Duration: 10/01/2007 to 09/30/2013

Administrative Advisor(s):

NIFA Reps:

Non-Technical Summary

Statement of Issues and Justification

The channel catfish, Ictalurus punctatus, is the primary cultured fish for food production in the US representing approximately 70% of all US aquaculture production. Catfish production has stagnated at about 300 million kg annually during the last 5 years in an industry that was once doubling in size every 10 years. The industry is currently struggling to keep pace with the increasing cost of inputs and competition with inexpensive imported fish. The catfish industry includes both large and small family farms. In many impoverished areas of the South, the catfish industry is critical for the economic viability of the rural communities. In these areas, the catfish industry both directly and indirectly provides one of the few opportunities for communities, rural minorities and women via all aspects within the industry and supporting services. Catfish production generates some $500-million in farm revenue and backward and forward linkages raise the value of the industry to an estimated $4-billion. Stakeholders indicate that catfish farming needs to be more productive, efficient, competitive, sustainable and profitable, and some have gone out of business. Given the trade deficit for edible fish products in the US reached an all time high of $10.7 billion in 2000, domestic production of fish is critical to reduce trade deficits and maintain food security. Stakeholders indicate that lowering absolute production cost to a level at which post-production transportation and marketing costs become relatively more significant than labor is key for revitalization of the American catfish farm and to overcome pressure form foreign imports. A high impact solution is the utilization of hybrid catfish, which is desired by the stakeholders. However, stakeholders need optimized hybrid embryo production technology, increasingly improved hybrids and specific hybrid harvesting technology. Significant progress has been made in improving the technology for producing hybrid catfish embryos. Despite these significant improvements, hybrid embryo production technology needs further improvement to optimize hybrid fry production and allow large-scale adoption of the hybrid in an industry needing about 1 billion fry per year. Depending upon conditions, hybrid food fish production has the potential for $700- $2,800 more profit/acre compared to channel catfish food fish production. The consequences of not conducting this research and solving this problem are a slowing of the adoption of the hybrid technology, significant economic loss by all sectors of the catfish industry, increasing loss of revenue to foreign imports and threats to the sustainability and survival of the American catfish industry. Some stakeholders have stated that  the hybrid is the only hope for the survival of the American catfish industry.

Social impact is great as it will help preserve rural communities and jobs for minorities and women in the South. The outcome should enhance global competitiveness which is critical concerning our current situation with fish product imports and the influx of catfish and fish from other countries. The benefit to the industry could easily be one billion $ per year if the hybrid could immediately be applied across the entire industry.

The advantage of using the multistate approach is two-fold; this will allow a larger effort, allowing more rapid progress and implementation, and different expertise exists in the proposed participating states, increasing effectiveness, output and enhancement of the multi-disciplinary approach.

Related, Current and Previous Work

The hybrid resulting from the mating of female channel catfish, Ictalurus punctatus, X male blue catfish, Ictlaurus furcatus, is the best catfish for pond culture. The potential growth advantage of the channel catfish X blue catfish hybrid in the pond environment was first noted by Giudice (1966); however, stocking rates used were well below that utilized in commercial catfish culture. Yant et al. (1975) first demonstrated the increased growth rates of hybrids compared to channel catfish at commercial densities in earthen ponds. Chappell (1979) and Dunham et al. (1990) subsequently demonstrated a similar superiority of the hybrid compared to blue catfish under commercial conditions. Feed conversion of the hybrids averaged 1.35:1 while feed conversion of channel catfish averaged 1.56:1(Yant et al. 1975). Since these earlier experiments, it has been demonstrated that hybrid catfish always exhibits heterotic growth in ponds compared to its parental species (Dunham et al. 1987, 1990; Jeppsen 1995; Dunham and Brummett 1999). Growth improvement of hybrids over that of channel catfish ranged from 20-100% and is more pronounced with high stocking densities. Increased growth of hybrids has been demonstrated at both the fingerling and food fish stages. As density increased, growth advantages of the hybrid catfish emerged compared to channel catfish and the percent difference in size increased with increasing density. Yant (1975) and Brooks et al. (1982a, b) found more uniform body weight of catfish hybrids compared with channel catfish, and Dunham et al. (1982) demonstrated the body shape of hybrids was more uniform than that of channel catfish. Carcass traits such as dress out percentage and fillet percentage of hybrid catfish are also superior to channel catfish (Yant 1975; Chatakondi et al. 2000).).
Disease resistance and survival of hybrid catfish is clearly superior at all life stages (Ella 1984; Jeppsen 1995; Dunham and Brummett 1999). High stocking densities often result in poor water quality and disease outbreaks that adversely affect the survival of channel catfish more than hybrid catfish (Dunham et al. 1990). Ella (1984) demonstrated that the hybrid was more resistant to columnaris, Flavobacterium coumnare, and aeromonas, Aeromonas hydrophila than channel catfish. Dunham (unpublished) demonstrated that hybrids had higher survival when infected with ich, Ichthyoptheius mulitfilis, than channel catfish. Wolters et al. (1996) determined that survival after being challenged with an emersion bath of Edwardsiella ictaluri was 89.5% for blue catfish, 73.8% for hybrid catfish, and 62.0% for channel catfish. Injections of E. ictaluri were lethal only to channel catfish (Wolters et al. 1996). Antibody responses for bath and injection challenges of E. ictaluri were lowest in blue catfish, intermediate in hybrid catfish, and highest in channel catfish.
The hybrid also demonstrated increased tolerance to low dissolved oxygen conditions. When exposed to low dissolved oxygen in concrete tanks, 100% mortality of channel catfish was observed, compared to 33% mortality of hybrid catfish (Dunham et al. 1983). Additionally, the hybrid appears to be able to find aerators more easily than channel catfish when low oxygen events occur. The hybrid is easier to harvest by seining, corral seine trapping, and angling by hook-and-line than channel catfish. A single seine haul captured 75% of hybrid catfish; whereas at least two seine hauls were necessary to capture 75% of channel catfish (Yant 1975; Chappell 1979; Dunham and Argue 1998). Tave et al. (1981) showed that hybrid catfish were more easily caught by hook-and-line angling than blue catfish, channel catfish, or the reciprocal hybrid.
Large-scale field trials have confirmed the outstanding performance of the hybrid in commercial settings and in processing plants, resulting in very high demand and premium prices for fingerling hybrids. The strain of parent affects the performance of the hybrid. In general, better performing strains of channel catfish result in better performing genotypes of hybrids (Dunham et al. 1987; Ramboux 1990).

Behavioral reproductive isolating mechanisms between blue catfish and channel catfish have prevented consistent commercial scale production of hybrid catfish, however there are no gametic blocks for making this hybrid. The fertilization rate and the hatching rate of the channel x blue hybrid embryos and the fry production per kg of female are similar to the channel catfish pairs when the two genotypes are produced using the same technique (Bart 1994; Dunham et al. 1999; Lambert et al. 2000).Artificial hand stripping and fertilization technologies have been developed that allow small-scale production of the channel-blue hybrid (Bart 1994, Dunham et al. 1999; Lambert et al. 2000). However, today only a few farmers utilize the hybrid because fingerlings are not available and because of the difficulty in making hybrid eggs and fry.

During the last 28 years Rex Dunham has been leading research for developing the basic technology platform that we are trying to improve in this proposal. Dr. Anita Kelly, Southern Illinois University has been examining they effect of estrogenic like compounds in catfish diets on the sexual development of blue catfish males used to make hybrids. Dunham currently leads a multistate USDA SRAC project on hybrid embryo production. Other participants are Dr. Nagaraj Chatakondi, Eagle Aquaculture, Allen Davis and Ron Phelps, Auburn University, Terrence Tiersch, Louisiana State University, Lou Dabramo, Mississippi State University, Brian Small and Brian Bosworth, CGRU USDA, Stoneville, Bill Simco and Charles Lessman, University of Memphis researching various aspects of hybrid embryo production. This project will end in 1.5 years. The proposed multistate project will build upon the accomplishments of the SRAC project, and serve as a mechanism to continue the multistate effort on improved hybrid embryo production and performance.

Highlights of SRAC progress follow: Early spawning can be accomplished by heating water prior to the natural spawning season without any difference in success compared to the natural spawning season. When 100 degree hours are reached ovulation and fertilization should be successful. If warm water is available, channel catfish can be successfully spawned in early January. Feeding standard 32% protein floating catfish feed 6 times per week for 2 months prior to spawning gives equal or better fry production compared to high protein diets. Supplementation of brood fish diets with menhaden fish oil, DHA and ARA two months prior to spawning can increase hybrid fry output 33-100% depending upon previous preparation of the fish. Availability of at least low levels of forage fish can have some positive impact on hybrid fry production. Utilization of the appropriate genetic line of channel catfish female can double and triple hybrid fry output. Strain of blue catfish male impacts hatch rate of hybrid embryos and sperm production. The utilization of 100 Fg/kg LHRHa implants to induce channel catfish females most consistently generates the greatest number of fry/kg. The dosage of both implants and injections needs to be reduced as the season progresses to maximize fry production. Indirect exposure of females to water containing conspecifics males increases fry production. Hatch rate of hybrid embryos is improved if channel catfish females are stripped within 2 hours of first observation of egg release. Ultrasound may be used to ascertain ovulation in channel catfish females and the appropriate time for stripping of eggs. Spectrophotometric assays can be used to determine sperm concentrations from crushed testis of catfish. Utilization of this tool should result in more efficient use of sperm, and more consistent fertilization rates. Sperm concentrations can be reduced 100-fold, from 2 X 10 8 to 2 X 10 6 sperm/800 eggs, and still obtain good hatch rates. The cost for the sperm savings is a reduction of 16% for relative hatch rate. The frequency of formalin treatments should be three times per day to maximize hatch rate of hybrid embryos and four treatments per day is excessive. At 28EC, hybrid embryos are chemically sensitive to formalin between 42 to 46 hours post-fertilization, and formalin treatments should be avoided during this period to maximize hatch rate.

In the proposed multistate project objectives 1,7,8 and 9 were examined in the SRAC project.The progress was good in each of these areas, however, there is potential for further study and greater advancement from expanding on these earlier studies. The remaining 9 objectives were not in the SRAC project,but allow a more holistic, and a greater multidisciplinary approach to the problem compared to the SRAC project. The multistate will also be broader in nature examining further improvement of the hybrid, developing easier harvest methods for the hybrid, reducing hormone costs for the hybrid as well as the reproductive aspects of making fry. The SRAC project focused solely on the reproductive aspects of making the hybrid fry.

Duplication- A CRIS search was conducted and no duplication was found.

CRIS was searched using the key words hybrid catfish and 13 hits were found. The search indicated that the proposed project is unique. The other projects found were our own research, joint collaboration with others, expired projects or on hybrid research that differs from ours including gene mapping, evaluation of gene maps, ecological studies or other culture evaluations.

1. ACCESSION NO: 0409121 SUBFILE: CRIS ,INVESTIGATOR: Liu, Z. IDENTIFICATION OF DNA MARKERS FOR GENOTYPING SUB-SPECIES POPULATIONS OF FISH

2 ACCESSION NO: 0182696 SUBFILE: CRIS, INVESTIGATOR: Dunham, R. A.; Liu, Z. GENETIC IMPROVEMENT OF CATFISH

3. ACCESSION NO: 0193714 SUBFILE: CRIS, INVESTIGATOR: Liu, J.

4 ACCESSION NO: 0196602 SUBFILE: CRIS, INVESTIGATOR: Daniels, W. H.; Chappel, J. EVALUATION OF AQUATIC SPECIES, STRAINS AND HYBRIDS AND THEIR PRODUCTION METHODS TO IMPROVE SUSTAINABILITY OF AQUACULTURE IN ALABAMA

5. ACCESSION NO: 0198024 SUBFILE: CRIS, INVESTIGATOR: Davis, D. A. NUTRITION AND FEED MANAGEMENT FOR WARM WATER FISH AND SHRIMP.

6. ACCESSION NO: 0209590 SUBFILE: CRIS ,INVESTIGATOR: Phelps, R. P. TECHNIQUES TO IMPROVE NATURAL HYBRIDIZATION OF CHANNEL CATFISH FEMALES WITH MALE BLUE CATFISH

7. ACCESSION NO: 0186397 SUBFILE: CRIS, INVESTIGATOR: Burtle, G. J.; Lewis, G. W. HYBRID CATFISH TECHNOLOGY DEMONSTRATION AND EXTENSION FOR FARM SUSTAINABILITY

8 ACCESSION NO: 0182266 SUBFILE: CRIS, INVESTIGATOR: Morrison, D. G. PRODUCTION AND ECONOMIC ENHANCEMENT OF LOUISIANA'S AQUACULTURE INDUSTRY

9 ACCESSION NO: 0189295 SUBFILE: CRIS, INVESTIGATOR: Tiersch, T. R.
DEVELOPMENT, MAINTENANCE, AND DISTRIBUTION OF GENETIC IMPROVEMENT IN AQUACULTURE
10. ACCESSION NO: 0404545 SUBFILE: CRIS, INVESTIGATOR: TORRANS E L; ZIMBA P V OPTIMIZING CATFISH/WATER QUALITY INTERACTIONS TO INCREASE CATFISH PRODUCTION EFFICIENCY

11 ACCESSION NO: 0402038 SUBFILE: CRIS, INVESTIGATOR: WOLTERS W R; WALDBIESER G C; BOSWORTH B G; BILODEAU A L; SMALL B C; VACANT CATFISH GENETICS, BREEDING, AND PHYSIOLOGY

12 ACCESSION NO: 0207416 SUBFILE: CRIS, INVESTIGATOR: Hanson, T. R. ECONOMICS OF AQUACULTURE PRODUCTION IN MISSISSIPPI

13 ACCESSION NO: 0192175 SUBFILE: CRIS, INVESTIGATOR: Yant, R. SPAWNING AND HATCHERY TECHNOLOGY TO IMPROVE HYBRID CATFISH FRY PRODUCTION

Objectives

1. Improve hybrid embryo production by determining the best nutritional regime to maximize fecundity and hatch rate from induced channel catfish females and blue catfish males.
   
2. Conduct family selection for fecundity, hatch rate of hybrid embryos and hybrid fry produced/kg for channel catfish females that were hormone induced and hand stripped.
   
3. Lengthen the short spawning season for hybrids by developing and identifying early, peak and late spawning lines of channel catfish.
   
4. Determine the relative importance of stocking density of channel catfish females for hybrid embryo production- should density be based on surface acreage or volume?
   
5. Compare various forms of LHRH implants to address INAD concerns and reduce implant costs.
   
6. Determine the ideal temperature for ovulation hybrid embryo hatching in best late, peak and early spawning strains.
   
7. Improve hybrid embryo production via pheromonal manipulation of channel catfish males and blue catfish males for improved ovulation, spermiation, egg quality, hatch and fry production .
   
8. Develop extended refrigerated storage and cryopreservation of sperm.
   
9. Evaluate morphological changes of oocytes during oocyte maturation in female catfish and relate this to fertilization, development and hatch.
   
10. Conduct recurrent family selection for growth, feed conversion, disease resistance, oxygen tolerance , harvestability and carcass traits of hybrid catfish.
    
11. Develop in pond seining/grading technology specific for hybrid catfish to prevent gilling
    
12. Develop technology to ejaculate blue catfish.
    
13. Conduct gene transfer to improve growth, feed conversion, disease resistance and body composition and transgenic sterilization of hybrid catfish.
    

Methods

Several institutions are addressing various objectives. Different institutions have expertise in specific areas, and thus will lead specific efforts. In this manner, a coordinated framework will be constructed of a multi-disciplinary approach to improve the key aspects of the overall technology of hybrid embryo production and implementation of hybrid technology. In some cases, multiple institutions will conduct parallel experiments and in some cases, different steps in an experiment will be conducted at different institutions. Minimally,a yearly planning meeting will be conducted by the technical committee and a multi-state annual report will also be issued. Objective 1) ) Improve hybrid embryo production by determining the best nutritional regime to maximize fecundity and hatch rate from induced channel catfish females and blue catfish males. These will be follow-up studies based on preliminary work conducted at Southern Illinois University and Auburn University. Fatty acid enhanced diets, soybean less diets and forage will be evaluated in ponds to maximize fecundity and hatch rate from induced channel catfish females and blue catfish males.36% protein feed will be used because unpublished results of Dr. Chatakondi indicate that this is superior to 28% protein. Four treatments will be evaluated in the first experiment: a) a standard 36% protein catfish feed, b) 36% protein catfish diet without soybeans, c) forage fish only (1000kg/ha standing crop of tilapia will be established) d) 36% protein catfish diet without soybeans with supplemental forage fish and a fatty acid enhanced 36% protein diet. Proximate analysis of tilapia will be conducted to determine their nutrient quality. No research has addressed comparing summer, spring or year-round enhanced nutrition for fry output and the relative cost/benefit of using these more expensive diets during these different time periods. In the second experiment, fatty acid enhanced diets (FA) (36% protein) will be compared to control (36% protein) in all possible seasonal combinations for summer feeding(June-January) and spring feeding (February-May).Four treatments will be compared. Season Diet regime Summer Spring Control #1 36% 36% #2 FA 36% #3 36% FA #4 FA FA Ten replicate fish will be stocked at 1,000kg/ha in 3 replicate ponds for a total of 30 replicate fish per treatment. Fish will be fed ad libitum five times per week. Artificial fertilization procedures to produce hybrid embryos will be conducted. Parameters measured will be testes weight, gonadosomatic index, sperm/g testes, sperm/kg body weight, ovulation rate, fecundity, hatching rate and fry/kg. Dr. Davis will design the diet formulations and prepare brood stock at Auburn University. Dr. Kelly will conduct a parallel subset of experiments at Southern Illinois University with various aged fish focusing primarily on males Drs. Dunham, Kelly and Chappell will conduct the spawning research Planning and implementation (Auburn University, Southern Illinois University). Objective 2) Conduct family selection for fecundity, hatch rate of hybrid embryos and hybrid fry produced/kg for channel catfish females that were hormone induced and hand stripped. This objective will be addressed at Auburn University. Every channel catfish male and female in the evaluaton will be mated to two individuals of the same species generating 200 full sib- half-sib channel catfish families to perform selection upon. Additionally, every channel catfish female is mated to two blue catfish males creating 200 full sib-half sib families to measure response to selection for reproductive traits in their channel catfish dam. Representatives of each channel catfish family are then raised to maturity and mated to blue catfish in to generate the data set needed to evaluate recurrent selection and family selection. A randomly bred channel catfish control line from the original base population will also be perpetuated and the females mated with blue catfish males to provide the control for selection response of the reproductive traits. Individual hybrid egg masses will be incubated separately utilizing standard procedures in paddlewheel troughs. One day prior to hatch, the hybrid egg masses will be weighed, mean egg weight and % alive embryos determined to estimated hatch. Parameters measured will be percent of females gravid, percent of females ovulating, relative fecundity, fertilization rate at 48 hrs, hatching rate, and fry produced/kg female. Realized heritability, h2 = R/S; where R= response to selection and S= selection differential: will be calculated for fecundity, hatch and fry/kg. Standard error of the response to selection, ÃR, will be calculated as the square root of (Vp[(h2/Ne)+(1/m)]R) / S where Vp, = phenotypic variance, Ne = the effective population number, and m = the number of individuals measured (Falconer and Mackey 1994). Objective 3) Lengthen the short spawning season for hybrids by developing and identifying early, peak and late spawning lines of channel catfish. University of Minnesota will coordinate brood stock collection and Auburn University will conduct the spawning experiments (Auburn University, University of Minnesota). The 3 treatments will be Minnesota, RioGrande and AU-3 strains of channel catfish females. Females from these 3 strains will be hybridized with blue catfish males.Twenty females per strain will be implanted with LHRHa for ovulation on April 15, representing the early spawning season. The experiment will be repeated on May 21 and July 1 representing peak and late spawning season. Parameters measured will be percent of females ovulating, relative fecundity, hatching rate, and hybrid fry produced/kg female. Objective 4) Determine the relative importance of stocking density of channel catfish females for hybrid embryo production- should density be based on surface acreage or volume?(Eagle Aquaculture, Mississippi State University, Auburn University). Four treatments will be evaluated both stocked at 1,500 kg/ha with 4 replicates and all ponds average about 3 surface ha. Treatment 1 - Channel catfish females prepared in deep hill ponds at 1,500/kg. Treatment 2- channel catfish females prepared in levee ponds at 1,500kg/ha. Treatment 3 - Channel catfish females prepared in deep hill ponds at 750/kg/m and Treatment 4-channel catfish females prepared in levee ponds at 750 kg/ha/m.Experiments will be conducted by Dr. Chatakondi (Eagle Aquaculture and affiliate professor at MSU and AU) and Dr. Dunham at both Eagle Aquaculture and Auburn University. Objective 5) Compare various forms of LHRH implants to address INAD concerns and reduce implant costs. (University of Maryland, Auburn University). Six treatments will be evaluated. Channel catfish females (20 replicate females per treatment) will be implanted with 100ug/kg of a) fresh LHRHa, b) LHRHa after one year of storage, c) extruded LHRHa implants, d) reconstituted LHRHa implants, e) cellulose implants or f) Ovaplant. The Department of Polymer and Fiber Engineering, Auburn University, will develop protocols and execute the extrusion of LHRH implant (EVAC implants from Univ. of Maryland) and extrusion of cellulose based implants which may be more amenable to FDA approval. University of Maryland will provide fresh and reconstituted implants. Ovaplant is a Syndel product currently being evaluated for FDA approval. Parameters measured will be percent of females ovulating, relative fecundity, 48 hour ferilization, hatching rate, and hybrid fry produced/kg female. AU will conduct the spawning experiments. Objective 6) Determine the ideal temperature for ovulation hybrid embryo hatching in best late, peak and early spawning strains. Currently, it is recommended that induced spawning for hybrid embryo production be conducted at 28C. However, this data was generated on a very poor performing strain for artificial production of hybrid embryos. Ideal temperature for artificial production could vary among strains adapted for spawning at different temperatures. Late, peak and early spawning channel catfish females from year 1 will be induced for hybrid production at 24, 26, 28 and 30C to determine ideal spawning temperatures for catfish of varying reproductive characteristics. Hatching temperatures will correspond to spawning temperatures.If differences exist, this will impact both management strategies as well as energy costs. Complementary experiments will be conducted at LSU, Eagle Aquaculture and AU. Objective 7) Improve hybrid embryo production via pheromonal manipulation of channel catfish males and blue catfish males for improved ovulation, spermiation, egg quality, hatch and fry production (University of Memphis, Auburn University,USDA). University of Memphis and USDA will focus on the isolation of potential pheromonal compounds from testes and ovaries utilizing HPLC or GC. Candidate compounds will be compared to holding water containing conspecifics of the opposite sex and unexposed holding water for their effects on ovulation, sperm production and hatching rate. Six replicate holding tanks will be used per treatment Objective 8) Develop extended refrigerated storage and cryopreservation of sperm (Louisiana State University). Refrigerated storage - Functional methods exist for refrigerated storage of blue catfish sperm, but these methods require optimization. We will focus on simple extender formulations but will use one week of storage as our target baseline. Additionally, standardized procedures for aseptic collection of testis and preparation of sterile extender solutions will be developed. The efficacy of antibiotics in fresh storage of sperm will be evaluated. Hatch rate and fry per kg of female body weight will be determined for comparison of refrigerated storage and fresh sperm treatments. Determination of sperm-to-egg ratios will be delineated. Cryopreservation - The sperm samples will be evaluated by microscopic examination, and high quality samples will be cryopreserved using established techniques at the LSU Dairy Improvement Center. Procedures will be developed to streamline this process and to maximize efficiency and improve production suitable for commercial application. Based on our previous work, we anticipate focusing our efforts on evaluation of 5% and 10% concentrations of methanol and dimethyl sulfoxide. Ferilization estimate at 48 hr,hatch rate and fry per kg of female body weight will be determined for comparison of cryopreservation and fresh sperm treatments. Objective 9) Evaluate morphological changes of oocytes during oocyte maturation in female catfish and relate this to fertilization, development and hatch. Morphological changes of oocytes during oocyte maturation in female catfish and relate this to fertilization, development and hatch will be studied using computerized scanning (University of Memphis, LSU, Auburn University). Scanning procedures, Computer-Aided Screening (CAS) and Computer-Aided Meiotic Maturation Assay (CAMMA), will be used to follow development of embryo arrays over time. AU and LSU will produce embryos. University of Memphis will conduct the scanning and analysis of the images. The CAS technique will be applied to catfish eggs that have been stripped from treated females and fertilized in vitro. Clutches of fertilized eggs from different females having different hormonal treatments will be arrayed on grid-lined dishes which spatially separate the clutches from one another and allow imaging at preset intervals throughout the developmental period of several days. Subsequent image analysis of the stacked images will allow the quantification of normal, abnormal and dead embryos over time. Thus, data will be obtained determining the quality of the original zygotes, which can then be related back to the treatments used. Objective 10) Conduct recurrent family selection for growth, feed conversion, disease resistance, oxygen tolerance , harvestability and carcass traits of hybrid catfish (Auburn University). The same fish spawned for objective 2 are spawned and used for this objective , thus 1,100 hybrid families are generated. From this pool representing all 100 original females, the select hybrid progeny for each trait will come from the top 10% of the select F2 channel catfish females (10 females (5 from each of the 2 half-sib families) X top 10 performing females= 100 select hybrid progenies compared to 1,000 control hybrid progenies) for growth, feed conversion, disease resistance, oxygen tolerance , harvestability and carcass traits of hybrid catfish. Data generated for each trait allows calculation of response to individual selection (realized h2) for combining ability of channel catfish females. The data generated also allows comparison of family and individual selection. Data from each trait from each individual hybrid family will be classified and used as data for the select population or the control population independently depending upon the performance of the hybrid progeny from their grand dam. Objective 11) Develop in pond seining/grading technology specific for hybrid catfish to prevent gilling (University of Arkansas Pine-Bluff). UAPB has developed an in pond grader for catfish requiring pumping. A second system will be evaluated by modifying seines and grading socks without a pumping system. These 2 systems will be compared for the efficacy in grading hybrid catfish without gilling them and in preventing mortality of fish passing through the grader.Six replicat trials will be conducted per grader system. Objective 12) Develop technology to ejaculate blue catfish. (University of Minnesota, Southern Illinois University and Auburn University). Testes structure will be studied and electrical and neuropeptide technologies evaluated to induce ejaculation in blue catfish. SIU will evaluate the electroejaculation. UM will evaluate a variety of neurotransmittors for their effect on contraction and relaxation of testes and possible electroejaculation in vitro and in vivo. AU will provide testes and fish for UM. Objective 13) Conduct gene transfer to improve growth, feed conversion, disease resistance and body composition and transgenic sterilization of hybrid catfish. (AU) This complements and enhances existing objective 10 and solidifies the elimination of environmental issues as well as possibly leading to a specialized, high value, healthy heart hybrid catfish and better control of reproduction. Hybrid catfish will be produced contain growth hormone, cecropin, desaturase and sterilizations constructs. The growth, feed conversion, disease resistance, body composition and reproductive ability of these fish will be compared to non-transgenic controls in replicated government approved confinement ponds.

Measurement of Progress and Results

Outputs

• A stable, sustainable, cost effective hybrid embryo technology.
• Genetically improved lines of channel catfish and blue catfish for hybrid production.
• A longer spawning season for producing hybrid embryos.
• A better understanding of the process of synchronizing the ovulation of high quality ova.
• A grader/seine specifically for efficient hybrid harvest.
• Output 6 Data sets and publications on hybrid catfish. Output 7 Wide spread adoption of hybrid catfish technology.

Outcomes or Projected Impacts

• Social impact is great as it will help preserve rural communities and jobs for minorities and women in the South.
• The outcome should enhance global competitiveness which is critical concerning our current situation with fish product imports and the influx of catfish and fish from other countries.
• The benefit to the industry could easily be one billion $ per year if the hybrid could immediately be applied across the entire industry
• Catfish farming should become even more environmentally friendly through reduced use of chemicals, antibiotics and less possibility of interbreeding with wild stocks.
• A healthier product will result for consumers.

Milestones

(2007): 1) Collect brood stock in 2007 to identify early, peak and late spawning lines of channel catfish in 2007. 2)Develop extrusion process for LHRHa implants and determine release rates in 2007 to allow evaluation of implants in 2007.3)Determine response to selection and realized h2, for fecundity, hatch and hybrid fry/kg based on individual selection in 2007 to determine selection protocol for 2008,4)Compare various forms of LHRH implants to address INAD concerns and reduce implant costs in 2007 to have final spawning protocol in 2011, 5)Develop extended refrigerated storage and cryopreservation of sperm in 2007 to have final spawning protocol in 2011,6) Develop and identify early, peak and late spawning lines of channel catfish in 2007 to lengthen the short spawning season for hybrids by 2011 7) Evaluate morphological changes of oocytes during oocyte maturation in female catfish and relate this to fertilization, development and hatch in 2007 to determine key developmental stages for study in 2008. 8) Evaluate current pond graders for effectiveness for hybrids in 2007 to determine design changes for 2008.

(2008): 9) Construct altered grader in 2008 to evaluate pond seining/grading technology specific for hybrid catfish to prevent gilling in 2009. 10)Enhance fatty acids diets evaluated by 2008 to have final spawning protocol in 2011. 11) Determine the relative importance of stocking density of channel catfish females for hybrid embryo production- should density be based on surface acreage or volume?: in 2008 to have final spawning protocol in 2011. 12) Evaluate hybrid embryo production via pheromonal manipulation of channel catfish males and blue catfish males for improved ovulation, spermiation, egg quality, hatch and fry production in 2008 to have final spawning protocol in 2011

(2009): 13) Determine the ideal temperature for ovulation hybrid embryo hatching in best late, peak and early spawning strains in 2009 to have final spawning protocol in 2011. 14)Forage supplements evaluated by 2009 to have final spawning protocol in 2011. 15) Evaluate family selection for fecundity, hatch rate of hybrid embryos and hybrid fry produced/kg for channel catfish females that were hormone induced and hand stripped in 2009 to have improved brood stock in 2011.16) Have key growth, disease resistance, body composition and sterilization genes transferred to channel and blue catfish in 2009 to allow transgenic hybrid evaluation in 2012

(2010): 17) Non-soybean diets evaluated by 2010 to have final spawning protocol in 2011. 18) Evaluate recurrent family selection for growth, feed conversion, disease resistance, oxygen tolerance, harvestability and carcass traits of hybrid catfish 2010 to allow multiplication of improved brood stock in 2011.

Projected Participation

View Appendix E: Participation

Outreach Plan

Results of this project will be made available via presentations at catfish farmers meetings, demonstrations/workshops, extension publications, dissemination of protocols and peer reviewed publications.

The project will produce an annual scientific report. The report will have a technical section as well as an extension/recommendations section. This report will be posted on a web site developed for the project and distributed to state and national catfish farming organizations as well as to southern aquaculture extenison agents/ programs. Participants will be encouraged to contribute to the research forums of the annual US World Aquaculture Meeting and the Catfish Farmers of America Research Forum. At the projects end a cumulative final report will be made and distributed in the same manner.

Organization/Governance

The recommended Standard Governance for multistate research activities include the election of a Chair, a Chair-elect, and a Secretary. All officers are to be elected for at least two-year terms to provide continuity. Administrative guidance will be provided by an assigned Administrative Advisor and a CSREES Representative.

The project will have an annual meeting to allow the participants/collaborators to report and discuss progress,solidify collaborative relationships and research, discuss how to overcome pitfalls and plan the upcoming years research.

Literature Cited

Bart, A. N. 1994. Effects of sperm concentration, egg number, fertilization method, nutrition and cryopreservation of sperm on fertilization efficiency with channel catfish eggs and blue catfish sperm. Ph.D. Dissertation, Auburn University, AL, USA.

Brooks, M. J., R. O. Smitherman, J. A. Chappell, and R. A. Dunham. 1982a. Sex-weight relations in blue, channel, and white catfishes: implications for brood stock selection. Prog. Fish-Cult. 44:105-106.

Brooks, M. J., R. O. Smitherman, J. A. Chappell, J. C. Williams, and R. A. Dunham. 1982b.Length variation in species and hybrid populations of blue, channel and white catfishes. S.E.Assoc. Fish and Game Commis. 36:190-195.

Chappell, J. A. 1979. An evaluation of twelve genetic groups of catfish for suitability in commercial production. Ph.D. dissertation, Auburn Univ., AL.

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Attachments

Land Grant Participating States/Institutions

AL, AR, MD, MN, MS

Non Land Grant Participating States/Institutions

Eagle Aquaculture, Southern Illinois University, University of Memphis, USDA-ARS