Badawi M. El-Sayed, Y. Hakim, Hassanin M. El-Sayed and Haytham A. Ali

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2 World Journl of Fish nd Mrine Sienes 6 (2): , 2014 ISSN IDOSI Pulitions, 2014 DOI: /idosi.wjfms Effet of Prtil Replement of Yellow Corn y Pomegrnte Peel with or Without Allzyme SSF on Growth Performne nd Helth Sttus of Oreohromis nilotius Bdwi M. El-Syed, Y. Hkim, Hssnin M. El-Syed nd Hythm A. Ali 1Deprtment of Nutrition nd Clinil Nutrition, Fulty of Veterinry Mediine, Zgzig University, Egypt 2Deprtment of Fish Diseses nd Mngement, Fulty of Veterinry Mediine, Zgzig University, Egypt 3Deprtment of Biohemistry, Fulty of Veterinry Mediine, Zgzig University, Egypt Astrt: The present work ws rried out to study the effet of prtil replement of yellow orn y pomegrnte peel (s unonventionl feedstuff) with nd without ddition of multienzymti omplex Allzyme SSF (Solid stte fermenttion) in the diets of Oreohromis nilotius nd its effet on growth performne nd helth sttus. A totl of 300 Oreohromis nilotius with n verge ody weight of 25.00±1.00 g were used. The fish were rndomly divided into 5 equl triplite groups (Eh replite ontined 20 fish). A sl ontrol diet ws formulted to fulfil the nutrients requirement of fish ording to NRC nd ontined 30.80% CP nd 2940 kl/kg DE. In other 4 experimentl diets yellow orn ws prtilly repled y pomegrnte peel t rte of 5 or 10% with nd without ddition of Allzyme SSF t rte of 0.02%. The fish were fed isonitrogenous, isolori diets 4 times dily t rte of 5% of ody weight for 10 weeks. The otined results reveled tht the fish fed diet ontined 5% pomegrnte peel with Allzyme hd higher finl ody weight, ody gin nd ody gin % s ompred to fish fed other dietry tretments. Some immunologil prmeters (IgM nd lysozyme) were signifintly improved due to supplementtion of the pomegrnte diets with Allzyme. There were no signifint vrition in different iohemil lood prmeters (ALT, AST, retinine nd ortisol) etween the ontrol group nd other groups while the generl ntioxidnt sttus ws improved due to supplementtion of pomegrnte diets espeilly with enzyme with improvement of generl helth ondition, ondition ftor nd survivility in ll experimentl groups. It ould e onluded tht pomegrnte peel ould reple up to 10% of yellow orn in Oreohromis nilotius diets with ddition of Allzyme SSF without ny dverse effets on growth prmeters or helth sttus. Key words: Oreohromis nilotius Pomegrnte Peel Allzyme Growth Immunity INTRODUCTION tnnins, suh s punilin, punilgin, pedunulgin nd punigluonin [3]; it hs een used in trditionl mediine There is growing interest within the feed for treting dirrhe, dysentery, stomhhe nd wound industry in Egypt to use unonventionl less expensive heling [4, 5]. In this regrd, stringeny is known gro-industril y-produts for lest ost effetive phrmologil property of tnnins [5, 6] nd tnnin-rih formultion nd lso to produe high qulity produts peel y-produts hve een extensively used in niml to stisfy ustomer needs in ost effetive mnner. feeds y developed ountries suh s the USA [7]. It is The pomegrnte, Puni grntum, is n edile fruit rih reognized for mny helth-promoting qulities nd soure of ntioxidnt polyphenols, minly hydrolyzle pprent wound-heling properties [8], ntimiroil tnnins nd nthoynins [1]. tivity [4], ntiner property [9], ntitherosleroti, The dried peel (Perirp) of pomegrnte fruit is ntioxidtive pities [10] nd ntioesity [11]. hrterized y their low energy density nd high Fish fed with pomegrnte enrihed diet hd ontent of mironutrients, fier nd other omponents improved hemtologil nd iohemil profiles with funtionl properties suh s phytohemils [2]. nd innte immune response ginst P. dientrrhi It ontins signifintly higher levels of hydrolysle [12]. Corresponding Author: Hythm A. Ali, Deprtment of Biohemistry, Fulty of Vet. Med., Zgzig University, Egypt. 182

3 World J. Fish & Mrine Si., 6 (2): , 2014 A wy to inrese the nutritive vlue of MATERIALS AND METHODS gro-industril y-produts of pomegrnte peel in fish diets y inlusion of enzymes tht n e used s Experimentl Fish: A totl numer of 300 pprently lterntive to inrese the feed effiieny in the rered helthy live Oreohromis nilotius with n verge ody speies nd improve the utiliztion of nutrients oming weight 25.00±1.00 g nd verge ody length 10.00±2.00 from less ost plnt ingredients tht susequently m were otined from Ass Fish Hthery t Shrki reduing prodution osts y inresing digestiility, provine. Fish were kept in glss quri ( m) it redues the lod of pollutnts from wste nd filled with 90 L, de-hlorinted fresh wter, supplied with enhnement of the produtive indies [13, 14]. ertor nd thermosttilly ontrolled t 22±2 C. The present study ws imed to evlute the effet of Fish were divided into 5 groups; eh group ws prtil replement of yellow orn y pomegrnte peel s divided into sugroups. Eh sugroup hs three unonventionl Allzyme SSF feedstuff with nd without replites (20 fish replite). The fish were dpted to ddition of enzyme omplex on growth performne nd experimentl ondition for two weeks efore the strt of helth sttus in the diets of Oreohromis nilotius. the experiment. Tle 1: Chemil nlysis of feed stuffs used in the experimentl diet Nutrient (% s fed sis) Ingredient DM CP EE CF Ash NFE(lulted) Yellow orn Pomegrnte peel Whet flour Soyen mel Fish mel Poultry y-produt mel (DM= Dry mtter, CP= Crude protein, EE= Ether extrt, CF= Crude fier nd NFE= Nitrogen free extrt). Tle 2: Ingredients nd lulted omposition of the experimentl diets Experimentl diet % Pomegrnte Peel 10 % Pomegrnte Peel Ingredients (%) Control Without enzyme With enzyme Without enzyme With enzyme Yellow orn Pomegrnte peel Whet flour Soyen mel, 44% Fish mel, 60% Poultry y-produt mel Vegetle oil Vitmin nd minerl mixture* Allzyme SSF Clulted omposition DM % CP % EE % CF % Ash % NFE % DE, Kl/ kg diet** * Vitmin nd Minerl mixture (Alfkem):-Eh 1 kg ontins:-vit. A I.U, Vit. D I.U, Vit. E. 720 mg, Vit. K3 142 mg, Vit C 0.1 mg, Vit B1 58 mg, Vit B2 34 mg, Vit. B6 34 mg, Vit.B12 58 mg, Foli id 86 mg, Pntotheni id 8 mg, Mngnese sulfte 65 mg, Zin methionine 3000 mg, Iron sulfte 2000 mg, Copper sulfte 3400 mg, Colt sulfte 572 mg, Sodium selenite 25 mg, Clium iodide 25 mg, Clium ronte (Crrier sustne) till 1000 g. ** digestile energy lultion sed on vlues of protein 3.5 kl/gm, ft 8.1 kl/gm, NFE 2.5 kl/g Sntigo et l. [17]. 183

4 World J. Fish & Mrine Si., 6 (2): , 2014 Fish Diets nd Feeding: Diets were prepred t Fish Immunogloulin M (IgM) Determintion: Reserh Unit, Fulty of Veterinry Mediine, Immunogloulin M (IgM) ws determined using Zgzig University. Feedstuffs used in diets formultion ELISA Kit. Ctlog No. CSB-E12045Fh (96k test). were nlyzed for moisture, rude protein, ether extrt CUSABIO BIOTECH CO., Ltd. nd rude fier ording to the stndrd proedures [15] nd the dt re shown in Tle 1. Diets in the form of dry Lysozyme Determintion: The lysozyme tivity ws pellets nd were formulted to meet the nutrient mesured using the turidity ssy. Chiken egg requirements of Nile tilpi s set y NRC [16] s shown lysozyme (Sigm) ws used s stndrd nd 0.2 mg ml 1 in Tle 2. The dietry tretments inluded: (1) sl lyophilized Miroous lysodeiktius in 0.04 M sodium ontrol diet ws formulted to fulfil the nutrients phosphte uffer (ph 5.75) ws used s sustrte. requirement of fish ording to NRC nd ontined 1 Fifty ml of serum ws dded to 2 ml of the teril 30.80% CP nd 2940 kl/kg DE. In other 4 experimentl suspension nd the redution in the sorne t 540 nm diets yellow orn ws repled y pomegrnte peel t ws determined fter 0.5 nd 4.5 min inution t 22 C. rte of 5 or 10% with nd without ddition of enzyme One unit of lysozyme tivity ws defined s redution omplex Allzyme SSF t rte of 0.02%. The fish were fed in sorne of 0.001min 1 [22]. diets 4 times dily t rte of 5% of ody weight for 10 weeks. Alnine Aminotrnsferse Ativity (ALT) nd Asprtte Aminotrnsferse (AST) Determintion: ALT nd AST The Allzyme SSF: The Allzyme SSF ws used s n ws determined (By Spetrum Kits, Egyptin Compny for enzyme omplex; it ontins unique formul of Biotehnology, Ciro, Egypt (REF: for ALT nd fermenttion produts from Aspergillus niger. Eh one for AST) following the steps reommended y kits g ontins phytse (300 SPU), protese (700 HUT), ording to Reitmn nd Frnkel [23]. ellulose (40 CMCU), xylnse (100 XU), etglunse (200 BGU), mylse (30 FAU) nd petinse (4000 AJDM). Assy Proedure for GSH Determintion: Produed nd exported from Co., S.A. Mexio ws Redued glutthione (GSH) ws determined in the purhsed from IVT vlue vet. Egypt. liver homogente, using kit supplied y Cymn (Ct. No , Cymn, USA) ording to the Growth Performne Prmeters: The fish were mnufturer s instrutions [24]. weighted t the strt nd the end of the experiment. Averge ody weight ws lulted y dividing the totl Cretinine Determintion: It ws rried out weight of fish y the numer of fish in eh group. Body ording to method mentioned y Husdn nd Rpoport gin nd feed onversion rtio [18]. Body gin perent [25]. [19] nd speifi growth rte % [20] were determined. Cortisol Level Determintion: Serum ortisol ws Blood Smple Colletion: Two Blood smples t the determined y ELISA using Mirotiterstrip DIAMED eginning nd t the end of experimentl period were CORTISOL ELISA kit whih sed on the priniple of otined from udl lood vessels into plsti Eppendorf ompetition etween leled (Cortisol Enzyme tues for serum smples preprtion. Blood ws olleted Conjugte) nd n unleled steroid (Cortisol for inding into Eppendorf tues without ntiogulnt in syringe to Cortisol speifi ntiodies, oted into the wells of then entrifuged (3000 rpm for 15 min). The serum smples miro titer plte). were olleted nd stored immeditely in freezer (zero C) until use [21]. Condition Ftor: The ondition ftor ws lulted ording to Gjedrem nd Gunnes [26]. Liver Smple Colletion: At the eginning nd t the end of experimentl period one hlf of grm of livers were Helth Condition: For evlution of helth ondition of tken nd homogenized in 5ml old 20 mm HEPES uffer, the fish during the period of the experiment, espe, ph 7.2, ontining 1mM EGTA, 210 mm mnnitol nd 70 defensive, til nd oulr reflexes were regulr oserved mm surose per grm tissue. Homogentes were ording to Luky [27]. entrifuged t x g for15 minutes t 4 C nd the Fish of ll groups were regulr oserved dily, superntnt ws removed nd stored t-80 C until further norml ehviors nd mortlity rte were determintion of GSH. reorded. 184

5 World J. Fish & Mrine Si., 6 (2): , 2014 Sttistil Anlysis: The otined dt in this study feed intke ws deteted mong the dietry tretments. were sttistilly nlyzed for vrine ANOVA, LSD The totl FCR (In the entire experimentl period) (Lest signifint differene) ording to Snedeor nd in fish group fed on diet ontined 10% pomegrnte Cohrn [28]. Differenes mong tretment mens were peel without llzyme ws signifintly (P<0.05) ompred using Dunn's multiple rnge tests [29]. deresed, wheres other dietry tretments displyed Dt were presented s men ± SE nd signifine ws non-signifint (P>0.05) differenes in the totl delred t (P<0.05). FCR ompred to the ontrol group. However, fish group fed the diet ontined 5% pomegrnte RESULTS peel with llzyme hd utilized their feed more effiiently thn other dietry tretments or the Growth Performne: The effet of dietry tretments on ontrol. growth performne of Oreohromis nilotius is present in Tle 3. In onlusion, ody weight, BW gin, BW gin Immunologil Prmeters: There were signifint % nd speifi growth rte % weren't signifintly differenes in immunologil prmeters (P<0.05) mong (P>0.05) different mong the dietry tretments, fish groups. The fish groups fed on diet ontined 5% or wheres in fish group fed on diet ontined 10% 10% pomegrnte peel with enzyme hd signifint pomegrnte peel without llzyme ws signifintly higher vlues of finl IgM nd lysozyme vlues s (P<0.05) deresed if ompred to other dietry tretments ompred with other dietry tretments or the ontrol s or the ontrol. However, fish group fed the diet ontined shown in Tle 4. 5% pomegrnte peel with llzyme hd reorded higher Regrding to the effet of dietry supplementtion vlues; while fish group fed on diet ontined 10% with different growth promoters on different lood pomegrnte peel without llzyme hd reorded lower prmeters of O. nilotius, there were no signifint vlues for ll growth performne prmeters s ompred vrition etween the ontrol group nd other groups s to the ontrol. There wsn't signifintly (P>0.05) different shown in Tle 5. Tle 3: The effet of dietry supplementtion with different growth promoters on llover performne of O. nilotius Experimentl diet Groups 5 % Pomegrnte Peel 10 % Pomegrnte Peel Prmeter Control diet Without llzyme With llzyme Without llzyme With llzyme Initil ody weight (g) 24.95± ± ± ± ±0.20 Finl ody weight (g) ± ± ± ± ±0.45 Body weight gin (g) 7.37± ± ± ± ±0.54 Body weight gin (%) ± ± ± ± ±2.29 Speifi growth rte (%) 0.46 ± ± ± ± ±0.03 Feed onsumption (g) ± ± ± ± ±0.39 Feed onversion rtio 3.08 ± ± ± ± ±0.21 -Mens in the sme row with different supersript letters re signifintly different t (P<0.05). Tle 4: The effet of dietry supplementtion with different growth promoters on immune sttus of O. nilotius Experimentl diet Groups 5 % Pomegrnte Peel 10 % Pomegrnte Peel Prmeter Control diet Without llzyme With llzyme Without llzyme With llzyme Initil IgM vlue 17.52± ± ± ± ±1.14 Finl IgM vlue 17.70± ± ± ± ±0.67 Initil lyzozyme vlue 16.01± ± ± ± ±1.28 Finl lyzozyme vlue 16.74± ± ± ± ±0.61 -Mens in the sme row with different supersript letters re signifintly different t (P<0.05). 185

6 World J. Fish & Mrine Si., 6 (2): , 2014 Tle 5: Blood prmeters nd liver GSH fter dietry supplementtion with different growth promoters on O. nilotius Experimentl diet Groups 5 % Pomegrnte Peel 10 % Pomegrnte Peel Prmeter Control diet Without llzyme With llzyme Without llzyme With llzyme Initil ALT vlue (µg /ml) 17.21± ± ± ± ±0.16 Finl ALT vlue (µg/ml) 16.53± ± ± ± ±0.54 Initil AST vlue (µg/ml) 17.65± ± ± ± ±0.26 Finl AST vlue (µg/ml) 16.95± ± ± ± ±0.35 Initil liver GSH (mg/g tissue) 9.27± ± ± ± ±0.73 Finl liver GSH (mg/g tissue) 9.05± ± ± ± ±0.64 Initil Cretinine (mg/dl) 0.22± ± ± ± ±0.22 Finl Cretinine (mg/dl) 0.18± ± ± ± ±0.35 Initil ortisol vlue (µg/ml) 6.22± ± ± ± ±0.22 Finl ortisol vlue (µg/ml) 5.21± ± ± ± ±0.15 Tle 6: The effet of dietry supplementtion with different growth promoters on ondition ftor nd survivl rte (%) of O. nilotius Experimentl diet Groups 5 % Pomegrnte Peel 10 % Pomegrnte Peel Prmeter Control diet Without llzyme With llzyme Without llzyme With llzyme Initil ody length (m) 11.2± ± ± ± ±0.21 Finl ody length (m) 12.3± ± ± ± ±0.27 Condition ftor (k) Survivl rte (%) The results in Tle 6 showed tht oth of ondition multiple helth enefits suh s extending the lifespn of ftors nd survivl rte of the fish were idel in ll diverse orgnisms inluding mmmls [33], regultion of groups of the experiment. These results onfirm the good immune funtion [34] nd ner prevention [35]. enefiil effet of oth pomegrnte peel nd llzyme. This ws ssoited with mrked improvement in ody weight nd BW gin of fish group fed the diet DISCUSSION ontined pomegrnte peel with llzyme if ompred to those fed on the diet tht ontin the pomegrnte peel Our growth performne results for fish groups fed without llzyme whih lost weight reltive to the ontrol pomegrnte peel without enzyme greed with Mhmoud group. However, it is notied tht, repling the et l. [11] who reported tht, the rts fed on diets pomegrnte peel with llzyme used reltive ontining dry pomegrnte peel n e used sfely to improvement in growth rte of fish thn other groups mnge ody weight or even for weight redution without without llzyme. ny helth hzrds. This effet on ody weight gin or The feed intke y fish ws the sme (P>0.05) even use weight redution ould e ttriuted to etween tretments, with no leftovers oserved. As the severl ftors s pomegrnte peel ontins onsiderle sme mount of diet ws used, regrdless of tretment, mounts of polyphenols together with the high fier it ws oserved tht the tilpis otined positive ontent whih redued food onsumption nd the response from enzymti tretment, even eing restrited restrited lorie intke [11]; ontins polyphenols my to growth in its entirety. suppress growth of the dipose tissue through their The ddition of the enzyme omplex SSF in the fish ntingiogeni tivity nd y modulting dipoyte diets improves the performne (Finl weight nd weight metolism [30, 31] or ontins resvertrol, vitmin D, gin, while other performne prmeters, there were no queretin nd genistein [32]. Some reserhers hve differenes) of Nile tilpi [14] nd tht for ommon rp suggested tht mild lorie restrition might provide nd grss rp juveniles [13] fter feed supplementtion 186

7 World J. Fish & Mrine Si., 6 (2): , 2014 using lflf mel, led to the inrese of nutrients There were no signifint vrition in some pprent digestiility oeffiient nd inrese in the iohemil lood prmeters (ALT, AST, retinine nd iovilility of nutrients, thus explining the more ortisol) etween the ontrol group nd other elerted growth. groups, this men tht ddition oth of pomegrnte No effet (P>0.05) of levels of the enzyme omplex peel with or without llzyme in diet not only improve SSF ws oserved on feed onversion rtio espeilly in immunity ut lso, hve no dverse effet in fish group fed the diet ontined 5% pomegrnte peel different fish orgns nd helth sttus of fish. with llzyme. These results re in greement with Sores These results re greed with Mhmoud et l. [11] et l. [36], who reported tht the inlusion of exogenous who reported tht the fish fed on diets ontining dry protese improved feed onversion, weight gin nd pomegrnte peel n e used sfely to mnge ody speifi growth rte for Tuunre p juveniles weight or even for weight redution without ny (Cihl temensis). helth hzrds. Also in tle 5 there were inreses There ws signifint differenes (P<0.05) in some in the level of GSH in the groups treted with immunologil prmeters etween fish groups. The fish pomegrnte peel espeilly tht treted with ddition of groups fed on diet ontined 5% or 10% pomegrnte enzyme whih refers to its ntioxidnt tivities whih peel with enzyme hd signifint higher vlues of finl n improve generl helth sttus of the treted groups IgM nd lysozyme s ompred with other dietry [10] tretments or the ontrol. The ondition ftor nd the survivl rte of ll These results re in greement with Hrikrishnn experimentl groups were idel, these results were et l. [12] who reported tht the serum lysozyme tivity ompletely onfirmed with Brg et l. [4], signifintly (P<0.05) enhned from weeks 1 to 4 in fish Chidmr et l. [8], Jeune et l. [9] nd fed with pomegrnte enrihed diet nd hllenged with Tzulker et l. [10] who mentioned tht pomegrnte or without P. dientrrhi when ompred to ontrol. peel hs ntimiroil tivity, ntiner property, The innte immune response generlly preedes the ntitherosleroti, ntioxidtive pities. dptive response it tivtes nd determines the nture of the dptive response nd o-opertes in the CONCLUSION mintenne of homeostsis [37]; it is prtiulrly ttriuted to the phgoyti, lysozyme nd spontneous It ould e onluded tht fish fed with pomegrnte hemolyti tivities [38]. peel ould reple up to 10% of yellow orn in Lysozyme is n importnt prmeter in the immune Oreohromis nilotius diets with ddition of Allzyme SSF defense of oth invertertes nd vertertes. Lysozyme without ny dverse effets on growth prmeters or is teriidl tivity, hydrolyzing-linked glyoside immune sttus. onds of teril ell wll peptidoglyns result in lysis [37]. The protetive response in terms of umultive REFERENCES mortlity ws signifint (P<0.05) lower vlues in groups fed on diet ontined 5% or 10% pomegrnte peel with 1. Irhim, M.I., Effiieny of Pomegrnte Peel or without enzyme ginst infetion when ompred to Extrt s Antimiroil, Antioxidnt nd Protetive ontrol. Therefore, we suggest tht pomegrnte enrihed Agents. World Journl of Agriulturl Sienes, diet following hllenge with infetion improves the 6(4): innte immune system in Oreohromis nilotius ginst 2. Vn Duyn, M.A.S. nd E. Pivonk, infetion. These results re in greement with "Overview of the helth enefits of fruit nd Hrikrishnn et l. [12] who reported tht the umultive vegetle onsumption for the dietetis professionl: mortlity ws high, 80%, in fish fed non-pomegrnte seleted literture", J. Am. Diet Asso., enrihed diet ginst P. dientrrhi. However, fish fed 100: with pomegrnte enrihed diet nd hllenged with or 3. Dudonne, S., X. Vitr, P. Coutiere, M. Woillez nd without P. dientrrhi were low t 50% nd 10%. J.M. Merillon, Comprtive study of ntioxidnt The pomegrnte enrihed diet enhned the innte properties nd totl phenoli ontent of 30 plnt immune response, whih my possily e n importnt extrts of industril interest using DPPH, ABTS, ftor in reduing the perentge umultive mortlity FRAP, SOD nd ORAC ssys. J. Agri. Food Chem., therey proteting the fish from P. dientrrhi infetion. 57:

8 World J. Fish & Mrine Si., 6 (2): , Brg, L., J. Shupp, C. Cummings, M. Jett, 14. Mour, G.S., E.A.T. Lnn, K. Filer, D.L. Flkoski, J. Tkhshi, L. Crmo, E. Chrtone-Souz nd J. L. Donzele, M.G.A. Oliveir nd S.T. Rezende, A. Nsimento, Pomegrnte extrt inhiits Effets of enzyme omplex SSF Stphyloous ureus growth nd susequent (solid stte fermenttion) in pellet diets for Nile enterotoxin prodution. J. Ethnophrmol., tilpi. Revist Brsileir de Zooteniv, 96: (10): Reddy, M.K., S.K. Gupt, M.R. Jo, S.I. Khn nd 15. AOAC (Assoition of Offiil Anlytil Chemists), D. Ferreir, Antioxidnt, ntimlril nd Offiil methods of nlysis. Assoition of ntimiroil tivities of tnnin-rih frtions, offiil nlytil hemists. Arlington, VA, ellgitnnins nd phenoli ids from pp: Puni grntum L. Plnt Med., 73: NRC (Ntionl Reserh Counil), Prshnth, D., M. Ash nd A. Amit, Nutrient Requirements of fish. Ntionl Ademy Antiteril tivity of Puni grntum. Press, Wshington, DC, pp: 112. Fitoterpi, 72: Sntigo, C.B., M. Bnes-Ald nd M.A. Lron, 7. Al-Zoreky, N.S., Antimiroil tivity of Dietry rude protein requirement of pomegrnte (Puni grntum L.) fruit peels. Int. J. Tilpi niloti fry Kliksn, philipp. J. Biol., Food Miroiol., 134: (2-3): Chidmr, M.K., V.K. Reddy, J.M. Veigs nd 18. Siddiqui, A.Q., M.S. Howlder nd A.A. Adm, U.D. Murthy, Study on wound heling tivity Effets of dietry protein levels on growth, of Puni grntum peel. J. Med. Food, feed onversion nd protein utiliztion in fry nd 7: young Nile Tilpi (Oreohromis nilotius). 9. Jeune, M.A., J. Kumi-Dik nd J. Brown, Aquulture, 70: Antiner tivities of pomegrnte extrts nd 19. Juney, K. nd B. Ross, A guide to Tilpi genistein in humn rest ner ells. J. Med. Food, feeds nd feeding. University of Sterling, institute of 8: Aquulture, Sterling, Sotlnd. 10. Tzulker, R., I. Glzer, I. Br-Iln, D. Hollnd, 20. Pouomonge, V. nd J. Monglng, M. Avirm nd R. Amir, Antioxidnt tivity, Effet of feeding rte on the growth of tilpi polyphenol ontent nd relted ompounds in (Oreohomis niloius) in erthen ponds. Bmidegh, different fruit juies nd homogentes prepred from 45: different pomegrnte essions. J. Agri. Food 21. Aly, S.M., Y.A.G. Ahmed, A.A.A. Ghree nd Chem., 55: M.F. Mohmed, Studies on Billus sutilis 11. Mhmoud, M.H., S.S. Kssem, M.M. Adel-Kder nd Ltoillus idophilus, s potentil nd F.A. El-Shoki, How to redue weight nd proiotis, on the immune response nd resistne keep. Interntionl J. of Ademi Reserh, of Nile tilpi (Oreohromis nilotius) 3(6): to hllenge infetions. Fish Shellfish Immunol., 12. Hrikrishnn, R., J. Kim, M. Kim, C. Blsundrm 25: nd M. Heo, Pomegrnte enrihed diet 22. Prry, R.M., R.C. Chndn nd K.M. Shhni, enhnes the hemtology, innte immune response A rpid nd sensitive ssy of murmidse. nd disese resistne in olive flounder ginst Pro. So. Exp. Biol. Med., 22: Philsterides dientrrhi. Veterinry Prsitology, 23. Reitmn, S. nd S. Frnkel, A olorimetri 187: method for the determintion of glutmi-oxloeti 13. Aurel,., M. Florin, G. Erol nd B. Mihi, nd glutmi-pyruvi trnsminses. Am. J. Clin. The Influene of Multienzymti Complex Allzyme Pthol., 33: SSF on Prodution Performnes of Common Crp 24. Ellmn, G.L., Tissue sulfhydryl groups. (Cyprinus rpio L.) nd Grss Crp Arh Biohem. Biophys., 17: (Ctenophryngodon idell V.) Juveniles fter 25. Husdn, H. nd A. Rpoport, Estimtion of Introdution of Alflf Mel in Feed. Animl Siene retinine y Jffe retion. A omprison of three nd Biotehnologies, 45(2): methods. Clin. Chem., 14:

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