Development of a New Type of Fish Diet, Non-Fish … › bulletin › bull › bull40 › 40-02...Key word: Fish diet, Non-fish meal, Extruded-pellet, Yellowtail, Palatability, Substitute

Development of a New Type of Fish Diet, Non-Fish Meal Extruded-Pellet

NorikoISHIDA＊1,TomohikoKOSHIISHI＊2,TatsuoTSUZAKI＊3,KojiMAENO＊4, SatoshiKATAYAMA＊5,MinoruSATOH＊5,andShuichiSATOH＊6

Abstract:Wedevelopedanon-fishmealdietusingplantand/oranimalproteinmaterials.Threekindsofnon-fishmealdietsandacontroldietcontaining50%fishmealwereprocessed.Inthenon-fishmealdiets,thefishmealwasreplacedwithcommerciallyavailableplantoranimalmaterialsandsupplementedwithtaurineandmaterials formaintainingpalatability.Thesedietswere fedtooneyearoldyellowtail (BW753±96g) innetcages.Therewasnodifference ingrowth,dailyweightgain,dailyfeedingrate,feedconversionratioandproteinefficiencyratioamongthediets.Non-fishmealdietswereprocessed ina factoryanddietarypropertieswerestudiedsuchasuptake,stomachevacuationrate,andcomparativediseaseresistanceinfishfedtheexperimentaldiets. Inaddition,palatabilityof each substituteproteinmaterial for fishwasexaminedandmaterialstoenhancepalatabilityofthenon-fishmealdietswereclarified.Non-fishmealdietshavethepotentialtosupportthegrowthofoneyearoldyellowtail.

Key word:Fishdiet,Non-fishmeal,Extruded-pellet,Yellowtail,Palatability,Substituteproteinmaterial

　InJapan, fishmeal is themainmaterial in fishdietsusedinaquacultureandiscontainedatalevelofover50%.Over90%ofthefishmealusedinfishdietsforaquacultureisimported.Globally,thepriceof fishmealhas increaseddramatically in recentyearsduetoreducedproductioncausedbyfisheryregulation in SouthAmerica and the increasedworldwide demand for aquacultured fish. Theincreasingfishmealpricethreatensof theviabilityof production for Japanese aquaculture farmers.Therefore, anewtypeofdiet that is independentof fishmeal is strongly required.Lowornon-fishmealextruded-pelletshavebeendeveloped. Inourpreliminary study,weprepareddiets containing10%fishmeal,and fed themtoculturedyellowtailtocomparethefishgrowthperformancestoadietcontaining 50% fishmeal. Itwas observed thatfishgrowthwassimilar inbothdietgroups.Threeexperimentalnon-fishmeal extruded-pelletswere

designedand fed tooneyearoldyellowtailwhichwereatanimmatureandrapidlygrowingsize.Wealsostudieddietary,uptake,stomachevacuaterate,palatability, andcomparativediseaseresistanceoffish.Then themeatquality fed theexperimentaldietswasevaluatedusingapanelsensorytest.

Materials and Methods

Experimental diets and processing:Thecontroldietandthreekindsofnon-fishmealdietswerepreparedusingavarietyof ingredients (Table1) (Tsuzakiet al., 2013and2014).Thecontroldiet(FM)contained50%fishmealtoprovideasuitablebalanceofaminoacids.Fishmealwas replacedwith commerciallyavailableplant (PP) andanimal (AP)materials inthenon-fishmeal diets. Soyprotein concentrate,defattedsoybeanmeal,andcornglutenmealwereused for plantmaterials andporkmeal, feather

2015年 1 月30日受理（ReceivedonJanuary30, 2015）＊1 NationalResearchInstituteofFisheriesScience,FRA,Yokohama236-8648,Japan E-mail:larry@affrc.go.jp＊2 MarubeniNisshinFeedCo.,Ltd.,Aichi441-3605,Japan＊3 NationalResearchInstituteofFisheriesandEnvironmentofInlandSea,FRA,Ehime794-2305,Japan＊4 KagoshimaPrefecturalFisheriesTechnologyandDevelopmentCenter,Kagoshima891-0315,Japan＊5 TohokuUniversity,Sendai981-8555,Japan＊6 TokyoUniversityofMarineScienceandTechnology,Tokyo108-8477,Japan

水研センター研報，第40号，39－49，平成27年Bull.Fish.Res.Agen.No. 40，39－49，2015

39

Noriko ISHIDA, Tomohiko KOSHIISHI, Tatsuo TSUZAKI, Koji MAENO, Satoshi KATAYAMA, Minoru SATOH, and Shuichi SATOH

Ingredient FM PP-SP PP-FS AP-SPFish meal 1 50.0 0 0 0Soy bean concentrate 2 0 30.0 30.0 0Defatted soy bean meal 3 3.5 11.0 11.0 8.0Corn gluten meal 4 0 12.0 12.0 9.0Krill meal 0 3.0 3.0 3.0Pork meal 5 0 0 0 30.0Feather meal 5 0 0 0 3.0Blood meal 6 0 0 0 3.0Fish oil 7 15.4 17.0 17.0 14.0Wheat 13.0 10.8 10.8 12.0Defatted rice brain 8 8.5 0 0 1.8Tapioca starch 9 6.6 8.0 8.0 8.0Vitamin mixture 10 2.0 2.0 2.0 2.0Mineral mixture 10 1.0 1.0 1.0 1.0Taurine 0 1.0 1.0 1.0Amino acid mixture 11 0 2.2 2.2 2.2Calcium phosphate 12 0 2.0 2.0 2.0Enzyme mixture 13 (external addition) 0 (0.2) (0.2) (0.2)Skipjack peptide 14 (external addition) 0 (0.5) 0 (0.5)Fish sauce 15 (external addition) 0 0 (0.5) 0

10 See Takagi et al． (2005)．

Table 1． Formulation of the experimental diets (%)

Diet group

1 Fish meal was made from anchovy meal as the main raw material, imported from Peru．2 Soycomil; Archer Daniels Midland Japan Ltd．3 Nisshin Oillio Co., Ltd．4 Japan Corn Starch Co., Ltd．5 Kagoshima Pure Foods Co., Ltd．6 APC Japan Co., Ltd．7 Tsuji Oil Mills Co., Ltd．8 Suzuki Oil Mills Co., Ltd．9 Sanguan Wongse Industries Co., Ltd．

11 See Aoki et al．(2000b)．12 Onoda Chemical Industry Co., Ltd．13 Alltech．14 Marubeni Nisshin Feed Co., Ltd．15 Sakaiovex Co., Ltd．

Table 1.Formulationoftheexperimentaldiets(%)

40

Non-Fish Meal Extruded-Pellet

meal,andbloodmealwereusedforanimalmaterialsin this study.Taurinewas added to thenon-fishmealdietsata1%composition.Experimentaldietswerecarefully shapedusinganextruder tohavesuitablephysicalpropertiesintheseaandsinkatanappropriaterateratherthanfloat.　It i s known that ye l lowta i l become lessresponsive to a diet as the amount of fishmealincluded is reduced (Aokiet al., 2000aand2000b).Therefore, two commercially availablematerials,askipjackpeptide (SP)anda fishsauce (FS)weresupplementedtomaintainthepalatabilityofthenon-fishmealdiet.Theskipjackpeptideisconcentratedfromthecookingwaterofskipjack.Thefishsauceismade frommackerelgutusingapatent-pendingmethod for rapidproduction.Anenzymesolutionandaminoacidchelatedtracemetalswereaddedtothenon-fishmealdiets to improvedigestibilityandabsorption.Thesepalatabilitymaterialsandenzymesolutionwereaddedbya spraymethodafter theextruderprocessingtominimizethevolume(Fig. 1).　Therefore, the threekindsofnon-fishmealdietswereasfollows:　　PP-SPwas a plant protein based dietwith

theskipjackpeptideasamaterial toenhancepalatability.

　　PP-FSwasaplantproteinbaseddietwiththefishsauceasamaterialtoenhancepalatability.

　　AP-SPwasananimalproteinbaseddietwiththeskipjackpeptideasamaterial toenhancepalatability.

　Twosizesofexperimentaldiets, 13mmand15mmgraindiameter,werepreparedat theKagoshimaplant ofMarubeniNisshinFeedCo., Ltd.Therewereminimaldifferencesinthegeneralcomposition

amongthefourexperimentaldiets.Smalldifferenceswereseeninthefattyacidcomposition.AP-SPhad16%ofn-3PUFAwhiletheotherdietsrangedfrom22-27%.AP-SP also had 41% of totalmonoenes,higher than the 30-35% found in the otherdiets(Table2). Feeding experiment:Oneyearoldyellowtailswhichoriginated fromseedlingproduction inGotoIsland,NagasakiPrefecturewereused in theexperiments(Tsuzaki et al., 2013and2014).Theaveragebodyweight of 220 fishwas 753±96g.Fourgroups of55fish feddifferentexperimentaldiets tosatiationthree times aweek for sixmonths from July toJanuarywerereared in four4mx4mreplicatenet cages on the sea surface.After January fishcontinued tobe fedwith the leftoverexperimentdietuntilMarch.　The feeding experimentwasdivided into twoperiods, summer season (July toSeptember) andautumnseason(OctobertoDecember).Samplingandanalysiswereperformedforeachperiod.　Body weight and fork length were monthlymeasuredfor20-25 livefishatrandomfromeverygroup. At theendof theexperiment,all the fishweresampledandweighedandthe followingdataweremeasuredorcalculated.　　Conditionfactor＝(W/FL3)×103

　　Weightgainratio (%) ＝ (TWt ＋ SDW－TW0)/TW0×100

　　Dailyweightgainratio(%/day)＝(TWt＋SDW－TW0)/[(TWt＋SDW＋TW0)/2×t]×100

　　Daily feedingrate (%/day)＝F/[(TWt＋SDW＋TW0)/2×t]×100

　　Feedconversionratio(FCR)＝F/(TWt＋SDW－TW0)

　　Proteinefficiencyratio (PER)＝ (TWt＋SDW－TW0)/P

　　W:Bodyweight(g)　　FL:Forklength(cm)　　TW0:Initialtotalbodyweight(g)　　TWt:Finaltotalbodyweight(g)　　SDW:Totalbodyweight of sampled fishand

deadfish(g)　　Nt:Numberoffinalfish　　F:Totalfeedintake(g)　　t:Rearingdays

Fig. 1.Method to addpalatabilitymaterials andenzymesolutiontofishpellets.Rightissprayedafterbeingshapedtoreducethematerialvolume.Leftisshapedafterblendingallthematerials.

Cross-section Cross-section

SprayBlend with materials

41

Noriko ISHIDA, Tomohiko KOSHIISHI, Tatsuo TSUZAKI, Koji MAENO, Satoshi KATAYAMA, Minoru SATOH, and Shuichi SATOH

Analysis FM PP-SP PP-FS AP-SP

Moisture 7.6 7.7 8.8 4.9Crude ash 9.1 5.6 5.6 7.4Crude protein 39.8 38.1 37.4 42.0Crude lipid 18.9 16.1 17.4 17.6

Phosphorus contentP （mg/g） 15.4 9.7 9.7 13.4

Fatty acid composition (%)14: 0*1 4.8 4.2 4.3 5.416: 0 18.3 16.5 18.3 15.818: 0 3.8 3.5 4.1 4.2

16: 1n-7 5.8 5.2 5.5 5.518: 1n-9 16.0 16.8 19.0 16.420: 1n-9 2.6 2.7 4.5 7.520: 1n-11 2.2 2.3 1.5 1.522: 1n-11 4.0 4.1 4.5 9.8

18: 2n-6 4.5 7.0 7.1 5.620: 4n-6 1.0 0.9 0.9 0.4

18: 3n-3*2 1.0 1.2 1.1 0.918：4n-3 1.8 1.5 1.3 2.020: 4n-3 0.6 0.6 0.6 0.620: 5n-3 (EPA) 8.0 6.8 6.0 5.222: 5n-3 (DPA) 2.0 1.9 1.9 1.022: 6n-3 (DHA) 13.5 13.6 11.3 6.2Others 10.2 11.1 8.0 12.2n-3 HUFA 25.9 24.3 21.2 14.9

*2 “n-3”represents the omega-3 fatty acid.

n-3 HUFA＝18: 4n-3+20: 4n-3+EPA+DPA+DHA.Experimental diets are shown in Table 1.

Table2. Proximate composition，phosphorus content, and fatty acid composition ofthe experimental diets

*1 Left figure represents the number of carbonatoms and right figure after colonrepresents the number of carbon unsaturated bonds.

Proximate composition (%)

Diet group

Table 2. Proximate composition, phosphorus content, and fatty acid composition of theexperimentaldiets

42

Non-Fish Meal Extruded-Pellet

　　P:Totalproteinintake(g)　Blood,muscle, and liver sampleswerecollectedfrom5fish ineverygroupatthestart (initial), latesummer (intermediate),and lateautumn (end).Theratioofliverweightperbodyweightwascalculated.Hematocritvalueandplasmacomponentlevelswereanalyzed.Generalcontent,phosphoruscontent,fattyacid composition, and amino acid composition ofmusclewereanalyzed.　ThestatisticaldifferencesbetweentrialdietsweredeterminedbyTukey’smultiplerangecomparisontest(p<0.05). Experimental diet uptake, stomach evacuate rate, and comparative disease resistance of fish fed experimental diets : Nutritional absorption wasanalyzedfromfishfecescollectedfromfishrearedinaconicaltank(Fig. 2)usingspecialdietscontainingchromeoxideasaninternalstandardmaterial(Satohet al., 2013).　Fishwereanesthetizedandadministeredthediet

at 2%of bodyweight directly into the stomach.Stomach residuewas analyzed over timeby theKjeldahlnitrogenmethodandstomachevacuationratewasestimated(Tohataet al., 2012).　Disease resistance was carried out usingStreptococcus dysgalactiae,aninfectiousstreptococcalbacterium.Fishwererearedoneachdiet in indoortanksfor3monthsandfedthebacterium-containingfeedfor5days.Fishmortalitywasobservedovera40dayperiod(Ishidaet al., 2013). Diet palatability:Fishbehaviorwasdividedintofiveresponses afterplacing thediets into a fish tankdescribedasrecognition,ingestion,orientationwithinthemouth,swallowing,orspittingout (Fig. 3).Theactionswereanalyzedbyusingacameratofilmredseabream,yellowtail,andcohosalmonusingsingledrypelletsmade fromonly onematerial (Fig. 4) (Niinumaet al., 2012,Yamauchiet al., 2012).Meat quality of raw sashimi:Apaneltestusingtheordermethod,color-differencemeteranalysis, andelectronicnosesystemanalysiswereperformedforeachexperimentalfishfedPP-SP,PP-FS,AP-SP,andcontroldiet(Tohataet al., 2013).

RESULTS

Processing:Alltheexperimentaldietswereprocessedusingadouble-screwextruder.Atthebeginning intheautumnseason,plantproteinbaseddietsdidnotsinkatasuitablerateandflowedout fromthenetcagecompared to theotherdiets.Therefore thesedietswerere-made.Palatability ingredientsandanenzymesolutionweresprayedontodietstominimizematerialsusedto0.05%fromtheoriginal2%requiredintheblendingmethod(Fig. 1).Growth performances and biological indices in yellowtail:Thegrowthperformances inboth thesummerandautumnperiodareshowninTable3.　In the summer per i od when the wate rtemperaturewas 22.8 - 29.2 ℃ all diet groups

Fig. 2.Theconicaltankusedtocollectfecesoffishfed the experimentaldiets to analyzenutritionalabsorption.Right:Fecesatthebottomofthetank

Fig. 3.Fishbehaviorsforthediet.

Waiting Recognition Orientationwithin the mouth

Ingestion Swallowing

Spitting out

43

Noriko ISHIDA, Tomohiko KOSHIISHI, Tatsuo TSUZAKI, Koji MAENO, Satoshi KATAYAMA, Minoru SATOH, and Shuichi SATOH

showedasimilartotalfeedintakerangingfrom113to118kgandasimilarbodyweightgain (Fig. 5).Intheautumnperiodwhenthewatertemperaturewas11.5 ℃lowerthanduringthesummerperiodtotal food intakeofallgroupswas less than in thesummer ranging from90 to 103kg.No seasonaldifference inweight gainwas seen among dietgroups.

Fig. 4.Behavioralanalysisoffishafterfeedingthediets.

Diet

Filming

Red sea bream, yellowtail, coho salmon

Table 3.Growthperformanceandfeedutilizationbyyellowtailfedtheexperimentaldiets

　Final fork lengthandbodyweightwere48.4cmand2,242g in thecontrol (FM), 48.6cmand2,178g in thePP-SP, 49.0cmand2,234g in thePP-SF,and48.5cmand2,205g in theAP-SP.Differencesbetweencontrol andnon-fishmealdietswerenotobserved and the growthwas almost the same.Therewasno seasonal difference betweendailyweightgain(%/day),dailyfeedingrate(%/day),and

Rearingseason(days)Initial

Summer(90days) Autumn(91days)FM PP-SP PP-FS AP-SP FM PP-SP PP-FS AP-SP

Numberoffish 220＊1

Dead 0 0 1 2 1 0 1 1Sampled 3 4 3 4 7 7 7 7Final 52 51 51 49 44 44 43 41

Totalfoodintake(kg) － 113.0 113.9 113.6 118.0 98.5 102.9 101.0 90.5

Bodyweight(g)＊2 753±96Final 1663±204a 1700±221a 1698±156a 1688±204a 2242±333a 2178±285a 2234±244a 2205±222a

Forklength(cm)＊2 36.9±1.5Final 44.8±2.3a 44.8±1.8a 45.0±1.2a 44.1±1.2a 48.4±2.2a 48.6±1.8a 49.0±1.7a 48.5±1.5a

Conditionfactor＊2 15.0±0.8Final 18.3±1.1b 18.8±0.9b 18.6±0.7b 19.7±1.4a 19.6±1.2a 18.9±1.2b 19.0±1.1ab 19.3±1.0ab

Weightgainratio(%)＊3 － 116.4 119.9 119.1 113.6 34.2 30.4 31.0 32.3Dailyweightgain(%/day)＊3 － 0.82 0.83 0.83 0.80 0.32 0.29 0.30 0.31Dailyfeedingrate(%/day)＊3 － 1.92 1.91 2.02 1.91 1.08 1.15 1.05 　 1.12Feedconversionratio＊3 － 2.35 2.29 2.30 2.51 3.33 3.91 3.76 3.39Proteinefficiencyratio＊3 － 1.07 1.14 1.16 0.94 0.75 0.67 0.71 0.70＊1Fishesweredividedinto4dietgroups,eachgroupcontains55fishes.＊2Dataareshownasmean±standarddeviation(n=20-44)．Valueshavingdifferentsuperscriptlettersaresignificantlydifferent(P＜0.05).＊3Showninthetext.ExperimentaldietsareshowninTable1.

44

Non-Fish Meal Extruded-Pellet

weightgainratio (%).Forexample, feedconversionratio in theautumnseasonwas3.33 in thecontrol(FM)and3.91 inPP-SP, 3.76 inPP-FS,and3.39 inAP-SP.　Table4showstheresultsofthebiologicalindicesofculturedfish(n=5).Thelivertobodyweightratioin everygroupwas 1.2 to 1.3 andno significantdifferencewasseen.Theaveragehematocritvalueof 5 fish ineverygroupwas46.2 to50.6withnosignificantdifferenceobserved.Therewere three,two,andonemortalitiesoffishinAP-SP,PP-FS,andFMrespectively.Greenliver,whichisthoughttobeasignoftaurinedeficiency(Takagiet al., 2005),was

Fig. 5. Growth curves of yellowtail fed theexperimentalfishmealdiet(FM： ● )andnon-fishmealdiet (PP-SP： ○ ,PP-FS： ○ ,andAP–SP：△ )innetcagesfor6months．Experimentaldiets

aredetailedinTable1.

0

500

1,000

1,500

2,000

2,500

Aver

age

body

wei

ght (

g)

Aug Oct Dec Jan

Aver

age

body

wei

ght (

g)

MonthJul Sep Nov

Table 4.Changes inhepatosomatic index,heamatocritvalue,andhematochemicalcharacteristics inyellowtail(n=5)fedtheexperimentaldeits

notseeninanyofthedeadfish. 　Plasmacomponentsmeasured includedglucose,totalprotein, total cholesterol,glutamatepyruvatetransaminase(GPL),andalkalinephosphatase(ALP).No significantdifferencewasobserved inplasmacomponentsbetweenthetreatmentsinthesummerperiod.Total cholesterol exhibited a significantseasonalvariabilitybetween the treatmentswithlowervaluesof246and266mg/dlinPP-SPandPP-FSrespectively(p<0.05) (Table4).Characteristics such as uptake, stomach evacuate rate, and disease resistance of fish fed the experimental diets:Theuptakeofproteinfromnon-fishmealdietswasreducedby4-9 %comparedtotheFMcontrol(Satohet al., 2013).　Evacuation of feed from fish stomachs wasobservedforallthreediets.TheorderoftherateofevacuationwasPP-SP>AP-SP≈FM(Fig. 6).　Survivalrateat42daysafter thechallengetestwas80%inFMgroup,and95%inPP-SPandAP-SPgroup(Ishidaet al., 2013).Diet palatability:Fishspittingout feedwasalmostzero for singledry fishmealpellet.The spittingratewas higher in single dry pelletmade fromplantproteinthanthosemadefromanimalprotein.To improvepalatabilityofPPandAPpelletswewashed theoriginalmaterialswith80%ethanol toremove theextractivecomponentandmadeeachsingledrypellet fromthosepre-treatedmaterials.Thespittingrateinsingledrypelletmadefrompre-

Rearingseason(days)Initial

Summer(90days) Autumn(91days)FM PP-SP PP-FS AP-SP FM PP-SP PP-FS AP-SP

Hepatosomaticindex(%)＊1 1.1±0.0 1.4±0.1a 1.2±0.1a 1.2±0.2a 1.3±0.3a 1.4±0.2a 1.3±0.1a 1.2±0.0a 1.2±0.1a

Hematocritvalue(%) 53.6±3.4 61.5±7.4a 50.8±0.7a 57.0±4.9a 55.3±7.3a 46.2±2.1a 45.7±4.5a 50.6±6.5a 48.0±6.2a

Heamatochemicalcharacteristics　Glucose（mg/dl） 　144±25.2 237.8±51.8a 262.6±30.8a 267.0±41.3a 280.8±146.1a 138.6±10.5a 127.6±21.3a 143.0±12.1a 130.0±28.5a

　Totalprotein(g/dl） 4.0±0.5 4.3±0.7a 3.7±0.2a 4.3±1.5a 4.8±0.7a 　4.1±0.5ab 3.7±0.2b 　3.9±0.5ab 4.4±0.4a

　Totalcholesterol（mg/dl) 349.4±63.0 282.0±30.8a 203.4±21.7a 267.0±79.5a 276.4±101.0a 396.8±72.3a 245.6±29.0b 266.4±61.0b 346.0±82.1ab

　GPT(IU/l)＊2 25.4±2.6 32.4±4.2a 59.2±44.1a 34.6±14.5a 44.4±19.9a 32.0±13.4a 30.8±14.8a 23.4±8.6a 33.8±11.4a

　ALP(IU/l)＊3 148.6±33.8 156.4±34.3a 134.8±14.4a 181.0±64.0a 200.6±22.9a 143.4±43.4a 125.2±16.9a 126.5±23.7a 147.8±42.3a

Mean±standarddeviation(n=5)．Valueshavingdifferentsuperscriptlettersaresignificantlydifferent(P＜0.05）.＊1Theratioofliverweighttobodyweight.＊2Glutamate-pyruvatetransaminase.＊3Alkalinephosphates.ExperimentaldietsareshowninTable1.

45

Noriko ISHIDA, Tomohiko KOSHIISHI, Tatsuo TSUZAKI, Koji MAENO, Satoshi KATAYAMA, Minoru SATOH, and Shuichi SATOH

treatedplantproteindecreased.Ontheotherhand,theoppositeresultwasshowninpre-treatedanimalproteinsthatthespittingrateincreased(Niinumaet al., 2012,Yamauchiet al., 2012). Fish meat analysis:Table5showstheresultsofthefishmeatanalysisofculturedfish.Intheproximatecompositionofall thedietgroups,moisturecontentdeclined and fat content increaseduntil the endof experiment. No difference was seen in thephosphorus content among thediet groups.TheEPA,DHAandn3HUFAcontentoffishfedAP-SPpelletswasalways lowerthan infish fedtheotherdiets, at times significantly less (p<0.05) in bothsummerandautumn.Theseresultsweresimilartotheanalysisofexperimentaldiets.

0

20

40

60

80

100

0 5 10 15 20

FM

PP-SP

AP-SP

（%）

（h）

Fig. 6.Evacuationofthedietfromthefishstomachafterasingleoraladministrationofdietat2%bodyweight.Valueareshownthepercentageofnitrogenin thestomachresidueperadministerednitrogen.ExperimentaldietsareshowninTable1.

Discussion

　Ithas longbeen thought that fishmealwas anecessary ingredient in fish feed.Generally, theformulated feeds foryellowtail containabout50%fishmeal. Recently, formulated feeds containinglower amounts of fish meal such as 30% havebeenproducedasacommercialproduct.Howevera fishmeal content of less than30%wouldhaveconsiderable financial benefits for aquacultureenablingcontinuedeconomicallyviableproduction.Therefore,wedevelopednon-fishmeal feedsandevaluatedthoseinafieldexperimentwithasimilaraquaculture environment.Weused oneyear oldyellowtailwhich is an intermediate size duringculturingandshowsrapidgrowthrequiringhighfeedcosts.Experimentalperiodsincludedthehigherfeedingactivity seasonof summerand the lowerfeedingactivityseasonofwinter. 　In thepresent study (Tsuzaki et al., 2014), theplantproteinbaseddietsofPP-SPandPP-SPshowedasimilardaily feedingrateanddailyweightgain,better feedconversionratioandproteinefficiencyratio than the controlFM in summer season.Onthe otherhand, the animalproteinbaseddiet ofAP-SP showed a higher daily feeding rate andfeedconversionratio, lowerproteinefficiencyratiodemonstrating lowerproteinuptake thanothers.Wealsostudieduptakeofnutrientinthesenon-fishmealdietsandFMbyanalysisofyellowtailfeces.Itisclearthattheuptakeofprotein inthesenon-fishmealdietswerereducedby4- 9 %comparedtothecontrolFM.　Then, in the autumn season, theplantproteinbased diets of PP-SP and PP-FS showed a lowperformancebecauseofthehigherdailyfeedingrateand feedconversionratio, lowerdailyweightgain.Theseresultsarethoughttobeduetothephysicalpropertiesof thediets.Becausetheseplantproteinbaseddietsdidnot sinkappropriatelyand flowedout fromthenetcagecomparedtotheotherdiets.Fromtheseresults,morestudiesareneededfortheimprovement of uptake for animal proteinbaseddietsandadjustmentofthephysicalcharacteristicsfor plant protein based diets. However, theseresultsdemonstrate thatnon-fishmealdietshavethepotentialtosupportthegrowthofoneyearold

46

Non-Fish Meal Extruded-Pellet

yellowtailcompared to theresultsofTakagiet al. (2013). 　For substitutematerials, plantproteinmaterialsuch as soy bean concentrate has been studied(Aoki et al., 2000a, 2000b).Wealsousedsoybeanconcentrate and confirmed its performance as asubstituteproteinforfishmealinthepresentstudy.In the field, it is sometimes believed that plantproteinmaterials stay longer in thestomach thanfishmeal.However, in our experiment, theplantproteinbaseddietsdidnotstay longeramongthedietgroups.Fishmealbaseddietwas the first tobeevacuated from the stomach. Inaddition, it issometimebelievedthatplantprotein fedyellowtailismore susceptible to epidemicdisease than fishmealbaseddiet.However, inourexperiment,thosephenomenawerenotseen (Ishidaet al., 2013).Wealsocomparedthestomachevacuationrateandthe

in vitrodigestionusingonlyoneproteinmaterialdiet, and foundminimaldifference (Tohata et al., 2012).Morestudiesareneeded in futureusing lowcostmaterialssuchasdefattedsoybeanmeal,cornglutenmealandcanolaoil.　As forsubstituteanimalproteinmaterials, therearemeatmeal,chickenmeal,feathermeal,andporkmeal.Feathermealandmeatmealwerereportedtobeusefulforyellowtail(Shimenoet al., 1993a, 1993b, 2000). However, the animal protein studies arefewerthanplantproteinstudies.There ispotentialtousechickenmealandporkmeal.As therearefewstudiesaboutporkmeal,wetried touseporkmealinthepresentstudy.PorkmealhaslessDHA,DPAandn3HUFAoffattyacidthanFM.Asimilartendencywasseen in thewholefishbodyandthemeat of yellowtail fed byporkmeal baseddiet.However,thereisnosignificanteffectongrowthand

Initial FM PP-SP PP-FS AP-SP FM PP-SP PP-FS AP-SPProximate composition (%)

Moisture 71.9 ± 0.8 64.7 ± 1.7 64.5 ± 1.4 64.8 ± 1.7 65.6 ± 4.1 60.6 ± 4.6 60.0 ± 3.0 60.3 ± 1.3 62.5 ± 0.9Crude ash 2.3 ± 0.5 1.3 ± 0.3 1.3 ± 0.0 1.4 ± 0.1 1.4 ± 0.1 1.4 ± 0.1 1.3 ± 0.1 1.4 ± 0.2 1.4 ± 0.1Crude protein 22.1 ± 0.2 21.4 ± 0.6 20.3 ± 1.1 20.5 ± 0.8 20.7 ± 1.3 20.1 ± 2.3 20.5 ± 1.6 22.0 ± 1.0 22.0 ± 0.5Crude lipid 3.7 ± 0.9 11.5 ± 2.1 12.8 ± 2.6 10.6 ± 1.2 10.1 ± 3.3 16.0 ± 6.2 17.9 ± 4.7 16.0 ± 1.2 14.3 ± 1.5

P (mg/g) 15.5 ± 4.4 4.2 ± 1.7 5.5 ± 2.8 5.0 ± 1.5 4.3 ± 2.5 4.6 ± 1.1 4.5 ± 1.6 4.4 ± 0.7 4.7 ± 1.4

Fatty acid composition (%)14: 0 4.2 ± 0.4 4.0 ± 0.3 3.1 ± 0.6 3.4 ± 0.7 3.9 ± 0.2 4.3 ± 0.2 3.9 ± 0.4 3.8 ± 0.3 4.2 ± 0.216: 0 19.6 ± 0.9 18.0 ±0.8 16.2 ± 1.3 16.8 ± 1.1 16.1 ± 0.3 16.4 ± 0.4 16.5 ± 0.7 15.6 ± 0.6 16.0 ± 0.518: 0 5.6 ± 0.5 4.2 ± 0.3 4.0 ± 0.1 4.2 ± 0.5 4.5 ± 0.4 3.4 ± 0.1 3.4 ± 0.1 3.3 ± 0.1 3.9 ± 0.0

16: 1n-7 5.9 ± 0.8 6.6 ± 0.3 5.8 ± 0.5 5.9 ± 0.4 5.9 ± 0.3 7.0 ± 0.3 6.3 ± 0.2 6.2 ± 0.2 6.6 ± 0.318: 1n-9 21.4 ± 1.4 23.2 ± 2.2 25.0 ± 0.8 25.6 ± 1.3 25.0 ± 1.9 20.5 ± 1.9 23.5 ± 1.6 22.6 ± 2.0 22.1 ± 0.320: 1n-9 3.1 ± 0.8 5.1 ± 0.3 5.4 ± 0.3 5.7 ± 0.5 9.4 ± 1.4 5.6 ± 1.2 5.3 ± 0.1 5.6 ± 0.3 6.5 ± 0.220: 1n-11 0.6 ± 0.9 0.2 ± 0.0 0.2 ± 0.0 0.2 ± 0.0 0.3 ± 0.0 0.6 ± 0.9 0.2 ± 0.0 0.2 ± 0.0 2.9 ± 0.122: 1n-11 2.7 ± 0.3 3.4 ± 0.4 3.7 ± 0.5 4.0 ± 0.6 8.7 ± 0.7 3.6 ± 0.6 3.1 ± 0.1 3.7 ± 0.4 6.3 ± 0.4

18: 2n-6 5.6 ± 0.2 5.8 ± 0.2 8.6 ± 0.3 8.6 ± 0.1 7.2 ± 0.2 5.3 ± 0.1 8.4 ± 0.1 8.4 ± 0.1 7.4 ± 0.220: 4n-6 1.2 ± 0.2 0.9 ± 0.0 0.9 ± 0.1 0.8 ± 0.1 0.5 ± 0.1 0.8 ± 0.0 0.8 ± 0.1 0.8 ± 0.0 0.4 ± 0.0

18: 3n-3 0.6 ± 0.1 1.1 ± 0.0 1.3 ± 0.1 1.2 ± 0.1 0.9 ± 0.1 1.0 ± 0.0 1.2 ± 0.0 1.2 ± 0.0 0.8 ± 0.018: 4n-3 0.9 ± 0.1 1.6 ± 0.1 1.3 ± 0.0 1.3 ± 0.1 1.6 ± 0.3 1.5 ± 0.1 1.3 ± 0.1 1.4 ± 0.1 1.5 ± 0.020: 4n-3 0.5 ± 0.0 0.9 ± 0.0 0.7 ± 0.0 0.7 ± 0.0 0.6 ± 0.1 0.7 ± 0.0 0.6 ± 0.0 0.7 ± 0.0 0.6 ± 0.020: 5n-3 (EPA） 5.3 ± 0.2 6.2 ± 0.5a 5.3 ± 0.4ab 4.8 ± 0.9ab 4.6 ± 1.1b 6.6 ± 0.2a 5.0 ± 0.3bc 5.1 ± 0.3b 4.7 ± 0.1c

22: 5n-3 (DPA） 2.2 ± 0.2 2.4 ± 0.1a 2.2 ± 0.2ab 2.1 ± 0.2b 1.5 ± 0.1c 2.4 ± 0.2a 2.0 ± 0.1b 2.1 ± 0.1b 1.4 ± 0.1c

22: 6n-3 (DHA） 11.4 ± 1.6 10.9a 11.3 ± 1.0a 9.4 ± 3.1a 4.9 ± 1.2b 11.4 ± 0.6a 10.2 ± 0.8a 10.5 ± 1.2a 5.1 ± 0.5b

Others 9.2 ± 2.8 5.9 ± 1.5 5.0 ± 0.3 5.2 ± 1.0 4.5 ± 2.4 8.9 ± 1.9 7.9 ± 2.0 8.5 ± 2.0 9.8 ± 0.2n-3 HUFA 20.3 ± 1.4 21.7 ±2.5a 20.9 ± 1.6a 18.3 ± 4.3ab 13.2 ± 2.9b 22.7 ± 0.6a 19.2 ± 1.2b 19.8 ± 1.5b 13.2 ± 0.6c

Mean ± standard deviation (n =5)．Values having different superscript letters are significantly different (P ＜0.05).Experimental diets are shown in Table 1.Fatty acid compositions are shown in Table 2.

Table 5. Proximate composition, phosphorus content, and fatty acid composition of muscle of yellowtail (n=5) fed theexperimental diets

Rearing season (days) Summer (90 days) Autumn (91 days)

Phosphorus content

Table 5.Proximatecomposition,phoshorusconntent,andfattyacidcompositionofmuscleinyellowtail(n=5)fedtheexperimentaldiets

47

Noriko ISHIDA, Tomohiko KOSHIISHI, Tatsuo TSUZAKI, Koji MAENO, Satoshi KATAYAMA, Minoru SATOH, and Shuichi SATOH

survival.Onlyslightlylessproteindigestioninporkmealbaseddietwas seen than theplantproteinbaseddiet.Andfeedingactivityonporkmealbaseddietswasactive.Fromtheseresults,porkmealwasthoughttobeavailableasasubstitutematerial.　Thebiggestproblemhasbeenshowntobe thereductionof feedingactivitywhenfishare fed lowfishmeal contentdiets (Aoki et al., 2000a, 2000b, 2000c). However, it is reported that an addedpalatability ingredients improved the feedingactivityineel(Takiiet al., 1984),yellowtail(Takiiet al., 1994),andtigerpuffer(Takiiet al., 1998).Inthepresentstudy,wefoundthatfishdidnotespeciallypreferplantproteintoanimalproteinandfishmeal.However, afterwashing the feed ingredients toremovetheextractivecomponent,thespittingratiodecreased inplant ingredient feedsand increasedinanimalprotein feeds,whichsuggested that theextractivecomponentsmightaffect feedingactivity(Niinumaet al., 2012,Yamauchiet al., 2012).Then,weaddedsomepalatabilityingredientstotheplantbaseddiet,fishswallowingratiowas increasedandspittingratiowasdecreased.Fromtheseresults,weused theskipjackpeptide (SP)and fishsauce (FS)aspalatability ingredients inorder to improve thefeedingactivity. In theresults,higherpalatabilitywasseenwhenusingbothingredients.Thebiggestproblemofreductionof feedingactivity innon-fishmealdietswasthoughttobealmostresolvedbytheadditionofpalatabilityingredients.　Themeatqualityofyellowtail fedwithnon-fishmealdietswerecomparedtocontrolyellowtailbyapaneltest,whichshowedadifferenceincolorandsmell.Thenweused colorimeter andElectronicNoseSystemofAlphaMOS(http://www.alpha-mos.co.jp/products/sensory.html).The fish fedbynon-fishmealdietshadwhitermuscleanda less fishysmell (Tohataet al., 2013).Now, theeffectofnon-fishmealdietsonfishmeatqualityandasamethodof enhancing themeat quality control is beingdetermined. 　Asdescribedabove,wealternated fishmeal toplantoranimalproteinmaterials tomakenon-fishmealdiets,andfedthemtooneyearoldyellowtail,andgotsimilargrowth,bodyandbloodcompositions.Themainreason for thesimilargrowthrateswasthoughttobetheadditionofpalatabilityingredients

and enzyme mixtures (Table 1) . As we usedbothplant and animalmaterials,we suggest thepossibilityofmanymaterialsfornon-fishmealdiets.Morestudyisneededtoimprovelowcostdietsandproducehigherqualitymeat.　This work was supported by Research andDevelopmentProjects forapplication inpromotingnewpolicyofagriculture,forestryandfisheriesfromtheMinistryofAgriculture,ForestryandFisheriesofJapan.

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