2006 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? Why does it matter? Aquaculture is poised to become a major growth industry of the 21st century. With increasing seafood demand and declining capture fisheries, U.S. marine aquaculture production must increase. However, development and expansion of the U.S. mariculture industry is challenged by the high cost and limited availability of coastal land and water resources, effluent concerns, high production costs, restricted growing seasons, lack of sufficient quality seedstock, and a general lack of knowledge regarding reproduction, larviculture, and husbandry. Innovative, cost-effective technologies to produce marine fish in land-based systems must be developed to overcome these constraints. The goal of this research project is to design and test cost-effective, energy efficient aquaculture systems and management strategies capable of sustainable, year-round production of marine warm-water species in low salinity environments. The development of these systems and production strategies will reduce the U.S. trade deficit, increase job market opportunities for inland rural sectors and displaced farmers, increase crop diversity, and enhance food biosecurity. In collaborative studies with Harbor Branch Oceanographic Institution (HBOI), research will be conducted to develop and evaluate recirculating nursery and growout production systems for marine finfish. The designs will incorporate computer systems to monitor water and air temperatures, water quality, water flow, and energy use. Findings will allow aquaculture production of saltwater species in new and underutilized locations in the US, resulting in better use of land resources, and reducing the demand for imported aquaculture products. Specific objectives include:. 1)development of culture technology to produce marine or brackish water species in low salinity or freshwater;. 2)improve the energy efficiency and reduce production costs for enclosed aquaculture production systems;. 3)evaluate recirculating aquaculture technologies to improve production efficiency of intensive systems;. 4)develop reliable larviculture techniques for sustained production of marine fish seedstock; and. 5)develop improved diets and feeding strategies. Striped bass and their hybrids, black sea bass, and Florida pompano are the initial test species. Production level recirculating systems will be used to conduct nursery and growout trials in fresh and low-salinity environments. Stocking densities, feeding strategies, and water reuse processes will be evaluated for effects on growth rate, feed conversion, time to market, and overall production cost. Findings will be correlated with overall system efficiency to fully optimize proposed production systems. This research addresses the following target areas of National Program 106-Aquaculture:. 1)reproduction and early development, as it relates to year-round production of gametes, spawning induction, diet development for enhanced gamete quality, and early life stage development;. 2)growth, development, and nutrition, as it relates to determining nutrient availability, nutrient requirements, and developing alternative ingredients to fish meal;. 3)aquaculture production systems, as it relates to biosecurity, development of new biological and engineering designs, and reuse technologies; and. 4)sustainability and environmental compatibility, as it relates to water use and reuse, and technologies for greater reclamation. Establishment of new marine aquaculture technologies for inland locations will significantly impact the U.S. agricultural sector. Increased numbers of marine fish farming enterprises will provide jobs and support collateral industries including the agriculture sector, feed manufacturers, processors, and aquaculture supply companies. Wholesalers, retailers, and consumers will benefit from increased supplies of safe, quality seafood products. Expansion of the U.S. aquaculture industry will reduce the U.S. trade deficit, increase job market opportunities for inland rural sectors, increase crop diversity, and enhance food biosecurity. 2.List by year the currently approved milestones (indicators of research progress) Year 1 (FY 2005) Conduct economic and production system evaluations for low-salinity production of hybrid striped bass in recirculating systems. Year 2 (FY 2006) Develop reference research diets for Florida pompano to determine essential nutrient requirements. Develop techniques and procedures for optimizing Florida pompano spawning. Characterize early development stages of Florida pompano and black sea bass. Establish performance characteristics of recirculating aquaculture system components during low-salinity fish production. Establish shortened spawning cycles for Florida pompano and black sea bass. Determine optimal temperature, salinity, light and density to enhance egg and larval survival. Year 3 (FY 2007) Characterize black sea bass waste production and excretion. Establish fecal settling velocity criteria for Florida pompano and black sea bass. Establish lysine requirements for Florida pompano and black sea bass. Evaluate essential fatty acids necessary to optimize spawning success for Florida pompano. Establish total sulfur amino acid requirements for Florida pompano and black sea bass. Develop reference research diets for black sea bass to determine essential nutrient requirements. Establish nutrient availabilities from traditional and novel feed ingredients for Florida pompano and black sea bass. Evaluate essential fatty acids necessary to optimize spawning success for black sea bass. Establish optimal female to male ratios for the spawning of Florida pompano and black sea bass. Develop optimal nutrient enhancement of live feeds for larval marine fish. Characterize Florida pompano waste production and excretion. Establish performance characteristics of recirculating aquaculture system components during low-salinity fish production. Year 4 (FY 2008) Establish arginine requirements for black sea bass and Florida pompano at different salinities. Establish nutrient availabilities from traditional and novel feed ingredients for Florida pompano and black sea bass. Determine optimal essential fatty acid ratios for Florida pompano broodstock to optimize spawning success. Evaluate optimal broodstock diets for Florida pompano and black sea bass reproductive success. Establish recommendations for the use of beneficial bacteria for the enhancement of egg and larval survival. Establish performance characteristics of recirculating aquaculture system components during low-salinity fish production. Develop a preliminary model for recirculating aquaculture system design and production. Year 5 (FY 2009) Establish histidine and threonine requirements for Florida pompano and black sea bass. Establish nutrient availabilities from traditional and novel feed ingredients for Florida pompano and black sea bass. Determine optimal essential fatty acid ratios for black sea bass broodstock to optimize spawning success. Develop optimal feeding strategies for Florida pompano and black sea bass larval survival. Develop year-round spawning techniques for Florida pompano and black sea bass. Develop a refined model for recirculating aquaculture system design and production. 4a.List the single most significant research accomplishment during FY 2006. POMPANO AND BLACK SEA BASS LARVAL DEVELOPMENT: Early life stages of Florida pompano and black sea bass were characterized via determination of growth and development by scientists of the ARS Sustainable Marine Aquaculture Project in Ft. Pierce, FL. Although limited knowledge exists regarding the timeframe of various physical and developmental changes that marine finfish larvae undergo, characterization of early life stages provides information that can be utilized to develop management protocols such as day of first feeding, appropriate size of live feed organisms to be offered, and time of complete metamorphosis. Larval Florida pompano and black sea bass were collected daily from culture tanks (from one day after hatching through complete metamorphosis) and microscopic images were captured and analyzed to determine parameters including length, yolk absorption rate, time of complete yolk absorption and first feeding, mouth size and development, and time of complete metamorphosis. This accomplishment will be used by fish hatchery managers to establish developmentally appropriate management methods for larval rearing of both species. This accomplishment addresses Year 2 (2006) milestone, characterize early development stages of Florida pompano and black sea bass, and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. 4b.List other significant research accomplishment(s), if any. DIETARY PROTEIN AND ENERGY FOR POMPANO: Optimal dietary protein and energy values for rearing juvenile Florida pompano were determined by scientists of the ARS Sustainable Marine Aquaculture Project in Ft. Pierce, FL. Fish feed represents the highest operating cost at aquaculture facilities with protein accounting for the greatest percentage of this cost. Identifying optimal protein and energy levels for Florida pompano will decrease feed costs, reduce time to market, and increase feed efficiency while optimizing product quality, increasing food safety, and maximizing profits for fish culturists. Florida pompano were fed diets containing a wide range of protein and energy levels, with fish receiving moderate levels of protein and high levels of energy performing as well as or better than fish receiving higher levels of protein and less energy. This accomplishment will be used by fish feed manufacturers to make more efficient diets with lower protein levels creating a significant savings to feed manufacturers and fish culturists alike. This accomplishment addresses Year 2 (2006) milestone, develop reference research diets for Florida pompano, and the following component of NP 106-Aquaculture: Growth, Development, and Nutrition. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, Performance Measures 1.2.1, 1.2.2, and 1.2.5. NUTRIENT DIGESTIBILITY IN POMPANO: Protein, energy, and other nutrient digestibility (availability) values were determined for six protein sources that can serve as alternatives to fish meal for use in Florida pompano diets. High quality fish meal is the best source of protein for fish; however, replacement of fish meal with alternative protein sources will increase sustainability and profitability of the aquaculture industry. Three plant-based (soybean meal, soy protein concentrate, and corn gluten meal) and three waste by-product-based (poultry by-product meal, meat and bone meal, and distillers dried grains) feed ingredients were fed to Florida pompano, and availability of protein, energy, and other nutrients were determined by scientists of the ARS Sustainable Marine Aquaculture Project in Ft. Pierce, FL. This accomplishment will be used by fish feed manufacturers to substitute these novel and traditional feed ingredients for fish meal resulting in efficient, less expensive, and more environmentally responsible diets. This accomplishment addresses Year 2 (2006) milestone, develop reference research diets for Florida pompano, and the following component of NP 106-Aquaculture: Growth, Development, and Nutrition. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, Performance Measures 1.2.1, 1.2.2, and 1.2.5. BLACK SEA BASS BREEDING: A breeding and larviculture program for mass production of black sea bass juveniles was established by scientists of the ARS Sustainable Marine Aquaculture Project in Ft. Pierce, FL. A major factor limiting marine finfish aquaculture is the lack of techniques for sustained and reliable production of seedstock to supply grow-out operations. Captured black sea bass were induced to spawn, eggs were incubated and hatched, and the resulting larvae were reared through metamorphosis to produce juveniles. Results identified potential bottlenecks regarding broodstock acquisition and holding, while establishing baseline procedures for the successful mass production of juvenile black sea bass. Outcomes of this accomplishment will enhance U.S. competitiveness in marine fish culture through development of technologies for year-round seedstock production, enhanced biosecurity, and domesticated strains of broodstock to produce quality black sea bass seedstock. This accomplishment addresses Year 2 (2006) milestone, develop techniques and procedures for optimizing black sea bass spawning, and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. POMPANO AMMONIA AND NITRITE TOLERANCE: The tolerance of early life stages of Florida pompano to ammonia and nitrite was determined by scientists of the ARS Sustainable Marine Aquaculture Project in Ft. Pierce, FL. Ammonia and nitrite constitute the two major waste products encountered in recirculating aquaculture systems, and knowledge of the levels detrimental to various life stages of fish is necessary for efficient management of culture systems. Eggs and larval Florida pompano of different ages (1, 8, and 16 days post-hatch) were subjected to short-term exposure (24 h) to varying concentrations of either ammonia or nitrite, and the tolerance of each life stage was determined. This accomplishment establishes the relative sensitivity of early developmental stages of pompano to ammonia and nitrite, and this information can be used by hatchery managers to avoid detrimental concentrations of these aquaculture waste products. This accomplishment addresses Year 2 (2006) milestone, determine optimal temperature, salinity, light and density to enhance egg and larval survival, and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. POMPANO SENSITIVITY TO COPPER: The sensitivity of early developmental stages of Florida pompano exposed to copper was determined by scientists of the ARS Sustainable Marine Aquaculture Project in Ft. Pierce, FL. While copper is an effective treatment against various pathogens that can cause mass mortality of aquacultured fish, limited information exists with respect to the tolerance of early life stages of marine finfish. Eggs and larval Florida pompano of different ages (1, 8, and 16 days post-hatch), as well as post-metamorphic juveniles, were subjected to short-term exposure (24 h) to varying concentrations of copper and the tolerance of each life stage was determined. This accomplishment establishes the relative sensitivity of early developmental stages of pompano to copper allowing hatchery managers to effectively implement appropriate disease treatment protocols. This accomplishment addresses Year 2 (2006) milestone, determine optimal temperature, salinity, light and density to enhance egg and larval survival, and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. SALINITY EFFECTS ON LARVAL POMPANO: The effect of environmental salinity on early life stages of Florida pompano was determined by scientists of the ARS Sustainable Marine Aquaculture Project in Ft. Pierce, FL. Non-scientific observations suggest Florida pompano are tolerant to a wide range of environmental salinities; however, limited information exists with respect to salinity effects on early developmental stages of pompano. Eggs and larval Florida pompano of different ages (1, 8, and 16 days post-hatch), as well as post-metamorphic juveniles, were subjected to short-term exposure (24 h) to varying salinity levels and the effects of salinity on each life stage was determined. This accomplishment establishes the range of salinities supporting acceptable egg buoyancy and hatching, as well as larval and juvenile survival and will increase the success rate and production efficiency of hatchery operators. This accomplishment addresses Year 2 (2006) milestone, determine optimal temperature, salinity, light and density to enhance egg and larval survival, and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. MEASURING DISSOLVED CARBON DIOXIDE: Three methods for measuring dissolved carbon dioxide (CO2) in a marine recirculating aquaculture system were developed by scientists of the ARS Sustainable Marine Aquaculture Project in Ft. Pierce, FL.. There are compounds and salts that can interfere with the determination of dissolved carbon dioxide concentrations in marine aquaculture systems; however, if the dissolved carbon dioxide gas can be released from the water it can be measured directly with a gas meter. Water at three salinities had three concentrations of dissolved carbon dioxide gas infused into them and the dissolved carbon dioxide gas in each combination of solutions was measured by three methods. This accomplishment indicates that removing carbon dioxide gas from water and measuring it directly with a portable hand-held gas analyzer provides a reliable and cost-effective method for fish culturists to measure CO2 concentrations in saltwater aquaculture environments. This accomplishment addresses Year 2 (2006) milestone, establish performance characteristics of recirculating aquaculture system components during low-salinity fish production, and the following components of NP 106-Aquaculture: Aquaculture Production Systems; and Sustainability and Environmental Compatibility. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. BIOFILTER NITRIFCATION: Rate of ammonia removal and performance curves for a fixed-film biofilter with floating bead media were determined by scientists of the ARS Sustainable Marine Aquaculture Project in Ft. Pierce, FL. There is no standard method for determining and reporting biofilter performance for the purpose of comparing biofiltration efficiencies; therefore determining ammonia removal rates would provide criteria to facilitate selection among the different types of biofilters. Four recirculating aquaculture systems with two fixed-film biofilters, each containing a different type of floating plastic bead media, received one of two different ammonia concentrations to evaluate ammonia removal at salinities ranging from 3 g/L to 15 g/L. This accomplishment provides aquaculturists, design engineers, and filter manufacturers the criteria needed to make well informed decisions for selecting biofilters for use over a wide range of salinities. This accomplishment addresses Year 2 (2006) milestone, establish performance characteristics of recirculating aquaculture system components during low-salinity fish production, and the following components of NP 106-Aquaculture: Aquaculture Production Systems; and Sustainability and Environmental Compatibility. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. AQUACULTURE INDUSTRY SURVEY: A survey of aquaculture industry experiences with different types of biological filtration was conducted by scientists of the ARS Sustainable Marine Aquaculture Project in Ft. Pierce, FL. Recirculating technology for aquaculture relies almost exclusively on biofilters for ammonia removal and although many types of biofilters are commercially available, requirements for use in recirculating systems have not been systematically quantified. A survey was conducted canvassing over 400 people in the aquaculture industry including scientists and technicians at universities; private, public, and government research facilities; State and Federal hatchery managers; and aquaculture farmers for the purpose of quantifying filtration needs. This accomplishment provides biofilter manufacturers the demographics, requirements, and needs specific applications of biofilter users in recirculating aquaculture. This accomplishment addresses Year 2 (2006) milestone, establish performance characteristics of recirculating aquaculture system components during low-salinity fish production, and the following components of NP 106-Aquaculture: Aquaculture Production Systems; and Sustainability and Environmental Compatibility. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. 4c.List significant activities that support special target populations. None. 5.Describe the major accomplishments to date and their predicted or actual impact. This project is a cooperative research program between ARS and Harbor Branch Oceanographic Institution (HBOI), Fort Pierce, FL, and is in its second year. Major accomplishments of the project include: POULTRY BY-PRODUCT DIETS: Hybrid striped bass were successfully produced in production-scale recirculating systems with diets containing poultry by-product meal as a fish meal replacement. Although high quality fish meal is the best source of protein for fish, replacement of fish meal with suitable alternative protein sources will increase environmental sustainability and profitability of the U.S. aquaculture industry. The Stuttgart National Aquaculture Center in collaboration with Harbor Branch Oceanographic Institution demonstrated hybrid striped bass can be reared throughout a growout cycle in recirculating systems with feeds containing 35% replacement of fish meal with poultry by-product meal while obtaining comparable growth and efficiency relative to a standard hybrid striped bass commercial diet. The results of this investigation will allow aquaculture feed manufacturers to produce hybrid striped bass diets that will convert waste material from the poultry industry into a salable high-quality and nutritious protein source at lower cost and in an environmentally sustainable manner. This accomplishment addresses Year 2 (2006) milestone, establish performance characteristics of recirculating aquaculture system components during low-salinity fish production and the following components of NP 106-Aquaculture: Growth, Development, and Nutrition; and Aquaculture Production Systems. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, Performance Measures 1.2.1, 1.2.2, and 1.2.5. POMPANO BROODSTOCK PROGRAM: A Florida pompano broodstock domestication program was initiated by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL. Lack of techniques for sustained production of seedstock for grow-out operations is one of the foremost bottlenecks of marine finfish aquaculture, and broodstock domestication is known to enhance seedstock quality. New broodstock were subjected to quarantine procedures, feed training, and conditioning for spawning. The outcomes of this program will enhance U.S. competitiveness in marine fish culture through development of technologies for year-round seedstock production, enhanced biosecurity, and domesticated strains of broodstock to produce quality Florida pompano and black sea bass seedstock. This accomplishment addresses Year 1 (2005) milestone, develop techniques and procedures for optimizing Florida pompano spawning and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. EVALUATION OF RECIRCULATING AQUACULTURE SYSTEM COMPONENTS: Efficiencies of solid waste and ammonia removal were determined by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL for various recirculating aquaculture system components in production scale recirculating systems used to rear hybrid striped bass and tilapia to market size. Expansion of the U.S. marine aquaculture industry is challenged by the high cost and limited availability of coastal land and water resources, effluent concerns, high production costs, and restricted growing seasons, all of which can be addressed through utilization of recirculating aquaculture systems. Although recirculating aquaculture systems offer greater controls over the rearing environment, these systems accumulate uneaten feed and waste generated by the fish, which if not removed become toxic and kill the fish. Ammonia removal efficiencies for two recirculating aquaculture system components, and particle size removal efficiencies for three other components, were determined under increasing feeding rates. The outcomes of these studies advance design criteria for developing technologies that will improve market competitiveness for domestic producers through maximizing environmental and production efficiencies, minimizing waste production, decreasing energy and regulatory costs, and optimizing product quality and safety. This accomplishment addresses Year 2 (2006) milestone, establish performance characteristics of recirculating aquaculture system components during low-salinity fish production and the following components of NP 106-Aquaculture: Aquaculture Production Systems; and Sustainability and Environmental Compatibility. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. ENTERGY USE, ENERGY EFFICIENCY AND WATER CONSUMPTION IN RECIRCULATING AQUACULTURE SYSTEMS DETERMINED: Energy use, energy efficiency, and water consumption were determined by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL for production scale recirculating systems. Energy is one of the major variable costs in an aquaculture production facility; therefore, a load analysis of electrical use helps target efficiency opportunities and minimizes operational costs. Complete energy load and daily water consumption analyses were conducted on four low-salinity production scale recirculating aquaculture systems consisting of four tanks each throughout a six-month growout production cycle of hybrid striped bass. The development of low-cost, energy efficient systems for sustainable marine aquaculture in low-salinity environments will increase job market opportunities for inland rural sectors and displaced farmers, increase crop diversity, and enhance food biosecurity. This accomplishment addresses Year 2 (2006) milestone, establish performance characteristics of recirculating aquaculture system components during low-salinity fish production and the following components of NP 106-Aquaculture: Aquaculture Production Systems; and Sustainability and Environmental Compatibility. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. POMPANO AND BLACK SEA BASS LARVAL DEVELOPMENT: Early life stages of Florida pompano and black sea bass were characterized via determination of growth and development by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL. Although limited knowledge exists regarding the timeframe of various physical and developmental changes that marine finfish larvae undergo, characterization of early life stages provides information which can be utilized to develop management protocols such as day of first feeding, appropriate size of live feed organisms to be offered, and time of complete metamorphosis. Larval Florida pompano and black sea bass were collected daily from culture tanks (from one day after hatching through complete metamorphosis) and microscopic images were captured and analyzed to determine parameters including length, yolk absorption rate, time of complete yolk absorption and first feeding, mouth size and development, and time of complete metamorphosis. This accomplishment will be used by fish hatchery managers to establish developmentally appropriate management methods for larval rearing of both species. This accomplishment addresses Year 2 (2006) milestone, characterize early development stages of Florida pompano and black sea bass, and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures DIETARY PROTEIN AND ENERGY FOR POMPANO: Optimal dietary protein and energy for rearing juvenile Florida pompano was determined by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL. Fish feed represents the highest operating cost at aquaculture facilities with protein accounting for the greatest percentage of this cost. Identifying optimal protein and energy levels for Florida pompano will decrease feed costs, reduce time to market, and increase feed efficiency while optimizing product quality, increasing food safety, and maximizing profits for fish culturists. Florida pompano were fed diets containing a wide range of protein and energy levels with fish receiving moderate levels of protein and high levels of energy performing as well or better than fish receiving higher levels of protein and less energy. This accomplishment will be used by fish feed manufacturers to make more efficient diets with lower protein levels creating a significant savings to feed manufacturers and fish culturists alike. This accomplishment addresses Year 2 (2006) milestone, develop reference research diets for Florida pompano, and the following component of NP 106-Aquaculture: Growth, Development, and Nutrition. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, Performance Measures 1.2.1, 1.2.2, and 1.2.5. NUTRIENT DIGESTIBILITY IN POMPANO: Scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL determined protein, energy, and other nutrient digestibility (availability) values for six protein sources that can serve as alternatives to fish meal for use in Florida pompano diets. High quality fish meal is the best source of protein for fish; however, replacement of fish meal with alternative protein sources will increase sustainability and profitability of the aquaculture industry. Three plant-based (soybean meal, soy protein concentrate, and corn gluten meal) and three waste by-product-based (poultry by-product meal, meat and bone meal, and distillers dried grains) feed ingredients were fed to Florida pompano and availability of protein, energy, and other nutrients were determined. This accomplishment will be used by fish feed manufacturers to substitute these novel and traditional feed ingredients for fish meal resulting in efficient, less expensive, and more environmentally responsible diets. This accomplishment addresses Year 2 (2006) milestone, develop reference research diets for Florida pompano, and the following component of NP 106-Aquaculture: Growth, Development, and Nutrition. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, Performance Measures 1.2.1, 1.2.2, and 1.2.5. BLACK SEA BASS BREEDING: A breeding and larviculture program for mass production of black sea bass juveniles was established by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL. A major factor limiting marine finfish aquaculture is the lack of techniques for sustained and reliable production of seedstock to supply grow-out operations. Captured black sea bass were induced to spawn, eggs were incubated and hatched, and the resulting larvae were reared through metamorphosis to produce juveniles. Results identified potential bottlenecks regarding broodstock acquisition and holding, while establishing baseline procedures for the successful mass production of juvenile black sea bass. Outcomes of this accomplishment will enhance U.S. competitiveness in marine fish culture through development of technologies for year-round seedstock production, enhanced biosecurity, and domesticated strains of broodstock to produce quality black sea bass seedstock. This accomplishment addresses Year 2 (2006) milestone, develop techniques and procedures for optimizing black sea bass spawning, and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. POMPANO AMMONIA AND NITRITE TOLERANCE: The tolerance of early life stages of Florida pompano to ammonia and nitrite was determined by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL. Ammonia and nitrite constitute the two major waste products encountered in recirculating aquaculture systems and knowledge of the levels detrimental to various life stages of fish is necessary for efficient management of culture systems. Eggs and larval Florida pompano of different ages (1, 8, and 16 days post-hatch) were subjected to short-term exposure (24 h) to varying concentrations of either ammonia or nitrite, and the tolerance of each life stage was determined. This accomplishment establishes the relative sensitivity of early developmental stages of pompano to ammonia and nitrite and this information can be used by hatchery managers to avoid detrimental concentrations of these aquaculture waste products. This accomplishment addresses Year 2 (2006) milestone, determine optimal temperature, salinity, light and density to enhance egg and larval survival, and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. POMPANO SENSITIVITY TO COPPER: The sensitivity of early developmental stages of Florida pompano exposed to copper was determined by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL. While copper is an effective treatment against various pathogens that can cause mass mortality of aquacultured fish, limited information exists with respect to the tolerance of early life stages of marine finfish. Eggs and larval Florida pompano of different ages (1, 8, and 16 days post-hatch), as well as post-metamorphic juveniles, were subjected to short-term exposure (24 h) to varying concentrations of copper and the tolerance of each life stage was determined. This accomplishment establishes the relative sensitivity of early developmental stages of pompano to copper allowing hatchery managers to effectively implement appropriate disease treatment protocols. This accomplishment addresses Year 2 (2006) milestone, determine optimal temperature, salinity, light and density to enhance egg and larval survival, and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. SALINITY EFFECTS ON LARVAL POMPANO: The effect of environmental salinity on early life stages of Florida pompano was determined by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL. Observations suggest Florida pompano are tolerant to a wide range of environmental salinities; however, limited information exists with respect to salinity effects on early developmental stages of pompano. Eggs and larval Florida pompano of different ages (1, 8, and 16 days post-hatch), as well as post-metamorphic juveniles, were subjected to short-term exposure (24 h) to varying salinity levels and the effects of salinity on each life stage was determined. This accomplishment establishes the range of salinities supporting acceptable egg buoyancy and hatching, as well as larval and juvenile survival and will increase the success rate and production efficiency of hatchery operators. This accomplishment addresses Year 2 (2006) milestone, determine optimal temperature, salinity, light and density to enhance egg and larval survival, and the following component of NP 106-Aquaculture: Reproduction and Early Development. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. MEASURING DISSOLVED CARBON DIOXIDE: Three methods for measuring dissolved carbon dioxide (CO2) in a marine recirculating aquaculture system were developed by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL. There are compounds and salts that can interfere with the determination of dissolved carbon dioxide concentrations in marine aquaculture systems; however, if the dissolved carbon dioxide gas can be released from the water it can be measured directly with a gas meter. Water at three salinities had three concentrations of dissolved carbon dioxide gas infused into them and the dissolved carbon dioxide gas in each combination of solutions was measured by three methods. This accomplishment indicates that removing carbon dioxide gas from water and measuring it directly with a portable hand-held gas analyzer provides a reliable and cost-effective method for fish culturists to measure CO2 concentrations in saltwater aquaculture environments. This accomplishment addresses Year 2 (2006) milestone, establish performance characteristics of recirculating aquaculture system components during low-salinity fish production, and the following components of NP 106-Aquaculture: Aquaculture Production Systems; and Sustainability and Environmental Compatibility. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. BIOFILTER NITRIFICATION: Rate of ammonia removal and performance curves for a fixed-film biofilter with floating bead media were determined by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL. Standard methods for determining and reporting biofilter performance for the purpose of comparing biofiltration efficiencies do not exist. Determining ammonia removal rates would therefore provide criteria to facilitate selection among the different types of biofilters. Four recirculating aquaculture systems with two fixed-film biofilters, each containing a different type of floating plastic bead media, received one of two different ammonia concentrations to evaluate ammonia removal at salinities ranging from 3 g/L to 15 g/L. This accomplishment provides aquaculturists, design engineers, and filter manufacturers the criteria needed to make well informed decisions for selecting biofilters for use over a wide range of salinities. This accomplishment addresses Year 2 (2006) milestone, establish performance characteristics of recirculating aquaculture system components during low-salinity fish production, and the following components of NP 106-Aquaculture: Aquaculture Production Systems; and Sustainability and Environmental Compatibility. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. AQUACULTURE INDUSTRY SURVEY: A survey of aquaculture industry experiences with different types of biological filtration was conducted by scientists at the ARS Sustainable Marine Aquaculture Project in Fort Pierce, FL. Recirculating technology for aquaculture relies almost exclusively on biofilters for ammonia removal and although many types of biofilters are commercially available, requirements for use in recirculating systems have not been systematically quantified. A survey was conducted canvassing over 400 people in the aquaculture industry including scientists and technicians at universities; private, public, and government research facilities; State and Federal hatchery managers; and aquaculture farmers for the purpose of quantifying filtration needs. This accomplishment provides biofilter manufacturers the demographics, requirements, and needs specific applications of biofilter users in recirculating aquaculture. This accomplishment addresses Year 2 (2006) milestone, establish performance characteristics of recirculating aquaculture system components during low-salinity fish production, and the following components of NP 106-Aquaculture: Aquaculture Production Systems; and Sustainability and Environmental Compatibility. The accomplishment also addresses ARS Strategic Plan Objective 1.2-Contribute to the efficiency of agricultural production systems and specifically, performance measures 1.2.1, 1.2.2, and 1.2.5. 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? Results of research studies and technologies developed were made available to customers and the general public through oral presentations (technical and non technical), poster presentations at local, state, national and international meetings, and scientific papers. 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). Weirich, C., Riley, K., Davis, M. 2005. Florida pompano: Induced reproduction via pelleted GnRHa and preliminary observations regarding larval production. Global Aquaculture Advocate. 8(3):75-77. Pfeiffer, T., Riley K. 2005. Filter performance study evaluates combined performance of bead, sand filters in RAS. Global Aquaculture Advocate. 8(4):39-42. Riley, K., Riche, M. 2005. Florida study achieves tank spawning of southern flounder on pelleted feed. Global Aquaculture Advocate. 8(5):52-53. Pfeiffer, T.J. 2005. Recirculating aquaculture system for marine finfish research at Harbor Branch Oceanographic Institution. Hatchery International. 6(6):20-23. Riley, K., Weirich, C.R. 2006. Florida's Harbor Branch commissions new marine recirculating system for spawning pompano and black sea bass. Hatchery International. 4:28. Webb, J., Wood, D., Smith, S., Pfeiffer, T. 2006. Practical insight on installation and use of a dual drain/swirl separator system. Fish Farming News. 13(2):18-19. Presentations: Weirich, C. March 2006. Marine finfish reproduction and larviculture techniques. Department of Aquaculture and Fisheries, University of Arkansas at Pine Bluff, Pine Bluff, Arkansas. Weirich, C. May 2006. Broodstock care and breeding techniques. Aquaculture Center for Training and Development, Harbor Branch Oceanographic Institution, Ft. Pierce, Florida. Riche, M. June 26, 2006. Integrating feeds and feeding management techniques into sustainable aquaculture strategies. Aquaculture Division, Harbor Branch Oceanographic Institution, Fort Pierce, Florida. Pfeiffer, T.J., Sumerfelt, S.T., Watten, B.J. July 21-23, 2006. Comparative performance of CO2 measuring methods: Marine recirculation system application. The Sixth International Conference on Recirculating Aquaculture, Roanoke, Virginia. Review Publications Weirich, C.R. 2006. Salinity effects on early life stages of Florida pompano Trachinotus carolinus [abstract]. Aquaculture America 2006 Book of Abstracts. p. 348. Weirich, C.R., Riche, M.A. 2006. Acute tolerance of juvenile Florida pompano, Trachinotus carolinus L. to ammonia and nitrite at various salinities. Aquaculture Research. 37:855-861. Riley, K.L., Weirich, C.R., Webb, J.B. 2006. Design and operation of a marine recirculating system for controlled spawning of Florida pompano and black sea bass [abstract]. Aquaculture America 2006 Book of Abstracts. p. 248. Riche, M.A., Pfeiffer, T.J., Garcia, J. 2006. Evaluation of a sodium hydroxymethane-sulfonate product for reducing total ammonia nitrogen in a small-scale rotifer batch culture system. North American Journal of Aquaculture. 68:199-205. Rawles, S.D., Riche, M.A., Gaylord, T.G., Webb, J., Freeman, D.W., Davis, M. 2006. Evaluation of poultry by-product meal in commercial diets for hybrid striped bass (Morone chrysops x M. saxatilis) in recirculated tank production. Aquaculture. 259:377-389. Weirich, C.R., O'Neal, C.C., Belhadjali, K. 2005. Growth, body composition, and survival of channel catfish ictalurus punctatus fry fed hatchery diets supplemented with krill meal. Journal of Applied Aquaculture. 17(3): 21-35. Reigh, R.C., Williams, M.B., Gillespie, J.M., Weirich, C.R. 2006. Preliminary observations on the cost of producing fingerlings from channel catfish stocked as sac fry or as hatchery-fed swim-up fry. Journal of Applied Aquaculture. 18(3):75-84. Pfeiffer, T.J., Malone, R. 2006. Nitrification performance of a propeller-washed bead clarifier supporting a fluidized sand biofilter in a recirculating warmwater fish system. Aquacultural Engineering. 31(1):80-92.