important, and credit should be given for addressing it earlier than later. But the 

importance is not supported by the approaches and plans expressed. They need to be detailed and wellthought 

through to deliver impactful data. Input from other agencies is highly desirable. 


The expertise of the PIs may not fit the goals of the project. 


The entirety of this project appears to be the same as that by Enid (Jeri) Sullivan. Is the Algal Biomass 

Program funding the same project twice? Methods to be employed include DNA extraction and PCR 

array methodology for detection and quantification of pathogenic and toxin producing microorganisms. 

More information is needed to permit assessment of the applicability of this approach for the purposes of 

the program. 




The presentation failed to convince me of the value of this project. 


The project starts in the absence of proper incorporation of existing information. Without question the 

Centers of Oceans and Human Health have a plethora of information on this, as does the CDC. 


Maybe more could be learned from surveying risks in existing systems such as wastewater treatment or 

aquaculture, then trying to define and measure risks for an unknown system. 


This project is of poor quality due to the mismatched expertise of the PIs … See comments under 

approach and progress concerning the teams expertise. This project and Jeri’s are a single collaborative 

project. We each (SRNL and LANL) receive ~$400,000 per year for two years, but the project is 

coordinated by Sullivan and Yeager as a single project. Of the amount sent to SRNL, roughly 1/3 funds 

the NOAA effort and 1/3 funds the Texas A&M effort. The presentation failed to convince me of the 

value of this project. The PI was under the impression that the reviewers had access to the project 

proposals. Thus, most of the presentation was aimed at providing a VERY cursory review of the project 

and highlighting results to date and hurdles that we have encountered. This is a new research area that 

covers a lot of ground; we understand that it would be difficult to evaluate without access to the proposal. 

We have attached project overview/justification and Project Approach Summaries to assist further review 

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of the project. Without question the Centers of Oceans and Human Health have a plethora of 

information… No they don’t. Ironically, they come to us (NOAA) for what little there is on the new toxin 

producers and there is precious little out there. 

Must coordinate with the Centers of Oceans and Human Health. Coordination is already underway. 

However as pointed out earlier, if the DOE is to fund biofuel research, they are also responsible for the 

risk assessment. 

Why look at microalgae samples, as opposed to the water column. Very good question. We are looking at 

both. Cell mass and the culture medium (water) are matched during the sampling process. Both are 

extracted in identical fashion and worked up extensively. 

Very little known? Very little is known about the potential human health risks associated with algae 

production systems. Apart from Moeller’s lab there is no ongoing effort within NOAA to address novel 

toxins or toxicity. There is virtually no information on the topic that is germane to the biofuel issues. 

Not OK to just operate on the possibility that its there… We believe there is good reason to perform the 

stated research based on the existing body of knowledge and our collective experience. But the truth of 

the matter is – we don’t know what we will find. Isn’t that why we perform research? One might also 

argue that it is critical to operate on the possibility that it is there. For example, the literature that does 

exist provides many examples of toxicity being turned on/off for reasons few if any understand. The body 

of knowledge out there is incorporated in the PI’s experience in this realm. Novel toxins have no 

literature on them by default. Neither does their production, why they are produced, how they operate 

(mode of action), production cues, stability etc. – all unknown. Maybe more could be learned from 

surveying risks in existing systems such as wastewater treatment or aquaculture…. Good point. Algae 

ponds are distinctly different from wastewater systems and aquaculture. Paul Zimba has extensive 

experience working with the aquaculture industry and views this project as extremely relevant and 

unique. During the course of this project, we intend to collaborate closely with at least one algae 

production system using wastewater and will engage with wastewater professionals involved (we are 

currently talking with Patrick Hatcher, also funded through this program, of Old Dominion University 

concerning a collaboration in this regard with Hopewell's Regional Wastewater Treatment Plant). We are 

also planning to contact the City of Sunnyvale Wastewater Treatment Plant, which uses algae in its 

secondary treatment stage. We can certainly learn much and apply existing data, techniques, etc. from the 

wastewater field. 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). 

Project: 9.6.1.7 Los Alamos National Laboratory 

Title: Human Health Risk Assessment of Algal Production Systems: Toxins and Toxic Components, 

Harmful VOCs, Metal Speciation/Bioconcentration, and Pathogenic Microorganisms Associated with 

Large-Scale Algae Cultivation-LANL WBS#9.6.1.7 

Presenter: Enid (Jeri) Sullivan 



Approach 2.71 1.03 7 

Progress 3.43 0.73 7 


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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. 


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. 


Same as the project by Yeager. 


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. 


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. 


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)? 

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Because the geography, prodcution system, strain, and mode of operatin are not identified for commerical 

production, it appears that filed measurements are pre-mature. 


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). 

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The project has only been operational for a short time. 


This project began in September 2010, and the background knowledge of the PI should be 

strengthened(e.g., in literature review and consideration of appropriate and economical techniques for 

accomplishing goals). 




New project. No need to assess. 


Progress is very limited because the project began in September 2010. 


Results are very preliminary with only a few animal cell culture experiments over 48 hours or so. 


Good progress in identifying potential risks. Getting meaningful information to quantify and mitigate 

these risk appears very problematic. 



We have conducted an extensive literature review that is ongoing in the fields of biogeochemistry of 

algae, aqueous geochemistry in saline waters, algal metals uptake, and impacts of metals on algal toxin 

production. A review of the literature was not expected to be a part of this review process, however, we 

are happy to provide this information if requested. We also note the work of our co-PI Moeller (et al.) 

regarding toxicology and the interface of metals and toxin production in algae [7, 8]. We also note that 

the very deep cold snap in January, February, and March across all parts of the country caused many 

producers to temporarily suspend operations; those that continued were found to have cultures 

unacceptable for sampling at that time (dead algae or very low densities). We are working hard to catch 

up with our producers to obtain samples of algae and water now that the weather is more amenable. We 

appreciate the reviewer’s positive comment. We would like to note that this project is intended as an early 

screening of potential risks, not a full risk assessment. Please also view Yeager’s comment responses, as 

well. We believe that it is appropriate for DOE to initiate human health effects research aligned with its 

new focus on algal biofuels. This assessment is intended as an initial scoping study and, as such, would 

not be handled by either the CDC or Departments of Public Health at this level. If significant hazards 

(e.g., a significant pathogen) are found, the CDC will be notified. DOE has a long tradition of sponsoring 

human health effects research related to special DOE activities, such as nuclear weapons work. Babetta 

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Marrone (the co-investigator leading the human cell toxicity testing at LANL) conducted research on 

human health effects from beryllium exposure, as well as low dose radiation exposure. Both projects were 

funded by DOE. If our research indicates that there are health concerns, then we predict that NIH (most 

likely the National Institute for Environmental Health Sciences) would sponsor additional research, and 

that EPA would develop regulations governing worker exposure limits. The role of CDC would be to 

develop and disseminate health information and safety guidelines to workers in the algal industry, 

physicians, and the public. We believe that the DOE is performing a service to both state and federal 

health and regulatory agencies by being proactive in attempting to identify hazards that could be 

associated with their funded research projects, in collaboration with researchers who have considerable 

expertise in their fields. For example, Peter Moeller, our co-PI at NOAA, is an expert in algal toxicity, 

while many of the sites to be sampled are a part of the NAABB consortium. The deliverable of this 

project will be a report that describes our findings, and makes recommendations for next steps such as 

further research and cross-agency workshops to peer review our findings, and a possible framework for a 

risk assessment study of health impact from occupational exposure to algae in the context of the algal 

biofuels industry. Development of an actual risk assessment would most likely be a cross-agency activity, 

involving industrial hygienists, health professionals, industry, and scientists. (see section 6 below for 

references) 







The overall topic of Health Risk Assessment is highly relevant. The effects should be evaluated early on 

as well. The key is in selecting the right assessment techniques and criteria should be. The toxicolgy 

choices need to be explained as to the meanings of any outcomes. The selection of metals concentration is 

a good choice, but to re-study aqueous metal species chemistry is not worth doing. This information is 

well known already. To use XANES because there is an open available window on the beam line is 

unjustified. Give the slot to someone who really needs the time in the problem they are working on. 


Much is already known about the degree to which proposed feedstock algae accumulate 

materials/elements that are relevant to biofuels' facilities. New knowledge that will be helpful to biofuels' 

development requires close work with personnel on projects that are close to or at commercial scale. 




The project objectives are to identify potential human health risks (toxin and VOC production, 

human/mammalian cell toxicity, pathogens, metal accumulation/toxicity). What is the basis for thinking 

that any of these pose enough risk to warrant this sampling and algae production project? The study of 

culture media chemistry is worthwhile, but mostly from the standpoint of understanding trace nutrient 

availability and toxicity to algae. The level of risk to human health needs to be assessed in a desk study 

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(as mentioned in the companion SRNL review) before extensive sampling and culturing of algae is 

justified. 


Appropriate risk assessments will certainly contribute to the platform and program goals. However, the 

presentation failed to explain why a heavy metal risk assessment and mammalian cell toxicity assessment 

(and these particular methodologies) were relevant. 


I can see why one is worried about toxicity for operators, but why not just work through the CDC or any 

number of large Departments of Public Health? 


A conceptual risk survey is important, but trying to measure and mitigate risks does not appear to be 

possible at the current state of technology development. 


We appreciate the reviewer’s comment on the relevancy of the topic, and the need for early evaluation. 

We note that the beam time was awarded specifically to our student as a result of their successful 

proposal, and is an important part of their scientific educational experience. XANES Spectroscopy in this 

phase of the project was performed as a scoping exercise to determine its suitability in studying location, 

oxidation state, and presence of metal complexes in NS 1776 throughout a laboratory cultivation scenario 

using copper as a representative metal. XANES is used in toxicology as a method of risk assessment and 

forensic study of heavy metal toxicity and the presence of metal co-factors in a variety of systems 

(aqueous, soil, plant, and atmospheric particulates) [1-7]. It is commonly used as a complementary 

technique to methods like UV-vis, ICP-MS and Atomic Absorption Spectroscopy, all of which require 

destructive preparation. Most of these methods do not distinguish between oxidation state or different 

complexes of the same oxidation state, both of which are integral to toxicological studies [4]. Single cells 

are of interest because we wish to determine the interactions of metals with algae and not contaminating 

species. Cu was used as a test case specifically because it can speciate depending upon uptake and 

sorption conditions. Cu qualities in algal systems are well studied. The .1 to 10 mg/L Cu concentrations 

are representative of possible concentration ranges (including daily evaporation) in waters under 

consideration for algae cultivation. The study focused on the spatial distribution of copper, which can be 

absorbed internally by algae and/or adsorbed or precipitated on the cell wall. The location, oxidation state 

and speciation will affect bioavailability and toxicity effects depending on the mode of uptake 

(absorption, ingestion, and/or inhalation). Temporal measurements (1-4 days) were used to study the 

evolution of metal species throughout a growth cycle. For instance, in biological systems the location, 

oxidation state and speciation can be altered or transformed over time; a metal that is bioavailable or toxic 

initially may evolve into a harmless species or vice versa [5]. This is not a new field of toxicology; the 

application of known assessment and toxicological methods to these systems that have yet to be 

investigated in this manner is appropriate. Algae are known to strongly uptake and accumulate metals. 

When brackish and saline waters are used for cultivation, along with uncharacterized sources of fertilizer 

and mixed nutrient solutions, there is a possibility that higher than expected toxic constituents may build 

up in the system (water, algae, or sediments). One example is the elevated presence of arsenic in various 

forms in western ground waters [9]. Waters that are not used for human consumption may not be 

adequately characterized prior to use in algal cultivation. Understanding human cell responses by both 

chronic and acute exposure is of interest here; the possibility of critical concentrations of a metal or 

metalloid triggering a toxic reaction by the algae should be assessed. Other studies (including NAABB) 

have allocated extensive resources to nutrient availability and toxicity to algae, including work by 

Page 172 of 223




Sullivan and others at LANL now underway. A proactive sampling campaign is appropriate to allay 

concerns that have already been raised by both growers and the public, and to allow DOE to identifying 

potential risks and issues that may arise during funded research projects. (see section 6 below for 

references) 








assessments) 




end-use) 




I would like to see the CSFs include tthe critical planning of this key program, and the dissemination and 

application strategy. Where is the involvement of the EPA which will have the regulatory responsibility? 


A better understanding of biofuels' raceways /ponds would be helpful. Technology applied must be 

directed at useful application (e.g., most facilities would be close enough to a university to use X-ray 

microanalysis at a scanning electron microscope or just simple analytical chemistry (e.g., flame 

spectroscopy) for questions posed here; the investigators instead have used the SLACK facilities at 

Stanford. 






Critical success factors were identified. 


The criteria for toxicity and cell lines used were very poorly defined. 


It appears that bettter definiton of commericla algae biofuel produciton systems is a critical success factor 

that cannot be addressed in the timeframe of this project. 

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4. Critical Success Factors Results of the study will be made public through literature publication and 

reporting to the DOE. We expect that any indication of relevant system risks may then be taken up in 

depth by interested parties such as the EPA. Please see response to comments in sections above, 

particularly to comments in section 3: Project Relevance, and Section 1: Approach. We fully agree with 

the reviewers that growers should use nearby, readily available resources whenever possible. We do not 

recommend that the SLAC facilities be used for everyday analysis of water chemistry; the analyses that 

this facility performs will be used as appropriate in this study to answer topics relevant to speciation and 

subsequent toxicity. 







This program should have key interactions with others in the Biomass Program, as well as with EPA. This 

level of interaction is not developed in the review. 


This project would have more of a chance for useful tech transfer if the project were more sensitive to 

existing technologies and literature. 




Nice interface with other researchers. 


Technology transfer efforts were not adequately addressed. 


Not able to judge. 


Unless or until the algae biofuel technology comes together, risk assesment will not be ready for external 

communication. 


The project has extensive interactions with other institutions working under OBP, particularly NAABB 

consortium institutions. Because material transfer agreements are not in place for all participants, we have 

not disclosed all participants at this early stage of the project. In the future this information will be made 

available. Transfer of specific new technologies is not a primary focus of this project. Transfer of 

information related to risk is our focus here. However, LANL and SRNL, in particular have strong 

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technology transfer support in the event that transfer is appropriate. We agree that extensive concurrence 

between research and commercial entities is crucial for effective risk assessment. 






The Program itself is of high priority and relevance. It is a credit that it is being addressed earlier rather 

than later. Unfortunately the strategies and tactics are not well-developed in enough detail to allow 

appreciation of the importance. The gratuitous use of XANES is counter to the value of the project. 


This project has potentially important goals but needs to reconsider work in the context of potential 

problems mentioned above. 


The entirety of this project appears to be the same as that by Chris Yeager. Is the Algal Biomass Program 

funding the same project twice? 




The relevance of this project was completely unconvincing. My blunt impression was that this algae 

project was designed to support cell toxicity and heavy metal lab groups (rather than the other way 

around). 


This work is much better placed with the CDC or the Centers for Ocean and Human Health. 


Maybe more could be learned from surveying risks in existing systems such as wastewater treatment or 

aquaculture, thentyring to define and measure risks for an unknown system. 


Note this section includes references for the other sections. 

Please see responses to comments above regarding XANES. We regard the evaluation of this technology 

as neither gratuitous nor inappropriate. Please also see comments regarding Approach and Background 

from Yeager, as well as comment responses above and comment responses by Yeager et al. The project 

by Yeager and Sullivan is one single project. The separation of the project into two presentations appears 

to have been a confusing factor for the review panel. We would request that a single presentation be “tag 

teamed” in the future. Please also see comment responses by Yeager and Moeller, our Co-PI who is a 

member of the NOAA Center for Ocean and Human Health. 

Most interactions with growers are expected to be publicly available. However, specific agreements can 

be made (e.g. PII, NDAs) to allow appropriate review of future work. The data can be sent to reviewers 

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who are not made aware of the PI's and the site from which the samples are taken. We understand the 

concern of the reviewer. We hope that most, if not all, collaborators will work with us in an open process. 

However, various methods, including the use of PII, may be available to allow proper review of all the 

data by the reviewers. We agree with the reviewer and believe that both wastewater treatment and 

aquaculture systems are important and should be considered. More than one of our collaborators, and one 

of our PIs (Zimba), work in the aquaculture field, and we have already sampled at an aquaculture facility 

in Texas. References 1. Bresson, C., et al., A combined spectroscopic and theoretical approach to 

investigate structural properties of Co(ii)/Co(iii) tris-cysteinato complexes in aqueous medium. New 

Journal of Chemistry, 2007. 31(10): p. 1789-1797. 2. Isaure, M.-P., et al., Localization and chemical 

forms of cadmium in plant samples by combining analytical electron microscopy and X-ray 

spectromicroscopy. Spectrochimica Acta Part B: Atomic Spectroscopy, 2006. 61(12): p. 1242-1252. 3. 

Lin, H., et al., Speciation and biochemical transformations of sulfur and copper in rice rhizosphere and 

bulk soil—XANES evidence of sulfur and copper associations. Journal of Soils and Sediments. 10(5): p. 

907-914. 4. Werner, M.L., et al., Use of Micro-XANES to Speciate Chromium in Airborne Fine Particles 

in the Sacramento Valley. Environmental Science & Technology, 2007. 41(14): p. 4919-4924. 5. Gunter, 

K.K., et al., XANES Spectroscopy: A Promising Tool for Toxicology:: A Tutorial. NeuroToxicology, 

2002. 23(2): p. 127-146. 6. Wang, H.C., et al., X-Ray Absorption Spectroscopic Studies of As-Humic 

Substances in the Ground Water of the Taiwan Blackfoot Disease Area. Bulletin of Environmental 

Contamination and Toxicology, 2003. 71(4): p. 0798-0803. 7. Moeller, P.D.R., et al., Metal Complexes 

and Free Radical Toxins Produced by Pfiesteria piscicida. Environmental Science & Technology, 2007. 

41(4): p. 1166-1172. 8. Fisher, J.M., et al., Role of Fe(III), Phosphate, Dissolved Organic Matter, and 

Nitrate during the Photodegradation of Domoic Acid in the Marine Environment. Environmental Science 

& Technology, 2006. 40(7): p. 2200-2205. 9. Camacho, L.M., et al., Occurence and treatment of Arsenic 

in groundwater and soil in Northern Mexico and southwestern USA. Chemosphere, 2011. In Press. 

Project: 7.9.2.2 Desert Research Institute 

Title: Algal-Based Renewable Energy for Nevada 

Presenter: Chris Fritsen 



Approach 2.86 1.96 7 

Progress 3.71 2.19 7 










The approach is an unclear amalgam. Is the plan to use geothermal sources as farm ponds, or is the intent 

to look for algae that grows in various water sources and try and culture them for use in non-geothermal 

ponds? What is the intent of optimizing growth if there is no useful level of lipid content? How does one 

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start from zero knowledge base in terms of this problem, and the potential bioproduct spectrum and make 

progress worthy of the expenditure? A very poor discussion of the approaches. 


This project's goals are to develop algal biofuel feedstocks that are appropriate for growout in geothermal 

waters in Nevada. DRI, AG-Energy Corp and Univ. Nevada-Reno are involved. A GIS database has been 

assembled and T, pH, and conductivity of geothermal ponds are being assessed while Nile Red staining of 

algae from geothermal ponds is beginning. The project plans to develop a raceway and look at C 

utilization of selected algae. It is not entirely clear why the scale of geothermal waters' characterization 

being undertaken is needed and how the algal strains collected from these ponds will be compared, 

including to non-geothermal feedstock candidates. 


The project will bolster and initiate basic research regarding the development of algal growth and 

production systems in conjunction with geothermal resources throughout Nevada. 


Project is an exploration of algae cultivation potential in Nevada, in particular using geothermal waters as 

media. The alternative, use of geothermal waters to warm media (as done in previous Nevada-based 

research), does not seem to be considered. 

The project lacks a vision of the design and requirements of future algae production facilities in Nevada. 

One of the goals is to assess the scalability. The researchers need to define an envisioned scale-up facility 

design in order to complete this goal. What is the fate of the geothermal water after algae has been 

grown? What pond temperatures will be achieved? What nutrients will have to be imported? How will 

these nutrients be recycled? etc. Focusing only on the biological performance of strains will not allow this 

task to be competed. 

Most algae collected from the environment will not be suitable for a cultivation environment. The 

planned use of outdoor ponds will be a benefit. Targeting of only a few sites may allow this project to 

develop methods to cultivate a few strains. This would be a good accomplishment. Any on-going search 

for ideal strains should be minimized and instead focus on a few strains and waters. 

Carbon is a major limiting factor in commercial algae production. Dissolved inorganic carbon and free 

CO2 availability at the sites should be evaluated along with the other chemical characteristics. 


The project approach included traditional methods: screen strains, examine beneficial uses, and then 

outdoor scale up. There was no justification for why this approach would succeed in light of past failures 

of similar approaches to yield strains that thrive in outdoor production systems. 


If the project is about the use of geothermal waters in Nevada, than the entire project should focus on the 

devleopment of that research interms of ability to create ponds and such that can cultivate wild consortia. 


A viable process concept for using geothermal water to control the temperature of algae production 

sytems at meaningful scale needs to be developed. 

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We recognize that work on pond scaling and production systems has been somewhat extensive. However, 

attempts to achieve consistently high productivities were often hampered by low temperature conditions 

encountered (Roswell, NM). Thus, it was concluded that some form of temperature control may well be 

required (page ii, Technical Review; Sheehan et al. 1998). Prior works and analysis has shown that the 

Great Basin (Nevada being the major area within the greater Great Basin) could be a favorable location 

for systems that rely on solar radiation (http://www.nrel.gov/gis/solar.html). Therefore research on 

developing pond systems (with local algae that can actually grow in the water) in the region makes just as 

much sense as other locations that have been proposed. However, in addition, gaining temperature control 

of production systems in these locations and extending the growth seasons also has been recognized as a 

key component to allowing the viability of the pond-based production facilities (Sheehan et al. 1998 and 

references therein). The Great Basin has a multitude of geothermal features and resources with potential 

to be used to provide energy in the form of geothermal power generation. At the back end of the plants 

used to generate electricity, geothermal water is “wasted” at temperatures that are not conducive for 

electricity generation but still are well above 70 deg C (with the volume of flow and rates depending on 

the generation plants). However, even small-scale turbines can produce 500-900 gallons of water per 

minute at temperatures exceeding 70 deg C (some lines produce at temperatures at or near 100 deg C). 

Even at these low rates of flow- temperature control of pond systems at 30-50 deg C can be achieved 

either by direct addition or by a combination of direct addition or thermal insulation systems (the "waste" 

energy nominally represents over tens of megawatts of heating potential). In addition, there is even more 

untapped dry-based geothermal reserves that may be watered for energy production in the future. Most all 

geothermal systems have the water being re-injected after use and all notional designs for these systems 

would have the waters being returned. The reviews were critical of the fact that this program was not 

directing the resources for the design specifications of plants for the geothermal control. The private 

sector and other design engineers are doing some of this work and will undoubtedly do this work in 

earnest once it is shown that the hard work of achieving reliable biomass production at higher 

temperatures is completed and the other critical issues are dealt with (such as stability and nutrient supply 

etc...). It seems the scale of energy supply from heating should be better communicated in the future and 

we regret that some on the review team had not the grasp of the amount of potential energy that exists in 

these systems and this was not articulated well at the meeting. 

Efforts will take into account the well founded comments regarding C02 assessments and efforts to focus 

on only a few of promising strains and consortia rather than expending all the resources searching for the 

"ideal" strains. 







Activities are underway, but they seem disjointed and not well-rooted in an end objective. 


This project is about 15% complete and more focus on practical targets might be helpful. The work seems 

too much like an ecological characterization of geothermal waters vs a focussed study of whether 

feedstock algae are to be found there. 

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The project is progressing as indicated in the project management plan. Field sampling and laboratory 

experimentation will continue. 


Water chemistry at some sites has been characterized. Some algae samples, presumably from geothermal 

sites, have been characterized in lab for growth and lipid content. Amendments to geothermal waters have 

been developed for algae media. A needed safety plan is in place --good. 


Progress is limited because the project started in September 2010. 


Only a few microalgae reactor cultures and developed the Nile Red assay. 


Technical progress is hindered by lack of conceptual temperature control ssytems supported by 

engineeering calculations on heat and water balance in potential algae production systems. 


We agree that temperature control can be developed more as the program progresses and the scale of 

water production and processing becomes known at sites. However, the general approach is such that the 

private sector or other RnD activities will take this and other aspects on once it can be shown that biomass 

can be reliably produced at desirable rates. It should also be noted that some or much of the nutrients can 

be gained from the geothermal waters themselves (several sources have phosphate concentrations in 

excess of 10 uM and nitrate and ammonium concentrations on the order of 500 to 3,000 uM) and this key 

aspect of development relies on gaining better and up to date water chemistry information. Moreover, 

much of the prior works in the field targeted developing strains in only a few types of reproducible 

defined media. The RnD undertaken here is site-directed work and aims to work on only a few of the 

waters and make growth media specific for the sites. The geochemical interactions of the water with 

nutrient additions is showing that seemingly simple nutrient additions or water modifications are not 

without issues that should be addressed. 

Work on bioreactors or open cultures in the lab and in greenhouses provide a means to gain results 

without having to expend large amount of the resources in testing strains and consortia in the field (which 

is much much more expensive). The scaling approach is one that has its challenges. Yet, the approach of 

developing consortia and or strains in the site-specific water is more likely to succeed when compared to 

prior works that tried to gain isolates on defined media not tailored to sites or the approach that was 

suggested of just building a pond and seeing what grows. 





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There is little development of relevance of this project. Assuming some aspect of geothermal energy is 

the basis, what is the relevance? Is it the creation of new organisms, is it the use of geothermal waters, is 

it the application of extremophiles under average conditions? Other work has clearly been done on 

extremophiles, but what lessons were learned that drive a new project here? 


This project would be most relevant to use of raceways associated with an industrial process that 

produces warm waters with similar physical characteristics to geothermal waters in Nevada. Whether any 

strains isolated there are novel relative to lipid concentration etc. of existing feedstock targets is unclear at 

this point. 


The project contributes to meeting the platform goals and objectives through the potential identification 

of a sustainable, high quality feedstock supply from a geothermal source, plans for development of 

logistical systems to meet physical and quality specifications through analysis of lipids produced and 

investigates the development of a commercial-scale supply system with planned optimization of a pilotscale 

facility. 


The project needs to use existing information to establish a preliminary resource potential for algae 

products in Nevada. Without an idea of how much algae could be grown in theory, the relevance is 

unknown. 


The relevance of the project to program objectives is in its stated goal of utilizing local resources for 

bioenergy production. However, the approach and focus of the project are likely to limit its contribution 

to program goals. 


The majority of work presented is a near duplicate of work in the other consortia and projects. 


Screening and testing thermophilic algae holds little relevance if a potentially viable high temperature 

production system in not conceived. 


Based on the general program’s targets and a number of key recommendations from the prior works of 

several ASP researchers (e.g. Bennemann, Cooksey, Sheehan, Pienkos etc…) and others regarding the 

barriers and approaches that should be addressed to get to the program’s targets- the overall approach was 

developed. The approach is indeed often similar to that of the much larger consortia that are funded 

within the DOE program (e.g. the NAABB) and includes developing research that is called for within the 

DOE program. Such research includes the following: providing site-specific information regarding water 

suitability for culturing, gaining isolates and/or consortia that can grow in site-specific waters, enhancing 

the quantum yields of photosynthesis and production systems (e.g. Weyer et al. 2010), gaining 

temperature control to sustain maximum production through cool periods (Sheehan et al. 1998), and 

increasing temperature tolerance (e. Sheehan et al. 1998, Weyer et al. 2010) such that algae ponds may be 

more cost effective. Doing such work should yield the information desired to gain production rates on the 

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order of 10-30 g/m^2/day throughout the year (which from a purely quantum yield theoretical calculation 

can be achieved in approximately 4-6 hours of daylight in the Nevada irradiance regimes given 

temperature is not limiting). Despite some reviewer’s comments suggesting that this work is repetitive 

and all of this work “has been done”, it remains that site-specific work and work relevant to production 

systems at enhanced temperatures has not actually been completed. There is presently little to no attention 

being directed at thermal systems and there remains good potential for key developments in this area. The 

reviewers are correct in that lipid research in hot springs has been “done” but this work has mostly 

targeted the cyanobacterial mats and prokaryotic components therein that do not necessarily produce 

energy storage compounds or lipids in amounts that are deemed desirable targets for the DOE biomass 

fuels program. Also- much of the work on lipids in hot springs has targeted the very warm and 

thermophilic cyanobacteria that grow at temperatures approaching ca. 70 deg C. Since photosynthesis is 

hampered at these temperatures, research and opportunities for key discoveries remain within the realm of 

30-70 deg C (e.g. in the area of achieving desirable quantum yields of photosynthesis for rapid biomass 

production at lower standing stocks). We recognize that it is surprising that there remains much work to 

be completed on microalgal production and bioengineering of systems in the range of 30-70 deg C. It is 

intriguing that several programs did pursue this research early in the ASP days. However, it appears that 

thermal control was not achievable at some of the test sites (freezing temperatures at night) and attention 

was direct to seeking cold-tolerant strains and systems (page 20, Part II: ASP Technical Review; Sheehan 

et al. 1998). The challenges in cultivation at higher temperatures are not trivial and it may have been that 

culturing at low temperatures was completed in the early phases of this program, yet we too feel this work 

should be progressed. 








assessments) 




end-use) 




The listed CSFs are not really success factors for the delivery of a valuable contribution. When asked how 

the presenter defines successful conclusion to the project, there was no cogent answer. Since the basis for 

this work was not well expounded, it is not surprising. 


Milestones were not entirely clear; focus on rapid screening of new strains that may be novel relative to 

lipid production is important. More consideration of background literature on geothermal source algae 

(e.g., Italy, California, Montana....etc.) might improve this project. 

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Critical success factors which will define technical and commercial viability are the identification of 

several candidate algae (strains and/or consortia) capable of rapid growth/lipid production in conjunction 

with Nevada’s geothermal resources 


Inadequate as described in the presentation. See other comments on need for an initial scale-up facility 

design. 


Critical success factors were identified. However, plans to address these obstacles were inadequate. 


None were present. 


A critical success factor not identifed is definition of a process concept for keeping the water hot. 


The critical success factors outlined in the presentation which will define technical and commercial 

viability were presented as the identification of several candidate algae (strains and/or consortia) capable 

of rapid growth/lipid production in conjunction with Nevada’s geothermal resources. Though the 

presented success factors lack specificity and do not directly reference the productivity improvements that 

are anticipated to be attained through the project, success in the stated factors will provide a valuable 

contribution to the program. Among the R&D challenges outlined in the Technology Goals of the 

National Algal Biofuels Technology Roadmap (page 6) are obtaining sample strains from a wide variety 

of environments for maximum diversity, investigating multiple approaches to algal cultivation, and 

optimization of systems for algal productivity of fuel precursors (e.g. lipids). A successful project will 

contribute directly to these stated challenges/goals. In hindsight, we would amend the stated critical 

success factors to include such specific gains that we expect to achieve from the project. Some of these 

critical success factors for the project include: The identification of strains and/or consortia associated 

with geothermal waters exhibiting capacity for annual average biomass production values > 30 g/m^2/day 

with >30% lipid content on a dry weight basis as outlined as a target in the Roadmap (page 81) at 

elevated/controlled temperatures; The demonstration of the potential for scalability of at least one of these 

strains or consortia through production at the stated rates in an outdoor pilot scale test facility for greater 

than 1 month (thus addressing both the rate and stability issues); To assess the utility of and provide the 

opportunity for technology transfer of algal production in association with geothermal waters through the 

development of pilot scale test facility using geothermal waters for heating and/or growth media. 

The success factor, of providing the design specifications for temperature controls of these systems, was 

not built into the program as a desired target- mainly because there are several private-sector firms that 

are doing this type of work. Simple calculations outlined previously show the heating potential is on the 

orders of tens of megawatts even using the waste heat from the small electrical plants and demonstrates 

the potential viability of the resource. However, the overall viability of these notional facilities will rely 

more on the sustainability of biomass generation and water use and not likely temperature control given 

the extremely large heating capacities of these resources. 

Page 182 of 223










There is little acknowledgment that the rest of the world has made much progress in these areas, and that 

the basis for judgments need reference to this other body of work. This seems a project in a vacuum, with 

undefined objectives. To expect to start a program which will be optimizing a pilot operation to deliver 

valuable (undefined) products in a few short years is not believable. 


The potential for tech transfer is unclear at this point. There is also some concern about the 

characterization of strains (molecular ID needed). 


Unclear at this point. Needs to be clarified. 


The research team is all at UNR/DRI, plus a company. The skills of the team are not described. Seems 

insular. 


Technology transfer efforts were not addressed. This appears to be an effort entirely focused within 

Nevada. 


Work is being done in an absencea of the large microalgae community. 


Colaberation with engineering expertise in algae growth system design, including heat and mass balance 

modeling is needed. 


The work is not being done in a vacuum. Rather the work is being conducted as the scientific community 

has called for in several recommendations by national teams (e.g. see the National Algal Biofuels 

Technology Roadmap) and much of the work is synergistic with the larger consortia's aims. The work that 

targets technology development downstream of the production systems is being covered in other works 

within the DOE program (e.g. the NAABB, etc..) and throughout the nation. Thus, this particular 

grant/program has not targeted many of these items. Rather, the work does focus on RnD to gain 

production systems at relevant temperatures (ca. 30-60 deg C) that are not necessarily just relevant to 

Nevada- but are relevant to the Great Basin and any other area where the nexus of water, solar radiation 

and temperature control can be gained and the algae can be developed to grow within the site-specific 

waters. 

A small component of the program has been set aside to look at alternative uses of byproducts. The team 

recognizes that this work does not necessarily address one of the program's highest priority a critical 

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success factors and we may revisit directing resources in this direction. 

For information on the labs research programs, see http://www.dri.edu/chris-fritsen. In addition, Dr. 

Fritsen has worked in the ASP years ago in Dr. Cooksey's laboratory, has extensive culturing experience 

and knowledge of nutrient and photophysiological attributes of algae. He has experience in systems 

designs and program management through work on NASA programs as well as the polar research vessel 

committee service. Additionally, co-investigator's have requisite experience in analytical capacities to 

support the efforts (e.g. lipid analysis, water chemistry, microscopy, etc...). Additional team members 

from UNR and AGEnergy have expertise in private-sector partnerships and algal culturing as well. 






The weaknesses have been enumerated already. The presentation did not show thoughtful objectives and 

scope. Nor did it show a relationship with all the other work in the algae bioproducts and even 

extremophile areas. Why would one consider scale up without any technoeconomic reason to do so? The 

questions about this project are multitudinous. The DOE officer should be demanding a well-reasoned 

scientific program. 


This project appears to be a basic study of geothermal algae in Nevada; it may reveal something 

interesting about these algae, but it is important for the PI to focus on development and application of 

information to commercial biofuels' production needs refocus. 


This is a somewhat unique and worthwile effort seeking to apply basic research regarding the 

development of algal growth and production systems in conjunction with geothermal resources 

throughout Nevada. 




This project appears to be more focused on capacity building within Nevada than on addressing Biomass 

Program goals. 


Much of this work is being done by the consortiums and the focus should be on geothermal issues of 

production. 


Additional concept development for high temperaure production systems is needed. With several day 

growth cycles and thousands of acres of growth system area, temperature control appears to be 

problematic. 

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In brief, we fully intend to continue to work within the national program’s framework as outlined by the 

2010 report, in collaboration with colleagues in large consortia and to remain synergistic with the large 

consortia’s and the private sector’s efforts. That said, however, we also recognize the value of the model 

that DOE has worked under in the past whereby independent research approaches and lines with similar 

desirable outcomes has the potential for independent discovery and technological breakthroughs. The 

work is indeed unique and being conducted in a lab and area where the resources to do so are available. 

The other misplaced reviewer's comments are indeed misplaced. The work is not just relevant to Nevada 

(see section on relevance) and the scaling for technoeconomoic analysis can and should be done. 

However, if it cannot be demonstrated that biomass production can nominally occur in these waters then 

such technoeconomic analysis will simply lack a basis for speculation or analysis. The scope of the 

research is seemingly wide and ALL of the potential factors that may inhibit the development of algae in 

these areas is understandably not addressed (e.g. harvesting, drying, transport etc...). Yet, the RnD does 

only target work in regards to geothermal water, consortia and strains for development. The work does 

not encompass the scope the NAABB encompasses. Yet understandably, the field does have standard 

procedures and approaches that are utilized throughout the discipline field. It should not be surprising that 

there are similar activities and approaches among and between labs. 

We will work to better provide a clearer assessment of the the overall scale for thermal-algae systems in 

the Great Basin. However, with over 21 operational power production facilities in Nevada alone and more 

in development (e.g. http://www.nvenergy.com/renewablesenvironment/renewables/geothermal.cfm), the 

potential marriage of the uses of the waste heat and growing algae therein is real. Additionally, waste heat 

from other power plants and the specific waters that accompany these plants likely should be investigated, 

yet lie beyond the scope of the present study. 

The peer review does appear to have some issues, given that during the preparation of the presentation 

materials we were given instructions to provide materials for "high-level" program synthesis and reduce 

the amount of technical details-- Yet, some/many of the reviewers comments seemingly were directed at a 

much more technical level. 

--------------- 

Project: 7.7.2.18 Brooklyn College 

Title: Development of Pollution Prevention Technologies 

Presenter: Juergen Polle 



Approach 4.33 0.47 6 

Progress 4.67 1.49 6 






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The Approach has several aspects. Basically the investigators are intent on establishing reliable, analytical 

methodology to determine full lipid composition profiles in timely fashion (one hopes) to replace the 

qualitative nile red stain test. Now if that could be done it would be a useful contribution, especially if it 

were a short, inexpensive, low manpower analytical method. There probably needs to be a diversity of 

samples run through to determine the variations. Good enough. It is when the project direction indicates a 

prediction is to be made on the best choices of algae that I believe the approach goes astray. How do we 

decide good, better, best? There is a claim to evaluate 2000 strains of algae. Is this logical or desirable? 

First, some of this must already be done. Second, is a scientific selection a better approach? 


The approach is to isolate new strains of algae that might be useful as feedstock; this goal is being met by 

isolation of 3000 "strains" from extensive collecting, including in Utah, of freshwater, brackish and 

marine strains. Analytical methods such as MS, LC are being used to define lipid concentration, which 

the PI considers important compared to Nile red staining. It was unclear that the collecting effort was 

focussed on scientific questions related to habitat that might result in the highest possibility of useful 

novel strains with high lipid accumulation. I do think some collecting of new strains from some habitats is 

likely to be fruitful to commercialization, because this may eliminate the need to pursue GMO strategies 

for high lipid production. However, Martek etc. have many good lipid producers; will there be any 

comparisons with the "best of show" to date? 


For strains sampled from the general environment such as these, lipid content and lipid characteristics are 

likely a weak indicator that the strains are suitable for mass cultivation. 200 strains have been 

characterized; now focus on determining whether any of the leading candidates can be grown outdoors. 

That experience will help in the planning of any further screening. The delivery of the strains to NAABB 

or others with outdoor facilities should be done quickly with the outdoor results feeding back to the 

Brooklyn College strain work with as quick a turn around as possible. 

The catalyst research cannot be evaluated with the small amount of information provided. 


Since the presentation only focused on the strain screening project, this review only covers that section of 

the project. The stated objective is high throughput screening for lipid producing strains and 

characterization of lipids. However, in light of the failure of previous algal screening efforts in yielding 

strains that successfully compete in full-scale outdoor raceways, there was no justification regarding why 

this particular effort would be more successful. The main purpose of the project was for capacity building 

for the college (purchase of equipment, hiring of personnel, training of students). 


What is the relevance to just screening strains? There are several sites that do this, for example UTEX. 

Why is this needed over existing collections? 


Validation of the screening system is needed. 

Page 186 of 223




An experimental design to determine interactions between the parameters used in the growth system and 

strain responses may have value. 


For clarification for the reviewers, the PI would like to mention that the strains that are being screened in 

this project were isolated in a previous isolation effort. No new isolation will be done for this project. The 

screening effort ongoing in this laboratory is not simply an isolated project. Some strains that were 

isolated and passed the first Nile Red screen in this laboratory in the past were already passed on to 

collaborating laboratories for testing in larger systems. The ongoing project is meant to replace the Nile 

Red screen with a more effective screen that will be faster and more accurate. Regarding comparisons to 

'best of show' to date, the production strains Nannochloropsis salina and Nannochloropsis OZ1 are 

currently being used in this laboratory as 'bench mark' strains for lipid productivity in side-by-side 

experiments to the novel strains identified through our screening. 

This PI begs to differ in opinion from the reviewer in that strains from culture collections have been 

successfully tested in outdoor test beds. Therefore, the statement of the reviewer is not exactly accurate. 

However, due to confidentiality agreements this grantee cannot disclose the nature of those tests. As an 

answer to the comment that validation of the screening system is needed, it can be disclosed that two 

candidate strains identified in the previous sampling/isolation/screening effort of this laboratory were 

already grown repeatedly by a collaborating laboratory in the southern United States over several months 

in a greenhouse at a scale of about 30 Gal. Both strain have performed well and further tests are currently 

ongoing. Moreover, more strains will be cultivated in testbed facilities of the NAABB very soon. In lieu 

of the absence of any truly large-scale raceway system within the USA, it is this PI’s opinion that at this 

time the results of such experiments, which are not part of this research project, may serve as ‘proof-ofconcept’ 

for the potential of scale up for strains identified in the PI’s activities regarding strain 

isolation/screening in this laboratory. 







Progress is apparently adequate to date, given the project start date. 


Most of the work to date of this project that started in September 2010 is collecting effort. 


200 strains evaluated. Enough for now. Learn more about the best strains identified so far. This culture 

work can be done at Brooklyn College as well as the NAABB partners. 


The project has made progress toward its stated objectives (purchase of equipment, hiring/training of 

technician, initial strain screening). 

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It is a new project. 


Earliest possible test of the screening procedure against longer term outdoor cultivation resutls will help 

define progress. 


Again for clarification, this project does not include any strain collection and/or isolation efforts. Strains 

to be screened already exist in the laboratory. At this time, the PI is ‘learning more about the best strains 

identified so far’. In a related project that is not part of this screening project, candidate strains are being 

tested in the laboratory all candidate strains that pass the screen are or will be grown in small scale 300 

mL bubbling columns I the lab to obtain biomass and lipid productivity data. Furthermore, two strains 

have already being tested by a collaborating laboratory for several months in the past year at the scale of 

30 Gal in the greenhouse. Both strains performed well in the testbed cultures, thus serving as proof-of 

concept that strains identified in our screen can be scaled up. Further, in a separate project which is part of 

the NAABB consortium candidate strains identified in this laboratory will be grown in testbed facilities 

very soon to learn more about the strains identified so far. The results will then be used to assess the 

progress of this project. 







The development of the methodology can be highly beneficial in many of the studies. The generation of 

the data library would also be of use. Prediction should become the source of hypotheses for further 

studies. 


The projectmay not have adequately considered how to compare strains from the new collecting effort 

with strains that are already considered potential feedstock or known from commercial sources to 

accumulate lipid. I would be more comfortable with the relevance of the project if hypotheses about the 

habitat's type-to algal lipid production had been clearer. 


Yes relevant. 


Although the goal of the algal screening project is relevant to program goals, it is highly unlikely that it 

will contribute to meeting those goals. 


As this is another culture collection that will likely not be applied in raceways or large scale commercial 

algae cultivation, the relevance to the Biomass Program's mission is difficult to see, but culture 

collections that are screened are important from an academic sense. 

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Strain selection is critical. The selection process needs to be proven against realistic outdoor systems. In 

relevant outdoor sytems Are the strains selected by this screening any better than randomly selected 

strains? 


The ongoing screening activities are not isolated from other activities, but rather an integral part of the 

research on strains within the PI’s laboratory. The sampling and isolation activities for the strains to be 

tested have already been completed through another project. In response to one reviewer about specific 

habitat selection, yes, during collection small shallow habitats that are exposed to drastic temperature and 

irradiance changes were sampled with the expectation that this might result in potential candidates for 

later use in raceway ponds. Also, yes, candidate strains that passed the first level screen are being mapped 

back to the habitats they originated from. Some of the candidate strains identified through our previous 

nile red screen mapped back to shallow habitats that were exposed to drastic daily changes in temperature 

and irradiance. However, that work is not part of this project. Here strains that had been isolated from 

habitats previously and therefore already exist in the lab are being screened. In response to the reviewer’s 

comment on strain selection, yes this is being considered, but also not as part of this project. Again, in 

another accompanying project the candidate strains identified are being tested in small-scale laboratory 

culture against strains such as the marine 'production strain' Nannochloropsis salina and a freshwater 

'production strain' Chlorella strain used by companies. However, not enough data exist yet to answer the 

questions if our screening effort results in strains that will be better than randomly selected strains. 

Currently, strains are being transferred to testbed facilities to answer the questions on performance of our 

novel strains in small raceway ponds. 








assessments) 




end-use) 




Critical Success Factors are not really considered in the framework of how this project fits in to the 

overall program, nor are they cast as desired. They tend to center on the laboratory experimentation itself. 


Exploring growth conditions that will be economical at commercial scale for any promising targets is 

important. Nile red screening is useful to eliminate most new collections, whose maintenance will be 

expensive; after that, analytical lipid characterization may provide more useful information. 

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The only success factor listed on Slide 12 is that high biomass/lipid productivity strains are identified. 

The work plan does not show how this productivity is to be measured. Define further the methods for 

culturing/testing of promising strains. 


Critical success factors were identified, but there is no plan how these obvious factors will be overcome. 


Lipid content is the obvious factor, but there was no real correlation to the conditions one would find in a 

raceway. 


Proof that the screening system selects superior strains is a critical success factor that does not appear to 

be addressed. 


In contrast to the reviewer’s comment, that ‘critical success factors…tend to center on the laboratory 

experimentation itself’, the ongoing project is embedded in other collaborative algae biofuels activities 

such as the NAABB. The methodology to be developed is supposed to replace the Nile Red screen as an 

analytical method that is as fast the Nile Red analysis, but providing more information on the lipid 

composition at the same time. Although testing of novel strains is not part of the current project, it is done 

in this laboratory in other projects. In response to the request to describe the productivity of strains the 

following is provided: Biomass productivity is measured as accumulation of biomass as Ash Free Dry 

Weight (AFDW) per volume per time. Algal cultures of about 0.3 Liters are grown in bubbling columns 

provided with 1% CO2 in air and illuminated with daylight fluorescent lamps at an intensity of about 200 

µE m-2 s-1. Cultures are kept in a water bath to regulate the temperature. Growth curves are based on the 

AFDW and lipid extractions are performed according to Bigogno [Bigogno, C., Inna Khozin, G., Sammy, 

B., Avigad,V., Zvi, C., 2002. Phytochemistry. 60 (5), 497-503]. From the biomass accumulation over 

time and the total lipid content at certain times during growth, the lipid productivities are being 

calculated. The PI agrees with the reviewer’s assessment that the screening, which is performed in the 

laboratory, does not have strong correlations to potential large-scale outdoor conditions. However, 

previous work of the PI on a saline water production strain has shown that the laboratory performance 

tests allow prediction of the strains behavior outdoors. 

Proof that the screening system used will select superior strains is not available yet as work on the proofof-concept 

relies on ongoing experiments in collaborating testbed facilities and is not part of this project. 

Although the proof-of-concept research is not directly part of this project, the results of that work will be 

the 'critical success factor'. However, the proof-of-concept work is in a stage too early to make any 

conclusions or predictions at this time. 





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The analytical methodology is a fine example of positive technology transfer. Data collection from a 

variety of samples gives some insight into the natural variations in lipid profiles that may be encountered. 

It may serve as a platform for further studies on improvements. 


The project began very recently (September 2010). 


Partners are present. Plan for feedback of information to Brooklyn College is not described. 


No technology transfer partners were identified. No technology transfer (outside of publications) is likely. 


Why not work with UTEX or other established collections? Why reinvent the wheel? 


There are many strain screening efforts. Communication among the projects, particularly regarding 

validation of screening systems should be improved. 

Defintion of performance criteria necessary and sufficient to enable real-world cultivation is needed. 

Wti teh lare number of strains that have been sampeld and screened, a statisical measurement of 

biodiversity could be used to estimate the probablity of finding a wild type strain with necessary and 

sufficient traits throuigh continued bio-prospecting. 


For clarification, technology transfer has already occurred, because in the past several candidate strains 

were forwarded to collaborating laboratories for further testing. 

The new screening approach using the UHPLC-TOF-MS system and the resulting data will be shared and 

reported on for example at professional meetings such as the US DOE User Meeting in March 2011 in 

Walnut Creek, CA, USA and the 3rd Algae World Summit in May 2011 in San Diego, CA, USA. Testing 

of strains in larger facilities is a goal of the overall research effort of the PI regarding strain 

isolation/screening. Although not specifically mentioned previously and in contrast to one reviewer’s 

comment that ‘no technology transfer (outside of publications) is likely’, some candidate strains identified 

through the previous Nile Red screening process were already passed on to collaborating Air Force 

laboratories for further studies. The Air Force collaborators report back to the PI on the performance of 

the strains tested. The Material Transfer Agreements for strain transfer are already in place for Air Force 

facilities and are also now ready for the NAABB. Consequently, strains are already in the process to be 

transferred to NAABB testbed facilities. Possible NAABB testing sites are located in Texas, New 

Mexico, and/or Arizona. Results on testbed performance of new candidate strains will be reported back to 

this PI so that the isolation and screening efforts can be adjusted depending on the outcomes of the 

testing. 

Page 191 of 223









Overall, I like the potential outcomes. I think the Investigators should concentrate on delivering the 

methodology, and not be concerned with attempting to identify the "best" algae. 


Assessment of new microalgal strains for feedstock potential is reasonable, but is best done with more 

explicit hypothesis-driven selection of habitats. It will be important to do broad screening and quickly 

eliminate most strains collected to date with Nile Red staining before doing analytical chemistry to 

precisely define lipids. Growth vs lipid accumulation conditions for new strains with promising lipid 

accumulations should be defined as early as possible. 


See comments above. 


Since the presentation only focused on the strain screening project, this review only covers that section of 

the project. It appears that the screen straining project was primarily focused on capacity building for the 

college (mainly an equipment purchase). In that regard, it is successful. However, this project's likely 

impact on the DOE program will be minimal. 


As yet another culture collection and screening of lipids what is the point and the contribution to the 

existing body of collections? 


The project would benefit from validation of the screening process. Outdoor pilot production testing of 

several strains rank ordered by the screening process could be used to determine if screening performance 

correlates to real productivity. 


The main purpose of this grant was increasing of the research/teaching capacities at Brooklyn College of 

CUNY. However, this particular research project is embedded into other ongoing algae biofuels activities 

funded by the AFOSR and the US DOE. For example, the PI of this grant is member of the National 

Alliance for Algae Biofuels & Bioproducts. In response to the reviewer’s comments on ‘validation of the 

screen’ and the ‘minimal impact’ of the project, this PI would like to point out that it is a priority of this 

laboratory to transfer strains that passed the screening process either with Nile Red or with the new 

UHPLC-TOF-MS approach as quickly as possible to testbed facilities either within or outside the 

NAABB. For example, some of the first strains identified as potential feedstock producers for biofuels 

applications have already being tested successfully in the greenhouse at a scale of about 30 Gal in the past 

year. Therefore, the impact of this project may not be as minimal as expected by the reviewers. Also, 

more candidate strains are ready for testing at a larger scale and as the screening continues, it is expected 

that even more strains will be identified for further testing. Although the reviewer’s have negative 

predictions, only the actual cultivation of candidate strains in raceway ponds or photobioreactors at 

Page 192 of 223




testbed facilities will provide answers to the reviewer's comments. At this time, it is this PI's opinion that 

it is too early to dismiss the approach taken by this laboratory. 

Project: 7.9.1.4 Cold Spring Harbor Laboratory 

Title: Exploiting aquatic flowering plants (duckweed) as a source of bioenergy 

Presenter: Rob Martienssen 



Approach 3.83 2.11 6 

Progress 5.20 1.47 5 










This project appears to lack strong foundation, if the intent is to develop an alternative lipid source for 

certain situations. The highest priority as demonstrated in the presentation is obviously the genomics 

work. The foundation questions such as: what are the growth areas for this oil precursor crop, and is this 

significant; what are the lipid levels currently, and how does this compare to various feedstock 

alternatives; what is the composition of the lipids; should this feedstock be considered for starch and 

subsequent conversion rather than lipids; what is the constituent composition of duckweed; has anyone 

done some preliminary work in this area before? It seems that no one has presented the case for doing this 

work, let alone jump to the step of genomic sequencing and modification. 


This project is a sequencing /transcriptomics/stable transformation system project on duckweed with the 

aim of converting starch production in the turions to oil storage. Lemna is already used commercially in 

animal feeds and bioremediation of waster waters in some areas of the world, making duckweeds 

potential biofuels' targets if GMOs will be acceptable in commercial scale and if the PIs hope to transform 

duckweed to produce oil in turion proves possible. Some milestones did not appear to be time-defined in 

ways that lead to practical biofuels' advances for lipid production. For example, what will project 

personnel do if duckweed can not be stably transformed? Turion production (techniques other than cold) 

and capability to produce turions at commercially valuable time periods were not described. Is there 

additional background literature on this? The well-described dispersal of duckweeds by birds needs some 

practical consideration by the investigators. 


Develop aquatic flowering plants (Lemnaceae) as alternatives to algae for biofuel feedstocks —Determine 

the genome sequence of several duckweed species, and screen for lipid content. —Develop genetic tools 

to manipulate oil content in fast growing duckweed species 

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For lemna to be a useful biofuel crop, it must have high biomass and/or oil productivity under mass 

cultivation conditions. The presenter stated that his projections of high biomass productivity stem from 

commercial sources. These may not be reliable. For example, low productivity (~4 g/m2-d) was measured 

by Ryer in the 1970s. A literature review on lemna productivity should be a high priority. 

What is the potential to genetically manipulate lemna to have high lipid productivity if they are a 

naturally low productivity organism? Are the lemna production facility designs technically and 

economically feasible. This is presumably not the expertise of the presenter, who is a biologist, but since 

the stated goal of the project is to create a biofuel crop, these techno-economic matters must be addressed. 

The statement "Lemna spp. are already in widespread commercial use for animal feed and bioremediation 

of waste water" is an exaggeration. There may be a handful of sites in existence. Lemna Corp. was not 

very successful in commercializing lemna wastewater treatment in the 1990s. 


The objective is to increase oil content of duckweed. The approach is genetic: genome sequencing of 

lemna, followed by development of transformation and expression system in duckweed. Although the 

science is sound, the presenter did not convey any understanding of the larger picture (use and value of 

duckweed production systems). This lack of understanding unnecessarily limits the value of this research. 


Independent varification of growth rate claims should be made before research work is initiated. 


We apologize for not giving a longer introduction to the considerable research already published 

concerning the enormous growth and yield potential of duckweed and other aquatic plants as biofuel 

feedstocks. The Ryan study is generally considered an outlier in these studies. Geographic distributions of 

each species were determined by Elias Landolt in amassing a live culture collection of 1300 isolates. The 

phytochemical composition of the Lemnaoideae, including carbohydrate, lipid, and protein content, has 

been studied since the early 20th century. Cheng and Stomp (2009) provide a good survey of potential 

commercialization avenues for duckweed aquaculture, including promising data on efficient starch 

production. Duckweed may constitute an excellent feedstock for ethanol biofuels, yet this does not 

diminish the value of investigating the potential for lipid production in the plant, as the demand for lipidbased 

biofuels remains. Cheng and Stomp (2009): 

http://www.mekarn.org/MSc_CTU/literature/cheng%20et%20al%202009.pdf We recommend the 

following white paper as a source of reviews: 

http://lemna.rutgers.edu/Michael_duckweed_JGIDOE_2008_final.pdf This white paper forms the basis 

for the S.polyrhiza genome sequencing program at the DOE JGI genome sequencing center. As described 

during our presentation, our program will build on this reference genome sequence, first by sequencing 

several other genera that have superior potential as biofuels (eg. Oil content, geographical range, genetic 

properties). We have begun with Lemna gibba, described in the presentation. With respect to progress in 

our research proposal, preliminary analysis of TAG content in several accessions of S. polyrhiza was 

presented in additional slides at the end of the presentation. However, new technology developed over the 

last decade has made genomics a powerful alternative to classical physiological analysis of lipid and other 

feedstock composition. As we described in the presentation, the gene catalog will form the foundation for 

a model of carbon paritioning that can be further tested in a more diected way. By analogy, the genome of 

microalgae and diatoms has proven to be an essential component of biofuel strategies seeking to increase 

lipid content. We learned at the review meeting that lipid content is the limiting factor in energy balance 

and so we have made it the focus of our project. We have already made substantial progress in stable 

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transformation as well as in manipulating growth conditions to induce flowers and turions, and our 

milestones and deliverables reflect these goals. Given the emphasis on basic biology, it is outside the 

scope of our project to investigate the ecology, dispersal, and regulatory issues surrounding aquatic plants 

as alternatives to micro- and macro-algae, though we agree that these are valid concerns for all three 

potential feedstocks. To address these concerns, we plan to interact with industrial partners around the 

world who are already developing duckweed for various purposes, including oil production, as well as 

ecologists, engineers and academics as described below. While it is clearly well outside the scope of our 

project to establish the pilot scale growing facilities required, these partners have well advanced, well 

funded programs in exactly this area, and we hope to establish a network of duckweed researchers 

worldwide through our web portal (see below). 







Adequate progress appears to be obtained on the plant biology aspect of the problem. No progress appears 

on understanding the nature of the lipids and starches and if the work is warranted. 


Application of Arabidopsis data on lipid pathways and control genes (pickle) is promising. The work to 

date on gene silencing by P19 to maintain stable transformation in that way seems fruitful. 


Have established a facile and efficient transformation protocol that does not rely on transformation of 

callus tissues and regeneration with high transient transformation efficiency (60%+) and significant stable 

transformation rate (33%) in ~4 weeks 


Progress has been made toward genetic manipulation. 


The project appears to be on track for achieving its stated objectives. 


A strong demonstration of biotechnology, but there are many unresolved questions about the biofuel 

potential of duckweed. 


We thank the reviewers for their very positive response to our substantial progress in the first few months 

of the project. 

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It is difficult to judge the true relevance. Certainly alternative, high lipid feedstocks can be considered a 

Platform goal. It is not clear whether this duckweed is a legitimate choice for a feedstock. It grows rapidly 

in some places, is easier to harvest than algae probably, and there are lipids and starches produced. But 

there was no discussion as to why choose duckweed? Some catalog of the reasons for this organism as 

opposed to all others should be a basis, but is not presented. What previous work has been done? Is there 

some primitive technical analysis and even LCA that justifies this project? 


There is a large degree of transcriptomics/genomics work here that appears to represent excellent basic 

science; its payoff for biofuels' development is hard to evaluate because applications are beyond the scope 

of the project. 


Unclear. Although potentially good science, how could this approach translate into oil accumulation in 

Lemna? This is not discussed. 


Relevance has not been established. See other comments. 


Duckweed is an interesting model plant system but there is no objective evidence that duckweed 

production systems are viable for large scale production. In the absence of such evidence, this project is 

not relevant to program goals. 


If the duckweed productivity is as high as claimed, and if harvest and dewatering is easy, then the 

biomass should be a good feedstock even without high oil content. 


As we described in the presentation, duckweed is the smallest flowering plant, with one of the highest 

increases in biomass among known species. Given the very high level of investment in microalgae to 

date, both in the private and public sector, a reluctance to consider alternatives such as macroalgae and 

aquatic flowering plants is certainly understandable. However, the very high yields and ease of harvest 

are very well established, and our project therefore focuses on the promise of increasing lipid content. 





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assessments) 




end-use) 




This analysis is sorely needed for this project. The baseline estimation of 1) technical likelihood of 

success at creating a high lipid strain by genetic manipulation; and 2) given other attempts at creating 

high lipid organisms, what is the most likely level achievable with duckweed; and 3) given the best 

estimates, and coupling with growth rates and other feedstock generation features, what is the 

productivity and value as an endproduct? The question then can be addressed whether the project is worth 

doing under the most favorable outcomes. 


If turions can not be turned into oil storage structures, the project will not have met its goals. What would 

be the PI's default strategy for making a contribution to biofuels' development? 


Unclear. 


See comment on approach 


The project has defined and addressed critical success factors within its narrow focus of molecular 

biology. However, the project is unlikely to impact the viability to biomass feedstock supply and 

conversion. 


Success appears to be determined by the productivity of duckweed and potential use as biofuel feedstock. 


We have already accomplished two of the critical success factors - determining the gene catalog of 

duckweed, and developing an efficient transformation protocol, as we described in the presentation. 

While the typical lipid content of vegetative fronds varies from only 2-9%, concentrations of up to 26% 

have been observed in cultures augmented with phytohormones (Chang et al. 1978, Le Pabic 1980). 

Depletion of nitrogen or total minerals has been shown to induce turion formation (Czopek 1963, 

Henssen 1954), and indeed Cheng and Stomp (2009) found enhanced starch accumulation under a regime 

of 5-day post-growth transfer to water. We plan to profile this developmental shift to determine whether 

full turion induction is necessary for substantial accumulation of stored energy. 

Page 197 of 223










There is little doubt that publications will result from this work. The project is in line with genomic and 

biology type work. Unless the fundamental questions are addressed as to outcome estimation, it is not 

known how this will benefit the Biomass Program. 


The project is a sequencing and molecular biology project and publications in this area are good. 


There is a company in Florida (Petroalgae) that is already cultivating Lemna to large scale for biomass 

and fuels generation. In the context of this EERE effort, it would be an enormous plus for the PIs of this 

project to link with this company and to work jointly in this endeavor. Collaboration with Petroalgae 

would afford an applied dimension to this project, which is a mission of the EERE program. 


Several institutions involved. Commercial lemna technology firms were mentioned, but none are partners, 

which is a weakness. 


Technology transfer within the basic sciences is likely. However, it is unlikely that the Biomass program 

will benefit. 


Web portal to share data. 


We thank the reviewers for the excellent suggestion to partner with Petroalgae, which has already begun 

commercializing duckweed as an alternative to microalgae. The company raised significant capital 

through a recent IPO based on this technology. Our web portal is scheduled for release this summer and 

will provide an international hub for duckweed researchers world-wide. Our meeting at Cold Spring 

Harbor in 2009 was the first of its type for several decades, and brought together representatives from 3 

companies (Biolex, BASF and Lemnatec) as well as academics from Japan, Europe, China and the USA. 

The participants have all agreed to contribute to the design and population of the web portal and 

associated databases. 




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I would like to be happy with this project. Another high lipid biomass feedstock with highly desirable 

cultivation, growth and productivity would be very desirable. The basic analysis of the reasons for doing 

this work were lacking from the presentation. 


This project is likely to be successful in adding to our knowledge of duckweeds and lipid synthesis 

pathway controls (e.g., transcription factors). It is harder to evaluate its potential to contribute to 

commercial, sustainable biofuels' production. 


This is mostly a basic research effort. Given the EERE support of this project, PIs ought to bring out and 

emphasize applied aspects in their work. 




This is a basic science project that should be funded by NSF. It has no relevance to the DOE biomass 

program. 


It is not clear if oil production will make the duckweed a better feedstock than starch production would. 


We thank the reviewers for their generally favorable comments, insight and advice. 

Project: 7.9.2.1 Old Dominion University 

Title: Developing new alternative energy in Virginia: Bio-diesel algae 

Presenter: Patrick Hatcher 



Approach 5.29 1.16 7 

Progress 5.43 1.50 7 








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This is a "demonstration" project of sorts. The chemistry is not defined. The device apparently is. Now it 

is up to the Investigator to demonstrate the deliverable. I hope there are firm deliverables which are 

framed in yields and substantiated with appropriate techno-economic analyses and LCA analysis. Merely 

tossing off the costs of raw materials as manageable, or by-products as useable are not adequate for the 

evaluation of this process concept. 


The purpose of the research is to achieve one-step chemical processing of algae to biofuels. Conversion of 

Scenedesmus gave methylene-like components that can produce "algae-crude". Challenges include 

cleanly separating volatiles and maintaining catalyst. The major gap appears to be clear demonstration 

that this apparently energy intensive process is actually a potential source of sustainable energy. Further, 

purported concerns about intellectual property kept the PI from addressing a key question about N and P 

recovery and recycling during the discussion. 


•Establishing a commercially viable process for the conversion of algal biomass to biodiesel will go a 

long way towards widescale algal farming to meet alternative fuel requirements for the future The 

approach is to scale-up a ODU-patented process for a single-step conversion of algae to FAME biodiesel. 


Goal is to develop and optimize an algae to biodiesel pilot plant. It is not clear how much biomass is 

needed for the testing; what biomass will be used for testing; or, if from the ODU ponds, how it will be 

harvested. Little detail is given on the planned testing procedures. 


The project was focused on a one-step conversion process of algae to biofuel. The approach appeared 

sound but, due to time constraints, it was difficult to adequately evaluate this approach. 


This is basically a pyrolysis approach that will suffer if the feedstock quality (moisture, pigments, 

carbohydrate/protein ratios) cannot be controlled, which is going to be difficult. The TMAH cost (or 

supply) is a critical factor, as well, and was mentioned but not presented. 

The PI claimed an energy balance being positive but showed very little data. 


Specific goals for process performance and economics would be helpful. 






Page 200 of 223






The device has been obtained as in shakedown studies. The real work begins now. 


The project ends in May 2011 and was considered 65% complete at the report interval. The lack of 

presentation of an energy budget at this stage of this energy conversion project is somewhat concerning. 


ODU has successfully selected, designed, and constructed a pilot-scale “algaenator” built around the 

design of a horizontal wiped-film reactor. It is recognized that significant progress has been achieved 

prior to funding by this program. 


Tested "algenator" which didn't work or this was determined from a desk study--not clear. Switched 

reactors and site. 


The project has made progress in achieving its intermediate goals but major components of the project 

(testing, optimization, and evaluation of conversion unit and characterization of products) are not 

complete. Unfortunately, it is likely that investigators will rush to "complete" this work by the project 

deadline (May 2011). 


The pilot plant demonstration facility is quite good. No results past that were really given other than PID 

diagrams and statements of hope. At minimum a thermodynamic analysis could be been calculated and 

presented. 


Detailed pilot data is avaiable, but were not presented. 








This may or may not turn out to be relevant. DOE should be pushing for a true determination of the value 

or potential for this technology by project end. 


Energy conversion processes are important, but must be demonstrated to be efficient to be a contributor to 

platform goals. 

Page 201 of 223





Project fits the goals because the main activity has been to construct the pilot-scale reactor, called 

algaenator, and to eventually evaluate its performance over a wide range of operating conditions. This fits 

the goals of the DOE OBP Program by developing an efficient and cost-effective conversion strategy for 

algae to biofuels. 


Wet processing of algae to fuel would be valuable. 


The project contributes toward meeting the platform goals and objectives. 


This project might prove excellent at targeting soybeans and not microalgae. So it fits DOE's platform in 

that regard. 


The need for feedstock drying or residual water vaporization is likely to make the overall energy balance 

unfavorable. The economics project a value of $0.50/kg for the fertilzer byproduct. This seems very high 

for a low grade fertilizer. 2/3 of projected income is from this fertilizer. 









assessments) 




end-use) 




I do not believe these are defined well-enough for this project. There should be stricter evaluations than 

for a research project. 


The project must identify plans to evaluate energy sustainability of the processes described. 


Aspects of the harvesting technology to be developed by ODU is already successfully applied by many 

commercially viable microalgal biotech companies, serving the nutraceutical industry. 

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Technical and economic factors to be considered, but what are the quantitative goals? 


Critical factors were identified, but the viability of the project is unclear. 


This was not developed well and in particular, the keys to a successful energy balance were not presented. 


The economics and energy balance are properly identifed as critical factors. Byproduct value drives 

economics and must be realistic at full scale. 








Certainly at this stage, there is little detail sharing. I do not believe there will be many publications unless 

highly successful. 


The project's self-described inability to answer questions about recycling of nutrients from the system etc. 

may limit its technical transfer, in addition to some concern about the need to adress energy costs of the 

process. 


ODU ought to link with many of the other "production" efforts supported by the program so that there is 

integration of effort. 


Reactor company is a partner, but no other universities or professors seem to be involved. 


Technology transfer efforts and partners were identified. 


The pilot facility is quite good and clearly of use to partners. 


Publications, presentations, and commerical motivation ensure broad sharing. Critical reveiw of 

assumptions and projections is needed. 

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I am leary of projects which seem to have a mystical component, and for which the science is not 

established and shared and discussed. Yet I believe this work should be funded because just maybe there 

is something there. In this case I would set very stringent criteria and deliverables and stick to them. 


The presentation made it diffifult to evaluate the practicality of the work. 


This is a worthwhile effort seeking to advance a single-step conversion of algae to biodiesel using heat, 

methanol, and a catalyst from a proof-of-concept (laboratory) to a pilot-scale algae-to-biodiesel reactor 

that will allow for testing of process commercialization potential. Given the focal point of this effort, and 

the fact that the facilities are in place, the Program should expect significant outcomes from this work. 


Plan looks reasonable based on limited information provided, but some preliminary economic assessment 

is expected at this stage of development. 


The project is relevant to program goals, but the presentation did not adequately address all of my 

questions. 


A good pilot facility made with little relevance to microalgae but again this system could work very well 

for another feedstock wherein pretreatment conditions could be controlled (likely soybeans). 


The process appears to have energy balance and economic challenges driven by the need for low moisture 

feedstock. 



Title: Canada Algal Collaboration-PNNL 

Presenter: Jon Magnuson 



Approach 3.33 1.37 6 

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Progress 3.67 1.49 6 










I understand some of the technical Approach, but am baffled by the rest. I understand that the kelp may be 

another feedstock ultimately. We should understand the composition and if it varies with the time of year 

or locale or other variable, this should be studied as well. Preliminary data was presented showing high 

carbohydrate levels. The approach at this point should be to consider the compositional analysis and then 

choose one or more optimal methods for recovering the energy in these components. It may be for fine 

chemical use, or saccharification, or other process alternative. Why immediately jump to conversion to 

bio-oil through hydrothermal liquefaction? What is the basis for this choice? In addition, the logistics of 

ultimate product production should be a factor in the design of an approach. Where is the feedstock 

produced, and how to transport to point of conversion is important. 


This project considers the potential use of kelps from Canadian waters in biomass to biofuels ' conversion 

based upon mannitol and laminarian content. The PIs should consider the data on carbohydrate 

concentrations of kelps and nutrient effects on carbohydrate content of kelps exists in the literature-, for 

example from eastern Canada (New Brunswick, T. Chopin), France (studies at Roscoff), and 

Japan/China. It is of concern that data presented on kelp carbohydrate content were from a single site on 

each side of the Canadian coast (and even lacked characterization of water mass nutrients from that one 

site/coast). Unreplicated data are likely to contribute to poor sensitivity of the anlyses. 


Use joint US-Canada resources and expertise to characterize macroalgae with regard to composition and 

conversion to bio-oil. Develop process to convert Atlantic and Pacific coastal biomass to bio-oil/biofuels. 

This approach is a non-starter as the ecological consequences of making biofuels from marine coastal 

macroalgae has many environmentally and ecological pitfalls that make it untenable. 


The seasonal sampling of seaweed for compositional analysis seems like a topic that would have been 

covered extensively in past ecological literature. No literature is mentioned. In any case, apparently 

seasonal samples are needed for hydrothermal treatment. By national lab standards, the budget is modest 

for monthly sampling and thermal conversion. Does this presentation represent some work being 

supported separately by the Canadian government? Is there any preliminary economic analysis of the 

seaweed to fuel process? What is the expectation on cost and resource availability, even at a preliminary 

level? It would be good for this project and the one described by Guri Roesijadi to clearly show the level 

of collaboration/integration. Only the Acknowledgments slide suggests any cross-over. It seems like a 

reasonable project, but one that could be better represented by an integration of the two presentations. 

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The goals of the project are to characterize macroalgae with respect to seasonal composition and 

conversion to bio-oil. The approach for the seasonal composition portion of the project appears to be very 

limited (sampling only one site on each of the two coasts). Moreover, it appears that an extensive 

knowledge base about seasonal composition already exists. 


Algal biofuel process development is a monumental challenge with a low probability of success. Adding 

an additonal constaint of Northern latitudes is a bad idea. 


Review comment 1: These are very good questions. Currently DOE is concentrating on biofuel 

production processes, not fine chemicals. Hydrothermal liquefaction was chosen because freshly 

harvested kelp is about 10% solids so a conversion process that doesn’t require extensive energy 

consumption due to drying processes would seem to be a good starting point. Our expertise in this area 

(Doug Elliott, world expert in biomass liquefaction processes) made it a good choice for a project with a 

very modest budget (and level of effort) by leveraging existing projects and infrastructure/equipment. 

Logistics is a necessary aspect of the total process but was beyond the scope of this effort. Since there is 

very little work in the area of converting macroalgae to fuels and we had a modest budget, we sought to 

focus on assessing composition of one type of kelp and one conversion process. We felt this information 

would be valuable for beginning to assess the feasibility of macroalgae as a feedstock based on technical 

data. Review comment 2: I made a mental slip at the review in regards to Thierry Chopin. Dr. O’Leary 

(our Canadian collaborator) is a colleague of Dr. Chopin’s and discussed the sample collection with him 

at the outset of the project. Apparently, Dr. Chopin would have required funds to help in the collection 

and shipping etc., which is a reasonable request but this project is actually based on the two nations 

funding their own scientists’ efforts. I have not found a full compositional analysis of the polysaccharide 

(alginate, fucoidan and laminarin) and sugar/polyol (mannitol) composition of Saccharina latissima as a 

percentage of dry weight in the literature and would be pleased if the reviewers were able to provide a 

specific reference(s). We consider this analysis of relevance to biomass conversion to biofuels and 

bioproducts via thermochemical or biochemical conversion processes. It is important to understand the 

carbohydrate composition with regard to biochemical conversion processes. We did derive our extraction 

and fractionation procedures from work done by Rioux et al. (Carbohydrate Polymers 69 (2007) 530) so 

we are aware of literature in this area. Water column nutrient analyses and many other variables would be 

valuable to include in a larger project. For our admittedly modest project we were focusing on examining 

geographic and seasonal variability in carbohydrate composition since that scope fit within our budget 

and collaborative strengths. We believe this work is a valid addition to the sparse literature on 

carbohydrate composition of Saccharina spp. The scope of this project was not intended to create a model 

so the lack of “model sensitivity analysis” is a consequence of not having that scope (the suggestion of a 

preliminary model of the economics is an excellent idea and should be considered for future scope). 

Review comment 3: It would be useful if there was additional clarification and a literature ref. with regard 

to the reviewer comment that “making biofuels from…macroalgae has been shown to be untenable…”. 

New technologies will likely be required to support using macroalgae as a biofuels feedstock, as Dr. 

Roesijadi’s project on macroalgae resource analysis seems to suggest. This project is actually well linked 

to the other project on resource assessment, since Dr. Roesijadi is the lead on one of the two tasks in this 

project, as well as being the PI on the Macroalgae GIS analysis. Review comment 4: See comments above 

on literature Review comment 5: We respectfully disagree that “an extensive knowledge base about 

seasonal composition already exists”. See comments above on literature. Review comment 6: This project 

was funded as an outgrowth of the US-Canada Clean Energy Dialogue; hence it was conceived as a joint 

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US-Canada project. The location of Canada and the Washington coast in northern latitudes was part of the 

impetus for examining a cold water feedstock like the brown algae. 







I believe it will be difficult to bring the project to a real state of completion. The objectives could be 

altered to cover feedstock variations, logistics options, composition drivers and conversion options. If 

hydrothermal liquefaction were a justifiable conversion tactic, some preliminary test could be run, 

without attempting to create a new upgrading chemistry. This would be a nice project, easily completed in 

the timeframe desired. Some analytical work has been done, and only part of the collection process has 

been completed. Several conversion experiments are described but lack context or justification. 


This study is 20% complete and started in May 2010. The PIs should reconsider some aspects of the 

scientific design of the project and make sure that they have adequately uncovered and applied the 

published literature on kelp productivity and carbohydrate content. 


Developed procedures for extraction and quantitation of polysaccharide and sugar alcohol components of 

macroalgae. Monthly collections began in Sept 2010, and are continuing through Aug 2011 


See Approach comments. 


Seasonal composition data is complete, but the progress toward hydrothermal conversion seems very 

limited (only two reactors tests to date). 


Screening strains may have benefit in climates better suited for algae production 


Review comment 1: With the modest resources available, we concentrated on compositional analysis and 

a hydrothermal liquefaction process that has about a three decade track record of use for biomass 

conversion so this study is viewed as testing and optimization of a developed conversion technology on a 

new feedstock. Two tests have been run as reported in the review. Also, we have done some previous 

work on macroalgae (Macrocystis), which suggested that it would work (Elliott, D. C., L. J. Sealock, Jr. 

and R. S. Butner. 1988. "Product Analysis from Direct Liquefaction of Several High-Moisture Biomass 

Feedstocks." In: Pyrolysis Oils from Biomass: Producing, Analyzing and Upgrading - ACS Symposium 

Series 376, pp. 179-188). Converting the bio-oil to non-oxygenated biofuel by catalytic hydrotreating is a 

topic of significant interest in the biomass program and expertise at PNNL in this area is being leveraged. 

Review comment 2: The 20% completion is based on the Project Management Plan on file, in which 

projects have an end date of FY13 (as shown under the “Timeline” heading on the quad chart slide). This 

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would involve a more extensive work plan with bioconversion processes as well. Our progress has been 

good, under the less than optimal conditions that we have received none of our FY11 funding and 

subsequently were forced to trim scope, delay activities and reapportion remaining FY10 resources as a 

consequence. We have focused our resources on collection of the kelp samples for later compositional 

analysis since this is the time sensitive aspect of the proposal. Even though only two hydrothermal 

liquefaction tests have been run, there was demonstrable improvement from run 1 to run 2 on solids 

loading and conversion to bio-oil. Review comment 3: no reply needed Review comment 4: no reply 

needed Review comment 5: see reply to comment 2 Review comment 6: The “screening strains” 

comment seems to be misdirected, since we have no screening component in our project. 







The relevance discussion is lacking. What are the boundary conditions for kelp as a feedstock for biomass 

conversion? Without the knowledge of kelp yield and geographical availability, it is not known how 

valuable the project is. The choice to overlook optimization of the feedstock composition with respect to 

end bioproduct also limits understanding of the relevance. 


Kelp biomass conversion is a potential source of biofuel by biomass conversion , but project needs to 

demonstrate the economic feasibility and sustainability of such use relative to sustainable harvesting 

potential of wild crop or costs of aquaculture, including site selection aspects relevant to other uses of the 

potential growing area (navigation, other fisheries). 


Project may be relevant to the objectives of the Algae Biomass Program, but it will have catastrophic 

ecological consequences if implemented in a scale required for the generation of biofuels. 




The project goals are relevant to Program Plan goals. 


PNNL and previous microalgea analysis conclude that cold northern latituides are not a promising 

location. 


Review comment 1: Utilizing the guidance of the review template, we discussed where the project fit 

within the Biomass Program Multi-Year Program Plan and how it utilized relevant resources from 

Canada to leverage US DOE investment. Resource assessment with regard to geographically dependent 

yield data was not part of the scope of this project although it is an excellent suggestion. With respect to 

the reviewer comment: “The choice to overlook optimization of the feedstock composition with respect to 

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end bioproduct also limits understanding of the relevance”, the project is not proposing genetic 

engineering of Saccharina latissima although this fundamental suggestion would be of interest to other 

agencies. Our “end bioproduct” is a drop-in biofuel that would be derived via hydrothermal liquefaction 

and hydrotreating to make a renewable non-oxygenated liquid transportation fuel. Review comment 2: 

Site selection, economic analysis, aquaculture design and implementation are very worthy endeavors. 

However, such an expansive analysis was not within the scope of this project. I agree that this would be 

challenging and many factors need to be considered. Generating some (but certainly not all) of the 

technical data upon which to base decisions about the feasibility of macroalgae harvesting and/or 

aquaculture and subsequent conversion to biofuels and other bioproducts is one of the goals of this 

project. Review comment 3: Review comment 4: Review comment 5: Thank-you Review comment 6: 








assessments) 




end-use) 




A more detailed assessment of the critical success factors are highly warranted. To justify the project as 

beneficial to the coastal biofuels production where most of the world's population and demand exists is 

woefully inadequate as justification. It could be that the nature of the feedstock will require comminution, 

solar drying, and transport as low moisture tablets to specific, select coastal conversion centers where the 

polysaccharides are recovered for conversion to plastics in order to make true techno-economic sense. 

This sort of analysis is highly desirable, and has its own set of critical success factors. 


The scientific data input and market/regulatory environment for siting are not well considered and plans 

to make adjustments to this problem were not evident in the presentation. 


PIs ought to perform an ecological and environmental impact analysis to properly assess feasibility. 




Critical success factors were identified, but there was relatively little explicit discussion about the main 

challenges in kelp conversion and how they would be overcome. 

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The climate is a critical factor to success that this project cannot address. 


Review comment 1: We agree that economic analyses are highly desirable. This project had neither scope 

nor budget for such analyses initially, however, once technical feasibility is demonstrated and we have 

data a preliminary techno-economic analysis could be built. This project is more closely aligned with 

points ii (Feedstock Supply R&D) and iii (Downstream Refining R&D) in the review criteria. Therefore, 

we emphasized our critical success factors in light of those criteria, namely “biomass characteristics” 

from point ii and “conversion” from point iii. Review comment 2: These are very important and critical 

factors to consider in the context of an investigation of the broad issues surrounding the potential use of 

macroalgae as a feedstock. Unfortunately, this was not within the scope of our effort. Review comment 3: 

See responses to comments 1 and 2 Review comment 4: Review comment 5: We apologize to the 

reviewers, in the 10 minutes allotted for the presentation to cover the 12 mandatory slides in the template 

the level of detail had to be controlled within the presentation. The “challenge/solution” format was our 

attempt to address this in a few seconds. The solutions were presented in modest detail in the preceding 

“technical progress” slides. Review comment 6: We agree that the climate is a critical success factor for 

the growth of biomass feedstocks for conversion to biofuels. Brown algae (phaeophytes) grow best in 

cooler climates/waters and that was one of the reasons we chose to investigate them in the light of this 

US-Canadian project arising from the US-Canada Clean Energy Dialogue and the International program 

at DOE. 







I believe there could be potential here for knowledge generation that would contribute to the biomass 

program as a whole. It is not being addressed. 


The current work may not have a high potential for tech transfer. 


This may be irrelevant for macroalgae, pending the environmental impact analysis. Alternatively, know 

how from this effort may be applied to cultivated macroalgae or microalgae, projects that are currently 

supported by the Program. 


The Canadian partners are collecting Atlantic kelp; what else? What are NREL and SNL doing? 


Discussion of technology transfer was very limited. 


Broad sharing through publications and presentation. 

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General Comment by PI: This project was proposed and funded under the auspices of the DOE 

international program to encourage collaboration between US and Canadian scientists in areas of mutual 

interest under the US-Canada Clean Energy Dialogue. Review comment 1: We are generating knowledge 

about a biomass feedstock (kelp) common to both countries that has not been well studied in that regard, 

i.e., as a feedstock. Review comment 2: Review comment 3: Review comment 4: Dr. O’Leary and staff in 

Canada are collecting and shipping samples of kelp to the US for compositional analysis and conversion 

experiments. Dr. O’Leary is a source of considerable expertise in the area of macroalgae. Our Canadian 

colleagues are also providing expertise in DNA extraction from kelp species, primer selection and PCR 

amplification for genus and/or species confirmation by sequencing. This has been very helpful (efficient) 

since developing extraction procedures for new organisms is often rather time consuming. They are also 

performing these analyses on our Pacific coast samples. Unfortunately, there was limited time to go into 

details on everything we are doing. NREL and SNL are performing projects of mutual interest to US 

(DOE) and Canada (NRC) on microalgae. They were specifically scoped to be non-overlapping. 

However, we communicate monthly since all of the US national labs are working with the same Canadian 

partnering organization and it has been a very fruitful and satisfying collaboration in that regard. Review 

comment 5: Review comment 6: 






Some true justification studies would increase the value of this work significantly. The selection of 

conversion to bio-oil is not justified. If one were considering floating hydrothermal conversion plants, 

why? Should solar collection be used to create a new conversion process that might yield products with 

less oxygen content? So what is the basis for the work? More could be squeezed out of this. 


This project suffers from a weak design for new data collection and inadequate review of background 

literature. 


There are good elements in this work, pertaining to the development of technologies for the extraction of 

product from biomass. I recommend the PIs focus their efforts in this area, and please properly assess the 

impact of harvesting the oceans' algae for biomass and biofuels. 




The seasonal composition portion of the project may be of very limited value because of its sampling 

protocol. It was difficult to evaluate how aggressively the hydrothermal conversion processing portion of 

the project was being pursued. 

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Resources could be better allocated by removing geographical constraints and focusing on regions with a 

scientific basis for having a higher probablity of success. 


Review comment 1: The basis of the work in this modest effort was to collect some of the relevant data 

on S. latissima composition and one proven method of conversion that could work with wet feedstocks 

and utilize all the biomass (see comment 1 under section 2). Review comment 2: See comment 2 under 

section 1. Review comment 3: Thank you for your comments. Further resource assessment and an 

environmental impact study would be worthy additions to the research portfolio at DOE. Review 

comment 4: Review comment 5: Different variables of interest will be chosen by different investigators 

within the constraints of their program. Given that we needed to work with Canadian colleagues on the 

Atlantic coast as advocated by the Canadian Funding agency and we are located on the Pacific coast, this 

geographical variable was chosen. The other variable was the amount of different polysaccharides over 

the course of the annual growth cycle. This is of great relevance to potential biochemical conversion 

processes where it would be desirable to have the highest concentration (wt/wt total kelp biomass) of the 

more easily hydrolyzed and bio-converted laminarin (non-crystalline glucose polymer) and mannitol 

(directly utilizable sugar alcohol). The percentage of alginate and fucoidan would be relevant to both 

thermochemical and biochemical conversion processes with respect to solids loading possible with these 

highly viscous hydro-gel forming acidic polysaccharides. Pre-processing to address this issue was 

performed under the hydrothermal liquefaction task enabling solids loading of 22% after just two tests. 

The compositional analysis study was designed to collect data of relevance to both biochemical and 

thermochemical conversion processes to be of maximum relevance. The research on the hydrothermal 

conversion process represents approximately 20% (please check this number) of our effort to date. 

Review comment 6: 

Project: 9.6.6.2 Sandia National Labs 

Title: Modeling and Visualizing Algae Biofuel Production Potential in Canada 

Presenter: Howard Passell 



Approach 4.00 0.82 6 

Progress 4.50 0.76 6 










This is seemingly a viable approach. What could not be presented are the list of assumptions that are 

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behind this end-product. It should be a required output that these are reviewed with a panel of experts, 

and either accepted as logical or whetehr some several modifications should be made to increase the value 

of the model. I believe at this point, a techno-economic evaluation and LCA should be applied to the 

conclusions at this stage. If there is no hope, no further polishing of the model is required. 


This is a modelling project with sliders for average solar irradiance, environment specific parameters etc. 

It examines co-location tradeoffs (e.g., biofuels' facilities and power plants etc.). A major challenge for 

this project is to apply the theoretical data to real future production, which the PIs appear to recognize. 

Among the weaknesses of the model is one Redfield ratio for all algae (different taxonomic groups have 

different Redfield ratios), the theoretical productivity levels used, and co-location challenges. The project 

personnel may want to review published literature in the areas to improve the models. 


•To build a prototype computer simulation model that will: –allow users to evaluate and visualize algae 

cultivation and biodiesel production capabilities at multiple sites across Canada –allow users to simulate 

tradeoffs associated with different resource co-location strategies (wastewater, waste CO2, waste heat) – 

lay groundwork for larger scale algae biodiesel productivity modeling 


See comments under Relevance. 


The goal was to make a simulation model for multiple sites across Canada. The approach is technically 

sound but it was not clear to me whether this modelling exercise was an isolated effort or would 

contribute to other modelling efforts. 


The modeling should be used to underscore the fundemetnal disadvantges of northern locations. 








It is difficult to judge progress. While the model may be nearly complete, what are the outputs and how 

do they match with reason or reality? Also what were the inputs? Finally, what is the resulting case that 

the model delivers? Maybe a 1-day workshop/conference to cover these should be arranged before the 

project close, and certainly before further work is done. 


This project is 75% complete and will end in September 2011. The project will produce a theoretical 

product. 

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A model was designed that calculates theoretical algae biodiesel production capabilities in northern 

latitudes given actual municipal effluent volumes (with nutrient concentrations), recycled CO2, land 

availability, and climate/solar insolation. HOWEVER, missing appears to be the solar insolation as a 

parameter that defines productivity. 




The project seems to have achieved its stated milestones and is on track for completion. 


The model appears to give expected results. 








This could be highly relevant as a decision-generator if it has the acceptable assumptions and inputs. In 

this way it does help the BMP. 


The project's relevance would increase if better input data from the literature on productivity were added. 


Somewhat marginal. Not knowing the strains that would need to be cultivated undermines the effort. 


In January 2009, the British Columbia Innovation Council published a major report on the prospects for 

algae biofuel production in Canada. Their conclusion matched that published by others even earlier: the 

Canadian climate is not suitable for affordable phototrophic algae biofuel production using current 

technology at 7 g/m2-d annual average biomass productivity (nor reasonable projections of improved 

technologies 14 g/m2-d). The SNL presentation shows a figure on Slide 10 with 5.2 kg/m2-yr as a high 

projected productivity (in southern Ontario). This is equivalent to 14 g/m2-d, same as the previous work. 

The SNL team would need to acknowledge the previous work and give some reasons why the new effort 

would come to different conclusions. 


The project is relevant to program goals. 


The model can be relevant by advancing other modeling efforts. 

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The results of the model should be evidence of the futility of putting geographical constraints on algal 

biofuel technology development. 









assessments) 




end-use) 




This tool is based on some critical factors, and could be used to generate the critical decision on algae-tobiodiesel 

in northern climates. As such defining success for this project is not adequately done. Future 

model tuning is posed already without measuring the value of the model in its first generation. How to 

gain acceptance is also not part of the success factors, but clearly should be. 


Sites for co-location may need additional inspection relative to market and climate factors. An expert 

review of input data for the model relative to the productivity values used in the model would contribute 

to the value of the work. 


The PIs, to their credit, recognized that a challenge in their project is applying theoretical data to real 

future production. 




Although one of the critical success factors was whether the model would aid Canadian partners in site 

selection, there seemed to be no plan to address this obvious obstacle. 


The critical success factor is the use of the model to make good policy decisions. 

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The potential is there if brought to fruition. 


Checking the model would lead to more tech transfer (i.e check the algal physiology literature, check 

physical sites for co-location). 


Transfer of know-how gained in this effort should be straightforward, and the PIs are commented for 

planning to publish their results in the peer-reviewed literature. 




The main technology transfer effort seems to be toward adapting the model to other countries/regions. 


Publicatoins and presentations ensure good tech transfer. 







We may already know that algae is not viable as a feedstock for diesel in these northern climates. This 

tool should be used as is to bring this question to an answer. If we have to have municipal treatment 

plants available to provide nutrients, this certainly provides a boundary condition for biodiesel production 

possibility. If the question was answered, no more work on the model is warranted, and alternative 

solutions may be studied by scientists in both countries without polishing this work further. 


This project may have taken too theoretical an approach. 

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This project entails a useful background analysis that, once performed, should be available to the field for 

a long time to come. 




This seemed to be a reasonable modelling effort. Its impact (beyond model development) would be more 

significant if there was more interaction with potential industrial partners. 


Results of previous modeling clearly show that Northern lattitudes are very unlikely to be good locations 

for algal biofuel production. The goal should be re-directed to produce a good model that is not specific to 

a geography. 


Project: 9.6.6.1 NREL 

Title: US-Canada Algal Biofuels Partnership 

Presenter: Philip Pienkos 



Approach 3.40 1.36 5 

Progress 3.80 1.17 5 










The approach is open to question in that is not clear why the US labs, especially NREL, are doing what 

they are. Why should NREL generate a culture collection of 100 strains from Canadian waters? What are 

the Canadians doing? What is the pertinence or significance of the experimental approach outlined in the 

presentation? 


This project is a microalgal strain discovery and culture collection delivery grant for strains that grow 

well at northern (cold) latitudes. The biofuels' industry could be seasonable in Canada, to benefit from 

long photoperiod and suitable temperature in late spring-early fall; however, the presentation did not 

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strongly demonstrate the advantages of the new strains isolated to the latitude. Six different batches of 

128 water samples were assayed with robust growth in flue gas enriched, well-buffered media. Over 40 

unique strains have been identified to date, mostly green algae. The project developed a process to deliver 

simulated flue gas. They hope to bring Univ. of Minnesota personnel in during last part of project to assist 

with studies of wastewater nutrient supply and mixotrophic algal growth. 


Project Goals: •NREL: Bioprospecting for microalgal strains with suitable characteristics for cultivation 

in northern latitudes •SNL: Site analysis and production modeling for cultivation of microalgal strains in 

selected regions of Canada and northern US •PNNL: Evaluation of macroalgae species common to the US 

and Canada for biofuel production 


The stated objective is strain collection and screening for lipid producing strains and characterization of 

lipids (focusing on strains collected in Canada). However, in light of the repeated failure of previous algal 

screening efforts in yielding strains that successfully compete in full-scale outdoor raceways, there was no 

justification regarding why this particular effort would be more successful. 


Macroalgae for biofuel feedstock has a low probablity of success in any geography. Constraints to 

northern lattitiudes lack scientific basis. 


1.A. The overarching goal for the microalgal aspect of the collaboration to deploy algal cultivation and 

processing for production of biofuels. This is a long term goal and this small project is designed to 

provide the basis for more extensive and expensive work in the future. It can reasonably be asked why the 

US would be interested in algal cultivation in northern latitudes with better locations available in the 

south, but Canada does not have that geographical advantage. However, successful deployment of algal 

cultivation in Canada will mean that much more US geography can come into play, allowing for more 

distributed production and greater volumes of fuels produced. The time constraints for this presentation 

did not allow me to provide a better description of the Canadian partner’s role. They have identified the 

potential sites for deployment and collected water samples from those areas. They will share in the 

responsibility for evaluation of the strains and further development of those strains with best production 

characteristics. They are also responsible for development of the cultivation process that can best be 

deployed in northern locations. 1.B. The point about the seasonality of cultivation and thus the biological 

advantages of local isolates is well taken, though strains that are better adapted to lower average 

temperatures could be expected to maintain productivity throughout a longer growing period than isolates 

from warmer climates. 1C. The reviewer presents the dilemma for all algal bioprospecting efforts. It is 

impossible to evaluate a priori the impact of competing strains in large scale cultivation systems that have 

not yet been built, but it isn’t practical to build those systems without some expectation of the strains to 

be deployed. The decision to isolate strains from likely sites of cultivation is expected to mitigate the 

competition problem somewhat. Further mitigation of this problem can be achieved through the use of 

PBRs either for cultivation or for production of robust and high cell density inocula. The successful 

cultivation of a handful of production strains by Seambiotics is a case study in which seasonal variations 

in culture conditions can be managed by proper strain selection and rapid turnover of the ponds can be 

carried out through the availability of a controlled inoculum system indicates that this problem can be 

solved. While the open pond cultures at Seambiotics may not be axenic or even unialgal, they consist 

primarily of the production strains and they are grown in a consistent, reproducible manner. 1.D. 

Successful deployment is an economic issue not a scientific one. While the success with any 

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photosynthetic algal process for biofuel production may face a number of significant hurdles, the 

scientific basis for success and, in fact, the technical feasibility has already been demonstrated. As noted 

in my presentation, the added constraints for successful deployment in northern latitudes, cannot be 

ignored. However, this question is not simply one of probability of success, but must also take strategic 

value into account. 







Progress has slipped behind the original workplan. 


The project is 70% complete and ends in June, 2012. The process of strain discovery and transfer to a 

Canadian culture collection is nearly complete. Some aspects of nutrient supply are under investigation. 


Strategy for water sampling developed with NRC. 128 water samples received from Canada in six 

separate batches. Strain isolation proceeding well. Reviewer is making a recommendation for the 

deposition of useful strains in recognized algal libraries (e.g. UTEX). 


Given the funding level and project period, progress to date (40 strains added to the collection) is very 

limited. 


More progress could have been made through literature surveys and consultation with experts. 


2.A. Deposition of strains into a recognized algal culture collections is a good idea. I will share that with 

the NRC. 2.B. The funding level is insufficient to support a single technician; the strain isolation progress 

is proceeding at a pace commensurate with resources. Much of the early work was focused on the siting 

analysis for water sampling and the sampling procedures, the analysis of flue gas, and the development of 

a system to supply simulated flue gas in a safe manner. This latter process may seem straightforward, but 

was actually more complicated than expected. 2.C. It would be more helpful if you could provide specific 

examples of publically available information that could accelerate our progress. 





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I have difficulty following the relevance. Can we not use US samples from northern waters? Will algae 

culture in northern waters make any economic sense? How does flue gas factor in the relevance? 


The relevance of the strain discovery is unclear; more nutrient demand and lipid production 

characterization will be needed, based upon information presented. What is the sustainability (buffering 

of flue gas supply of CO2 etc.)? 


The work addresses goals in the area of Feedstock Supply Ft-B. Sustainable Production: By utilizing 

information generated by resource assessment (SNL) culture conditions can be established to match 

optimal deployment area. Ft-C. Feedstock Genetics and Development: Genetic diversity in algal culture 

collection will provide strains with potential for commercialization of algal biofuel production in vast 

area of North America beyond the southeastern US and gulf states. 


Although project goals are relevant to biomass program goals, there was no plausible explanation why 

this project would be successful in producing algal strains that would thrive in full scale production 

systems. 


The work is very unlikely to lead to advancement in biofuel technology. 


3.A. We believe that Canadian water samples have the potential to provide strains better suited for local 

deployment. Although, this may only provide a slight improvement in our overall probability of success, 

the water sampling was a task for the NRC. Hence there was no advantage to choosing US water samples. 

At the moment, even optimistic estimates for algal biofuel production costs without benefit of a high 

value co-product indicate that this approach cannot compete with petroleum. As I noted in my 

presentation, productivity in northern latitudes will make it even harder to compete on a straight biofuel 

production basis. The challenge for all will be to identify value-added co-products that are scalable with 

biofuels. Meeting that challenge is even greater for areas that are less than ideal for maximum 

productivity. There is a great deal of information in the literature regarding the toxicity of high 

concentrations of CO2 as well as the contaminating gases found in flue gas. Most of this work deals with 

enrichment cultures or small bench scale cultures grown from dilute inocula. Careful analysis of the body 

of work suggests that the problem was not the toxicity of the gas components, but rather, the acidic nature 

of CO2, NOx and SOx, but that is an unproven hypothesis. Therefore, to be certain that the strains 

isolated would be able to thrive with flue gas as a CO2 source, we planned our enrichments to isolate only 

strains that were robust enough to survive the challenge of simulated flue gas. As I noted, the enrichment 

cultures in well buffered media provided robust growth. This prompted us to modify our media 

formulation and the results support the hypothesis that the problem with flue gas is more a pH issue than a 

toxicity issue. 3.B. I agree that strain characterization is essential, and that has begun at NRC with the 

transfer of our current culture collection. The buffering was necessary for small scale enrichments and 

culture growth because pH control was not practical for the number of cultures being grown. But 

extensive buffering is not necessary for larger scale cultures. The pH shifts caused by CO2 dissolution 

can be used to control CO2 feeding using pH probes to control gas addition valves. This process is used 

to great success by our NRC colleagues. 3.C. See response 1.C. 3.D. The DOE algal biofuels roadmap 

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calls out the need for additional diversity in algal strains. The strains to be isolated in this project will add 

incrementally to that diversity. I am forced to agree that the scope of this project is currently too small to 

lead to any major advance, but the development of the strains that arise from this work whether by us or 

by other groups could lead to the identification of a robust production strain or to novel genes or 

pathways that could be exploited for production of biofuels or other industrial targets. 








assessments) 




end-use) 




There is no significant discussion of CSFs, only a general statement about commercialization of algal 

biofuels. 


The proposed collaboration with wastewater experts at Univ. MN seems likely to improve final product. 


Critical success factors for commercialization of algal biofuels span the value chain and include 

biological productivity, cultivation, harvest, extraction and conversion to fuel. This project is currently 

focused on biological productivity, namely growth and lipid content. 


Critical success factors were identified but there seemed to be no plan how the most predictable obstacle 

(failure of strains to thrive in full-scale systems) would be overcome. 


Climate is a critical success factor that cannot be addressed by this project. 


4.A. Time constraints did not allow me to develop critical success factors further. 4.B. The entire 

partnership is in agreement that UM should be included in the collaboration. We are looking for a funding 

mechanism to add them to the team. 4C. This obstacle was addressed in Response 1.C. but I believe that 

its predictability is overstated, and the growing number of organizations scaling up cultivation of specific 

strains supports that belief. 

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The program has not been defined adequately in ultimate objectives to suggest whether there will be tech 

transfer beneficial to the OBP efforts. 


The culture collection transfer to Canada is a solid achievement but its relevance to biofuels' development 

may require additional demonstration. 


This is primarily with Canada. PIs are urged to deposit strains in a recognized algal library in the US (e.g. 

UTEX) and overseas. 


The potential for significant technology transfer is very limited. 


It does not appear that the investigators did a thorough lieterature review. 


5.A. The mechanism for funding this project in very small increments and the short time allotted to the 

presentation led to a decision to focus on short term goals. Potential tech transfer opportunities are most 

like to arise from the identification of superior strains and their subsequent evaluation in productionrelevant 

conditions. 5.B. Demonstration of the value of the new isolates is certainly necessary for any 

measure of relevance. Whether that takes place within the partnership or is carried out by other 

investigators will depend on many factors such as future funding for follow on work or partnership 

discussions with other stakeholders. 5.C. See response 2.A. 5.D. See response 5.A. and 5.B. 






The investigators failed to explain the reasons for the work, nor describe the ultimate value to be gained. 

If there was a political desire to work with Canada, I would think that some better projects could have 

been developed, or the explanation performed better. 


Characterization of the biofuels' potential of the isolated strains would make the project's outcome clearer, 

especially in the context of a comparison of existing biofuels' feedstock strains. 

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Project overlaps other supported Algal Biomass projects where the emphasis is on "prospecting". The 

Aquatic Species Program already undertook this propecting approach some 20+ years ago. 


This appeared to be a very traditional approach to strain collection and screening (although focusing on 

strains collected in Canada). Given past failures by other such projects, this approach, there seems little 

reason to support this project. 


There are sound scientific reasons for this project to have a very low probablity of advancing biofuel 

technology. There is no identified sound scientific reason to pursue macroalgae biofuel production in 

northern climates. 


6.A. The time constraints and strict format for each presentation made it difficult to discuss the reasons 

for the work in any detail. The value of indigenous production strains for local deployment should have 

been made more explicit. 6.B. Characterization of the isolates was noted in the future work section. I 

agree that it would be useful to compare productivity with some benchmark strains, and we will include 

that in our future plan. 6.C. The trains isolated for the ASP were isolated from water samples primarily 

from the southwest and southeast US with some sampling done in Hawaii and the northwest. The most 

interesting of those strains were transferred to the University of Hawaii for curation and many if not most 

of those have been lost. If the hypothesis that indigenous strains can be more competitive in local 

cultivation systems is true, then it certainly follows that local sampling and strain isolation is a valid start 

to a new biofuels project. Beyond that, we are beginning to see local communities push back against scale 

up of non-indigenous strains (e.g. in Texas and Hawaii). The ASP demonstrated that prospecting could be 

successful, but did not end the need for prospecting. 6.D. Presumably the comment refers to failure to 

successfully deploy the strains isolated in past efforts. Again this goes back to the notion that only “weed” 

algae can be successfully cultivated outdoors. If that were true then only heterotrophic algae can be 

expected to succeed, because regular production of high lipid biomass cannot be accomplished without 

long term cultivation of a production strain, whether that strain is a natural isolate or an engineered strain. 

6.E. Nearly all worthwhile endeavors begin with a low probability of success. This project is a tiny 

element in a very large distributed endeavor. I would expect the scale of advance to match the resources 

available. 

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Reviewer Comments - EERE

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