Progress 06/01/15 to 01/31/16
Outputs
Target Audience:Target audience(s) described in the Project Initiation Report identified three potential market areas. First area included kraft pulp and dissolving pulp. Kraft pulp is derived from wood and is close to being pure cellulose. It is used primarily for paper and paperboard production. Kraft processing requires mechanical and chemical treatment of wood, which can result in malodorus emissions and generation of wastewater with high pollution potential. Dissolving pulp is made from pulpwood using sulfite or kraft process with acid prehydrolysis for removal of hemicelluloses resulting in cellulose content of around 90%. Dissolving pulp is used for making viscose (textiles) or derivatized cellulose such as cellulose triacetate (films) and methyl cellulose (thickener). Both kraft and dissolving pulp are global commodities produced in the millions of tons per year. Dissolving pulp usage globally in 2013 was 6.0 million tonnes. The average price of kraft pulp is $700/ton, and dissolving pulp, $1,000/ton. Since algal cellulose is mostly lignin free, algal cellulose is easier to process, more environmentally benign, and less costly to produce. However, because of the large quantities involved in kraft and dissolving pulp market as well as the existing infrastructure that supports these markets, we will not participate in either the kraft or dissolved pulp market. In an effort to identify target audience for more specialized application of cellulose for textile, we collaborated with Cellulose Science Incorporated (CSI), Madison WI. Since CSI had been in the cellulose business since 2007, as developer of technologies for utilizing celluloses and lignocelluloses, we thought that CSI might aid in identifying potential end-users of algal derived cellulose. Samples of dry extracted algal cellulose were provided to CSI for their use in contacting potential end-users. The effort did not yield any potential end-users. A sub market to large quantity kraft pulp markets, is a small quantity cellulose market for specialty papers. Wisconsin ranks first in volume of paper manufactured and in diversity of paper products produced. A list of paper products that could incorporate algal derived cellulose include coated paper, specialty paper including velvet, chromo and chipboard, synthetic paper used for labeling, and security water marked paper. At least 14 specialty paper suppliers have been identified as target audience in Wisconsin. The second market area identified for target audience was for microcrystalline cellulose (MCC). The market for MCC is fairly well established. The existing global market for MCC is around $500 million and is projected to increase to $940 million by 2020. Pharmaceutical industry is the largest market segment for MCC where it is used as drug excipient and other medical formulations. MCC is also widely used in food industry in products such as bakery goods, beverages, dairy products, dressings, and sauces and in cosmetics. MCC in the current market is processed exclusively from woody cellulose. MCC derived from algal cellulose would need to be grown in clean water with synthetic chemical nutrients. All aspects of cultivation and processing would have to meet strict FDA requirements, which inevitably would result in increased production cost. Given that production cost for MCC is less than $5.00/kg, the large margin would justify the added cost foe clean cultivation and processing. As an example, small quantity of MCC can be purchased on-line from suppliers. Sample pricing for non-food MCC is $33.90/kg from Avicel, and $24.95/kg from Chem Center. Pricing for food grade MCC from Loudwolf is $141/kg. There are a number of large companies in the pharmaceutical, food and cosmetic MCC market. They include Asahi Kasei, FMC, DFE Pharma, J. Rettenmaier & Sohne, Libraw Pharma, Juku Orchem, Sigachi, Avantor Performance Materials, Mingtai Chemical and Accent Microcell. Algae derived MCC market could be a supplement to woody biomass MCC market, but the end users would be in applications such as specialty paper and paper board products, bio-composites (with polymers), filter medium, gas sensor devices, ion exchange membranes, and energy storage devices. We have identified target audience for potential application of algal MCC in specialty paper and filter medium. The nanocellulose materials, cellulose nanocrystals (CNC) and cellulose nanofibrils (CNF), market is segmented by anticipated demand; large volume, small volume and low volume/novel applications. Large volume market includes application of nanocellulose materials in packaging coatings, hygiene and absorbents, automotive body and interiors, textile for clothing, and paper coatings. The small volume market includes application in wallboard facing, insulation, aerospace structures and interiors, and aerogels for oil and gas industry. Identified target audience in nanocellulose application include paper coatings and filter medium. To get a sense of market size, the anticipated US demand for large and small volume market is 5,966 and 480 thousand metric tonnes, respectively. Using the cost of producing nanocellulose materials at $4.4 to $11.0 per kg, the cost of production is $26.25 to $65.6 billion for large volume, and $2.11 to $5.28 billion for small volume nanocellulose materials market. During the period of this project, individuals and groups conducting "cutting-edge" research on nanocellulose materials reported to us that they have no sources of algal celluloses other than through our company. We provided unprocessed algal cellulose to researchers, but also conceived plans to distribute product through chemical distributors. We forged a partnership with another company located in Madison, WI that operates in a complementary sphere, namely serving customers by coping with nuisance algal growths. Based on this experience, we perceive that our expertise in harvesting and processing algal biomass, and our expertise in microscopic and molecular assessment of nuisance algae will prove widely valuable to private and public entities. Municipal wastewater treatment plant is the third area for target audience. Algae need water, nutrients, carbon dioxide and light to grow. All of these ingredients are available for free at municipal wastewater treatment plants. Integrating algal cultivation in wastewater treatment provides nutrient removal service while growing biomass. Co-locating algal production at a wastewater treatment plant obviates the need for a new source of water and makes use of existing infrastructure, i.e., tanks, piping, dewatering equipment, drying equipment, power, etc. Non-cellulose component of algae may be valorized through digestion to produce biogas to generate electricity and heat required for plant operation or further processed yield fertilizers or lipids for biofuel. Moreover, any process waste by-products will most likely be recyclable for treatment. Thus co-locating algae cultivation at a municipal wastewater treatment represents the lowest cost option for scaling-up production of cellulose products. Municipalities that own the treatment plants would be incentivized to adopt our algal cultivation technology because algae would enhance nutrient removal. The algal biomass resulting from cultivation could be sold and create a revenue stream. In all but the most land limited wastewater treatment plants, some degree of algae cultivation can be adopted at treatment plants. Where land is available at relatively low cost such as at wastewater treatment that serve small rural communities, our algal technology may function as a significant economic boost for the community. Our on-going collaboration with Madison Metropolitan Sewerage District will serve as an example for other wastewater treatment plants to consider and adopt our algal technology. Changes/Problems:Changes and problems encountered during the Phase I project centered on stock biomass cultivation, and inability to follow the planned scope of work. Biomass was cultured in carboys for 1-2 weeks prior to inoculation in the larger reactors. The long time between stock inoculation and it being ready for reactor inoculation limited the time for accomplishing the experiments and caused several experiments to be conducted later than desired in the growth season. The reason for the long stock culture preparation time was a result of several factors. First, overexposure to sunlight was a constant concern. Stock cultures were initially put into full sunlight before the first run, and within an hour, significant portions of the biomass had bleached out and died. This resulted in several delays during the stock culture preparation times. However, this observation suggests that Oedogonium is able to utilize lower light levels for optimal growth, and so culturing (in stock or in full bioreactor designs) do not need have high concerns for light availability, which may facilitate the use of this system in areas that are normally not thought of as having enough light for algal growth on large scales. The observation leads us to place density-neutral filters over each reactor in order to shade the biomass from full sun exposure, reducing the change of photo damage. The experiments also were modified to account for several discrepancies that came up during the project. The manipulation of mixing within the reactors was dependent upon air pumps and CO2 discharge system. The low cost air pumps could not pump at lower rates and maintain airflow through the lines to push air into the reactor; therefore, it was not possible to test the effects of high CO2 with low mixing in the system. All reactors received a high level of mixing at all times, and this was held constant throughout all experiments. Elevated nutrient feed to determine effects on biomass production was not performed due to inability to access and store high nutrient process streams. What opportunities for training and professional development has the project provided?This project employed both manual labor in the production process and highly educated personnel to perform laboratory processing to high quality standards, an experience leading us to believe that our process has the potential to employ both highly educated and poorly educated, disadvantaged population groups. This project provided on-the job training and professional development for project personnel; Mike Piotrowski, Izak Smith and Zach Muscavitch. Linda Graham PhD and James Graham PhD provided mentoring in the area of phycological science and applied phycology. Jun Yoshitani, an experienced wastewater engineer provided mentoring in the area of wastewater treatment system. The project personnel also gained experience in design, installation and operation of a field scale algal cultivation system (liquid volume of 6,200 L and a footprint of over 40 m-2). They learned how to install a pH control system using carbon dioxide, pH sensors, solenoid control valves, and high pressure carbon dioxide supply. Because many existing municipal wastewater treatment plants are well suited for co-location or integration of an algal cultivation system, the project team's knowledge about how treatment plants operate will be valuable in implementing other algal wastewater systems. Conducting a pilot plant in a real operating wastewater treatment plant can be considerably different than performing bench-top studies in a laboratory. How have the results been disseminated to communities of interest?The results of our work have not been disseminated to the public at large. We have connected with USDA's Forest Products Laboratory in Madison, Wisconsin to share our algal cellulose knowledge and with two academic institutions Purdue University and Oregon State University introduced to us by FPL for the same. The municipal wastewater treatment plants are a community of interest because they may be directly affected by implementation of our algal technology. For this reason, we have been disseminating information from our algal pilot plant to Madison Metropolitan Sewerage District. Project employee M. Piotrowski organized and delivered an activity on algal polysaccharides designed for young children during a "Darwin Day" public outreach event organized by the University of Wisconsin-Madison. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The growth experiments conducted proved that Oedogonium, a freshwater filamentous green alga (FFGA) could be grown in secondary effluent of a municipal wastewater treatment plant. It showed that this FFGA could survive biological contamination and was easy to grow and harvest - all attributes presently lacking in commercial cultivation of microalgae. Raw cellulose was easily extracted from dried biomass and processed into high value crystalline cellulose. The results indicate that cultivation of FFGA could be integrated into existing treatment plants of all sizes, but particularly in small plants that serve communities in rural areas. These treatment plants would provide access to land required for cultivation at low price, and because they often use pond treatment system, i.e., waste stabilization ponds, existing ponds can be used to grow FFGA. Supplemental supply of carbon dioxide could be supplied by anaerobically digesting the settled sewage sludge and directing the exhaust gas from combustion of biogas (e.g., for facility heating purpose) to the cultivation ponds. The simplicity of cultivating and harvesting FFGA make this an appropriate technology for application in rural areas where it would not only provide sustainable environmental benefits, but also economic value from sale of cellulose. The overall objective of this project was to develop a low cost method of growing Oedogonium, a common green alga that is particularly high in cellulose. The specific issues this project addressed were the effects of controllable variables; inside or outside growth environment, degree of air mixing, and addition of carbon dioxide. The intended target audience included end-users of cellulose microcrystals and nanocrystals such as paper, paperboard, specialty papers, and filter manufacturers. Wastewater industry may be most immediately impacted by our technology due to algae cultivation in wastewater which removes nutrients, nitrogen and phosphorus. The results of experiments are summarized in Table A. Table A: Summary Data Obtained From Experimental Runs Column A B C D E F G Experiment No. Start Date Experimental Conditions Location and Reactor No. Harvest. growth Dry biomass at 15% (dry g) % of dry biomass as cellulose Extracted cellulose (dry g) Production of dry cellulose ( g m-2 d-1) Production of CN at 20% ( g m-2 d-1) No. 1 Start 8/13 Low Air Mixing Low CO2 Inside 1 107.1 16.07 35% 5.5 0.43 0.09 Inside 2 0.00 0.0 Inside 3 64.5 9.68 32% 3.1 0.24 0.05 Outside 4 220.1 33.02 52% 17.1 1.32 0.26 Outside 5 85.8 12.87 46% 5.9 0.46 0.09 Outside 6 296.8 44.52 27% 12.0 0.93 0.19 Mean 154.9 19.4 38% 7.3 0.67 0.13 No. 2 Start 8/30 Low Air Mixing Low CO2 Inside 1 489.3 73.40 57% 41.6 4.81 0.96 Inside 2 414.6 62.19 58% 36.0 4.16 0.83 Inside 3 276.4 41.46 47% 19.6 2.27 0.45 Outside 4 617.6 92.64 34% 31.9 3.69 0.74 Outside 5 562.1 84.32 48% 40.5 4.69 0.94 Outside 6 502.3 75.35 50% 37.5 4.35 0.87 Mean 477.1 71.6 49% 34.5 3.99 0.80 No. 3 Start 9/16 High Air Mixing High CO2 Inside 1 603.6 90.54 31% 27.7 3.20 0.64 Inside 2 574.6 86.19 34% 29.0 3.36 0.67 Inside 3 643.0 96.45 32% 30.4 3.52 0.70 Outside 4 832.0 124.80 27% 33.4 3.87 0.77 Outside 5 880.4 132.06 32% 42.6 4.93 0.99 Outside 6 519.2 77.88 35% 27.6 3.20 0.64 Mean 675.5 101.3 32% 31.8 3.68 0.74 No. 4 Start 10/14 High Air Mixing High CO2 With Diffuser Inside 1 521.3 78.20 32% 25.0 1.93 0.39 Inside 2 390.0 58.50 33% 19.2 1.48 0.30 Inside 3 277.0 41.55 24% 10.1 0.78 0.16 Outside 4 463.7 69.56 35% 24.5 1.89 0.38 Outside 5 283.8 42.57 36% 15.4 1.19 0.24 Outside 6 215.9 32.39 36% 11.5 0.89 0.18 Mean 358.6 53.8 33% 17.6 1.36 0.27 No. 5 Start 11/04 High Air Mixing High CO2 With Diffuser Inside 1 163.0 24.45 38% 9.3 0.72 0.14 Inside 2 120.2 18.03 34% 6.0 0.47 0.09 Inside 3 74.0 11.10 32% 3.6 0.28 0.06 Outside 4 200.4 30.06 36% 10.7 0.83 0.17 Outside 5 61.4 9.21 33% 3.1 0.24 0.05 Outside 6 135.3 20.30 34% 6.9 0.53 0.11 Mean Inside and Outside 125.7 18.86 35% 6.6 0.51 0.10 Mean Inside 314.6 47.2 34% 17.7 1.84 0.37 Mean Outside 391.8 58.8 37% 21.4 2.20 0.44 Objective No. 1: It was determined that cultivation in greenhouse did not result inincreased biomass productivity or in higher quality biomass. Objective No. 2: As result of adding low levels of CO2 biomass yield increased from mean of 154.9 grams in Run No. 1 to 477.1 grams in Run No. 2. The benefits of adding CO2 depends ultimately on the price of cellulose end products. Objective No. 3: As a result of adding high levels of CO2, biomass yield increased from mean of 477.1 grams in Run No. 2 to 675.5 grams in Run No. 3. Observation is made that although biomass growth was higher in Run No. 3, average percentage of cellulose extracted of 49% is higher for Run No. 2. This results in Run No. 2 yielding slightly higher quantity of extracted cellulose. Objective No. 4: In Run No. 4, joint effects of high mixing and high CO2 was investigated. Addition of CO2 was also done using a fine stone diffuser. The expectation of highest biomass yield did not materialize primarily due to lower temperature. Biomass yield of 358.6 grams in Run No. 4 was lower than in Run No. 3, 675.5 grams. In general, high CO2 additions affects the percentage of cellulose by biomass dry weight (p-value = 0.0001), and interaction effect with the growth period (8 days vs. 12 days). It suggests that optimal biomass yield with the highest cellulose percentage is dependent on growth period. Due to interaction effect, CO2 plays an integral role in cellulose formation. Also of note, high level of CO2 aids in maintenance of high quality Oedogonium biomass as CO2 tends to minimize contaminants in cultures. More research is required to determine the exact nature of these interactions and the effects of the various drivers for biomass productivity. Decreasing temperatures from experiment 1 through experiment 5 affected biomass productivity that masked the effects of elevated CO2 and of growth period. Objective No. 5: Cultivation using higher nutrient loading was not performed because wastewater source at the pilot plant site was limited to final disinfected effluent. Objective No. 6: Cellulose was extracted from dried algal biomass using the procedures described in the workplan. During the extraction process several areas were identified where the process could be modified to reduce costs. The NaClO2 was recovered, and reused up to two times, before needing to be discarded. Bleaching is requires for processing into pure cellulose, without further consideration for nanoparticle processing, or characterization. The NaOH removes hemicellulose from the final cellulose pulp, and rapidly neutralizes the bleach. Thus, for the production of high purity cellulose from the Oedogonium, and other FFGA biomass, it may not be necessary undergo the NaOH treatment step, thus minimizing fine chemicals needed for the biomass processing. Objective No. 7: Inspection of processed Oedogonium cellulose using Raman microscopy indicated absence of crystals. As a result of this finding, conversion of Oedogonium cellulose into Crystalline Nanocellulose was not performed. In the future, we intend on heat treating Oedogonium cellulose as is done with wood cellulose. Heat treatment may yield nanocellulose crystals from Oedogonium. Explanation of qualitative and quantitative experimental data summarized in Table A areavailable, but could not be presented due to character limitation.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2015
Citation:
Graham, LE, JJ Knack, LW Wilcox. 2015. Why we need more algal metagenomes. Journal of Phycology 51:1029-1036.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2015
Citation:
Graham, LE, JJ Knack, ME Graham, JM Graham, S Zulkifly. 2015. A metagenome for lacustrine Cladophora (Cladophorales) reveals remarkable diversity of eukaryotic epibionts and genes related to materials cycling. Journal of Phycology 51:408-418.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2015
Citation:
Lee, PO, SL McClellan, LE Graham, EB Young. 2015. Invasive dreissenid mussels and benthic algae in Lake Michigan: Characterizing effects on sediment bacterial communities. FEMS Microbiology Ecology 91.
- Type:
Books
Status:
Accepted
Year Published:
2015
Citation:
Graham L., Graham J., Wilcox L., and Cook M, Algae 3rd Edition, LJLM Press
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