Aquaculture 283 (2008) 6876

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Aquaculture
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / a q u a - o n l i n e

Effects of dietary lipid source and concentration on channel catsh (Ictalurus punctatus) egg biochemical composition, egg and fry production, and egg and fry quality
Todd D. Sink , Rebecca T. Lochmann
School of Agriculture, Fisheries and Human Sciences, University of Arkansas at Pine Bluff, Pine Bluff, AR 71601, USA

a r t i c l e

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a b s t r a c t
We conducted a study to determine the effects of lipid source (poultry fat, PF or menhaden sh oil, FO) and concentration (4 or 10% lipid supplementation) in channel catsh broodstock diets on subsequent egg biochemical composition, egg production, and egg and fry quality. Three female and one male USDA-103 strain three-year-old broodsh were stocked into 16 3860-L outdoor tanks in January and randomly assigned one of the four diets (four replicates per diet). Fish were fed based on a water-temperature-dependant schedule. Spawning cans were added to tanks when water temperature reached 21 C, and the cans were examined three times weekly for egg masses. Masses were enumerated, weighed, measured, analyzed for proximate and fatty acid composition, and incubated. Date to hatch, hatching success, fry growth and survival, and ngerling growth and survival were monitored. Supplementation of catsh broodstock diets with 10% sh oil increased spawning success, fecundity, total egg volume (matrix removed), individual egg weight, eggsspawn 1 (volumetric), total egg lipid concentration, hatching success, and fry survival compared to a control diet with 4% sh oil. The 10PF diet performed similarly to the 10FO diet in all measured parameters except egg fatty acid composition, although results for the 10PF diet did not always differ from those of the 4FO or 4PF diets. The 10FO diet produced signicant decreases in egg saturates and MUFA, and increases in n3 HUFAs and the ratio of n3:n6 fatty acids compared to all other diets. However, in sh fed the 10FO or 10PF diets the differences in egg fatty acid composition did not result in different rates of hatching success, fry survival, or ngerling production. Improved spawning performance of catsh fed 10%-lipid diets may be due partly to increased dietary energy relative to the 4%lipid diets. Although the dietary n3:n6 fatty acid ratios of the 10PF (0.3) and 10FO (5.6) diets were quite different, similar concentrations of HUFAs such as DHA and AA were present in the eggs of sh fed either diet. This indicates that catsh can synthesize HUFAs from 18-carbon precursors and deposit the HUFAs in the egg, and that a variety of lipid sources may be suitable for use in catsh broodstock diets. Current methods of channel catsh egg and fry production using growout diets with 2832% protein and 67% lipid are suboptimal based on the results of this study. 2008 Elsevier B.V. All rights reserved.

Article history: Received 3 March 2008 Received in revised form 15 July 2008 Accepted 16 July 2008 Keywords: Channel catsh Broodstock Eggs Fry Lipid Fatty acids

1. Introduction The basic nutrient requirements for market size channel catsh Ictalurus punctatus are well known (NRC, 1993), but diet research for catsh broodstock is lacking compared to other sh production industries. Brood channel catsh are typically fed the same diets used for ngerling and food sh production (Robinson and Li, 1996, 2002). A USDA survey found that 21.8 2.5 (SE)% of producers fed a 28% protein

Abbreviations: SE, standard error of the mean; 4FO, 4% menhaden sh oil diet; 4PF, 4% poultry fat diet; 10FO, 10% menhaden sh oil diet; 10PF, 10% poultry fat diet; DO, dissolved oxygen; h, hours; min, minutes; ANOVA, analysis of variance; AA, arachidonic acid; MUFA, monounsaturated fatty acids; HUFA, highly-unsaturated fatty acids. Corresponding author. Tel.: +1 870 575 8174; fax: +1 870 575 4639. E-mail address: tsink@uaex.edu (T.D. Sink). 0044-8486/$ see front matter 2008 Elsevier B.V. All rights reserved. doi:10.1016/j.aquaculture.2008.07.024

diet, 59.3 2.9% of producers fed a 32% protein diet, and 13.3 2.2% of producers fed a 35% protein diet (USDA, 2003). Commercial 28 and 32% protein diets typically contain 47% lipid, but production diets may not contain optimal nutrient levels for channel catsh broodstock. Currently, no separate nutrient requirements have been established for channel catsh broodstock, and existing production diets are used due to convenience. Only 3040% of females spawn each year across the channel catsh industry, so producers hold 23 times the minimum weight of broodsh to ensure that egg production goals are met (Steeby and Avery, 2005). This method may not optimize spawning success or the production of high-quality eggs and fry. Lipid manipulation of channel catsh broodstock diets offers the best chance of improving egg and fry quality. Lipids (fats, fatty acids) are important dietary components for sh reproduction and fry survival. Lipid mobilization from the liver and muscle to developing gonads in

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white seabream, Diplodus sargus (Perez et al., 2007), as well as changes in fatty acid composition in broodsh gonad tissues of gilthead seabream (Jerez et al., 2006), suggest that dietary lipid intake and subsequent broodsh lipid composition play crucial roles in reproductive maturation and gonadal development in sh. Increases in egg quality, fecundity, and fertilization success have been linked to dietary highly-unsaturated fatty acids (HUFA) in sh (Izquierdo et al., 2001). Highly-unsaturated fatty acids also affect steroidogenesis in sh (Izquierdo et al., 2001). Arachidonic acid (20:4n6; AA) stimulates reproductive hormone production (Wade et al., 1994) and courtship behavior (Cole and Smith, 1987) in some species of sh. Both n3 and n6 fatty acids are potentially essential for all sh (Sargent et al., 2002), and channel catsh require 1 to 2 g100 g diet 1 of linolenic acid (18:3n3) or 0.5 to 0.75 g100 g diet 1 of eicosapentaenoic acid (20:5n3; EPA) and/or docosahexaenoic acid (22:6n3; DHA); (Satoh et al., 1989a,b). Information on the effects of dietary lipid composition on gamete and fry production and quality could reduce the number of broodsh needed to meet production goals and facilitate broodstock-diet development for channel catsh to increase egg and fry production. The use of regionally available lipids such as poultry fat may reduce costs, while producing a comparable or better-quality product compared to non-sustainable marine sh oils. Poultry fat, which is rich in 18:2n6, may be suitable for catsh broodstock diets. However, sources of n3 fatty acids such as sh oil may be more critical in broodstock diets than in production diets, and very little marine sh oil is used in production diets. Freshwater omnivorous sh, such as channel catsh, may not require a dietary source of n3 highlyunsaturated fatty acids due to their ability to elongate and desaturate 18-carbon fatty acids (such as 18:2n6 and 18:3n3) to n3 and n6 HUFAs (El-Sayed et al., 2005). However the fatty acid requirements of broodstock catsh for optimal reproduction have not been evaluated. The relative importance of n3 and n6 fatty acids for reproduction can be assessed qualitatively by supplementing diets with lipids differing in fatty acid composition. Therefore, we conducted a feeding and spawning trial to determine the effects of lipid source (poultry fat or menhaden sh oil) and concentration (4 or 10% lipid supplementation) of channel catsh broodstock diets on subsequent egg biochemical composition, egg production, and egg and fry quality. 2. Methods 2.1. Experimental system The experimental system consisted of 16 (2.44 m diameter) above ground, outdoor tanks (PolyTank Inc., Litcheld, Minnesota) that were maintained as a ow-through system (10.2 L min 1) with a standing volume of 3860 Ltank 1. Water was supplied to each tank from a 3.23 ha groundwater reservoir and supplemental aeration was maintained in each tank. The tanks were open to ambient sunlight and no temperature controls were used, allowing the system to uctuate with ambient air and reservoir temperatures. 2.2. Experimental diets Four diets were formulated to meet or exceed the known nutrient requirements of channel catsh and were extruded as 6.35-mm oating pellets (TestDiet, Purina Mills Inc., Richmond, Indiana) formulated to contain approximately 36% crude protein from plant and animal sources (Table 1). Two diets contained 4% added lipid as poultry fat (PF) or menhaden sh oil (FO) and two diets contained 10% added lipid as PF or FO. The poultry fat was pet grade (Tyson Foods Inc., Springdale, Arkansas) and the menhaden sh oil was Virginia Prime Gold (Omega Protein, Houston, Texas). Isolipidic diets were formulated to contain similar protein: energy ratios. Proximate analysis of the four diets (Table 1) was performed in duplicate to verify concentrations of crude protein (TruSpec N nitrogen analyzer, St. Joseph, Michigan), ash (mufe furnace,

Table 1 Composition (g 100 g dry diet 1) of experimental channel catsh broodstock diets Dieta 4FO (control) Ingredient (g 100 g dry diet ) De-hulled soybean meal Ground whole-corn Wheat middlings Cottonseed meal Menhaden sh meal Menhaden sh oil Poultry fat preserved with ethoxyquin Pyridoxine hydrochloride Di-calcium phosphate Vitamin and mineral premixb Actual composition (g100 g dry diet 1)c Crude protein Lipid Ash Dry matter Moisture Energy (kcal g 1)d Proteind Lipidd Carbohydratesd
1

4PF 50.61 15 10.37 10 8 4 0.922 0.614 0.487 36.1 6.4 6.8 97.5 2.5 1.442 0.559 1.439

10FO 54.3 15 1.21 10 8 10 0.1 0.901 0.487 36.4 12.1 6.7 96.5 3.5 1.457 1.066 1.242

10PF 54.3 15 1.21 10 8 10 0.1 0.899 0.487 36.5 12 6.8 97.1 2.9 1.457 1.057 1.246

50.73 15 10.37 10 8 4 0.723 0.693 0.487 36.3 6.5 6.7 97.1 2.9 1.442 0.563 1.438

a Diet abbreviations are as follows: 4FO = 4% menhaden sh oil; 4PF = 4% poultry fat; 10FO = 10% menhaden sh oil; 10PF = 10% poultry fat. b Vitamin and mineral premix contains (g g 100 g dry diet 1): menadione dimethylpyrimidinol bisulte, 0.150; L-ascorbyl-2-polyphosphate, 0.057; trace mineral mix, 0.050; folic acid, 0.030; cholecalciferol, 0.030; sodium selenite, 0.025; thiamin mononitrate, 0.021; riboavin, 0.020; DL-alpha tocopheryl acetate, 0.018; calcium pantothenate, 0.017; biotin, 0.015; ethoxyquin, 0.013; vitamin A acetate, 0.012; nicotinic acid, 0.011; choline chloride, 0.010; cyanococobalamin, 0.005; zinc oxide, 0.002; and DL-methionine, 0.001. c Values are means of three replicate samples per diet. d Diet analysis provided by the manufacturer (TestDiet, Purina Mills Inc., Richmond, Indiana).

600 C for 3 h), dry matter (dry matter oven, 130 C for 2 h); (AOAC, 1984), and total lipid (chloroform/methanol method); (Folch et al.,1957). The 4FO diet was the control because its analyzed lipid composition (6.5% total lipid; 61.5% of total lipid from sh oil) is similar to that of commercial growout diets recommended for channel catsh broodstock .The analyzed protein concentration for the control (36.3% crude protein) and other diets (36.136.5% crude protein) was above the minimum concentration recommended (28 or 32% crude protein; Robinson and Li, 1996) to ensure that protein was not a limiting factor in egg and fry production. The fatty acid composition of the diets (Table 2) was determined using the total lipid extracts of the diets that were transestered with boron triouride. Fatty acid methyl esters were quantied using a gas chromatograph equipped with a fused silica capillary column (15 m0.25 mm inside diameter; Varian CP select for FAME #CP8510) and a ame ionization detector (Varian 3380, Varian, Inc., Palo Alto, California). Fatty acids were identied by comparison of retention times to those of known standards (GLC-96, GLC-473B, Nu-Chek Prep, Elysian, Minnesota) and expressed as g 100 g total fatty acids 1. 2.3. Broodsh and feeding schedule Brood channel catsh were selected based upon recommendations by Steeby and Avery (2005) and Steeby et al. (2006). They were three-yearold sh (age 4 at time of spawning) from a registered, genetically improved line (USDA-103) and were selected based upon best growth from that age class. Broodstock (N250) were maintained in a 0.04-ha pond and fed a commercial oating diet with 32% crude protein (Arkat Mills, Inc., Dumas, Arkansas) according to temperature-dependant feeding recommendations for broodstock (Robinson and Li, 1996) prior to the spawning trial. The broodsh were seined, sexed by examination of urogenital opening and genital papilla, weighed, and stocked at the end of January 2007 to allow sh to acclimate to the tanks prior to gamete maturation. Channel catsh broodstock commonly are stocked at a sex ratio of 1:1 or 2:3 males to females (Steeby and Avery, 2005). We chose a slightly

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Table 2 Select fatty acid composition (g 100 g total fatty acids 1) of channel catsh broodstock diets used to evaluate parental lipid intake effects on egg and fry production Fatty acidsa Dietb 4FO (control) 14:0 16:0 18:0 Saturatesc 16:1 18:1d 20:1 MUFAe 18:2n6 18:3n3 20:4n6 20:5n3 22:5n3 22:6n3 n3 HUFAf n6 HUFAg n3/n6 ratio 4.2 14.4 18.6 4.3 8.7 1.2 14.2 19.1 4.1 17.6 23.7 41.3 2.4 4PF 1.1 15.7 4.7 21.5 2.6 21.8 24.4 40.5 5.2 3.1 5.3 8.4 0.3 10FO 5.1 14.2 19.3 4.9 7.4 2.4 14.7 8.0 2.6 1.3 20.2 3.5 26.2 49.9 1.3 5.6 10PF 1.0 15.4 5.4 21.8 2.9 26.2 29.1 37.1 4.3 2.1 3.7 5.8 0.3

can insertion was recorded and later averaged to determine the mean date of spawning. 2.5. Egg processing and enumeration In the hatchery egg masses were acclimated to well water (24.4 25.5 C) for a minimum of ve minutes by mixing and gradual replacement of tank water. After acclimation the entire egg mass (matrix intact) was weighed and recorded. The matrix was dissolved by placing the eggs in a solution of 15 g sodium sulteL water 1 (Isaac and Fries, 1991) for 35 min and the matrix was carefully ushed away with fresh well water using stainless steel screens (1.2 mm mesh) and a catch container to ensure that no eggs were lost. After the matrix was removed, the eggs were placed into a mesh strainer to remove excess water, weighed, and the total egg weight was recorded. The eggs were placed into a 2000-mL graduated pitcher and the total egg volume was recorded. Fifteen milliliters of eggs from each egg mass were then placed into each of three 15-mL labeled polypropylene tubes. The remaining eggs were placed in an incubator to obtain hatching and fry data as described below. Two of the three tubes containing eggs were immediately frozen at 20 C for proximate and fatty acid analyses at a later date. The remaining 15-mL polypropylene tube containing eggs were refrigerated for egg enumeration at the conclusion of egg collection. In the lab, 30 of the previously refrigerated eggs were individually counted, batch weighed, and used to calculate the average individual egg weight. This procedure was repeated twice using different eggs for each count and the means for the two replicates were recorded. These data were used in conjunction with total egg weight (matrix removed) to calculate the number of eggs egg mass 1 by weight by multiplying the total weight of the egg mass by number of eggs g 1. A small sample (13 mL) of eggs was then measured to the nearest 0.1 mL in a 5-mL graduated cylinder, the volume was recorded, and the number of individual eggs contained within that volume were counted. This procedure was repeated twice using different eggs for each count and the data were used to calculate the number of eggs mL 1 by dividing the number of eggs counted by the total volume of the sample. This data was averaged between the two replicates for each sample and used to calculate the total number of eggs egg mass 1 by volume by multiplying the mean number of eggs mL 1 by the total volume of the eggs (matrix removed). Egg images from each spawn were computer digitized using an image capture analysis system (ICAS, Image Pro Plus, Media Cybernetics Inc., Bethesda, Maryland) to calculate the mean egg diameter. Individual egg images were analyzed for the mean egg diameter with ImageTool software (University of Texas Health Science Center, San Antonio, Texas). The computed egg diameters were used to calculate average individual egg volume for each egg mass. This data was combined with mean individual egg weights to calculate average individual egg density for each egg mass. The recorded number of spawns per tank and the egg enumeration data were also used to calculate total fecundity per female, total spawns and eggs produced per tank and diet, and percentage of females that spawned per diet treatment. Fecundity was calculated as the volumetrically determined total number of eggs produced in a tank divided by the total nal weight of
Table 3 Modied temperature-dependant feeding schedule for channel catsh broodstock fed diets with different concentrations of poultry fat or menhaden sh oil Water temperature (C) b10 1016 1621 N21 Feeding rate (g 100 g body weight ) 0.5 1 1 1.5 3 times weekly 3 times weekly 1 time daily 1 time daily
1

a Only fatty acids that comprise greater than 1 g 100 g total fatty acids 1 are presented in the table. b Diet abbreviations are as follows: 4FO = 4% menhaden sh oil; 4PF = 4% poultry fat; 10FO = 10% menhaden sh oil; 10PF = 10% poultry fat. c Saturates included 14:0, 16:0, and 18:0. d Total n7 and n9 isomers. e Monounsaturates included 16:1, 18:1, and 20:1. f Total n3 highly-unsaturated fatty acids included 20:5, 22:5, and 22:6. g Total n6 highly-unsaturated fatty acids included 20:4.

different ratio of 3 females:1 male per tank to increase experimental observations for statistical validity, and to minimize the inuence of territorial aggression in male catsh on results. The 3:1 gender ratio is used by approximately 11.4 1.5% of catsh producers in the United States (USDA, 2003). The four broodsh were stocked into each of sixteen tanks randomly assigned to treatments (4 replicates per diet). Mean (SE) initial weights for the females were 1.403 0.13 kg sh 1 and 1.400 0.09 kg sh 1 for the males. Weight gain of the broodsh was not determined during the study to prevent stress caused by weighing and possible consequences to spawning activity. Broodstock were weighed and counted in each tank at the end of the spawning period to determine weight gain and survival. Each tank was randomly assigned to one of the four experimental diets (four replicates per diet). Commercial feeding practices for catsh broodstock before and during spawning are variable (USDA, 2003), so the sh were fed according to a modied water-temperature-dependant schedule (Table 3) at a frequency high enough to determine diet effects. Water temperature of the tanks was recorded daily, and the diet rations were adjusted accordingly. 2.4. Spawning and egg collection Identical spawning cans (76 61 31 cm surplus military ammunition cans) with 20.3-cm-diameter openings were placed into each tank once water in the tanks reached the initial 21 C (2527 C optimal spawning temperature) spawning temperature (Chapman, 1992) for seven consecutive days (May 7, 2007). The spawning cans were examined for egg masses on Mondays, Wednesdays, and Fridays during the spawning season throughout the months of May, June, and the rst portion of July 2007. Egg collection was concluded after no spawns occurred for ve continuous collection periods (July 11, 2007). Egg masses were carefully removed from the cans by peeling them gently from the edges to ensure removal of the entire mass. Each egg mass was placed into a vented bucket containing water from the same tank, and immediately taken to the hatchery for acclimation and egg processing. Care was taken to minimize the time each egg mass was exposed to air while transfer was taking place. The number of days-post-

Frequency

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2.7. Egg proximate and fatty acid analysis Frozen egg samples were thawed and analyzed for proximate and fatty acid composition. Proximate and fatty acid analysis was performed in duplicate as described for diets (Section 2.2). 2.8. Fry survival and growth Eggs and fry that hatched were left in the incubator for at least 48 h to ensure that all viable eggs had hatched. The fry were then enumerated volumetrically (Steeby and Avery, 2005) to calculate hatching success and fry production. The fry were moved to one of 16 tanks (150 L) that corresponded to experimental replicates of the broodstock tanks in which they were spawned. The tanks were operated as ow-through systems (8.2 L min 1) with well water (23 24 C). The drain standpipe was covered with aluminum screening (3 mm diagonal) to prevent fry escapement. Supplemental aeration was provided in each tank. Each tank was equipped with its own 24-h automatic belt feeder. After the fry began to swim up looking for food, they were offered a fry meal with 48% crude protein and 9% lipid (Arkat Mills Inc., Dumas, Arkansas) at a rate of 25% of body weight daily (Robinson and Li, 1996). The fry were again enumerated volumetrically 14 days-post hatch to determine fry survival during the transition from sac-fry to feeding juveniles. 2.9. Fingerling growth and survival 2.6. Egg hatching One hundred milliliters of eggs (matrix removed) from each egg mass were placed into a labeled compartment of a tray in an 8-tray (24 total segregated compartments) vertical egg incubation system (Marisource, Milton, Washington). Another 100 mL of eggs from the same spawn were placed into a labeled compartment of a different tray. The number of eggs placed into the incubator were later calculated and recorded based on the mean number of eggs mL 1 for that egg mass. These data were used to approximate the hatching success (5%) of eggs and the number of fry produced. Well water (24.425.5 C) was supplied to the incubator at approximately 12 L min 1. The eggs were observed one to two times daily and date of hatching and hatching success were recorded. Fungal growth on the eggs was at times problematic, so eggs with fungal growth were recorded and removed daily, and a prophylactic drip was used every other day to deliver 50 L L 1 hydrogen peroxide (Howe et al., 1999) for control of saprolegniasis. One hundred, 30 to 37 day-old fry produced from each of the 16 broodsh tanks were weighed and stocked into 16 (2.44 m diameter) outdoor tanks with a standing volume of 3860 L tank 1. The water in each tank was maintained as a static system and fertilized with 9.5 L ha 1 of liquid fertilizer (10300 [nitrogenphosphorouspotassium]). Supplemental aeration was maintained in each. The fry were fed 1.5 mm pellets (36% protein, 4% lipid) once daily to satiation for 60 days and then were captured, counted and weighed to determine survival and growth. 2.10. Statistical analysis A two-by-two factorial design and analysis of variance (ANOVA) and descriptive statistics were used to determine differences among treatments using SPSS 11.0 statistical software. Data for percent of females that spawned and survival were arc-sin transformed prior to statistical analysis. Differences were considered signicant at an alpha of 0.05 (P b 0.05), and when differences were found a Tukey's post hoc

Fig. 1. Tank water temperatures during a feeding trial with channel catsh broodstock fed diets differing in lipid source and concentration.

the females in that tank. The nal weight of females in each tank was adjusted to include females that had died during the study to account for total stock production. This was done by adding the number of females that died times the initial mean weight of females in that tank to the total nal female weight.

Table 4 Channel catsh broodstock survival, spawning success, egg production, and physical egg characteristics produced from broodstock fed diets differing in lipid source and concentration Measurement parameter Diet1,2 4FO (control) Total broodstock survival (%) Female broodstock survival (%) Spawning success (spawns100 female brood 1) Fecundity (eggs kg female 1) Spawn time (days-post-can insertion) Water temperature at spawning (C) Egg mass weight, matrix intact (kg) Total egg volume, matrix removed (mL) Individual egg weight (g) Eggs mL 1 Egg diameter (mm) Eggs spawn 1 (volumetric) Eggs spawn 1 (weight) Egg density (g cm3)
1 2

4PF 93.8 6.3 91.5 8.5 50.0 9.6ab 3,976.7 1,229.7ab 27.7 3.5 22.8 0.5 0.347 0.07b 389.2 94.5b 0.0234 0.003b 29.5 3.1 2.4 0.2 12,421.7 3,292.6ab 13,238.2 2,884.9 3.8 0.6

10FO 93.8 6.3 91.5 8.5 83.3 9.6a 7,676.9 418.8a 41.2 3.9 25.0 1.0 0.562 0.04a 678.0 49.5a 0.0282 0.001ab 29.5 1.4 2.2 0.1 19,858.6 1,602.8a 16,121.4 1,304.0 5.7 0.6

10PF 87.5 12.5 83.3 16.8 75.0 16.7ab 6,289.3 1,314.9ab 36.3 4.6 24.2 1.0 0.549 0.05a 690.6 74.4a 0.0306 0.002a 27.8 1.2 2.3 0.1 18.812.2 1,963.2ab 14,412.8 1,201.6 5.1 0.5

81.3 12.0 74.8 16.1 41.6 8.3b 3,149.8 860.9b 29.2 1.4 22.4 0.4 0.376 0.05ab 387.0 67.4b 0.0217 0.002b 31.2 0.7 2.2 0.1 12,062.2 2,140.4b 14,463.1 2,646.2 4.0 0.2

Diet abbreviations are as follows: 4FO = 4% menhaden sh oil; 4PF = 4% poultry fat; 10FO = 10% menhaden sh oil; 10PF = 10% poultry fat. Values are means SE of four replicate tanks per diet. A one-way analysis of variance was used to determine differences among treatment means of the test diets (P b 0.05). When signicant differences were found, treatment means were compared using a Tukey's post hoc test. Means in rows with different superscript letters are different.

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T.D. Sink, R.T. Lochmann / Aquaculture 283 (2008) 6876 Table 6 Select fatty acid composition (g 100 g total fatty acids 1) of channel catsh eggs produced from broodstock fed diets differing in lipid source and concentration Fatty acids Diet1,2 4FO (control) 14:0 16:0 18:0 20:0 Saturates3 16:1 18:14 20:1 24:1 MUFA5 18:2n6 18:3n6 20:2n6 20:3n6 20:4n6 20:5n3 22:4n6 22:5n3 22:6n3 n3 HUFA6 n6 HUFA7 n3/n6 ratio
1

test was conducted to determine specic differences in treatment means. A two-tailed, Pearson bivariate correlation test was conducted to determine which physical and biochemical egg parameters were highly correlated with total egg production, spawning success, egg hatching success, and fry survival to 14 days to determine which variables were associated with increased fry production. 3. Results 3.1. Broodsh The daily water temperatures of the broodstock spawning tanks are shown in Fig. 1. The channel catsh broodstock were fed 119 days of the 145-day study based on the water-temperature schedule (Table 3). Of the 119 days fed, the broodstock received 0.5% of their body weight seven days, 1.0% of their body weight 37 days, and 1.5% of their body weight for 75 days. Mean (SE) initial broodstock weights (females, 1.403 0.13 kg sh 1; males, 1.400 0.11 kg sh 1) were not different among treatments. The female broodstock grew signicantly (P b 0.05) during the seven months they were in the spawning tanks even though a majority of them had spawned. Mean (SE) ending weights for the females was 1.758 0.10 kg sh 1. Males did not gain or lose weight during the study. Seven of 64 broodsh died (10.9%) during the sevenmonth study and the percent broodstock survival was not different among diets (Table 4). Survival was not correlated with spawning success or the number of spawns produced (Pearson correlation coefcient (r) = 0.142). 3.2. Spawning and fecundity At least one egg mass was collected from each broodsh spawning tank and 30 of 48 total possible spawns were collected (62.5% overall spawning success). Broodsh fed the 4FO diet had lower spawning success than sh fed the 10FO diet, while sh fed the 4PF or 10PF diets had intermediate spawning success that was not different from sh fed other diets. Spawning success was strongly positively correlated (Pb 0.05; =0.01; r=0.642) with the concentration of dietary lipid the broodstock received. The mean date of spawning and the mean water temperature at time of spawning (Table 4) were not different among treatments. Fecundity was moderately positively correlated ( = 0.05; r = 0.567) with the dietary lipid concentration. Broodsh fed the 10FO diet had greater fecundity (Table 4) and total egg production tank 1 (40,303.4 887.5 eggs tank 1) than sh fed the 4FO diet (17,880.3 5470.9 eggs tank 1), while sh fed the 4PF (19,857.4 5823.6 eggs tank 1) or 10PF (32,428.8 5967.3 eggs tank 1) diets had intermediate fecundity and total egg production that did not differ from that of sh fed other diets. 3.3. Egg production and physical characteristics Female channel catsh broodstock fed the 10PF or 10FO diets produced heavier egg masses (matrix and eggs; Table 4) than broodstock
Table 5 Composition of eggs produced from channel catsh broodstock fed diets varying in lipid source and concentration Composition (g 100 g eggs 1) Total lipid (dry matter basis) Total lipid (wet basis) Crude protein Ash Dry matter
1

4PF 0.6 0.0 18.5 0.4b 7.7 0.2b 0.6 0.2 27.2 0.4c 3.2 0.3 31.9 0.9c 0.7 0.0ab 1.0 0.1 36.3 0.7b 9.9 2.1 2.4 0.1ab 0.5 0.1 2.9 0.3 7.7 0.7 1.1 0.2b 0.5 0.0 1.6 0.1c 8.3 0.9b 11.0 1.0b 8.2 0.7 0.5 0.1b
b

10FO 1.6 0.2 15.8 0.3a 6.5 0.4a 0.7 0.2 24.5 0.5a 3.1 0.7 22.1 0.5a 0.9 0.1a 0.8 0.1 25.9 1.3a 7.7 1.4 1.4 0.1a 0.6 0.1 2.4 0.2 5.4 1.5 6.5 0.5a 0.5 0.1 1.3 0.1a 19.2 1.8a 27.9 3.7a 7.4 0.3 1.5 0.2a
a

10PF 0.7 0.1b 17.4 0.3b 7.6 0.1b 0.4 0.1 26.1 0.3bc 3.6 0.2 27.4 1.0b 0.7 0.1a,b 0.9 0.5 32.3 1.1b 9.7 0.6 1.6 0.8ab 0.8 0.1 2.9 0.3 7.4 1.0 1.8 0.4b 0.4 0.1 1.6 0.0c 14.3 1.7ab 17.7 2.1b 7.8 1.1 0.8 0.1b

0.5 0.1 17.7 0.4b 7.8 0.2b 0.2 0.0 26.2 0.6bc 3.2 0.3 29.9 1.5bc 0.5 0.2b 1.2 0.1 34.1 1.8b 6.9 0.3 2.6 0.3b 0.7 0.1 3.5 0.2 9.0 0.4 1.3 0.2b 0.5 0.0 1.9 0.1b 11.6 1.5b 14.8 1.5b 9.5 0.3 0.6 0.1b

Diet abbreviations are as follows: 4FO = 4% menhaden sh oil; 4PF = 4% poultry fat; 10FO = 10% menhaden sh oil; 10PF = 10% poultry fat. 2 Values are means SE for eggs from four replicate tanks per diet. Means in rows with different superscript letters are different. 3 Saturates included 14:0, 16:0, 18:0, and 20:0. 4 Total n7 and n9 isomers. 5 Monounsaturates included 14:1, 16:1, 18:1, 20:1, 24:1. 6 Total n3 highly-unsaturated fatty acids included 20:5, 22:5, and 22:6. 7 Total n6 highly-unsaturated fatty acids included 20:4 and 22:4.

fed the 4PF diet, and sh fed the 4FO diet produced egg masses intermediate in weight that were not different from those of sh fed other diets. Broodstock fed the 10PF and 10FO diets produced a greater volume of eggs spawn 1 (matrix removed; Table 4) than broodstock fed the 4PF or 4FO diets. Mean individual egg weights were lower for broodstock fed the 4FO or 4PF diets than for broodstock fed the 10PF diet, while mean individual egg weight for sh fed the 10FO diet was not different from other treatments. The number of eggs mL 1, individual egg volume, and egg diameter (Table 4) from broodstock did not differ among diets. Although the individual egg weights from sh fed the 10PF diet were greater than those of other treatments and the individual egg volume was not signicantly different for all treatments (P N 0.05), the individual egg
Table 7 Hatching success and survival of channel catsh fry produced from broodstock fed diets differing in lipid source and concentration Measurement parameter Diet1,2 4FO (control) First hatch (days)3 Hatching success (sac-fry 100 eggs 1)4 Survival to 14 days (fry 100 sac-fry 1)5 Fry produced (fry 100 eggs 1)6
1

4PF 4.8 0.3 83.3 2.1b 77.5 2.1b 64.6 2.4b

10FO 4.9 0.3 93.5 1.3a 89.0 1.2a 83.3 1.9a

10PF 4.3 0.3 93.9 1.1a 91.7 1.4a 86.1 1.8a

4.4 0.4 84.0 3.7b 74.0 4.3b 62.1 4.1b

Diet1,2 4FO (control) 16.2 2.7b 3.4 0.5 14.1 1.3 1.0 0.2 21.2 0.7 4PF 17.9 2.7b 3.6 0.5 15.4 1.1 1.0 0.0 20.4 0.6 10FO 25.7 2.0a 5.0 0.9 15.1 0.3 0.9 0.2 19.6 0.9 10PF 25.1 2.2a 4.7 0.8 16.2 0.4 1.1 0.2 18.7 0.7

Diet abbreviations are as follows: 4FO = 4% menhaden sh oil; 4PF = 4% poultry fat; 10FO = 10% menhaden sh oil; 10PF = 10% poultry fat. 2 Values are means SE of eggs from four replicate tanks per diet. Means in rows with different superscript letters are different.

Diet abbreviations are as follows: 4FO = 4% menhaden sh oil; 4PF = 4% poultry fat; 10FO = 10% menhaden sh oil; 10PF = 10% poultry fat. 2 Values are means SE of eggs or fry from four replicate tanks per diet. Means in rows with different superscript letters are different. 3 Days-post egg mass collection from spawning tanks. 4 Calculated as estimated (5% interval) sac-fry hatched 100 mL eggs 1 based on calculated number of eggs mL 1 for each individual egg mass. 5 Calculated as number of surviving 14-day-old fry total sac-fry hatched 1. 6 Surviving fry at 14-days-post hatching eggs produced 1.

T.D. Sink, R.T. Lochmann / Aquaculture 283 (2008) 6876 Table 8 Survival and growth rates of fry and ngerlings produced from broodstock fed diets differing in lipid source and concentration Measurement parameter Diet1,2 4FO (control) Fry weight at 14 days of age (g) Fry length at 14 days of age (mm) Fry stocking weight at 3037 days of age (g) Fingerling weight at 60 days-post stocking (g) Fingerlings produced (ngerlings 100 fry stocked 1)
1 2

73

4PF 0.090 0.004 21.6 0.4 0.325 0.014 3.5 0.2b 61.7 4.8b

10FO 0.096 0.006 22.0 0.5 0.334 0.031 5.6 0.7ab 94.8 1.4a

10PF 0.089 0.003 23.5 0.6 0.315 0.021 5.5 0.6ab 92.8 2.1a

0.105 0.005 23.2 0.4 0.311 0.027 6.2 0.9a 30.3 6.6c

Diet abbreviations are as follows: 4FO = 4% menhaden sh oil; 4PF = 4% poultry fat; 10FO = 10% menhaden sh oil; 10PF = 10% poultry fat. Values are means SE of fry or ngerlings from four replicate tanks per diet. Means in rows with different superscript letters are different.

densities (Table 4) were not different among treatments. While the number of eggs spawn 1 was not different among sh fed different diets when determined by mass (Table 4), the number of eggs spawn 1 was signicantly different among diets when determined volumetrically. Mean number of eggs spawn 1 was greater for broodstock fed the 10FO diet than sh fed the 4FO diet, while the number of eggs spawn 1 for broodsh fed the 10PF or 4PF diets were not different from those of sh fed other diets. 3.4. Proximate and fatty acid composition of eggs There were no differences in crude protein, dry matter, ash, or lipid (wet basis) concentration of eggs among treatments (Table 5). Eggs from broodsh fed the 10FO or 10PF diets had a greater lipid (dry matter basis) concentration than eggs from broodsh fed the 4FO diet, while lipid concentration in eggs from sh fed the 4PF diet were not different from the control. Fatty acid analysis (Table 6) was calculated using the concentration of individual fatty acid total egg lipid (wet basis) 1 and reported as g fatty acid 100 g fatty acids 1. Eggs from broodstock fed the 10FO diet contained less saturated and monounsaturated (MUFA) fatty acids than eggs from broodstock fed the 4FO, 4PF, or 10PF diets (Table 6). Eggs from broodstock fed the 10FO diet contained greater concentrations of total n3 fatty acids and n3 highly-unsaturated fatty acids (HUFA; b18 carbons and four or more double bonds) than eggs from broodstock fed the 4FO, 4PF, or 10PF diets (Table 6). There were no differences in total n6 fatty acids or n6 HUFA concentrations in eggs among treatments. Eggs from broodstock fed the 10FO diet also contained less n9 MUFA than eggs from broodstock fed the 4FO, 4PF, or 10PF diets (Table 6). The total n3:n6 ratio was signicantly greater (3 times; P b 0.05) in eggs from broodstock fed the 10FO diet than in eggs from broodsh fed other diets. 3.5. Fry hatching success and survival There was no difference in the number of days-post collection to rst hatch among the diets (Table 7). Eggs from broodsh fed the 10FO or 10PF diets exhibited greater hatching success, fry survival to 14 days, and total fry produced100 eggs 1 (Table 7) than eggs from broodstock fed either the 4FO or 4PF diets. Fry hatching success was strongly, positively correlated with individual egg weight (Pb 0.01; =0.01; r=0.523), egg total lipid content (Pb 0.01; =0.01; r=0.700), egg n3 HUFA content (Pb 0.01; =0.01; r=0.542), egg n3:n6 HUFA ratio (Pb 0.01; =0.01; r=0.515) and moderately, positively correlated with individual egg density (P b 0.05; = 0.05; r = 0.365). Hatching success of the eggs increased with increasing egg weight, total lipid content, n3 HUFA content, n3:n6 HUFA ratio, and egg density. Fry hatching success was strongly, negatively correlated with egg saturated fatty acid concentration (Pb 0.01; =0.01; r=0.510) and MUFA concentration (Pb 0.01; =0.01; r=0.534). Fry survival to 14 days was strongly, positively correlated with individual egg weight (P b 0.01; = 0.01; r = 0.776), egg total lipid concentration (P b 0.01; = 0.01; r = 0.763), egg n3 HUFA concentra-

tion (P b 0.01; = 0.01; r = 0.507), and egg n3:n6 HUFA ratio (P b 0.01; = 0.01; r = 0.487). Fry survival to 14 days was strongly, negatively correlated with egg MUFA concentration (P b 0.01; = 0.01; r = 0.497) and moderately, negatively correlated with egg saturated fatty acid concentration (P b 0.01; = 0.05; r = 0.451). 3.6. Fry and ngerling growth There were no differences in weight or length of fry at 14 days or weight at time of stocking (3037 days of age) among fry from broodstock fed any of the diets (Table 8). Fingerlings from broodsh fed the 4FO diet were heavier than ngerlings from broodsh fed the 4PF diet, and the weight of ngerlings from broodsh fed the 10PF or 10FO diets were not different from ngerlings from sh fed other diets (Table 8). Fingerlings from broodsh fed the 10PF or 10FO diets had higher survival and more ngerlings were produced100 fry stocked 1 (Table 8) than ngerlings from sh fed the 4PF or 4PF diets. Fingerlings produced from sh fed the 4FO diet had the lowest survival, which was signicantly lower than that of ngerlings from sh fed the 4PF diet. Fingerling growth was strongly, negatively correlated with ngerling survival (P b 0.01; = 0.01; r = 0.870). 4. Discussion The overall spawning success (61.2%) in this study was much higher than the average success of 40% reported by catsh producers in the United States (Steeby and Avery, 2005), and broodsh mortality (10.9%) was lower than the industry average of 14% (USDA, 2003; Steeby and Avery, 2005). The overall spawning success was likely inuenced by the greater spawning success of broodsh fed the 10% lipid diets (10PF, 75.0%; 10FO, 83.3%), as broodsh fed the control (4FO) diet had spawning success (41.6%) similar to the industry average. Producers typically use low-fat (67% lipid in 32% crude-protein diets) diets such as the 4PF or 4FO diets, but spawning success in this study increased as dietary lipid concentration increased. Fecundity and total egg production tank 1 were higher for broodsh fed the 10FO diet compared to the control diet, but sh fed the 10PF diet had similar results to those fed the 10FO diet. Regardless of lipid source, dietary lipid concentration was strongly, positively correlated with increased egg production female 1, indicating that higher-lipid commercial diets could signicantly increase production per female and reduce the number of broodstock needed to meet egg production goals. Broodsh fed the 10%-lipid diets produced heavier egg masses (10PF, 0.549 kg mass 1; 10FO, 0.562 kg mass 1) than broodsh fed the 4%-lipid (4FO, 0.376 kg mass 1; 4PF, 0.347 kg mass 1) diets. The average weight of egg masses produced from broodsh fed any diet was lower than the 0.617 0.05 kg mass 1 reported as the industry average (USDA, 2003), and is likely due to the young age (4 years at time of spawning) of the broodstock. Fish fed the 10FO or 10PF diets produced a greater volume of eggs spawn 1 than sh fed the 4PF or 4FO diets although the number of eggs mL 1 and egg diameters were not different for eggs from broodsh fed different diets. These facts combined with the great disparity between the number of eggs spawn 1 by volume and by weight indicate

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T.D. Sink, R.T. Lochmann / Aquaculture 283 (2008) 6876

that there was a great deal of variation in individual egg volume and weight within egg masses. The lipid composition of eggs is amenable to manipulation via the maternal diet (Watanabe, 1982) and biochemical composition of eggs is often used as an indicator of egg quality. However, the relationship between biochemical composition and egg quality is difcult to interpret (Morehead et al., 2001). In this study, concentration of total lipid in eggs and the fatty acid composition of the eggs had a profound effect on hatching success of eggs and survival of fry. Typical hatching rates for channel catsh eggs are 7690%, with a few hatcheries reporting greater than 90% hatching success (USDA, 2003). The hatching success for eggs from broodsh fed the 4FO or 4PF diets was within this range. However, eggs from broodstock fed either the 10FO or 10PF diets had higher total lipid concentration and greater hatching success and fry survival than eggs from broodstock fed 4FO or 4PF diets. Increased egg lipid concentration is vital for fry survival as lipids are the primary energy source used by sh from fry to maturity (Sargent et al., 1989). The 10FO diet and eggs from broodsh fed the 10FO diet were lower in saturates and MUFAs than eggs from sh fed diets with poultry fat. Saturated fatty acids and MUFAs are important sources of metabolic energy for broodstock (Sargent et al., 2002). Based on the hatching success and survival of fry from broodsh fed the 10FO diet, saturates and MUFAs do not appear to be as critical to egg and fry survival as n3 and n6 fatty acids. However, the eggs from broodsh fed the 10FO diet contained higher concentrations of total lipid, and this may have compensated for the differences in concentrations of saturates and MUFAs available to the embryos and fry for metabolic energy. Poultry fat contains high concentrations of oleic acid (18:1n9), which is an important energy source during development of marine sh larvae (Van der Meeren et al., 1991), and is required for normal development of sh embryos (Fernande-Palacios et al., 1997). This could explain why eggs and fry from broodsh fed the 10PF diet performed similarly in a number of parameters compared to those from broodsh fed the 10FO diet. Poultry fat is less expensive than sh oil, and it is likely that a combination of poultry fat and sh oils would be effective in channel catsh broodstock diets to supply larval metabolic energy and enhance embryo development. The superior quality of eggs produced by broodsh fed the 10FO or 10PF diets resulted in better-quality fry. Results with FO are similar to those obtained with many marine sh (Izquierdo et al., 2001; Sargent et al., 2002), but our results indicate that PF is just as effective in broodstock diets for channel catsh when a higher supplemental concentration (10%) of lipid is used. During egg development sh fry absorb and retain fatty acids in the same ratio as found in the egg yolk, which is determined largely by the maternal diet (Hayes et al., 1973). Increased egg quality, fecundity, fertilization success, and egg hatching success in sh have been linked to dietary n3 HUFA concentration (Izquierdo et al., 2001; Sargent et al., 2002). Fish eggs often contain more n3 than n6 fatty acids (Leger et al., 1981; Eldridge et al., 1983; Tocher and Sargent, 1984), but there is less data on freshwater species. Eggs from catsh broodsh fed the 10FO diet were highest in total n3 fatty acids and n3 HUFAs. Eggs from broodstock fed any of the other diets were higher in n6 fatty acids than n3 fatty acids. However, good egg hatching success and fry survival resulted from eggs with n3 to n6 ratios of 0.8 to 1.5, while signicantly reduced performance occurred in eggs with n3 to n6 ratios of 0.50.6. A dietary n3 to n6 fatty acid ratio of 1.5 resulted in good broodstock performance and egg quality in another study with catsh broodstock (SRAC, 2006). However, our results indicate that ratios below this are also effective, which would reduce the need to add marine sh oils to catsh broodstock diets. Eicosapentaenoic acid (20:5n3) and DHA (22:6n3) were preferentially retained in eggs from broodsh fed the 10FO diet, which suggests that n3 HUFAs have a signicant role in development of channel catsh eggs. However, no pronounced reproductive benets of high n3 HUFA concentrations in the 10FO diet (50 g 100 g fatty acids 1) or 10FO eggs (27 g 100 g fatty

acids 1) were observed relative to the concentrations in the 10PF diet (6 g 100 g fatty acids 1) or 10PF eggs (18 g 100 g fatty acids 1). Despite statistical differences in total n3 HUFAs of eggs from sh fed 10FO or 10PF diets, the concentration of DHA was similar and above 14 g 100 g 1 in eggs of sh fed either diet. The total n3 HUFAs in eggs of sh fed 4FO or 4PF were also lower than that of the 10FO eggs, but the concentration of DHA in eggs of sh fed the 4FO or 4PO diets was less than 12 g 100 g 1. The differences in DHA concentrations were numerically small in some cases, but essential fatty acid requirements for most sh range from 0.52.0% of the diet (NRC, 1993). The concentration of DHA in the 4FO diet appeared to be insufcient to support hatching success and fry survival equivalent to that obtained with the 10FO or 10PF diets. The concentration of DHA may be a better indicator of egg quality than total n3 HUFA due to the critical role of DHA in cell membranes and neural tissues (Sargent, 1995). Although freshwater omnivorous sh, such as channel catsh, do not require a dietary source of HUFAs for growth due to their ability to elongate and desaturate 18carbon fatty acids to HUFAs (El-Sayed et al., 2005), requirements for reproduction may differ. The 10PF diet contained enough 18:3n3 to result in high DHA concentrations in the egg (similar to the 10FO diet), but the 4PF diet did not. Therefore, the potential of channel catsh fed diets without marine sh oils to produce eggs high in DHA may only be realized when sufcient concentrations of dietary 18:3n3 are present. A 2:1 ratio of DHA: EPA has also been observed commonly in sh eggs (Sargent, 1995), suggesting biological signicance and implications for quality. However, the egg EPA concentration in catsh did not follow the same pattern as DHA and was only elevated in eggs of sh fed the 10FO diet. The ratio of DHA: EPA in eggs ranged from 39 and did not correlate with hatching success or fry survival. Identifying optimal ratios of specic fatty acids for different functions may be less relevant for omnivorous sh such as channel catsh that can bioconvert 18-C fatty acids into HUFAs. In contrast to the n3 fatty acids, the concentrations of n6 fatty acids such as 18:2n6 and AA were similar in eggs of sh irrespective of diet. The 6-desaturase enzyme has a greater afnity for 18:3n3 than 18:2n 6, but the higher concentrations (8 fold) of 18:2n6 than 18:3n3 in the poultry fat diets likely favored desaturation of 18:2n6 by competitive inhibition of 18:3n3 (Balfry and Higgs, 2001). Most of the diets contained no detectable AA, but the concentration of 18:2n6 was sufcient for synthesis and deposition of at least 5 g 100 g fatty acids 1 of AA in the eggs. Arachidonic acid concentration is often maintained within narrow limits due to its critical roles in eicosanoid synthesis and membrane function (Fountoulaki et al., 2003). Spawning success was higher in sh fed the 10FO diet compared to sh fed the control diet, and the 10FO diet was the only diet that contained a detectable concentration of AA. Arachidonic acid stimulates reproductive hormone production (Wade et al., 1994) and courtship behavior (Cole and Smith, 1987) in some species of sh. However, sh fed the 10PF diet had similar spawning success to those fed the 10FO diet, so dietary AA did not appear to be critical for channel catsh to achieve spawning success. The intent of this study was to improve egg and fry production relative to the 4FO control diet, which is similar a commercial formula. No statistical differences in spawning parameters of sh fed the 10FO or 10PF occurred during this study except for egg fatty acid composition. However, the 10FO diet improved spawning success and fecundity compared to the control diet when the 10PF diet did not, and some of the differences in performance on the 10FO and 10PF diets may have biological and possibly economic signicance. Using data from this study, we generated an example of biological signicance in reference to production from 100 2-kg female broodsh: (1) Control (4FO) diet: (100 females 41.6% spawning success) [(200 kg female body weight3150 eggs kg body weight 1)(84% hatching success)(74% fry survival)]=162,909 fry produced (2) 10PF diet: (100 females 75.5% spawning success) [(200 kg female body weight 6289 eggs kg body weight 1) (93.9% hatching success) (91.7% fry survival)] = 817,699 fry produced

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75

(3) 10FO diet: (100 females 83.3% spawning success) [(200 kg female body weight 7677 eggs kg body weight 1) (93.5% hatching success) (89.0% fry survival)] = 1,064,310 fry produced The difference in fry production between the 10FO and 10PF diets per 100 2-kg females is 246,611 fry, which appears to be a minute difference when millions of fry are produced. However, this difference in fry production is 1.5 times the entire amount of fry produced from 100 2-kg females fed the control diet and the total fry production of the 10FO diet represents a 6.5-fold increase in fry production compared to a 5-fold increase for the 10PF diet. Both 10PF and 10FO diets statistically improved reproduction compared to the control diet, but biologically, the 10FO diet improved egg and fry production compared to the 10PF diet. These biologically signicant results could affect production economics, but suitable economic models must be developed to test this. Although there were no differences in growth of fry from broodstock fed different diets, the ngerlings produced from broodsh fed the control (4FO) diet grew to a much larger size. This is likely due to the high mortality rates in ngerlings produced from broodsh fed 4%-lipid diets, resulting in reduced density and less competition for food in the ngerling tanks. As ngerling density within the tanks decreased, more natural food (zooplankton and insects) resources were available to the remaining ngerlings for growth. It is unclear how the physiological effects mediated by lipids in the parental diets affect growth or survival of catsh offspring. Increased dietary energy, increased available energy (from lipid), or both appeared to enhance survival of ngerlings from broodsh fed diets with 10% added lipid, rather than the specic fatty acid composition of the diets. However, it is important to note that all diets met the n3 HUFA requirement of channel catsh (0.50.75% of the diet). 5. Conclusions Supplementation of catsh broodstock diets with 10% sh oil increased spawning success, fecundity, total egg volume (matrix removed), individual egg weight, eggs spawn 1 (volumetric), total egg lipid concentration, hatching success, and fry survival compared to a control diet with 4% sh oil. The 10PF diet performed similarly to the 10FO diet in all measured parameters except egg fatty acid composition, although results for the 10PF diet did not always differ from those of the 4FO or 4PF diets. The 10FO diet produced signicant decreases in egg saturates and MUFA, and increases in n3 HUFAs and the ratio of n3:n6 fatty acids compared to all other diets. However, in sh fed the 10FO or 10PF diets the differences in egg fatty acid composition did not result in different rates of hatching success, fry survival, or ngerling production. Improved spawning performance of catsh fed 10%-lipid diets may be due partly to increased dietary energy relative to the 4%lipid diets. Although the dietary n3:n6 fatty acid ratios of the 10PF (0.3) and 10FO (5.6) diets were quite different, similar concentrations of HUFAs such as DHA and AA were present in the eggs of sh fed either diet. This indicates that catsh can synthesize HUFAs from 18-carbon precursors and deposit the HUFAs in the egg, and that a variety of lipid sources may be suitable for use in catsh broodstock diets. Current methods of channel catsh egg and fry production using growout diets with 2832% protein and 67% lipid is suboptimal based on the results of this study. Broodstock performance could be improved using diets supplemented with 10% sh oil, poultry fat, or a combination, effectively reducing the number of broodsh needed to meet the egg and fry production goals of the channel catsh industry. Acknowledgment We thank the Arkansas Catsh Promotion Board for funding this project. Technical support was provided by Nick Kinsey and Harold Phillips.

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