Progress 07/01/20 to 02/28/21

Outputs
Target Audience:The development of new cultivation systems for these new macroalgal species will offer new opportunities for sustainable and environmentally responsible economic development in the US, especially in rural coastal areas. Economically viable seaweed farming can offer opportunity for diversification to traditional fishermen, oyster, mussel, and fish farmers, and encourage more women farmers. Seaweed farming can help improve water quality, requires no fertilizer or fresh water, and produces nutritionally dense vegetarian seafood. The new crops can be sold fresh, dried, and processed into other snacks and foods, creating new opportunities for farmers, food producers, and consumers. Changes/Problems: Challenges to the work have included some degree of microalgal contamination in cultures utilizing only ultra-filtration strategies to clean seawater. An ultra-fine tangential flow filter became available to our facility after initiation of this project, so it was trialed during cultivation establishment as an alternative method for seawater sterilization. Despite the 0.01 micron filration capability, brown and green micro and macro algal contaminants were still present in cultures after several months. Some of the cultures will have to be re-initiated utilizing the electrochemically treated seawater. What opportunities for training and professional development has the project provided? The three interns involved in the project have worked closely with PI on all aspects of nursery cultivation, and are developing nursery and farm cultivation skills. How have the results been disseminated to communities of interest?Our seaweed facility has hosted several field trips from students from the College of the Atlantic and the University of Maine. Students from the College of the Atlantic were loaned cultivation supplies from our facilty, with guidance, for the establishment of an experimental seaweed culture nursery at their campus. Students have been encouraged to partner with our company when designing independent studies or internships. The PI has given several webinars on seaweed aquaculture to audiences at the University of Maine, the Maine Center for Coastal Fisheries, the University of Massachussetts, and to WL Gore and Associates. An international professional dialouge has been established between the PI and a nori researcher at CEVA, an algal research institution in France. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? There is increasing interest from aquaculture industries and markets for cultivated red seaweeds, especially dulse and nori, however, there is currently no commercial source of seed for cultivation, or cultivation technologies for farmers available in the U.S. or Europe. The purpose of this SBIR Phase I study was to develop critical nursery technologies and techniques for reliable production of dulse and nori seed, laying the groundwork for commercial nursery production of seed for our farm and other growers. Springtide Seaweed's goal in Phase I of this project was to develop reliable nursery protocols for commercial production of seed for dulse and nori, which will support the development of a new seaweed farming industry for the North Atlantic. Solutions to specific bottlenecks in nursery production were targeted, including the testing of an innovative approach to seawater sterilization utilizing electrochemical treatment technology, the development of nursery "seedstock" for the reliable production of spores, and the production of nori and dulse seed for open farm cultivation. Electrochemical Water Treatment: A new seawater treatment system was tested for use in seaweed production, utilizing electrochemical treatment of natural seawater to reduce contamination in the nursery. Electrochemical treatment of seawater is a process that passes seawater through a low voltage electro catalyst reactor, exposing organisms to electric field, electrooxidizing, and residual oxidant stress. Major findings from Phase I work found that electrochemical treatment of filtered seawater is an effective means to sterilize incoming seawater during exposure, effectively eliminating ciliates, diatoms, and other algal species at all levels tested (200, 500, 800, and 1500mA). Very small amounts of 35% food grade hydrogen peroxide is effective at neutralizing residuals in the water, as well as activated carbon water filters. Nori and Dulse Seedstock: New strains of dulse and nori were isolated and cultivated in the nursery as seedstock was developed for clean seed production. A major finding was that strain selection will be critically important for nursery domestication and development of higher yielding crops. Two types of dulse, intertidal and subtidal, were observed to have different morphologies and levels of biofouling, but were both stimulated to produce spores in stimulating environmental conditions in only 2 months. Dulse nursery seedstock cultures over 1 year old were also stimulated to release spores, and dulse and nori held under low light and freezing conditions recovered, indicating that there are a number of options to hold seedstock and seed in the nursery over long periods of time. Two nori strains were collected in the lower intertidal, a wide and a linear form, that adapted to tank culture much better than the higher intertidal strain. All strains released neutral spores over the course of the project that were readily seeded onto substrates. The conchocelis form of Wildemania amplissima was successfully established in a wide range of shells tested, as well as in advanced ePTFE substrate. Dulse Seeding Experiments: Dulse spores were seeded in the nursery in August ("early") and in September ("late") on a control (Kuralon, a Japanese kelp culture string) and advanced substrate test string utilizing wild collected tetrasporophytes of both intertidal and subtidal plants. The "early" seeded cages produced plants that were 3 times greater in length and 7 times greater in weight than the later seeded plants. Samples taken of both late and early set substrates in April indicate that advanced substrates support significantly better growth of dulse plants with less biofouling than the control. These initial farming results indicate potential to increase size and yield of farmed dulse through; 1) Early seeding in the nursery, for larger juvenile seed size at out-planting; 2) Strain selection utilizing larger plants as seedstock; and 3) Utilizing advanced substrates to reduce biofouling issues commonly experienced in nurseries and on open ocean farm sites. Phase I research enabled the development of innovative new approaches to seaweed nursery challenges and bottlenecks. By determining effective and safe electrochemical seawater treatment levels for seaweed culture, we have introduced a powerful new tool for natural seawater sterilization for algal culture. We discovered several different native nori and dulse strains that are amenable to cultivation, allowing us to focus on strain selection and domestication. While seedstock and nursery systems are critical for new crop development, cultivation design and suitable substrates are just as essential for successful seeding, grow-out, and harvest of new crops. Advanced substrates showed promising results for cultivation of dulse, resulting in larger plants, higher yields, and less biofouling compared to the control. Phase II will continue the work by refining species-specificsubstrates and cultivation systems for full farm grow out of the new nori and dulse species, utilizing strains, seedstock, and seawater sterilization methods developed in Phase I.

Publications

Progress 07/01/20 to 02/28/21

Outputs
Target Audience:Target audience reached for the reporting period include the general public, corporate partners, interns, college students, and recently graduated students. 3 interns have gained hands on experience with the work, 15 visitors (college students) have gained knowledge of the project and its goals through in-person field trips to our facility, and a number of students, fishermen, the general public, and corporate partners have gained knowledge through online meetings and presentations. Changes/Problems:Challenges to the work have included some degree of microalgal contamination in cultures utilizing only ultra-filtration strategies to clean seawater. An ultra-fine tangential flow filter became available to our facility after initiation of this project, so it was trialed during cultivation establishment as an alternative method for seawater sterilization. Despite the 0.01 micron filration capability, brown and green micro and macro algal contaminants were still present in cultures after several months. Some of the cultures will have to be re-initiated utilizing the electrochemically treated seawater. Additions to the project include the partnership with the advanced substrate company WL Gore and Associates. WL Gore has partnered with Springtide Seaweed to develop unique seaweed cultivation substrates for seaweed aquaculture. We are testing several of their unique substrates as part of our nori and dulse nursery and seed development research work, with the intention of developing specific culture systems for nori and dulse for the aquaculture industry. The emerging nature of this work requires unique designs specific to the needs of each crop, and our work with WL Gore is allowing us access to state-of-the-art substrates and technology. What opportunities for training and professional development has the project provided?The three interns involved in the project have worked closely with PI on all aspects of nursery cultivation, and are developing nursery and farm cultivation skills. How have the results been disseminated to communities of interest?Our seaweed facility has hosted several field trips from students from the College of the Atlantic and the University of Maine. Students from the College of the Atlantic were loaned cultivation supplies from our facilty, with guidance, for the establishment of an experimental seaweed culture nursery at their campus. Students have been encouraged to partner with our company when designing independent studies or internships. The PI has given several webinars on seaweed aquaculture to audiences at the University of Maine, the Maine Center for Coastal Fisheries, the University of Massachussetts, and to WL Gore and Associates. An international professional dialouge has been established between the PI and a nori researcher at CEVA, an algal research institution in France. What do you plan to do during the next reporting period to accomplish the goals?Experiments are ongoing for the project, and will continue through the end. Early nursery work will be repeated, due to microalgal contamination utilizing only ultra-fine sterilization without electrochemical treatment. Further experimentation on electrochemical treatment of seawater and effects on target organisms and crops will continue, as well as strain selection and seedstock establishment and stimulation. Seed will continue to be monitored for succes on the farm after out-planting. Further cultivation experiments are being conducted on unique advanced substrates with WL Gore and Associates for the development of species-specific seaweed cultivation substrates and growing systems for dulse and nori. Plans for commercialization are underway, as results from this project informs cultivation systems and strategies for nursery and farm production for our operation, as well as for licensing systems for global distribution.

Impacts
What was accomplished under these goals? The potential for sea vegetable aquaculture in North America and Europe is immense, and recognition of this potential has been growing in recent years.Currently seaweed aquaculture in NorthAmerica is limited to kelp production, which has alimitedmarketplace. The red seaweeds dulse (Palmaria palmata) and nori (Porphyra spp.) are highly nutritious, highly valued, rare native sea vegetables from the North Atlantic, making them species of interest for aquaculture. While there is increasing interest from both farmers and buyers for dulse and nori, there is no commercial source of seed or commercial farming cultivation systems for these species. We are developing reliable nursery and farming cultivation systems for production of nori and dulse seed and crops for North America and Europe. Seaweed aquaculture is a sustainable and environmentally responsible form of food production, requiring no fresh water, fertilizers, pesticides, herbicides, or arable land, and very little fossil fuel, and can be USDA organically certified. The development of new crops in aquaculture can contribute to our working waterfronts and marine heritage by providing new opportunities for diversification in the fishing communities, supporting rural coastal areas, and producing healthy sustainable seafood. Springtide Seaweed's goal is to develop reliable nursery protocols for commercial production of dulse and nori. Objective 1: Develop and test electrochemical water treatment strategies for the production of clean seawater for nursery cultivation. 1) Major activities completed / experiments conducted: Preliminary work with electrochemical water treatment has been conducted on neutralization strategies, and on ciliate contaminated solutions. Further work is being conducted on elimination of green and brown algal contaminants and on effects of treated seawater on cultivation of nori and dulse micro and macro stages. 2) Data collected: Data collected on ORP and free chlorine residuals before and after treatments, and photos taken for future analysis of effects of low, medium, and high treatments on cilitate solutions. 3) Summary statistics and discussion of results: Data is still being collected and analyzed for electrochemical treatment experiments. Preliminary results indicate that neutralization after treatment can be achieved naturally over time with circulation of the treated water, and more quickly with a carbon filter. All treatment levels resulted in neutralization of ciliates. 4) Key outcomes or other accomplishments realized. Electrochemical treatment can be an effective means to remove unwanted contamination in seawater systems. Several other filtration methods were trialed alongside the electrochemical treatment strategy, and all resulted in some form of brown or green algal contamination inside the water holding and cultivation systems, indicating the need for a more thorough treatment strategy offered by electrochemical sterilization. Objective 2: Establish clean nursery reared dulse and nori seedstock for the production of spores. 1) Major activities completed / experiments conducted: Development of new cultivation chambers and systems for dedicated R&D for nori and dulse cultivation and seed production; Collection and preliminary establishment of new strains dulse and nori for seedstock development and seed production; and vegetative and conchocelis seedstock establishment of nori and dulse. 2) Data collected: Data collected on strain collection, spore release, and establishment of new nursery strains. 3) Summary statistics and discussion of results: Strain development is still ongoing. Preliminary work indicates differences in sub tidally vs intertidally collected nori and dulse plants, and identification of favorable nori strains with direct seeding capabilities. 4) Key outcomes or other accomplishments realized: Strain selection for nursery culture will be an essential component to successfully establishing year round strains in cultivation. Dulse seedstock can be held indefinitely in culture conditions, though spore viability over time is not yet understood. There is a wide variety of nori species and strains in the wild, but most are not amenable to tank cultivation conditions over time. Strain selection for domestication will be a key element to successful seedstock establishment. Objective 3: To produce clean viable seed for open farm and tank cultivation. 1) Major activities completed / experiments conducted: Two groups of dulse seedstock (one year in cultivation, and one month in cultivation) are being cultivated at holding conditions, and two levels of spore-stimulating conditions (short day, and ultra-short day) to determine spore development of seedstock. Nori and Dulse seed were produced from field-collected plants this fall, and cultivated on several different advanced substrates to 1mm, when they were out planted on the open ocean farm for grow-out. 2) Data collected: Data collected on spore release and seed development, and monitoring is ongoing for farm growth and seedstock experiments. 3) Summary statistics and discussion of results: Seedstock initiation and holding experiments are ongoing. Nori and dulse seed were successfully produced on a variety of advanced cultivation substrates with promising anti-fouling results. 4) Key outcomes or other accomplishments realized: Research on nori and dulse seed production on advanced substrates with global corporate partner WL Gore and Associates have indicated that major obstacles in seed production can be overcome with advanced cultivation substrates. Research on new substrates for nori and dulse on these new Gore advanced materials is ongoing, with plans to develop cultivation systems specific to nori and dulse for the US and European markets.

Publications