Production of artificial diets for female broodstock of western white shrimp (Litopenaeus vannamei) and study on their singular effect

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1 Irnin Journl of Fisheries Sciences 16(4) Production of rtificil diets for femle broodstock of western white shrimp (Litopeneus vnnmei) nd study on their singulr effect Ghorbni Vghei R. *1 ; Abolhsni M.H. 2 ; Mtinfr A. 3 ; Ddgr Sh. 3 ; Ghorbni R. 4 Received: Jnury 2016 Accepted: August 2016 Abstrct Advntges of pelleted feeds in terms of storge, trnsporttion nd lower costs compred to nturl fresh feed justified the replcement of rtificil feeds insted of nturl ones prtilly on mturtion performnces. This study comprised nine tretments (with three replictions) including control tretment: polychete worm (Perinereis nunti), Squid nd clf liver (nturl feed); T 1 : pellet feed with 50% crude protein nd 8% crude lipid+ P. nunti, Squid nd clf liver; T 2 : pellet feed with 50% crude protein nd 10% crude lipid+ P. nunti, Squid nd clf liver; T 3 : pellet feed with 40% crude protein nd 10% crude lipid+ P. nunti, Squid nd clf liver; T 4 : pellet feed with 40% crude protein nd 8% crude lipid+ P. nunti, Squid nd clf liver P. nunti; T 5 : pellet feed with 50% crude protein nd 10% crude lipid; T 6 : pellet feed with 50% crude protein nd 8% crude lipid; T 7 : pellet feed with 40% crude protein nd 10% crude lipid; T 8 : pellet feed with 40% crude protein nd 8% crude lipid. In tretments 1, 2, 3 nd 4, 50% of the nturl diets were removed nd pellet foods used insted. The mount of pellet feeds tht given dily, ws 3.5% of shrimp broodstock biomss. In tretments 5, 6, 7 nd 8, the mount of pellet feeds tht given dily ws 7.5% dily. Feeding ws done 4 times dy. Gondosomtic index (%GSI) in the control (3.23%), tretment 3 (3.20%) nd tretment 6 (3.02%) were significntly higher thn tht in the other tretments (p<0.05). Absolute fecundity in the control (29980 eggs) nd tretment 3 (29683 eggs) ws significntly higher thn other tretments (p<0.05). Heptosomtic index (%HSI) reched the lowest level in tretment 8 (2.14%). But in tretments 3 nd 5 ws more thn control, but did not ny sttisticl significnt (p>0.05). Survivl rte in the tretment 8 (26.66%) ws significntly lower thn other tretments (p<0.05). Generlly, In the tretment 3, with the elimintion of 50% of the nturl foods, nd the use of pellet food insted of them, result of comprtive indices of reproduction were in desirle proportions. Keywords: Nturl wet feed, Pellet feed, Sexul mturtion, Western white shrimp femle brood 1- Shrimp Reserch Center, Irnin Fisheries Science Reserch Institute (IFSRI), Agriculturl Reserch Eduction nd Extension Orgniztion (AREEO), Bushehr, Irn 2- Deprtment of Nturl Resources, Fculty of Agriculture, Islmic Azd University, Khorsgn Isfhn Brnch, Esfhn, Irn. 3- Irnin Fisheries Science Reserch Institute (IFSRI), Agriculturl Reserch Eduction nd Extension Orgniztion (AREEO), Bushehr, Irn 4- Deprtment of Nturl Resources, Agriculture nd Nturl Resources, Gorgn, Irn * Corresponding uthor s E-mil: ghorbni_v2@yhoo.com

2 1205 Ghorbni Vghei et l., Production of rtificil diets for femle breeders of western white Introduction It hs been reported tht totl replcement of nturl feeds with rtificil ones hs not been chieved, yet (FAO, 2010). Severl fctors hve mde it impossible to use lrge mount of pellet feed for shrimp broodstock mturtion. Chimsung (2014) hve reported tht polychete worm improves shrimp reproduction due to hving high nutritionl vlue, rchidonic cid nd importnt reproduction hormones. In ddition, it hs been believed tht polychete worm is n excellent source for not only unsturted ftty cids, but lso sexul hormones similr to those of shrimp. Also, squid not only is rich source of cholesterol, but lso contins sexul steroids involved in vitellogenesis (Wouters et l., 2005). As mentioned before, nturl feeds for breeder feeding hve high mounts of n-3 ftty cid. Also, fecundity rte of broodstock is correlted with n-3 ftty cid (Wouters et l., 2005). Amino cids, ftty cids, protein nd lipid ply n importnt role in shrimp breeder mturtion nd need more protein during the sexul mturtion period (Coutteu, 2001). In ddition, highly unsturted ftty cids (HUFA) especilly eicospentenoic cid nd docospentenoic cid re importnt components of breeder diets nd re found undntly in ovry tissue (Teikw nd Mgy, 2003). Diets deficient in n-3 HUFA dversely ffect ovry development, fecundity nd egg qulity (Teikw nd Mgy, 2003). Western white shrimp broodstock with frequent spwning hve higher protein content in their heptopncres nd ovries thn those broodstock with wek spwning (Mglhes et l., 2012). Successful sexul mturtion in western white shrimp cn be obtined with freshly-frozen wet diet (with or without rtificil diets). The reserchers hve emphsized tht the weight of mtured shrimp ovries my increse 4-6 fold within out one week (Verstrete et l., 1995). Divn et l. (2009) reported tht pelleted feeds contining 50% protein nd 10% lipid cn be used in combintion with fresh feeds. Alv et l. (1993) reported tht diets lcking HUFA cuse decrese in gondo-somtic index (GSI). Consequently, it is believed tht HUFAs re needed for sexul mturtion. Pelleted feeds re importnt in mny spects, like ese of storge, trnsporttion, feeding nd low costs. The im of the present study ws to provide suitle pelleted diet for the sexull mturtion of femle broodstock nd determining their effects on sexull prmeters. Mterils nd methods Broodstock cclimtion A totl number of 270 femle broodstock with n verge weight of 37±2 g were smpled from Bndrgh Reserch Sttion nd kept in two 4-ton fiberglss tnks for one week to cclimtize prior to stocking in 300-L tnks. Eyestlk ltion ws done week fter stocking in the 300-L tnks

3 Irnin Journl of Fisheries Sciences 16(4) filled with 150 L wter. Feeding nd sexul mturtion in broodstock were checked every dy for 5 weeks. Tretments This study ws conducted with 9 tretments ech with three replictions contining 10 broodstock. Tretments included control: polychete worm (P. nunti), Squid nd clf liver (nturl feed); T 1 : pellet feed with 50% crude protein nd 8% crude lipid+ P. nunti, Squid nd clf liver; T 2 : pellet feed with 50% crude protein nd 10% crude lipid+ P. nunti, Squid nd clf liver; T 3 : pellet feed with 40% crude protein nd 10% crude lipid+ P. nunti, Squid nd clf liver; T 4 : pellet feed with 40% crude protein nd 8% crude lipid+ P. nunti, Squid nd clf liver P. nunti; T 5 : pellet feed with 50% crude protein nd 10% crude lipid; T 6 : pellet feed with 50% crude protein nd 8% crude lipid; T 7 : pellet feed with 40% crude protein nd 10% crude lipid; T 8 : pellet feed with 40% crude protein nd 8% crude lipid. Feeding rte (nturl feeds) ws 25% of shrimp broodstock biomss dily (Brock nd Min, 1994). Their composition is detiled (Tles 1 nd 2). In tretments 1, 2, 3 nd 4, 50% of the nturl diets were removed nd pellet foods used insted. The mount of pellet feeds tht given dily, ws 3.5% of shrimp broodstock biomss. In tretments 5, 6, 7 nd 8, the mount of pellet feeds tht given dily ws 7.5% dily. Feeding ws done 4 times dy. Feed types Wet feed Polychete worm (P. nunti), Squid nd clf liver were used for the broodstock feeding nd sexul mturtion. Pellet feed Pellet feeds were mnufctured with lortory equipment. In ddition, depending to tretment type, four pellet feeds were prepred. Anlyses Shrimp length including crpce length nd totl length were mesured by digitl clipers (Model TITAN), wheres, totl wet weight ws mesured using digitl scle (Model WANT) t g nerest (Conides et l., 2008). The sexul, nd weight of ovries, oocyte count, fecundity nd heptopncres weight in femle broodstock were determined (Islm et l., 2012). Also, eyestlk ltion nd sexul development ssessment of broodstock ws performed ccording to Aelgml (2015). Wter temperture, dissolved oxygen, slinity nd ph were mesured dily. Dt were nlyzed using one-wy ANOVA nd the differences mong the tretments were determined using Duncn test (α=0.05). All sttisticl nlyses were performed in SPSS 17.

4 1207 Ghorbni Vghei et l., Production of rtificil diets for femle breeders of western white Tle 1: Proximte composition of the pelleted feeds for the brood shrimps feeding. Food Composition (%) Crude protein Crude ft Ash Fiber Moisture Pellet Pellet Pellet Pellet Tle 2: Proximte composition of the nturl feeds for the brood shrimps feeding. Food Composition (%) Crude protein Crude ft Ash Fiber Moisture Perinereis nunti Squid Clf liver Results Proximte composition of the feed ingredients is shown in Tle 3. The first sexul mturtion ws observed in tretments 2 nd 7. Number of the broodstock in different sexul mturtion s during the experiment is presented in Tle 4. Tle 3: Proximte composition of the feed ingredients. Feed ingredients Composition (%) Crude protein Crude ft Ash Fiber Moisture Shrimp mel Whet mel Soyben mel Fish mel Squid mel Tle 4: Men number of the brood shrimps with different mturtion s during the experiment. Items Control Number Averge 3.05± ± ± ± ± ± ± ±6.66 b 1.24±3.12 c Different letters show significnt difference mong the tretments (p<0.05). In the tretments control, 1, 2, 3 nd 4 were observed, respectively 2, 1, 1, 3 nd 1 brood shrimp in 4 of mturtion. Growth nd reproduction indices re presented in Tle 5 nd Figs The growth indices of ovries in the control, tretment 3 nd tretment 6 were significntly higher thn tht in the other tretments. HSI ws similr in most tretments nd showed some significnt difference. Absolute

5 GSI Irnin Journl of Fisheries Sciences 16(4) fecundity of the control nd tretment 3 ws significntly higher (p<0.05) thn tht in the other tretments. Survivl rte in tretment 8 ws significntly lower (p<0.05) thn tht in the other tretments. Tle 5: Men (±SD) of growth nd reproductive indices in different tretments. Fetures Control Weight )g( 4.77± ± ± ± ± ± ± ± ±37.6 Crpce length 0.13± ± ± ± ± ± ± ± ±2.48 )cm) Body length ) cm) 0.78± ± ± ± ± ±14.96 b 0.77±14.82 b 0.54± ±15.16 In ech row, different letters show significnt difference mong the tretments (p<0.05). Figure 1: Correltion between the tretments nd frequency of vrious s of mturtion b bc cd de e control Tretment f Figure 2: Effect of different tretments on GSI (different letters ove the brs men significnt difference t α=0.05).

6 (%)survivl Fecundity 1209 Ghorbni Vghei et l., Production of rtificil diets for femle breeders of western white Figure 3: Effect of different tretments on HSI (different letters ove the brs men significnt difference t α=0.05) b c cd d b e control Tretment f Figure 4: Effect of different tretments on fecundity (different letters ove the brs men significnt difference t α = 0.05) b control Tretment Figure 5: Effect of different tretments on survivl rte (different letters ove the brs men significnt difference t α = 0.05). There were no significnt differences (p>0.05) in survivl rte nd finl weight of broodstock mong the tretments. The broodstock length in the tretments 5 nd 6 ws significntly lower thn tht in the other tretments (Tle 5).

7 Irnin Journl of Fisheries Sciences 16(4) Men (±SD) of wter physico-chemicl prmeters during the experiment re presented in Tle 6. Tle 6: Men (± SD) of wter physico-chemicl prmeters during the experiment. Prmeters Wter temperture ( ⁰ C) Dissolved oxygen (mg/l -1 ) Slinity (ppt) ph mount 30±1 6.93± ±2 8.16±0.11 Discussion At present, the nutritionl requirements of brood shrimps re hrd to supply in propgtion centers. The min problems with fresh wet feeds (worm nd squid) re their vilility, high price, difficulty of trnsporttion nd wshing prior to use. Thus, in this study 50% nd 100% of nturl wet feeds were replced with pellet feeds to investigte the possibility of the replcement of pelleted feeds to wet feeds. Also, it hs been reported tht totl replcement of nturl feeds with rtificil ones hs not been chieved, yet (FAO, 2010). Severl fctors hve mde it impossible to use lrge mounts of pellet feeds for shrimp breeder mturtion. Chimsung (2014) reported tht polychete worm, improves shrimp reproduction due to hving high nutritionl vlue, rchidonic cid nd importnt reproduction hormones. In ddition, it hs been believed tht polychete worm is n excellent source of not only unsturted ftty cids, but lso sexul hormones similr to those of shrimp. Also, it ws reported tht squids contin high levels of protein, HUFA nd cholesterol, which re necessry for successful growth nd reproduction of shrimp (Chimsung, 2014). In ddition, it hs been reported tht bloodworm, squid, mussel nd clm met long with pelleted feeds should be used for shrimp feeding (Brock nd Min, 1994). Divn et l. (2009) reported tht pelleted feeds contining 50% protein nd 10% lipid (PUFA) cn be used in combintion with fresh feeds. However, the rte of replcement of fresh feeds with pelleted feeds hs not been suggested. Thus, in the present study, the effect of even 100% replcement ws investigted on brood shrimp. Sngprdub et l., (1994) investigted the effect of different feeding regimes on ovry mturtion nd spwning of blck tiger shrimp (Peneus monodon) nd in line with the results of present study, found tht the highest ovry mturtion nd spwning ws relted to the broodstock fed either wet feeds or wet feeds plus pelleted feeds. As, mting occurs predominntly when femle broodstock re in 4 of sexul mturtion, low mting frequency mens few of the broodstock re in 4. Such condition ws observed in the present study considering tht there were 10 broodstock in ech 300-L tnk. In the investigtion of the breeder sexul mturtion, stress delivered by tnk siphoning nd wter replcement should be considered, prticulrly in smll tnks.

8 1211 Ghorbni Vghei et l., Production of rtificil diets for femle breeders of western white In the cse of the importnce of dietry protein in shrimp broodstock reproduction, Wouters et l. (2001) reported tht due to synthesis of new biotic mterils during the sexul mturtion, the nimls need higher protein. Also, they reported tht protein content of rtificil feeds is out 50% depending on shrimp species nd protein source. Wouters et l. (2004) found n increse in ovry protein content during oocyte development nd reproduction. Requirement for dietry lipid depends on HUFA, phospholipids, sterols nd energy supplementtion. For long time it hs been known tht the crustcens hve little ility to synthesis HUFA, phospholipids nd sterols (Sngprdub et l., 1994). Artificil feeds hve seemingly lower EPA (20:5n-3) compred to nturl eeds, thus, they hve lower n-3-hufa nd DHA: EPA rtio. Diets for the shrimp brooders during sexul mturtion should hve high n-3/n-6 rtios. It seems tht, they require dietry phospholipids (Sngprdub et l., 1994). Verstrcte et l. (1995) reported tht successful sexul mturtion of western white shrimp cn be met using freshly-frozen mrine orgnisms without formulted diets. The results of present study showed tht, it is possible to eliminte 50% of nturl diets nd use pellet feed insted. ACE (2003) reported tht for western white shrimp propgtion, wter slinity, temperture nd ph should be 32 ppt, C nd out 8 respectively nd sturted dissolved oxygen of ove 5 ppm is required. Stickney (2000) recommended the following wter properties for sexul mturtion of tropicl shrimp broodstock: slinity=27-36±0.5 ppt, temperture=27-29±0.2 o C, ph=7.8±0.2, nd dissolved oxygen 5±0.5 mg/l -1. Regrding the mentioned cses, in the present study the physic-chemicl fctors of wter were in the right rnge. Acknowledgements This study ws conducted with the support of Irnin Ntionl Science Foundtion. We thnk the foundtion chief, Projects Evluting Director nd other stff. Also, we thnk the Generl Mnger, Reserch Vice-President, Plnning nd Support Vice-President, Aquculture Section Hed, Reserch Coordintion Mnger, Nutrition Group Mnger nd the other stff t the Irnin Fisheries Reserch Orgniztion. Our thnks re lso forwrded to the Generl Mnger, Reserch Vice-President, Plnning nd Support Vice-President of the Irnin Shrimp Reserch Orgniztion nd the other stff t Bndrgh Reserch Sttion. References Aelgml, A., Ovrin development nd eyestlk ltion in white leg shrimp (Litopeneus vnnmei). Fish Consulting Group, pp Alv, V.R., Knzw, A., Teshim, S.I. nd Koshio, S., Effect of

9 Irnin Journl of Fisheries Sciences 16(4) dietry phospholipids nd n-3 highly unsturted ftty cids on ovrin development of Kurum prwn. Nippon Suisn Gkkishi, 59, Automtic Content Extrction (ACE), Tiger prwn (Peneus monodon) nd white legged shrimp (Litopeneus vnnmei). 18P. Brock, J.A. nd Min, K.L., A guide to the common problems nd disese of culture Peneus vnnmei. The Ocenic Institute Mkpuu Point. Thilnd. 241P. Chimsung, N., Mturtion diets for blck tiger shrimp (Peneus monodon) broodstock: A review. Songklnkrin. Journl of Science nd Technology, 36(3), , Conides, A., Glmuzin, B., Dulcic, J., Kpiris, K., Jug-Dujkovic, J. nd Ppconstntinou, C., Study of the reproduction of the Krmote shrimp Peneus (Melicertus) kerthurus in Amvrkikos Gulf, Western Greece. Act Adritic, 49(2), Coutteu, P., Phm, D., Pinon, E nd Blczr, Y., Mixed mturtion diets improve shrimp broodstock performnce. Globl Aquculture Allince, creted P. Divn, A.D., Joseph, S. nd Ayyppn, S., Physiology of reproduction, breeding nd culture of tiger shrimp Peneus monodon (Fricus). Arendr Publishing House, 292P. FAO, The pre-spwning process. Originted by Fisheries nd Aquculture Deprtment. pp Islm, M.R., Sultn, N., Bell Hossin, M. nd Mondl, S., Estimtion of fecundity nd gondosomtic index (GSI) of Gngetic whithing, Sillginopsis pnijus (Hmilton, 1822) from the Meghn River Estury, Bngldesh. World Applied Sciences Journl, 17(10), Mglhes, T., Mossolin, E.C. nd Mnteltto, F.L., Gondosomtic nd heptosomtic indexes of the freshwter shrimp Mcrobrchium olfersii (Decpod, Plemonide) from So Sebstito Islnd, Southestern Brzil. Pn- Americn Journl of Aqutic Sceinces, 7 (1), 1-9. Sngprdub, S., Fst, A.W., Piytirtivorkul, S. nd Mensvet, P., Effects of different feeding regimes on ovrin mturtion nd spwning of pondrered gint tiger prwn in Thilnd. Biotec Publiction Thilnd, pp Stickney, R.R., Encyclopedi of quculture. A Wiley-Interscience Publiction. 1060P. Teikw, E.D. nd Mgy, Y.D., Abundnce nd reproductive biology of the peneid prwns of Bgmoy costl wters, Tnzni. Western Indin Ocen. Journl of Mrine Science, 2(2), Verstrete, P., Mor, B. nd Lvens, P., Mturtion of Peneus

10 1213 Ghorbni Vghei et l., Production of rtificil diets for femle breeders of western white vnnmei by using dry pellets s prtil substitute of the nturl diet Ed. I. (Eds) Lvens, P., Jspers, E., Roelnts. Lrvi 95. Europen Aquculture Society. Wouters, R., Lvens, Nieto, J. nd Sorgeloos, P., Peneid shrimp broodstock nutrition: n updted review on reserch nd development. Aquculture, 202, Wouters, R., Nieto, J. nd Sorgeloos, P., A review of recent reserch on shrimp broodstock nutrition nd rtificil diets. Retrieved from https// www. Feed of broodstock.htm., X.