Seyed-Mehdi Mirheydari, Abbas Matinfar and Hosein Emadi

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1 World Journl of Fish nd Mrine Sciences 6 (4): , 204 ISSN IDOSI Publictions, 204 DOI: /idosi.wjfms Influence of Dietry HUFA on Growth, Survivl, Fecundity, Egg Dimeter nd Ftty Acid Composition of Broodstock White Leg Shrimp (Litopeneus vnnmei) Tissues 2 3 Seyed-Mehdi Mirheydri, Abbs Mtinfr nd Hosein Emdi Deprtment of Fisheries Sciences, Fculty of Agriculture nd Nturl Resources, Science nd Reserch Brnch, Islmic Azd University, Tehrn, Irn 2 Aquculture Center, Irnin Fisheries Reserch Orgniztion, Tehrn, Irn 3 Deprtment of Fisheries, North Tehrn Brnch, Islmic Azd University, Tehrn, Irn Abstrct: This experiment ws conducted to evlute the effect of different levels of dietry Highly Unsturted Ftty Acids (HUFA) in pelleted experimentl diets compring to those in nturl foods s control diet, s well s their potentil interction, on growth, survivl, fecundity nd egg dimeter ccording to the ftty cid composition of heptopncres nd ovry tissues of broodstock Litopeneus vnnmei. All experimentl diets were bsed on similr stuffs, Isonitrogenous (3.5%) nd Isocleric (6.9%) primrily. The HUFA source ws HUFA rich oil clled Espirselco. This experiment tested totl dietry levels of highly Unsturted ftty cid (TDH), 2 nd 3% on Fecundity, egg dimeter, survivl, weight gin (%WG) nd length gin (%LG) compred with nturl diet effects on the sme prmeters mentioned bove. No significnt differences (t P<0.05) mong tretments were observed for survivl 40 dys post-bltion. Shrimps fed diets contining HUFA 3% nd control Diet hd no significnt differences together but significntly higher finl fecundity nd egg dimeter thn those fed diets contining the nd 2% HUFA. Femles fed diets contining 2% HUFA didn t obtin significntly higher fecundity, but higher egg dimeter thn those fed diets contining lower HUFA. The most weight gin (WG) were obtined in those shrimps fed diets contining % HUFA which ws significntly higher thn other tretments but the highest length gin (LG) were obtined in control diet nd diets contining % HUFA, respectively ccording to the quntity. Therefore According to the pssge, Femles fed diets contining % HUFA obtined significntly higher finl growth indices thn other tril tretments. But the most level of HUFA is suggested s dietry HUFA requirements for broodstock Litopeneus vnnmei. Key words: Growth Survivl Shrimp Litopeneus vnnmei Fecundity Egg dimeter HUFA Broodstock INTRODUCTION sfe nd progrmmble method. In this regrd, shrimp is of certin position in this industry s one of the vluble Due to the incresed popultion nd the public trend crustcens. Shrimp culture nd propgtion, which ws to use qutic resources produced foodstuff followed by once quite elementry nd limited to certin regions of the findings their medicl importnce nd role in prevention world is now converted into one of the most importnt nd tretment of mny hrdly-treted diseses, the globl industries []. consumption of qutic is incresing. Such unvoidble The dietry nutritious substnces ply n importnt relity hs cused tht qutic niml husbndry hs role in improvement of the growth nd survivl indices. been considered by most of the countries ll over the Aprt from their quite importnt structurl role, the lipids world in the interntionl responsible orgniztions s ct s the min energy provider fctor in the shrimp s Corresponding Author: Seyed-Mehdi Mirheydri, Deprtment of Fisheries, Fculty of Agriculture nd Nturl Resources, Science nd Reserch Brnch, Islmic Azd University, Tehrn, Irn. Tel: , Mob:

2 World J. Fish & Mrine Sci., 6 (4): , 204 body. The role of ftty cids nd especilly their Highly effective help to improvement the production of such Unsturted type (HUFA) re considerble, either s their species nd continunce of the shrimp cultured importnt role in growth ssocited to the shrimp sexul industry. Achieving such results my lso be n mturity or their vluble effects on the helth of the effective help to blncing the essentil ftty cids in consumers [2]. In previous studies [3, 4] it ws reported shrimp diet to improve the production cpbility of this tht the crustcen s bodies hve limited bility to mke species nd growth nd continunce of shrimp HUFA nd therefore require foodstuff contining such reproduction nd cultured industry nd lso ssist more vluble substnces. helth for the consumers of such vluble nd nutritious The reserch of Knzw & Teshim [3] nd crustcens. Knzw et l. [5] proved tht the Jpnese shrimp (Mrusupeneus jponicus) requires Eicospentenoic MATERIALS AND METHODS Acid (EPA) nd Docoshexenoic Acid (DHA) essentil ftty cids for their growth nd sexul mturity. Experimentl System: A forty dys tril ws conducted Additionlly, n-3 HUFA includes the most essentil ftty from April to June, A totl of 2 conicl tnks cid, which is obtined from Linolenic Acid (LNA) nd 2 (Bottom re 0.55 m, 300 Lit) were used. Ech diet ws Linoleic Acid (LOA), respectively s to the importnce. ssigned in ech tretment with three replictes. A 4-h Tody, the importnce of dietry HUFA for crustcen light nd 0-h drk photoperiod ws provided by mturtion nd reproduction hs been recognized by fluorescent lights controlled by timers. Temperture, Hrrison [6]. Published studies on the quntittive dietry slinity nd ph were controlled t 28.8±.0 C, 32±.0 ppt requirements of Peneid shrimp broodstock re very nd 8.2±0.3 respectively. Dissolved oxygen verged limited, totlly. Most broodstock nutrition in primry 6.58±0.32 mg/l [7]. Ammoni, nitrite, nitrte were studies ws bsed on tril nd error experience or monitored weekly nd verged 0.0±0.08 mg, NH 4 N/l, experimentl reserch using fresh food items. Purified or 0.06±0.04 mg/l NO 2 N/l, 5.87±.9 mg nd NO3-N/l, Semi-purified rtificil broodstock diets which gurntee respectively [8], which controlled by pproximtely 50 % good performnce re yet to be developed, crefully dily wter exchnge. formulted prcticl foods, on the other hnd, my hve performnce close to tht of fresh food [7]. Using the Experimentl Shrimp: The cultured subdult femles enriched pellet diet insted of nturl foods my provide Litopeneus vnnmei were obtined from pound in n effective economicl sving for the shrimp culture Bndrgh Shrimp Institute (Bushehr Province-Irn), prt from the incresed qulity []. whose verge weight ws 3± g [7]. Four subdult In the formulted prcticl diets don t llow precise shrimps were reserved per 300 liter tnk. Treted with control of the levels of certin nutrient [8], however, formlin upon rrivl nd cclimted to the experimentl lipids my be the exception to the rule, the prcticl conditions for pproximtely 2 weeks period. Due to ingredients cn be deftted nd different oils with distinct which they were fed nturl diets (Perinereis worm, ftty cid compositions cn be used for the formultion of Mellis penshell nd cuttlefish) nd commercil feed the experimentl diets [9]. Middleditch et l. [0] nd (Hvoursh-4006) s they fed similr foodstuff, lredy in Lytle et l. [] ttribute the success of fresh food the greenhouse pound. At the end of the dpttion mturtion diets (squid, bivlves, bloodworms) to period ll subdult shrimps were blted [9]. their high HUFA content. As well, dietry n-3hufa hs been shown to improve reproductive performnce in Diet: The bsis of the food ws the Hovoursh-4006 L. vnnmei [2, 3] nd other shrimp species [4, 5]. commercil food, the lst diet which is given to the Alv et l. [6] retrded ovrin mturtion in broodstock shrimp t the end of the culturing step. All M. jponicus fed HUFA-free diet. However, Chu et l. Dried diets were similr in foodstuff, isonitrogenous [2, 5] nd Wouters et l. [3] could not find ny (3.5% protein) nd isocloric (6.9% ft) initilly, But with evidence to support the hypothesis tht high dietry different HUFA levels finlly. To exert better control on HUFA levels promote ovrin mturtion. HUFA levels, broodstock mels (Hvoursh-4006) were The present study imed t determining quntittive deftted with chloroform nd methnol ccording to the HUFA requirements for growth nd mturtion modified version of the method described by Floch et l. performnce (fecundity nd egg dimeter) for cultured [9], then deftted mels where plced in fume hood to L. vnnmei femles. Achieving such results my be n remove residul solvents [9] thus, the HUFA rich oil ws 34

3 World J. Fish & Mrine Sci., 6 (4): , 204 dded to the experimentl diets with respect to, 2, 3% of composition of the test diets ws reflected to certin the diet weight rtios. This oil, which is minly extent in the ftty cid composition of heptopncres combintion of Eicospentenoic Acid (EPA) nd nd muscle tissue of shrimp, the ovries nd Docoshexenoic Acid (DHA), ws Espirselco, products heptopncres took out using the Floch et l., [9] in INVE-Aquculture Co, Belgium. method nd were weighed nd recorded exctly. The Tril shrimps were fed by pproximtely 4% of the heptopncreses nd ovries kept frozen under -40 C, totl shrimp biomss (TSB) ws pplied, t the beginning ftty cids nlysis ws mde using gs chromtogrph of the experiment, while due to their growth, the feeding mchine by Leiboritz et l. [20] method. rte ws incresed to 6% dy, finlly. The experimentl In present reserch method, the ovries were diets were fed to shrimp 4 times dy, every 6 hours nd collected nd froze until they could be weighted nd the feeding rte djusted to provide feed slightly in grvimetric sub smples could be dissected nd weighted. excess. Uneten feed, fecl wste nd molt exuvite were Our trget weight for ech sub smples ws pproximtely dily removed before the first feeding. Also, nturl g. Ech ovry sub-smples were then preserved in food-contining (cuttlefish, mellis bivlve, Perinereis modified Gilson s fluid to promote the brekdown of the worm) were used s the control diet, while their ovrin tissue (mixing proportions; 0.05 nitric cid, consumption percentge with respect to the shrimp glcil cetic cid, % ethyl lcohol, distilled biomss ws 0 % in verge [7]. The effects of the three wter). The sub-smples were kept seprtely in some dry diets should hve been compred with ech other nd smll boxes in drkness nd the smple-contining boxes eventully the control diet. All ingredients were mixed were shken once dy only so tht the seprtion of the with 45% (w/w) heted wter nd the resulting dough ws eggs from the ovrin tissue ws expedited. So the ovry pelleted with met grinder, dried in n oven t 45 C for tissue seprtion my be completed nd the eggs were left 4-h nd kept in seled plstic bgs t -25 C until use [9]. in the Gilson s liquid grdully. After exposure for seven Wter stbility of ech diet ws surveyed bsed on the weeks the seprted eggs of ech sub-smple were Wouters et l. [9] method, were obtined bout 3 hours. counted nd determinte their dimeters in the lbortory In order to determine the type nd quntity of the [23]. ftty cids in the prepred experimented diets, the methyl In order to count the eggs, first of ll three sieves ester method ws used to extrct nd nlyze of the ftty were used, while the 0 micron mesh sieve ws plced cids using the Gs chromtogrph mchine [20]. The between two other sieves with 5 nd 50 micron meshes results hve been given in Tble. [24]. Then ech of the smples were dischrged seprtely on the upper sieve nd then wshed with wter inside Growth nd Survivl: Due to mesuring nd recording the 00ml erlen. In order to count the eggs nd volume-bsed weight nd length of ech shrimp t the beginning nd method ws used [25]. end of the period, the survivl, length gin (LG) nd A Snder microscope equipped with scled weight gin (WG) were the indices used to evlute micrometer ws used to determine the egg dimeter. The shrimp performnce. WG ws clculted from the dimeter of 4-5 eggs were verged nd recorded in ech eqution; % WG=Finl Weight- Initil Weight/ Initil sub- smple [24]. Weight x 00 %, Survivl ws trnsformed by rcsine In order to nlyze the effect of heptopncres nd squre root before sttisticl nlysis. The LG for ech ovry HUFA on fecundity nd egg dimeter, the one-wy dietry tretment ws estimted from % LG=Finl ANOVA ws used nd significnt difference ws Length- Initil Length/ Initil Length 00 %. Dt were obtined between them in the Duncn test in the 5% nlyzed by one-wy fctoril ANOVA using system s possibility level. The nlysis ws mde using the block fctor to determine the effects of HUFA. Duncn s STATISTICA softwre [26]. multiple rnge tests ws used s the men seprtion procedure (P<0.05). Sttisticl nlyses were done using RESULTS the SAS softwre pckge [2, 22]. Ftty Acid Anlyses of Test nd Nturl Diets nd Fecundity nd Egg Dimeter: At the end of 6 week Shrimp Tissues: The results of the nlysis experiment, the treted shrimps took out from the tnks concerning the ARA, EPA nd DHA ftty cids of the nd their lengths were mesured nd recorded. According experimentl nd control diets hve been given in to the results of Gonzlez-Felix et l. [8], the ftty cid Tbles nd

4 World J. Fish & Mrine Sci., 6 (4): , 204 Tble : HUFA composition (% of totl) of rtificil diets of broodstock (L. vnnmei) HUFA ftty cids 3% TDH 2% TDH % TDH C20:4n C20:5n C22:6n Totlly *For clrity only HUFA ftty cids yielding. Tble 2: HUFA composition (% of totl) of fresh foods (nturl diet) of broodstock (L. vnnmei) nd men vlues (±SE) of fresh food mix (cuttlefish/bivlve/worm ::) HUFA Cuttlefish (Sepi officinlis) Bivlve (Mellis sp.) Worm (Perinereis nunti) Men vlues (±SE) C20:4n ± 56.0 C20:5n ± 58.0 C22:6n ± 82.0 Totlly ± 97. *For clrity only HUFA ftty cids yielding. Tble 3: HUFA composition (% of totl) of broodstock (L. vnnmei) heptopncres tissue Artificil diets Fresh food diet 3%TDH 2%TDH %TDH HUFA of Heptopncres % % % % C20:4n ± ± ± ± 0.63 b b b B B B A C20:5n ± ± ± ±.55 b B B C A C22:6n ±.5 3.4± ± ±0.0 B B B A *For clrity only HUFA ftty cids yielding. *Within rw, vlues with common cpitl letters suggest sttisticlly different (P<0.05) while within column vlues, with common superscript letters re not sttisticlly different (P>0.05) The consumption levels of ech of the live foods of cids hve lso been determined using sttisticl methods cuttlefish, mellis bivlve nd Perineris worm duding the (P<0.05) ( Tbles 3 nd 4). period were mesured seprtely, which were 7.5, 0 nd The results of the present study demonstrted tht 3.5 % of the shrimp biomss, respectively. highest dietry HUFA is in the 3%, 2% nd % The ANOVA test showed tht the highest level of tretments, respectively s the quntity (Tble ). heptopncres essentil ftty cids were in HUFA % According to the Tbles nd 2, indicted tht the tretment. However, there ws observed no significnt dietry EPA nd DHA of 3% HUFA tretment were higher difference between other tretments in this regrd (P>0.05) thn the sme for the nturl food diet. (Tble 3). Study on the shrimp tissue s ftty cids indicted The nlysis of the ftty cids in the nturl foods in tht the level of both C22:6n3 nd C20:5n3 in % HUFA Tble 2 indicted tht the level of ll the three ftty cids femle ovries ws significntly lower thn other of the Perinereis nunti worm is higher thn other tretments, but in the nturl food nd 3% HUFA tretments nd the Sepi officinlis cuttlefish ftty cid tretment ovries were significntly higher thn other is lower thn other tretments. tretment ovries (P < 0.05). Also it ws indicted tht the The results of the studies showed the ftty cids level of HUFA of the % HUFA femle s composition of pellets nd nturl food diets which hve heptopncreses is significntly higher thn other been given in Tbles nd 2. Also, for better survey on tretments. the results, prt from giving sttisticl comprison of ll In other hnd the highest bsorption of dietry ovries nd heptopncreses ftty cid types between HUFA in ovries of ll tretment s femles ws C20:5n3 vrious tretments, the highest nd lowest dietry ftty (Tble 5), but C22:6n3 in heptopncreses (Tble 3). 343

5 World J. Fish & Mrine Sci., 6 (4): , 204 Tble 4: HUFA composition (% of totl) of broodstock (L. vnnmei) ovry tissue Artificil diets Fresh food diet 3% TDH 2% TDH % TDH HUFA of ovries % % % % C20:4n6 3.4± /0 ± 9/4 6.4±.2.37± 0.36 c c b B B A C C20:5n3 0.7± /2 ± 56/9 6.88± ±0.90 A A B C C22:6n ±.8 29/ ± 203/6 5.47± ±0.94 b b b A B A B * For clrity only HUFA ftty cids yielding. *Within rw, vlues with common cpitl letters suggest sttisticlly different (P<0.05) while within column vlues, with common superscript letters re not sttisticlly different (P>0.05) Tble 5: Survivl rte, Weight gin, Length gin, Fecundity nd egg dimeter of Litopeneus vnnmei broodstock fed incresing totl dietry HUFA level (TDH): men vlues (±SE) t the end of the study. Men vlues (±SE) Diets Survivl rte %WG %LG Fecundity Egg dimeter Control diet 28.00± b 3.9 ±.26 b ± ± ±. % TDH 00.00± 0.28 c ± ±.48 b ± c 5.07 ± % TDH 00.00±0.28 b 6.3 ±.98 c 26.3 ± 0.80 b 5902 ± b 6.36 ± % TDH 28.00± ±.44 b ± ± ±.76 Within column, vlues with common superscript letters re not sttisticlly different (P>0.05). Growth nd Survivl: No significnt differences mong DISCUSSION tretments were observed for survivl nd no significnt interctions were obtined for ny of the responses. Ftty Acid Composition: Due to the recommendtions However, significnt differences in finl weight gin nd of the Wouters et l. [9]; Middleditch et l. [0] nd length gin were observed t the end of the tril. Shrimps Lytle et l. [] tkes into considertion tht ARA is fed diets contining nturl foods obtined significntly thought to be precursor in the synthesis of higher finl length gin, but lower weight gin thn those prostglndin. Sgi et l. [27] nd Hrrison [28] believed fed pellet diets contining different levels of HUFA, in tht the ARA is pre-mteril from which prostglndin exception 3% HUFA tretment femles whose %WG were is mde, thus in this reserch kind of HUFA enriched oil even lower thn control diet femles. Results lso showed nmely Espirselco ws used in the formultion of tht shrimp fed diets contining % HUFA hd experimented diets where included of n-6 HUFA nd n-3 significntly higher finl weight gin nd length gin, thn HUFA. those fed the experimentl pellet tretments (Tble 5). Growth nd Survivl: Results of the present study Fecundity nd Egg Dimeter: No significnt differences showed no significnt difference between ll tretments, mong control diet nd 3% HUFA tretments were however, shrimps fed nd 2 % HUFA tretments observed for fecundity (P>0.05), while both commonly obtined bit higher survivl thn those femles fed 3% higher significntly thn two other experimentl HUFA nd nturl food tretments (P>0.05). Thus the tretments (P<0.05). survivl ws not only ffected by dietry HUFA but Shrimps fed diets contining 3% HUFA obtined no possibility by other fctors such s individul significntly higher egg dimeter thn control diet, but physiologicl differences. However, in Kontr et l. [29] lower weight gin thn those fed pellet diets contining study on the Jpnese shrimp, it ws known tht the lower levels of HUFA (Tble 5). As well, the HUFA, 2% lowest survivl occurred when the low HUFA Artemi tretments differed significntly from ech other (P < 0.05). ws given to the post-lrv. In Nessens et l. [30] 344

6 World J. Fish & Mrine Sci., 6 (4): , 204 studies mde on the post-lrv of white leg shrimp, the In spit of the equl rtio of heptopncres HUFA diets contining incresing levels of dietry HUFA hd no bsorption in shrimp fed 3% HUFA nd control diet difference s per survivl, while in the control which ws tretments, while shrimps fed control diet were without HUFA ws 0, the survivl significntly decresed obtined significntly higher weight gin thn those (P<0.05). Furthermore, ll wter qulity prmeters were fed 3% HUFA diet (P<0.05), suggests tht there re very stble throughout the current tril nd within the other fctors such s individul physiology recommended rnges for commercil shrimp production differences or some environmentl fctors ffecting [3], which lso my hve contributed to high survivl in on %WG. this feeding tril. Results of the present study showed tht, the lowest Some points seem to be noticeble here: (EPA, 20: 5n3) nd (DHA, 22: 6n3) bsorption, were seen in the heptopncres of the 2% HUFA tretment Results from this study showed tht the shrimps. Thus, due to the proved role of the heptopncres HUFA provide no significnt heptopncres ftty cids in the shrimps growth [8], it interction on survivl of Litpeneus vnnmei my be concluded tht the reson of the lowest length broodstock. gin (%LG) in the 2% HUFA tretment is the filure of the According to the Gonzlez-Felix et l. [8], juvenile L. essentil ftty cids from heptopncres to the body of vnnmei fed 5% dietry HUFA were obtined the shrimp, which negtively ffected the length growth. higher significntly growth nd survivl thn other Such conclusion is confirmed by the study results of tretments fed lower dietry HUFA. However, in the Wouters et l. [9]. On the other hnd, there ws no present study on the L. vnnmei broodstock, significnt difference between the level of bsorbed femles fed % HUFA diet provided significntly HUFA in the heptopncres of the control nd 2 nd 3% higher growth thn other groups; Possibility HUFA tretments, while their %LG index showed series suggests the differences between mturing shrimp to of differences. Such issue my indicted the lck of effect juveniles or even non-reproductive dults. In other of the ftty cids on the length growth nd thus the effect words, the ftty cids trnsferring from the of 2% HUFA tretment is considered s either fortuitous heptopncres to the ovry, which occurs between or resulted from the other unknown elements such s rd th the 3 nd 4 stges of sexul mturtion [33,34], briefly individul physiologicl differences. obtined reltively lower in shrimps fed % dietry Results of the present study showed tht, the HUFA thn other tretments; therefore provided highest bsorption percentge of the EPA, DHA nd higher remined heptopncres HUFA in this ARA ftty cids ws seen in the heptopncres of niml s bodies, s well, hs resulted in higher shrimps fed % HUFA tretment (Tble 3), while the somtic growth thn other tretments. highest WG% ws observed in the sme tretment (Tble According to the fct tht in most of the Peneid 5), which my indicte the direct reltion between these species hve been studied up to now, the best ftty cids nd WG%. According to the studies of HUFA level for the dults growth hs been Knzw nd Teshim [3] on the Mrssopeneus pproximtely % dietry HUFA [4, 32, 35], thus it jponicus shrimp nd Chen nd Tsi [32] on suggests HUFA requirement of Peneid shrimp to Frfntepeneys chinensis shrimp, the best growth level higher growth my be decresed becuse most of in ech of these species were obtined in shrimps fed % TDH shll be used for growth nd of the gond HUFA diets. On the other hnd, no sttisticlly significnt development, while the results of this study interction ws found mong of essentil ftty cids nd confirmed the sme issue. WG% in evluted tretments of the present study due to the following two resons: Fecundity nd Egg Dimeter: Results from this study indicted tht the effects of the diets on the fecundity nd Shrimps fed 3% HUFA nd control diet tretments egg dimeter hve not been similr sttisticlly (Tble 5). were obtined higher HUFA bsorption in Results of the present study observed tht the heptopncres thn those fed 2% HUFA tretment, verge quntity of the EPA nd DHA ftty cids in while the femles fed 2% HUFA tretment were heptopncreses of the nturl food tretment shrimp is obtined significntly higher weight gin thn other lower significntly thn those fed 3% dietry HUFA shrimps fed 3% HUFA nd control tretments (Tble 3). However, shrimps fed the nturl foods (P<0.05). obtined no higher significntly fecundity nd egg 345

7 World J. Fish & Mrine Sci., 6 (4): , 204 dimeter, thn those femles fed 3% HUFA diet (P>0.05) Glncing on the nturl foods nlysis indictes tht (Tble 5). In this regrd, Djunidh et l. [36] who the quntity of ech three types of C20:4n6, C20:5n3, reported incresed sexul mturtion indices occurred in C22:6n3 ftty cids in the Perinereis worm re higher thn the nturl food tretment. This suggests tht the higher other nturl foods (Tble 2). Additionlly, due to the fct contents of other essentil nutritious such s protein ply tht in this reserch the shrimps consumption of the worm significnt role both for the fecundity nd egg dimeter. ws higher thn mellis bivlve nd cuttlefish, this In the experimentl diets of this study, incresing suggest tht if shrimps were merely fed with worm, could levels of dietry HUFA provided, the higher fecundity potentilly obtined higher fecundity nd egg dimeter nd egg dimeter, so tht the shrimps fed 3% HUFA even more thn 3% HUFA tretment. In this regrd, the obtined significntly higher fecundity nd egg dimeter results of some previous studies [38, 4] emphsized on thn other experimentl pellet tretments (Tbles 5) which the more effective role of the worms in comprison with shows the direct correltion between dietry HUFA level other nturl foods in development of the sexul mturity nd ovrin development indices. The results of the Chu indices due to their higher essentil ftty cid levels. nd Quzuguel [37] study on Peneus vnnmei nd The results of this reserch clerly demonstrte tht Alve et l. [6] on the Mrsupeneus jponicus re due to heptopncres nd ovry ftty cids, it suggest lso is very similr to the results of the present study, tht the level HUFA ftty cids in the ovries of % lthough both of mentioned studies the enriched nturl HUFA tretment ws significntly lower thn other food ws used insted of dry enriched food. tretments (Tbles 3 nd 5). However, in the Results from this study provide importnt heptopncres of the sme tretment, the HUFA ftty informtion regrding the utiliztion of experimentl cids were significntly higher thn the other tretments. nd nturl food tretments of Pcific white shrimp Thus, due to the fct tht the lowest fecundity nd egg (L. vnnmei) s fllowing: dimeter ws observed in the % HUFA tretment (Tble 4), potentilly cused by the filure in the ftty During this reserch totl feeding rte of 0% d in cids trnsferring from the heptopncreses to the verge of the TSB ws pplied for control tretment, rd th ovries between the 3 nd 4 sexul mturtion stges. while 5% of TSB for experimentl tretments. In this In conclusion, it cn indicte the importnt role of the study the verge of feeding with nturl food heptopncres lipids in ovry development of tretment ws 0% of TSB (4% d initilly which Litopeneus vnnmei broodstock, which is in greement incresed to 6% finlly) [7], such difference my with the results of the Mourente [34] nd Teshim et l. indicte kind of non-hrmony in the feeding [42], were mde on the Mrsopeneus jponicus nd conditions between the nturl food nd dry food Peneus kerthorus species, respectively. tretments shrimps, while the nturl food In this study, the C22:6n3 ftty cid hd the highest consumption ws relted to their wet weight nd level in the heptopncres of ll the tretments, while in their dry weights re lower nd even close to the the ovries the C20:5n3 ftty cid hs the highest level weights of the concentrted food [38]. (Tbles 3 nd 4). This potentilly indictes the conversion On the other hnd, the totl quntity of three HUFA of the C22:6n3 to the C20: 5n3 upon trnsfer from the ftty cids in the nturl foods is round 2% of the heptopncres to the ovry to develop the ovry diet weight nd in the 3% HUFA diet to 3% of the mturtion indices, while indictes the importnt role of diet weight, i.e. the nturl foods ftty cids contents the C20: 5n3 ftty cid in Litopeneus vnnmei sexul re lower (Tbles nd 2) mturtion. However, in study of Chu nd Quzugeul [37], during the mturition of Fenneropeneus indicus According to the points mentioned bove suggest shrimp, this process ws reversed inversely, C20: 5n3 tht the incresed feeding rte with the nturl diet is trnsferring from heptopncres to the ovry vi possibility compensted by the lower HUFA level in this lengthen of the ftty cid chins nd eventully the diet, cused tht the fecundity nd egg dimeter of these conversion of the sme to the 22C: 6n3, while some of the diets to be close together. In greement Djunidh [39] scholrs such s Knzw et l. [5], considered the nd Chu et l. [40] stted tht the eggs HUFA my not reson of is process s difference in the biochemicl be merely fctor of the size nd number of the eggs in the structure of the shrimp vrious species bodies. nturl food diet, while the other compounds such s In the process of shrimp s sexul mturity in this proteins, vitmins, minerls or nturl food pigments my study s tretments, no significnt effect of the dietry lso ply mjor role in this regrd. nd 2% HUFA tretments were seen on the fecundity nd 346

8 World J. Fish & Mrine Sci., 6 (4): , 204 egg dimeter of the ovry. Thus, it my be sid tht the, higher length nd weight femles hd higher GSI nd 2% HUFA rtios in experimentl diets re not suitble for thus hd higher fecundity potentil. This my be due to the incresed fecundity nd egg dimeter t lest due to incresed food consumption of blted shrimps. three following resons: () Incresing TDH levels resulted in incresing fecundity nd egg dimeter in the CONCLUSION present study. (2) There is proof of the beneficil role of medium to high dietry HUFA levels on the reproduction Due to the ovrin HUFA levels in the different prmeters [9, 2, 4] emphsized on the usefulness of the tretments of this study, TDH level of 3% my be HUFA increse of the diets on the reproduction process recommendble for norml fecundity nd egg dimeter of nd lso the qulity of the next genertion. (3) It is known Litopeneus vnnmei. While, the best TDH level of % tht HUFA cnnot be synthesized novo by Peneid is suggested for (%LG) nd (%WG). Thus, ccording to shrimp [5, 9, 43, 44] or only be synthesized to very the results of somtic nd reproductive indices in the limited extent [45], so the HUFA shll dequtely be white leg shrimp femle broodstock, it is recommended to included in Peneid diets. mke such studies in the future, in the lower TDH level Due to the results concerning the fecundity nd egg thn % for the cultured white leg shrimps nd more thn dimeter, the nturl nd 3% HUFA diets my be suggest 3% HUFA for reproduction of this species broodstock is for the proper ovrin development of the white leg suitble. so in the cse of comprison with the results of shrimp (of course under proper environmentl the present study, possibility the most effective TDH on conditions), which is in greement with results of the somtic nd reproductive indices of white leg Wouters et l. [9] nd Alv et l.[ 6] nd Chu nd (Litopeneus vnnmei) shrimp will obtin. Quzuguel [37]. The biochemicl differences of the body structures of the vrious shrimp species cuses tht even ACKNOWLEDGMENTS type of the fixed nutrition hs vrious effects on the different shrimp s reproduction indices [5]. Thus, the The uthors would like to extend their thnks to results of this study re merely relted to the femle those who hve tken the time to criticlly review this broodstock (Litopeneus vnnmei) nd my not be mnuscript. The uthors wish to thnk Dr. Nser Agh, the generlize to other species. Lbortory of ftty cids chemicl nlysis, Artemi The reltion between somtic growth, fecundity nd Reserch Center, University of Urmi, s well for egg dimeter: In the present study in the white leg shrimp prepring the HUFA enriched oil (Spirselco) nd Mr. femle broodstock Litopeneus vnnmei 40 dys post- Foroutn, Hvoursh fctory, for Prepring the bltion, observed no correltion between the of the experimentl diets nd his useful suggestions. The shrimp s weight gin (WG%) nd their fecundity, s the scientific support of Dneshgostrn-Irnin Co. with the highest WG% ws obtined in % HUFA tretment, while sttisticl clcultions is lso pprecited. the highest fecundity nd egg dimeter ws observed in control nd 3% HUFA, which hd significnt difference to REFERENCES other tretments (P<0.05). The results of the Dutov et l. [46] study my pprove the result of this study, s in tht. Ghesleh-Mrmzi, J., 200. Influences of Unsturted study there ws observed no significnt colition ftty cids on Growth indices of Indin shrimp between the size of the egg nd fecundity while in this (Peneus indicus Milne, Edwrds). Fisheries doctorl study the highest length percentge increse, like the thesis, Fculty of Noor-Mrine sciences, University highest fecundity, ws in the control tretment, which hd of Trbit-Modrres, pp: 62. significnt difference with the dry food tretments 2. New. B., 987. Feed nd feeding of shrimp nd fish, (P<0.05). FAO, Rome, pp: 275. However, Plcios et l. [47] studied the shrimp (WG 3. Knzw, A. nd S. Teshim, 979. Reltionship %) upon cultured conditions nd reported positive between essentil ftty cid requirements of qutic correltion between incresing fecundity nd incresing nimls nd the cpcity for bioconversion of (WG %) dys fter bltion. In greement with linolenic cid to highly unsturted ftty cids. results of, Cebllos-V zquez [48] which proved tht Comprtive Biochemistry nd Physiology, mong the sme ge femle cultured shrimps, those of 63B:

9 World J. Fish & Mrine Sci., 6 (4): , Kym, M., Hirt, M. Knzw, A. Tokiw S. nd 5. Chu, C.L., G. Guzn nd P. Quzuguel, 995. Effect Stio, M Essentil ftty cids in the diet of prwn-iii. Lipid metbolism nd ftty cid composition. Bull. Jpn. Soc. Sci. Fish, 46: Knzw, A., S.I. Teshim nd S. Tokiw, 977. Nutritionl requirements of prwn. Effect of dietry lipids on growth. Bulletin of Jpnese Society of Scientific Fisheries, 43: Hrrison, K.E., 990. The role of nutrition in mturtion, reproduction nd embryonic development of decpod crustcens: review. Journl of Shellfish Reserch, 9: Wouters, R., J. Nieto nd P. Sorgeloos, Artificil diets for peneid shrimp. Globl Aquculture Advocte, 3: Benzie, J.A.H., 997. A review of the effect of genetics nd environment on the mturtion lrvl qulity of the gint tiger prwn Peneus monodon. Aqucuture, 55: Wouters, R., Piguve, X. Bstids, L. Clderon J. nd P. Sorgeloos, 200. Ovrin mturtion nd hemolymphtic vitellogenin concentrtion of Pcific white shrimp Litopeneus vnnmei (Boone) fed incresing levels of totl dietry lipids nd HUFA. Aquculture Reserch, 32: Middleditch, B.S., Missler, S.R. Wrd, D.G. McVey, J.B. Brown nd A.L. Lwrence, 979. Mturtion of Peneid shrimp: dietry ftty cids. Proceeding of the World Mriculture Society, 0: Lytle, J.S., T.F. Lytle nd J.T. Ogle, 990. Polyunsturted ftty cid profiles s Med R.; comprtive tool in ssessing mturtion diets of Peneus vnnmei. Aquculture, 89: Chu, C., J.C. Guilume, G. Stephen nd L. Chim, 994. Influence of phospholipids nd highly unsturted ftty cids on spwning rte nd egg nd tissue composition in Peneus nnmei fed semi purified diets. Aquculture, 26: Wouters, R., L. Gomez, P. Lvens nd J. Clderon, 999. Feeding enriched Artemi biomss to Peneus vnnmei broodstock: Its effect on reproductive performnce nd lrvl qulity, Journl of Shellfish Reserch, 8: Xu, X.L., W.L. Ji, J.D. Cstel nd R.K. Odor, 994. The nutritionl vlue of dietry n-3 nd n-6 ftty cids for the Chinese prwn (Peneus chinensis). Aquculture, 8: of highly unsturted ftty cids, Alph- tocopherol nd scorbic cid in broodstock diet on egg composition nd Development of Peneus indicus. Comprtive Biochemistry nd Physiology, 2: Alv, V.R., A. Knzw, S.I. Teshim nd S. Koshio, 993. Effect of dietry Phospholipids nd n-3 highly unsturted ftty cids on ovrin development of Kurum prwn. Nippon suisn Gkkishi, 59: Broke, J.A. nd K.L. Min, 994. A guide book to the common problems nd diseses of cultured Peneus vnnmei. The Ocenic Institute Publiction, pp: Gonzlez-Felix, M.L., A.L. Lwrence, D.M. Gtlin nd M. Perez-Velzquez, 200. Growth, survivl nd ftty cid composition of juvenile Litopeneus vnnmei fed different oils in the presence nd bsence of phospholipids. Aquculture, 205: Floch, J., M. Lee nd G.H.S. Stnley, 957. A simple method for the isoltion nd purifiction of totl lipids from niml tissues. Journl of Biologicl Chemistry, 266: Leiboritz, B.E., M.L. Hu nd A.L. Tppel, 987. Lipid proxidtion in rt tissue slices: Effect of dietry vitmin E, corn oil-lrd nd menhden oil. Journl of Lipids, 0: SAS Institute, 999. The SAS System for Windows, Softwre Relese 8., Cry, NC, USA. 22. SAS Institute, The SAS System for Windows, Softwre Relese 8., Cry, NC, USA. 23. Friedlnd, K.D., D. Am-Abssi, M. Mnning, L. Clrke, G. Kligys nd R.C. Chmbers, Automted egg counting nd sizing from scnned imges: Rpid smple processing nd lrge volumes for fecundity estimtes. Journl of Se Reserch, 54: Mohmmdi, G.H., 994. Survey on fecundity of white shrimp (Metpeneus offinis), M.Sc thesis of Mrine biology, Fculty of Mrine Sciences nd Technologies, North Tehrn brnch, Islmic Azd University, pp: Holden, M.J., 974. Problems in the rtionl exploittion of elsmobrnchs nd some suggested solutions. In Se fisheries reserch, edited by F.R. Hrden-Jones, London Elek Science, pp: Med, R., R.N. Curnow nd A.M. Hsted, 993. Sttisticl Methods in Agriculture nd Experimentl Biology. London, Chmpen nd Hll, pp:

10 World J. Fish & Mrine Sci., 6 (4): , Sgi, A., J. Silkovsky, M. Fisher-Berkovich nd 38. Glgni, M.L., G. Cuzon, F. Glgni nd J. Gogenheim, A. Donn, 995. Prostglndin E2 in previtellogenic 989. Influence of diet on reproduction of cptivtes ovries of the prwn Mcrobrchium rosenbergii: Peneus indicus. Aquculture, 8: synthesis nd effect on the level of CAMP. Generl 39. Djunidh, S., 989, Fresh nchovies (Stelophorus Comprtive Endocrinology, 00: sp.) s feed dditives for gint prwn (Peneus 28. Hrrison, K.E., 997. Broodstock nutrition in monodon). Interntionl Informtion System for the mturtion diets. Crustcen nutrition, Advnced in Agriculturl Science nd Technology, pp: World Aquculture, 6 (ed. by L.R. Dbrmo, D.E. 40. Chu, C., P. Slen nd M. De Lorgeril, Frmed Conklin nd D.M Akiym), pp: nd wild fish in the prevention of crdiovsculr 29. Kontr, E.K., P. Lvens nd P. Srgeloos, 995. diseses: ssessing possible differences in lipid Dietry effect of DHA/EPA on culture performnce nutritionl vlues. Nutrition, Metbolism & nd ftty cid composition of P. monodon Crdiovsculr Diseses, Elsevier, 4(): postlrve, LARVI95- Fish nd shellfish lrviculture 4. Nscimento, I.A., W.A. Bry, L.J.R. Trujillo nd symposium. Europen Aquculture Society, Specil A. Lwrence, 99. Reproduction of blted nd publiction. Gent, Belgium, 5: unblted Peneus schmitti in cptivity using diets 30. Nessens, E., A. Vn-Huwert, M.L. Cob, consisting of fresh-frozen nturl nd dried S.T. Ownsend, R. Roms, W. Wouters nd P. Lvens, formulted feeds. Aquculture, 99: Dietry n-3 HUFA nd DHA/EPA requirements 42. Teshim, S., A. Knzw, K. Horinouchi nd of (P. vnnmei) postlrve. Lrvi95-fish nd S. Koshio, 988. Lipid metbolism in destlked prwn shellfish symposium, Europen Aquculture Society Peneus jponicus: induced mturtion nd trnsfer (EAS), Specil publiction. Gent, Belgium, 24: of lipid reserves to the ovries. Nippon Suisn 3. Smoch, T.M., A.L. Lwrence nd W.A. Bry, 993. Gkkishi, 54(7): Design nd opertion of n intensive nursery 43. Clrke, A., 982. Lypid synthesis nd reproduction in rcewy system for peneid shrimp. In: J.P. McVey, the polr shrimp Christmus ntrcticus. Mrine (Ed.), CRC Hndbook of Mriculture: Crustcen Ecologicl Progress Series, 9: Aquculture. CRC Press, Boc Rton, FL, 44. Chng, E. nd J. O Connor, 983. Metbolism nd pp: trnsport of crbohydrtes nd lipids. The Biology of 32. Chen, H.Y. nd L. Tsi, 986. Combined effects of Crustcen (ed. by D.E. Bliss), pp: dietry phosphtidylcholine nd cholesterol on the 45. Mourente, G., 996. In vitro metbolism of C-4- growth, survivl nd body lipid composition of polyunsturted ftty cids in midgut glnd nd shrimp Peneus monodon, 96: ovry cells from Peneus kerthurus Forskl t the 33. Teshim, S. nd A. Knzw, 988. Nutritive vlue beginning of sexul mturtion. Comprtive of sterols for the juvenile prwn. Bull. Jp. Soc. Sci. Biochemistry nd Physiology, 5: Fish., 52: Dutov, S.S., L.I. Popov nd A.I. Beglov, Mourente, G., 990. In vitro metbolism of C-4- Fecundity of grss shrimp, Pndlus kesleri polyunsturted ftty cid in midgut glnd nd ovry (Decpod: Pndlide), ner the southern Kuril cells from Peneus kerthurus Forskl t the Islnds. Russin Journl of Mrine Biology, beginning of sexul mturtion. Comprtive l30(3): Biochemicl nd Physiology, 5: Plcios, E., A.M. Ibrr, J.L. Rmorez, G. Portillo nd 35. Knzw, A., S. Teshim nd S. Endo, 978. I.S. Rcott, 998. Biochemicl composition of egg Requirements of prwn, Peneus jponicus for nd nuplii in White Pcific shrimp Peneus essentil ftty Memoirs of the F M., Fculty of vnnmei (Boone), in reltion to the physiologicl Fisheries, Kgoshim University, 28: condition of spwners in commercil htchery. 36. Djunidh, S., M. Wille, E.K. Kontr nd P. Aquculture Reserch, 29: Sorgeloos, Reproductive performnce nd 48. Cebllos-V zquez, B.P., Evlution of offspring qulity in mud crb (Scyll prmmosin) potentil reproductive of Cmeron blnco broodstock fed different diets. Aquculture Litopeneus vnnmei in conditions of lbortory. Interntionl. Kluwer Acdemic Publishers. Printed in Doctorl thesis. Center of Biologicl Investigtions the Netherlnds, : 3-5. of Noroeste. L Pz, B.C.S., Mexico, pp: Chu, A. nd P. Quzuguel, 989, Lipid metbolism of Peneus vnnmei broodstock: Influence of dietry lipids, Europen Aquculture Society, EAS Specil Publiction, 0:

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