Animal Production Department, Faculty of Agriculture, Mansoura University, Al-Mansoura, PC , Egypt

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1 Interntionl Journl of Current Reserch in Biosciences nd Plnt Biology ISSN: (Online) Volume 3 Numer 9 (Septemer-2016) Journl homepge: Originl Reserch Article doi: Nutritionl-Reproductive Efficiency Reltionship of Europen Se Bss, Dicentrrchus lrx (Linneus, 1758) Brood Stock: Condition Fctor, Orgns Indices, Antomicl nd istologicl Chrcteristics Emn. Mkld, Ahmed I. Mehrim* nd Mohmed Z.A. Mohmed Animl Production Deprtment, Fculty of Agriculture, Mnsour University, Al-Mnsour, PC , Egypt *Corresponding uthor. A s t r c t A feeding tril ws conducted to study the effect of the live (trsh fish) food (T 1 ), rtificil diet (T 3 ) or mixed etween them (T 2 ) t rte (1:1) on condition fctor (K f ), gondo somtic index (GSI) nd hepto somtic index (SI) prmeters, esides the ntomicl nd histologicl ltertions of the gonds nd the liver of Dicentrrchus lrx femles nd mles rood stock through the feed period t 45 dys, then spwning period t 15 dys. In the feeding period, totl of 180 Se ss rood stock femles nd mles were used. The 90 femles with g per fish s n initil ody weight were seprtely llotted in 9 concert tnks (three tnks / tretment). Also, 90 mles with initil ody weight rnged etween ( g per fish) seprtely llotted in other 9 concert tnks. Fish were stocked t 10 fish per concrete tnk (8 4 1 m) in lck greenhouse in the sme experimentl conditions. The otined results reveled tht oth sex fed the rtificil diet (T 3 ) significntly incresed the K f, GSI, nd SI compred to other dietry tretments (T 1 nd T 2 ). Also, the sme tretment (T 3 ) seriously improved in the ntomicl nd histologicl of the gonds nd the liver mong other dietry tretments. The feeding fish on rtificil diet (T 3 ) hd more effective on ll tested prmeters compred to the trsh fish or the mixed diets, prticulrly for of femles thn mles. Thus from the current findings, it could e concluded tht the useful using of the good qulity rtificil diet in D. lrx htcheries for enhncement their reproductive efficiency. Also these findings confirmed on the positively reltionship etween the feeding sources nd reproductive efficiency of D. lrx rood stock, especilly for femles. A r t i c l e I n f o Accepted: 26 August 2016 Aville Online: 06 Septemer 2016 K e y w o r d s Antomy Feeding Gonds istology Se ss Introduction Europen Se ss, Dicentrrchus lrx is the most importnt commercil nd consumed mrine fish. At present it is the widely cultured in the Mediterrnen re, with Greece, Turkey, Itly, Spin, Croti, nd Egypt eing the lrgest producers (FAO, 2012). Europen Se ss is sesonl reeder, spwning in winter nd erly spring, with some differences in timing ccording to the specific loction. There is only one reproductive seson per yer, which tkes plce in winter in the Mediterrnen popultion (Decemer to Mrch) nd up to June in the Atlntic popultions (Moretti et l., 1999). Fish nutrition is mtter of gret importnce in the expnding Mediterrnen quculture industry s etter product qulity nd optimum growth cn e chieved using the pproprite diets (Prpour nd Alexis, 2001). The min ojective when formulting fish diet is to 27

2 provide nutritionlly lnced mixture of ingredients to support the mintennce, growth, reproduction nd helth of the niml t n ffordle cost (NRC, 1993). As feed is one of the principle costs in feed production (Luptsch et l., 2001), formultions must e sed on our knowledge of nutritionl requirements for them to e economiclly vile. Reproduction in fish, s in other vertertes, is ffected y environmentl, socil nd nutritionl fctors. The effects of food rtion size nd nutrient composition of the diets on reproduction hve een investigted in severl species importnt in quculture (Kjørsvik et l., 1990; Bromge et l., 1992).The qulity nd quntity of food re mong the most importnt exogenous fctors directly ffecting growth nd, indirectly, mturtion nd mortlity in fish, thus eing ultimtely relted to fitness (Wootton, 1990). owever, the effect of the sources of fish feeds on the reproductive efficiency of mrine fish, prtilly D. lrx rood stock is requiring further investigtions. Therefore, the present study ws designed to investigte the effects of the live, rtificil food or mixed etween them t rte (1:1) on condition fctor (K f ), orgns indices prmeters, esides the ntomicl nd histologicl ltertions of the gonds nd the liver of D. lrx mles nd femles rood stock. Mterils nd methods The present study ws performed in El-Sherif privte mrine fish frm, Wdy Mriot, Alexndri Governorte, Egypt. It ws conducted into two periods, the first period is feeding period for 45 dys, then the mturtion rood stock femles injected with 1000 IU hcg (from Argent, US A)/Kg ody weight ccording to Moretti et l. (1999) efore the second period s spwning period for 15 dys. Experimentl procedures Europen Se ss D. lrx mles nd femles rood stock were otined from El-Sherif privte mrine fish frm, Wdy Mriot, Alexndri Governorte, Egypt. Femles nd mles were crefully trnslted to the rering concrete tnks in lck greenhouse. Ech fish sex ws seprtely stocked into the rering concrete tnks for two weeks s n dpttion period. During this period, fish fed diet contining 45% crude protein nd 12% crude ft produced y Aller Aqu for extruded mrine fish feed, Christinsfeld, Denmrk. Concrete tnks (4m 8m 1m) nd liters cpcity in lck greenhouse, continuous ertion ws mintined in ech tnk using n electric ir lower with 7 P 3000 RPM, Chin. The wter ws received in externl reservoir ( m) for sedimenttion, fter tht wter ws filtered, erted using mechnicl nd microil filters, ionizer nd ertor, then wter were pumped to the htchery ponds in the lck greenhouse. Filtrtion ws performed using snd filter. Tnks filled with mrine wter 35 ppt. Wter in ech tnk were chnged twice dy y wter follow system. Wter temperture ws djusted ws mintined t 17 ± 1ºC. Lighting ws performed for 10 hours per dy from 7 m to 5 pm. In the feeding period, Se ss rood stock femles nd mles were distriuted seprtely into six experimentl tretments (s three replictes per tretment) eing three tretments for femles nd other three tretments for femles s shown in Tle 1. Thus, totl of 180 Se ss rood stock femles nd mles were used. The 90 femles with g per fish s n initil ody weight were seprtely llotted in 9 concert tnks (three tnks / tretment). Also, 90 mles with initil ody weight rnged etween ( g per fish) seprtely llotted in other 9 concert tnks Fish were stocked t 10 fish per concrete tnk (4m 8m 1m) in lck greenhouse t the sme experimentl conditions. Fish were fed the trsh fish (srdine nd clmri), which re consider s the common feed for rood stock in Egyptin mrine fish htcheries. Fresh srdine nd frozen clmri were ought from Al-Mx fishing ord, Alexndri Governorte, Egypt. Prior to use, the trsh fish ws crumled to smll pieces s n pproprite size nd stored frozen ( 20 C) until required. The floting rtificil diet ws prepred in the feed fctory t Al-Mx reserch sttion, Ntionl Institute of Ocenogrphy nd Fisheries (NIOF), Alexndri Governorte, Egypt. Artificil diet (48% crude protein nd 15% crude ft) ws prepred to e of the nerly similr chemicl composition of the trsh fish (Tles 2 nd 3). Artificil diet ws extruded s floting pellets, 1 cm in dimeter, nd coted with fish oil fter drying. Fish were fed the trsh fish diet nd their mixing with the rtificil diet (50% : 50%) t rte of 2.5% of their ody weight, while fish fed the rtificil diet t rte of 1% of their ody weight. The feed quntity ws djusted i-weekly ccording to the ctul ody weight chnges. All the experimentl diets mnully introduced to the fish twice dily t 8 m nd 2 pm. 28

3 Tle 1. Detils of the experimentl tretments design in the feeding period. Tretment Detils T 1, Femles fed the trsh fish (srdine nd clmri) s control group T 2, Femles fed the mixing of trsh fish nd rtificil diet (50% : 50%). T 3, Femles fed floting rtificil diet T 4, Mles fed the trsh fish (srdine nd clmri) s control group T 5, Mles fed the mixing of trsh fish nd rtificil diet (50% : 50%). T 6, Mles fed floting rtificil diet Tle 2. Feed ingredients (%) of the rtificil diet used during the feeding period. Ingredient % Fish Mel 999 (72%, CP) Fish Mel (65%, CP) Soy en mel (48%, CP) Corn gluten (60%, CP) Sun flour mel (36%, CP) 8.50 Whet middling (13%, CP) 6.00 Fish oil Whet gluten (80%, CP) 1.00 Animl geltin 0.25 Mono clcium 0.50 Vitmins premix Minerls premix Lysine 0.10 Methionine 0.10 Vitmin C 0.05 Totl Vitmins (per kg feed): Vit. A (1,000,000 IU); Vit. D3 ( IU); Vit. E (100,000 IU); Vit. B1 (5,000 mg); Vit. B2 (10,000 mg); Vit. B6 (10,000 mg); Vit. B12 (10,000 mg); Vit. C (20,000 mg); Biotin (500 mg); folic cid (2,000 mg); pntothenic cid ( mg), nd Vit. K 3 (2,000 mg). 2 Minerls (per kg feed): C CO 3 (314 mg); K 3 PO 4 (469.2 mg); Mg SO O (147.4mg); N Cl, (49.8 mg); Trce elements 19.6 g consists of Fe +2 gluconte; Mn SO 4. 2 O (10900 mg); Zn SO O (3120 mg); Cu SO O (620 mg); KI: (160 mg); Cl CO O; Ammonium molydte (60 mg), nd N 2 SeO 3 (Sodium selente) (20 mg). Tle 3. The chemicl composition of the experimentl diets used during the feeding period. Chemicl composition Artificil diet Srdine Clmri Srdine + Clmri Dry mter (%) Crude protein (%) Crude ft (%) Ash (%) Crude fier (%) 2.10 Nil Nil Nil Nitrogen free extrct (NFE, %) Gross energy (Kcl / 100 g DM) (GE) Protein / energy (P / E) rtio (mg CP / KJ GE) Metolizle energy (ME) 2 (KJ / 100 g DM) GE (KJ / 100 g DM) = CP CF NFE 4.11 clculted ccording to NRC (1993). 2 ME (KJ / 100 g DM) clculted y using fctors of 4.50, 8.10 nd 3.49 Kcl for crude protein, crude ft nd NFE, respectively ccording to Pnth (1982). 29

4 During the feeding period, wter temperture, dissolved oxygen (DO), p nd slinity were monitored twice dily t 15 ml depth using Crison Meters, Spin. While, unionized mmoni ws mesured twice weekly using I Ammoni Low Rnge Portle Photometer nn, Indi. In ll tretments, DO concentrtion rnged from mg/l, p rnged etween ws unionized mmoni concentrtion rnge ws mg/l, Slinity rnge ws 34 ± 2.0 g/l nd temperture rng ws 16 ± 1.0 C. These wter qulity prmeters were ccepted for rering Se ss rood stock ccording to Moretti et l. (1999). Directely fter the feeding period, the spwning period for 15 dys ws eginning, where the cnuol test ws conducted of Europen Se ss rood stock femles in ech tretment to exmine mturity of the eggs (s egg dimeter µm ccording to Moretti et l. (1999), unstuck eggs. Then the mturtion rood stock femles were injected with 1000 IU hcg per kg ody weight (Argent, USA, ccording to Moretti et l., 1999). The concrete tnks were crefully clened efore the spwning period. Then, femles nd mles rood stock of ech tretment (s three tretments) were stocked t 1 : 1 s sex rtio in the clening concrete tnks for 48 hrs. At the end of the feeding period, six fish (n = 6) from ech tretment were rndomly chosen nd nesthetized. Fish were nesthetized y trnsferred in smll plstic tnk contining 10 L wter supplemented with 3 ml pure clove oil dissolved in 10 ml solute ethnol. Then, the rood stock femles nd mles weight nd totl length were tken for the nerest 0.1 g nd 0.01cm, respectively (Fig. 26 & ) to clculte the condition fctor (K f ) ccording to the following eqution; K f = W / L descried y Lgler (1956). Where, W is fish weight (wet weight in g) nd L is fish totl length (in cm). After tht the sme fish were killed nd the dominl cvity ws opened to remove the gonds (mle nd femle) or the liver. Which were individully weighed to clculte the gondo somtic index (GSI) nd hepto somtic index (SI) y the following equtions; GSI = gonds weight 100 / fish weight (Tseng nd Chn, 1982), nd SI = (liver weight / fish weight) 100 (Jngrd et l., 1967). At the strt nd t the end of the feeding period, fish were nesthetized nd scrificed to exmine the development of the ovry nd the sttus of the liver in ll tretments, which were recorded y the digitl cmer. Then, the histologicl chnges in the ovry nd the liver fter the feeding period were checked. The sme fish t the sme numers (n = 6) used for determined the K f, GSI nd SI or the ntomicl ltertions were tken for histologicl exmintion. Fish were nesthetized nd scrificed to otin the ovries nd the livers smples. Smples were fixed in 10% neutrlized formlin solution followed y wshing with t wter, then dehydrted y different grdes of lcohol (70, 85, 96 nd 99%). Smples were clered y xylene nd emedded in prffin wx. The wx locks were sectioned to six micron. The sections were stined y hemtoxyline () nd eosin (E) nd then sujected to histologicl exmintion ccording to Roerts (2001). Sttisticl nlysis Dt of K f, GSI nd SI were reported s men vlues of ll tretments. Replictions (n = 3) nd stndrd errors of men (± SEM) re sed on tnk vlues. All percentges were rcsine-trnsformed prior to sttisticl nlyses. All dt were sujected to one-wy nlysis of vrince (ANOVA) using SPSS for windows (Stndrd version 17.0 SPSS Inc., Chicgo, Illinois, USA). The differences etween men of tretments were compred for the significnce (p 0.05) using Tukey s post hoc significnt test. Results nd discussion Condition fctor (K f ) nd orgns indices prmeters In femles fed the rtificil diet, or fed trsh fish only (srdine nd clmri) T 3 nd T 1, respectively were significntly (p 0.05) incresed the K f compred to the fish fed the mixing of trsh fish nd rtificil diet (50% + 50%, T 2 ). Also, Se ss femle fed the rtificil diet (T 3 ) significntly (p 0.05) incresed GSI compred to fish fed the other dietry tretments (T 1 nd T 2 ). Menwhile, no significnt (p 0.05) differences of SI mong ll dietry tretments (Tle 4). owever, dt indicted tht K f of D. lrx rood stock mles fed the different experimentl diets (T 4, T 5, nd T 6 ) did not significntly (p 0.05) differ fter 45 dys of tretment. Se ss mle fed the trsh fish (T 4 ) reveled tht significntly (p 0.05) incresed SI compred to fish fed the other dietry tretments (T 5 nd T 6 ). Menwhile, no significnt (p 0.05) differences of GSI mong ll dietry tretments (Tle 4). 30

5 Tle 4. Effect of dietry tretments on condition fctor (Kf), gondosmtic index (GSI), nd heptosomtic index (SI) of femle nd mle Se ss (D. lrx) rood stock Tretment Condition fctor (K f ) GSI (%) SI (%) T 1, 1.24± ± ±0.369 T 2, 1.04 ± ± ±0.259 T 3, 1.30 ± ± ±0.525 P- vlue T 4, 1.21± ± ±0.149 T 5, 1.17± ± ±0.006 T 6, 1.21± ± ±0.047 P- vlue Men in the sme column hving different smll letters re significntly different (p 0.05). T 1 nd T 4 : fish fed trsh fish (srdine nd clmri), T 2 nd T 5 : fish fed the mixing of trsh fish nd rtificil diet (50% + 50%), nd T 3 nd T 6 : fish fed floting rtificil diet for femles nd mles, respectively. Antomicl exmintion The ntomicl ltertions of the ovry of femle Se ss (D. lrx) rood stock pre nd fter fed dietry tretments were detected (Fig. 1 d). Where, the dietry tretments (Fig. 1, c, nd d) reveled shrply development of the ovry thn the ovry of fish pre tretments (Fig. 1 ). Prtilly, fish fed the rtificil diet (T 3 ) showed more development of the ovry thn fish fed other dietry tretments (T 1 nd T 2 ). The ntomicl ltertions of the testes of mle Se ss (D. lrx) rood stock pre nd fter fed dietry tretments were detected (Fig. 2 d). Where, the dietry tretments (Fig. 2, c, nd d) reveled shrply development of the testes thn of fish pre tretments (Fig. 2 ). Prtilly, fish fed the rtificil diet (T 6 ) showed more development of the testes thn fish fed other dietry tretments (T 4 nd T 5 ). c Fig.1 d: Showing the ntomicl ltertions of the ovry of femle Se ss (D. lrx) rood stock () pre dietry tretments, nd (, c, nd d) fter fed the trsh fish (srdine nd clmri, T 1 ), fed the mixing of trsh fish nd rtificil diet (50% + 50%, T 2 ), nd fed floting rtificil diet (T 3 ), respectively. d 31

6 c d Fig. 2 d: Showing the ntomicl ltertions of the testes of mle Se ss (D. lrx) rood stock () pre dietry tretments, nd (, c, nd d) fter fed the trsh fish (srdine nd clmri, T 4 ), fed the mixing of trsh fish nd rtificil diet (50% + 50%, T 5 ), nd fed floting rtificil diet (T 6 ), respectively. istologicl chrcteristics The ovry The histologicl ltertions of the ovry of Se ss (D. lrx) rood stock fed different dietry tretments were showed in Figs Microscopiclly, fish fed the trsh fish (srdine nd clmri, T 1 ) showed the ovry with degenerted mture oocytes follicles nd ruptured thecl lyer, s well s very low content of yolk gloules (Fig. 3 ). Fish fed the mixing of trsh fish nd rtificil diet (50% + 50%, T 2 ) showed the ovry with irregulr mture follicles, demturtion thecl lyer nd fir content of yolk gloule (Fig. 4 ) Menwhile, fish fed the floting rtificil diet (T 3 ) showed norml ovry with mture follicles, norml thecl lyer nd high content of yolk gloule (Fig. 5 ). The liver The histologicl exmintion of the liver of femle Se ss (D. lrx) rood stock fed different dietry tretments ws done too nd showed in Figs Where, fish fed the trsh fish (T 1 ) showed derrngement heptocytes, the heptocyte lining is simple lyered, low glycogen content, s well s necrosis in heptocytes ws detected (Fig. 6 c). Fish fed the mixed rtificil diet nd trsh fish (50% : 50%) (T 2 ) showed slightly derrngement heptocytes, the heptocyte lining is simple lyered, norml sinusoids, slight low glycogen content, esides the slight necrosis in heptocytes (Fig. 7 ). Menwhile, fish fed the floting rtificil diet (T 3 ) showed norml heptocytes, the heptocyte lining is simple lyered, norml sinusoids nd improvement of glycogen content in the heptocytes (Fig. 8 ). 32

7 M gi th Fig. 3 : Photomicrogrph of ovry of Se ss rood stock fed the trsh fish (T 1 ) showing ovry with degenerted mture (M) oocytes follicles nd ruptured thecl lyer (th), very low content of yolk gloules (gl), stined with & E ( 250). gi M N th Fig. 4 : Photomicrogrph of ovry of Se ss rood stock fed mixed rtificil diet nd trsh fish (50% : 50%) (T 2 ) showing ovry with irregulr mture (M) follicles, demturtion thecl lyer (th) nd fir content of yolk gloule (gl), stined with & E (, 400, nd, 250). 33

8 M gl th Fig. 5 : Photomicrogrph of ovry of Se ss rood stock fed floting rtificil diet (T 3 ) showing norml ovry with mture (M) follicles, norml thecl lyer (th) nd high content of yolk gloule (gl), stined with & E (, 250 nd, 400). G N N Fig. 6 c: Photomicrogrph of liver of Se ss rood stock fed trsh fish (T 1 ) showing derrngement heptocytes (), the heptocyte lining is simple lyered, low glycogen content (G) nd necrosis in heptocytes (N), stined with & E ( 400) c 34

9 N N S G N S Fig. 7 : Photomicrogrph of liver of Se ss rood stock fed mixed rtificil diet nd trsh fish (50% : 50%) (T 2 ) showing slightly derrngement heptocytes (), the heptocyte lining is simplelyered, norml sinusoids (S), slight low glycogen content (G) nd slight necrosis in heptocytes (N), stined with & E (, 400 nd, 100). G S S Fig. 8 : Photomicrogrph of liver of Se ss rood stock fed floting rtificil diet (T 3 ) showing norml heptocytes (), the heptocyte lining is simple lyered, norml sinusoids (S) nd improvement of glycogen content (G), stined with & E (, 100 nd, 400). Condition fctor (K f ) nd length-weight reltionship (LWR) re two importnt prmeters for studying the growth, rte of feeding, metmorphosis, ftness, onset of mturity, gondl development nd generl well eing of the fish popultion (Arned et l., 2008). In the present study, Europen Se ss (D. lrx) rood stock femle fed the rtificil diet (T 3 ) hd improved the k f mong other dietry tretments. Where, higher k f indicted good helth with n isometric growth, which is desirle for fish in fish frms (Ayode, 2011). This superiority of the rtificil diet my e relted with its high qulity components nd nutrients composition (Tles 2 nd 3). Although mles use less energy for gond mturtion thn femles (Jonsson et l., 1991), s well s it is plusile tht the mount of surplus energy ville 35

10 (from different diet) to improve their K f is higher. Moreover, mles generlly utilize less energy for gond mturtion thn femles (Jonsson et l., 1991), s well s mles of Europen se ss grow 20 40% less thn femles (Mylons et l., 2003). In the present study, no significnt differences on the mle s K f were found mong dietry tretments (Tle 4). Furthermore, mle gondl tissue does not occupy the ody cvity to the sme extent s tht of femle during lter stges of nnul reproductive development. Thus, there is likely to e more spce ville for food within mle compred to femle s ody (Fordhm nd Trippel, 1999). From other side, Pwson nd Pickett (1996) stted tht prediction of the initil occurrence of spwning ws etter correlted to the totl length (TL) rther thn to K f nd speculted tht spwning could only tke plce once the Se ss femles hd ttined TL in excess of 42cm. Certinly in other fish species such s Atlntic slmon nd yellowtil, the condition fctor of the fish determines the occurrence of spwning nd the fecundity (Venktrmni et l., 1995; Kdri et l., 1996). Inversely with the otined results herein, Bisws et l. (2011) reported tht the trsh fish feeding system could help to produce Asin Se ss with higher growth, survivl nd optimum LWR, s well s K f, s this system etter meets up the feed requirement of the growing fish s compred to live prey feeding system. Also, El- mmdy et l. (2014) reported tht the comintion of dry pellet nd trsh fish hve improving in feed efficiency nd mximized growth rtes in D. lrx, suggesting tht mixed diets my etter cover the nutritionl requirements of D. lrx. These differences could e ttriuted to the species-specific growth rtes, losses of feed mteril during feeding the trsh fish (Shpwi et l., 2011). Also, Sim et l. (2005) reported tht s much s 30 to 50% of the trsh fish fed to fish cn e lost during feeding process. Quntittive spects in fishes re importnt tools for the study of iologicl fundmentls such s viscerosomtic, heptosomtic nd gondosomtic indices, ecuse mesurement nd nlysis these indices re very importnt in ssessing food vlue (Bolger nd Connolly, 1989). In the present study, Se ss femle fed the rtificil diet (T 3 ) incresed (p 0.05) GSI compred to fish fed the other dietry tretments (T 1 nd T 2, Tle 4). While, no significnt (p 0.05) differences of GSI mong ll dietry tretments of Se ss mle. Similrly with the otined trend herein, Adelhmid et l. (2010) found tht Oreochromis niloticus femles hve higher vlues of GSI thn mles. Tht is in greement with EL- Syed et l. (2007), who lso reported tht Tilpi zillii femles showed higher GSI vlues thn mles during the period from June to Septemer with pek in July. The positive effect of the rtificil diet (T 3 ) on GSI of Se ss femle in the current findings s positive correltion of the superiority effects of this tretment (T 3 ) on K f mong ll dietry tretments. Generlly, determintion of the reproductive mturity using GSI only is not enough, ecuse the structures within the ovry, such s oocytes t different stges, interstitil tissue with ccumultion of yolk mterils, cnnot e interpreted y weight. Direct oservtion of histologicl rchitecture is the most ccurte method to let us know exctly the stge of mturtion t which the ovry is undergoing. In cse of Se ss mle fed the trsh fish (T 4 ) incresed (p 0.05) SI compred to fish fed the other dietry tretments (T 5 nd T 6 ), ut in cse of Se ss femle no significnt (p 0.05) differences of SI mong ll dietry tretments (Tle 4). SI-vlues oserved in the present study re within the rnge of vlues previously reported for Europen Se ss (Tildi et l., 1991; Dis et l., 1998). SI higher thn 2 ws noted y Metiller et l. (1981) in Se ss fed diets contining more thn 14% ft, nd y Spyridkis et l. (1986) using diets with strch level higher thn 16%. owever, Seung-Cheol et l. (2007) reported tht no significnt differences were detected of SI vlues of red Se rem, Pgrus mjor fed mixture of four supplemented medicine hers compred to the control group. In rinow trout, it hs een oserved tht feeding high levels of digestile crohydrte increses liver size nd glycogen content (Pieper nd Pfeffer, 1979; ilton nd Atkinson, 1982). Gondl mturtion of Europen Se ss rood stock cn e rrested y reduced feeding rtions (Chtzifotis et l., 2011). Nutrition of rood stock hs profound effect on gondl mturtion nd fecundity (Wtne, 1988), nd plys mjor role in reproductive success in oth fish nd crustcens (Tkeuchi et l., 1981; Teshim nd Knzw, 1983; Wtne et l., 1984). istologicl exmintion is the most ccurte tool to determine the reproductive stte of femle fish (West, 1990). In the present study, the ntomicl development of femle nd mle Se ss ws detected. Where, dietry tretments (Fig. 1, c, nd d) reveled shrply development of the ovries or the testes thn of fish pre tretments (Fig. 1 ). Prtilly, femle or mle fed the rtificil diet (T 3 nd T 6, respectively) showed more development of the gonds thn fish fed other dietry tretments. These seriously development of fish gonds were confirmed y the histologicl exmintion of the gonds nd the liver too. 36

11 These superiority of the rtificil diet regrding the ntomicl nd histologicl ltertions of the fish gonds nd the liver re not surprising. Where, these helpful effects on ntomicl nd histologicl exmintions were confirmed y the positive effects on K f, GSI nd SI prmeters (Tle 4). Where, morphologicl, nd iologicl chrcteristics of the resulting progeny re direct or indirect consequences of the previous nutrition of the rood fish during some time efore the reproductive process cn tke plce. Consequently, if the previous nutrition of rood fish is dequte, the resulting eggs cn e of good qulity nd will hve ll the potentil to produce vile fry (Crrillo et l., 2000). During oogenesis, oocytes re divided into vrious stges depending on the morphology of the nucleus, cytoplsm nd follicle. These stges my e grouped into the previtellogenic, vitellogenic, mturtion, nd tresi phses. Detils of ech stge re completely documented in Selmn nd Wllce (1989). Liver plys key role in regulting intermediry metolism in response to nutritionl sttus (Klover nd Mooney, 2004). Oservtions on the histologicl ppernce of the liver tissue herein indicted tht no pthologicl phenomen were present in Se ss femle fed the rtificil diet. Where this tretment (T 3 ) reveled tht norml heptocytes, the heptocyte lining is simple lyered, norml sinusoids, norml pncretic cini nd improvement of high glycogen content in the heptocytes (Fig. 8 ). Inversely with the current findings, mny uthors hve shown tht rtificil diets such s dry grnules induce heptic ltertions, with excessive ft in the heptocytes (Acuigrup, 1981; Strch et l., 1983). The unusully ple (nerly white) livers sometimes hd mottled ppernce indictive of ftty degenertion. Generlly, the histologicl exmintion of the ovry or the liver of Se ss rood stock fed the experimentl diets reveled the positive ltertions, especilly for fish fed the floting rtificil diet (T 3 ) thn those fed the trsh fish (srdine nd clmri, T 1 ), or fed the mixed rtificil diet nd trsh fish (50% : 50%, T 2 ). These positively effects of dietry tretments on histologicl ltertions of the ovry or the liver were potentilly relted with the ntomicl modifictions (Fig. 1), or K f, GSI nd SI (Tle 4). From other side, in the present study Se ss femle fed other dietry tretments showed some histopthologic findings of the liver (Figs. 6 nd 7) such s, derrngement heptocytes, the heptocyte lining is simple lyered, low glycogen content, necrosis in the heptocytes. In these dietry tretments elevtion is chrcteristic of liver dmge, were well correlted with the histopthologic findings of the liver. These impired ltertions in the liver tissue were mnifested s n ccumultion of lipid gloules nd reduction in glycogen reserves (Bt et l., 1983; Strch et l., 1983). This excess ft ws correlted with ltertion of the lipid metolism, secondry to deficiency in essentil ftty cids (Roche et l., 1982, ; Bt et l., 1983). Moreover, lmost every ttempt to completely replce fresh food with rtificil diets, results in decrese in ovrin mturtion, reduced numer of spwns nd n inferior egg qulity. In most situtions, comintion of fresh food nd rtificil diets gives etter results thn feeding regime tht consist of fresh food only (Bry et l., 1990, ; Nscimento et l., 1991). Conclusion The otined findings herein reveled tht fish fed the rtificil diet positively ffects on K f, GSI nd SI, esides its helpful effects on ntomicl nd histologicl exmintions of the ovry nd the liver mong other dietry tretments. ence, it could e confirmed on the potentil reltionship etween the fish feeding sources nd the reproductive efficiency of Se ss D. lrx rood stock. Bsed on this positive reltionship, the good qulity rtificil diets must e used in mrine fish htcheries to enhnce their reproductive performnce. Also, more dvnced reserches could e conducting to investigte the effect of nturl or synthetics feed dditives on the reproductive performnce of mrine fish. Conflict of interest sttement Authors declre tht they hve no conflict of interest. References Adelhmid, A.M., Mehrim, A.I., El-Brry, M.I., El- Shrwy, M.A., An ttempt to improve the reproductive efficiency of Nile tilpi rood stock fish. Fish Physiol. Biochem. 36(4), Acuigrup, J., Aspects of fish pthology in Spin, with ppliction to other geogrphic res. World Conference on Aquculture, Venice, Septemer Arned, C., Neir, R., Lm, N., Iturr, P., Slmonids. In: Genome Mpping nd Genomics in Fishes nd Aqutic Animls (Eds.: Kocher, T., Kole, C.). Springer Verlg, Berlin. pp Ayode, A. A., Length weight reltionship nd diet of Africn Crp Leo ogunensis (Boulenger, 1910) in 37

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