Progress 10/01/06 to 09/30/07
Outputs
Progress Report Objectives (from AD-416) To vertically integrate (from raw materials to feeds to food products) the science and technology of producing Pacific white shrimp, and the model tropical fish Pacific threadfin and longfin amberjack. The project focuses on optimizing existing technologies for shrimp growout, developing new technologies for threadfin (including larvae) and amberjack growout, and use of agricultural by-products and reduced fish meal in feeds for these species. These technologies have significant potential for expansion to tropical areas of the U.S. Approach (from AD-416) This project advances the science and technology of developing feeds for optimum nutrition of cultured shrimp and finfish, using a vertically integrated approach. Its three distinct but interdependent objectives are: develop commercially viable feeds and processing methods using tropical agricultural by-products; establish nutritional and biochemical strategies for maximizing larval and growout productivity and reducing fish meal in feeds for tropical species; and establish product quality of species reared on the developed feeds. Successful completion of this project will fill a knowledge gap in the science and technology of culturing the tropical species Pacific white shrimp, Pacific threadfin, and longfin amberjack. The resultant technology will have a significant impact on aquaculture in Hawaii, other Pacific Basin islands, and tropical regions of the U.S. Expansion of domestic aquaculture production could help offset part of the national trade deficit in fisheries products, and promote a sustainable aquaculture industry in the U.S. Formerly 5320-31000-006-00D (1/2005) Significant Activities that Support Special Target Populations This report serves to document research conducted under a Grant agreement between ARS and the Oceanic Institute. Additional details can be found in the report for the parent project 5320-31000-007-00D Tropical Aquaculture: Develop Feeds for Optimum Nutrition of Cultured Shrimp and Finfish. Progress reported in Question 4 under Accomplishments. Accomplishments Inclusion of agricultural by-products in diets for Pacific white shrimp, Litopenaeus vannamei: This work addressed two problems: developing cost-effective shrimp feeds that use locally available ingredients, and finding new uses for by- products that would otherwise require costly disposal. A shrimp growth trial was conducted by scientist at the U.S. PBARC, Hilo HI,at the Oceanic Institute in Waimanalo, Hawaii, using experimental diets containing one of several agricultural by-products: whole papaya (10%, freeze-dried), papaya flesh (5%, freeze-dried), and papaya seeds (10%, freeze-dried); wheat mill run (44.8%); and okara (pulp left after soybeans are pressed) at two inclusion levels (10 and 20% in partial replacement of fishmeal); a fishmeal-based diet, which contained none of these by-products, was used as a control. This trial was conducted in an outdoor, zero-water exchange system that permitted the development of high levels of floc, which has been shown to provide a portion of the nutrient needs of shrimp. After eight weeks of feeding, the experimental diets containing by-products performed as well as the control diet in terms of shrimp final weight, growth, and survival; by comparison, shrimp in another treatment being fed a commercial diet had lower growth rate and final weight. The results of this trial can be useful in manufacturing cost-effective shrimp feeds, while providing increased opportunity for the utilization of agricultural by-products. (NP 106 Action Plan Component4: Growth, Development, and Nutrition. Problem being addressed: C.Sustainable Sources of Nutrients). Lysine requirement of Pacific threadfin: The requirement of Pacific threadfin for lysine was determined in a study using a control diet formulated to contain lysine at a level considerably below known requirements for other fish. The amino acid requirements of threadfin are largely unknown, and this prevents optimum diet formulation for this species. A growth trial was conducted by researchers in Hilo, HI, in which diets containing graded levels of crystalline lysine supplemented in replacement of glutamic acid (an amino acid generally considered to be non-essential) were fed to juvenile threadfin for six weeks. The growth and feed efficiency data were compared with the dietary lysine levels to establish the requirement (i.e. the minimum dietary lysine level needed to maximize growth and feed efficiency), which was found to be 1.36% of the diet, corresponding to 3.51% of dietary protein. This information can be used to formulate improved diets for this important food fish species. (NP 106 Action Plan Component4: Growth, Development, and Nutrition. Problem being addressed: D.Nutrient Use and Feed Evaluation). A practical methodology for testing feeding stimulants on shrimp: A method for evaluating ingredients as feeding stimulants for shrimp was improved by modifying the experimental test system, feed delivery, and consumption estimation, and applying new statistical methods for data evaluation. This method was used to examine the effect on voluntary consumption by shrimp of diets containing hydrolysates made from fish processing by-products. It was found that none of the hydrolysates were repellants (reduced feed consumption relative to bland control), but only two were found to be attractants (increased feed consumption relative to control). This improved methodology provides an efficient, repeatable means of evaluating ingredients for shrimp feeds, and is the subject of a manuscript that has been submitted for publication in a peer-reviewed journal. Collaborative research on this subject with Dr. Addison Lawrence of Texas A&M University is also anticipated. (NP 106 Action Plan Component4: Growth, Development, and Nutrition. Problem being addressed: D.Nutrient Use and Feed Evaluation). Development of a standardized shrimp feed for processing research: A standardized feed for shrimp was profiled and manufactured under the following National Institutes of Health Standards (NIH) specifications: NIH STD 1 � Animal Feed Processing and Milling Sanitation Standard; and NIH STD 5 � Nutrient and Chemical Contaminant Analysis of Laboratory Animal Diets. In collaborative work with Dr. Addison Lawrence of Texas A&M University and Zeigler Brothers feed company, an open shrimp feed formulation was produced, with defined ingredients and a defined nutrient profile, which would be able to measure the performance of animals, culture systems, processing methods, and cost against all feeds tested for shrimp. The standard feed was made at Zeigler Brothers, and when tested at TAMU and OI research labs, showed poor performance; in addition to the poor performance, an FDA-unapproved contaminant, melamine, was discovered in the binder used to make the feed. An ingredient particle size trial was also run using the standardized feed, with the same results. Analyses are currently underway with Zeigler, Dr. Lawrence, and OI to determine why the standardized feed performed so poorly. (NP 106 Action Plan Component: Growth, Development, and Nutrition. Problem being addressed: Nutrient Use and Feed Evaluation). Determined fatty acid profiles of longfin amberjack fillet: Scientist at the U.S. PBARC Hilo, HI in collaboration with an offshore sea-cage producer of longfin amberjack identified a need for information on the nutritional requirements of this species. Analysis of the fatty acid content of cultured longfin amberjack fillets revealed levels of highly unsaturated fatty acids comparable to those found in farmed salmon. This knowledge provides insight into the nutritional needs of the animal, which can lead to more nutritionally-efficient feeds for a new species in the aquaculture industry. (NP 106 Action Plan Component4: Growth, Development, and Nutrition. D.Problem being addressed: Nutrient Use and Feed Evaluation). Effect of floc bioactive compounds on shrimp growth: Although it is known that culturing shrimp in the presence of floc leads to improved growth, it is not known which components of floc are responsible for this. As a first step in isolating and identifying bioactive compounds in floc that enhance shrimp growth, various floc solvent extract fractions, each potentially containing different classes of compounds, were incorporated into a complete formulated diet (control diet) for shrimp. Scientists in Hilo, HI produced seven experimental diets with inclusions of either whole floc (intact floc and ground floc), or one of the following solvent fractions of floc: polar (Fraction A), medium-polar (Fraction B), non-polar (Fraction C), and the insoluble residue (Fraction D), individually and recombined (A+B+C+D). An indoor growth trial using these diets demonstrated that shrimp fed the diet containing floc Fraction B (medium-polar extract containing photopigments, phytosterols, polyphenols, bromophenols, glucosamine, and fat soluble vitamins) achieved significantly higher weekly growth than those fed the control diet. Knowledge of the beneficial bioactive components in the shrimp floc could lead to improved shrimp feed formulations that support optimal growth. (NP 106 Action Plan Component: Growth, Development, and Nutrition. D.Problem being addressed: Nutrient Use and Feed Evaluation). Evaluated medium-polar bioactive compounds from floc in formulated shrimp diets: In a continuation of the research to identify which components of the floc are responsible for enhanced shrimp growth, experimental diets were produced by ARS scientists in Hilo, HI with commercially-available, potentially bioactive compounds incorporated into a complete formulated diet (control diet) for shrimp. Eight experimental diets were produced with selected groups of compound inclusions in the fat top-coating of the diet: carotenoid pigments, lutein, brown seaweed, astaxanthin, phytosterols, bromophenols, glucosamine, and a combination of all the compounds; a commercially feed (40% crude protein) was also included in the trial. An eight-week indoor growth trial using these diets did not produce significantly enhanced growth from the control. However, when compared to the commercially available feed, all the formulated diets yielded significantly better shrimp growth (P<0.05) than the commercial feed. A follow-up growth trial with inclusions of graded levels of selected bioactive compounds (astaxanthin and bromophenol), as well as direct inclusion of the compounds rather than top-coating, may produce significant growth differences. Knowledge of the beneficial bioactive components in the shrimp floc could lead to improved shrimp feed formulations that support optimal growth. (NP 106 Action Plan Component: Growth, Development, and Nutrition. Problem being addressed: D.Nutrient Use and Feed Evaluation). Nutritional contribution of microbes in super-intensive shrimp culture systems: Bulk tissue and amino acid-specific stable isotope methods were used to better understand the role of the microbial community present in high intensity, zero-exchange shrimp culture systems, in the nutrition of animals grown in those systems. Shrimp and total particulate organic matter samples were collected weekly for 14 weeks from triplicate light and dark systems; preliminary bulk tissue data suggest that shrimp did not gain significant carbon or nitrogen from the microbial community in systems exposed to light, but in dark systems, shrimp could have been ingesting the microbial community. However, because the isotopic signature of the bulk microbial community nitrogen was not significantly different from that of the feed in the dark, the two potential sources of nitrogen could not be distinguished isotopically. Amino acid-specific isotope analysis of shrimp and microbial community samples will allow us to elucidate the latter; these analyses are almost complete and our early results confirm our bulk tissue analyses. The identification of alternate, naturally-occurring nutrition sources available to shrimp in super-intensive production systems is important for an understanding of the potential for increased system efficiency, shrimp growth rate, and feed conversion ratio.� (NP 106 Action Plan Component4: Growth, Development, and Nutrition. Problem being addressed: D.Nutrient Use and Feed Evaluation). Improved hatchery hygiene and nutrition for larval threadfin Hatchery modifications were initiated to improve hatchery hygiene, lighting, and temperature control in larval threadfin rearing tanks, and were followed by several large-scale hatchery runs to examine the effects of background algae (live vs. paste), live feed density, and degassing on larval survival. Five production runs were initiated, with two successfully yielding larvae to day 25, and three crashing soon after initiation of rotifer feeding. Results were as follows: Trial 1 yielded 75,000 post-larvae/52,000 fingerlings with slightly improved survival in tanks using algae paste as background algae, and rotifer feed over those reared using live algae; Trial 2 crashed; Trial 3 yielded 209,000 post- larvae/105,000 fingerlings and demonstrated increased survival (17%) using higher live feed densities and lower water exchange compared with controls (12%); Trials 4 and 5 unsuccessfully examined the effect of degassing larval rearing tank water, with both trials crashing during the critical early larval feeding period, with lower total gas pressure (from ~111% to ~103%) showing no beneficial effects on hatchery performance. Despite these mixed results, the trials assisted industry by providing practical hatchery training and fingerlings for industry growout, and by ruling out algae background, rotifer diet, rotifer density, and total gas pressure as possible causes of the early hatchery mortality. The unpredictable and intermittent nature of the hatchery crashes and lack of correlation with larval rearing method suggests a need to improve hatchery biosecurity, and to study potential vectors for pathogen entry into the larval rearing system, including egg supply, water, air, algae, and rotifers. (NP 106 Action Plan Component: Growth, Development, and Nutrition. Problem being addressed: Regulating Feed Intake. NP 106 Action Plan Component3: Reproduction and Early Development. Problem being addressed: E.Early Life Stage Development and Survival). Effects of broodstock nutrition and/or photoperiod on threadfin egg quality Although the broodstock were very slow to acclimate to formulated test diets, both the commercial diet (Vitalis salmon broodstock) and the OI formulated and manufactured diet, successfully maintained spawning broodstock and yielded viable fertilized eggs for use in the hatchery. Although the test diets did not have any significant effects on hatch rate, larval growth, or early hatchery survival, poor early hatchery performance persisted under all dietary regimens; in addition, the greater iodine availability through formulated diets, especially the high- iodine OI diet, increased egg thyroid hormone levels from hypothyroid status relative to broodstock fed a raw diet, and resulted in reduced rates of goiter formation and lower rates of broodstock mortality. The effect of broodstock diet on egg biochemistry is awaiting completion of biochemical analyses. Although additional work is needed to determine the optimal diet composition for improved egg quality and hatchery performance, both the commercial diet, and in particular, the Oceanic Institute diet, improved broodstock survival. The results of these trials will assist industry by providing a biosecure formulated diet capable of enhancing broodstock performance and viability. (NP 106 Action Plan Component4: Growth, Development, and Nutrition. Problem being addressed: E.Interactions Affecting Reproduction). Publications produced are as follows: Decamp, O., Conquest, L, Cody, J., Forster, I.P., Tacon, A.G.J. 2007. Effect of shrimp stocking density on size-fractionated phytoplankton and ecological groups of ciliated protozoa within zero-water exchange culture systems. Journal of the World Aquaculture Society 38(3):395-406. Forster, I.P., Dominy, W.G. 2006. Efficacy of three methionine sources in diets for Pacific white shrimp, Litopenaeus vannamei. Journal of the World Aquaculture Society 37:474-480. Moss, S.M., Forster, I.P., Tacon, A.G.J. 2006. Sparing effects of pond water on vitamins in shrimp diets. Aquaculture 258 (1-4):388-395. Verner-Jeffreys, D.W., Nakamura, I., Shields, R.J. 2007. Surface disinfection of Pacific threadfin, Polydactylus sexfilis and amberjack, Seriola rivoliana, eggs. Aquaculture Research 38:605-612. Technology Transfer Number of Non-Peer Reviewed Presentations and Proceedings: 6 Number of Newspaper Articles,Presentations for NonScience Audiences: 1
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Progress 10/01/05 to 09/30/06
Outputs
Progress 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? The United States had an estimated annual trade deficit of $8 billion in edible and non-edible fisheries products in 2005, including $3.4 billion in shrimp imports. This trade deficit is the largest of any agricultural commodity and second only to petroleum for any natural resources product. Expansion of domestic aquaculture production could offset part of this trade deficit and help establish a globally competitive, sustainable (i.e. environmentally non-degrading, technically appropriate, economically viable, and socially acceptable) aquaculture industry in the U.S. Despite considerable scientific advances, optimum feeds, especially those with low or no fish meal, for maximum productivity of Pacific white shrimp (Litopenaeus vannamei) under sustainable culture conditions have not yet been
established. Knowledge of the impact of such feeds and the culture environment on product quality and safety of cultured shrimp is limited. A great need exists to address the above problems in support of the U.S. shrimp industry. In Hawaii every year, disposal of numerous by-products that have potential use in the aquatic feed industry (e.g. meat and bone meal, wheat mill run, coffee bean pulp, macadamia nut presscake, molasses) results in economic loss and potential environmental impacts. Identification and nutrient profiling of potential tropical by-products will permit expansion of aquatic feed research to include the use of these potential ingredients. Pacific threadfin (Polydactylus sexfilis), known locally as moi, and longfin amberjack (Seriola rivoliana), known locally as kahala, are both fast-growing, good-tasting tropical marine carnivorous fish. These fish have been studied at the Oceanic Institute (OI) for the past several years and have shown good market potential.
Threadfin and amberjack culture are growing industries in Hawaii and other tropical regions, and there is a need for research on nutrient requirements and feed development, especially those using fish meal alternatives. The effects of feed type, feeding strategy, and culture environment on flesh quality and growth of these fish require further investigation. Research is needed to ensure that the high consumer preference for these animals is maintained under intensive, sustainable culture conditions. Our proposed project takes an integrated approach in addressing the issue of providing the U.S. with healthful, desirable seafood. The potential benefits from attaining these objectives include: an enhanced understanding of vertically-integrated science and technology for production of shrimp, and the model tropical fish, threadfin and amberjack; development of improved feeds with low or no fish meal for tropical species. Improved use of tropical agricultural by-products, feeding
technologies, culture management methodology, environmental compatibility, and growth of the U.S. shrimp and tropical marine finfish aquaculture industry; an expanded range of high-quality aquacultured products which will benefit U.S. consumers and food processors, and an increased consumer confidence in the safety of aquatic foods through quality assurance. Anticipated products of the research include lower-cost, low environmental impact aquaculture feeds with low or no fish meal for use in intensive culture systems, using improved growout feed formulations that employ commercially available feed ingredients and selected by- products of the agriculture industry; reduction in reliance of tropical regions on imported ingredients for aquaculture feed production, improved techniques for assessing advanced physical, chemical, and nutritional quality of feed ingredients and finished feeds, development of feeds and feeding practices for maximum productivity of shrimp and two high-value
marine finfish, and improved product quality of cultured shrimp, threadfin, and amberjack. The resulting vertically-integrated science and technology can serve as a contribution to the U.S. aquaculture industry. Primary customers of this research are U.S. aquaculture farmers, aquafeed manufacturers, feed ingredient suppliers (including those that generate agriculture by-products), and aquatic food processors who are able to directly apply the results of this research. Equipment manufacturers (feed processing; food quality evaluation) and seafood consumers will benefit from improved technology and increased availability of high-quality, affordable aquacultured products. This project is primarily devoted to addressing the growth, development, and nutrition component of NP 106 Aquaculture. It also supports the NP 106 components involving aquaculture production systems, and quality, safety, and variety of aquacultured products for consumers. The objectives of this project are in
alignment with the NP 106 Mission Statement: Conduct high quality, relevant, basic and applied aquaculture research to improve the efficiency, profitability, and sustainability of United States aquaculture, and reduce dependence on imported seafood and threatened ocean fisheries. 2. List by year the currently approved milestones (indicators of research progress) OBJECTIVE I: DEVELOP COMMERCIALLY VIABLE FEEDS AND PROCESSING METHODS USING TROPICAL AGRICULTURAL BY-PRODUCTS Sub-objective 1. Tropical Agricultural By-Products and Ingredients Characterization Sub-objective 2. Feed Processing Sub-objective 3. Feed Development and Characterization OBJECTIVE II: ESTABLISH NUTRITIONAL AND BIOCHEMICAL STRATEGIES FOR MAXIMIZING LARVAL AND GROWOUT PRODUCTIVITY AND REDUCING FISH MEAL IN FEEDS FOR TROPICAL SPECIES Sub-objective 4. Nutritional Biochemistry and Microbiology of Species and Culture Systems Sub-objective 5. Nutrition for Maximum Productivity and Use of Alternatives to Fish
Meal OBJECTIVE III: ESTABLISH PRODUCT QUALITY OF SPECIES REARED ON DEVELOPED FEEDS Sub-objective 6. Product Quality of Cultured Species Reared on Developed Feeds Year 1 (2005) Tropical Agricultural By-Products and Ingredients Characterization Completed by-product identification Feed Processing Completed pellet moisture variation analyses Feed Development and Characterization Protocol for attractant testing completed Nutritional Biochemistry and Microbiology of Species and Culture Systems Shrimp cultured in raceways to produce and collect floc Pacific Threadfin (Moi) Hatchery Trials 1 Through 4: 1. Examine the role and effects of background algae during the rotifer feeding phase of threadfin larval development and its impact on hatchery survival and early fingerling growout 2. Examine the effects of rotifer enrichment protocols on threadfin larval development and hatchery survival 3. Compare threadfin production in the OI finfish hatchery with that in the newly opened
Pilot Production hatchery facility 4. Examine the effect of rotifer density on threadfin production output in the newer Pilot Production facility Nutrition for Maximum Productivity and Use of Alternatives to Fish Meal Reduction by half of fish meal from shrimp feeds in zero-water exchange systems Product Quality of Cultured Species Reared on Developed Feeds Dietary lipid and cholesterol effect on shrimp quality determined Year 2 (2006) Tropical Agricultural By-Products and Ingredients Characterization Completed by-product ingredient characterization Feed Processing Completed water stability and pellet durability (PDI) analyses Feed Development and Characterization Commercially viable diet formulated for shrimp Nutritional Biochemistry and Microbiology of Species and Culture Systems Fractionation of floc in aqueous and solvent phase to obtain fractions (A, B, C, etc.) complete Fractions A, B, C, etc. tested in shrimp feeds Nutrition for Maximum Productivity and Use of
Alternatives to Fish Meal Methionine and lysine requirement studies with threadfin completed Develop a formulated broodstock diet for Pacific threadfin Determine effect of diet and enrichments on nutritional biochemistry of s- type rotifers Determine biochemical changes related to diet during larval development of threadfin Develop weaning and nursery diets for threadfin Initial determination of digestive enzyme ontogeny regulation in first feeding larval threadfin Product Quality of Cultured Species Reared on Developed Feeds Methionine and lysine effect on fish quality evaluated Year 3 (2007) Tropical Agricultural By-Products and Ingredients Characterization Diet formulation with by-product ingredients completed. Feed Processing By-product processing protocols completed. Lab-scale processing modifications completed. Feed Development and Characterization Physical and chemical analysis of diets completed. Optimal diet for shrimp determined. Nutritional Biochemistry and Microbiology of
Species and Culture Systems Chromatographic separation to obtain fractions a, b, c, etc. completed Nutrition for Maximum Productivity and Use of Alternatives to Fish Meal Fatty acid requirements for fish evaluated Product Quality of Cultured Species Reared on Developed Feeds Effect of exercise and feed with required lipid on fish quality completed. Effect of dietary fatty acid on fish quality completed. Effect of best attractants and optimum feeding strategies on fish and fish quality determined. Year 4 (2008) Tropical Agricultural By-Products and Ingredients Characterization (Specific milestones not anticipated at this stage for this sub-objective. ) Feed Processing (Specific milestones not anticipated at this stage for this sub-objective. ) Feed Development and Characterization Shrimp diet recommended to industry. Commercially viable diet formulated for fish Attractants identified Nutritional Biochemistry and Microbiology of Species and Culture Systems Fractions a, b, c, etc.
tested in shrimp feeds Structural elucidation of fractions a, b, c, etc. competed Feeding trials to establish sparing effect of sulfur amino acids completed Nutrition for Maximum Productivity and Use of Alternatives to Fish Meal Lysine requirement studies completed (amberjack) Protein:energy ratios in diets for fish optimized Fish meal and oil reduced by half in fish diets Product Quality of Cultured Species Reared on Developed Feeds Optimum protein:energy ratios effect on fish quality determined. Reduced fish meal / fish oil effect on fish quality completed. Year 5 (2009) Tropical Agricultural By-Products and Ingredients Characterization Optimal by-product inclusion in diet determined Feed Processing Recommend diet/process to industry Feed Development and Characterization Optimal diet for fish determined Nutritional Biochemistry and Microbiology of Species and Culture Systems Pilot-scale evaluation of shrimp growth-promoting compounds completed Nutrition for Maximum Productivity and
Use of Alternatives to Fish Meal Effects of exercise and lipid level on fish established Fish meal and oil reduced by 2/3 in fish diets Product Quality of Cultured Species Reared on Developed Feeds Correlation of fish quality with protein/energy ratios and amino acids completed Correlation of fish quality with dietary lipid, fatty acids, and exercise completed Quality of shrimp and fish reared on commercially viable feeds established 4a List the single most significant research accomplishment during FY 2006. All of the work described below was conducted at the Oceanic Institute in Waimanalo, Hawaii. Determined effects of diet and enrichment on nutritional biochemistry of s-type rotifers: Biochemical analyses of the nutritional profile of rotifers confirmed that their fatty acid composition, and therefore their nutritional value for larval threadfin (Polydactylus sexfilis), could be enhanced through an optimum combination of rotifer feeding and enrichment. Rotifers are deficient in
some fatty acids, such as the beneficial omega- 3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are essential for neural development, growth, and energy in larval fish. Five different feed treatments were tested to evaluate the most nutritious enrichment protocol for rotifers destined to be used as live feeds for larval threadfin. The rotifers raised on readily available algae (Nannochloropsis sp.), and subsequently enriched with a commercial supplement consisting of spray-dried Schyzochitrium algae, had the highest levels of EPA and DHA. This information can help identify the best and most cost-effective feeding and enrichment regimes to use in producing highly nutritious live feeds for larval threadfin. This accomplishment is aligned with the NP 106 Action Plan component on Growth, Development, and Nutrition, and addresses the problem of Nutrient Use and Feed Evaluation. 4b List other significant research accomplishment(s), if any. Inclusion of
agricultural by-products in diets for Pacific white shrimp: Several by-products of Hawaii agriculture were found to be suitable for inclusion as ingredients in feeds for Pacific white shrimp, Litopenaeus vannamei. This work addressed two problems: developing cost-effective shrimp feeds that use locally available ingredients, and finding new uses for by-products that would otherwise require costly disposal. A shrimp growth trial was conducted using experimental diets containing one of several agricultural by-products: meat & bone meal at four inclusion levels; spent fruit-fly media; and macadamia nut grit (nutcake remaining after extraction of macadamia oil) at two inclusion levels; a fishmeal- based diet, which contained none of these by-products, was used as a control. This trial was conducted in an outdoor, zero-water exchange system that permitted the development of high levels of floc, which has been shown to provide a portion of the nutrient needs of shrimp. After eight weeks of
feeding, there were no significant differences in final weight, growth, or survival between shrimp fed the experimental diets and those fed the control. The results of this trial can be useful in manufacturing cost-effective shrimp feeds, while providing increased opportunity for the utilization of agricultural by-products. This accomplishment is aligned with the NP 106 Action Plan component on Growth, Development, and Nutrition, and addresses the problem of Sustainable Sources of Nutrients. Improved nursery diet for moi (Pacific threadfin): Initial work to develop an improved nursery feed for threadfin (Polydactylus sexfilis) indicated that higher dietary protein levels led to increased threadfin nursery production (total final weight of fish) and feed efficiency. Because there are no feeds specifically developed for the nursery phase of this species, it has been necessary to use a commercial feed formulated for other species, which may not adequately address the nutritional needs of
juvenile threadfin. Juvenile fish were weaned from a live feed regimen onto a series of dry diets containing 43, 47, 51, or 55% crude protein. Although mortality was too high for the findings to be definitive, it was found that production (survival x final weight) was positively correlated with dietary protein level, with the highest production found in the 55% protein treatment. These findings, if corroborated by a new trial (with steps taken to reduce mortality), could enable development of more effective feeds for juvenile threadfin, which will promote better growth and a shorter time in the nursery. This accomplishment is aligned with the NP 106 Action Plan component on Growth, Development, and Nutrition, and addresses the problem of Tissue Growth and Development. Floc fractions tested in shrimp feeds: Using methods developed in 2005 research, potential bioactive components were extracted from shrimp pond floc and incorporated into formulated diets for Pacific white shrimp,
Litopenaeus vannamei. Although the supplemental nutrition from floc is beneficial in shrimp ponds, production and management of the desired floc community is difficult to achieve and produces variable results. Identifying theNBbeneficial components of pond floc and incorporating them into formulated feeds would enable the shrimp industry to utilize these components without extensive pond management. A shrimp growout trial using feeds containing floc components is underway, and the biochemical composition (proximate, amino acids, fatty acids, carotenoids, fat-soluble vitamins) of whole floc samples and 11 formulated feeds containing various extracted fractions has also been determined. Knowledge of the beneficial bioactive components in the shrimp floc could lead to improvements in shrimp feed formulations that result in optimal growth. In addition, certain floc components (chlorophyll and carotenoids) could be used to estimate the composition of the phytoplankton community present in
shrimp ponds. This accomplishment is aligned with the NP 106 Action Plan component on Growth, Development, and Nutrition, and addresses the problem of Nutrient Use and Feed Evaluation. Effect of finishing dietNBon product quality of moi (Pacific threadfin): A preliminary determination was made on the effect of lipid content in a finishing diet on fillet yield, composition, and sensory quality of harvested threadfin. This study was undertaken because no data are currently available on the effect of dietary lipid levels on product quality of this species. Juvenile threadfin were grown close to market size, and then fed with a finishing diet containing 43% protein, and either 13% or 24% lipid, for 14 weeks before harvest to enhance product quality. Results have indicated that the 24% lipid finishing diet did not lower the fillet yield of threadfin; data analysis to determine whether there were any differences in fillet composition and sensory quality is in progress. The results of this
study will provide baseline data that can be used in formulating cost-effective finishing diets toNBenhance and ensureNBfish product quality. This accomplishment is aligned with the NP 106 Action Plan components on Growth, Development, and Nutrition; and Quality, Variety, and Safety of Aquaculture Products. The problems it addresses include Nutrient Use and Feed Evaluation; and Predicting Product Quality or Defects. 5. Describe the major accomplishments to date and their predicted or actual impact. Two academic prototype shrimp feeds (one for indoor culture, one for outdoor) were developed which provide excellent performance, often outperforming commercial feeds. The commercially viable versions of these feeds are being formulated and tested, after which they will be moved to pilot scale for commercial applications. The problems addressed by this accomplishment include the traditional use in academic feed formulations of ingredients that are costly and not available year-round or in
large quantities, or contain too many ingredients in the formulation, which increases the time required for manufacturing, thereby increasing the inventory, need for storage space, number of purchases, and number of tracking and accounting entries and payments. The objective is to simplify the formulation and use common feed ingredients currently available to the feed industry at the minimum cost. (NP 106 Action Plan Component: Growth, Development, and Nutrition. Problem being addressed: Sustainable Sources of Nutrients; Nutrient Use and Feed Evaluation). Different forms of methionine (including L-methionine, DL-methionine and HMTBA [(2-hydroxy-4-methylthio) butanoic acid]) were shown to be equal in their ability to meet the needs for dietary methionine of shrimp grown in low-water exchange pond-like conditions. Use of HMTBA could enable formulation of commercial shrimp feeds with higher levels of soybean meal in replacement of fish meal, while meeting the nutritional needs of the
animal. (NP 106 Action Plan Component: Growth, Development, and Nutrition. Problem being addressed: Nutrient Use and Feed Evaluation). In ongoing research toNBisolate bioactive compounds in the microbial community (floc) of shrimp culture systemsNBthat are beneficial to shrimp nutrition, wet floc was collected from shrimp raceways, freeze-dried, and fractionated into aqueous and solvent phases. Analysis of the components indicated a composition of 12% dry matter, 32% crude protein, and 2% crude fat. In the course of this research, new methods were developed to fractionate the floc and isolate various components via HPLC and GC analysis, including carotenoid pigments, bioactive sterols, peptides and phenolic antioxidants, amino acids, fatty acids, and vitamins. This research has the potential to assist feed manufacturers in producing improved feeds for better shrimp performance. NP 106 Action Plan Component: Growth, Development, and Nutrition. Problem being addressed: Nutrient Use
and Feed Evaluation). An experiment in which shrimp were fed a diet containing a veryNBlow level of cholesterol and a highNBlevel of docosahexaenoic acid (DHA) resulted in a lower level of cholesterol and a higher level of DHA in theNBtail muscle tissue of market size shrimp, compared to shrimp fed a commercial control diet.NBResults clearly showed that shrimp product quality can be improved (i.e., positive quality attributes enhanced, and negative attributes reduced) by substantially manipulating the ingredients incorporated into the diet. (NP 106 Action Plan Components: Growth, Development, and Nutrition; Quality, Safety, and Variety of Aquaculture Products. Problems being addressed: Nutrient Use and Feed Evaluation; Predicting Product Quality or Defects). Biochemical analyses of the nutritional profile of rotifers confirmed that their fatty acid composition, and therefore their nutritional value for larval threadfin (Polydactylus sexfilis), could be enhanced through an optimum
combination of rotifer feeding and enrichment. Rotifers raised on readily available algae (Nannochloropsis sp.), and subsequently enriched with a commercial supplement consisting of spray-dried Schyzochitrium algae, had the highest levels of the beneficial omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are essential for neural development, growth, and energy in larval fish. This information can help identify the best and most cost-effective feeding and enrichment regimes to use in producing highly nutritious live feeds for larval threadfin. (NP 106 Action Plan Component: Growth, Development, and Nutrition. Problem being addressed: Nutrient Use and Feed Evaluation). Several by-products of Hawaii agriculture were found to be suitable for inclusion as ingredients in feeds for Pacific white shrimp, Litopenaeus vannamei. This work addressed two problems: developing cost-effective shrimp feeds that use locally available ingredients, and finding new
uses for by-products that would otherwise require costly disposal. A shrimp growth trial was conducted using experimental diets containing one of several agricultural by-products: meat & bone meal at four inclusion levels; spent fruit-fly media; and macadamia nut grit (nutcake remaining after extraction of macadamia oil) at two inclusion levels; a fishmeal- based diet, which contained none of these by-products, was used as a control. After eight weeks of feeding, there were no significant differences in final weight, growth, or survival between shrimp fed the experimental diets and those fed the control. The results of this trial can be useful in manufacturing cost-effective shrimp feeds, while providing increased opportunity for the utilization of agricultural by- products. (NP 106 Action Plan Component: Growth, Development, and Nutrition. Problem being addressed: Sustainable Sources of Nutrients). Note: In the accomplishments listed above, the potential beneficiaries include U.S.
aquaculture farmers, aquafeed manufacturers, feed ingredient suppliers (including those that generate agriculture by-products), and aquatic food processors who are able to directly apply the results of this research. Equipment manufacturers (feed processing; food quality evaluation) and seafood consumers, will also benefit from improved technology and increased availability of high-quality, affordable aquacultured products. 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? A presentation involving an overview of the Oceanic Institute, including its goals, departments, and achievements, was given at the 22nd Alltech International Feed Industry Symposium in Lexington, Kentucky. Over 1,500 people from 70 countries attended the symposium by invitation
only. Special Sessions focused on pigs, poultry, ruminants, aquaculture, pets, horses and foods. The audience consisted primarily of feed industry professionals and technical specialists, some academic, but mostly applied researchers, nutritionists, and veterinarians. While the overall program emphasized food animal products, the relationship to human nutrition and health issues was an integral part of the symposium. A series of lectures on the use of terrestrial animal by-products in aquatic feeds was given by one of the projects research scientists during a trip through China, Indonesia, and Vietnam. The lectures were sponsored by a U.S. national trade association that is active in the area of rendering. In each venue, over 50 people representing the feed manufacturing and aquaculture industries attended. The purpose of these lectures was to present information about the use of animal by-products, especially meat and bone meal and poultry meals, in support of marketing efforts by
the trade association to promote U.S. by-products in these countries. Most of the information presented was based on research conducted at the Oceanic Institute. Following completion of the new Product Innovation Laboratory last year, this facility is now being used to provide the Oceanic Institute with the capability to conduct R&DNBon theNBimpact of feeds and culture system onNBthe product quality of aquacultured shrimp and fish. A taste panel consisting of volunteers from the community and aquaculture industry in Hawaii was trained to evaluate market-size fish and shrimp for various aspects of quality (flavor, texture, color etc.). The taste panel will assist our researchers in their efforts to address the needs of farmers and the aquaculture industry by evaluating the quality of their cultured products to ensure consumer acceptance and confidence. 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). Conquest, L., Laidley, C., Ju, Z. Y. Live feed enrichment for Pacific threadfin, Polydactylus sexfilis: Fatty acid and amino acid profiles. Aquaculture America Annual Meeting, Las Vegas, 13-15 February 2006. Conquest, L., Tacon, A. G. J. Utilization of microbial floc in aquaculture systems: A review. Aquaculture America Annual Meeting, Las Vegas, 13-15 February 2006. Dominy, W.G. 2006. Oceanic Institute: An overview of its goals, departments and achievements. In: Nutritional Biotechnology in the Feed and Food Industries. Proceedings of Alltechs 22nd Annual Symposium, T.P. Lyons, K.A. Jacques, and J.M. Hower, editors. Nottingham University Press, Nottingham, United Kingdom. P. 413-417. Dominy, W.G., Tan, R. K. H., Akiyama, D., Bewley, W. H. 2005. Pelleting process for shrimp feeds. In: Feed Manufacturing Technology V, Chapter 26, Aquatic Feed. Eileen K. Schofield (Technical Editor). American Feed Industry Association,
Arlington, Virginia. P. 288-291. Forster, I., Dominy, W. 2005. Supplemental amino acids in shrimp feeds. International Aquafeed 8(4):8. Ju, Z.Y., Conquest, L., Divakaran, S., Kamarei, R. Amino acid composition of microbial flocs from recirculating shrimp raceways. Aquaculture America Annual Meeting, Las Vegas, 13-15 February 2006. Scientific Publications Izquierdo, M., Forster, I., Divakaran, S., Conquest, L., Decamp, O., Tacon, A. 2006. Effect of green and clear water and lipid source on survival, growth and biochemical composition of Pacific white shrimp Litopenaeus vannamei. Aquaculture Nutrition 12:192-202. Obaldo, L.G., Masuda, R. 2006. Effect of diet size on feeding behavior and growth of Pacific white shrimp, Litopenaeus vannamei. Journal of Applied Aquaculture 18(1):101-110. Verner-Jeffreys, D., Nakamura, I., Shields, R.. 2006. Egg-associated microflora of Pacific threadfin, Polydactylus sexfilis and amberjack, Seriola rivoliana, eggs: Characterisation and
properties. Aquaculture 253(1-4):184-196.
Impacts
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Progress 10/01/04 to 09/30/05
Outputs
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? The United States had an estimated annual trade deficit of $8 billion in edible and non-edible fisheries products in 2002, including $3.4 billion in shrimp imports. This trade deficit is the largest of any agricultural commodity and second only to petroleum for any natural resources product. Expansion of domestic aquaculture production could offset part of this trade deficit and help establish a globally competitive, sustainable (i.e. environmentally non-degrading, technically appropriate, economically viable, and socially acceptable) aquaculture industry in the U.S. Despite considerable scientific advances, optimum feeds, especially those with low or no fish meal, for maximum productivity of Pacific white shrimp (Litopenaeus vannamei) under sustainable culture conditions have not yet been established.
Knowledge of the impact of such feeds and the culture environment on product quality and safety of cultured shrimp is limited. A great need exists to address the above problems in support of the U.S. shrimp industry. In Hawaii every year, disposal of numerous by-products that have potential use in the aquatic feed industry (e.g. meat and bone meal, wheat mill run, coffee bean pulp, macadamia nut presscake, molasses) results in economic loss and potential environmental impacts. Identification and nutrient profiling of potential tropical by-products will permit expansion of aquatic feed research to include the use of these potential ingredients. Pacific threadfin (Polydactylus sexfilis), known locally as moi, and longfin amberjack (Seriola rivoliana), known locally as kahala, are both fast-growing, good-tasting tropical marine carnivorous fish. These fish have been studied at the Oceanic Institute (OI) for the past several years and have shown good market potential. Threadfin and
amberjack culture are growing industries in Hawaii and other tropical regions, and there is a need for research on nutrient requirements and feed development, especially those using fish meal alternatives. The effects of feed type, feeding strategy, and culture environment on flesh quality and growth of these fish require further investigation. Research is needed to ensure that the high consumer preference for these animals is maintained under intensive, sustainable culture conditions. Our proposed project takes an integrated approach in addressing the issue of providing the U.S. with healthful, desirable seafood. This project falls under the Aquaculture National Program 106, in the area of Tropical Aquaculture Research, and has particular relevance to the program component concerning growth, development, and nutrition. The research is also closely allied with other aquaculture program components, such as aquaculture production systems, and quality and safety of aquaculture products
for consumers. The objectives of this project are in alignment with the NP 106 Mission Statement to Conduct high quality, relevant, basic and applied aquaculture research to improve the efficiency, profitability, and sustainability of United States aquaculture, and reduce dependence on imported seafood and threatened ocean fisheries. The potential benefits from attaining these objectives include: an enhanced understanding of vertically-integrated science and technology for production of shrimp, and the model tropical fish, threadfin and amberjack; development of improved feeds with low or no fish meal for tropical species. Improved use of tropical agricultural by-products, feeding technologies, culture management methodology, environmental compatibility, and growth of the U.S. shrimp and tropical marine finfish aquaculture industry; an expanded range of high-quality aquacultured products which will benefit U.S. consumers and food processors, and an increased consumer confidence in the
safety of aquatic foods through quality assurance. Anticipated products of the research include lower-cost, low environmental impact aquaculture feeds with low or no fish meal for use in intensive culture systems, using improved growout feed formulations that employ commercially available feed ingredients and selected by- products of the agriculture industry; reduction in reliance of tropical regions on imported ingredients for aquaculture feed production, improved techniques for assessing advanced physical, chemical, and nutritional quality of feed ingredients and finished feeds, development of feeds and feeding practices for maximum productivity of shrimp and two high-value marine finfish, and improved product quality of cultured shrimp, threadfin, and amberjack. The resulting vertically-integrated science and technology can serve as a contribution to the U.S. aquaculture industry. Primary customers of this research are U.S. aquaculture farmers, aquafeed manufacturers, feed
ingredient suppliers (including those that generate agriculture by-products), and aquatic food processors who are able to directly apply the results of this research. Equipment manufacturers (feed processing; food quality evaluation) and seafood consumers will benefit from improved technology and increased availability of high-quality, affordable aquacultured products. 2. List the milestones (indicators of progress) from your Project Plan. Note: Some modifications to our research became necessary due to insufficient numbers of fingerling threadfin (moi) being available for our planned research tasks. This in turn was due to unexpected mortalities of larval moi in the Institutes Finfish Department hatchery, and we sought permission from ARS to modify some elements of our research to help address this issue from the point of view of aquatic feeds and nutrition. Formal approval was received from ARS for the following changes: Replace Task 4.2, Investigate Taurine Synthesis in
Shrimp and Fish under Sub-objective 4, with four trials on nutrition of Polydactylus sexfilis, also known as Pacific threadfin or moi, at the larval stage. Expand Sub-objective 5, Nutrition for Maximum Productivity and Use of Alternatives to Fish Meal, to include the four trials noted above on larval moi nutrition. These four trials, outlined below in Sub-objective 4, are also reflected in the text of Question 3 of this report. In addition to research modifications in FY 2004-2005, the tasks in our Research Plan for the coming year (October 2005-September 2006) also reflect the enhanced focus on various aspects of moi feeds and nutrition. Most of this years work with moi will take place under Sub-Objective (SO) 5, and consequently, the anticipated milestones associated with this research are listed below under Sub-objective 5. Objective I: Develop Commercially Viable Feeds and Processing Methods Using Tropical Agricultural By-Products Sub-objective 1. Tropical Agricultural
By-Products and Ingredients Characterization. Sub-objective 2. Feed Processing. Sub-objective 3. Feed Development and Characterization. Objective II: Establish Nutritional and Biochemical Strategies for Maximizing Larval and Growout Productivity and Reducing Fish Meal in Feeds for Tropical Species Sub-objective 4. Nutritional Biochemistry and Microbiology of Species and Culture Systems. Sub-objective 5. Nutrition for Maximum Productivity and Use of Alternatives to Fish Meal. Objective III: Establish Product Quality of Species Reared on Developed Feeds Sub-objective 6. Product Quality of Cultured Species Reared on Developed Feeds. 15 Months (January 2005 - April 2006) SO = Sub-Objective SO-1. Tropical Agricultural By-Products and Ingredients Characterization: Completed by-product identification. Completed by-product ingredient characterization. SO-2. Feed Processing: Completed pellet moisture variation, water stability, and pellet durability (PDI) analyses. SO-3. Feed
Development and Characterization: Commercially viable diet formulated for shrimp. Protocol for attractant testing completed. SO-4. Nutritional Biochemistry and Microbiology of Species and Culture Systems: Shrimp cultured in raceways to produce and collect floc. Fractionation of floc in aqueous and solvent phase to obtain fractions (A, B, C, etc.) complete. Pacific Threadfin (Moi) Hatchery Trials 1 through 4: 1. Examine the role and effects of background algae during the rotifer feeding phase of moi larval development and its impact on hatchery survival and early fingerling growout. 2. Examine the effects of rotifer enrichment protocols on moi larval development and hatchery survival. 3. Compare moi production in the OI finfish hatchery with that in the newly opened Pilot Production hatchery facility. 4. Examine the effect of rotifer density on moi production output in the newer Pilot Production facility. SO-5. Nutrition for Maximum Productivity and Use of
Alternatives to Fish Meal: Methionine and lysine requirement studies with threadfin completed. Reduction by half of fish meal from shrimp feeds in zero-water exchange systems. SO-6. Product Quality of Cultured Species Reared on Developed Feeds: Methionine and lysine effect on fish quality evaluated. Dietary lipid and cholesterol effect on shrimp quality determined. 30 Months (April 2006 - July 2007) SO-1. Tropical Agricultural By-Products and Ingredients Characterization: Diet formulation with by-product ingredients completed. SO-2. Feed Processing: By-product processing protocols completed. Lab-scale processing modifications completed. SO-3. Feed Development and Characterization: Physical and chemical analysis of diets completed. Optimal diet for shrimp determined. SO-4. Nutritional Biochemistry and Microbiology of Species and Culture Systems: Fractions A, B, C, etc. tested in shrimp feeds. Chromatographic separation to obtain fractions a, b, c, etc. completed. SO-5. Nutrition
for Maximum Productivity and Use of Alternatives to Fish Meal: Fatty acid requirements for fish evaluated. Develop a formulated broodstock diet for Pacific threadfin (moi). Determine effect of diet and enrichments on nutritional biochemistry of s- type rotifers. Determine biochemical changes related to diet during larval development of moi. Develop weaning and nursery diets for moi. Initial determination of digestive enzyme ontogeny regulation in first feeding larval moi. SO-6. Product Quality of Cultured Species Reared on Developed Feeds: Effect of exercise and feed with required lipid on fish quality completed. Effect of dietary fatty acid on fish quality completed. Effect of best attractants and optimum feeding strategies on fish and fish quality determined. 45 Months (July 2007 - October 2008) SO-1. Tropical Agricultural By-Products and Ingredients Characterization : (Specific milestones not anticipated at this stage for this sub-objective. ) SO-2. Feed Processing: (Specific
milestones not anticipated at this stage for this sub-objective. ) SO-3. Feed Development and Characterization: Shrimp diet recommended to industry. Commercially viable diet formulated for fish. Attractants identified. SO-4. Nutritional Biochemistry and Microbiology of Species and Culture Systems: Fractions a, b, c, etc. tested in shrimp feeds. Structural elucidation of fractions a, b, c, etc. competed. Feeding trials to establish sparing effect of sulfur amino acids completed. SO-5. Nutrition for Maximum Productivity and Use of Alternatives to Fish Meal: Lysine requirement studies completed (amberjack). Protein:energy ratios in diets for fish optimized. Fish meal and oil reduced by half in fish diets. SO-6. Product Quality of Cultured Species Reared on Developed Feeds: Optimum protein:energy ratios effect on fish quality determined. Reduced fish meal / fish oil effect on fish quality completed. 60 Months (October 2008 - January 2010) SO-1. Tropical Agricultural By-Products and
Ingredients Characterization : Optimal by-product inclusion in diet determined. SO-2. Feed Processing: Recommend diet/process to industry. SO-3. Feed Development and Characterization: Optimal diet for fish determined. SO-4. Nutritional Biochemistry and Microbiology of Species and Culture Systems: Pilot-scale evaluation of shrimp growth-promoting compounds completed. SO-5. Nutrition for Maximum Productivity and Use of Alternatives to Fish Meal: Effects of exercise and lipid level on fish established. Fish meal and oil reduced by 2/3 in fish diets. SO-6. Product Quality of Cultured Species Reared on Developed Feeds: Correlation of fish quality with protein/energy ratios and amino acids completed. Correlation of fish quality with dietary lipid, fatty acids, and exercise completed. Quality of shrimp and fish reared on commercially viable feeds established. 3a List the milestones that were scheduled to be addressed in FY 2005. For each milestone, indicate the status: fully met,
substantially met, or not met. If not met, why. 1. Tropical agricultural by-product identification. Milestone Substantially Met 2. Tropical agricultural by-product characterization. Milestone Substantially Met 3. Pellet moisture variation, water stability, and pellet durability (PDI) analyses. Milestone Substantially Met 4. Commercially viable prototype diet formulated for shrimp. Milestone Not Met Other 5. Protocol for attractant testing. Milestone Fully Met 6. Shrimp cultured in raceways to produce and collect floc. Milestone Fully Met 7. Fractionation of floc in aqueous and solvent phase to obtain fractions (A, B, C, etc.). Milestone Fully Met 8. Role and effects of background algae during the rotifer feeding phase of moi larval development, and impact on hatchery survival and early fingerling growout. Milestone Substantially Met 9. Examine the effects of rotifer enrichment protocols on moi larval development and hatchery survival. Milestone Not Met Other 10. Compare moi
production in the Oceanic Institute finfish hatchery with that in the newly opened Pilot Production hatchery facility. Milestone Substantially Met 11. Examine the effect of rotifer density on moi procution output in the new Pilot Production facility. Milestone Not Met Other 12. Methionine and lysine requirement studies with threadfin. Milestone Substantially Met 13. Reduction by half of fish meal from shrimp feed in low-water exchange systems. Milestone Substantially Met 14. Methionine and lysine effect on fish quality. Milestone Substantially Met 15. Dietary lipid and cholesterol effect on shrimp quality. Milestone Fully Met 3b List the milestones that you expect to address over the next 3 years (FY 2006, 2007, and 2008). What do you expect to accomplish, year by year, over the next 3 years under each milestone? 30 Months (April 2006 - July 2007) SO-1. Tropical Agricultural By-Products and Ingredients Characterization : Diet formulation with by-product ingredients completed. SO-2.
Feed Processing: By-product processing protocols completed. Lab-scale processing modifications completed. SO-3. Feed Development and Characterization: Physical and chemical analysis of diets completed. Optimal diet for shrimp determined. SO-4. Nutritional Biochemistry and Microbiology of Species and Culture Systems: Fractions A, B, C, etc. tested in shrimp feeds. Chromatographic separation to obtain fractions a, b, c, etc. completed. SO-5. Nutrition for Maximum Productivity and Use of Alternatives to Fish Meal: Fatty acid requirements for fish evaluated. Develop a formulated broodstock diet for Pacific threadfin (moi). Determine effect of diet and enrichments on nutritional biochemistry of s- type rotifers. Determine biochemical changes related to diet during larval development of moi. Develop weaning and nursery diets for moi. Initial determination of digestive enzyme ontogeny regulation in first feeding larval moi. SO-6. Product Quality of Cultured Species Reared on Developed
Feeds: Effect of exercise and feed with required lipid on fish quality completed. Effect of dietary fatty acid on fish quality completed. Effect of best attractants and optimum feeding strategies on fish and fish quality determined. 45 Months (July 2007 - October 2008) SO-1. Tropical Agricultural By-Products and Ingredients Characterization : (Specific milestones not anticipated at this stage for this sub-objective. ) SO-2. Feed Processing: (Specific milestones not anticipated at this stage for this sub-objective. ) SO-3. Feed Development and Characterization: Shrimp diet recommended to industry. Commercially viable diet formulated for fish. Attractants identified. SO-4. Nutritional Biochemistry and Microbiology of Species and Culture Systems: Fractions a, b, c, etc. tested in shrimp feeds. Structural elucidation of fractions a, b, c, etc. competed. Feeding trials to establish sparing effect of sulfur amino acids completed. SO-5. Nutrition for Maximum Productivity and Use of
Alternatives to Fish Meal: Lysine requirement studies completed (amberjack). Protein:energy ratios in diets for fish optimized. Fish meal and oil reduced by half in fish diets. SO-6. Product Quality of Cultured Species Reared on Developed Feeds: Optimum protein:energy ratios effect on fish quality determined. Reduced fish meal / fish oil effect on fish quality completed. 60 Months (October 2008 - January 2010) SO-1. Tropical Agricultural By-Products and Ingredients Characterization : Optimal by-product inclusion in diet determined. SO-2. Feed Processing: Recommend diet/process to industry. SO-3. Feed Development and Characterization: Optimal diet for fish determined. SO-4. Nutritional Biochemistry and Microbiology of Species and Culture Systems: Pilot-scale evaluation of shrimp growth-promoting compounds completed. SO-5. Nutrition for Maximum Productivity and Use of Alternatives to Fish Meal: Effects of exercise and lipid level on fish established. Fish meal and oil reduced by
2/3 in fish diets. SO-6. Product Quality of Cultured Species Reared on Developed Feeds: Correlation of fish quality with protein/energy ratios and amino acids completed. Correlation of fish quality with dietary lipid, fatty acids, and exercise completed. Quality of shrimp and fish reared on commercially viable feeds established. 4a What was the single most significant accomplishment this past year? Effectiveness of an inexpensive form of methionine for use in shrimp diets: Different forms of methionine (including L-methionine, DL-methionine and HMTBA [(2-hydroxy-4-methylthio) butanoic acid]) were shown to be equal in their ability to meet the needs for dietary methionine of shrimp grown in low-water exchange pond-like conditions. A trial was conducted in order to find an inexpensive form of methionine (HMTBA) that could be used to improve the nutritional quality of diets containing high levels of soybean meal, which is an inexpensive source of protein but contains a low level of
methionine. In this trial, the growth rate of shrimp fed a control diet containing low methionine (high soybean meal) was compared with the same diet supplemented with a fixed level of each of three forms of methionine. Use of HMTBA could enable formulation of commercial shrimp feeds with higher levels of soybean meal in replacement of fish meal, while meeting the nutritional needs of the animal. 4b List other significant accomplishments, if any. Identification of bioactive growth-promoting compounds in shrimp culture systems: In ongoing research toNBisolate bioactive compounds in the microbial community (floc) of shrimp culture systemsNBthat are beneficial to shrimp nutrition, wet floc was collected from shrimp raceways, freeze-dried, fractionated with aqueous and organic solvents, and the components analyzed, indicating 12% dry matter with 32% crude protein and 2% crude fat. The research was undertaken in an effort to find new ways of augmenting shrimp growth by isolation,
identification, and quantification of growth-promoting bioactive compounds in the floc from shrimp culture systems, and to use the methods established in the course of this work for future shrimp growout trials. NB In the work to date, new methods have been developed to fractionate the floc and isolate various components via HPLC and GC analysis, including carotenoid pigments, bioactive sterols, peptides and phenolic antioxidants, amino acids, fatty acids, and vitamins. This research has the potential to assist feed manufacturers in developing improved feeds for better shrimp performance. Effect of dietary lipid and cholesterol on quality of cultured shrimp: An experiment in which shrimp were fed a diet containing a veryNBlow level of cholesterol and a highNBlevel of docosahexaenoic acid (DHA) resulted in a lower level of cholesterol and a higher level of DHA in theNBtail muscle tissue of market-size shrimp, compared to shrimp fed a commercial control diet.NB Research was needed to
determine whether the nutritional quality of cultured shrimp could be improved through modifications in the feed formulation, as a means of providing heart-healthy seafood to consumers who are aware of the importance of diet in overall health.NB The trial was conducted withNB9 g shrimpNB(initial weight),NBfed with experimental and control dietsNBand grown inNBoutdoor culture tanks (1100 liters functional volume, 75 animals/tank, 2 L/min flow rate) to the market sizes of 20 g and 27 g, followed by evaluation of the effects of diet on cholesterol and DHA content of shrimp tail muscle tissue.NB Results clearly showed that shrimp product quality can be improved (i.e., positive quality attributes enhanced, and negative attributes reduced) by substantially manipulating the ingredients incorporated into the diet. 5. Describe the major accomplishments over the life of the project, including their predicted or actual impact. Two academic prototype shrimp feeds (one for indoor culture, one for
outdoor) were developed which provide excellent performance, often outperforming commercial feeds. The commercially viable versions of these feeds are being formulated and tested, after which they will be moved to pilot scale for commercial applications. The problems addressed by this accomplishment include the traditional use in academic feed formulations of ingredients that are costly and not available year-round or in large quantities, or contain too many ingredients in the formulation, which increases the time required for manufacturing, thereby increasing the inventory, need for storage space, number of purchases, and number of tracking and accounting entries and payments. The objective is to simplify the formulation and use common feed ingredients currently available to the feed industry at the minimum cost. Different forms of methionine (including L-methionine, DL-methionine and HMTBA [(2-hydroxy-4-methylthio) butanoic acid]) were shown to be equal in their ability to meet
the needs for dietary methionine of shrimp grown in low-water exchange pond-like conditions. Use of HMTBA could enable formulation of commercial shrimp feeds with higher levels of soybean meal in replacement of fish meal, while meeting the nutritional needs of the animal. In ongoing research toNBisolate bioactive compounds in the microbial community (floc) of shrimp culture systemsNBthat are beneficial to shrimp nutrition, wet floc was collected from shrimp raceways, freeze-dried, and fractionated into aqueous and solvent phases. Analysis of the components indicated a composition of 12% dry matter, 32% crude protein, and 2% crude fat. In the course of this research, new methods were developed to fractionate the floc and isolate various components via HPLC and GC analysis, including carotenoid pigments, bioactive sterols, peptides and phenolic antioxidants, amino acids, fatty acids, and vitamins. This research has the potential to assist feed manufacturers in producing improved feeds
for better shrimp performance. An experiment in which shrimp were fed a diet containing a veryNBlow level of cholesterol and a highNBlevel of docosahexaenoic acid (DHA) resulted in a lower level of cholesterol and a higher level of DHA in theNBtail muscle tissue of market size shrimp, compared to shrimp fed a commercial control diet.NB Results clearly showed that shrimp product quality can be improved (i.e., positive quality attributes enhanced, and negative attributes reduced) by substantially manipulating the ingredients incorporated into the diet. Note: In the accomplishments listed above, the potential beneficiaries include U.S. aquaculture farmers, aquafeed manufacturers, feed ingredient suppliers (including those that generate agriculture by-products), and aquatic food processors who are able to directly apply the results of this research. Equipment manufacturers (feed processing; food quality evaluation) and seafood consumers, will also benefit from improved technology and
increased availability of high-quality, affordable aquacultured products. 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? A presentation on methionine in feeds for Pacific white shrimp, Litopenaeus vannamei, was given at the annual meeting of the World Aquaculture Society in Bali, Indonesia, in May 2005. Project staff requested and received approval to have aquacultured shrimp, Pacific threadfin, and amberjack included in a revised color poster entitled Hawaii & Pacific Islands Seafood, produced by the State of Hawaii, Department of Business, Economic Development and Tourism, Ocean Resources Branch. The poster presents photographs of all the major seafood species in the islands, detailed species information, cooking instructions, yield % of round
weight, buying and cooking tips, and nutritional information. The project was given 5,000 copies, on which Oceanic Institute stickers can be placed prior to distribution to interested individuals and organizations in the aquaculture and seafood industries. The Oceanic Institutes Nutrition Department (which is responsible for this project) will be represented by two staff members when they attend the American Feed Industry Association conference, expo and Aquaculture Committee meeting, to be held in Kansas City, Missouri in September of 2005. One of them will also help staff the AFIA Aquaculture Committee booth at the expo that will explain to booth visitors about the OI Nutrition Departments research and activities, and how it supports and relates to the feed industry. Both AFIA and its Aquaculture Committee members are important recipients of the Nutrition Departments research and activities. 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). Divakaran, S., Forster, I., Velasco, M. 2004. Limitations on the use of shrimp Litopenaeus vannamei midgut gland extract for the measurement of in vitro protein digestibility. Aquaculture 239:323-329. Divakaran, S., Moss, S.M. 2004 In vitro evidence of laminarinase activity in the digestive gland of juvenile Pacific white shrimp Litopenaeus vannamei. Journal of the World Aquaculture Society 35(4):546- 550. Kamarei, A.R., Trygstad, C. 2004. Designing clinical trials to substantiate claims. Food Technology 58(10):28-35.
Impacts
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Publications
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