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Conditioning of broodstock of tiger grouper, epinephelus fuscoguttatus, in a recirculating aquaculture system

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Introduction Closingthecycleofcommerciallyexploitedfishina recircu-latingaquaculturesystemisgrowinginimportanceforavariety ofreasonsthatincludeovercomingthedifficultiesingettinglive broods

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j ou rn a l h o m e p a g e :w w w e l s e v i e r c o m / l o c a t e / a q r e p

Saleem Mustafa∗, Mohd Hafizzie Hajini, Shigeharu Senoo, Annita Yong Seok Kian

Borneo Marine Research Institute, Universiti Malaysia Sabah, Kota Kinabalu, Sabah 88400, Malaysia

a r t i c l e i n f o

Article history:

Received 9 February 2015

Received in revised form 5 August 2015

Accepted 18 September 2015

Available online 28 September 2015

Keywords:

Tiger grouper

Broodstock

Conditioning

Socio-demographic cues

Intrinsic factors

Sex reversal

a b s t r a c t

1 Introduction

Closingthecycleofcommerciallyexploitedfishina

recircu-latingaquaculturesystemisgrowinginimportanceforavariety

ofreasonsthatincludeovercomingthedifficultiesingettinglive

broodstockfromthewild, exorbitantcost,biosecurityproblems

andimpactonthemarineecosystem

Inahatchery,closingthelifecycleoftigergrouper(Epinephelus

fuscoguttatus)thatrequiresseveralyearstomatureandis

protog-ynoushermaphrodite,presentssomechallenges.However,since

adequatesupply of high-qualityseedof this species isa major

constraintfacedbytheaquacultureindustry,especiallythe

small-and-mediumenterprises,thesechallengeshavetobeaddressed

Seedqualitydependsheavilyonbroodstockcondition.While

temperatureandphotoperiodarethetwomainenvironmentalcues

thatcontrolthereproductivecycle(Sudaryantoetal.,2004)other

factorsincludingthenutritionalstatus,and parameterssuchas

salinityanddissolvedoxygendoinfluencethephysiological

con-ditionofthefish(Simetal.,2005;Sugamaetal.,2012).Anideal

∗ Corresponding author Fax: +60 88 320261.

E-mail addresses: saleemacademic@gmail.com , Saleem@ums.edu.my

(S Mustafa).

broodstockmanagementenvisagesmimickingtheconditionsthat thefishfacesinitsnaturalenvironment.Becausetheselected spec-imensofthefishwereproducedinthehatchery,theyarealready accustomedtocultureconditionsandthereforeeasiertodevelop intobroodstockcomparedtotheirwildcounterpartsorparentsof thiscohortsourcedfromthewildpopulations

Thisstudywasundertakenfordeterminingtheoptimum con-ditionsfordevelopingtigergrouperbroodstockbyenvironmental controlsaimedat stress reduction andbalanced nutrition Pro-ductionofseedbyspawningoftheenvironmentallyconditioned captivebroodstockhasmanyadvantagesoverspawninginduced

byhormoneinjection.Hormonetreatmentcausesstressof han-dling,injection,and/orimplantationofexogenoussubstances,and producessideeffectsaswell.Thispaperpresentsdataonthe posi-tiveeffectsofeffectivelycontrollingbroodstockrearingconditions forgrowthanddevelopmentofgonads

2 Materials and methods

ThisexperimentwascarriedoutatthefinfishhatcheryofBorneo Marine ResearchInstitute, UniversitiMalaysia Sabah,Kota Kin-abalu.Broodstockareaofthehatcherywascoveredwitharoof butthesideswereopen,allowinglighttoenter.Noartificiallight wasusedtomanipulatethephotoperiod.Eachtankwasof150m3 http://dx.doi.org/10.1016/j.aqrep.2015.09.004

2352-5134/© 2015 The Authors Published by Elsevier B.V This is an open access article under the CC BY-NC-ND license ( http://creativecommons.org/licenses/by-nc-nd/4.

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capacity,roundin shapeand medium-rangeblueincolor.Tank

of this volume is consideredideal for broodstockmanagement

inthehatcheries Itprovidesadequatespaceforswimmingand

specificcourtshipbehavior.Thisisconsistentwiththesuggestion

ofSugamaetal.(2012)regardingthetanksize,shapeandcolor

(preferablymedium-rangeblue,greenorgraycolorandavoiding

shadesthatareeitherverylightorverydark)

Specimensofthetigergrouperwhichvisiblylookednormalin

bodyshapeandcolor,appearedhealthythroughgeneralactivity

(swimming,feedingandquickresponsetoexternalstimuli)and

whichweredevoidofanyskeletalabnormalitiesorexternalsignsof

infectionorinjurywereselectedaspotentialbreedersforthetrials

Thesespecimenswereproducedinthesamehatcherybyselective

breedingoffounderstockthat originatedfromwildpopulation

Thetestspecimens(broodstockcandidates)belongedtothefirst

generationofthehatchery-producedfish.Sixty-eightspecimens

oftigergrouperofaveragetotallength=65.2±11.2cmandbody

weight=5601±2699gwererearedfortrialsconductedin

brood-stocktanks.Fishwereequallydividedinthetwotanks.Maintaining

waterqualityconditionsinsuchlargetanksischallengingbut

man-ageablegiventheadequatesupplyoffilteredseawater,aeration

andairliftpumps.Tankswereprovidedwithwaterrecirculation

systemandthefishwereobservedthroughsidewindows.Tanks

receivedwaterfilteredbydynasandfiltrationsystemthatensured

removalof solids.Nitrogenous wastes inthe formof ammonia

andnitritewerecontrolledbybiofiltersinanadjoiningtankthat

containedsubstratesforcolonizationofnitrifyingbacteria

(Nitro-somonasandNitrobacter).Aeratorswereusedtohelpincirculation

ofdissolvedoxygenandeliminationofnitrogenouswaste

Tem-perature,salinity,pH,dissolvedoxygen,nitriteandammoniawere

routinelymonitored

Thefishspecimenswereofferedfeedcontainingpreyfishthrice

aweekattherateof3%bodyweightwhichprovidedabout50%

protein(dryweightbasis).Thistreatmentofthefishcontinuedfor

6monthsduringwhichtheirgrowth,somaticconditionandsigns

ofsexdifferentiationwererecorded.Fishweregenerallyobserved

daily,particularlyatthetimeoffeedingbutmeasurementswere

takenonaweeklybasis.Asamatteroffact,theexperimentstarted

withthesame-sexspecimensofthisprotogynoushermaphrodite

fish.Forthespecificpurposeofnoticingsexdifferentiation,

obser-vationsonanychangeinthebehaviorwerecarriedout

Length–weightrelationshipwasestablishedbyusingthe

stan-dardallometricequation:W=aLb,where,W=weight(g),L=total

length(cm),a=constant(interceptinthegraph)andb=exponent

(slopein thegraph).Thisequationis for anon-linear situation

which does not offer a direct solution for interpretation of ‘a’

and‘b’.Itwas,therefore,logarithmicallytransformedforalinear

regressionmodel:logW=loga+alogL,whereloga=constantand

b=exponent.Thisequationcanbeusedforpredictingthelogarithm

ofweightasafunctionofthelogarithmoflength

Conditionfactor(K)wascalculatedbytheformula:K=100W/L3,

where,W=bodyweightofthefish(g)andL=totallength(cm)

3 Results

achieve water quality parameters in the tanks in the range:

salinity=29.7–31.0‰, temperature=26.4–28.9◦C, dissolved

oxy-gen=5.8–6.4ppm and pH 7.2–7.5 The nitrogen-nitrite and

unionized ammonia never exceeded 0.05ppm and 0.02ppm,

respectively

Analysisof length–weightregression produced theformula:

logW=−1.5855+2.9185logL(Fig.1).Thecorrelationcoefficient,

R2(0.9501)wassignificantlyhigh(P<0.005),suggestingasteady

progressionofthetwogrowthparameters.Theexponent(b)value

y = 2 9185x - 1 5855 R² = 0 9609

3.00 3.20 3.40 3.60 3.80 4.00 4.20

1.65 1.70 1.75 1.80 1.85 1.90 1.95 2.00

Log L (cm) Length Weight Rel ationship

Fig 1. Length–weight relationship in tiger grouper, Epinephelus fuscoguttatus.

of2.9185indicatednomajordeparturefromthecube-law relation-shipbetweenlengthandweightalthoughtechnicallyspeakingany valuelessthanthree(>3)characterizesnegativeallometricgrowth thatimpliesthatthefishbodyprofilebecameslightlymoreslender

asitgrew.Theconditionfactor(average=1.86)waswithinthe nor-malrangeforahealthytigergrouper,reflectingthatthefishwerein

agoodsomaticconditionasaresultofthefavorableenvironmental conditionsandappropriatefeedingregime

Thestockedfishstartedwithfemalesexbut5outof68 speci-mensshowedmale-likebehavior

4 Discussion

Itisevidentfromthegrowthexponentinthelength–weight relationshipandtheconditionfactorof thegrouperstocksthat thehatcheryprovidedsuitablecultureenvironment.Thisaugurs wellforbroodstockmanagement,especiallygrowth,gonad devel-opment,fecundity,eggqualityand exercisingcontrolontiming

ofmaturationandspawning.Itisnotuncommonforfishreared

inhatcheriestosufferfromreproductivedysfunctionandlossof fertilityifcaptivityconditionsarenotproperlymaintained.Tank size, shape and colorare important in maintainingbroodstock forextended periodsin thehatchery Adequatespacefor large-sizedfishlikegrouperisnecessaryespeciallyforcourtshipthatin thisspeciesisnecessaryforbreeding.Whilediscussingthistopic Benetti(2002)hasemphasizedthatthelargerthefish-holdingarea, thebetteritisforthefish.Tanksof150-toncapacityusedinthis trialtogetherwitheffectivecontrolonwaterqualityandnutrition wereamongthemaincontributingfactorsforthewellbeingofthe captivestocks

Beingaprotogynoushermaphroditethetigergrouperstartsits earlyphaseoflifeasafemaleandatalateragesomespecimens changesextobecomemale(Pearsetal., 2007).It is,therefore, expectedthatfemalesoutnumberthemalesandthesexratioto

behighlyskewedtowardthefemalesexevenafterdifferentiation

ofafractionofthepopulationintomale.Therearemanyviewson whattriggerssexreversalbutitisunderstoodthatthefactorsmay

beinternal(intrinsic)orexternal(extrinsic).Inthenatural environ-ment,sexchangeingroupersisknowntooccurduringspawning aggregationforreproductivesuccessandifatthattimethescarcity

ofmalesbecomesalimitingfactor,somefemalesswitchoverto malesextoensurethatbreedingandpopulationrecruitmenttake place.Generally,thedominantfemalesundergothissortofsex con-version.Adifferentsituationprevailsinthehatcherytanks.The socialandenvironmentalcuesinthehatcheryaredifferent.Also, thereisnoaggregationdrivenbynaturalinstinctandurgetomate similartowhatprevailsinnature.Fishmovingoverlongdistances

intheirnaturalenvironmentforaggregationwiththepurposeto matewillcertainlybemorevigorousinthisactivityandtheir hor-monalturnoverwillbequantitativelydifferentfromstocksheld

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restrictedarea,whileinnaturetheydonotliveingroups.Obviously,

thereissomesocialinteractionasamatterofroutinewhileliving

overextendedperiodsandthisperhapscandiminishtheintensity

ofmatingandreproductiveactivityseeninthewild.Thedesireto

mateisinstinctiveandmotivatedbyphysiologicalfactorsbutsocial

andenvironmentalfactorshaverolestoplay.Theexactnatureof

thecomplexcuesandtheleveloftheirinfluenceonsexualityare

difficulttounderstand

Thetestspecimensstockedintanksweremixedagegroups

Theywereallfemalesduetoprotogynousconditionofthisfish

Evenwhennofunctionalmalewasintroduced,atendencyin a

smallnumberofthesespecimenstoturnintomalewasnoticed

Whenafunctionalmalewasintroduced,itdidnotseemtoproduce

anyeffectonsextransformationasthenumberdidnotchange

ThisviewisatvariancewiththatofSugamaetal.(2012)who

sug-gestedthatthepresenceoffunctionalmalefishcouldrepresssex

changebythefemale.Itislikelythatafunctionalmaledoesnot

makeanysignificantimpactwhendominantfemaleshavealready

startedtransitioningtomalesex.Changeinthesexthatoccurredin

5ofthe68specimenexaminedindicatedthatevenintheabsence

ofthemalesthecuesareatworktotriggerthelargersizedfish

ofmatureagetobecomemale.Thiscouldbeattributedto

socio-demographiccuethatisanexternalfactor.Sugamaetal.(2012)

havedocumentedthatsexchangeintigergrouperinbroodstock

tanksissociallymediated.Totheextentthatthesexreversalonly

involvedfishof4–5yearsofage,itseemstobeage(orsize)-related

whichisendogenouslycontrolled.Youngerfishof2–3yearsofage

showednoevidenceofsexreversal,soapparentlytheperception

ofsocialcuesdependsonageorinternal(physiological)condition

ofthefish

Signsofsexdifferentiationobservedwereintheformofchange

inbehavior.Thisincludedonsetofmale-likebehavior,increased

patrollingoftheentiretank,shakingofheadandvigorous

swim-mingwheninclosevicinityoffemaleconspecifics.Itisexpected

thatthischangeinbehaviorisundertheinfluenceofincreasein

malesexhormones.While behavioralchangecanhappenmuch earlierthanthecompletetransitionofthegonadfromovaryto testis as it involves modification of gonad morphology and its steroidogeniccapacity.Thefishcanbecomeatrulyfunctionalmale uponcompletionofthisprocess.Howlongittakestoachievethis stageisaninterestingtopictopursue

Probably,thisisthefirstreportofitskindontigergrouperthat providesaconvincingexplanationoftheroleofbothinternaland externalfactorsinsexdifferentiationintigergrouperinthe hatch-ery tanks.Attainingfunctionalfemaleand malestatusby these specimenswilldefinethesuccessoftheclosedcycleaquaculture

oftigergrouper

Acknowledgement

ThisstudywasfundedbytheMinistryofEducationofMalaysia undertheHigherInstitutions’CenterofExcellence(HICoE) pro-gram

References

Benetti, D.D., 2002 Advanced conditioning systems for marine fish broodstock Glob Aquacult Advocate, 22–23.

Pears, R.J., Choat, J.H., Mapstone, B.D., Begg, G.A., 2007 Reproductive biology of a large aggregation-spawning serranid, Epinephelus fuscguttatus (Forskål): management implocation J Fish Biol 71, 795–817.

Sim, S.Y., Rimmer, M.A., Toledo, J.D., Sugama, S., Rumengan, I., Williams, K.C., Phillips, M.J., 2005 A Guide to Small-Scale Marine Finfish Hatchery Technology NACA, Bangkok, Thailand, 17 pp.

Sudaryanto, Meyer, T., Mous, P.J 2004 Natural spawning of three species of grouper in floating cages at a pilot broodstock facility at Komodo, Flores, Indonesia SPC Live Reef Fish Information Bulletin 12 Noumea Cedex, New Caledonia.

Sugama, K., Rimmer, M.A., Ismi, S., Koesharyani, I., Suwirya, K., Giri, N.A., Alava, V.R., 2012 Hatchery Management of Tiger Grouper (Epinephelus fuscoguttatus):

A Best-Practice Manual ACIAR Monograph No 149 Australian Centre for International Agricultural Research, Canberra, 66 pp.

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