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Effects of diets with soybean meal on the growth, digestibility, Phosphorus and Nitrogen excretion of juvenile rainbow trout Oncorhynchus mykiss

118 Hidroiológic Cruz 2011, Cstro 21 (2): C. A et l. Effects of diets with soyen mel on the growth, digestiility, Phosphorus nd Nitrogen excretion of juvenile rinow trout Oncorhynchus mykiss Efectos
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118 Hidroiológic Cruz 2011, Cstro 21 (2): C. A et l. Effects of diets with soyen mel on the growth, digestiility, Phosphorus nd Nitrogen excretion of juvenile rinow trout Oncorhynchus mykiss Efectos de diets con hrin de soy en el crecimiento, digestiilidd, excreción de fósforo y nitrógeno de juveniles de truch rco iris Oncorhynchus mykiss Crlos Agustín Cruz Cstro, Luis Héctor Hernández Hernández, Mrio Alfredo Fernández Ariz, Teres Rmírez Pérez nd Omr Angeles López Lortorio de Producción Acuícol (Acurio), Fcultd de Estudios Superiores Iztcl. UNAM. Av. de los Brrios 1, Los Reyes Iztcl, Tllnepntl, Edo. de México, México e-mil: Cruz Cstro C. A., L. H. Hernández Hernández, M. A. Fernández Ariz, T. Rmírez Pérez nd O. Ángeles López Effects of diets with soyen mel on the growth, digestiility, Phosphorus nd Nitrogen excretion of juvenile rinow trout Oncorhynchus mykiss. Hidroiológic 21(2): ABSTRACT A feeding tril ws performed to evlute the inclusion of high levels of soyen mel s sustitute for fishmel in diets for juvenile rinow trout nd determine the phosphorus nd nitrogen excretion. Fishmel ws replced with soyen mel t levels of 50, 75 nd 100 % with n inclusion of 0.8 g of phytse per kg of diet (g kg -1 ), wheres diet with 100% sustitution ws used without phytse. A commercil diet nd one with 100% of fishmel were used s controls. Diets were fed to triplicte groups of juveniles (4.12 ± 0.7 g, men ± stndrd devition) for period of 50 dys, nd growth performnce, P nd N excretion nd protein digestiility were determined t the end of this period. Dt were nlyzed with one-wy ANOVA. The results showed tht fish growth decresed when 100% of sustitution ws used, ut diet with 75% of soyen mel did not ffect growth. In ddition, excretion vlues of P nd N t this level of sustitution were lower thn those of the fish fed the control diets. The dt suggest tht diet with 75% soyen mel nd 25% fishmel might e used under prcticl conditions, s the growth performnce oserved on the juveniles fed such diet ws similr to those of fish fed the commercil diet nd the nutrient loding ws reduced. Key words: Rinow trout, Oncorhyncus mykiss, soyen mel, fishmel, sustitution, phytse. RESUMEN Se relizó un prue de limentción pr evlur l inclusión de ltos niveles de hrin de soy como sustituto de l hrin de pescdo en diets pr juveniles de truch rco iris y determinr l excreción de fósforo y nitrógeno. L hrin de pescdo se sustituyó en niveles de 50, 75 y 100% con l inclusión de 0.8 g de fits por kg de diet (g kg -1 ), sí como un diet con 100% de sustitución sin fits fue utilizd. Un diet comercil y un con 100% de hrin de pescdo se utilizron como controles. Ls diets se ofrecieron grupos de juveniles por triplicdo de juveniles (4.12 ± 0.7 g, promedio ± desvición estándr) por un periodo de 50 dís. Al finlizr este periodo, se determinó el crecimiento, l excreción de fósforo y nitrógeno y l digestiilidd de l proteín. Los dtos se nlizron con un ANDEVA de un ví. Los resultdos mostrron que el crecimiento de los peces disminuyó cundo se utilizó 100% de sustitución con o sin fits, pero el 75% de hrin de soy en l diet no fectó el crecimiento. A este nivel de sustitución tmién se oservron vlores de excreción de fósforo y nitrógeno más jos que los encontrdos en los peces limentdos con ls diets control. Los dtos sugieren que l diet con 75% de hrin de soy y 25% de hrin de pescdo puede ser uti- Hidroiológic Soyen mel in diets of rinow trout 119 lizd en condiciones práctics de cultivo, pues el crecimiento oservdo en los juveniles limentdos con dich diet fue similr l de los peces limentdos con l diet comercil y demás, se redujo el porte de nutrientes l gu. Plrs clve: Truch rcoiris, Oncorhyncus mykiss, hrin de soy, hrin de pescdo, sustitución, fits. INTRODUCTION The rpidly expnsion of quculture during the lst 20 yers hs een ccompnied y n incresing demnd for qufeeds (Gtlin et l., 2007). Fishmel is the mjor source of protein in most feeds (Drew et l., 2007), ut it is ecoming n expensive resource ecuse of its rising demnd (Dlsgrd et l., 2009). Moreover, the use of fishmel for qufeeds hs negtive effects on the environment, since its use increses the level of nutrients s phosphorus (P) nd nitrogen (N) in wstewters from the frms (Hrdy, 2002). The use of plnt mels hs een suggested s n lterntive to fishmel: they re cheper, redily ville nd their phosphorus content is lower thn tht of fishmel (Dlsgrd et l., 2009). Among plnt mels, soyen mel hs een pointed out s the most promising lterntive sources of protein source for qufeeds (Hrdy, 1996) ecuse of its protein level (44-48%), resonle well-lnced mino cid profile, vilility nd cost (Cheng et l., 2004; Gtlin et l., 2007). The use of soyen mel s prtil sustitute of fishmel hs een reported for severl species of fish (Fowler, 1980; Olli et l.,1989; Arnesen et l., 1989; Krogdhl, 1994; Arndt et l., 1999; Refstie et l., 2000; Refstie et l., 2001). However, its use is still limited (up to 25% of sustitution), in prt ecuse of the lrge quntity of indigestile crohydrtes, lthough the presence of phytic cid or myo-inositol hexphosphte is the mjor concern (Lining et l., 2010). Phytic cid is the min form of storing P in seeds (Jorquer et l., 2008), nd usully cuses low vilility of minerls nd reduces the pprent digestiility of protein (Gtlin et l., 2007). The most efficient wy to reduce the effect of this cid is y using phytse, phosphohydrolse enzyme tht ctlyzes the sequentil relese of inorgnic orthophosphte from phytic cid (Jorquer et l., 2008) nd hs the potentil to improve digestiility of nutrients for qutic orgnisms (Cheng & Hrdy, 2003). The use of soyen nd phytse in rinow trout (Oncorhyncus mykiss Wlum, 1792) hs een reported y Cheng nd Hrdy (2003), who found tht inclusion of phytse incresed the pprent digestiility coefficients of crude protein nd minerls of diet prepred with full-ft soyen mel. Cheng et l. (2004) reported tht the pprent digestiility of crude protein of diets prepred with soyen mel in rinow trout juveniles incresed using microil phytse. The sme uthors suggested 500 FTU of phytse kg -1 diet s optiml for the rinow trout. Wng et l. (2009) reported tht use of soyen mel pretreted with phytse or spryed phytse llowed improvement of the pprent digestiility coefficient of diets, nd tht pretretment t level of 1,000 U of phytse/kg diet ws the est option. In spite of those results, there is no report of how high levels of inclusion of soyen mel might ffect the levels of dissolved nitrogen (N) nd phosphorus (P) excretion, which is mjor concern for mny fish culture opertions (Bureu & Cho, 1999). Thus, the im of the present reserch ws to evlute the inclusion of high levels of soyen mel s sustitute of fishmel in diets for juvenile rinow trout, nd to determine the P nd N excretion. MATERIAL AND METHODS Test fish nd test diets. Rinow trout, Oncorhynchus mykiss, of n ge of 60 dys post-htching, were otined from privte frm in the Stte of Mexico, Mexico nd trnsported to the Lortorio de Producción Acuícol, Fcultd de Estudios Superiores Iztcl, Universidd Ncionl Autónom de México. Fish were mintined in 500-L tnks nd fed commercil pelleted diet for trout (Api-truch 1, 50% protein, Mlt-Cleyton de México, México) until 7 d efore the feeding tril. Four test diets were formulted with different levels of sustitution of soyen mel (Protein, 50.6 ± 1 %; totl phosphorus, 0.25 ± %; Pronsoy, S.A. de C.V., México, México) for fishmel. The formultion of these diets re shown in Tle 1 nd re sed on the species requirements reported y NRC (1993), with minimum of 40% of protein content. Cod liver oil (Drots, S.A. de C.V., México, México) nd soyen lecithin (Astecedor de Productos Nturles, S.A. de C.V., Mérid, Mexico) were used s dditionl lipid sources, wheres dextrin (Droguerí Cosmopolit S.A. de C.V., Mexico, Mexico) ws used s crohydrte source. Moreover, vitmin nd minerl mix (Micro Rovimix for crnivorous fish, (DSM Nutritionl Products de Mexico, S.A. de C.V., Gudljr, Mexico) nd whet gluten s inder (Sigm Aldrich Co., St. Louis, MO, USA), were used. α-cellulose (Sigm Aldrich Co., St. Louis, MO, USA) ws dded s filler up to 100%. Three of the four experimentl diets (sustitution of 50, 75 nd 100%, Soy1, Soy2 nd Soy3) included 0.8 g of phytse (Phytse Ronozyme P5000, DSM Nutritionl Products de México, S.A. de C.V., Gudljr, México) per kg of diet (g kg -1 ). The fourth diet (Soy4) ws with 100% sustitution, nd no enzyme dded. A diet with 100% of fishmel (Tle 1) nd commercil diet (Api-truch 1, 50% protein, Mlt-Cleyton de Mexico, Mexico) were used s control diets. Diets were prepred ccording to Hernández et l. (2004): riefly, the powdered ingredients were mixed with the oils nd distilled wter (40%). The wet dough ws pssed through met chopper to otin 5-mm dimeter pellets, which were dried t 60 Vol. 21 No 120 Cruz Cstro C. A. et l. Tle 1. Formultion of the test diet fed to juveniles of rinow trout. Ingredient (g/kg) Soy 1 (50%-50%) Soy 2 (25%-75%) Diets (% FM-% SM) Soy 3 (0%-100%) Soy 4 (0%-100%) Control (100%-0%) Fish mel (FM) Soyen mel (SM) Cod liver oil Soyen lecithin Dextrin Vitmin nd minerl mix Gluten Cellulose Phytse Proximte composition (%) Protein Lipid Ash Moisture Vitmin nd minerl mix (g kg -1 ): ρ-minoenzoic cid, 1.45; iotin, 0.02; myo-inositol, 14.5; nicotinic cid, 2.9; C-pntothente, 1.0; pyridoxine-hcl, 0.17; rioflvin, 0.73; thimine-hcl, 0.22; mendione, 0.17; α-tocopherol, 1.45; cynocolmine, ; clciferol, 0.03; L-scoryl-2-phosphte-Mg, 0.25; folic cid, 0.05; choline chloride, 29.65; retinol, 0.015; NCl, 1.838; MgSO 4 7H 2 O, 6.85; NH 2 PO 4 2H 2 O, 4.36; KH 2 PO 4, 11.99; C(H 2 PO 4 ) 2 2H 2 O, 6.79; Fe-citrte, 1.48; C-lctte, 16.35; AlCl 3 6H 2 O, 0.009; ZnSO 4 7H 2 O, 0.17; CuCl 2, ; MnSO 4 4H 2 O, 0.04; KI, nd CoCl 2, C in constnt temperture oven for 60 min nd stored t -20 C until used. Feeding tril. For the feeding experiment, recircultion system with 18 plstic tnks (100-L cpcity, filled with 80-L wter) ws used. Fifteen juveniles with n initil weight of 4.12 ± 0.5 g (men ± stndrd devition) were rndomly stocked in ech tnk. Ech test diet ws fed to triplicte groups of juveniles. Fish were fed ech dy the respective diet t 7% of the ody weight nd the dily rtion ws divided into two equl feedings t 800 nd 1500 h. The juveniles were weighed every 10 dys nd the rtion size ws djusted ccordingly. Ded fish were not replced during the experiment. Thirty minutes fter feeding, the remining feed ws removed nd quntified for mesurement of the diet intke nd then, the fecl mtter ws tken out y siphoning the ottom of ech tnk. Throughout the feeding tril, men (± SD) wter temperture ws14 ± 1 C, nd dissolved oxygen nd ph were 6.8 ± 1.0 mg L -1 nd 7.9 ± 0.5, respectively. The wter flow through ech tnk ws 1.5 L min -1 during the entire experiment. All tnks were mintined under nturl photoperiod (11 hours light, 13 hours drk). The feeding tril lsted 50 d, from Decemer 2007 to Jnury At the end of the 50 d, fish were strved for 24 h, nd weighed to otin growth performnce. Five fish, collected rndomly, were returned to the tnks nd were fed their respective diets dded with 1% of chromium oxide during 10 d, for pprent digestiility determintion. The remining fish were used to evlute oxygen consumption, nd P nd N excretion. Digestiility determintion. The pprent digestiility coefficients (ADC) of protein were determined with the NRC (1993) technique, using chromic oxide t concentrtion of 1% (Cr 2 O 3, powder, J.T. Bker, Phillipsurg, N.J., USA) s n inert mrker. As fish rpidly ccepted the diets with chromic oxide, no conditioning period ws deemed necessry nd the juvenile were fed with these diets for 10 dys, during which feces were collected on dily sis. Feces were dried in n oven nd digested in mixture of perchloric cid nd nitric cid t 250 C. Chromic oxide ws quntified with the eqution: CR Y Where: Y = sornce t 350 nm (Furukw & Tsukhr, 1966). The ADC were clculte with the eqution: ADC(%) = 100 %MF %PF %MH * %PH * 100 Hidroiológic Soyen mel in diets of rinow trout 121 Where: MF nd MH = mrker in feed nd in feces; PF nd PH = nutrient (protein) in feed nd in feces. Oxygen consumption, P nd N excretion. Fish were fed their respective diets 24 hours efore the test. A closed recircultion system of twenty 1-L flsks connected in series, y plstic tues ws slowly filled with wter nd one fish ws plced per flsk. Wter smples were tken (initil P nd N concentrtions = 0 mg L -1 ) nd the dissolved oxygen ws mesured, then flsks were hermeticlly closed. After 30 min, the flsks were opened, dissolved oxygen ws mesured with n oxygen meter (model 85, YSI Incorported, OH, USA), nd wter smples were tken to determine P nd mmoni N concentrtions with the molydovnte nd Nessler methods (Clesceri et l., 1998). Oxygen consumption ws clculted s the difference etween the initil nd finl concentrtion. Sttisticl nlysis. Dt on finl ody weight (FBW), weight gin (WG), specific growth rte (SGR), feed conversion efficiency (FCE), feed intke (FI), survivl, oxygen consumption, P nd N excretion nd ADC were tested for normlity nd homoscedsticity with the Shpiro nd Wilk W test nd Brlett s test, respectively (Zr, 1999), fter rcsine trnsformtion in the cse of survivl, expressed s percentge. Since ll dt were norml nd homoscedstic, the dt were compred with one-wy ANOVA tests. Significnt differences mong tretments were determined y Tukey multiple comprison tests (Zr, 1999), with significnce level of 5% (p 0.05). All sttisticl nlysis were performed with the pckge Minit version 15 (Minit Inc., Stte College PA, USA). RESULTS The mens of finl ody weight (FBW), weight gin (WG), specific growth rte (SGR), feed conversion efficiency (FCE), protein conversion efficiency (PCE) nd feed intke (FI) of the juvenile rinow trout re given in Tle 2. The fish fed with diet Soy2 showed the highest vlues of FBW, WG, FCE nd PCE mong the diets with soyen mel, nd did not show significnt differences when compred with those oserved with the control nd commercil diets. The growth performnce of the fish fed with diet Soy4, which hd 100% soyen mel without phytse, ws lower thn those of the juveniles fed with phytse-dded diets. Feed intke ws similr with diets Soy1, Soy2, control nd commercil, nd these were significntly higher thn with Soy3 nd Soy4. Survivl rnged from 66 ± 11% with diet Soy3 to % with the other diets, nd there were no significnt differences mong tretments (Tle 2). Oxygen consumption ws significntly lower in the juveniles fed diets Soy3 nd Soy4 thn those determined for the rest of the experimentl groups. The significntly highest consumption ws oserved in the fish fed the control diet with 100% of fishmel s protein source (Fig. 1). There ws tendency of lower vlues of P excretion (s mg of PO 4-3 L -1 ) with incresing soyen mel content in the experimentl diets with phytse. The excretion of the fish fed diet Soy4 (100% soyen mel without phytse) ws significntly higher thn tht oserved in the diet Soy3 (100% soyen with phytse), nd the highest vlue ws oserved in the fish fed the diet control with 100% of fishmel (Fig. 1). Tle 2. Finl ody weight (BW), weight gin (WG), specific growth rte (SGR), feed conversion efficiency (FCE) nd feed intke (FI) of juveniles of rinow trout fed diets with high levels of soyen mel Vlues re mens of the three replicte groups ± stndrd devition. Mens with different letters in the sme line differ significntly (p 0.05). Tretments Soy1 Soy2 Soy3 Soy4 Control Comm FBW (g) 10.6 ± ± ± ± ± ± 1.9 WG 1 (%) 157 ± ± ± ± ± ± 49 SGR 2 (% dy -1 ) 1.7 ± ± ± ± ± ± 0.2 FCE ± ± ± ± ± 0.07c 1.2 ±0.05c PCE ± ± ± ± ± 0.1 c 1.8 ± 0.1c FI (g fish -1 dy -1 ) 2.2 ± ± ± ± ± ± 0.1 Survivl (%) 91 ± 4 89 ± 8 66 ± ± ± 4 89 ± 8 1 Weight Gin = ( Finl BW Initil BW ) * Weight gin (g) Feed conversion efficiency = Initil BW Totl feed intke in dry weight sis 2 Specific growth rte = (ln Finl BW - ln Initil BW ) * Weight gin (g) Protein conversion efficiency = 50 Totl protein intke in dry weight sis Vol. 21 No 122 Cruz Cstro C. A. et l. NH O 2 consumption (mg fish -1 h N excretion (mg L -1 ) ) Soy1 Soy2 Soy3 Soy4 Control Comm Tretments () (c) Soy1 Soy2 Soy3 Soy4 Control Comm Tretments c PO 2 3- excretion (mg L -1 ) Apprent Digestiity Coefficient (%) () Soy1 Soy2 Soy3 Soy4 Control Comm Tretments c (d) Soy1 Soy2 Soy3 Soy4 Control Tretments d c Figure 1-d. Different prmeters evluted on juveniles of rinow trout fed diets with high levels of soyen mel. () Oxygen consumption. () Phosphorus excretion. (c) Nitrogen excretion. (d) Apprent digestiility coefficient. Vlues re mens of the three replicte groups ± stndrd devition. Mens with different letters in the sme column differ significntly (p 0.05). Comm refers to the commercil diet. The N excretion of the fish fed with diets Soy2, Soy3 nd Soy4 showed significntly lower vlues thn those oserved in the commercil nd the control diet nd in the diet with sustitution of 50% of soyen mel (Fig. 1c). Higher vlues of ADC were oserved in the diets with soyen mel sustitution, prticulrly with totl fishmel replcement. The control diet with 100% fishmel showed the lowest vlues of ADC (Fig. 1d). DISCUSSION One of the min chllenges of quculture industry for the yers to come is to find lterntive ingredients, which re cheper nd environmentl friendly (Gtlin et l., 2007). Soyen mel hs een pointed out s n lterntive source of protein for qufeeds (Hrdy, 1996), ecuse of its protein content nd resonle welllnced mino cid profile. However, its use t high levels of supplementtion (greter thn 50%) hs een discourged due to the presence of some nti-nutritionl compounds, prticulrly the content of phytic cid. This compound hs een reported to e indigestile nd to cuse negtive effects on growth, feed conversion nd mcronutrient digestiility (Lining et l., 2010). Previous reports hve suggested tht the use of the enzyme phytse (Cheng et l., 2004; Wng et l., 2009) llows to improve the digestiility of diets nd thus, growth. The dt otined in the present reserch show tht soyen mel might e used up to Hidroiológic Soyen mel in diets of rinow trout % without ffecting significntly the growth performnce of juvenile rinow trout when 0.8 g kg -1 of phytse is dded to the diet. The fish fed this diet (Soy2), showed growth performnce similr to tht reported y Wng et l. (2009) in rinow trout of n initil weight of 1.7 g nd fed diet with 60% of soyen mel s source of protein. As mentioned efore, growth of fish is relted to the digestiility of the diet nd prticulrly to tht of the protein frction. The ADC of protein of the diets with soyen mel ws higher thn the reported y Wng et l. (2009) nd Dlsgrd et l. (2009), ut coincides with those reported y Cheng et l. (2004). It seems tht phytse incorportion into diet is relted to the digestiility of protein portion. When dded fter diet preprtion (Dlsgrd et l., 2009) or is spryed (Wng et l., 2009) on the diet, ADC tends to e lower thn when the phytse is dded efore preprtion, s in this study. Interestingly, fish fed with diets Soy3 nd Soy4 (100% of sustitution of fishmel) showed the highest vlues of ADC of protein. In ddition, those diets showed less excretion of NH 3 -N nd oxygen consumption. According to Bureu et l. (2002), N excretion nd oxygen consumption my e used to estimte the oxidtion of nutrients from the diet nd, usully, low vlues indicte etter use of the protein source (less excretion) nd less energy spent in its oxidtion (less consumption). However, fish fed those diets showed significntly lower growth thn tht oserved with Soy1, Soy2, control nd commercil diets. Soyen is deficient in lysine, methionine nd threonine (Gtlin et l., 2007), three essentil mino cids. This proly cused the poor growth performnce in 100% soyen mel sed diets, even if these hd the highest digestiility of ll diets. The fishmel inclusion in the other diets could llow n increse in the levels of such mino cids nd etter growth performnce. Phosphorus plys n importnt role in fish, ecus
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