2005 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? U.S. consumers demand a cost-competitive, safe, reliable animal protein supply protected from the vagaries of changing world events and environmental calamity. This food must also be appealing, nutritious, and raised with minimal environmental impacts. Controlled intensive aquaculture systems are intrinsically secure agriculture systems. Aquatic animals are produced in a semi-closed environment with a protected water supply. Inputs to the system can be controlled, so quality assurance is comparatively easier to achieve than in some other animal confinement systems. Controlled intensive aquaculture systems are poised to expand to a large role in the aquacultural production of the US domestic edible seafood supply. This project plan uses a multi-disciplinary approach to develop and evaluate solutions for major challenges that delay this expansion. The objectives of this plan are:. 1)To develop and evaluate solutions that improve efficiencies of scale and reduce water quality constraints for sustainable production in controlled intensive aquaculture systems. . 2) To develop and evaluate sustainable waste management technologies that result in environmentally compatible controlled intensive aquaculture systems. The capacity to produce a nutritious seafood product in an aquaculture system that is secure, reliable and economically and environmentally sustainable is the outcome expected from this work. Improvement in resource and capital efficiencies for controlled intensive aquaculture systems will result in superior production systems, improved management practices and expanded market and investment opportunities for domestic aquaculture production. This research program will result in more sustainable and globally competitive aquaculture systems for U.S. farmers. This work is relevant to consumers demanding cost competitive, high quality fish that have been raised in environmentally friendly production systems, fish farm operations producing a variety of freshwater and marine species in tank-based systems, and scientists and consultants that design and evaluate sustainable land-based finfish production systems. 2.List the milestones (indicators of progress) from your Project Plan. Objective 1. To develop and evaluate solutions that improve efficiencies of scale and reduce water quality constraints for sustainable production in controlled intensive aquaculture systems. Objective 2. To develop and evaluate sustainable waste management technologies that result in environmentally compatible controlled intensive aquaculture systems. FY 2005 Objective 1. Solutions for efficiencies of scale and water quality constraints 1.1. Overcome obstacles associated with large-scale culture tanks, the effect of noise in controlled intensive aquaculture systems • Install 600 m3 culture tank system, begin solids flushing studies, begin velocity profile studies, begin studies testing CO2 avoidance behavior for promoting fish transfer, begin evaluating culture tank noise 1.2. Improve production system water quality • Begin advanced oxidation studies, conduct descriptive study of rainbow trout health in high feed/low flow conditions, set up and evaluate CO2 stripping in marine controlled intensive aquaculture systems, collect growth and survival data on trout for NCCCWA 1.3. Mitigate pathogens originating in the water supply • Finish molecular characterization of CLO Objective 2. Sustainable waste management technologies and practices 2.1. Treat solids and nutrients in aquaculture effluents • Begin research on solids capture and dewatering in geotextile tubes 2.2. Mitigate impact of feed on effluent quality • Conduct trials using commercially available grain-based diets FY 2006 Objective 1. Solutions for efficiencies of scale and water quality constraints 1.1. Overcome obstacles associated with large-scale culture tanks, the effect of noise in controlled intensive aquaculture systems • Complete solids flushing studies, complete velocity profile studies, complete studies of CO2 avoidance for promoting fish transfer, begin studies testing low O2 avoidance behavior for promoting fish transfer, complete studies on culture tank noise. 1.2. Improve production system water quality • Continue advanced oxidation studies, build replicated recirculating systems, begin studying hypotheses re RBT health in high feed/low flow conditions, complete empirical studies of CO2 stripping in seawater systems, collect growth and survival data on trout for NCCCWA 1.3. Mitigate pathogens originating in the water supply • Develop CLO assay, investigate CLO infection reservoirs, Objective 2. Sustainable waste management technologies and practices 2.1. Treat solids and nutrients in aquaculture effluents • Complete research on solids capture and dewatering in geotubes, begin research on geotextile tubes as membrane biological treatment systems 2.2. Mitigate impact of feed on effluent quality • Conduct trials using comercially available grain-based diets FY 2007 Objective 1. Solutions for efficiencies of scale and water quality constraints 1.1. Overcome obstacles associated with large-scale culture tanks, the effect of noise in controlled intensive aquaculture systems • Compete studies testing low O2 avoidance behavior for promoting fish transfer, transfer technology culture tank noise. 1.2. Improve production system water quality • Continue advanced oxidation studies, continue studies of RBT health in high feed/low flow conditions, complete modeling of CO2 stripping in marine controlled intensive aquaculture systems, install O2 absorber/CO2 scrubber in existing partial reuse system, collect growth and survival data on trout for NCCCWA 1.3. Mitigate pathogens originating in the water supply • Characterize bacteria in fish GI tract Objective 2. Sustainable waste management technologies and practices 2.1. Treat solids and nutrients in aquaculture effluents • Continue research on geo-tubes as membrane biological treatment systems, begin research on MBR systems for nutrient removal 2.2. Mitigate impact of feed on effluent quality • Conduct trials using Barrow’s grain-based diets FY 2008 Objective 1. Solutions for efficiencies of scale and water quality constraints 1.1. Overcome obstacles associated with large-scale culture tanks. • Evaluate clamshell crowder/grader on 600 m3 culture tank 1.2. Improve production system water quality • Continue advanced oxidation studies, continue studies of RBT health in high feed/low flow conditions, evaluate O2 absorber/CO2 scrubber in existing partial reuse system, collect growth and survival data for trout for NCCCWA 1.3. Mitigate pathogens originating in the water supply • Finish characterizing bacteria in fish GI tract, begin testing bacteriophage, Objective 2. Sustainable waste management technologies and practices 2.1. Treat solids and nutrients in aquaculture effluents • Complete research on geo-tubes as membrane biological treatment systems, continue research on MBR systems for nutrient removal 2.2. Mitigate impact of feed on effluent quality • Conduct trials using Barrow’s grain-based diets FY 2009 Objective 1. Solutions for efficiencies of scale and water quality constraints 1.1. Overcome obstacles associated with large-scale culture tanks • Complete studies on fish transfer systems 1.2. Improve production system water quality • Continue advanced oxidation studies, finish studies of RBT health in high feed/low flow conditions, evaluate O2 absorber/CO2 scrubber in existing partial-reuse system, collect growth and survival data on trout for NCCCWA 1.3. Mitigate pathogens originating in the water supply • Finish testing phage and transfer technology Objective 2. Sustainable waste management technologies and practices 2.1. Treat solids and nutrients in aquaculture effluents • Complete research on MBR systems for nutrient removal 2.2. Mitigate impact of feed on effluent quality • Finish trials using Barrow’s grain-based diets FY 2010 Objective 1. Solutions for efficiencies of scale and water quality constraints 1.1. Overcome obstacles associated with large-scale culture tanks. • Transfer technology 1.2. Improve production system water quality • Complete advanced oxidation studies, transfer technology for results rainbow trout health in high feed/low flow conditions, complete studies on O2 absorber/CO2 scrubber in existing partial-reuse system, collect growth and survival data on trout for NCCWA Objective 2. Sustainable waste management technologies and practices 2.1. Treat solids and nutrients in aquaculture effluents • Transfer technology 2.2. Mitigate impact of feed on effluent quality • Transfer technology 4a.What was the single most significant accomplishment this past year? New project, cooperative agreement initiated 6/15/2005 4b.List other significant accomplishments, if any. Noise reduction in fish tanks Noise is generated in aquaculture systems that utilize mechanical components. Fish exposed to extreme levels of noise may be stressed, grow more slowly and be more susceptible to disease. The noise reduction potential of various retrofits to fiberglass fish culture tanks were measured. The optimal retrofit combination that most effectively minimized in-water sound pressure levels (SPL’s) included suspended inlet spray bars, sound insulation under the tank bottom and disconnection from the common drain line. This research was the result of a collaboration between The Conservation Fund’s Freshwater Institute (Shepherdstown, WV), Marine Acoustics, Inc. (Annapolis, MD) and the University of Maryland Laboratory of Aquatic Bioacoustics (College Park, MD). Reduction of in-water noise in the culture tank will allow engineers to improve fish well-being and productivity through system design changes. 4c.List any significant activities that support special target populations. Freshwater Institute scientists have provided technical support in aquaculture engineering, fish health and biosecurity, and trout and Arctic char culture across the Appalachian region. This work supports economic development in a region where many counties are rated as economically distressed and unemployment rates are greater than 10%. Across the Appalachian states of West Virginia, Kentucky, Virginia, Maryland and Pennsylvania, there are many natural resources that appear to represent potential development opportunities. Current production figures indicate that the Mid-Atlantic Highlands region has not yet participated in the general expansion of the aquaculture industry in the U.S. 4d.Progress report. This report documents research conducted under an Assistance Type Cooperative agreement between ARS and the Conservation Fund’s Freshwater Institute. Additional details of research can be found in the report for the parent CRIS 1930-32000-003-00D, Development of Sustainable Land-based Aquaculture Production Systems and prior CRIS 1930-31000-004-00D. Voluntary fish transfer systems We have begun testing whether a fish’s ability to sense and seek to avoid areas containing higher concentrations of dissolved carbon dioxide or relatively low concentrations of dissolved oxygen can be used to induce the fish to voluntarily swim out of a culture tank during a harvest event. The study system was installed using two 3.7 m diameter (11.1 m3 volume at 1.07 m depth) culture tanks that are located in a single-pass system at the Conservation Fund Freshwater Institute. The adjacent tanks were connected with a 203.2 mm diameter fish transfer channel. Both tanks are supplied with oxygenated water and their own side-stream down-flow contactor for transferring oxygen and carbon dioxide gas. A system that allows for adjusting dissolved carbon dioxide and dissolved oxygen concentrations within one of the culture tanks has been installed. Controlled and replicated experiments were begun to determine if use of 60, 120, or 240 mg/L dissolved carbon dioxide (at a constant dissolved oxygen concentration of approximately 100% saturation) could induce fish to swim from this environment into a channel or pipe that is flowing water containing a dissolved carbon dioxide concentration of ¿ 10 mg/L and a dissolved oxygen concentration of at least 100% saturation. The fraction of the total number of fish that swim out of the high dissolved carbon dioxide (or low dissolved oxygen) tank within a 2 hr period is being recorded for each condition. Each condition is being replicated three times. Carbon dioxide stripping in seawater Anecdotal evidence from commercial marine fish farmers indicates that dissolved carbon dioxide stripping across forced-ventilated cascade columns is more difficult in seawater than in freshwater and there is no published research describing carbon dioxide stripping column design and performance in seawater applications. Empirical studies were begun to determine how air:water loading levels and packing type affect dissolved carbon dioxide removal at different salinity levels in clean water. Dissolved carbon dioxide stripping efficiency was evaluated across two types of 1-m tall packing that were placed separately in two full-scale (0.91 m long x 1.22 m wide) forced-ventilation cascade columns located within a recirculating aquaculture system. Volumetric air:water loading rates that varied from 1:1 to 10:1 were produced when the water flow rates through each column were set at 1.19, 2.38, and 3.57 m3/min (315, 630, and 945 gpm), respectively. The resulting air flow rates through the columns were measured under these conditions. Salinity levels in the recirculating system were < 0.1 ppt during freshwater control trials, but are being set at 32 ppt during the high salinity trials. The dissolved carbon dioxide concentration supplied to the forced-ventilated cascade aeration columns was set at approximately 20 mg/L by diffusing pure carbon dioxide gas into the pump sump of the system. Studies are also being conducted at water temperatures of approximately 15¿C and 25¿C, to determine if water temperature affects carbon dioxide stripping. Bacteria inactivation with ultraviolet irradiation and ozone Research was substantially completed to determine the combined dosages of ozone and ultraviolet irradiation required to inactivate total heterotrophic and total coliform bacterial populations in a commercial-scale recirculating system used to produce food-size rainbow trout. One experimental set-up tested bacteria inactivation within a side-stream of water pumped from a sump in the recirculating system and the other set-up tested for bacteria inactivation in full-flow (4,800 L/min) ozonation within the system low head oxygenation unit followed by full-flow ultraviolet irradiation. The side-stream water flow was adjusted to either 150 or 300 L/min (i.e., approximately 3.2 and 6.4% of the entire recirculating flow) and then the concentration of ozone gas transferred was adjusted so as to produce three dissolved ozone treatment concentrations at the end of the contact basin, i.e., approximately 0.05, 0.10, and 0.20 mg/L. The sidestream of ozonated water then flowed through a 3,200 L u-tube contact column or was by-passed directly to a ultraviolet irradiation unit. The effect of noise on rainbow trout hearing, growth, survival and disease susceptibility. Sound frequencies and sound pressure levels within a commercial scale recirculating system were characterized. An audio recording that simulates these sound characteristics was created. A CD player, amplifiers, and tactile speakers were installed to transmit the audio recording through experimental tanks to provide three replicated sound treatment categories: 115, 130, and 150 dB re 1 ¿Pa (RMS). Rainbow trout were stocked in these tanks at 17 grams and will be cultured under these conditions until they reach average weights of 200 to 250 grams and maximum densities of 75 kg/m3. The auditory brainstem response (ABR) will be used to determine whether hearing loss occurs as a result of these sound levels. Growth is being measured every two weeks and the growth rates of fish from each sound treatment category will be compared. Feed conversion ratios, condition factors, and survival between sound treatments will also be compared. Response to a Yersinia ruckeri pathogen challenge will be recorded. This work is a collaboration between The Conservation Fund’s Freshwater Institute (Shepherdstown, WV), the University of Maryland Aquatic Bioacoustics Laboratory (College Park, MD), Marine Acoustics, Inc. (Annapolis, MD), and the USDA-ARS National Center for Cool and Coldwater Aquaculture (Leetown, WV). Impaired rainbow trout health in systems that feed at greater than 1.3 kg/d feed per m3/d makeup water flow. A commercial scale recirculating system was run at >1.3 kg/d feed per m3/d makeup water flow and at <1.3 kg/d feed per m3/d makeup water flow, with and without ozone. Blood samples were collected to measure plasma chemistries, and plasma ammonia and cortisol. Water samples were collected to measure organics and other water quality characteristics. Data analysis is in progress. This work is a collaboration between The Conservation Fund’s Freshwater Institute (Shepherdstown, WV) and the USDA-ARS National Center for Cool and Coldwater Aquaculture (Leetown, WV). Molecular characterization of a chlamydia-like bacterium The following objectives regarding characterization of the chlamydia-like agent associated with epitheliocystis in Arctic char have been completed: (1) amplification of both the 16S SSU and the 23S LSU ribosomal DNA genetic sequences, and (2) in situ hybridization of the 16S signature sequence RNA probe. Phylogenetic analysis places this agent with members of the chlamydiales. This work is the result of a collaboration between The Conservation Fund’s Freshwater Institute and the University of Connecticut. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. New project 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? New project 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Bebak-Williams, J. 2005. Fish health management and biosecurity for recirculation aquaculture systems. Eleventh Annual Freshwater Institute-Cornell University Recirculating Aquaculture Systems Short Course. July 18-22, 2005, Hubbs-Seaworld Research Center, San Diego, CA. Bebak-Williams, J., Welch, T., Starliper, C., Baya, A., Garner, M. 2005. Coldwater disease outbreak at a West Virginia rainbow trout (Oncorhynchus mykiss) farm: An unusual presentation. 30th Annual Eastern Fish Health Workshop, June 13-17, 2005, Shepherdstown, WV, p. 64. Draghi, A., Bebak-Williams, J., Tsongalis, G.J., West, A.B, Popov, V.L., Frasca, Jr, S. 2005. Localization by in situ hybridization of a chlamydia-like 16S ribosomal RNA gene sequence within branchial epithelial cells of cultured Arctic char (Salvelinus alpinus) with epitheliocystis. 30th Annual Eastern Fish Health Workshop, Shepherdstown, WV, June 13-17, p. 85. Ebeling, J.M. 2005. Aquaculture Effluent Waste Management And Treatment. In: Stormwater and Aquaculture Effluent Technology Fair. March 8, 2005, NOAA Beaufort Lab and Duke University Marine Lab, Beaufort, NC. Lehman, M.C. 2005. The Freshwater Institute: Promoting Sustainable Economic Development Through Aquaculture. May, 2005, The Shepherdstown Observer, p. 18-19. Summerfelt, S.T. 2005. Culture tank design; Culture tank design examples; Biofiltration for ammonia control; Fluidized-sand biofilters; Biofilter design examples; Carrying capacity examples; Ozonation and UV disinfection; BMP’s for recirculating systems; Emergency response. Presented at the 11th Annual Cornell-Freshwater Institute Recirculating Aquaculture Systems Short Course, July 18-22, Hubb-Seaworld Research Center, San Diego, CA. Summerfelt, S.T. 2005.Water recirculating systems for salmonid production. Fisheries Committee Meeting, June 21, 2005, Pennsylvania Fish & Boat Commission, Harrisburg, PA. Timmons, M.B., Holder, J.L., Ebeling, J.M. 2005. Microbead Filters: cost-effective, scalable filtration. Global Aquaculture Advocate 8(1):68-69. Vinci, B.J. 2005. Engineering design concepts; Solids capture; Gas transfer and dissolved gas conditioning; CO2 stripping design example; Source water development; Waste management. The 11th Annual Cornell-Freshwater Institute Recirculating Aquaculture Systems Short Course, July 18-22, Hubbs-Seaworld Research Center, San Diego, CA.