Effect of Fishmeal Replacement With Artemia Biomass as a Protein Source in Practical Diets for the Giant Freshwater Prawn Macrobrachium Rosenbergii

Effect of fishmeal replacement with Artemia biomass as a protein source in practical diets for the giant freshwater prawn Macrobrachium rosenbergii NguyenThi Ngoc Anh 1,2 , TranThi Thanh Hien 2 , Wille Mathieu 1 , NguyenVan Hoa 2 & Patrick Sorgeloos 1 1 Laboratory of Aquaculture & Artemia Reference Center, Department of Animal Production, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium 2 College of Aquaculture and Fisheries, CanTho University, CanTho City,Vietnam Corresponden
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  Effect of fishmeal replacement with  Artemia   biomassas a protein source in practical diets for the giantfreshwater prawn  Macrobrachium rosenbergii  NguyenThi NgocAnh 1,2 ,TranThiThanh Hien 2 ,Wille Mathieu 1 , NguyenVan Hoa 2 &Patrick Sorgeloos 1 1 Laboratoryof Aquaculture & Artemia Reference Center, Department of Animal Production, Facultyof BioscienceEngineering, Ghent University, Ghent, Belgium 2 College of Aquaculture and Fisheries, CanThoUniversity, CanTho City,Vietnam Correspondence:  P Sorgeloos, Laboratory of Aquaculture &  Artemia  Reference Center, Department of Animal Production, Faculty of Bioscience Engineering, Ghent University, Rozier 44, B-9000 Ghent, Belgium. Email:  Abstract A30-dayfeedingexperimentwasconductedin160-Lplastic tanks to evaluate the potential use of   Artemia biomass as a protein source in practical diets forpostlarval  Macrobrachium rosenbergii  (initial meanweight of 12.12^12.29mg). Nine isoenergetic andisonitrogenous experimental diets (approximately40% crude protein) were formulated by replacinglevels of the ¢shmeal (FM) protein di¡erence eitherwith dried or frozen  Artemia  (0,25,50,75 and100%).The 0%  Artemia  treatment, in which Peruvian FMwas the only main protein source, was considered tobe the control diet. The results showed that prawnpostlarvae (PLs) fed the FM control diet had a lowersurvival (46%) compared with all  Artemia  diets.Signi¢cant di¡erences ( P  o 0.05) were, however, onlyfound at 75% and 100%  Artemia  protein inclusionlevels (survival of 68^77%). A gradual increase ingrowth performance (live weight gain, speci¢cgrowth rate and total length) of the prawns wasachieved on increasing dietary inclusion of   Artemia protein. Additionally, the size distribution exhibitedthe same response asgrowthperformance. However,prawns fed the frozen  Artemia  diets showed a betterperformance than the ones fed the dried  Artemia diets. It can be suggested that  Artemia  biomass maytotally replace FM in practical diets for PLs of thefreshwater prawn  M. rosenbergii . Keywords:  Artemia  biomass, ¢shmeal, growth,survival, size distribution,  Macrobrachiumrosenbergii Introduction Presently, in Vietnam, the aquaculture sector hasbeendeveloping interms of the culturearea, produc-tion, number of species and degree of managementintensity(Edwards,Tuan&Allan2004).Forexample,the total aquatic production increased almost 7% in2006, while aquacultureproductionincreased14.6%(Huong & Quan 2007). According to the Ministry of Agriculture&RuralDevelopment,Vietnam,theavail-ability of ¢shmeal (FM) is low and the price of im-ported FM from 2007 to the ¢rst 6 months of 2008has increased signi¢cantly (1.1^1.4USDkg  1 ). Mostfeed manufacturers are using expensive importedFM as a protein source for aquafeeds (http://www.¢ Therefore, assess-ment of cheaper or more readilyavailable alternativeprotein sources such as by-products from ¢sheries,processing or other sources that may reduce the useof FM in feeds is necessary.Among the alternative animal protein sources, Artemia  biomass (adult  Artemia ) may be consideredto be a suitable ingredient for replacing FM in ¢shand crustacean diets because of its high nutritionalvalue (Sorgeloos 1980; Le ¤ ger, Bengtson, Simpson &Sorgeloos 1986; Bengtson, Le ¤ ger & Sorgeloos 1991;Lim, Soh, Dhert & Sorgeloos 2001, Maldonado-Montiel & Rodr| ¤ guez-Canche ¤  2005). Frozen adult Artemia  included ina mixed diet improved the repro-ductive performance of   Penaeus vannamei  (Naessens,Lavens, Gomez, Browdy, McGovern-Hopkins, Spen-cer, Kawahigashi & Sorgeloos1997;Wouters, Lavens, Aquaculture Research, 2009,  40,  669^680 doi:10.1111/j.1365-2109.2008.02143.x r  2009 TheAuthors Journal Compilation r 2009 Blackwell Publishing Ltd  669  Nieto & Sorgeloos 2001). In postlarval stages of pe-naeid shrimp and clawed lobster,  Artemia  has beenshown to provide excellent nutrition (Wickins & Lee2002;Tlusty, Fiore& Goldstein 2005). Liveadult  Arte-mia isalsoanidealfoodforornamental¢sh(Lim etal  .2001). According to Naegel and Rodriguez-Astudillo(2004), dried  Artemia  is a well-suited feed for post-larval  Litopenaeusvannamei .Vietnam o¡ers a potential source of   Artemia biomass as aby-product derived from the commercialcyst-oriented  Artemia  pond production systems,whichyieldanaverage Artemia biomass productionof 0.2^0.3tonneswwha  1 (Brands, Quynh, Bosteels &Baert1995; Anh, Quynh, Hoa & Baert1997). However,so far, only small amounts of this by-product havebeen utilized for production purposes. Therefore, theuse of   Artemia  biomass as a protein source in feedsmay contribute to the pro¢tability of   Artemia  farmersand have a positive impact onthe socio-economic as-pects in the coastal area. Thus, the potential of thisproduct should be evaluated further.There has been a very rapid global expansionof freshwater prawn farming since 1995. The totalglobal production of   Macrobrachium  is estimated at750000^1000000tonnesyear  1 by theendof thisdecade; most is produced in Asia, in which China isthe leader, followed by Vietnam and India (New2005). Currently, thegiant freshwater prawn ( Macro-brachium rosenbergii ), which is indigenous to theMekong Delta,Vietnam, is becoming an increasinglyimportant target species for aquaculture. Accordingto Sinh (2008), in 2006, the total culture area of  M. rosenbergii  in the Mekong Delta was 9077ha andproduction increased to 9514tonnes, with111hatch-eries growing postlarvae (PLs) (production:107mil-lionPL) and about 300 millions of PL imported. Macrobrachium  responds very well to extensive andsemi-intensive culture systems, and its culture, espe-cially in rice ¢elds, is considered tohave thepotentialto raise incomes among impoverished farmers andcontribute towards enhancing rural developmentin Vietnam (Hai, Phuong, Hien, Bui, Son & Wilder2003; Phuong, Hai, Hien, Bui, Huong, Son, Morooka,Fukuda&Wilder2006).Becauseoftherecentseriousdisease outbreaks in tiger shrimp ( Penaeus monodon ),prawn culture has been considered to be an alterna-tive to shrimp in low-saline water areas and coastalsaline soils as they can grow comfortably in watershaving a salinity up to 10gL  1 (New 2002; Cheng,Liu, Cheng & Chen 2003). An increase in prawnculture has led to a growing demand for PLs fromhatcheries. Prawn farmers prefer to stock PL olderthan 10 days to ensure a high survival rate in theirgrow-out ponds. However, feed is the single largestcost item for  M. rosenbergii  culture, as it constitutes40^60% of the operational costs (Phuong, Son,Bui, Tuan & Wilder 2003; Mitra, Mukhopadhyay &Chattopadhyay 2005). Moreover, the formulationof well-balanced diets and theiradequate feeding areof utmost importance for successful aquaculture(Watanabe 2002). The nutritive value of formulatedfeeds depends on the ingredient composition andconsiderably a¡ects the prawn performance, espe-ciallyintheindoor-nurseryphase,wheretheprawnsrelysolelyonsupplementedfeed.Hence,developmentof formulated feeds to attain higher survival, bettergrowth and more e⁄cient feed conversion ratiosis needed. The main goal of the study reported herewas to evaluate the use of   Artemia  biomass as ahigh-quality protein source to improve postlarvaldiets for  M.rosenbergii . Materials and methods Experimental diets PeruvianFMwassuppliedbyCATACOCompany,CanTho city; dried and frozen  Artemia  biomass were ob-tained fromtheexperimental  Artemia biomass pondsin the experimental station of Can Tho Universityin Bac Lieu province, Vietnam. Dried  Artemia  mealwas obtained by sun drying thin layers of   Artemia biomass. Other ingredients such as soybean meal,soybean oil, squid oil, gelatine, wheat £our. . . werepurchased from commercial suppliers. The dietaryingredients were analysed for chemical composition(Table1) before the formulationof the diets.Nine experimental diets were formulated by repla-cing 0%,25%,50%,75% and100% of the FM proteinina standarddiet with eitherdried or frozen  Artemia biomass (Table 2). In the 0%  Artemia  treatment, Table 1  Proximate composition (% of dry matter) of theingredients used inthe experimentaldiets IngredientsPeruvianfishmealDried Artemia  mealFrozen Artemia  SoybeanmealWheatflour Dry matter 91.38 86.57 12.44 89.19 88.78Crude protein 59.20 50.76 50.89 47.60 11.62Crude lipid 5.62 9.89 9.93 3.17 1.92Ash 33.29 24.97 23.98 7.38 0.71Crude fibre 0.62 2.50 2.48 6.99 1.25 Artemia biomassas protein source for  Macrobrachiumrosenbergii NTNAnh etal .  Aquaculture Research, 2009,  40,  669^680 r  2009 TheAuthors 670  Journal Compilation r 2009 Blackwell Publishing Ltd,  Aquaculture Research,  40,  669^680  Peruvian FM was the main protein source of mixedingredients. All diets were formulated to be approxi-mately isolipidic, isoenergetic and isonitrogenous(40% dietary protein).The‘ SOLVER ’program in Micro-soft Excel was used to establishthe formulated feeds.Inthis program,theproximate compositionof thein-gredients and those of the diets are preset, in whichthe proportion of FM protein substituted by Artemia protein must be precise. Based onthe compositionof FM and  Artemia  meal, it is therefore possible thatother ingredients (e.g. amount of wheat £our, soy-bean oil and squid oil) vary as well in order to keepthegrosscompositionof theresultingdietsassimilaras possible.The diets were made into sinking pellets(700 m m) using a pellet machine, oven-dried at 60 1 Cand stored at 4 1 C. Chemicalanalysis Proximate analysis (moisture, crude protein, totallipid,¢breandash)oftheingredientsandexperimen-tal diets was carried out according to the standardmethodsofAssociationofO⁄cialAnalyticalChemists(1995). Nitrogen-free extract was estimated on a dryweight basis by subtracting the percentages of crudeprotein, lipids, crude ¢bre andash from100%.The amino acid contents of the test diets wereanalysed by liquid chromatographic analysis (HP,Hewlett-Packard AOS ^ 1090 Merck, Darmstadt, Ger-many) using the conductivity of pre-column OPA( O  -phtal aldehyde) and FMOC (9-£uorenylmethylchloroformate).The fatty acid composition of the experimentaldiets was determined by gas chromatography. Fattyacid methyl esters (FAMEs) were prepared via a pro-cedure modi¢ed from Lepage and Roy (1984). Experimental design A feeding trial was conducted for 30 days in theshrimp hatchery of the College of Aquaculture andFisheries, Can Tho University,Vietnam. The test wasset up as a completely randomized design with threereplicates per treatment.The plastic 160L tanks were ¢lled with 120L of de-chlorinated tapwater. Each tank was providedwithcontinuous aerationand blackplastic nets weredistributed throughout the water column as a sub-strate for PL prawns in order to reduce the opportu-nity for cannibalism. Postlarvae from one singlebatch were purchased from a commercial hatcheryin CanTho city and reared in a1m 3 tank for 5 daysbefore the start of the trial.Atthestartof theexperiment,allPLweredeprivedof food for1day.120PL (mean initialweightof12.16^ 12.29mg)weretransferredintoeachtank.50%ofthetank volume was exchanged every 3 days at 14:00hours. Daily water temperature and pH were mea-sured at 7:00 and 14:00 hours using a thermo-pHmeter(YSI60ModelpHmeter,HANNA instruments,Mauritius). Total ammonia nitrogen (NH 3 /NH 4 1 ), Table 2  Composition (g100g  1 dry matter) of the nine experimental diets Ingredients 0%A 25%DA 25%FA 50%DA 50%FA 75%DA 75%FA 100%DA 100%FA Fishmeal 49.35 37.00 37.03 24.66 24.69 12.33 12.35 0.00 0.00Dried  Artemia   0.00 14.39 0.00 28.77 0.00 43.14 0.00 57.55 0.00Frozen  Artemia   0.00 0.00 14.36 0.00 28.73 0.00 43.11 0.00 57.40Soybean meal 12.34 12.58 12.85 13.36 13.35 13.87 13.86 14.34 14.35Wheat flour 25.08 23.03 22.89 20.98 20.71 18.94 18.52 16.90 16.86Soybean oil 1.68 1.33 1.33 0.97 0.97 0.62 0.62 0.27 0.26Squid oil 1.67 1.32 1.32 0.97 0.97 0.62 0.61 0.26 0.26Lecithin 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00 1.00Vitamin Premix 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00 4.00Gelatin 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50 2.50Cellulose 2.38 2.58 2.73 2.78 3.08 2.98 3.43 3.18 3.37 A,  Artemia ; DA, dried  Artemia ; FA, frozen  Artemia .(1) Vitamin produced by Rhone-Poulenc, namelyVemevit. In1kg of mixture, there are vitamin A:2000000UI; vitamin D 3 :400000UI;vitamin E:12000mg; vitamin K:480mg; vitamin B 1 :800mg; vitamin B 2 :800g; vitamin B 6 :500mg; nicotinic acid:5000mg; calcium:2000mg; vitamin B 12 : 2000mg; folic acid: 160mg; Microvitamin H: 2000:1000mg; vitaminC: 100000mg; Fe 2 1 : 1000ppm; Zn 2 1 :3000ppm; Mn 2 1 : 2000ppm; Cu 2 1 :100ppm; iodine: 20ppm; Co 2 1 :10ppm and some others.(2) Gross energy was calculated based on protein 5 5.56; lipid 5 9.54 and NFE 5 4.20.NFE, nitrogen-free extract. Aquaculture Research, 2009,  40,  669^680  Artemia biomass as protein source for  Macrobrachiumrosenbergii NTNAnh etal . r  2009 TheAuthors Journal Compilation r 2009 Blackwell Publishing Ltd,  AquacultureResearch,  40,  669^680  671  NO 2 -N and NO 3 -N were monitored weekly using aspectrophotometer according to American PublicHealth Association (1998).Prawns were fed four times a day at about 8:00,12:00, 16:00 and 20:00 hours at 15% of their bodyweight.Uneatenfeedandfaecalmatterweresiphonedout before the ¢rst feeding in the morning. Thirtyprawns in each tank were randomly sampled at thestart of the experimentandat10-day intervals duringtheexperimentalperiod.Theamountofdietgivenwasadjusted according to these weight measurements.Prawnswereweighedingroupsof30onadigitalMet-tler scale (Denver Instrument, Denver, Colorado) andthe mean weights were determined. Prawns werethen returned to their srcinal tanks. At the end of the experiment, the survival, mean individual weightandtotal lengthof theprawnwere determined. Performance parameters Performance was evaluated by live weight gain, spe-ci¢c growth rate (SGR), total length and survival,whichwere calculated as follows:Liveweightgain(mg) 5 ¢nalweight  initialweightSGR (%day  1 ) 5 100  [(ln ¢nal weight  lninitial weight)/days of experiment]Survival (%) 5 100  (¢nal prawn number/initialprawn number)The total length (mm) of prawns was measuredfrom the tip of the rostrum to the telsonusing a cali-brated ruler. Statistical analysis Data for all measured parameters were analysedusing SPSSforWindows,Version11.0.Variations fromdietary treatment were compared by one-way ANOVA .TheTukey HSD  post hoc  analysis was used to detectdi¡erences between means. Signi¢cant di¡erenceswere considered at  P  o 0.05. All percentage valueswere normalized through a square root arcsinetransformationbefore statisticaltreatment. Results Waterquality Dailywatertemperaturesrangedfrom27.5to30.5 1 C,and pH £uctuated from 7.2 to 7.8; variations in thelevels of total ammonia nitrogen, NO 2 -N and NO 3 -Nwere 0.02^0.61, 0.02^0.32 and 0.08^0.19mgL  1 ,respectively, throughout the experimental period.Generally, the water quality was not much di¡erentbetween treatments and remained within the suita-ble range for the normal growth of   M. rosenbergii PL (D’Abramo, Ohs, Fondren, Steeby & Posadas 2003;Niu, Lee, Goshima & Nakao 2003). Experimental diets Proximate analysis showed that all diets used in thisstudy were nearly isonitrogenous, isolipidic and iso-caloric (Table 3). However, a slightly lower lipid con-tent in the FM-based control diet compared with the100%  Artemia -based diets was noted.The analysed aminoacid concentrations inthe ex-perimental diets (Table 4) indicated that the dietscontaining  Artemia  protein had, for all amino acids,equal or slightly higher levels than the control diet.In general, these di¡erences among the diets werenegligible.The fattyacid pro¢les of the experimental diets areshown in Table 5. The concentrations of linoleic(18:2n-6), linolenic (18:3n-3), eicosapentaenoic acid,(EPA) (20:5n-3) andarachidonic acid (ARA) (20:4n-6),total monounsaturated fatty acids, polyunsaturatedfatty acids (PUFA), n-3 and n-6 fatty acids in the Table 3  Chemical compositions (% of dry matter) of the nine experimentaldiets Ingredients 0%A 25%DA 25%FA 50%DA 50%FA 75%DA 75%FA 100%DA 100%FA Dry matter 91.91 92.76 90.01 92.07 91.25 90.72 92.61 89.62 92.75Crude protein 39.95 39.87 39.64 39.69 39.79 39.78 39.81 39.91 39.94Crude lipid 7.47 7.93 7.95 8.03 7.96 7.99 8.10 8.78 8.15NFE 29.19 28.40 27.63 26.43 26.96 27.12 27.52 26.74 27.96Ash 20.57 20.69 21.49 21.77 22.04 21.09 21.74 20.73 21.42Crude fibre 2.82 3.11 3.29 4.08 3.25 4.02 2.83 3.84 2.53Energy (kcalg  1 ) 4.19 4.20 4.15 4.11 4.13 4.14 4.17 4.21 4.20 NFE, nitrogen-free extract. Artemia biomassas protein source for  Macrobrachiumrosenbergii NTNAnh etal .  Aquaculture Research, 2009,  40,  669^680 r  2009 TheAuthors 672  Journal Compilation r 2009 Blackwell Publishing Ltd,  Aquaculture Research,  40,  669^680
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