Kamil Mert Eryalcin 1, * Introduction

Transcription

1 Turkish Journl of Fisheries nd Aqutic Sciences 18: (2018) ISSN DOI: / v18_1_09 RESEARCH PAPER Effects of Different Commercil Feeds nd Enrichments on Biochemicl Composition nd Ftty Acid Profile of Rotifer (Brchionus Plictilis, Müller 1786) nd Artemi Frnciscn Kmil Mert Erylcin 1, * 1 Istnul University, Fculty of Fisheries, Aquculture Deprtment, Phytoplnkton nd Zooplnkton Culture Lortory, Ordu Cd. No: 200, Lleli, 34470, Istnul, Turkey. * Corresponding Author: Tel.: /16462; Fx: ; E-mil: erylcin@istnul.edu.tr Astrct Received 28 Septemer 2016 Accepted 15 My 2016 The ojective of the present study ws to determine the effects of severl commercil rotifer feeds nd enrichments on growth, iochemicl nd ftty cid composition of L-type rotifer nd Artemi frnciscn nuplii. In experiment I, five experimentl diets (M0 Plus, S.Prkle, w-3 Yest 60, Beker s yest nd Chlorell vulgris) were tested for rotifer culture performnce nd in the second experiment six enrichers (w-3 Olio, n-3 Top Rich, Red Pepper, Culture Selco, microlge mixture (Dunliell slin + Chlorell vulgris) nd Emulsion T) were evluted for the ftty cid composition of rotifer nd Artemi frnciscn. In experiment I, rotifers fed S.Prkle nd Beker s yest showed etter iomss production nd egg density while the numer of egg crrying femle numer ws higher. In experiment II, rotifers enriched n-3 Top Rich showed etter n-3 HUFA retention wheres Artemi frnciscn nuplii enriched Red Pepper showed highest HUFA ccumultion. Culture Selco seems optiml for rtemi enrichment for EPA (20:5n-3) nd DHA (22:6n-3) ccumultion. In conclusion, Beker s yest is still pplicle in comprison to other commercil feeds in rotifer culture. Further study is needed for determintion of minerl composition of rotifer feeds nd commercil enrichments nd their retention in live prey nd lrve use in Turkey. Keywords: Rotifer, rtemi, growth, enrichment, ftty cids. Introduction Successful production of mrine fish lrve is limited y the poor chivement of htching rte nd lck of essentil nutrients in live prey nd microdiets (Izquierdo & Koven, 2011). Live prey re necessry in order to stimulte digestive enzymes nd sustin enough energy from exogenous feeds (Knzw, 2003; Hmre, Yufer, Rønnestd, Boglione & Conceiço & Izquierdo, 2013). Therefore, utiliztion of rotifers nd Artemi hs high importnce t lrvl stge, since feeding with dry diets delys gut development t tht stge (Srgent, McEvoy, Estevez, Bell, Bell, Henderson & Tocher, 1999; Sorgeloos, Dhert, & Cndrev, 2001; Hmre, Srivstv, Rønnestd, Mngor-Jensen & Stoss, 2008). However, rotifer nd rtemi re known to lck essentil nutrients such s polyunsturted ftty cids (PUFA), essentil mino cids (EAA), vitmins nd minerls (Hmre, Srivstv, Rønnestd, Mngor-Jensen & Stoss, 2008; Hmre, 2011). For tht reson, live prey should e enriched with essentil nutrients efore they re given to fish lrve (Ferreir, Msed, Fáregs, & Otero, 2008; Cstillo, Gpsin, & Leño, 2009; Demir & Diken, 2011; Mhre, Hmre, & Elvevoll, 2012). Nutritionl composition of live prey is differed mong htcheries nd this is n ctul topic of recent reserches (Hmre, 2016). The highly unsturted ftty cids (HUFA) such s docoshexenoic (DHA;22:6n-3), eicospentenoic cid (EPA; 20:5n-3) nd rchidonic cid (ARA; 20:4n-6) hve importnt functions in mrine fish nd crustcen metolism (Izquierdo, Socorro, Arntzmendi, & Hernández-Cruz, 2000; Izquierdo, 2005). Another essentil nutrient, minerls re ecoming more importnt in enrichment nd feeding process of live prey. Most populr rotifer feed, eker s yest hs negtive effect on nutritionl vlue of rotifers (Lie, Hlnd, Hemre, Mge, Lied, Rosenlund, Sndnes, & Olsen, 1997; Hmre, 2016) wheres freshly cultured microlge re common enricher to otin good nutritionl profile in htcheries. Due to difficulties in mnipultion nd culture of microlge, commercil products re widely used for feed nd enrichment process. For instnce, enrichment with Spirulin incresed mino cid levels in rtemi (Bhvn, Devi, Shnti, Rdhkrishnn, & Poongodi, 2010) y which Pulished y Centrl Fisheries Reserch Institute (CFRI) Trzon, Turkey in coopertion with Jpn Interntionl Coopertion Agency (JICA), Jpn

2 82 K. M. Erylcin / Turk. J. Fish. Aqut. Sci. 18: (2018) importnt mino cids such s turine could e trnsferred to lrve. Turine is known s free orgnic cid tht plys importnt role in lipid digestion, sorption nd osmoregultion (Kim, Mtsunri, Tkeuchi, Yokoym, Murt, & Ishihr, 2007; Tkeuchi, 2014). The turine concentrtion of rotifer nd rtemi re lower thn tht of copepods (Vn der Meeren, Olsen, Hmre, & Fyhn, 2008). Therefore, one of the experimentl group ws designed s Turine source. Severl rotifer diets nd enrichments re ville on the mrket with different nutritionl compositions. The products tested in our experimentl groups re commonly used for enrichment process of live prey in Turkey. However, the effects of those products on ftty cid composition were not yet reported in detils. The im of the present study ws to determine the effects of commonly used rotifer diets on rotifer culture performnce (Experiment I); M0 Plus, S.prkle, w-3 yest 60, Beker s yest nd Chlorell vulgris nd the effects of severl enrichments; Olio w-3, Red Pepper, Top Rich, Culture Selco, mixture of freshly cultured microlge species (Dunliell slin nd Chlorell vulgris) nd Turine emulsion (Emulsion T) (Experiment II) were evluted on growth, iochemicl nd ftty cid compositions of rotifer nd rtemi. Mterils nd Methods Microlge Culture The originl strin of Dunliell slin nd Chlorell vulgris were provided y the Culture Collection of Alge nd Protozo (CCAP) culture collection. Microlge culture were strted in test tues (30 ml.) kept under 22 o C t 40 µe m -2 s -1 light intensity, 12/12 light-drk photoperiod for 25 dys. Culture volume ws incresed to test tues to 5 lt. Erlenmeyer s flsks t density of cell ml -1. Culture flsks contining 2 lt. sterilized f/2 medium inoculted with 2 test tue stock cultures of Dunliell slin t egining. Growth of microlge culture ws determined y using Neueur counting chmer ech dy. All cultures were crried out in triplictes nd mens re presented. 3N-BBM+V medium ws used for Chlorell vulgris culture descried y CCAP (Scotlnd, UK). homogenized with 40 ml. of this solution for 3 minutes using four-lde lender t rpm. Resulting emulsions were immeditely used for the following enrichment. Rotifer Stock Culture Rotifers (Brchionus plictilis, L-strin, men loric length µm) were otined from Kılıç Se Products Inc., Güvercinlik Htchery, Bodrum, Muğl, Turkey. Rotifers were cultured in two 135-L conicl tnks t room temperture under continuous illumintion during 24 hours t lortory. Stock rotifers were fed dily t 9:00.m. with Nnnochloropsis gditn t cells/ml. concentrtion nd 04:00 p.m. t level of 0.03 g L -1 instnt Bker s yest (Scchromyces cerevisie). Rotifers were cultivted in semi-continuous cultures. The wter temperture (21.2 ± 0.5 C) nd slinity (% o 25) were mesured dily during cultivtion of stock culture. Rotifers were strved for one 12 hours nd wshed with sterilized sewter efore experiments strted. Experiment I: Rotifer Feeding Experiment At the eginning of the rotifer feeding experiment, when rotifer concentrtion reched 350 rotifer/ml in circulr flt ottom 135 liters fierglss stock tnk, they were distriuted into fifteen experimentl conicl glss flsks (10 lt.) reprensent five experimentl groups in triplicte. Five experimentl rotifer feeds were evluted for 8 dys feeding. The tretments for first experiment were 1) M0 Plus (0.40 g for 10 6 ind. ml -1 ), 2) S.prkle (0.65 g for 10 6 ind. ml -1 ), 3) w-3 yest 60 (0.50 g for 10 6 ind. ml -1 ), 4) Beker s yest (0.5 g for 10 6 ind. ml -1 ) nd 5) Chlorell vulgris. Ech experimentl feed ws tested in triplicte nd pplied t dily rtion of 0.8 g dry weight (DW) 10 6 rotifer. dy -1. During the experiment, egg numer, juvenile numer, egg crrying femle numer nd totl rotifer iomss were counted nd clculted every dy. The smpling consisted of tking 3 times one milliliter smple collected with glss pipette from ech tretment nd dd Lugol solution in order to settle rotifers for counting. Experiment II: Rotifer nd Artemi Enrichment Preprtion of Emulsion T Type-A geltin (from porcine skin, loom; Sigm, St. Louis, MO) ws dissolved in wter t 50 C to produce 1% (wt/wt) solution (Ando, Ae, Ookuo, & Nmikw, 2005). After the solution ws cooled to room temperture, fish oil (Cpelin oil, Denof, Fredrikstd, Norwy; 3 g), Turine (Turine Acros, New Jersey, USA; 1 g), vitmin B 1 nd B 12 (Sigm Aldrichi, Steinheim, Germny; 0.01 g) were Rotifer Enrichment Thirty million rotifers were hrvested from the stock culture tnk (stock density ws 400 rotifer ml -1 ) for enrichment experiment. Initil rotifer density ws 60 rotifer ml -1 in ech tretment prllels. Rotifers were enriched with five enrichment products twice dy t 7.m. nd 7 p.m. sed on recommendtion y products mnufcturer nd two of them were prepred in lortory s microlge mixture nd Emulsion T

3 K. M. Erylcin / Turk. J. Fish. Aqut. Sci. 18: (2018) 83 (lipid nd Turine mixture). Amount of enrichment products were clculted ccording to rotifers density in ech experimentl 1000 ml flsk nd ll enrichers were homogenized using hnd lender for 2 minutes. Clcultion for ech enrichers ws s follows; ) 600 mg L -1 of Olio w-3 (Bernqu, Belgium) ) 180 mg L -1 of Red Pepper pste (Bernqu, Belgium) c) 150 mg L -1 of (n-3) Top Rich (Rich S.A., Greece) d) mix of cells/ml. Chlorell vulgris nd cells/ml. Dunliell slin e) 0.8 g L -1 of Emulsion T. Rotifer iomss (25 grm) were hrvested on 75 µm plnkton nets fter 24 hours, rinsed with sterilized sewter for 3 minutes, then stored in -80 C for iochemicl nd ftty cid nlysis. Ech tretment ws crried out in triplicte. Artemi Enrichment nd identified y comprison with externl stndrds nd well chrcterized fish oils (EPA 28, Nippi, Ltd Tokyo, Jpn). Sttisticl Anlysis All dt were sttisticlly treted using SPSS Sttisticl Softwre System 15.0 (SPSS, The significnt level for ll the nlysis ws set 5%, nd results re given s men vlues nd results stndrd devition. Also, ll vriles were checked for normlity nd homogeneity of vrince, using the Levene s tests respectively. To compre mens, the group dt were sttisticlly tested using one-wy ANOVA. When vrinces were not homogente, non-prmetric Kruskl Wllis test ws ccomplished. Artemi frnciscn metnuplii were otined from the htching of Gret Slt Lke Artemi cysts (INVE Aquculture Nutrition, Dendermonde, Belgium). Cysts were decpsulted s descried y Bruggemn et l. (1980), then incuted during 24 h in 5 L cylindricl-conicl glss tnk contining sewter (38 g L -1 slinity) t 28 C. Gentle ertion ws plced in ottom of the glss tnk with continuous illumintion. After htching, Instr I nuplii were moved into cylindricl continers t 23 o C nd rinsed with UV-treted sewter for 20 minutes. Artemi enrichment ws conducted in 10 L conicl glss continer t density of individuls L -1 density t 22 C with moderte ertion. Six enrichment procedures were evluted nd rtemi nuplii groups were enriched twice dy t 7.m. nd 7 p.m. with following procedures; ) 250 mg L -1 of Olio w-3 (Bernqu, Belgium) ) 750 mg L -1 Red Pepper pste (Bernqu, Belgium) c) 300 mg L - 1 of (n-3) Top Rich (Rich S.A., Greece) d) 0.1 g L -1 Culture Selco (INVE Aquculture, Belgium) e) mix of Chlorell vulgris nd Dunliell slin concentrtion of 3:1, respectively (from stock concentrtion , cell/ml.) nd f) 0.8 g L -1 Emulsion T. All enriched rtemi were hrvested (40 g from per tretment) fter 24 h on 125 m filter nd rinsed with utoclved sewter for 5 min then stored in -80 C for iochemicl nd ftty cid nlysis. Ech tretment ws crried out in triplicte. Proximte nd Ftty Acid Composition Moisture (A.O.A.C., 1995), protein (A.O.A.C., 1995) nd crude lipid (Folch, Lees, & Stnley, 1957) contents of rotifer nd rtemi were nlyzed. Ftty cid methyl esters were otined y trnsmethyltion with 1% sulphuric cid in methnol (Christie, 1989). Ftty cid methyl esters were seprted y GC (GC- 14A; Shimdzu, Tokyo, Jpn) using helium s crrier gs. Ftty cid methyl esters were quntified y FID following the conditions (Izquierdo, Wtne, Tkeuchi, Arkw, & Kitjim. 1990) Results Rotifer Growth Performnce Egg numer ws found higher in rotifers fed with the S.prkle t dy 3 (Figure 1). After tht dy, egg numer strted to decrese in feeding experiment. At finl dy of the experiment, S.prkle nd Chlorell vulgris groups showed highest numer of egg. Juvenile rotifer numer ws found higher in Beker s yest group then tht in other groups (Figure 1). Among the groups, S.prkle nd Beker s yest showed the est egg production numer. Egg crrying femle numer ws decresed strting from feeding of dy 2 (Figure 2c). Totl iomss of rotifer ws enhnced y feeding with S.prkle nd Beker s yest t culture of dy 6 (Figure 2d). Biochemicl Composition of Rotifer nd Artemi At the end of the study, crude protein content of rotifer ws different mong groups (P<0.05). Microlge mixture enriched rotifers showed higher protein content thn tht in other groups. Lipid content ws significntly higher in rotifer-enriched n- 3 Top Rich group (P<0.05). Ash content ws higher in Olio w-3 group rotifers nd the lowest vlue ws in rotifer enriched Emulsion T groups (P<0.05). Similrly, moisture content of rotifer enriched Emulsion T ws found higher thn Microlge Mix, Red Pepper nd Olio w-3 nd n-3 Top Rich, respectively (P<0.05) (Tle 1). Crude protein content of rtemi ws not significntly (P>0.05) different mong experimentl groups. However, lipid content of rtemi enriched Red Pepper showed higher vlue thn other groups (P<0.05). Ash content of rtemi ws significntly higher in Emulsion T group thn tht in other groups (P<0.05). Moisture contents were significntly different mong groups nd the highest level ws in W3 Olio group rtemi (P<0.05) (Tle 2).

4 J u v e n ile (in d m l -1 ) E g g d e n s ity (e g g s m l -1 ) 84 K. M. Erylcin / Turk. J. Fish. Aqut. Sci. 18: (2018) 8 0 M 0 P lu s 6 0 S P r k le w 3 Y e s t 6 0 B eker's yest 4 0 C h lorell vulgris D y s M 0 P lu s S p r k le w 3 Y e s t 6 0 B e k e r's y e s t C h lorell vulgris D y s Figure 1. Egg () nd juvenile numer () of rotifer fed with different commercil feeds nd microlge (men±sd, n=5). Vlues with the sme superscript re not significntly different (P<0.05). Ftty Acid Composition of Rotifer nd Artemi At the end of the enrichment, EPA (20:5n-3), DHA (22:6n-3), totl n-3, totl n-6, totl n-3 HUFA levels of rotifer were enhnced y utiliztion of n-3 Top Rich product (P<0.05). Similrly, linoleic cid (LA; 18:2n-6) nd linolenic cid (LNA; 18:3n-3) levels were higher in rotifer-enriched n-3 Top Rich. n- 3/n-6 rtio ws lso higher in n-3 Top Rich group. However, ARA (20:4n-6) level of rotifers ws supported y utiliztion of microlge mixture (P<0.05). Oleic cid (18:1n-9) level ws higher in rotifer enriched Emulsion T (Tle 3). Similrly, oleic cid level ws supported in rtemi enriched Emulsion T nd Microlge mixture groups (P<0.05). Totl n-3 HUFA level ws higher in Red pepper group wheres EPA, DHA, totl n-3 nd totl n-6 levels were higher in rtemi enriched Culture Selco. ARA level of rtemi ws high in red pepper group (P<0.05) (Tle 4). Discussion Live prey re much lower in certin essentil ftty cids, mino cids, vitmins nd minerls thn copepods which re the nturl prey of mrine fish lrve (Luzens, Zmor, & Brr, 2001; Hmre, Srivstv, Rønnestd, Mngor-Jensen, & Stoss, 2008; Hmre, 2016). Therefore, in order to get high survivl in mrine fish lrve, higher nutrient qulity nd quntity in rotifer nd rtemi should e mintined t lrvl period (Izquierdo & Koven, 2011). Enrichment process is one of the most importnt ppliction in mrine fish htcheries. Enrichment products nd methods (intervls nd ppliction time) ply importnt role for the retention of essentil nutrients in live prey nd protocols differ mong htcheries (Furuit, Tkeuchi, Toyot, & Wtne, 1996). Despite the importnce of nutritionl vlue of live prey in development nd survivl of mrine fish lrve, the nutritionl effects of different feeds nd enrichment products re still not optimized (Izquierdo, 1996; Izquierdo, Socorro, Arntzmendi, & Hernández-Cruz, 2000; Monroig, Nvrro, Amt, Gonzlez, Bermejo, & Hontori, 2006). Therefore, in this study, the effects of different rotifer feeds nd enrichments on growth, iochemicl composition nd ftty cid profile of rotifer-rtemi were investigted. Rotifer numers need to e incresed rpidly

5 R o tife r B io m s s (in d L - 1 ) F e m le w ith E g g (in d m l -1 ) K. M. Erylcin / Turk. J. Fish. Aqut. Sci. 18: (2018) M 0 P lu s S p r k le 4 0 w 3 Y e s t 6 0 B e k e r's y e s t C h lorell vulgris 2 0 c D y s M 0 P lu s S p rk le w 3 Y e s t 6 0 B eker's yest C h lo re ll v u lg ris d D y s Figure 2. Egg crried femle (c) nd totl rotifer iomss (d) numer during feeding experiment. (men±sd, n=5). Vlues with the sme superscript re not significntly different (P<0.05). Tle 1. Proximte composition (g 100 g -1 dry weight) for whole ody of rotifer iomss t the end of the enrichment experiment. Different letters within line denote significnt differences (P<0.05). Vlues expressed in men ± SD (n =3 tnks/diet) Rotifer Control n-3 Top Rich Olio w3 Red Pepper Microlge Mix Emulsion T Crude protein 52.59± ±0.65 d 57.14±0.85 c 59.05± ± ±0.96 Crude lipids 8.96± ± ±0.58 c 13.25± ±0.32 c 10.52±0.89 c Ash 1.62± ± ± ±0.22 c 1.11± ±0.44 d Moisture 9.65± ±0.24 d 8.85±0.45 e 9.70±0.85 c 10.22± ±0.65 Tle 2. Proximte composition (g 100 g -1 dry weight) for whole ody of rtemi t the end of the enrichment experiment. Different letters within line denote significnt differences (P <0.05). Vlues expressed in men ± SD (n =3 tnks/diet) Artemi Control n-3 TOP Microlge W3 Olio Red Pepper RICH Mix Emulsion T Selco Crude Protein 4.67± ± ± ± ± ± ±0.65 Crude Lipid 0.45± ±0.01 d 0.79±0.02 cd 1.31± ± ±0.04 c 0.72±0.04 d Crude Ash 0.72± ± ± ± ±0.02 c 0.34± ±0.32 c Moisture 6.05± ± ± ± ±1.22 c 7.98± ±0.26 during fish lrvl production period. At this point, physicl, chemicl nd nutritionl fctors ply importnt role (Luzens, Zmor, & Brr, 2001). Nowdys, not only commercil rotifer feeds ut lso freshly cultured microlge re eing evluted for rotifer cultivtion. Özş, Göksn, & Ak (2006) hve reported tht totl rotifer nd egg crrying femle rotifer numers were continuously incresed y feeding fresh microlge (Nnnochloropsis sp.). Similrly, Nhu (2004) hs mentioned out the eneficil effects of fresh Nnnochloropsis ocult on mximum growth rte of rotifer. In nother study, freshly cultured Pvlov viridis significntly incresed rotifer nd egg numers wheres egg rtio ws enhnced y Beker s yest fed rotifers in lortory scle experiment (Reherg-Hs, Meyer,

6 86 K. M. Erylcin / Turk. J. Fish. Aqut. Sci. 18: (2018) Tlo 3. The ftty cid compositions of rotifers (Brchionus plictilis) enriched twice dy nd fter 24h of enrichments (% totl ftty cids) Unenriched n-3 TOP Microlge Olio w3 Red Pepper RICH Mix Emulsion T 4: : : c d 13: : d d c 14: : c d d 16: d c c 16: c : : c :1n d c e :2n e c d :3n d c d :3n : :1n d c :3n c :3n c 20:4n c c 20:5n c 22: : :5n :6n c 23: : : Σ Sturted d c e Σ Monounsturted c Σ n c c Σ n e c d Σ n d c e Σ n-3 HUFA EPA/ARA DHA/EPA DHA/ARA n-3/n c c c Lippemeier, & Schulz, 2015). At the sme study, lrge scle rotifer cultivtion ws investigted y severl microlge feeds nd Nnnochloropsis sp. utiliztion resulted etter rotifer culture performnce (Reherg-Hs, Meyer, Lippemeier, & Schulz, 2015). Chlorell vulgris hs een noted s suitle feed for Prole similis (Lee, Kim, Lee, Hgiwr, Kwon, Prk, & Prk, 2016). Despite Chlorell vulgris hs higher iomss potentil, protein nd B 12 content in comprison to Nnnochloropsis ocult (Mruym, Nko, Shigeno, Ando, & Hirym, 1997), the utiliztion of freshly cultured Chlorell vulgris did not well perform in lortory scle rotifer culture in our study. In fct, the highest totl rotifer numer ws incresed y feeding Beker s yest nd S.prkle products. In greement with our findings, Lind (2014) reported the highest rotifer growth in those fed S.prkle nd Reed Rotigrow Nnnochloropsis. Moreover, S.prkle sustitution y Beker s yest with different proportions hs not resulted in significnt differences in terms of growth. In our study, one of the reson of insignificnt effect of Chlorell vulgris on rotifer growth performnce could e relted to low level of B 12 s Hirym, Mruym, & Med (1989) reported. However, Virio concentrtion incresed when rotifer fed 75% Beker s yest sustituted diet. These results could e relted to vitmin B 12 nd protein content of Scchromyces cerevisie s previously reported in other studies (Luzens, Tndler, & Minkoff, 1989; Ferreir, Pinho, Vieir, & Tvrel, 2010). Conversely, different forms of yest hve positive

7 K. M. Erylcin / Turk. J. Fish. Aqut. Sci. 18: (2018) 87 Tle 4. The ftty cid compositions of rtemi enriched twice dy nd fter 24h of enrichments (% totl ftty cids) Unenriched n-3 TOP Microlge W3 Olio Red Pepper RICH Mix Emulsion T Selco 4: : : d d e c 13: : e c e d 14: : c d cd 16: c c c d 16: c : c 18: e c d f 18:1n c :2n d c c e :3n e c d f :3n : c c : :1n c :3n :3n :4n c :5n : : c c : :5n :6n : : : Σ Sturted d c e Σ Monounst d c c c urted Σ n e c d Σ n e c d f Σ n d c c Σ n-3 HUFA c EPA/ARA DHA/EPA DHA/AR A c n-3/n d c effect on rotifer growth. For instnce, Nhu (2004) lso otined mximum density of rotifer when fed with wet yest feed. Not only vitmins ut lso other micronutrients re lso importnt for rotifer culture performnce (Yoshimtsu, Higuchi, Zhng, Fortes, Tnk, & Yoshimur, 2006). Ftty cid composition of rotifer nd rtemi were improved y enrichment products in our study. As primry energy source, oleic cid level of rotifers ws fortified y Emulsion T enrichment in Experiment II. In fct, oleic cid is primer energy source for fish lrve nd it is very importnt ftty cid for lrvl energy udget especilly t first exogenous feeding period (Chu & Infnte, 2001; Yufer & Dris, 2007). Therefore, content of higher oleic cid level is desirle in live prey (Izquierdo, 1996). Totl n-3 level of rotifer ws supported y n-3 Top Rich product. However, ARA level ws significntly higher in rotifer enriched microlge mixture. Therefore, the comined usge of couple of enrichers in live prey seem to e good option in order to mintin optimum essentil ftty cid rtio. Bsed on our results, n-3 Top Rich nd freshly cultured microlge mixture is highly

8 88 K. M. Erylcin / Turk. J. Fish. Aqut. Sci. 18: (2018) recommendle for rotifer enrichment. As for rtemi, enrichment with Culture Selco seemed to e the est option for improving the nutritionl vlue of rtemi wheres ARA level ws enhnced y the utiliztion of Red Pepper. ARA is very importnt ftty cid tht tkes ction in survivl, immune metolism nd stress resistnce in mrine fish (Bell & Srgent, 2003). Hn, Geurden, & Sorgeloos (2001) hve rgued tht when rtemi enriched emulsions contining essentil ftty cids with different proportions such s DHA/Oleic cid, EPA/Oleic cid nd AA/Oleic cid resulted in different retention of ftty cids. Among them EPA nd oleic cid were the most efficiently remined ftty cids in rtemi t the end of the 24 hours enrichment. To sum up, the comined usge of more thn one enrichment for rtemi could e good the est option, such s Red Pepper nd Culture Selco. In this study, we specificlly focused on culture performnce of rotifer nd ftty cids profile of enriched rotifer nd rtemi. Bker s yest Scchromyces cerevisie is still chep feed source nd seemed to e performed efficiently in rotifer culture. Nevertheless, further experimentl studies on feeding y lterntive yest products in rotifer cultivtion re necessry. In terms of enrichment process, EPA nd DHA levels were supported y n-3 Top Rich product in rotifer. However, ARA level of rotifer ws incresed y the utiliztion of fresh microlge mixture. Culture Selco is strongly dviced for nutritionl oosting of rtemi. The eneficil support of the commercil products in terms of ftty cid composition nd vitmins (C nd E) ws lso suggested in Hmre (2016). Recently, micronutrients such s iodine, selenium, copper nd mngnese hve ecome more importnt for enrichment process (Penglse, Hmre, Sweetmn, & Nordgreen, 2011; Rieiro, Rieiro, Dinis, & Moren, 2011; Srivstv, Hmre, Stoss, & Nordgreen, 2012; Nordgreen, Penglse, & Hmre, 2013). Delivering micronutrients to rotifers cn e mnged y enriching microlge (Douch, Livnsky, Kotrcek, & Zchleder, 2009; Kou, Velíšek, Strá, Msojídek, & Kozák, 2014). This could e n edequte solution if nutritionl vlue of microlge could e incresed. Further studies re needed for investigtion into minerl composition in enrichments nd retention in live prey in feeding nd enrichment of live prey in Turkey. Acknowledgements This study ws supported y Istnul University Scientific Reserch Projects (BAP Project No: 28086). References A.O.A.C. (1995). Officil Methods of Anlysis of Officil Anlyticl Chemists Interntionl 16th (Eds.), Assocition of Officil Anlyticl Chemists, Arlington, VA, USA. Ando, Y., Ae, T., Ookuo, Y., & Nmikw, S. (2005). Ftty Acid Enrichment of Rotifers Using Type-A Geltin Solution. 北海道大学水産科学研究彙報, Bulletin of Fisheries Sciences, Hokkido University, 56(3), Bell, J. G., & Srgent, J. R. (2003). Archidonic cid in quculture feeds: current sttus nd future opportunities. Aquculture, 218(1), Bhvn, P.S., Devi, V.G., Shnti, R., Rdhkrishnn, S., & Poongodi, R. (2010). Bsic iochemicl constituents nd profiles of mino cids in the post lrve of Mcrorchium rosenergii fed with Spirulin nd Yest enriched Artemi. Journl of Scientific Reserch, 2(3), Bruggemn, E.; Sorgeloos, P.; Vnhecke, P. (1980). Improvements in the decpsultion technique of Artemi cysts, in: Persoone, G. et l. (Ed.) The rine shrimp Artemi : Proceedings of the Interntionl Symposium on the rine shrimp Artemi slin, Corpus Christi, Texs, USA, August 20-23, 1979: 3. Ecology, culturing, use in quculture. pp Chu, C., & Infnte, J.Z. (2001). Sustitution of live food y formulted diets in mrine fish lrve. Aquculture, 200(1), Cstillo, E.C., Gpsin, R.S., & Leño, E.M. (2009). Enrichment potentil of HUFA-rich thrustochytrid Schizochytrium mngrovei for the rotifer Brchionus plictilis. Aquculture, 293(1), Christie, W.W. (1989). Gs Chromtogrphy nd Lipids: Prcticl Guide. The Oily Press, Bridgwter, U.K pp. Demir, O., & Diken, G. (2011). Effects of commercil enrichment products on ftty cid components of rotifer, Brchionus plictilis. Africn Journl of Biotechnology, 10(66), Douch, J., Livnsky, K., Kotrcek, V., & Zchleder V. (2009). Production of Chlorell iomss enriched y selenium nd its use in niml nutrition: review. Applied Microiology Biotechnology, 83, Ferreir, M., Msed, A., Fáregs, J., & Otero, A. (2008). Enriching rotifers with premium microlge. Isochrysis ff. gln clone T- ISO. Aquculture, 279(1), Ferreir, I.M.P.L.V.O., Pinho, O., Vieir, E., & Tvrel, J.G. (2010). Brewer's Scchromyces yest iomss: chrcteristics nd potentil pplictions. Trends in food science & Technology, 21(2): Folch, J., Lees, M., & Stnley, G.H.S. (1957). A simple method for the isoltion nd purifiction of totl lipids from niml tissues. Journl of Biology Chemistry, 226, Furuit, H., Tkeuchi, T., Toyot, M., & Wtne, T. (1996). EPA nd DHA requirements in erly juvenile red se rem using HUFA enriched Artemi Nuplii. Fisheries Science, 62(2), Hmre, K., Srivstv, A., Rønnestd, I., Mngor Jensen, A., & Stoss, J. (2008). Severl micronutrients in the

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