Source: UNIVERSITY OF NORTH CAROLINA - WILMINGTON submitted to NRP
CONTROLLED BREEDING, LARVICULTURE, AND INTENSIVE GROWOUT OF HIGH VALUE MARINE FINFISH SPECIES FOR U.S. AGRICULTURE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
OTHER GRANTS
Reporting Frequency
Annual
Accession No.
0213722
Grant No.
2008-38854-18896
Cumulative Award Amt.
(N/A)
Proposal No.
2008-03127
Multistate No.
(N/A)
Project Start Date
Aug 1, 2008
Project End Date
Jul 31, 2012
Grant Year
2008
Program Code
[NQ]- Mariculture, NC
Recipient Organization
UNIVERSITY OF NORTH CAROLINA - WILMINGTON
601 SOUTH COLLEGE ROAD
WILMINGTON,NC 28403
Performing Department
(N/A)
Non Technical Summary
Southern flounder (SF, Paralichthys lethostigma) Based on research conducted in NC, culture technologies for SF are being transferred to commercial users. Solutions to the following constraints are needed to expand commercial production: Currently, we rely on wild-caught broodstock, but domesticating SF will require breeding cultured progeny for improved performance. F1 females are difficult to spawn using hormone therapies developed for wild-caught adults, and alternative hormone therapies for F1 females are needed to facilitate selective breeding programs and establish improved lines of SF. Economic analysis of a SF recirculating aquaculture system growout operation showed that fingerling costs represent 24-29% of production costs, with live rotifers representing the largest component (~15%) of fingerling production costs. Developing artifical microdiets in substitution of rotifers can significantly lower fingerling costs to farmers. The proposed research aims to enhance the development of the SF aquaculture industry in NC by solving key constraints to broodstock husbandry and to cost-effective production of fingerlings. Specific objectives are as follows: 1. Standardize hormone therapies for induced breeding of F1 females. 2. Reduce fingerling production costs by minimizing reliance on live prey through developing artificial microdiets for co-feeding and early weaning. Black Sea Bass (BSB, Centropristis striata) BSB is now in the transitional phase between research and commercial production in the eastern US. The current proposed work addresses a critical constraint to the development of a viable marine finfish culture industry in the U.S.: The use of wild-caught fish for production of fish meal as a source of protein and oil for aquaculture diets threatens wild stocks and creates economic uncertainty in aquaculture ventures. The goals of the proposed research are to promote sustainable marine finfish aquaculture by developing lower-cost feeds that rely less on dietary inclusion of fish meal. For BSB, specific objectives are as follows: 3. Evaluate substitution limits of alternative proteins (underutilized plant and animal by-products) as a fish meal replacement in diets. 4. Determine the effects of these feeds on fish biochemical composition. Red porgy We have made significant progress in understanding the spawning and larval culture requirements of RP, and post-metamorphic juveniles have been produced in our laboratory. We propose to optimize hatchery technologies and lower fingerling production costs for large-scale production of RP fingerling for pilot commercial nursery and growout projects. Specific objectives are as follows: 5. Optimize enrichment methods for live feeds. 5.1 Compare practical enrichment media, including concentrated non-viable microalgae and state-of-the-art commercial formulations. 5.2 Determine the effects of feeding cold-banked prey on larval performance. 6. Transfer technologies to the commercial end-users.
Animal Health Component
75%
Research Effort Categories
Basic
10%
Applied
75%
Developmental
15%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3013719108020%
3023719101020%
3023719105020%
7013719101020%
7013719105020%
Knowledge Area
302 - Nutrient Utilization in Animals; 301 - Reproductive Performance of Animals; 701 - Nutrient Composition of Food;

Subject Of Investigation
3719 - Other cultured finfish;

Field Of Science
1010 - Nutrition and metabolism; 1080 - Genetics; 1050 - Developmental biology;
Goals / Objectives
Research focuses on three candidate species for aquaculture: southern flounder (SF, Paralichthys lethostigma), black sea bass (BSB, Centropristis striata), and red porgy (RP, Pagrus pagrus). We propose to address critical contraints to commercial cultivation of each species. Southern Flounder The proposed research aims to support and enhance the development of the nascent SF aquaculture industry in NC by solving key biotechnical constraints to broodstock husbandry and to cost-effective, large-scale production of fingerlings for fish farming. For SF, specific objectives (1,2) are as follows: 1. Standardize hormone therapies for induced breeding of first-generation (F1) females. 2. Reduce fingerling production costs by minimizing reliance on live prey through developing artificial microdiets for co-feeding and early weaning. Black Sea Bass The goals are to promote sustainable marine finfish aquaculture by developing lower-cost feeds that rely less on dietary inclusion of fish meal. For BSB, specific objectives (3, 4) are as follows: 3. Evaluate substitution limits of alternative proteins (underutilized plant and animal by-products) as a fish meal replacement in BSB diets under controlled laboratory conditions. 3.1. Replace menhaden fish meal by solvent extracted soybean meal supplemented with coated crystalline amino acids in BSB diets. 3.2. Replace menhaden fish meal by poultry by-products and fermented poultry by-products in BSB diets. 3.3. Replace menhaden fish meal by meat and bone meal in BSB diets. 3.4. Formulate different cost effective diets using a combination of different alternative protein sources (e.g. soybean meal, poultry by-product, and meat and bone meal) and determine their effects on growth and body composition of BSB. 4. Determine the effects of these feeds on fish biochemical composition. Red porgy We will optimize hatchery technologies and lower fingerling production costs for large-scale production of RP fingerling for pilot commercial nursery and growout projects. Specific objectives (5) are as follows: 5. Optimize enrichment methods for live feeds. 5.1 Compare practical enrichment media, including concentrated non-viable microalgae (N. oculata) and state-of-the-art commercial formulations. 5.2 Determine the effects of feeding cold-banked prey on larval performance. 6. Transfer technologies to the commercial end-users. Anticipated benefits Methods for reliable spawning of F1 SF females will help establish selective breeding programs for domestication of SF as an aquacultured species. A high quality microdiet will minimize reliance on live prey to generate reliable and cost-effective production output. Information on the substitution limits of a variety of alternative protein sources for menhaden fish meal will help develop environment-friendly, cost-effective diets for BSB that produces high quality flesh (especially omega-3 fatty acids) for markets. Information on the biochemical composition of fish grown on different alternative protein sources will establish quantitative factors that affect taste and quality. Optimizing prey enrichment protocols will generate reliable and cost-effective production output for RP.
Project Methods
Southern flounder (SF) 1. Standardize hormone therapies for induced breeding of F1 females. We will compare mammalian and piscine neuropeptide analogs in different dose rates and modes of hormone administration on ovulatory response, egg production, and egg quality in F1 females. 2. Reduce fingerling production costs by minimizing reliance on live prey through developing artificial microdiets for co-feeding and early weaning. Growth performance, survival and stress resistance of larvae will be evaluated by feeding formulated zein microbound diets (MBD) containing different protein sources (e.g. menhaden meal, herring meal, squid meal, krill meal, soybean peptide, and casein) and high quality lipid, carbohydrate, vitamins, minerals. Growth performance will also be compared on MBD containing attractants or supplemented with crystalline amino acids to improve appetency and/or amino acid profile. Black Sea Bass (BSB) 3. Evaluate substitution limits of alternative proteins (underutilized plant and animal by-products) as a fish meal replacement in diets. We will develop cost-effective practical diet formulations to support optimal growth of juvenile and subadult BSB. Through laboratory studies, we will determine the effects of amino acid (methionine and lysine) supplementation in soybean based diets. The substitution limits of alternative proteins for menhaden fish meal in diets for BSB will be delineated. We will also formulate and test diets that combine animal and plant ingredients for optimal protein and lipid profiles. 4. Determine the effects of these feeds on fish biochemical composition. Diets will be analyzed for protein, fat, fiber, ash, calories, fatty acid and amino acid profiles. Fish will be analyzed for proximate composition, fatty acids, cholesterol, and amino acids to establish quantitative factors that affect taste and fish quality. Red porgy 5. Optimize enrichment methods for live feeds. 5.1 Compare practical enrichment media, including concentrated non-viable microalgae and state-of-the-art commercial formulations. Performance (growth, survival, stress resistance) of larvae fed rotifers enriched with different practical media will be compared through d15ph. 5.2 Determine the effects of feeding cold-banked prey on larval performance. 6. Transfer technologies to the commercial end-users. Anticipated benefits Methods for reliable spawning of F1 SF females will help establish selective breeding programs for domestication of SF as an aquacultured species. A high quality microdiet will minimize reliance on live prey to generate reliable and cost-effective production output. Information on the substitution limits of a variety of alternative protein sources for menhaden fish meal will help develop environment-friendly, cost-effective diets for BSB that produces high quality flesh (especially omega-3 fatty acids) for markets. Information on the biochemical composition of fish grown on different alternative protein sources will establish quantitative factors that affect taste and quality. Optimizing prey enrichment protocols will generate reliable and cost-effective production output for RP.

Progress 08/01/08 to 07/31/12

Outputs
Target Audience: Target audiences includeresearchers, commercial farmers, educators and public policy makers to whom project results were disseminated through conference presentations, press releases and publications. Target audiences also include the graduatestudents listed in the progress reports who participated in the experimental work and derived MS thesis projects; undergraduate students who conducted aquaculture practicums and received experiential learning opportunities; commercialsouthern flounder and black sea bass producers and feed manufacturerswho benefited from the culture technologies developed during this project;andprimary, secondary and college students from various institutionsand the general public who toured the UNCW Aquaculture Facility during the period of this project. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? The following students conducted thesis research and received Masters of Marine Science degrees or received training under support by this USDA-NIFA project: Graduate student: Katherine B. Sullivan (MS), conducted experiments on replacement of fish meal in black sea bass diets with soybean meal, poultry meal and meat and bone meal. Results were incorporated into a MS Thesis in Marine Science. MS degreereceived Fall 2008. Graduate student: Matthew R. Dawson (BS): conducted experiments on replacement of fish meal in black sea bass diets with poultry meal and combined alternative protein sources. Results were incorporated into a MS Thesis in Marine Science. MS degreereceived in Fall 2012. Graduate student: Walker Wright-Moore (BS), assisted with induced spawning experiments for southern flounder as well as with nutrition experiments with black sea bass. Results will be incorporated into a MS Thesis in Marine Science. Anticipated graduation Fall 2013. Graduate student: Andrew D. Ostrowski, assisted with red porgy larviculture and live prey enrichment studies. Undergraduate students: T. Gibson, R. Doyle, A. Markwith, C.Kelly, assisted with experiments. How have the results been disseminated to communities of interest? Results have been presented to local, regional, national and international aquaculture conferences and published in peer-reviewed scientific journals for dissemination to the aquaculture community, including researchers, commercial farmers, educators and public policy makers. Results have also been disseminated to the general public through articles in various media: Article entitled “Ocean fish, fresh from the farm” by Wade Rawlins, Raleigh News and Observer, Jan. 18, 2008. Article entitled “Ocean fish, served fresh from the farm”, in redOrbit, 22 Jan. 2008. Article entitled “From fish farm to plate: aquaculture center on the brink of commercialization, by Jennifer Roush, Lumina News, Jan. 24, 2008 Article entitled “Fish farm lab at Wrightsville shows promise: UNCW-N.C. State project could boost fish stocks, and economy, Published: Saturday, January 19, 2008, Wilmington Star-News. Article entitled “U.S. lags in marine agriculture; can SouthCoast captitalize on this market?, by The Cape Cod Times, Jan. 26, 2008. Article entitled “Beach Bites: Wandering Eye, Flounder Research and Development” by Jessica Haywood, in Wrightsville Beach Magazine, August 2008. Article entitled “Lawmakers hear about aquaculture: UNCW officials, fish farmers talk to new committee in Raleigh, by Joe Johnson, Wilmington Star-News, April 17, 2009. TV feature entitled “NC Rising: Biotechnology”, by Elizabeth Wilder, on UNC-TV: NC Now, May 11, 2009. Article entitled “MARBIONC spawns entrepreneurs” by Jennifer Woodward, Impact Magazine, May 2009. Article entitled “Blue Thumbs: NC's Aquaculture Industry”, by Jennifer Woodward, Impact Magazine, May 2009. Article entitled “Open house offers look at Sturgeon City’s Riverworks” by Lindell Kay, Jacksonville Daily News, May 7, 2009. Article entitled “Soybean protein used for partial replacement of fish meal in the diet of juvenile black sea bass” in Hatchery International, p. 27, May/June 2009. Article entitled “Researchers use effluent from a marine recirculating aquaculture system to grow phytoplankton, in Hatchery International, p. 40, May/June 2009. Article entitled “Farming saltwater fish goal of UNCW project”, by James Gallagher, In Triangle Business Journal (Raleigh, NC). Coastal Business Sections. February 19, 2010. Article entitled “Aquaculture pioneer grows fish industry”, by Allison Lee Satake, in Greater Wilmington Business Journal, October 15-28, 2010. Results have also been disseminated to commercial southern flounder and black sea bass farmers who are using the culture technologies developed during this project. This includes embryos, fingerlings and feeds, based on induced spawning, hatchery and alternative protein sources technologies. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Specific projects objectives and accomplishment are as follows: 1. Standardize hormone therapies for induced breeding of F1southern flounder. There were significant effects of hormone dose on spawning performance (i.e.,production of viable larvae), with maximum production observed at the low dose (5 µg/kg bw) for both LHRHa and sGnRHa. Larval production was higher with LHRHa in F1 females and with sGnRHa in wild females. 2. Reduce southern flounder fingerling production costs by minimizing reliance on live prey through developing artificial microdiets. A UNCW-formulatedmicrobound diet containing menhaden, squid, krill, herring meal and attractants, co-fed with one half live feed rations from d11ph, produced no difference in survival compared to a feeding regimen usinglive feed alone and could therefore be used to reduce live feed costs during larval rearing. 3. Evaluate substitution limits of underutilized plant and animal by-products as a fish meal replacement in diets. 3.1. Replacement of menhaden fish meal by solvent extracted soybean meal supplemented with amino acids. The optimum level of fish meal protein replacement with solvent extracted soybean protein was 70% with 7.5% squid meal and 5% krill meal and with or without supplemental methionine and lysine in the diet. 3.2. Replacement of menhaden fish meal by poultry by-products and fermented poultry by-products. Menhaden fish meal protein could be replaced up to 70% by soybean protein in juvenile diets when supplemented with small amount of squid meal, krill meal, fermented poultry by products or methionine and lysine without impairing growth. Poultry by-product meal is a promising alternative protein substitute for fish meal protein for sustainable diets. 3.3. Replacement of menhaden fish meal by meat and bone meal. Fish meal protein may be replaced by soybean meal protein (without supplemental Met and Lys) at levels up to 70% in the diet. Fish meal replacement by meat-and-bone meal and poultry-by-product meal at levels of at least 30% and 60%, respectively, were successful. 3.4. Formulation of different cost effective diets using a combination of different alternative protein sources (e.g. soybean meal, poultry by-product, and meat and bone meal) and their effects on growth and body composition. The optimal diet producing no significant reduction in performance contained only 6% fish meal (70% fish meal protein replacement) in combination with soybean meal, poultry by-product meal, meat and bone meal, squid meal, and krill meal. A practical low fish meal diet containing a mixture of alternative protein sources is possible for juvenile black sea bass. 4. Determine the effects of thesefeeds on fish biochemical composition. Black sea bass fed the 70% meat and bone meal protein replacement diet and diets containing poultry byproduct mealand meat and bone mealprotein produced lower EPA, DHA, and total n-3 PUFA concentrations in whole body tissues than fish fed the control fish meal diet. 5. Optimize enrichment methods for live feeds for larval red porgy. Performance was better in the Algamac treatment and the DC DHA Selco treatment than in the control. Survival was significantly higher in the DHA Selco treatment than in Algamac 3000 and the control. Prey enrichment protocols have been established for red porgy that will generate more cost-effective juvenile production output. Overall results have identified improved prey enrichment protocols and developed a formulated microdietusing state of the art products and techniques to enable simpler rearing protocols for red porgy and more reliable production output. These results methodically advance larval culture technologies for red porgy developed at UNCW.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2008 Citation: Sullivan, K.B. M.S. Alam, and W.O. Watanabe. 2008. Replacement of fish meal by soybean meal, poultry byproduct meal and meat and bone meal in diets for juvenile black sea bass Centropristis striata. Book of Abstracts, Aquaculture America 2008, Orlando, Florida
  • Type: Theses/Dissertations Status: Published Year Published: 2008 Citation: Sullivan, K.B. 2008. Replacement of fish meal by alternative protein sources in diets for juvenile black sea bass. Masters Thesis, University of North Carolina Wilmington, Center for Marine Science.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2008 Citation: Alam, M.S., Watanabe W.O., Sullivan, K.B. 2008. Recent progress on development of nutritionally-balanced, cost-effective and environmentally-friendly diets for the culture of black sea bass Centropristis striata and southern flounder Paralichthys lethostigma. Book of Abstracts, Eighth Joint Meeting of the Seafood Science and Technology Society and Atlantic Fisheries Technology Conference in conjunction with the 1st NC Marine Biotechnology Symposium 2008. Wrightsville Beach, NC, Oct 20-23, 2008.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2009 Citation: Alam, M.S., W.O. Watanabe, K.B. Sullivan and W. D. Wright-Moore. 2009. Partial and full replacement of menhaden fish meal protein by soybean meal protein supplemented with or without amino acids and attractants in the diets of juvenile black sea bass Centropristis striata. Book of Abstracts, Aquaculture America 2009, Seattle, WA, USA
  • Type: Conference Papers and Presentations Status: Published Year Published: 2011 Citation: Alam, M.S., Watanabe, W.O., Daniels, H.V., Ferket, P., Gill, A., Rezek, T.C, Myers A.R, Dawson, M. 2011. Replacement of fish meal protein by soybean meal protein supplemented with or without fermented poultry by products, squid meal, krill meal and amino acids in the diets of juvenile black sea bass Centropristis striata. Aquaculture America 2011. Book of Abstracts. Aquaculture America. New Orleans, Louisiana, Feb 28-Mar 3, 2011.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2011 Citation: Dawson, M.R., M.S. Alam, and W.O. Watanabe. 2011. Effects of poultry by-product meal protein substitution for fish meal protein on growth performance, body composition, and digestibility of juvenile black sea bass Centropristis striata. Book of Abstracts. Aquaculture 2011, New Orleans, Louisiana, March 02-04, 2011.
  • Type: Journal Articles Status: Published Year Published: 2011 Citation: Alam, M.S., Watanabe, W.O., Myers, A.R., Rezek, T.C., Carroll, P.M., Longfellow, S. 2011. Effects of replacement of menhaden fish meal protein by solvent extracted soybean meal protein supplemented with or without L-methionine and L-lysine on growth performance and body composition of juvenile southern flounder. North American Journal of Aquaculture 73:350359.
  • Type: Journal Articles Status: Published Year Published: 2012 Citation: Alam, M.S., Watanabe, W.O., Sullivan, K.B., Rezek, T.C., Seaton, P.J. 2012. Replacement of menhaden fish meal protein by solvent extracted soybean meal protein in the diet of juvenile black sea bass Centropristis striata supplemented with or without squid meal, krill meal, methionine and lysine. North American Journal of Aquaculture 74:251-265.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2011 Citation: Watanabe, W.O. 2011. Marine Finfish Aquaculture at UNCW: Research, Commercial Pilot Initiatives and Regulations. Invited presentation to the North Carolina Aquaculture Development Conference, February 11-12 2011, Sheraton Atlantic Beach, NC.
  • Type: Journal Articles Status: Published Year Published: 2011 Citation: Carrier, J.K. III, W.O. Watanabe, M. Harel, T. F. Rezek, P. J. Seaton and T.H. Shafer. 2011. Effects of dietary arachidonic acid on larval performance, fatty acid profiles, stress resistance, and expression of Na+/K+ ATPase mRNA in black sea bass Centropristis striata. Aquaculture 319: 111-121.
  • Type: Conference Papers and Presentations Status: Other Year Published: 2011 Citation: Watanabe, W.O. 2011. UNCW Aquaculture Program: Research, Education and Commercial Pilot Initiatives. Presentation to the North Carolina Farm Bureau Apr. 20, 2011. UNCW Center for Marine Science, Myrtle Grove, NC.
  • Type: Journal Articles Status: Under Review Year Published: 2013 Citation: Alam, M.S., W.O. Watanabe, T.C. Rezek, A.R. Myers, P.M. Carroll and H.V. Daniels. In Review. Growth performance and survival of larval southern flounder Paralichthys lethostigma fed commercial microdiets or formulated zein bound microdiets containing different protein sources. Submitted to Aquaculture Research.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Alam, M.S., Watanabe, W.O., Carroll, P.M., Rezek, T.C., Myers A.R., Dawson, M. 2012. Diet Development of Marine Fish at University of North Carolina Wilmington (UNCW). Book of Abstracts, North Carolina Aquaculture Development Conference, Hilton New Burn, New Bern, NC, Feb 09-11, 2012.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2012 Citation: Wright-Moore, W.D., W.O. Watanabe and T.C. Rezek, 2012. Induced spawning of F1 and wild-caught southern flounder with GnRH analogs: effects of hormone type (mammalian vs piscine) and dose on egg production and quality. Book of Abstracts, Aquaculture America 2012, Feb 29-Mar 12, Paris Hotel and Casino, Las Vegas, NV
  • Type: Theses/Dissertations Status: Published Year Published: 2012 Citation: Dawson, Matthew. 2012. Evaluation of alternative protein sources to fish meal in practical diets for juvenile black sea bass Centropristis striata and southern flounder Paralichthys lethostigma. Masters Thesis, University of North Carolina Wilmington, Center for Marine Science.


Progress 08/01/09 to 07/31/10

Outputs
OUTPUTS: 1. Standardize hormone therapies for induced breeding of F1southern flounder females. We compared mammalian and piscine neuropeptide analogs (LHRHa and sGnRHa), dose rates (0, 5, 25, and 50 ug/kg bw) and modes of administration (slow and fast-release pellet implants) on egg production and egg quality in F1 and wild-caught females. 2. Reduce southern flounder fingerling production costs by minimizing reliance on live prey through developing artificial microdiets for larval weaning. Growth performance of larvae was evaluated by feeding formulated zein microbound diets (MBDs) containing different protein sources. In Expt. 1, four MBDs were prepared with a mixture of protein sources (menhaden, squid, krill and herring meal) with or without attractants and were compared with a premium, commercially available MBD. In Expt. 2, the effects of artemia and rotifer exclusion during weaning in larvae fed MBDs were evaluated. 3. Evaluate substitution limits of alternative plant and animal by-products as a fish meal replacement in black sea bass diets under laboratory conditions. 3.1. Replacement of menhaden fish meal by solvent extracted soybean meal supplemented with amino acids. A series of experiments were conducted to test the effect of varying dietary levels of solvent-extracted soybean meal supplemented with or without amino acids and attractants in the diets of juveniles. 3.2. Replacement of menhaden fish meal by poultry by-products and fermented poultry by-products. Experiments were conducted to test the effect of replacing fish meal protein with poultry by-product meal protein and solvent-extracted soybean meal supplemented with or without fermented poultry by-products, squid meal, krill meal and amino acids in the diet of juveniles. 3.3. Replacement of menhaden fish meal by meat and bone meal. An experiment was conducted to test diets that replaced fish meal protein by soybean meal protein at levels from 0-100% and a diet with 60% of the fish meal replaced by poultry-by-product meal and a diet with 30% of the fish meal replaced by meat-and-bone meal. 3.4. Formulation of different cost effective diets using a combination of different alternative protein sources (e.g. soybean meal, poultry by-product, and meat and bone meal) and their effects on growth and body composition. An experiment was conducted to test replacement of fish meal protein by poultry by-product meal protein and by meat and bone meal protein in diets containing a high level of solvent extracted soybean meal protein with and without squid and krill meals. 4. Determine the effects of these feeds on fish biochemical composition. Experimental diets and fish are being analyzed for proximate, fatty acid and amino acid profiles. 5. Optimize enrichment methods for live feeds for larval red porgy. 5.1 Compare practical enrichment media, including concentrated non-viable microalgae and commercial formulations. In Expt. 1, we compared performance of larvae fed rotifers and artemia enriched with different practical media. In Expt. 2, we determined the effects of feeding cold-banked prey on larval performance. PARTICIPANTS: Individuals: Project Director: Dr. Wade O. Watanabe, project coordinator and technical and scientific supervisor, including project administration. Co-Principal Investigator: Dr. Md Shah Alam: technical and scientific supervision for nutrition experiments on alternative protein sources to fish meal in black sea bass diets. Research Assoc.: Patrick M. Carroll; operational assistance on experimental work through facilities design, setup, and maintenance. Research Assoc.: Troy C. Rezek (MS), technical assistance for induced spawning experiments of southern flounder using LHRHa and on larviculture experiments with red porgy. Research Assoc.: Amanda R. Myers (MS), technical assistance with diet studies with southern flounder and black sea bass. Partner Organizations: NC Sea Grant, provided funding support which help leverage induced spawning experiments with southern flounder NOAA Marine Aquaculture Program, provided funding support which helped leverage nutrition experiments with black sea bass Golden Leaf Foundation provided funding support which helped leverage induced spawning experiments with southern flounder. Collaborators and contacts: Dr. James Morris, Center for Coastal Fisheries and Habitat Research (CCFHR), National Ocean Service (NOS), NOAA, provided red porgy embryos which were used for larval rearing experiments on live prey enrichment. Dr. Harry V. Daniels (North Carolina State University), received southern flounder embryos that were produced as a product of induced spawning experiments. These embryos were used for experimental purposes. Mr. Keith Hairr (Carolina Flounder LLC), received southern flounder embryos produced in induced spawning experiments. These embryos were used for commercial hatchery purposes at CF. Mr. Shawn Longfellow (Blue Ocean Farms LLC), received southern flounder fingerlings from CF that were produced in induced spawning experiments. Fingerlings were used for commercial growout and marketing at BOF. Mr. Ted Davis (Aqua Plantations LLC), received black sea bass fingerlings from UNCW that were produced in nutrition experiments. Fingerlings were used for commercial growout and marketing purposes at AP. Training or professional development: Graduate student: Katherine B. Sullivan (MS), conducted experiments on replacement of fish meal in black sea bass diets with soybean meal, poultry meal and meat and bone meal. Results were incorporated into a MS Thesis in Marine Science. MS degree was received Fall 2008. Graduate student: Matthew R. Dawson (BS): conducted experiments on replacement of fish meal in black sea bass diets with poultry meal and combined alternative protein sources. Results will be incorporated into a MS Thesis in Marine Science. Graduate student: Walker Wright-Moore (BS), assisted with induced spawning experiments for southern flounder as well as with nutrition experiments with black sea bass. Results will be incorporated into a MS Thesis in Marine Science. Graduate student: Andrew D. Ostrowski, assisted with red porgy larviculture and live prey enrichment studies. Undergraduate students: T. Gibson, R.Doyle, A. Markwith, C.Kelly, assisted with experiments. TARGET AUDIENCES: Target audiences included the graduate and undergraduate students listed above who participated in the experimental work and/or derived MS thesis projects, commercial collaborators on southern flounder and black sea bass culture who benefited from the culture technologies developed during this project, and the general aquaculture community, including researchers, commercial farmers, educators and public policy makers to whom project results were disseminated through conference presentations. Efforts included aquaculture practicum experiences and experimential learning opportunities for graduate students and interns at UNCW, extension and outreach of technical advances to private collaborators associated with this project and through dissemination of project results through publications. Efforts include acts or processes that deliver science-based knowledge to people through formal or informal educational programs. Examples include: formal classroom instruction, laboratory instruction, or practicum experiences; development of curriculum or innovative teaching methodologies; internships; workshops; experiential learning opportunities; extension and outreach. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
1. Standardize hormone therapies for induced breeding of F1southern flounder. In terms of spawning performance (larval production), there were significant effects of hormone dose, with maximum production observed at the low dose (5 ug/kg bw) for both LHRHa and sGnRHa. Larval production was higher with LHRHa in F1 females and with sGnRHa in wild females. 2. Reduce southern flounder fingerling production costs by minimizing reliance on live prey through developing artificial microdiets. Expt. 1 indicated that a MBD containing menhaden, squid, krill, herring meal and attractants could be used to reduce live feed cost. Expt. 2 showed that abrupt weaning to MBD beginning on d11dph resulted in poor survival compared to larvae co-fed with MBD or fed live feed only. However, co-feeding MBD with one half live feed rations from d11ph produced no difference in survival compared to feeding live feed alone. 3. Evaluate substitution limits of underutilized plant and animal by-products as a fish meal replacement in diets. 3.1. Replacement of menhaden fish meal by solvent extracted soybean meal supplemented with amino acids. The optimum level of fish meal protein replacement with solvent extracted soybean protein was 70% with 7.5% squid meal and 5% krill meal and with or without supplemental methionine and lysine in the diet. 3.2. Replacement of menhaden fish meal by poultry by-products and fermented poultry by-products. Menhaden fish meal protein could be replaced up to 70% by soybean protein in juvenile diets when supplemented with small amount of squid meal, krill meal, fermented poultry by products or methionine and lysine without impairing growth. Poultry by-product meal is a promising alternative protein substitute for fish meal protein for sustainable diets. 3.3. Replacement of menhaden fish meal by meat and bone meal. Fish meal protein may be replaced by soybean meal protein (without supplemental Met and Lys) at levels up to 70% in the diet. Fish meal replacement by meat-and-bone meal and poultry-by-product meal at levels of at least 30% and 60%, respectively, were successful. 3.4. Formulation of different cost effective diets using a combination of different alternative protein sources (e.g. soybean meal, poultry by-product, and meat and bone meal) and their effects on growth and body composition. The optimal diet producing no significant reduction in performance contained only 6% FM (70% fish meal protein replacement) in combination with soybean meal, poultry by-product meal, meat and bone meal, squid meal, and krill meal. A practical low fish meal diet containing a mixture of alternative protein sources is possible for juvenile black sea bass. 4. Determine the effects of these feeds on fish biochemical composition (In progress). 5. Optimize enrichment methods for live feeds for larval red porgy. Performance was better in the Algamac treatment and the DC DHA Selco treatment than in the control. Survival was significantly higher in the DHA Selco treatment than in Algamac 3000 and the control. Prey enrichment protocols have been established for red porgy that will generate more cost-effective juvenile production output.

Publications

  • Alam, M.S., Watanabe W.O., Sullivan, K.B. 2008. Recent progress on development of nutritionally-balanced, cost-effective and environmentally-friendly diets for the culture of black sea bass Centropristis striata and southern flounder Paralichthys lethostigma. Eighth Joint Meeting of the Seafood Science and Technology Society and Atlantic Fisheries Technology Conference in conjunction with the 1st NC Marine Biotechnology Symposium 2008, Book of Abstracts. Wrightsville Beach, NC, Oct 20-23, 2008.
  • Alam, M.S., W.O. Watanabe, K.B. Sullivan and W. D. Wright-Moore. 2010. Partial and full replacement of menhaden fish meal protein by soybean meal protein supplemented with or without amino acids and attractants in the diets of juvenile black sea bass Centropristis striata. Book of Abstracts, Aquaculture America 2009, Seattle, WA, USA
  • Dawson, M.R., M.S. Alam, and W.O. Watanabe. 2010. Effects of poultry by-product meal protein substitution for fish meal protein on growth performance, body composition, and digestibility of juvenile black sea bass Centropristis striata. Book of Abstracts, World Aquaculture 2010, San Diego, California.
  • Alam, M.S., Watanabe, W., Sullivan, K.B., Rezek, T., Seaton, P. 2011. Replacement of menhaden fish meal protein by solvent extracted soybean meal protein in the diet of juvenile black sea bass Centropristis striata supplemented with or without squid meal, krill meal, methionine and lysine. Submitted to North American Journal of Aquaculture (In review).
  • Alam, M.S., W.O. Watanabe, Myers, A.R. 2011. Growth performance and survival of larval southern flounder Paralichthys lethostigma fed commercial microdiets or formulated zein bound microdiets containing different protein sources. In preparation.
  • Sullivan, K.B. M.S. Alam, and W.O. Watanabe. 2008. Replacement of fish meal by soybean meal, poultry byproduct meal and meat and bone meal in diets for juvenile black sea bass Centropristis striata. Book of Abstracts, Aquaculture America 2008, Orlando, Florida
  • M. S. Alam, W.O. Watanabe, H.V. Daniels, P.R. Ferket, A. Gill, T.C. Rezek, A.R. Myers and M.R. Dawson. 2011. Replacement of fish meal protein by soybean meal protein supplemented with or without fermented poultry by products, squid meal, krill meal and amino acids in the diet of juvenile black sea bass Centropristis striata. Book of Abstracts, Aquaculture America 2011, New Orleans, Louisiana.
  • Dawson, M.R., Alam, M.S. and W.O. Watanabe. 2011. Replacement of fish meal protein by a combination of alternative protein sources in practical diets for juvenile black sea bass Centropristis striata. Book of Abstracts, Aqua Amercia 2011, New Orleans, Louisiana.


Progress 08/01/08 to 07/31/09

Outputs
OUTPUTS: Objective 1. Hormone-induced breeding of first-generation (F1) southern flounder (SF). SF females (33 F1 + 17 wild) were treated with pelleted LHRHa at different doses (5, 25 and 50 microgram/kg), and 78% were successfully strip-spawned. Dose effects on egg production and fertilization and hatching success were determined. Objective 2. Substitution limits of plant and animal by-products as a fish meal (FM) replacement in black sea bass (BSB) diets. A series of eight 6-12 wk experiments determined the effects of replacing menhaden FM with other protein sources. Initial fish weights ranged from 5-36 g. Expt. 2.1. Seven diets replaced FM protein with solvent-extracted soybean meal (SM) protein at 0, 10, 20, 30, 40, 50 and 60%, without methionine (met) and lysine (lys), but with 1% attractants (taurine, betaine, alanine and glycine). Expt. 2.2. Six diets replaced FM protein with SM protein at 0, 60, 70, 80, 90 and 100%. A diet with 60% of the FM replaced with poultry-by product meal (PBM), and a diet with 30% of the FM replaced with meat-and-bone meal (MBM) were also tested. All diets contained 1% attractants, but no met or lys. Expt. 2.3. Seven diets replaced FM protein with SM protein at 0, 50, 60, 70, 80, 90 and 100%, without met, lys, and attractants, but all diets contained 7.5% squid meal (SqM) and 5% krill meal (KM). Expt. 2.4. Six diets replaced FM protein with SM protein at 0, 40, 50, 60, 70 and 80% without supplemental met, lys, attractants, SqM and KM. Expt 2.5. Five diets replaced FM protein with SM protein at 0, 60, 70 and 80 and 90% without met, lys, SqM, KM and attractants. Three diets replaced FM protein by SM protein at 60, 70, and 80% with supplemental met and lys, but without SqM, KM and attractants. Expt. 2.6. Three diets were tested at a pilot scale: Diet 1 (control) contained 50% menhaden FM; Diet 2 replaced 70% menhaden FM protein with SM protein; and Diet 3 combined FM, SM, PM and MBM. Expt. 2.7. Eight diets incorporated different levels of PBM to replace FM protein, without supplemental amino acids, SqM, KM and attractants. Expt. 2.8. Eight diets were formulated with a combination of different protein sources, including FM, SM, SqM, KM, PBM and MBM. Expt. 8 determined the maximum replacement of FM protein by PBM protein and by MBM protein (individually or in combination) in diets containing a high level of SM with and without SqM and KM. All diets had 43% CP and 12% CL. Objective 3. Optimize live prey enrichment methods for larval red porgy. Expt.3.1. First feeding larvae (d3ph) were fed rotifers enriched with one of four commercial diets (Rotifer Diet, Protein Selco, Algamac 3000, and Algamac 3000/ARA mix) and under two feeding regimes (1x or 2x day). Expt. 3.2 The effects of commercial enrichment products products (DC DHA Selco and Algamac 3000) on the performance of older larvae (d13ph) fed artemia were compared. PARTICIPANTS: Individuals: Project Director: Dr. Wade O. Watanabe, served as project coordinator and technical and scientific supervisor, including project administration. Co-Principal Investigator: Dr. Md. Shah Alam: provided technical and scientific supervision for nutrition experiments on alternative protein sources to fish meal in black sea bass diets. Research Associate: Patrick M. Carroll; provided operational assistance on experimental work through facilities design, setup, and maintenance. Research Associate: Troy C. Rezek (MS), provided technical assistance for induced spawning experiments of southern flounder using LHRHa and on larviculture experiments with red porgy. Partner Organizations: North Carolina Sea Grant, provided funding support which help leverage induced spawning experiments with southern flounder NOAA Marine Aquaculture Program, provided funding support which helped leverage nutrition experiments with black sea bass Golden Leaf Foundation, provided funding support which helped leverage induced spawning experiments with southern flounder. Collaborators and contacts: Dr. James Morris, Center for Coastal Fisheries and Habitat Research (CCFHR), National Ocean Service (NOS), NOAA, provided red porgy embryos which were used for larval rearing experiments on live prey enrichment. Dr. Harry V. Daniels (North Carolina State University), received southern flounder embryos that were produced in induced spawning experiments. These embryos were used for experimental purposes. Mr. Keith Hairr (Carolina Flounder LLC), received southern flounder embryos produced in induced spawning experiments. These embryos were used for commercial hatchery purposes at CF. Mr. Shawn Longfellow (Blue Ocean Farms LLC), received southern flounder fingerlings from CF that were produced in induced spawning experiments. These fingerlings were used for commercial growout and marketing at BOF. Mr. Ted Davis (Aqua Plantations LLC), received black sea bass fingerlings from UNCW that were produced in nutrition experiments. These fingerlings were used for commercial growout and marketing purposes at AP. Training or professional development: Graduate student: Katherine B. Sullivan (MS), conducted experiments on replacement of fish meal in black sea bass diets with soybean meal, poultry meal and meat and bone meal. Results were incorporated into a MS Thesis in Marine Science. MS degree was received in Fall 2008. Graduate student: Matthew R. Dawson (BS): conducted experiments on replacement of fish meal in black sea bass diets with poultry meal and combined alternative protein sources. Results will be incorporated into a MS Thesis in Marine Science. Graduate student: Walker Wright-Moore (BS), assisted with induced spawning experiments for southern flounder as well as with nutrition experiments with black sea bass. Results will be incorporated into a MS Thesis in Marine Science. Graduate student: Andrew D. Ostrowski, assisted with red porgy larviculture and live prey enrichment studies. Undergraduate students: Tyler Gibson, Richard Doyle, Annie Markwith, Cassandra Kelly, assisted with experimental work. TARGET AUDIENCES: Target audiences included the undergraduate and graduate students listed above who participated in the experimental work and/or derived undergraduate credit hours and MS degrees and thesis projects, commercial collaborators on southern flounder and black sea bass aquaculture who benefited from the culture technologies developed during this project, and the general aquaculture community, including researchers, commercial farmers, educators and public policy makers to whom project results were disseminated through conference presentations and publications. Efforts included aquaculture practicum experiences and experimential learning opportunities for graduate students and interns at UNCW, extension and outreach of technical advances to private collaborators associated with this projec and through dissemination of project results through publications. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
Objective 1: Results to date suggested that in F1 females, egg production and quality were maximized at the lowest hormone dose rate of 5 microgram/kg LHRHa slow release pellet. However, in wild-caught females, egg production and egg quality were maximized at an intermediate dose of 25 microgram/kg LHRHa. The results further suggested that a high dose of 50 microgram/kg (typically used for induced spawning of SF) can over stimulate both F1 and wild flounder. Methods for reliable spawning of F1 SF females will help to establish long-term selective breeding programs for domestication of SF as an aquacultured species. Objective 2: Menhaden FM protein could be replaced at a level of 70% by solvent extracted SM protein with 7.5% SqM, 5% KM and 1% attractants in the diets of juvenile BSB, but not more than 60% without attractants. This maximum replacement level was not significantly affected by supplementing met and lys in a SM based diet. FM replacement by MBM and PBM at levels of at least 30% and 60%, respectively, were also successful. Higher substitution levels are now being tested. BSB were able to utilize a high level of SM in substitution of FM. Pilot scale growout studies also indicated that BSB were able to utilize a high level of SM in replacement of FM. All of these results indicated that a cost-effective alternative protein based diet formulation is possible for BSB without compromising growth. Reduced growth at more than 70% substitution by SM protein may be attributed to a higher linoleic acid content, a lower level of met or lys, or anti-nutritional factors in the SM protein. In addition, PBM protein was substituted for FM protein at levels as high as 80%, with no reduction of growth performance. Based on growth and survival, the optimal diet producing no reduction in performance contained only 6% FM (70% FM protein replacement) in combination with SBM, PBM, MBM, SM, and KM. A short term laboratory study using combinations of alternative protein sources indicated that SM, PBM, and MBM are promising alternative protein substitutes for FM protein for sustainable diet development in BSB. Objective 3: This study determined the effects of commercially available enrichment products for rotifers and artemia and of different feeding regimes on performance of early larvae from yolk-sac through metamorphic stages. Expt.3.1 No differences in the effects of these enrichment product on growth, survival, or hypersaline stress resistance were observed. Expt. 3.2. By d33ph, growth and survival were higher in larvae from the Algamac 3000 treatment and the DC DHA Selco treatment than in the control. Optimal prey enrichment protocols for RP using state of the art products and techniques will generate more cost-effective juvenile production output.

Publications

  • Paper entitled "Effects of poultry by-product meal protein substitution for fish meal protein on growth performance, body composition, and digestibility of juvenile black sea bass Centropristis striata" by M.R. Dawson, M.S. Alam, and W.O. Watanabe, presented at World Aquaculture 2010, March 2010, San Diego, CA.
  • Paper entitled "Partial and full replacement of menhaden fish meal protein by soybean meal protein supplemented with or without amino acids and attractants in the diets of juvenile black sea bass Centropristis striata, presented by M.S.Alam, W.O. Watanabe, K.B. Sullivan and W.D. Wright-Moore presented at Aqua America 2009, February 2009, Seattle, WA.
  • Watanabe, W. O., and Alam, M. S. 2010. Plant-based feeds for black sea bass show promise. NOAA-Aqua Feeds initiative. Case study Two. Aqua Feeds Initiative: 19-20.