DIET EVALUATION AND ESSENTIAL FATTY ACIDS INCORPORATION IN KURUMA PRAWN, Penaeus japonicus

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1 J. Agrolnd 13 (3) : , September 26 ISSN : X DIET EVALUATION AND ESSENTIAL FATTY ACIDS INCORPORATION IN KURUMA PRAWN, Peneus jponicus By: Ek Rosyid 1) ABSTRAK Penelitin ini bertujun untuk mengethui pengruh komposisi sm lemk (ftty cid) dlm mknn terhdp pertumbuhn dn kelngsungn hidup post lrv (PL) udng kurum, Peneus jponicus. Dismping itu, komposisi dn meknisme trnsfer sm lemk diuji sesui urutn rnti mknn: lge, Artemi dn udng. Hsil penelitin menunjukkn bhw bobot bdn (Body Weight=BW) tertinggi dicpi oleh udng yng mengkonsumsi Artemi pemkn lge, Isochrysis glbn (T.Iso) (P<,5). Sedngkn untuk pnjng totl (Totl Lenght=TL) tertinggi ditunjukkn oleh udng-udng yng mengkonsumsi Artemi pemkn lge, T.Iso dn yng mengkonsumsi Artemi pemkn lge, Tetrselmis suecic. Mknn lmi Artemi lebih bik untuk pertumbuhn lrv udng dibnding mknn butn. Artemi dpt mentrnsfer zt-zt gizi khususny sm lemk yng terdpt dlm lge ke udng. Dlm kedn lpr, ktbolisme sm lemk terjdi dlm tubuh udng, dn sm lemk gnd tk jenuh omeg 3 lebih efektif untuk mendukung pertumbuhn udng dibnding omeg 6. Keseimbngn komposisi ntr berbgi sm lemk gnd tk jenuh kemungkinn sngt penting untuk pertumbuhn udng. Kt kunci : Udng kurum, lge, Artemi, sm lemk, pertumbuhn. ABSTRACT A 4-weeks experiment ws undertken to study effects of dietry ftty cid composition of post lrvl growth nd survivl of Peneus jponicus. Ftty cid composition nd trnsfer mechnism were lso exmined long the following food chin: lge Artemi prwn. Results indicted tht the highest body weight gin ws observed for the P. jponicus fed on Artemi consuming lge Tetrselmis iso, while the highest totl length ws indicted by the prwn fed on Artemi consuming both T. iso nd T. suecic. It ws lso shown tht the live feed ws superior for the post lrvl growth of P. jponicus compred to commercil feed, nd tht the Artemi ws good in trnsferring importnt nutrients from lge to the prwn. The body ftty cid ws shown to be ctbolized when the prwn ws in strved condition. Polyunsturted ftty cids (PUFAs) of the n3 series ppered to be more effective in supporting the post lrvl growth of P. jponicus which my indicte tht the n3 series FA is more importnt thn the n6 series. Key words : Peneus jponicus, lge, Artemi, ftty cids, growth I. INTRODUCTION The potentil nd economic importnce of kurum prwn, Peneus jponicus hs ttrcted interest of this species in quculture industry. This peneid species hs unique requirements for cholesterols nd phospholipids with regrd to the growth nd development, nd these re indictive of the importnce of dietry lipid to peneid growth nd development (D Abrmo, 1997). 1 ) Stf Pengjr pd Progrm Studi Budidy Perirn Fkults Pertnin Universits Tdulko, Plu. The nutritionl vlue of dietry lipids is prticulrly determined by their ftty cid composition in reltion to metbolic processes such s ctbolism nd storge of energy. It is well documented tht most of mrine nimls require specific dietry ftty cids (FAs), such s eicospentenoic cid (EPA, 2:5n3) nd docoshexenoic cid (DHA, 22:6n3) for good growth nd survivl during lrvl nd juvenile stges. Unfortunetly, the peneid prwns hve only limited cpbility for de novo synthesis of certin essentil ftty cids 36

2 (EFAs) nd so must obtin them from the diet to chieve good growth (Knzw, 1985; Mericn nd Shim, 1997). D Souz nd Lonergn (1999) concluded tht the survivl nd development of prwn lrve ws influenced by the distinctly different FAs profile in ech species of lge used. Brown et l. (1989) indicted tht FAs of most lge comprise up to 4 % of the totl lipid (weight bsis) nd these FAs re suitble to meet prwn lrve lipid s requirements. Following the erlier studies on P. monodon (Rosyid, 22), in the present study, the growth nd the survivl rte of P. jponicus were lso exmined bsed on vrious diets i.e Tetrselmis suecic, Isochrysis glbn (the Thitin strin, referred to s T.Iso), Dunliell tertiolect, Brine shrimp (Artemi) nd commercil diet. On the other hnd, the cpbility of Artemi to trnsfer some vluble FAs from different species of microlge into P. jponicus post lrve (PL) were ssessed. A commercil prwn diet ws used s control nd its FA composition ws lso ssessed for comprison with the Artemi diets. II. MATERIALS AND METHODS 2.1 Culture of lge nd Artemi The three species of lge used : Dunliell tertiolect, Isochrysis glbn (Thitin clone) nd Tetrselmis suecic were cultured under semi continuous btch culturing system (s described in O'Melley nd Dintith,1993) in the 'Key Center', School of Aquculture, Univ. of Tsmni. Artemi used in these trils were AS (Artemi Systems, Belgium), INVE Aquculture. The cysts were incubted in filtered sewter using conicl continers t o C under continuous ertion nd light using the methods of Dintith (1996). Freshly htched Artemi nuplii (36 hours old) were rised on the 3 different species of lge for 6 or 8 dys. Nuplii were suspended t density of 1/ml in erted tnks (1 L volume) nd fed lge twice dily t density of t lest 1 x 1 5 cells/ml. After 6 or 8 dys, the Artemi were hrvested onto 15 m screen, thoroughly rinsed with sterile filtered sewter, nd counted by microscopic exmintion using Sedgwick Rfter slide nd poured into ice cube trys before being frozen. 2.2 Experimentl nimls nd design P. jponicus PL used in this experiment were obtined from the Rocky Point htchery, Woongoolb, Queenslnd. The men initil weights (W) of individul lrve ws 1.2 mg nd the men totl length (TL) ws 3.1 mm. The experiments were conducted in recirculting system for 4 weeks. Fifteen 52 L drk tnks were used, ech with flow rte of 3 ml/min. Aertion ws provided by one irstone in ech tnk, nd wter qulity ws mesured nd checked every week. Wter temperture ws mintined t 26 to 27 C by heter chiller unit (Acktron 7.9 kw) nd slinity of 33±1 / ws mesured nd djusted dily when sltwter ws dded in the sump. Five experimentl tretments run in triplicte : three Artemi diets rised on different lg species, one control tretment where prwns were fed commercil diet (Fry No.T 1 with protein nd ft content of 4 % nd 4.5 %, respectively), nd one tretment where the nimls were unfed. The prwns were stocked t 5 PL in ech tnk (1 PL/L), nd were fed three times dily, t 8, 12 nd 17 h. Artemi were given in the mount of 1 Artemi/PL/feed, while the commercil diet ws provided t.1 g/tnk/feed. Uneten diet from ech tnk ws gently siphoned through smll irline fitted with Psteur pipette t the siphon end tht contcted the wter to ensure tht no prwns were sucked out. 37

3 2.3 Smpling for ftty cid determintions The experimentl procedures showing the mechnisms of trnsferring diets to the prwns is presented in the following digrm. As lge fed to Artemi everydy, the smples were tken for severl times to estimte the verge. Smples of lge were collected by filtering pproximtely 25 ml of ech species onto Whtmn glssfibre filters (47 mm) which hd been precombusted for 4 45 C. Smples nd filters were then folded in hlf nd plced into luminium foil pockets. Artemi were smpled t ech ge fed to prwns (6 nd 8 dys old) nd the necessry mount ws plced into luminium pockets. Seven to ten prwns from ech tretment were rndomly smpled t the beginning nd t the end of ech experiment nd then plced into foil pockets. All smples were lbelled nd seled in plstic bgs filled with nitrogen gs, nd stored in - 8 C freezer until further nlysis. Ftty cid methyl esters (FAMEs) were prepred from the smples by sponifiction, then followed by methyltion s described in Whyte (1988). At the end, the extrction results stored in freezer until further nlysis. 2.4 Smpling for growth determintions Thirty PLs were rndomly selected from the stock popultion for initil weight (W) nd totl length (TL) determintions, while ten to twenty prwns were smpled from ech tretment for the sme mesurements t the end of experiment. Totl survivl ws lso clculted t the end. 2.5 Ftty cid nlysis nd Sttisticl nlysis Anlysis of ftty cids (FA) ws undertken t CSIRO-Mrine Lbortories, Hobrt, Tsmni. Gs chromtogrph (GC) nlysis of FAMEs were performed nd detils of this GC re provided by Nichols et l. (1989). The effect of diet on growth nd survivl were determined by one wy nlysis of vrince (ANOVA) using SPSS version 9.. Post hoc comprisons were undertken with Tukey HSD, nd the results were considered sttisticlly significnt t the P.5 level. Ftty cids of phytoplnkton, Artemi nd prwns were nlysed sttisticlly using ANOVA nd followed with Kruskl Wllce ANOVA on rnks. T. suecic T. iso D. tertiolect Artemi 6 dys old Artemi 8 dys Artemi 6 dys Artemi 8 dys Artemi 6 dys Artemi 8 dys Commercil diet No diet Prwns Prwns Prwns Prwns Prwns Figure 1. The Digrm of Experimentl Procedures Showing The Mechnisms of Trnsferring Diets to The Prwns, P. jponicus. 38 1

4 III. RESULTS AND DISCUSSION 3.1 Growth nd survivl There ws no significnt difference in the survivl of mongst Kurum prwn fed different diets (P>.5) (Fig. 2). Weight gin ws gretest when the prwns were fed on Artemi rised on T. iso (P<.5) (Fig. 2b). On the other hnd, there ws no significnt difference mongst prwns fed on the other 3 diets (P>.5). When the prwns were left unfed, however, they grew significntly slower compred to ny other tretments (P<.5). Survivl (%) weight gin (mg) Totl length (mm) A B C Ar-Ts Ar-Ti Ar-Dt Com unfed bc Ar-Ts Ar-Ti Ar-Dt Com Unfed b Ar-Ts Ar-Ti Ar-Dt Com unfed Diet c bc Figure 6. Survivl (A), weight gin (B) nd totl length (C) of P. jponicus fed on vrious diets. Ar-Ts= Artemi rised on T. suecic, Totl Ar-Ti= Length Artemi (C) rised of on P.jponicus T.Iso, Ar-Dt= fed Artemi rised on D. tertiolect nd Com= commercil diet. Verticl on brs vrious represent stndrd diets: devition. Ar-Ts Vlues = Artemi tht shre the sme superscript re not significnt different. Figure 2. Survivl (A), Weight gin (B) nd rised on T.suecic, Ar-Ti = Artemi rised on T.Iso, Ar-Dt = Artemi rised on D.tertiolect nd com=commercil diet. Verticl brs represent stndrd devition. Vlues tht shre the sme superscript re not significnt different. c bc d d 1 The prwns fed on Artemi rised on T. suecic nd T. Iso obtined the longest totl 39 length (Fig. 2c), which incresed lmost threefold over the experimentl period. The totl length between prwns fed on Artemi rised on D. tertiolect nd commercil diet were similr (P>.5), while unfed prwns incresed only 1 mm of the totl length which ws significntly less compred to ll fed prwn tretments (P<.5). 3.2 Ftty Acid Composition of Alge, Artemi, commercil diet nd prwn, P. jponicus FA composition of lge, Artemi, commercil diet nd prwn, P.jponocus used in this tril vried considerbly s shown in Fig.3 dn Fig.4. Proportion ( % of totl ftty cid) : 16: 18: 16:1n7 18:1n9 18:1n7 18:2n6 18:3n3 2:4n6 2:5n3 Mjor ftty cids Ar-Ts Ar-Ti Ar-Dt Com 22:6n3 Figure 7. The proportion of mjor ftty cids in P. jponicus diets. Ar-Ts= Artemi rised on T. suecic, Ar-Ti= Artemi rised on T. Iso, Ar-Dt= Artemi rised on D. tertiolect nd Com= commercil diet. Verticl brs represent stndrd devition. Figure 3. The proportion of mjor Ftty Acids in P.jponicus diets. Ar-Ts=Artemi rised on T.suecic, Ar-Ti=Artemi rised on T.Iso, Ar-Dt=Artemi rised on D.tertiolect nd com= commercil diet.verticl brs 3 represent stndrd devition. Propotion (% of totl ftty cid) : 16: 18: 16:1n7 18:1n9 18:1n7 18:2n6 18:3n3 Mjor ftty cids 2:4n6 2:5n3 KI KAr-Ts KAr-Ti KAr-Dt KCom Unfed 22:6n3 Figure 8. The proportion of mjor ftty cids in the kurum prwns, P. jponicus. KI= prwns initil, KAr-Ts= prwns fed on Artemi rised on T. suecic, KAr-Ti= prwns fed on Artemi rised on T. Iso, KAr-Dt= prwns fed on Artemi rised on D. tertiolect nd KCom= prwns fed on Artemi rised on commercil diet. Verticl brs represent stndrd devition. Figure 4. The proportion of mjor ftty cids in the kurum prwns, P.jponicus. KI=prwns initil, KAr-Ts=prwns fed on Artemi rised on T.Suecic, KAr-Ti=prwns fed on Artemi rised on T.Iso,KAr-Dt= prwns fed on Artemi rised on D.tertiolect nd KCom=prwns fed on Artemi rised on commercil diet. stndrd devition. Verticl brs represent

5 The present study showed tht the lck of linolenic cid, 18:3n3 my effect the growth of P. jponicus PLs. Even though the linoleic cid (18:2n6); EPA (2:5n3) nd DHA (22:6n3) were greter in the tissue of the prwns fed on the commercil diet compred to the other fed prwns, the ner bsence of linolenic cid my hve resulted in the slowest growth of these prwns. The essentility of these four FAs in lrv nd juvenile P. jponicus diets hs been well defined (Knzw, 1985). The rtio of linolenic nd linoleic cids in prwn diets my lso be importnt. In the present study, the linolenic : linoleic rtio of 2-3 : 1 seemed to support the best prwn growth. The greter proportion of linoleic to linolenic cid s shown in the prwns fed on the commercil diet ws less effective in incresing the prwn s weight gin, suggesting tht the former is inferior to tht of the ltter FA. This finding grees well with Knzw et l. (1979) who noted tht both linolenic nd linoleic were essentil,but linolenic ws superior for stisfctory juvenile P. jponicus growth. When the proportion of linolenic cid ws reltive high (more thn 6 % of totl FA), however, the growth response of the prwns did not improve. The present study showed tht length ws significntly lower (P<.5) in the prwns fed on Ar-Dt which contined reltively greter proportion of 18:3n3 thn those prwns fed on Ar-Ti nd Ar-Ts. These dt suggest the excessive proportion of linolenic cid in the diet my not significntly increse the prwn weight gin s indicted erlier in nother peneid species (Rees et l., 1994; Merichn nd Shim, 1997). Regrdless of the FAs composition, erlier studies on the complete replcement of live feed by n rtificil diet reported slower growth rte in some peneid species lrve, lthough the survivl ws similr to the prwns fed on live feed (Jones et l., 1987; Kumrly et l., 1989). More studies hve confirmed tht the preference of live feed in lrvl diets ws relted to trypsin ctivity, the dominnt digestive enzyme during lrvl development (Le Vy et l., 1993). Artemi my provide redily or more esily digestible nutrients which improve the bility of prwns to meet their nutritionl requirements. T. Iso contined greter proportion of DHA compred to the other microlge used. However, when this microlge ws fed to Artemi, the content of DHA in Artemi ws reltively low, suggesting retroconversion of DHA to lower chin PUFA (Estevez et l., 1998) or this EFA is not incorported into the lipid of Artemi. Similr findings were reported by Volkmn et l. (1993). When microlge rich in DHA were fed to rotifers nd Artemi, they found the content of DHA in rotifers incresed, but not in Artemi. Nevertheless, in the present study, there ws n incresed in the proportions of DHA when Artemi were fed to the prwn. This FA, is possibly derived from 18:3n3 which ws bundnt in the Artemi. Severl studies hve shown the bility of lrvl nd juvenile P. jponicus to convert the linolenic cid, 18:3n3 into EFA, EPA nd DHA (Jones et l., 1979; Teshim et l., 1992). However, the conversion my be very slow, s indicted in the other Artemi fed prwns. The proportion of DHA ws less in the prwns fed on Ar-Ts nd the prwns fed on Ar-Dt thn in the initil prwn compred to the prwns fed on Ar-Ti. Kym et l. (198) reported the cpcity of bioconversion of linoleic nd linolenic cids to n-6 nd n-3 HUFA were less ctive in juvenile P. jponicus. The results of the present study suggest tht the contribution of 18:3n3 seems to be insufficient to supply DHA vi elongtion nd desturtion so tht supplementtion of dietry DHA is essentil in the prwn diet. This study found similr pttern of FA in unfed prwn nd the initil prwn, which my led to n ssumption tht the prwns hd ttcked nd eten the other prwns in the tnk to sustin life under strved conditions. High mortlity in the unfed tretments coupled with some growth in weight suggests cnniblism occurred in the present experiments. Some FA, such s 18:1n9, 18:1n7, 18:2n6, 18:3n3, 2:5n3 in unfed prwns were shown to follow the similr ptterns s those seen in the prwns t the beginning of the experiment. However, the DHA ws detected in low proportion in unfed prwn. 31 1

6 These results my be due to poor or slow incorportion of this FA tht ws obtined through cnniblism (s in Artemi, discussed bove). Another possibility ws tht the unfed prwns hve ctbolised this DHA to sustin their life during strvtion. Some crustcens hve been found to selectively ctbolise this EFA during strvtion (Olsen, 1998). Another FA tht my decrese during strvtion is plmitic cid (16: ) s occurred in the present study. This ftty cid my lso be utilised by the prwns under criticl condition, such s strvtion. Gury et l. (1975) in Bottino et l. (198) hs indicted the decreses of plmitic cid level in prwn lipids when prwns were left strved. IV. CONCLUSIONS There ws no signicnt difference in survivl between fed tretments. The FA composition of prwns, P. jponicus reflect their dietry lipid composition. The gretest weight gin rtes ppered in the prwns fed on Artemi rised on T. iso. And overll, the use of live feed produced greter response in terms of prwn weight gin compred to the commercil diet. The polyunsturted ftty cid of n3 series ppered to be more effective for supporting penned prwn growth. REFERENCES Bottino, N.R., gennity, J., Lilly, M.L., Simmons, E., Finne, G., 198. Sesonl nd nutritionl effects of the ftty cids of three species of shrimp, Peneus setiferus, P. ztecus nd P. duorrum. Aquculture 19, Brown, M. R., Jeffrey, S.W., Grlnd, C.D., Nutritionl spects of microlge used in mriculture; literture review. CSIRO Mrine Reserch Lbortories, Austrli. 44 pp. D Abrmo, L.R., Tricyglycerols nd ftty cids. In: D Abrmo, L.R., Conklin, D.E., Akiym, D.M. (Eds)., Crustcen nutrition. World Aqu. Soc. 6, Dintith, M.,1996. Rotifers nd tremi for mrine quculture : trining guide. An quculture sourcebook publiction in ssocition with the Ntionl Key centre for Aquculture, Univ. of Tsmni t Lunceston. Turtle Press Pty Ltd. 31 pp. D Souz, F.M.L., Lonergn, N.R., Effects of monospecific nd mixed lge diets on survivl, development nd ftty cid composition of peneid prwn (Peneus spp.) lrve. Mr. Biol. 133, Estevez, A., McEvoy, L.A., Bell, J.G., Srgent, J.R., Effects of temperture nd strvtion time on pttern nd rte of loss of essentil ftty cids in Artemi nuplii previously enriched using rchidonic cid nd eicospentenoic cidrich emulsions. Aquculture 165 (3-4), Jones, D.A., Kurmly, K., Arshrd, A., Peneid shrimp htchery trils using microencpsulted diets. Aquculture 64, Knzw, A., Nutrition of peneid prwns nd shrimps. In: Tki, Y., Primver, J.H., Lobrer, J.A. (Eds.), Proceedings of The First Interntionl Conference on the Culture of peneid prwns/shrimps. Iloilo city, Philippines, pp Knzw, A., Teshim, S., Tokiw, S., Ceccldi, H.J., Effects of dietry linoleic nd linolenic cids on growth of prwn. Ocenologic Act 2(1), Kym, M, Hirt,M., Knzw, A.,Tokiw, S.,Sito, M.,198. Essentil ftty cids in the diet of prwn-iii.lipid metbolism nd ftty cid composition.bull. Jpn. Soc. Sci.Fish. 46(4), Kumrly, K., Jones, D.A., Yule, A.B., Est, J., Comprtive nlysis of the growth nd survivl of P. monodon (Fbricius) lrve, from protozoez 1 to postlrve 1, on live feeds, rtificil diets nd combintions of both. Aquculture 81, Le Vy, L., Rodriguez, A., kmruddin, M.S., Jones, D.A., Influence of live nd rtificil diets on tissue composition nd trypsin ctivity in P. jponicus lrve. Aquculture 118,

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