REFINING CULTURE METHODS FOR SELECT MARINE BAITFISH

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: HATCH
• Reporting Frequency: Annual
• Accession No.: 0228674
• Project Start Date: Apr 1, 2012
• Project End Date: Sep 30, 2017
• Program Code: [(N/A)]- (N/A)

Project Director
Ohs, CO.

Recipient Organization
UNIVERSITY OF FLORIDA
G022 MCCARTY HALL
GAINESVILLE,FL 32611

Performing Department
Indian River Research and Education Center

Non Technical Summary
Global fisheries yields approximately 80 million metric tonnes of food fish each year and this amount has not varied greatly for 30 years. On the other hand, aquaculture of food fish continues to experience growth, and in 2010 accounted for nearly 51% of all food fish produced. Aquaculture can be a sustainable option to provide the world with protein and other products while alleviating pressure on wild fisheries. Aquaculture production in North America increased over 3.0% annually from 1995 through 2005. Baitfish aquaculture is a significant industry in the U.S. because it serves the 40 million anglers who have an estimated impact of nearly $125 billion on the nation's economy. Over 25 million freshwater anglers in the U.S. generated around $31 billion in revenue in 2006, with an estimated contribution of $87 billion to the nation's economy. There was over $38 million in sales of cultured freshwater baitfish in 2005, largely composed of the fathead minnow (Pimephales promelas), and golden shiner (Notemigonus crysoleucas).. Successful markets for these and other species is a direct result of the popularity of freshwater fishing throughout the U.S. Although marine baitfish aquaculture is in its infancy compared to freshwater baitfish, a strong market potential exists. In 2006, there were over 8.5 million saltwater anglers in the U.S. who generated $11 billion in sales resulting in an estimated $30 billion contribution to the economy. The Southeastern U.S. states from North Carolina to Texas had over 5 million licensed saltwater anglers in 2006, resulting in a $10 billion contribution to the economy. Florida had two million licensed saltwater anglers in 2006, the most of any state, with an estimated impact of $5.1 billion to the nation's economy. In 2005, the number of fish farms and the value of aquacultured products in Florida represented only 8.3% and 5.3%, respectively, of the U.S. total. Florida's largest aquaculture industry is the production of ornamental fish. However, strong potential exists to develop a new, high value marine baitfish aquaculture industry. Freshwater baitfish generally command a higher price per pound than other fish species, even when compared to ornamentals. A survey conducted in Florida indicated periods of high demand for certain sizes and species of live, marine baitfish throughout the year. Surveys in other states have shown similar trends. Culture of these high demand marine baitfish could support a growing aquaculture industry and reduce pressure on natural populations while providing a consistent supply of live, health baitfish to anglers. In 2004, a baitfish workshop held in Ruskin, Florida identified mudminnows (Fundulus spp.), spot (Leiostomus xanthurus), pinfish (Lagodon rhomboides), pigfish (Orthopristis chrysoptera), and croaker (Micropogonias undulatus) as having strong potential for successful culture. Extension publications on production of several species are available, but for most species, additional research is needed to elucidate larval nutrition, larval culture, and spawning biology.

Animal Health Component

(N/A)

Research Effort Categories

Basic

(N/A)

Applied

(N/A)

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
301	3799	1010	34%
302	3799	1010	33%
511	3799	1020	33%

Knowledge Area
302 - Nutrient Utilization in Animals; 511 - New and Improved Non-Food Products and Processes; 301 - Reproductive Performance of Animals;

Subject Of Investigation
3799 - Cultured aquatic animals, general/other;

Field Of Science
1010 - Nutrition and metabolism; 1020 - Physiology;

Keywords

aquaculture

marine baitfish

spawning

larval culture

pinfish

pigfish

minnow

goggle eye

copepod

lagodon

orthopristis

fundulus

Goals / Objectives
I) Develop spawning protocols for pinfish, pigfish, goggle eye, ballyhoo, and Seminole killifish. Research photo-thermally controlled cycles, identify optimal hormone agents and their respective dose rates for pinfish, pigfish and goggle eye, and determine spawning substrate preference for ballyhoo; II) Develop larval culture protocols for pinfish, pigfish, goggle eye, and Seminole killifish by researching responses to environmental conditions, specific dietary nutrient levels, feeding live prey items and artificial diets, and establishing optimal stocking densities for growth and survival of larvae; III) Develop low salinity culture protocols by identifying required mineral levels for juvenile pinfish and pigfish that promotes grown to market size in low salinity water; IV) Determine the market potential of cultured marine baitfish in the state of Florida through bait retailer and angler surveys. Results will provide current and potential aquaculture producers in the US and Florida with methods for cost effective production of new aquaculture species with well established high demand markets.

Project Methods
Experiments will investigate the use of photo-thermal controlled cycles to off cycle and/or induce pinfish and pigfish to spawn out of season. Cycles will be set on selected time periods which will serve as treatment categories (ex. 120, 150, 180, and 210 days). The efficacy of using different photo-thermal periods to spawn both species will be evaluated based on spawning frequency, number of eggs produced, egg quality (floating vs. sinking eggs, % fertilization, and % hatch), and larval quality (larvae size and 2-3 dph survival). Experiments dosages of hCG, Ovaprim, Ovaplant, and CCPE for the induced spawning of pinfish and pigfish. For each species' hormone experiment, the hormone efficacy will be examined in four separate dosages as suggested in literature for similar species (ex. Ovaprim experiment for pinfish- female injections @ 0.25, 0.5, 1.0, and 2.0 mL/kg). Dose efficacy for each hormone will be determined based on spawning frequency, number of eggs produced, egg quality (floating vs. sinking eggs, % fertilization, and % hatch), and larval quality (larvae size and 2-3 dph survival). After determining the best dosage for each hormone and each species, those dosages will be used to compare the efficacy of each hormone on the induced spawning of each species (ex. Pinfish experiment- female injections @ 0.5 mL/kg Ovaprim, 500 IU/kg hCG, 1.0 pellet/fish Ovaplant, and 10 mg/kg CCPE). Stocking and replication procedures, and efficacy examination will be similar to that described for dosage comparisons. A series of experiments will be designed to determine the optimal light intensity, algal density, photoperiod, salinity, and temperature for larval culture of pinfish and pigfish. Experiments will be designed to examine each environmental factor for each species. Four treatment levels will be used for each factor as suggested in literature for similar species (ex. photoperiod experiment for pinfish- 6L:18D, 12L:12D, 18L:6D, and 24L:0D). Three experiments will be conducted to determine the effects of using nauplii from the copepod (P. pelagicus) or rotifers as food items for larval pinfish, pigfish, and goggle eye. Two treatments (1- all copepod nauplii diet and 2- all rotifer diet) will be used in the experiments for each species. Treatment level efficacy will be evaluated based on larval growth and survival throughout the experiments. An experiment will be conducted to determine the possibility of replacing Artemia with a microparticulate microbound diet for F. Seminolis larval culture. Larvae (0 dph) will be stocked in 15 L tanks at densities of 25 larvae/L. Three feed treatments (1- all decapsulated Artemia diet, 2- combination of Artemia and a microbound diet, and 3- all microbound diet) will be used in the experiment. Diet efficacy will be based on larval growth and survival throughout the experiment. Thirty day old juvenile pinfish and pigfish will be randomly stocked into 120 L culture tanks at 200 fish/m3 and acclimated over a 48h period to one of six salinity levels (0, 5, 10, 15, 20, 25, and 27 g/L) with four replications per treatment. Culture and sampling protocols will also follow the low salinity tolerance pinfish experiment.

Progress 10/01/13 to 09/30/14

Outputs
Target Audience:Current and potential aquaculture producers. Bait retailers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest?Presentations at national conferences What do you plan to do during the next reporting period to accomplish the goals?more reporting of market for marine bait

Impacts
What was accomplished under these goals? I) defined spawning protocols for pigfish IV) evaluated markets for culture marine baitfish

Publications

• Type: Journal Articles Status: Published Year Published: 2014 Citation: DiMaggio, M.A., C.L. Ohs, J.S. Broach, T.D. Sink. 2014. Effects of stocking density on growth, survival, and stress physiology of pigfish. North American Journal of Aquaculture 76:201-210.

Progress 10/01/12 to 09/30/13

Outputs
Target Audience: Current and potential aquaculture producers, state and federal officials, faculty and extension agents, sea grant agents, teachers, general public Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Four extension workshops were conducted with association of Florida Sea Grant agents and participants included current and potential producers, state extension and research faculty, sea grant agents, and state agency representatives. How have the results been disseminated to communities of interest? Four extension workshops were conducted with association of Florida Sea Grant agents. Extension documents and complied folders of information were distributed to participants. Websites were used to disseminate general information about marine baitfish aquaculture. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Ballyhoo H. balao spawning substrate preference experiments were conducted with two populations and three spawning substrates (foam, PVC, and plastic cable ties). A total of 266 spawning events were recorded with 142,822 eggs collected. Analysis of substrate preference revealed cable ties were the preferred spawning substrate with a mean of 160 eggs collected from each spawning event compared to foam with 60 and pvc with 39 eggs per spawning event. Pigfish eggs exhibited hatching success over 80% during static incubation at densities up to 1,000 eggs/L. Flow-through seawater allowed densities to be increased to 4,000 eggs/L without any observed effects on hatching success. Pinfish were less tolerant to higher densities during static incubation and hatching success declined significantly at densities above 250 eggs/L. Flow-through seawater allowed pinfish densities to be increased to 1,000 eggs/L and was critical to hatching success. Spawning activity of pinfish was not induced at any dose of channel catfish pituitary extract (CCPE) administered. In contrast, CCPE induced spawning for pigfish at all doses tested, although 10 or 20 mg/kg maximized viable spawning frequency and viable egg production. Egg and larval quality of pigfish spawns obtained from CCPE 10 or 20 mg/kg doses were only slightly less than that of a control treatment (0.5 mL/kg of Ovaprim®). Results from multiple pigfish larval experiments indicated rotifer enrichment with DHA Protein Selco and green water application using live T-ISO at 500,000 cells/mL were essential for high survival of pigfish during early stages of larval culture. A once daily rotifer feeding regime at a high concentration of 20 rotifers/mL and a stocking density of 50 larvae/L were also beneficial in maximizing survival. Observations of natural seasonal spawning in pigfish revealed brood have the capacity to spawn consistently over a 100 day period and yield an absolute fecundity over 560,000 eggs/female. Incorporation of krill into brood diet resulted in increases in egg and larval quality. An invitation to participate in an online survey was emailed to 240,000 marine recreational fishing license holders. A total of 14,871 respondents completed some portion of the questionnaire for a 16.1% response rate; with 10,642 anglers completing the entire survey. Of the respondents, 87.5% were male and 70.5% were boat owners. Inshore destinations accounted for the majority of fishing trips taken by Florida anglers. Live bait was caught by 57.8%, purchased by 32.1%, and 10.1% responded they did not use live bait in their fishing activities. The most prominent complaint anglers had with live bait purchased in retails stores was price, followed by availability. Perceptions of aquaculture produced bait were 43.8% positive, 20.6% somewhat positive, and 31% were unknown. Consistent availability and price were identified by anglers as the two most important characteristics for an aquaculture produced marine baitfish.

Publications

• Type: Journal Articles Status: Published Year Published: 2013 Citation: DiMaggio, M.A., J.S. Broach, C L. Ohs. 2013. Evaluation of Ovaprim and human chorionic gonadotropin doses on spawning induction and egg and larval quality of pinfish, Lagodon rhomboides. Aquaculture 414-415: 9-18.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: C.L. Ohs, M.A. DiMaggio, S.W. Grabe, J.S. Broach, C.A. Watson, N.E. Breen, F.T. Barrows. 2013. The effects of increasing docosahexaenoic acid (DHA) and arachidonic acid (ARA) in brood diets of Monodactylus sebae on fecundity, egg and larval quality, and egg fatty acid composition. North American Journal of Aquaculture 75:285-294.
• Type: Journal Articles Status: Published Year Published: 2013 Citation: DiMaggio, M.A., J.S. Broach, C.L. Ohs, and S.W. Grabe. 2013. Captive volitional spawning and larval rearing of pigfish, Orthopristis chrysoptera. North American Journal of Aquaculture 75:109113.
• Type: Other Status: Published Year Published: 2013 Citation: Ohs, C.L., R.L. Creswell, M.A. DiMaggio. 2013. Growing marine baitfish: A guide to Floridas common baitfish and their potential for aquaculture. Florida Sea Grant SGEB 69. 30 p. (http://www.flseagrant.org/images/PDFs/sgeb69_growing_marine_baitfish.pdf)
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: DiMaggio, M., J. Broach, and C. Ohs. 2013. Evaluation of ovaprim and human chorionic gonadotropin doses on spawning induction and egg and larval quality of pigfish Orthopristis chrysoptera. Aquaculture 2013. Nashville, TN. p. 276.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Ohs, C., M. DiMaggio, A. Beany, and C. Adams. 2013. Evaluation of recreational angler perception and demand for cultured marine baitfish in Florida. Aquaculture 2013. Nashville, TN. p. 801.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Broach, J., C. Ohs, M. DiMaggio, G. Wallat, and S. Grabe. 2013. Effects of egg incubation density on larval quality and morphometrics for pinfish Lagodon rhomboides and pigfish Orthopristis chrysoptera. Aquaculture 2013. Nashville, TN. p. 134.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Broach, J., C. Ohs, M. DiMaggio, A. Beany, A. Palau, and S. Grabe. 2013. Effects of commercial rotifer enrichments and rotifer feeding density during larval culture of pigfish Orthopristis chrysoptera. Aquaculture 2013. Nashville, TN. p. 135.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: DiMaggio, M., J. Broach, C. Ohs, and T. Sink. 2013. Effect of stocking density on growth, survival, and stress physiology of pigfish Orthopristis chrysoptera. Aquaculture 2013. Nashville, TN. p. 275.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: DiMaggio, M., J. Broach, and C. Ohs. 2013. Evaluation of ovaprim and human chorionic gonadotropin doses on spawning induction and egg and larval quality of pinfish Lagodon rhomboides. Aquaculture 2013. Nashville, TN. p. 274.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Ohs, C., A. Palau, M. DiMaggio, and J. Broach. 2013. Effects of various salinities and temperatures on hatching success and survival of larval ballyhoo Hemiramphus balao. Aquaculture 2013. Nashville, TN. p. 800.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: DiMaggio, M., C. Ohs, A. Palau, and J. Broach. 2013. Evaluation of culture techniques and spawning substrates for ballyhoo Hemiramphus balao. Aquaculture 2013. Nashville, TN. p. 273.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Garr, A., S. Laramore, C. Ohs, and B. Myers. 2013. Aquaculture in action part 1: Evaluation of the teach aquaculture curriculum. Aquaculture 2013. Nashville, TN. p. 391.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Ohs, C., L. Creswell, M. DiMaggio, J. Broach, A. Beany, and G. Wallat. 2013. Culture methods for commercially viable marine baitfishes in the southeastern United States. Aquaculture 2013. Nashville, TN. p. 802.
• Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: OMalley, P.O., A. Palau, C. Ohs, L. DAbramo, and C. Green. 2013. Feeding larval gulf killifish Fundulus grandis: Utilizing a transitional feeding regime of live and artificial feeds. Aquaculture 2013. Nashville, TN. p. 429.

Progress 10/01/11 to 09/30/12

Outputs
OUTPUTS: Results were presented at four county extension meetings. Four presentations were given at a national conference, Aquaculture America 2012 in Las Vegas, NV. PARTICIPANTS: Matthew DiMaggio - graduate student UF Jason Broach - graduate student UF Audrey Beany - biologist UF TARGET AUDIENCES: Current and potential aquaculture producers. Scientists and extension agents. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
F. seminolis is euryhaline and can survive through a salinity transfer from 0 to16 g/L using NaCl and up to 24 g/L using natural seawater. Regardless of salinity source, acute transfer to 32 g/L resulted in 100% mortality. Using a gradual salinity acclimation, a survival rate of 100% was achieved when salinity was changed from 0 to 32 g/L over 24, 48, 72, and 96 hours. F. seminolis subjected to a gradual salinity increase over 46 days exhibited a maximum tolerance of 78 g/L with a calculated LC-50 of 60 g/L. To determine the effects of salinity on reproduction, broodfish were acclimated to various salinities and spawning success was measured. Egg production, fertilization, and hatching success of F. seminolis decreased with an increase in salinity. Fertilization of eggs spawned above 21 g/L was below 50 % and was not successful at 34 g/L. Sperm extracted from fish acclimated to various salinities of water were not motile at salinities greater than 21 g/L. Pigfish, 3 female and 3 male, were given Ovaprim injections at 0.5 mL/kg and 0.25 mL/kg, respectively, every 48 hours during 70 d. Sixty-one spawns were obtained during the experiment with a total egg production of 1,266,313 eggs. The mean fertilization % of all floating eggs produced (1,115,039 eggs) was 98%, which also had a mean hatch % and 2 dph larval survival of 87.7% and 64.2%, respectively. Both the hatch % and 2 dph larval survival declined throughout the study. The mean fatty acid % levels for DHA, EPA, and AA of floating eggs were 15.17, 2.83, and 0.44, respectively. The 2 dph larval survival was positively correlated to the EPA:AA ratio. Captive pinfish were induced to spawn with a single injection of Ovaprim. Successful culture of pinfish larvae revealed first feeding to occur at 3 days post hatch (DPH) and established a successful larval feeding regime using rotifers, Brachionus plicatilis, and Artemia. Larvae were observed to have completed swim bladder inflation by 19 DPH, with 100% of larvae exhibiting notochord flexion by the same date. Larval survival ranged from 10-117% through 33 DPH. Results from Ovaprim and human chorionic gonadotropin (HCG) dose evaluation experiments showed low dose Ovaprim (0.25, 0.50 mL/kg) and 4000 IU/kg HCG to be the most advantageous for successful spawning induction of pinfish. A fecundity of 1,147,149 eggs/female (2,959 eggs/g body weight) was calculated from 58 volitional spawning events in captive pigfish. Larval survival ranged from 6.5-100.0% through 25 days post hatch (DPH). Larvae were observed to initiate swim bladder inflation by 9 DPH, with 100 % of larvae exhibiting notochord flexion by 19 DPH. Spawning assessments with pigfish indicated superior performance and increased egg and larval quality with the 0.50 mL/kg Ovaprim dose. Spawning reliability was questionable with all HCG doses. Pigfish stocking density experiments revealed similar trends in growth and survival for tested densities. Specific growth rate estimates ranged from 2.02-4.38% for both experiments. Densities up to 0.5 fish/L appear conducive to captive culture although higher densities should be investigated to possibly reduce intraspecific aggression.

Publications

• Cassiano, E.J., C.L. Ohs, M.A. DiMaggio. 2012. Stress survival in larvae of Florida pompano (Trachinotus carolinus) fed enriched rotifers (Brachionus plicatilis) and nauplii of the calanoid copepod (Pseudodiaptomus pelagicus). The Israeli Journal of Aquaculture - Bamidgeh 64:793-800.
• Cassiano, E.J., C.L. Ohs, C.R. Weirich, N.E. Breen, and A.L. Rhyne. 2011. Evaluation of larval Florida pompano, Trachinotus carolinus, fed nauplii of the calanoid copepod Pseudodiaptomous pelagicus. North American Journal of Aquaculture 73: 114-123.
• Ohs, C. L., L.M.V. Onjukka, M.A. DiMaggio, J.S. Broach. 2012. Current Status of the culture of the Seminole killifish Fundulus seminolis as a freshwater and marine baitfish. Aquaculture America 2012. Las Vegas, NV. p. 345.
• Broach, J.S., M.A. DiMaggio, C.L. Ohs, A.H. Beany, N.E. Breen, A.L. Rhyne, S.M. DeSantis, S.W. Grabe. 2012. Egg and larval characteristics from repeated induced volitional spawns of pigfish Orthopristis chrysoptera. Aquaculture America 2012. Las Vegas, NV. p. 73.
• DiMaggio, M.A., C.L. Ohs, A.H. Beany, C.M. Adams. 2012. Evaluation of market potential for aquacultured marine baitfish in Florida. Aquaculture America 2012. Las Vegas, NV. p. 126.
• DiMaggio, M.A., C.L. Ohs, J.S. Broach, S.W. Grabe. 2012. Captive volitional spawning and larval rearing of pigfish Orthopristis chrysoptera. Aquaculture America 2012. Las Vegas, NV. p. 127.