Progress 06/15/10 to 06/14/13
Outputs
Target Audience:Aquaculture producers in North Carolina as well as the broader U.S. aquaculture industry. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Graduate student training for three PhD students. Additional training for one postdoctoral researcher. How have the results been disseminated to communities of interest?Our results have been presented at annual producers meetings in NC. and at scientific meetings of the World Aquaculture Society, the U.S. Chapter of the World Aquaculture Society (Aquaculture America), and at state meetings of the NC Water Resources Research Institute. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
We have made significant advances in our understanding of how egg development is regulated and can be controlled for the purposes of improving spawning perfomance in domesticated farm-raised fish. Our insights into the genetic and proteomic processes that govern the development of high quality eggs have ultimately led to the near elimination of the use of hormones to control spawning and approximatley 40% improvement in spawning success of brood fish. Land-application of fish farm effluents appears to be a feasible means of disposing of these effluents while improving their quality to meet strict regulatory limits. This system is unique and innovative and appears to offer additional ecosytem services to producers with available land adjacent to their farms and would serve as part of a larger effluent management strategy. Indoor production of hybrid striped bass is now feasible for all phases of production after the initital hatchery phase. Time to harvest is dramatically reduced with the use of a recirculating aquaculture system (RAS) which translates to shorter production times and increased efficiency of production.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2014
Citation:
Chapman*, R.W., Reading*, B.J., and Sullivan*, C.V. 2014. Ovary transcriptome profiling via artificial intelligence reveals a transcriptomic fingerprint predicting egg quality in striped bass, Morone saxatilis. PLoS ONE 9(5):e96818.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2014
Citation:
Williams, V.N., Reading, B.J., Hiramatsu, N., Amano, H., Glassbrook, N., Islam Williams, T., and Sullivan, C.V. 2014. Multiple vitellogenins and product yolk proteins in striped bass, Morone saxatilis: molecular characterization and processing during oocyte growth and maturation. Fish Physiology and Biochemistry 40(2):395-415.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2013
Citation:
Reading, B.J., Williams, V.N., Chapman, R.W., Islam Williams, T., and Sullivan, C.V. 2013. Dynamics of the striped bass (Morone saxatilis) ovary proteome reveal a complex network of the translasome. Journal of Proteome Research 12:1691-1699.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2012
Citation:
Reading, B.J., Chapman, R.W., Schaff, J.E., Scholl, E.H., Opperman, C.H., and Sullivan, C.V. 2012. An ovary transcriptome for all maturational stages of the striped bass (Morone saxatilis), a highly advanced perciform fish. BMC Research Notes 5:111 (doi: 10.1186/1756-0500-5-111).
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Progress 06/15/11 to 06/14/12
Outputs
OUTPUTS: Accumulation by growing oocytes of the three forms of yolk precursor protein (vitellognin, Vtg) was evaluated by in striped bass by isotope-free quantitative tandem mass spectrometry (MS/MS), which revealed that maximum accumulation of VtgAb and VtgAa occur during early- and mid-vitellogenesis, respectively, while VtgC accumulates throughout oocyte growth. We also discovered a new class of Vtg receptor (Vtgr), named LRX+1 based on its domain structure, obtained the full-length cDNA coding sequence for the new receptor, and verified that the receptor mRNA is mainly expressed in ovary and localized to the ooplasm of previtellogenic oocytes. The MS/MS revealed peak expression of LRX+1 protein in ovary of bass during late-vitellogenesis. These results were reported at the 9th International Symposium on Reproductive Physiology of Fish. Our analyses of blood and mucus samples collected on a monthly basis from individually-tagged pubescent Atlantic sturgeon for measurement of sex steroid levels and other indicators of gender and maturation also was completed. At the end of the sample period, we determined gender of the fish using gonadal biopsy. We also examined the fish for discriminatory secondary characteristics of gender. Results for Atlantic sturgeon suggest that while plasma steroids can be predictive of gender, mucus samples did not correlate well with plasma steroid levels. Similar evaluations of Russian sturgeon were initiated and are continuing. These findings were reported at the 2012 North Carolina Aquaculture Development Conference. In studies conducted to evaluate how stress adversely affects the growth of farmed hybrid striped bass (HSB), we completed a growth trial to induce catabolic, normal anabolic and superanabolic states following alterations in feeding regimes. This followed establishment of radioreceptor assays to characterize binding characteristics of the intracellular and putative membrane receptor for cortisol in fish; and validation an extraction and radioimmunoassay for measurement of cortisol in blood and tissue. The studies have provided training on conducting growth trials and on biochemical and endocrinological techniques for measuring levels of a stress steroid and its tissue sensitivity in fish to two graduate and two undergraduate students. The research was disseminated to the scientific community at the International Symposium on Fish Endocrinology and the North American Society of Comparative Endocrinology. The economic feasibility of disposal of HSB farm pond effluents via land application was evaluated by using water from eighteen research ponds stocked with HSB to irrigate two plots of land planted with various tree species. Pond water was applied to the plots until standing water was observed and data on daily irrigation volumes, percolation rates, and tree transpiration rates was collected year round. The results of this work have been presented to HSB producer groups, at the annual N.C. Aquaculture Development Conference and in one-on-one meetings during numerous farm visits and with state regulatory personnel. PARTICIPANTS: Prof. Craig Sullivan, Dept. Biology, NC State University (NCSU), Principal Investigator (PI); Prof. Harry Daniels, Dept. Biology, NCSU, Co-PI; Prof. Jeff Hinshaw, Dept. Biology, NCSU, Co-PI, Prof. Russell Borski, Dept. Biology, NCSU, Co-PI; Prof. Tom Losordo, Dept. Biological and Agricultural Engineering, NCSU, Co-PI; Dr. Andy McGinty, Superintendent, NCSU Pamlico Aquaculture Field Laboratory; Dr. Benjamin Reading, Postdoctoral Research Associate, Dept. Biology, NCSU; Valerie Williams, Ph.D. graduate student, Dept. Biology, NCSU; Jamie Mankiewicz, M.S. graduate student, Dept. Biology, NCSU; Miller Johnstone, PhD graduate research assistant, Dept. Biology, NCSU; Andrew Baltzegar, PhD graduate student, Dept. Biology, NCSU; Jonathon Douros, PhD graduate student, Dept. Biol. NCSU; Everett Wilson, undergraduate student, NCSU; Jay Patel, undergraduate student, NCSU; Christina Shenton, M.S. graduate student, Dept. Biology, NCSU; Katelyn Molloy, Undergraduate student, Dept. Biology, NCSU; Adriane Gill, Ph.D. graduate Student, Dept. Biology, NCSU; Brad Ring, Research Specialist, Dept. Biology, NCSU; Tyler Nethers, Research Technician, Dept. Biology, NCSU; Michael Hopper, Research Specialist, NCSU Pamlico Aquaculture Field Laboratory; John Davis, Research Technician, Dept. Biology, NCSU. TARGET AUDIENCES: This research targeted reproductive and growth physiologists and endocrinologists, aquaculture scientists including engineers, aquaculture industry stakeholders including include Striped Bass Growers Association members, especially producers of hybrid striped bass in eastern North Carolina, the U.S. aquaculture community in general, and government regulators of water resources and water quality. Also targeted were graduate an undergraduate students involved in aquaculture research or education. The project PI and Co-PIs are also engaged in classroom instruction in courses such as Fish Physiology, Aquaculture, and Endocrinology for which some of the lines of research reported here is used in formal lectures in the classroom. Other efforts at reaching these audiences included presentations at the 9th International Symposium on Reproductive Physiology of Fish and at the NC Aquaculture Development Conference, as well as direct contact with North Carolina HSB growers while conducting research or assisting with production activities on their farms. This research provided for the instruction of a postdoctoral Research Associate, and several graduate and undergraduate students via hands on training during execution of their research. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
In our studies of egg quality in striped bass, the detailed characterization of when the different forms of Vtg are accumulated as yolk proteins, and our discovery of a new form of Vtg receptor, enhance our knowledge of the molecular details of oocyte growth in the striped bass and its relatives and set the stage for their evaluation in females producing good versus poor quality eggs. A post-doctoral research scholar, a doctoral student, and an undergraduate student were trained in advanced proteomics methodologies for reproductive biology in this research. In our studies of stress and growth, we generated animals that show low, average and elevated growth rates characteristic of catabolic, normal anabolic and superanabolic (compensatory growth) states. These data demonstrate that HSB can undergo compensatory growth characterized by improved feed efficiency once they are alleviated of a period of fasting. Assays for measures of a key hormone indicative of stress responses in fish, cortisol, were produced, and bioassays for measuring the classical intracellular abundance and affinity of cortisol receptor were established. These developments allow assessment of environmental parameters that produce stress and that can subsequently impair growth potential, sex determination and osmotic responses of aquacultured fish. Sturgeon farming is a new aquaculture enterprise adding to the diversity of aquaculture products from North Carolina. Since only the females should be kept until they are fully mature, a method of determining the sex of the animals at the earliest possible age is needed to make the culture of sturgeon efficient. The currently used techniques are invasive, expensive, and can't be reliably performed until the animals are at least two to three years old. We are continuing to explore non-invasive and minimally-invasive techniques for sex determination in Russian sturgeon and other sturgeon species that can be employed in a commercial aquaculture setting. North Carolina State regulators in the Division of Water Quality are monitoring the results of the research on land application of HSB pond effluents and the final results will be used to help establishBest Management Practices for HSB pond culture in North Carolina; this land-application is a novel approach to the treatment of aquaculture effluents. The establishment of this research tree plot provides us with a unique capability to evaluate these systems for the removal of environmental contaminants from fish production.
Publications
- Won, E.T., Baltzegar, D.A., Picha, M.E. and Borski, R.J. (2012) Cloning and characterization of leptin in striped bass (Morone saxatillis): Feeding inhibition and regulation by nutritional state. General and Comparative Endocrinology 178:98-107. 10.1016/j.ygcen.2012.04.019
- Sydorovych, O. and Daniels, H. (2011) Economic Analysis of Alternative Effluent Treatment Options for Pond Production of Hybrid Striped Bass in Aurora, North Carolina. Aquaculture Economics & Management. 15: 1, 46-70.
- Hiramatsu, N., Luo, W., Reading, B. J., Sullivan, C.V., Mizuta, H., Ryu, Y.-W., Nishiyama, O., Todo, T. and Hara, A. (2012) Multiple ovarian lipoprotein receptors in teleosts. Fish Physiol. Biochem. (in press) doi: 10.1007/s10695-012-9612-6
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Progress 06/15/10 to 06/14/11
Outputs
OUTPUTS: In the egg quality studies, the major system for uptake of yolk proteins by growing oocytes was characterized in detail in the striped bass. This system involves multiple yolk precursors, vitellogenins, that are produced by the liver and are taken up by oocytes after binding to specific receptors on the oocyte surface. Using combinations of sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting and mass spectrometry, the major yolk proteins derived from each of the 3 forms of vitellogenin (Vtg) were characterized and quantified in oocytes and ovulated eggs from females producing high quality eggs. The three forms of Vtg were also purified and used in competitive binding and ligand blotting experiments to develop evidence that the 3 Vtgs disparately bind to as many as three different receptors. The results of this research were reported to the scientific community and industry stakeholders in a special session on Aquaculture Genomics and in the Annual Meeting of the Striped Bass Growers Association at Aquaculture America 2011. The feasibility of determining the gender of Atlantic sturgeon via immunoassay of sex steroid hormones in their blood, urine, or mucus was assessed to provide a means by which male fish can be identified and culled early in the caviar production cycle. A local (NC) commercial sturgeon farming business collaborated in the research and production managers at the facility were given demonstrations of the use of plasma steroid ratios to determine gender in Atlantic sturgeon too immature to sex using ultrasound or minimally invasive biopsy methods. In studies conducted to evaluate how stress adversely affects the growth of farmed hybrid striped bass (HSB), radioreceptor assays were established to evaluate binding characteristics of the intracellular and putative membrane receptor for cortisol in fish. Additionally, extraction and radioimmunoassay procedures for measurement of cortisol in blood and tissue were validated. The results of this research were disseminated to the scientific community at the North American Society of Comparative Endocrinology. The feasibility of disposal of HSB farm pond effluents via land application was evaluated by using water from eighteen research ponds stocked with HSB to irrigate 24 plots of land planted with various tree species. Pond water was applied to the plots until standing water was observed and data on daily irrigation volumes and tree transpiration rates was collected year round. The results of this work have been presented to HSB producer groups at the annual N.C, Aquaculture Development Conference and in one-on-one meetings during numerous farm visits. PARTICIPANTS: Prof. Craig Sullivan, Department of Biology, NC State University (NCSU), Principal Investigator (PI); Prof. Harry Daniels, Department of Biology, NCSU, Co-PI; Prof. Jeff Hinshaw, Department of Biology, NCSU, Co-PI, Prof. Russell Borski, Department of Biology, NCSU, Co-PI; Prof. Tom Losordo, Department of Biological and Agricultural Engineering, NCSU, Co-PI; DR. Andy McGinty, Superintendent, NCSU Pamlico Aquaculture Field Laboratory; Dr. Benjamin Reading, Postdoctoral Research Associate, Dept. Biology, NCSU; Valerie Williams, Ph.D. graduate student, Dept. Biology, NCSU; Amanda Boury, Ph.D. graduate Student, Dept. Biology, NCSU; Justin Schilling, Ph.D. graduate Student, Dept. Biology, NCSU; Scott Salger, Ph.D. graduate Student, Dept. Biology, NCSU; Jamie Mankiewicz, M.S. graduate student, Dept. Biology, NCSU; Miller Johnstone, PhD graduate research assistant, Dept. Biology, NCSU; Brittany Holler, M.S. graduate student, Dept. Biology, NCSU; Christina Shenton, M.S. graduate student, Dept. Biology, NCSU; Katelyn Molloy, Undergraduate student, Dept. Biology, NCSU; Kathyrn Mills, Undergraduate student, Dept. Biology, NCSU; Melissa Pitachio, Undergraduate student, Dept. Biology, NCSU; Brad Ring, Research Specialist, Dept. Biology, NCSU; Brian Shannon, Research Specialist, Dept. Biology, NCSU; Michael Hopper, Research Specialist, NCSU Pamlico Aquaculture Field Laboratory; John Davis, Research Technician, Dept. Biology, NCSU; Adriane Gill, Research Technician, Dept. Biology, NCSU TARGET AUDIENCES: This research targeted reproductive physiologists, growth physiologists, endocrinologists, aquaculture scientists including engineers, aquaculture industry stakeholders, and government regulators of water resources and water quality. The project Principal Investigator and Co-Investigators all are also involved in classroom instruction in courses such as Fish Physiology, Aquaculture, and Endocrinology for which some of the research outlined here is used in formal lectures in the classroom. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
In the studies of egg quality in striped bass, the detailed characterization of precursor product relationships between the different forms of Vtg and yolk protein, and the discovery of multiple Vtg receptors, set the stage for evaluation of the degree to which these complex systems underlying yolk deposition are disrupted in females producing low quality eggs. A post-doctoral research scholar, a doctoral student, and an undergraduate student were trained in advanced proteomics methodologies for reproductive biology in this research. Measurement of sex steroids in pre-pubescent Atlantic sturgeon plasma is being used to develop ratios that discriminate between male and female sturgeon much earlier than gender can be identified using ultrasound, a current practice in the sturgeon industry. A commercial sturgeon producer collaborating with the project has been able to identify gender in groups of animals in which limited gonadal development precluded other methods of gender separation. In the studies of stress and growth physiology, assays for measuring the classical intracellular and a putative membrane glucocorticoid receptor abundance and affinity were established and utilized to show that cortisol may not only bind high affinity intracellular receptors to elicit its metabolic and osmoregulatory actions, but may rapidly modulate target cell activity through interactions with a novel membrane receptor. This receptor is pharmacologically distinct from the intracellular type and could be important for transducing rapid responses to the stress hormone, cortisol. Assays for measures of this key hormone indicative of stress responses in fish also were produced, allowing for assessment of environmental parameters that produce stress and that can subsequently impair growth potential, sex determination and osmotic responses of aquacultured fishes. These studies provided training on biochemical and endocrinological techniques for measuring levels of a stress steroid and its tissue sensitivity in fish to three graduate and two undergraduate students. North Carolina State regulators in the Division of Water Quality are monitoring the results of the research on land application of HSB pond effluents and the final results will be used to help establish Best Management Practices for HSB pond culture in North Carolina; this land-application is a novel approach to the treatment of aquaculture effluents. The establishment of this research tree plot provides us with a unique capability to evaluate these systems for the removal of environmental contaminants from fish production.
Publications
- Johnstone W, Alyea R, Mills K, Thomas P and Borski R (2011). Identification of a putative membrane glucocorticoid receptor in the Mozambique Tilapia (Oreochromis mossambicus. Front. Endocrin. Conference Abstract: NASCE 2011: The inaugural meeting of the North American Society for Comparative Endocrinology. doi:10.3389/conf.fendo.2011.04.00147
- Reading, B.J., Hiramatsu, N., and Sullivan, C.V. 2011. Disparate Binding of Three Types of Vitellogenin to Multiple Forms of Vitellogenin Receptor in White Perch. Biology of Reproduction 84(2):392-399.
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