Effect of Dietary Lipid Levels on Fatty Acids Composition of Cultured Sparusaurata

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1 Journl of Life Sienes 10 (2016) doi: / / D DAVID PUBLISHING Effet of Dietry Lipid Levels on Ftty Aids Composition of Cultured Sprusurt Zyene Nesrine 1,2, Gm Hous Wf 1,2, Guerej Hmdi 2 nd Ahour Lotfi 1 1. Integrtive Biology nd Vloristion (LR 14 ES 06), Institute of Biotehnology, Monstir 5000, Tunisi 2. Ntionl Institute of Mrine Sienes nd Tehnologies, Monstir 5000, Tunisi Astrt: The im of this study ws to determine the progressive evolution of fish musle ftty ids omposition feeding y different lipid levels of frmed Mediterrnen gilthed se rem (Sprusurt). During twenty one weeks, Sprusurt were hnd-feed two diets with protein nd lipid rtios 47/20 (Diet A) nd 45/22 (Diet B) respetively. At the end of the feeding period, lipid ontents of musle, s well s ftty ids profiles of different portions of the musles were studied. Results indited the effets of ftty ids profiles of diets throughout the liver nd white musle fillet (dorsl, middle nd ventrl). These were more notiele when Sprusurt ws fed on the diet Atht deresed the ontent of monounsturted ftty ids (MUFA), nd polyunsturted ftty ids (PUFA) in ll the musles during the period of feeding while the sturted ftty ids (SFA) ontent tended to inrese nd it ws lower thn ones in the musles of Sprusurt fed on diet B. The highest mount of PUFA nd dooshexenoi id, (22: 6n-3 DHA) were found mong the different portions of ventrl musles. In ddition, the progressive evolution of different prts of musle s ftty id profiles showed signifint differenes in ontents of SFA, PUFA nd espeilly the n-3 PUFA. However, the ontent of MUFA seems to e like in different musles nd tended to derese in dorsl nd ventrl musle from D 0 to D f nd inversely when the fish were fed on diet A. Key words: Ftty ids, Liver, Musles, ultured fish, Sprusurt. 1. Introdution In reent yers, mny studies reported the high enefits of onsumption of mrine oils in humn helth ording to Simopoulos 2003 [1]; Shmidt et l., 2005 [2] nd Kremer 2000 [3]. Coiniding with this nd the overexploittion of fishing grounds in World nk [4], quulture tivities hve stirred growing interest. In the Mediterrnen re, notle suess hs een hieved in the prodution of diverse speies, suh s se rem (Sprusurt) nd se ss (Dientrrhuslrx). Therefore, rering these speies hs een redireted with the improvement of the finl eting qulity of the produt. The se rem (Sprusurt) is onsidered to e very importnt quulture fish speies. On the other hnd, suh s mrine fish, Sprusurt is onsidered s soure of highly unsturted ftty ids fmily Corresponding uthor: Zyene Nesrine, Ph.D., reserh field: fish nutrition. (HUFA) [5]. However, the types nd mounts of ftty ids in fish tissues vry with the geogrphi lotion, size, ge, quulture feeds, reprodutive sttus nd sesons [6-9]. The short or long period of feeding (period of rering to hve mrketle ommeril size) my, lso, influene the ftty ids profile of the fillet qulity s is shown in mny rered fish speies [10, 11]. The ojetive of this study ws to evlute the effets of two lipids levels of extruded diets on ftty id omposition of different edile musles of rered Sprusurt t different dtes during the period of reeding. 2. Mterils nd Methods 2.1 Diets nd Feeding The study ws sed on two dietry experiments. The experiene ws performed t Ntionl Institute of Mrine Sienes nd Tehnologies of Monstir, Tunisi;

2 Effet of Dietry Lipid Levels on Ftty Aids Composition of Cultured Sprusurt 313 from Ferury 2009 to July 2009 (totl 21 weeks). Two ommeril diets ontined two different lipids levels (20% nd 22%) were used for feeding the juvenile Sprusurt. At the strt of the feeding tril, two groups of 180 fish of ± 3.48 g men ody weight were rndomly weighed nd stoked into two round tnks. The fish were fed to stition y hnd, twie dy. Eh diet ws rndomly lloted to duplite groups. 2.2 Smpling Proedure Smplings were performed regulrly fter 21 dys, so fish from eh tnk were weighed t first dy 0 (D 0 ), then fter 11 weeks (D i ) nd fter 21 weeks (D f ). From eh diet, five fish were smpled nd pproximtely 0.5 g of musle from different edile prts of fish (dorsl musle: DM, middle musle: MM nd ventrl musle: VM) nd the liver were disseted out nd stored t -80 C for susequent lipids nd ftty ids nlyses. 2.3 Lipids nd Ftty Aids Determintion Lipids for ftty id nlysis were extrted from diets nd different musle smples with hloroform nd methnol ording to the proedure of Folh et l., (1957) [12], then methylted nd trnsesterified with oron trifluoride in methnol [13]. The internl stndrd (19: 0) ws dded t 1% of the totl lipid weight for quntittive mesurement of individul ftty ids present in the diets nd musles. Ftty id methyl esters were resolved nd nlyzed y Hewlett-Pkrd HP 5890 pillry gs hromtogrph linked to n HP Chemsttion integrtor. The pillry olumn ws of HP-Innowx sili (30 m long, 0.25 mm internl dimeter with 0.25 µm thikness of film of the sttionry phse). The quntittive nlysis of the ftty ids ws mde y ompring the retention times of the ftty ids in the hromtogrms orresponding to the retion produts, with those of stndrd mixture, from Sigm Chemil Co. 2.4 Sttistil Anlysis Dt otined were ll sujeted to one-wy nlysis of vrine (ANOVA) to test the effets of different dietry lipid soure nd the reeding period on ftty id omposition of different musles. Differenes etween mens were determined y Dunn s Multiple Rnge test nd were onsidered to e signifint when P < All nlysis ws performed using the PASW sttistis 18 for Windows. 3. Results nd Anlysis 3.1 Ftty Aid Composition of the Diets The nlyti nd ftty id omposition of the ommerildiets were shown in Tles 1 nd 2. Aording to the results of nlyses, there were signifint differenes mong the levels of different ftty ids. Tle 1 Ingredients nd nlytil omposition of the two ommeril diets. Diets Diet A Diet B Fish mel Fish oil Soyen mel Fish mel Fish oil Soyen mel Ingredients Orgni pes Vitmin premix Minerl premix Rpeseed mel Orgni pes Vitmin premix Minerl premix Anlytil omposition Crude protein (%) Crude lipid (%) Crude fier (%) Ash (%)

3 314 Effet of Dietry Lipid Levels on Ftty Aids Composition of Cultured Sprusurt Tle 2 Ftty id profile (% of totl ftty ids) of the ommeril diets (wet weight). Diets Diet A Diet B C14:0 1.76± ±0.27 C15:0 0.24± ±0.06 C16: ± ±0.99 C17:0 0.34± ±0.05 C18:0 2.22± ±0.03 C20:0 0.38± ±0.02 C22:0 1.90± ±0.16 C24:0 1.21± ±0.05 SFA 51.21± ±0.47 C14:1 0.49± ±0.09 C15:1 0.36± ±0.04 C16:1 1.92± ±0.04 C17:1 0.30± ±0.03 C18:1n ± ±0.02 C18:1n ± ±0.06 C20:1n ± ±0.04 C22:1 1.55± ±0.11 C24:1 0.16± ±0.02 MUFA 19.85± ±0.23 C18:2n ± ±0.41 C20:2n ± ±0.06 C20:3n ± ±0.02 C20:4n ± ±0.11 C22:4n ± ±0.18 n-6 PUFA 17.79± ±0.66 C18:3n ± ±0.34 C20:5n-3 (EPA) 2.93± ±0.06 C22:5n ± ±0.02 C22:6n-3 (DHA) 5.06± ±0.10 n-3 PUFA 10.98± ±0.16 PUFA 28.77± ±0.50 Vlues re the verges of three replites ±sem. Numers in rows hving different letters indite tht tretment re signifintly different. The two diets hd similr verge of SFA ( 50%) nd PUFA ( 28%) with n importnt level of n-6 PUFA ( 15 %) respetively (Tle 2). Compring etween two diets, the diet B ontined higher proportions of SFA nd n-3 PUFA. Wheres the diet A hd higher proportion of shorter 18C hin-length unsturted ftty ids nd n-6 PUFA. In the two diets, plmitind steri ids were the primry sturted ftty id (SFA), ontriuting pproximtely 90% to the totl SFA ontent of the lipids. Olei id ws identified s the primry monoenoi ftty id nd ws signifintly (P < 0.05) higher in diet A thn in B. Among n-6 series of the ftty ids, linolei id (18: 2n-6) hd higher level. This ftty id is present inplntoils used in the feed ingredient of ultured fish [14-16] Among the n-3 series, oth diets were good soures of EPA nd DHA. The perentges of EPA nd DHA were (2.93% in diet Avs 3.06% in diet B) nd (5.06% in diet A vs 6.30% in diet B) respetively. 3.2 Ftty Aids Composition of the Liver The ftty id profiles of liver of the two groups of

4 Effet of Dietry Lipid Levels on Ftty Aids Composition of Cultured Sprusurt 315 ultured Sprusurt re presented in figures 1 ( nd ). The perentge of totl sturted ids ws higher in the group of fish fed on diet B thn on diet A, wheres its totl monoenoi nd polyenoi ontent ws lower. The mjor polyunsturted ftty ids identified in oth group of fish were 20: 5n-3 (eiospentenoi id, EPA) nd 22: 6n-3 (dooshexenoi id, DHA). Liver of ultured se rem ontined signifintly (P < 0.05) higher proportions of n-3 PUFA nd speilly higher proportion of DHA thn n-6 PUFA. At the sme time, the figure 1 showed the evolution of ftty ids vlues t the D i nd D f. The vlues of different groups of ftty ids were, on verge, similr regrdless the dte of smpling Do Di Df 50 % of ftty ids d e e d SFA MUFA PUFA PUFA n 6 PUFA n 3 C22:6 ω 3 (DHA) e d e d e d C20:5 ω 3 (EPA) () Sprusurt fed on diet A with 20% of lipid. 70 % of ftty ids de d SFA MUFA PUFA PUFA n 6 PUFA n 3 C22:6 ω 3 (DHA) d Do Di Df C20:5 ω 3 (EPA) () Sprusurt fed on diet B with 22% of lipid. Fig. 1 Ftty ids omposition (% of totl ftty ids) nd distriution in the liver of eh group of Sprusurt fed on diet A nd diet B. Vlues re the verge of three replites ±sem. Numers in histogrms hving different letters indite tht tretment re signifintly different P < 0.05.

5 316 Effet of Dietry Lipid Levels on Ftty Aids Composition of Cultured Sprusurt But sttistil nlyses reveled tht during the first experimentl dys (D i ), differenes were not very importnt in SFA profiles inliver ompred etween the initil profile (D 0 ) nd the finl one t D f. But, MUFA nd PUFA tended to derese (from 33.91% to 28.91% for the MUFA nd from 35.16% to 31.58% for the PUFA) in the liver of the fish fed on diet B nd inversely in the fish fed on diet A Ftty Aids Composition of Different Edile Musles. The ftty id ompositions of different ody prts (the dorsl, middle nd ventrl musles) of Sprusurt re presented in figures 2 (, nd ) respetively. Compring ftty ids lss s level in eh group of gilthed serem, vlues showed tht ll musles hd signifintly higher levels of SFA nd PUFA thn the MUFA levels. Moreover, mong the PUFA group, the n-6 PUFA levels were signifintly lower thn n-3 PUFA whih een expressed essentilly y the DHA id. Musleftty id omposition of totl lipids showed the effet of the diets from the intermedite smpling (D i ) to the end of feeding diets (D f ) when fish rehed the ommeril size. Sturted ftty ids (SFA) inresed from the eginning smpling until the end of feeding experiene (D f ). However, MUFA deresed in themusle of the fish fed on diet A nd B in the intermedite smpling ut did not further inrese in the finl smpling (Figs. 2, nd ). Moreover, PUFA deresed long the first period in the diets A nd B proportionlly to their level in the diets ut up to the level of 40% of totl ftty ids, totl PUFA derese t the end of experiment. In omprison etween fish fed on diet A nd B, n-3 PUFA musleontents in the first smpling redued y 3.5% in the verge in ll musles. Then, the levels of these ftty ids were further redued until the end of the experimentl period rehing 13% nd 20% redutions in fish fed 20% nd 22%. Redution of ftty ids long the diet B ws more pronouned fordha thn EPA in the intermedite nd in the finl smpling (Fig. 2). d e d Σ SFA Σ MUFA Σ PUFA Σ PUFA n-6 Σ PUFA n-3 C 22:6 n-3 DHA C 20:5 n-3 EPA () Dorsl musle (DM) of Sprusurt fed on diet A (20% of lipid) nd diet B (22% of lipid).

6 Effet of Dietry Lipid Levels on Ftty Aids Composition of Cultured Sprusurt % of ftty ids d d d MM 20% MM 22% D0 DI DF D0 DI DF D0 DI DF D0 DI DF D0 DI DF D0 DI DF D0 DI DF d Σ SFA Σ MUFA Σ PUFA Σ PUFA n-6 Σ PUFA n-3 C 22:6 n-3 DHA C 20:5 n-3 EPA () Middle musle (MM) of Sprusurt fed on diet A (20% of lipid) nd diet B (22% of lipid). 70 % of ftty ids d d VM 20% VM 22% D0 DI DF D0 DI DF D0 DI DF D0 DI DF D0 DI DF D0 DI DF D0 DI DF d d d Σ SFA Σ MUFA Σ PUFA Σ PUFA n-6 Σ PUFA n-3 C 22:6 n-3 DHA C 20:5 n-3 EPA () Ventrl musle (VM) of Sprusurt fed on diet A (20% of lipid) nd diet B (22% of lipid). Fig. 2 Ftty ids omposition (% of totl ftty ids) nd distriution in the Dorsl musle (MD)--, Middle musle (MM)-- nd Ventrl musle (MV)-- of eh group of Sprusurt fed on diet A (with 20% of lipids) nd diet B (with 22% of lipid) from initil to finl period of experiment. Vlues re the verge of three replites ±sem. Different letters in histogrms indite tht tretment re signifintly different P < 0.05.

7 318 Effet of Dietry Lipid Levels on Ftty Aids Composition of Cultured Sprusurt 4. Disussion In the present study, fish were fed on two diets (20% nd 22% lipid levels) during 21 weeks to mrketle size fish. As suh, the deposition of ftty ids in liver nd different nlysed musles of Sprusurt ws influened y the ftty id omposition of the experimentl diets. In generl, ftty ids found in high onentrtions in the diet were lso refleted to ertin extent in the ftty id omposition of musle tissue. These results re in greement withother studies reporting tht the ftty id pttern of fish tissue reflets tht of dietry lipids [17, 18]. In this study, there re mrked differenes in ftty id deposition in different musles of ultured Sprusurt. Generlly, thedorsl nd middle prts (DM nd MM) hd higher onentrtion of SFA nd PUFA, espeilly in fish fed on 22% of lipid level. In omprison etween the two groups, the musle of the fish fed on diet A, ontined signifintly higher level of PUFA, n-6 PUFA nd MUFA. Similr to these, high proportions of DHA were lso oserved in the dorso-ventrl musle. However, DHA ontent ws doule tht of EPA nd ws not inresed y the feeding with the two diets for 21 weeks. Moreover, feeding with diet A (20% of lipid level) for 11 weeks effetively inresed DHA ontents in musle of fish, improving the nutritionl vlue of serem fillets for humn [19]. But EPA s fillet ontents were not fully reovered even fter 21 weeks, this ftty id eing lso very importnt for humn helth, s potent hypotriglyeridemi ftor [20] nd more effetive thn DHA in inhiiting pltelet ggregtion [21]. So, s it shown, these results re similr to the results found on gilthed serem fed on vegetle oils for long term period [22], signifintly higher level of SFA, MUFA nd PUFA were stored in the flesh ompred to the eginning of diet experimentdue to the introdution of ertin ftty ids of vegetl origin suh s α-linoleni id in formultion of fish diets. In omprison with the evolution of DHA musle ontents, this higher redution of DHA nd lower inorportion of EPA fter feeding with diets A nd B my e relted with severl ftors. On one hnd, it ould e due to preferentil oxidtion of EPA over DHA in greement with the higher redution of EPA in the neutrl lipids, sine white musle seem to ply key role in the overll ftty id oxidtion pity in fish, mitohondril et-oxidtion dominting over peroxisoml oxidtion in this tissue [23], nd EPA is minly oxidized y mitohondri, wheres DHA seems to e oxidized y the peroxisomes nd to lower extent thn EPA [24]. On the other hnd, there is high ffinity of phosphtidylholine (PC) nd, espeilly, phosphtidylethnolmine (PE) synthetses for DHA, prtiulrly in the 2n position, nd in gilthed se rem lrve elevtion of dietry DHA inhiits EPA inorportion into PE, wheres elevtion of dietry EPA levels enhnes DHA inorportion into PC nd PE [25]. In greement with those uthors, in the present experiment the elevtion of DHA enhned the inorportion of DHA rther thn tht of EPA. From strt-feeding to mrketle size of fish, the higher proportion of SFA nd PUFA ppered to e the retention nd storge ftty ids, suh s the se DHA. However, MUFA nd n-6 PUFA were eing the tively synthesized euse they were limited in musle tissue ut undnt in oil diets. Thus, there mining ftty ids found in oth speies (out 70%) were sturted nd polyunsturted ftty ids (SFA+ PUFA). These vlues re higher thn the reported vlues (15.7%) in silver rp (Hypopththlmiehthysmolitrix) [26]. In generl, fish re reltively low in sturted ftty id (< 30%), exept for ertin speies [27]. Similr results for wild znder [28] nd other freshwter fish hve lso een reported in the literture [26].

8 Effet of Dietry Lipid Levels on Ftty Aids Composition of Cultured Sprusurt 319 The sturted ftty id ontent didn t remin onstnt nd inresed mongst the different fillet portions nd/or the different fish sizes. This is in greement with studies of Turhini et l. (2003) [29] nd Frnis et l. (2006) [30] in tht SFA re not used effiiently y gilthed se rem (Sprusurt) s n energy soure nd re therefore umulted t n optiml level ompred to other ftty id lsses. On the ontrry to SFA, there ws derese in the PUFA ontent nd n inrese in tht of MUFA in the musle for the first dys. Overll, in highly ft-rih fillet portions n-6 ftty ids were less undnt, wheres n-3 were predominnt, suggesting tht n-3 re preferentilly deposited s stored lipid, while n-6 ftty id re importnt prt of the funtionl lipid. The eiospentenoi id ontent remined firly uniform in the different fillet portions, suggesting tht its perentge ontent is similr in oth stored nd funtionl lipids. In fish, generlly, umultion of ertin ftty ids in musle tissue is dependent on their dietry onentrtion. However, in this study, the n-3 PUFA vlues were higher in musle thn in the diet. The sme trend hs een oserved in slmonids relesed y Turhini et l., 2003 [29], suggesting tht Sprusurt tend to umulte nd store n-3 while using n-6 s n energy soure. 5. Conlusions In summrising, it my e onluded tht the period of reeding of Sprusurt fed on medium differenes of lipid s diet levels (20% in the diet A) nd (22% in the diet B), did not produe diverge in terms of ontents of EPA nd DHA ids, the most vlule to onsumer, or in the vlues of suh inditors of the nutritionl qulity of lipids s n-3/n-6 nd EPA/DHA rtios. Nevertheless, there ws n inresed ft deposition s the fish size inresed, suggesting tht Sprusurt utilises lipid t fster rte during erly growth stges, nd then strts umulting ft s growth dereses. This is in ordne with tht desried y Bell et l. (2003) [31] nd Plmeri et l. (2007) [32] for Atlnti slmon nd Murry od. Despite these hnges in musle ftty id omposition, fish fillets remined eing highly nutritious for humn helth. Referenes [1] Simopoulos, A. P Importne of the rtio of omeg-6/omeg-3 essentil ftty ids. Evolutionry spets. In AP Simopoulos & LG. Clelnd (Eds.), Omeg-6/Omeg-3 essentil ftty id rtio: The sientifi evidene (pp. 1-22). Bsel, CH: Krger. [2] Shmidt, E. B., Arnesen, H., Cterin, R., Rsmussen, L. H., nd Kristensen, S. D Mrine n-3 polyunsturted ftty ids nd oronry hert disese. Prt I. Bkground, epidemiology, niml dt, effets on risk ftors nd sfety. Throm Res. 115 (3): doi: /j.thromres [3] Kremer, J. M N-3 ftty id supplements in rheumtoid rthritis. Am. J. Clin. Nutr. 71: 349S-51S. [4] World Bnk World Bnk Group Agriulture Ation Pln Wshington, DC: World Bnk. FISH TO 2030: Prospetus for fisheries nd quulture. [5] Mourente, G., nd Toher, D. R The in vivo inorportion nd metolism of [1-14C] linolente (18:3n-3) in liver, rin nd eyes of juveniles of rinow trout Onorhynhusmykiss L nd gilthed se rem Sprusurt L. Fish Physiology nd Biohemistry 18: [6] Bndrr, N. M., Nunes, M. L., Andrde, A. M., Prtes, J. A. M., Pereir, S., Monteiro, M., Rem, P., nd Vlente, L. M. P Effet of dietry onjugted linolei id on musle, liver nd viserl lipid deposition in rinow trout juveniles (Onorhynhusmykiss). Aquulture 254: [7] Çelik, M., Diler, A., nd Küçükgülmez, A A omprison of the proximte ompositions nd ftty id profiles of znder (Snder luioper) from two different regions nd limti onditions. Food Chem. 92: [8] Rued-Jsso, R., Coneio, L., Dis, J., De Coen, W., Gomes, E., nd Rees, J Effet of dietry non-protein energy levels on ondition nd oxidtive sttus of Seneglese sole (Soleseneglensis) juveniles. Aquulture 231: [9] Grigorkis, K., Alexis, M. N., Tylor, A., nd Hole, M Comprison of wild nd ultured gilthed se rem, (Sprusurt): omposition, pperne nd

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