Progress 07/15/08 to 07/14/11
Outputs
OUTPUTS: Striped Bass Egg Quality - Ovaries of female striped bass producing poor quality eggs had a dysfunctional transcriptome before the breeding season. Results of these experiments were shared with aquaculture science and industry stakeholders at the 2008, 2009, and 2010 International Plant and Animal Genome Conference, the 2008, 2009, and 2010 Annual Meeting of the Striped Bass Growers Association at the Aquaculture America Conference, and at the 2008, 2009, and 2010 North Carolina Aquaculture Development Conference. These findings also were shared with producers of striped bass fingerlings and with the staff of two USDA-ARS Laboratories. All of the high-quality EST sequences used to construct the striped bass ovarian transcriptome were posted to the NCBI Short Read Archive (GenBank: SRX007394), the contiguous gene sequences making up the transcriptome were posted on the U.S. National Animal Genome Project website, the high definition Agilent microarray (eArray Group: Striper Group, Design ID: 029004) is available from Agilent. Feed Intake and Growth Regulation; We have cloned the gene for striped bass NPY, a presumed orexigenic hormone, characterized its spatial distribution within the brain of hybrid striped bass (HSB). NPY mRNA expression in the different brain regions were measured in animals during short-term and long-term fasting and refeeding. We also cloned and sequenced leptin, showed it to be distributed primarly in the liver of HSB, and assessed its effect on appetite and regulation during different catabolic and anabolic states. The research was disseminated at the World Aquaculture Society, Society of Integrative and Comparative Biology, and North American Society of Comparative Endocrinology, and the annual North Carolina Aquaculture Conference. The technologies for inducing compensatory growth and improving feed efficiency of HSB have been tested in ponds with an HSB grower cooperator (Pungo Fisheries). Hybrid Striped Bass Pond Effluents: Studies of water conservation practices have shown dramatic changes in effluent quality that would lead to compliance with regulatory standards but are economically not viable. Evaluations of chemical treatments have shown that effective concentrations are cost prohibitive for use on commercial farms. Retaining water in ponds post-harvest is the most economical means of reducing effluents and minimizing waste production. These findings have been presented to producer groups and the Annual North Carolina Aquaculture Development Conferences2008, 2009, and 2010). Water Reuse Technology: Seventy five individuals that attended the aquaculture effluents workshop at the 2010 NC Aquaculture Development Conference gained knowledge about the efforts ongoing to solve current and future issues in freshwater and marine aquaculture effluents. Results from this project were included in materials presented in two day short-courses entitled Aquaculture Recirculating Technology presented in Brisbane, Australia, in 2008 and Hobart Australia in May 2010. Additionally, an update of the results of this project was presented at the annual NC Aquaculture Development Conference in February, 2011. PARTICIPANTS: Russell Borski, Department of Biology, NC State (Co-Principal Investigator); Eugene Won, PhD graduate research assistant, NC State; Andrew Baltzegar, PhD student, NC State; Nishika Patel, Undergraduate student, NC State; Emily Brune, Undergraduate student, NC State; Brad Ring, Aquaculture Technician, NC State University; John Davis, Aquaculture Technician, NC State University; Larry Riley, Associate Professor, California State University-Fresno. Mr. Steve Locke, Owner, Castle Hayne Fisheries; Dr. Ohla Sydorovych, Department of Agricultural and Resource Economics; Mr. Brian Shannon, Research Technician, NC State University. TARGET AUDIENCES: This research targeted growth physiologists and endocrinologists, genomic and proteomic scientists, aquaculture sciences, and the aquaculture industry. Results of this work have been disseminated to producers and scientists through various mechanisms that include presentations at conferences and universities. The research also contributed to the research experiences of two undergraduate students majoring in biochemistry or interested in aquaculture. The National Hybrid Striped Bass Grower's Association will directly benefit from this work. The PI's also teach graduate course in endocrinology and aquaculture for which some of the research outlined here is used in classroom instruction. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Striped Bass Egg Quality - Our findings illustrate that subtle, collective changes in expression of a limited suite of genes representing only 2% of the queried ovary transcriptome explains most of the variation in egg quality. Gene expression patterns in ovaries sampled before spawning explained over 90% of the eventual variance in embryo survival and, when transcriptome data was input from new fish, the trained ANN model could predict their egg quality with ~80% accuracy. A correlation analysis of gene expression indicates that the collective dysfunction of the ubiquitin-26S proteasome, COP9 signalosome, and subsequent control of the cell cycle may be the primary cause of embryonic developmental incompetence in striped bass eggs. The combined transcriptional differences exhibited the most powerful molecular relationship to this biologically complex trait ever described and suggest that the oocyte transcriptome is predictive of egg quality. Feed Intake and Growth Regulation: We have cloned the gene for NPY and established the spatial distribution of the hormone in an important finfish cultivar. We demonstrate the gene for NPY is upregulated over 3 days in the hypothalamus and is increased in the telencephalon over longer term (6 - 24 day) fasting. The NPY increase during fasting suggests the hormone may enhance appeitite, priming fish to undergo a hyperphagic response during periods of refeeding. We also show that leptin declines during fasting, increases during anabolic states and stimulates appetite. This indicates the hormone promotes anabolic processes and inhibits appetite to maintain energy homeostasis. Hybrid Striped Bass Pond Effluents: Results from this work have been subjected to a thorough cost-benefit analysis and have been published in a peer-reviewed journal. Water Reuse Technology: More than 95% of the suspended solids entering the geotextile bag were removed from the waste stream. COD reduction varied from 18% - 85%. Total nitrogen reduction varied from 42% to 96% and Total phosphorus reduction ranged from a low of 18% to a high of approximately 67%. The operation of the two different secondary and tertiary marine waste treatment systems showed clear differences in operational capabilities that varied with the seasons. The moving bed / fixed bed filter combination provided year-round nitrogen removal; however, the fixed bed filter required organic carbon additions for adequate denitrification activity. The constructed wetlands provided adequate nitrogen removal during the warmer growing season, yet did not provide adequate nitrogen removal in the winter months. Phosphorus was removed year round with chemical precipitation in both the biochemical and constructed wetlands systems. The impact of this research points the way to a hybrid system combining a moving bed / fixed bed system with a free-water constructed wetland for additional denitrification and final polishing. Either system would be adequate for seasonal treatment (pond culture systems) while the hybrid system would be required for a year round system (indoor recirculating system).
Publications
- Sydorovych, O. and H. Daniels. 2011. Economic Analysis of Alternative Effluent Treatment Options for Pond Production of Hybrid Striped Bass in Aurora, North Carolina. Aquaculture Economics & Management. 15:46-70.
- Picha, M.E., Strom, C.N., Riley, L.G., Walker, A.A., Won, E.T. Johnstone, W.M. and Borski, R.J. (2009). Plasma ghrelin and growth hormone regulation in response to metabolic state in hybrid striped bass: Effects of feeding, ghrelin and insulin-like growth factor-I on in vivo and in vitro GH secretion. General and Comparative Endocrinology. 161:365-372.
- Won, E.T., Baltzegar, D.A., Picha, M.E., and Borski, R.J. 2009. Cloning and regulation of hepatic leptin mRNA expression by nutritional status in hybrid striped bass (Genus Morone). Integrative and Comparative Biology. 49:E184-E184.
- Borski RJ, Won ET and Baltzegar DA (2011). Leptin stimulates hepatic growth hormone receptor expression: possible role in enhancing gh-mediated anabolic processes in fish. Front. Endocrin. Conference Abstract: NASCE 2011: The inaugural meeting of the North American Society for Comparative Endocrinology. doi: 10.3389/conf.fendo.2011.04.00111
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Progress 07/15/09 to 07/14/10
Outputs
OUTPUTS: Feed Intake and Growth Regulation: We have cloned the gene for striped bass NPY, and characterized its spatial distribution within the hypothalamus and brain of hybrid striped bass (HSB) by in situ hybridization. NPY gene expression occurs throughout the brain and we found that its mRNA levels within the whole brain are not regulated by nutritional state. Studies are now underway to isolate the hypothalamus from the remainder of the brain to evaluate the influence of nutritional state on NPY mRNA in the putative feeding center only. Striped Bass Egg Quality: We have found that atresia could be effectively delayed for prolonged periods in spring by holding broodfish at low temperature (12 degrees centigrade) but that, even with atresia controlled, poor egg quality persisted in about half of the striped bass. Experiments were undertaken to discover the extent to which profiles of activation of specific ovarian genes might be a proximal cause of poor egg quality. Females yielding good or bad quality spawns were biopsied for oocytes before breeding activities and the biopsies were extracted for mRNA which was subjected to microarry to measure expression of ~15,000 ovary/egg genes in the samples and to compare gene expression patterns to resulting hatchery measures of egg quality. Results of these experiments were shared with graduate students and postdoctoral researchers working in aquaculture at NCSU and with science and industry stakeholders in formal presentations and informal discussions at the 2010 International Plant and Animal Genome Conference, the Annual Workshop on Genetic Improvement and Selective Breeding for the Hybrid Striped Bass Industry, and the North Carolina Aquaculture Development Conference. Hybrid Striped Bass Pond Effluents: Studies of water conservation practices have shown dramatic changes in effluent quality that would lead to compliance with regulatory standards but are economically not viable. Evaluations of chemical treatments have shown that effective concentrations are cost prohibitive for use on commercial farms. Retaining water in ponds post harvest is the most economical means of reducing effluents and minimizing waste production. These findings have been presented to producer groups and the Annual North Carolina Aquaculture Development Conference. Water Reuse Technology: Mr. Todd Guerdat is conducting PhD research at a new Research Center near Marshallberg, NC. The experimental setup consists of a state of the art saltwater aquaculture waste research system to compare biological and chemical methods for treating aquaculture waste to the capabilities of constructed wetlands. These systems have been operational for 9 months. During the reporting period, Dr. Losordo served as chair or a member of 4 graduate research committees. PARTICIPANTS: Russell Borski, Department of Biology, NC State (Co-Principal Investigator); Eugene Won, PhD graduate research assistant, NC State; Andrew Baltzegar, PhD student, NC State; Nishika Patel, Undergraduate student, NC State; Emily Brune, Undergraduate student, NC State; Brad Ring, Aquaculture Technician, NC State University; John Davis, Aquaculture Technician, NC State University; Larry Riley, Associate Professor, California State University-Fresno. Mr. Steve Locke, Owner, Castle Hayne Fisheries; Dr. Ohla Sydorovych, Department of Agricultural and Resource Economics; Mr. Brian Shannon, Research Technician, NC State University. TARGET AUDIENCES: This research targeted growth physiologists and endocrinologists, genomic and proteomic scientists, aquaculture sciences, and the aquaculture industry. Results of this work have been disseminated to producers and scientists through various mechanisms that include presentations at conferences and universities. The research also contributed to the research experiences of two undergraduate students majoring in biochemistry or interested in aquaculture. The National Hybrid Striped Bass Grower's Association will directly benefit from this work. The PI's also teach graduate course in endocrinology and aquaculture for which some of the research outlined here is used in classroom instruction. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Feed Intake and Growth Regulation: We have cloned the gene for NPY and established the spatial distribution of the hormone which is thought to be critical to appetite regulation and to other behavioral processes in an important finfish cultivar. We have also established the dose dependent effects of ghrelin and leptin on feeding behavior in the HSB. Striped Bass Egg Quality: We discovered that losses in bad spawns occurred in the first 4 h after fertilization (up to mid-blastula stage), when the embryo is totally dependent on maternal transcripts. Data analysis using artificial neural networks (ANNS) and machine learning tools produced models of the relation of transcriptome to egg quality revealing that expression of as few as 250 genes can explain over 95% of variation in egg quality and that, when data from new fish were input, the models could predict egg quality with ~ 80% accuracy.These are the most compelling data that maternal transcripts are a major determinant of egg quality in farmed fishes and these finding have led to our establishment of a broad consortium of scientists who will explore the significance of these findings in temperate basses, rainbow trout and other teleosts. Hybrid Striped Bass Pond Effluents: Results from this work have been subjected to a thorough cost-benefit analysis and will be published in a peer-reviewed journal. Water Reuse Technology: Results from this project were included in materials presented in a two day short-course entitled Aquaculture Recirculating Technology presented in Hobart Australia in May 2010. Both projects were described in two presentations at the Austral-Asian Aquaculture Conference in Hobart Australia in May 2010. Additionally, the results to date were described in the research section of the North Carolina Aquaculture Development Conference in January 2010 in Atlantic Beach, NC. More than 20 groups or individuals were provided tours of the aquaculture facilities at NC State as part of this project. The results from this research project are extended to the business community via direct consultation with the agribusiness community in North Carolina. As part of our outreach effort, Dr. Losordo and Mr. Delong answered hundreds of telephone, email and in-person requests for information related to this project.
Publications
- No publications reported this period
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Progress 07/15/08 to 07/14/09
Outputs
OUTPUTS: Marine Effluents: Construction of the wastewater treatment facilities located behind the Marine Aquaculture Research Center (MARC) began June 1, 2009. Since then, four primary systems have been constructed: Solid waste treatment (Geotube), Solids-free wastewater storage, constructed treatment wetlands, and the biological/chemical wastewater treatment system. The design, operation, and maintenance of these systems are all part of a PhD research project conducted by Todd Guerdat of the Biological and Agricultural Engineering department at North Carolina State University. PARTICIPANTS: Marine Aquaculture Effluents: Thomas M. Losordo, Professor, Department of Biological Engineering, NC State; Todd C. Guerdat, Graduate Research Assistant, Department of Biological and Agricultural Engineering, NC State; Marc Turano, Aquaculture Specialist, NC Sea Grant Program; Nick Reynolds, Aquaculture Technician, Department of Biological and Agricultural Engineering, NC State; Brett Harrison, Research Technician, NOAA Labs, Beaufort, NC. TARGET AUDIENCES: Marine Aquaculture Effluents: The target audience for this project are the citizens of North Carolina with interest in the development of aquaculture activities with marine species in coastal North Carolina. Seventy five individuals that attended the aquaculture effluents workshop at the 2010 NC Aquaculture Development Conference gained knowledge about the efforts ongoing to solve current and future issues in freshwater and marine aquaculture effluents. PROJECT MODIFICATIONS: Marine Aquaculture Effluents: The only modification since writing this proposal is that we are focusing exclusively on the problem at the Marine Aquaculture Research Center in coastal NC. We will not operate a marine geotube at the NC State Fish Barn due to changes in the project at that site.
Impacts
Marine Aquaculture Effluents: Mr. Guerdat described the facilities and their potential for treating marine aquaculture effluents at a workshop at the 2010 NC Aquaculture Development Conference.
Publications
- No publications reported this period
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