Dietary lipid level but not L-carnitine affects growth performance of hybrid striped bass ž Morone chrysops Y=M. saxatilis?/

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1 Aquaulture Dietary lipid level but not L-arnitine affets growth performane of hybrid striped bass ž Morone hrysops Y=M. saxatilis?/ T.G. Gaylord, D.M. Gatlin III ) Department of Wildlife and Fisheries Sienes and Faulty of Nutrition, Texas A&M UniÕersity System, College Station, TX, , USA Reeived 31 January 2000; reeived in revised form 26 April 2000; aepted 26 April 2000 Abstrat The objetives of this experiment were to determine if dietary lipid levels in exess of previously reported minima would inrease performane of hybrid striped bass, and to determine if supplementation of L-arnitine would improve growth andror body omposition when feeding elevated dietary lipid. Therefore, a 2=4 fatorial design was utilized to test the effiay of dietary L-arnitine at 0 and 3000 mgrkg diet at eah of four lipid levels Ž 5%, 10%, 15% and 20%.. Semipurified diets were formulated to ontain 40% rude protein and dextrin was substituted for menhaden oil at a rate of 2.25 to 1 to maintain digestible energy at approximately 14.2 kjrg. Juvenile reiproal ross hybrid striped bass initially averaging 2.5 grfish were ultured in a 5 brakish water reirulating system and fed twie daily at a rate approahing apparent satiation. At the end of the 8-week feeding trial, L-arnitine supplementation did not influene weight gain, but dietary lipid level did, with the lowest value Ž 1084% of initial weight. ahieved by fish fed 5% lipid and the highest Ž 1343%. by fish fed 15% lipid; fish fed 20% lipid had intermediate weight gain Ž 1215%.. Feed effiieny also was influened by dietary lipid with fish fed the 10% and 20% lipid diets having higher feed effiieny values than those fed the 5% and 15% lipid diets. Body ondition indies were only slightly altered by L-arnitine supplementation, but dietary lipid over 10% influened body ondition. Liver omposition was altered by dietary lipid level but not L-arnitine supplementation. As dietary lipid level inreased, liver lipid inreased and liver glyogen dereased dramatially, espeially in fish fed the diet with 20% lipid, whih had no soluble arbohydrates. Musle omposition was unaltered by any dietary treatment. In onlusion, ) Corresponding author. Tel.: q ; fax: q address: d-gatlin@tamu.edu D.M. Gatlin r00r$ - see front matter q 2000 Elsevier Siene B.V. All rights reserved. PII: S X

2 238 ( ) T.G. Gaylord, D.M. Gatlin IIIrAquaulture hybrid striped bass generally did not utilize dextrin as effiiently as lipid, and lipid levels between 10% and 15% of diet provided maximum growth with intermediate lipid deposition. q 2000 Elsevier Siene B.V. All rights reserved. Keywords: L-Carnitine; Lipid; Hybrid striped bass; Morone; Nutrition 1. Introdution Lipid nutrition of fish produed in aquaulture has attrated onsiderable interest both historially from the standpoint of satisfying essential fatty aid requirements ŽNRC, and more reently for its protein-sparing apability. Current salmonid diets may ontain upwards of 30% lipid for enhaning feed effiieny as well as growth of these oldwater arnivorous speies Ž Hardy, Most warmwater speies are typially fed diets with less than 10% lipid due to their ability to utilize higher levels of arbohydrate for energy, the high ost of lipid relative to arbohydrate, and unwanted aretion of lipid depots. Various warmwater speies also may differ in arbohydrate utilization aording to natural feeding habits, with more arnivorous speies typially having redued ability to utilize dietary arbohydrate Ž Wilson, Hybrid striped bass are rosses of two arnivorous Morone speies, whih generally would not be expeted to utilize arbohydrates for energy as effiiently as lipid. One previous study indiated hybrid striped bass were able to utilize dextrin as effiiently as lipid for energy based on weight gain and feed effiieny Ž Nematipour et al., 1992a.. However, more reent studies have indiated these fish may use lipid more effiiently than arbohydrate for energy Ž Huthinson et al., 1998; Gaylord and Gatlin, in press.. To improve utilization of dietary lipids by fish, L-arnitine has shown promise by improving growth and feed effiieny and reduing lipid deposition in some speies Table 1 Composition of the basal diets Ž gr100 g dry weight b. a Dietary lipid % Speial Selete menhaden fishmeal Casein b Menhaden oil Dextrin d Vitamin premix d Mineral premix Carboxymethyl ellulose Cellulose a Diets were formulated to ontain 40% rude protein and either 5%, 10%, 15% or 20% lipid with 14.2 kj estimated digestible energy per g diet. Diets at eah lipid level were supplemented with L-arnitine ŽUS Biohemial, Cleveland, OH. at either 0 or 3000 mgrkg. b Omega Protein, Reedville, VA. US Biohemial, Cleveland, OH. d Same as Moon and Gatlin Ž

3 ( ) T.G. Gaylord, D.M. Gatlin IIIrAquaulture ŽSantulli and D Amelio, 1986; Torreele et al., 1993; Chatzifotis et al., 1995; Ji et al., Previous researh in this laboratory, however, did not find any benefits of dietary L-arnitine supplementation at relatively low lipid levels for hybrid striped bass ŽGaylord and Gatlin, in press.. Therefore, the purpose of the present study was to determine if hybrid striped bass ould utilize elevated levels of dietary lipid, and if supplemental L-arnitine ould enhane utilization of dietary lipid. 2. Materials and methods Diets were formulated to ontain 40% rude protein and were maintained isoalori at an estimated digestible energy level of 14.2 kjrg Ž Nematipour et al., 1992b. by adjusting dextrin and menhaden oil levels Ž Table 1.. All other known nutritional requirements of hybrid striped bass were satisfied by these diets Ž Gatlin, A fatorial design was utilized in whih diets ontaining lipid at either 5%, 10%, 15% and 20% were supplemented with L-arnitine at 0 and 3000 mgrkg diet. Lipid levels were hosen to meet Ž 5% or 10%. or exeed Ž 15% and 20%. previously reported optima for hybrid striped bass Ž Nematipour et al., 1992a.. The rather high L-arnitine level was Table 2 Weight gain, feed effiieny and musle L-arnitine onentration of hybrid striped bass fed diets ontaining four levels of lipid at two L-arnitine levels Dietary treatment Weight gain Feed effiieny Musle L-arnitine Lipid % L-Carnitine mgrkg Ž % of initial weight. Ž g gainrg dry feed. Ž mgrg. a,b a a 0.60a 6.64a a 0.55A 11.60A b 0.73b 6.82a b 0.56AB 20.50B a 13.50b B 30.90C b 0.70b 16.20b b 0.57AB 61.20D ANOVA, Pr ) F d Lipid L-Carnitine Ž 0) Ž Lipid=L-Carnitine Pooled S.E a Values are means of three repliate groups. b Average initial fish weight was 2.5 g. Means of three fish from eah of three repliate groups expressed on a fresh-weight basis. d Signifiane probability assoiated with the F-statisti. If a signifiant Ž P main effet ourred with signifiant interation, lipid levels within an L-arnitine level followed by different letters Ž0 L-arnitines lower ase; 3000 L-arnitinesupper ase. are signifiantly different as determined by Dunan s multiple range test; whereas, a signifiant main effet without interation allowed data to be pooled by lipid level prior to means separation.

4 240 ( ) T.G. Gaylord, D.M. Gatlin IIIrAquaulture hosen beause levels up to 1000 mgrkg did not affet growth or body omposition of hybrid striped bass in a previous study Ž Gaylord and Gatlin, in press.. L-Carnitine was added to eah diet by dissolving in water and mixing prior to pressure pelleting, ensuring this hygrosopi ompound was evenly distributed in the diets. Other proedures for diet preparation and storage were as previously desribed ŽMoon and Gatlin, Juvenile reiproal ross hybrid striped bass Ž Morone hrysops Y = M. saxatilis?. were obtained from a ommerial produer Ž Keo Fish Farm, Keo, AR. and were maintained in freshwater earthen ponds. Prior to the experiment, fish were moved indoors into a brakish water Ž 5. reirulating system ontaining 24, 38-l aquaria maintained at 27"28C and on a 12:12 h diurnal yle. Water flow into the aquaria was restrited to 1 lrmin. The fish were onditioned to the system and fed the basal diet with 5% lipid for 2 weeks prior to the start of the experiment. Ten hybrid striped bass initially averaging 2.5 grfish at the start of the 8-week experiment were stoked into eah aquarium Ž tripliate aquaria per dietary treatment. and fed their respetive diets twie daily a perentage of their body weight to a level approahing apparent satiation. Feeding rates were adjusted weekly to ensure limited feed waste as well as supplying Table 3 Body ondition indies of hybrid striped bass fed diets ontaining four levels of lipid at two L-arnitine levels a b d Dietary treatment HSI IPF ratio MR Lipid % L-Carnitine mgrkg b 2.24a 43.5b b 2.64A 44.6b b 44.4b BC 43.7b b 5.13b 40.6a b 5.13C 41.6a a a a 4.50B 41.1a ANOVA, Pr ) F e Lipid L-Carnitine Lipid=L-Carnitine Pooled S.E a Values are means of three fish from eah of three repliate groups. b Hepatosomati index Ž HSI. sliver weight=100rbody weight. Intraperitoneal fat Ž IPF. ratiosipf weight=100rbody weight. d Musle ratio Ž MR. sfillet weight=100rbody weight. e Signifiane probability assoiated with the F-statisti. If a signifiant Ž P main effet ourred with signifiant interation, lipid levels within an L-arnitine level followed by different letters Ž0 L-arnitines lower ase; 3000 L-arnitinesupper ase. are signifiantly different as determined by Dunan s multiple range test; whereas, a signifiant main effet without interation allowed data to be pooled by lipid level prior to means separation.

5 ( ) T.G. Gaylord, D.M. Gatlin IIIrAquaulture ample food for maximized growth. At the termination of the trial, three fish per aquarium were randomly removed, euthanized, and hepatosomati index ŽHSI s liver weight= 100rfish weight., intraperitoneal fat Ž IPF. ratio ŽIPF weight= 100rfish weight. and musle ratio Ž MR s fillet weight= 100rfish weight. were determined. Musle and liver samples from these three fish per aquarium also were removed and stored at y208c prior to proximate analysis that followed standard proedures ŽAOAC, Musle and liver total lipid was determined by hloroformrmethanol extration Ž Folh et al., Analysis of free L-arnitine in musle and diets was performed aording to the method of Wieland et al. Ž Musle and liver lipid lass analysis was performed using the Iatrosan TLC-FID system Ž Biosan, Washington, DC. aording to the proedures of Toher et al. Ž with modifiations of the solvent system Ž Craig and Gatlin, Glyogen was determined in the liver aording to the method of Hassid and Abraham Ž Statistial analysis of the data was performed using fatorial analysis of variane Ž SAS, Differenes were onsidered signifiant at P F If interations ourred between main effets then differenes among treatments were determined by Dunan s multiple-range test Ž Dunan, Table 4 Liver and musle omposition of hybrid striped bass fed diets ontaining four levels of lipid at two L-arnitine levels a b b b Dietary treatment Liver lipid % Liver glyogen % Musle lipid % Lipid % L-Carnitine mgrkg a a a a b 12.50b b 12.71b a a 4.31 ANOVA, Pr ) F Lipid L-Carnitine Lipid=L-Carnitine Pooled S.E a Values are means of three fish from eah of the three repliate groups. b Expressed on a fresh-weight basis. Signifiane probability assoiated with the F-statisti. If a signifiant Ž P main effet ourred with signifiant interation, lipid levels within an L-arnitine level followed by different letters Ž0 L-arnitines lower ase; 3000 L-arnitinesupper ase. are signifiantly different as determined by Dunan s multiple range test; whereas, a signifiant main effet without interation allowed data to be pooled by lipid level prior to means separation.

6 242 ( ) T.G. Gaylord, D.M. Gatlin IIIrAquaulture Results Diets not supplemented with L-arnitine were analyzed to ontain undetetable levels of free L-arnitine while 85%, 98%, 92% and 93% of the targeted level of 3000 mg L-arnitinerkg was analyzed in the diets ontaining 5%, 10%, 15% and 20% lipid, respetively. Musle L-arnitine levels were readily modified by dietary supplementation of L-arnitine and were also affeted by dietary lipid onentrations Ž Table 2.. A notable interation ourred with high levels of dietary lipid augmenting L-arnitine supplementation in elevating musle free L-arnitine levels. Dietary fators had dramati effets on weight gain and feed effiieny Ž Table 2.. Fish fed diets ontaining 5% lipid grew less than the fish fed the other lipid levels; however, L-arnitine had no effet on weight gain at any level of dietary lipid. Feed effiieny was generally improved by inreasing dietary lipid. L-Carnitine supplementation also affeted feed effiieny of hybrid striped bass with fish fed the L-arnitine-supplemented diets having redued feed effiieny ompared to fish fed the same level of dietary lipid without L-arnitine supplementation. Body ondition indies were influened by dietary L-arnitine supplementation and lipid level Table 3. Inreasing dietary lipid up to 20% redued HSI values but elevated Table 5 Liver and musle lipid lasses of hybrid striped bass fed diets ontaining four levels of lipid at two L-arnitine levels a,b a,b Dietary treatment Liver lipid lasses % Musle lipid lasses % Lipid Ž %. L-Carnitine TG FFA CE PL TG FFA CE PL Ž mgrkg ab a ab a a b a b a b a b b b b b ANOVA, Pr ) F Lipid L-Carnitine Lipid=L-Carnitine Pooled S.E a Values are means of three fish from eah of the three repliate groups. b Relative perentages of total identified lipid lasses: triglyerides Ž TG., free fatty aids Ž FFA., holesterol esters Ž CE., phospholipids Ž PL.. Signifiane probability assoiated with the F-statisti. If a signifiant Ž P main effet ourred with signifiant interation, lipid levels within an L-arnitine level followed by different letters Ž0 L-arnitines lower ase; 3000 L-arnitinesupper ase. are signifiantly different as determined by Dunan s multiple range test; whereas, a signifiant main effet without interation allowed data to be pooled by lipid level prior to means separation.

7 ( ) T.G. Gaylord, D.M. Gatlin IIIrAquaulture IPF ratio values. The IPF ratio was redued by L-arnitine supplementation in fish fed 20% lipid. MR was unaltered by L-arnitine supplementation but was signifiantly redued at lipid levels in exess of 10% of diet. Liver lipid was inreased in response to dietary lipid supplementation Ž Table 4.. A moderate elevation in liver lipid ourred in fish fed diets with 15% lipid, although the greatest elevation in liver lipid ourred in fish fed diets with 20% lipid. Liver glyogen onentrations were inversely related to liver lipid Ž Table 4.. As dietary lipid inreased above 10%, glyogen onentrations in the liver dereased, most dramatially when dietary lipid was elevated from 15% to 20%. Musle lipid was not affeted by either dietary lipid or L-arnitine supplementation Ž Table 4.. The relative amounts of triglyerides, free fatty aids, holesterol and holesterol esters in musle lipids were not signifiantly affeted by either of the dietary fators Ž Table 5.. Liver lipid lasses were not affeted by dietary L-arnitine supplementation; however, liver triglyerides were depressed at an inlusion level of 20% dietary lipid Ž Table 5.. Cholesterol esters in liver lipid were redued as dietary lipid inreased above 5%. Relative amounts of free fatty aids and phospholipid in liver were not altered by either of the dietary fators. 4. Disussion The findings in this study are onsistent with those of Huthinson et al. Ž and Gaylord and Gatlin Ž in press. in whih hybrid striped bass had improved weight gain and feed effiieny when dietary lipid was elevated from 2% or 5%, respetively, to levels of 11% and 10%, respetively. In these studies diets were estimated to be isoalori by substituting dextrin for lipid at a rate of 2.25 to 1 by weight. An earlier study Ž Nematipour et al., 1992a. reported suh a substitution rate resulted in similar growth performane of hybrid striped bass fed diets with lipid levels ranging from 2% to 10%. However, the more limited energy effiay of dietary arbohydrates relative to lipid for hybrid striped bass also is supported by the study of Rawles and Gatlin Ž In that study hybrid striped bass were not able to digest soluble arbohydrates as effetively as lipid. In the urrent study, lipid also appeared to provide more energy for metabolism of hybrid striped bass, resulting in inreased growth rates as well as enhaned feed effiieny. At least 10% dietary lipid should be present to support the most rapid weight gain of hybrid striped bass. A dietary lipid level of 15% appeared to provide slightly exessive energy as determined by inreased deposition of lipid in the liver and peritoneal avity as well as depressed musle yield. Similar results have been demonstrated with red drum, a arnivorous marine siaenid, showing a preferential use of lipid rather than dextrin for energy when dietary lipid was inreased from 3% to 10% with a onomitant derease in dextrin Ž Serrano et al., The inrease in dietary lipid improved weight gain, feed effiieny, as well as protein effiieny of red drum aross three different dietary protein levels. Relative hanges in growth and feed utilization as well as lipid deposition in the body should give a reasonable indiation of when dietary energy is adequate or exessive. Although no energy determinations were performed on whole fish from the present

8 244 ( ) T.G. Gaylord, D.M. Gatlin IIIrAquaulture experiment, a fatty fish of greater or equal size has stored more energy than smaller, leaner fish. This, taken along with a slight improvement in feed effiieny appears to indiate improved utilization of dietary lipid for net energy stores by hybrid striped bass ompared to the soluble arbohydrate, whih was used to keep diets isoalori. Therefore, the maximal dietary lipid energy that an be utilized by hybrid striped bass for lean mass aretion an be roughly extrapolated from the urrent study. It appears that the omposition of growth was not altered greatly until the diet provided between 10% and 15% lipid. A dietary energy supply of up to 3760 kjrkg diet from lipid was benefiial for hybrid striped bass growth when they were fed a 40% rude protein diet. However, when dietary lipid energy was elevated to 5648 kjrkg diet, lipid aretion inreased relative to weight gain. A minimum level of soluble arbohydrate in the diet does appear to be benefiial to hybrid striped bass as the diet that was devoid of soluble arbohydrate Ž 20% lipid. showed slightly depressed weight gain. Similar trends have been shown with other arnivorous fish speies. Rainbow trout have been shown to require low levels of soluble arbohydrate to maximize growth through minimizing protein degradation rates Ž Peragon et al., MGoogan Ž also observed depressed weight gain of red drum fed an isoalori diet devoid of soluble arbohydrate. Thus, a minimum level of soluble arbohydrate in the diet appears neessary to maximize growth and feed effiieny of arnivorous fish, but the urrent study was not designed to address a minimum level of soluble arbohydrate for maintaining maximal growth. The limited influene of L-arnitine on hybrid striped bass growth and feed effiieny has been noted in another study from this laboratory Ž Gaylord and Gatlin, in press. as well as with other speies of fish Ž Rodehutsord, 1995; Chatzifotis et al., A similar outome ourred in the present study even with L-arnitine supplementation at three times the level fed to hybrid striped bass in the previous study. The only notable benefit of L-arnitine supplementation ame in the redution of IPF deposition in fish fed the diets with 20% lipid, though the redution was relatively small. There appeared to be a slight Ž P s derease in liver lipid deposition with L-arnitine supplementation in fish fed diets with 15% and 20% lipid. Other researhers have noted more dramati influenes of dietary L-arnitine on omposition of growth. For example, Ji et al. Ž noted that Atlanti salmon fed diets supplemented with L-arnitine had redued fillet lipid. Red sea bream also have been shown to be responsive to L-arnitine supplementation. Chatzifotis et al. Ž reported inreased growth rate and feed effiieny when L-arnitine was supplemented at a level of 1087 mgrkg diet. The lak of a strong L-arnitine effet on metabolism in the present study appears to be due to the ability of fish to synthesize adequate quantities of L-arnitine for lipid metabolism. The diets were not limiting in lysine or methionine, whih are preursors for L-arnitine synthesis. If a limited preursor pool was available for metabolism, supplemental L-arnitine may have had more dramati influenes on growth andror omposition of gain. Lipid supplementation also appeared to elevate L-arnitine synthesis by hybrid striped bass. An inrease in dietary lipid above 10% dramatially inreased levels of musle free L-arnitine. The mehanism by whih this ours is not lear, though it is reasonable to postulate that more L-arnitine may be needed by the fish for metabolism

9 ( ) T.G. Gaylord, D.M. Gatlin IIIrAquaulture of dietary lipid. However, in the urrent study fish fed diets with and without L-arnitine supplementation had equal performane and omposition. Lipid lass alterations in response to dietary L-arnitine supplementation were not observed in the urrent study. These findings are similar to those of a previous study in whih arnitine supplementation up to 1000 mgrkg diet was ineffetive in altering lipid utilization by hybrid striped bass Ž Gaylord and Gatlin, in press.. In that study, there also was no influene of dietary lipid level on lipid lass profile in the liver or musle. In the urrent study, similar finding were observed with dietary lipid levels of up to 15%, but when dietary lipid was elevated to 20% a notable effet was observed in relative perentages of liver triglyerides and holesterol esters. One might have expeted an inrease in triglyerides as dietary lipid inreased beause triglyerides are the primary storage form of lipid, but when dietary lipid was 20% liver triglyerides dereased. The derease in liver triglyerides appeared to be due to lipolysis in that a small, but not quite signifiant Ž P s 0.10., elevation in free fatty aids ourred. The relative perentage of holesterol esters in liver lipid also was modified by level of dietary lipid. Cholesterol esters appeared to be diluted out as dietary lipid inreased above 5%. This would be reasonable beause triglyerides and free fatty aids were the most prominent forms of lipid in the liver. Although limited effets were observed as dietary lipid levels inreased, it appears that extremely high lipid diets may alter the metabolism of lipids in the liver. This may be detrimental to the fish as dietary lipid levels of 20% inreased liver lipid and redued growth. In onlusion, data from the present study appear to indiate a limited benefit of L-arnitine supplementation to hybrid striped bass fed diets with lipid ranging from 5% to 20%. Dietary lipid appears to be more readily utilized as an energy soure than arbohydrate by hybrid striped bass. However, lipid in exess of 10% of diet tended to inrease deposition of lipid in the liver and peritoneal avity. Aknowledgements This researh was supported in part by grant no from the United States Department of Agriulture through the Southern Regional Aquaulture Center. Thanks are extended to Mr. Mike Freeze Ž Keo Fish Farm, Keo, AR. for generously supplying the fish for this study. We also wish to aknowledge Omega Protein for the kind donation of menhaden fish meal Ž Houma, LA. and menhaden fish oil ŽReedville, VA.. Referenes AOAC Ž Assoiation of Offiial Analytial Chemists., Offiial Methods of Analysis. Arlington, VA, 1298 pp. Chatzifotis, S., Takeuhi, T., Seikai, T., The effet of dietary L-arnitine on growth performane and lipid omposition in red sea bream fingerlings. Fish. Si. 61, Chatzifotis, S., Takeuhi, T., Watanabe, T., Satoh, S., The effet of dietary arnitine supplementation on growth of rainbow trout fingerlings. Fish. Si. 63,

10 246 ( ) T.G. Gaylord, D.M. Gatlin IIIrAquaulture Craig, S.R., Gatlin, D.M. III, Growth and body omposition of juvenile red drum Ž Siaenops oellatus. fed diets ontaining leithin and supplemental holine. Aquaulture 151, Dunan, D.B., Multiple range and multiple F tests. Biometris 11, Folh, J., Lees, M., Sloane-Stanley, G.H., A simple method for the isolation and purifiation of total lipids from animal tissues. J. Biol. Chem. 226, Gaylord, T.G., Gatlin, D.M. III, In press. Effets of dietary arnitine and lipid on growth and body omposition of hybrid striped bass Ž Morone hrysops Y= M. saxatilis?.. Fish Physiol. Biohem. Gatlin, D.M. III, Nutrition and feeding of striped bass and hybrid striped bass. In: Harrell, R.M. Ž Ed.., Striped Bass and Other Morone Culture. Elsevier, Amsterdam, pp Hardy, R.W., Problems and opportunities in fish feed formulation. Aquaulture Magazine 25, Hassid, W.Z., Abraham, S., Chemial proedure for analysis of polysaharides. In: Colowik, S.P., Kaplan, N.O. Ž Eds.., Methods of Enzymology vol. 3 Aademi Press, New York, p. 37. Huthinson, C.G., Rawles, S.D., Gatlin, D.M. III, Effets of dietary arbohydrate kind and level on growth, body omposition and glyemi response of juvenile sunshine bass ŽMorone hrysops Y= M. saxatilis?.. Aquaulture 161, Ji, H., Bradley, T.M., Tremblay, G.C., Atlanti salmon Ž Salmo salar. fed L-arnitine exhibit altered intermediary metabolism and redued tissue lipid, but no hange in growth rate. J. Nutr. 126, MGoogan, B.B., Effets of dietary protein and energy manipulations on growth and aspets of nitrogen metabolism of red drum, Siaenops oellatus. PhD dissertation, Texas A&M University, College Station, TX, 135 pp. Moon, H.Y., Gatlin, D.M. III, Total sulphur amino aid requirement of juvenile red drum, Siaenops oellatus. Aquaulture 95, Nematipour, G.R., Brown, M.L., Gatlin, D.M. III, 1992a. Effets of dietary arbohydrate:lipid ratio on growth and body omposition of hybrid striped bass. J. World Aquault. So. 23, Nematipour, G.R., Brown, M.L., Gatlin, D.M. III, 1992b. Effets of dietary energy:protein ratio on growth harateristis and body omposition of hybrid striped bass, Morone hrysops Y= M. saxatilis?. Aquaulture 107, National Researh Counil Ž NRC., Nutrient Requirements of Fish. National Aademy Press, Washington, DC, 112 pp. Peragon, J., Barraso, J.B., Garia-Salguero, L., de la Higuera, M., Lupianez, J.A., Carbohydrates affet protein-turnover rates, growth, and nulei aid ontent in the white musle of rainbow trout ŽOnor- hynhus mykiss.. Aquaulture 179, Rawles, S.D., Gatlin, D.M. III, Carbohydrate utilization in striped bass Ž Morone saxatilis. and sunshine bass Ž Morone hrysops Y= M. saxatilis?.. Aquaulture 161, Rodehutsord, M., Effets of supplemental dietary L-arnitine on growth and body omposition of rainbow trout Ž Onorhynhus mykiss. fed high fat diets. J. Anim. Physiol. Anim. Nutr. 73, Santulli, A., D Amelio, V., Effets of supplemental dietary arnitine on growth and lipid metabolism of hathery-reared sea bass Ž Dientrarhus labrax L... Aquaulture 59, SAS Institute, SAS User s Guide: Statistis, Version. 5th edn., Cary, NC 956 pp. Serrano, J.A., Nematipour, G.R., Gatlin, D.M., Dietary protein requirement of the red drum ŽSiaenops oellatus. and relative use of dietary arbohydrate and lipid. Aquaulture 101, Toher, D.R., Fraser, A.J., Sargent, J.R., Gambel, J.C., Lipid lass omposition during embryoni and larval development in Atlanti herring Ž Clupea harengus L... Lipids 20, Torreele, E., Van Der Sluizen, A., Verreth, J., The effet of dietary L-arnitine on the growth performane in fingerlings of the Afrian atfish Ž Clarias gariepinus. in relation to dietary lipid. Br. J. Nutr. 69, Wieland, O.H., Deufel, T., Paetzke-Brunner, I., Free and esterified arnitine: olorimetri method. 3rd edn. In: Bergmeyer, H.U. Ž Ed.., Methods of Enzymati Analysis vol. VII Verlag Chemie, Deerfield Beah, FL, pp Wilson, R.P., Utilization of dietary arbohydrate by fish. Aquaulture 124,