North American Journal of Aquaculture Relation between Nucleic Acid Indices and Growth Rate in ...

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Relation between Nucleic Acid Indices and Growth Rate
in Fed and Fasting Juvenile Scup
Renee Mercaldo-Allen a , Catherine A. Kuropat a , Dean M. Perry a & Dylan H. Redman a
a National Oceanic and Atmospheric Administration, National Marine Fisheries Service,
Northeast Fisheries Science Center , Milford Laboratory , Milford , Connecticut , 06460 , USA
Published online: 23 Jan 2014.
To cite this article: Renee Mercaldo-Allen , Catherine A. Kuropat , Dean M. Perry & Dylan H. Redman (2014) Relation between
Nucleic Acid Indices and Growth Rate in Fed and Fasting Juvenile Scup, North American Journal of Aquaculture, 76:1, 1-8,
DOI: 10.1080/15222055.2013.829145
To link to this article: http://dx.doi.org/10.1080/15222055.2013.829145
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NorthAmericanJournalofAquaculture76:1–8,2014
AmericanFisheriesSociety2014
ISSN:1522-2055print/1548-8454online
DOI:10.1080/15222055.2013.829145
ARTICLE
Relation between Nucleic Acid Indices and Growth Rate
in Fed and Fasting Juvenile Scup
ReneeMercaldo-Allen,*CatherineA.Kuropat,DeanM.Perry,
andDylan H.Redman
NationalOceanicandAtmosphericAdministration,NationalMarineFisheriesService,
NortheastFisheriesScienceCenter,MilfordLaboratory,Milford,Connecticut06460,USA
4
1
0
2
y
a Abstract
M
0 Therelationshipbetweennucleicacidindicesanddailyinstantaneousgrowthratewasdeterminedforjuvenile
2 ScupStenotomuschrysopsinthelaboratory.Weight-basedgrowthofScupwasmeasuredforthree1-weekintervals
45 duringa6-weekfeedingexperimentandshowedasignificantpositivecorrelationwithbothRNAconcentration(RNA;
0: µg/mgtissuewetweight)andDNAconcentration(DNA;µg/mgtissuewetweight),whiletheRNA:DNAratiowasnot
2
at significantlycorrelatedwiththeweight-orlength-basedgrowthrate.TheconcentrationofDNAshowedasignificant
] negativecorrelationwithlength-basedgrowthrate.Meanfishweightincreasedfrom1.022to10.404g,andmeanTL
s
e increasedfrom43.219to85.377mm.Changesinnucleicacidindicesweremeasuredinfishthatweredeprivedof
eri foodovera9-dperiod.TheRNAandRNA:DNAratiovaluesdeclinedsignificantlywithin5dafterfastingbegan,
h
s whileDNAremainedrelativelystable.Thisstudyprovidesbaselinenucleicacidvaluesforfedandstarvedjuvenile
Fi
f Scupoverarangeoffishsizes.
O
nt
e
m
rt
a
p
De TheScupStenotomuschrysopsisfoundinLongIslandSound theRNA:DNAratio(RD),havebeenwidelyappliedasrelative
y [ andalongtheeasternseaboardfromCapeCod,Massachusetts, measuresofrecentgrowthornutritionalconditioninthelarvae
b
d toCapeHatteras,NorthCarolina(Morse1978),andthisspecies andjuvenilesofmanymarinefishspecies(FerronandLeggett
de has been identified as a candidate for aquaculture production. 1994;Bergeron1997;Ch´ıcharoandCh´ıcharo2008).
a
o Juvenile Scup have a large gape size, can be weaned to a pel- Laboratoryandfieldstudiesdocumentastrongrelationship
nl
w letedfeedrelativelyquickly,anddemonstratehighsurvivaland between RD and weight-based growth in juveniles of numer-
o
D rapidgrowthoncommercialdietsinthelaboratory(Perryetal. ous fish species (e.g., Varnavskiy et al. 1991; Mathers et al.
2009). Appraisal of the Scup as an aquaculture species re- 1992; Foster et al. 1993; Malloy et al. 1996; Rooker and Holt
quiresinvestigationinto(1)culturestrategiesforoptimalgrowth 1996; Rooker et al. 1997; Peck et al. 2003; Mercaldo-Allen
andsurvivaloffishinresearchandcommercialoperationsand et al. 2008; Edwards 2009). The RNA-based indices respond
(2)methodsforassessingtheconditionofculturedfish. rapidly to changes in protein synthesis, are highly sensitive to
Nucleicacidindicatorscanbeusedinaquacultureasatool variations in both weight- and length-specific growth, and are
for determining the effects of variable diet formulations and advantageousbecausetheyestimategrowthforadiscretepoint
ration amounts on fish growth and physiology (Carter et al. in time based on a single sampling, independent of the need
1998;Buckleyetal.1999;Waltheretal.2010).Concentrations for repeated morphometric measurements (Arndt et al. 1996;
of RNA in the white muscle tissue of young fish correspond Carter et al.1998). Biochemical measures are highly sensitive
to changes in protein synthesis and somatic growth, whereas tovariationinfoodavailabilityandrespondwithindayswhen
DNAconcentrations(thecellnumberwithinagivenweightof food is withheld (Ferron and Leggett 1994; Edwards 2009).
tissue)remainrelativelyconstant.Nucleicacidindices,suchas Validationofthenucleicacid–growthrelationshipforparticular
*Correspondingauthor:[email protected]
ReceivedMarch25,2013;acceptedJuly22,2013
1
2 MERCALDO-ALLENETAL.
species,bodysizes,andlifestagesofinterestisrequired(Peck determined to the nearest 0.01 g, and initial length (ILN) was
etal.2003)beforetheseindicescanbeusedforevaluatingthe measuredtothenearestmillimeterTL.Oneweeklater(weeks
performanceofcommercialfeeds,identifyingoptimalstocking 2,4,and6),finalweightandTLofeachtaggedfishweremea-
densities,ormonitoringthequalityofholdingconditionsforfish sured, and the fish were frozen whole at −80◦C for later RD
inanaquaculturesetting.Therelationshipbetweennucleicacid analysis. The IWT, ILN, final weight, and final length values
indicesandgrowthofjuvenileScuphasnotyetbeendescribed. were used to calculate the daily weight- and length-based in-
Weconductedastudytoexaminetherelationshipsbetween stantaneous growth rates (i.e., GWT and GLN) of tagged fish
nucleic acid measures (RD, RNA concentration [RNA], and ineachoftheeighttanksduringthethreeintervals(week1–2:
DNA concentration [DNA]) and instantaneous growth rate in 7 d; week 3–4: 7 d; and week 5–6: 6 d). The GWT (g/d) was
weight (GWT) or length (GLN) in fed juvenile Scup at three calculatedas[log (finalweight)−log (IWT)]/(numberofdays
e e
differentsizeintervalsduringtheirfirstsummer.Changesinnu- in the interval), and the GLN (mm/d) was calculated as (final
cleicacidconcentrationswerealsomeasuredinScupthatwere length−ILN)/(numberofdaysintheinterval).
deprived of food over a 9-d period. The present results pro- Since tagged fish were not uniquely identified, individuals
videbaselinenucleicacidvaluesfordescribingfedandstarved could not be followed, so a mean growth rate calculated for
conditionofjuvenileScup. each group of 5 fish/experimental tank was used in statistical
analyses. Sample size was eight tanks for weeks 2 and 6 and
seventanksforweek4(onetankwasomittedduetoinsufficient
4
1 METHODS data),yieldingatotalof23tankmeans.
0
y 2 Experimentaldesign:feedingstudy.—DuringlateJuly2008, Experimental design: food deprivation study.—Twenty fish
Ma age-0 Scup were collected near Milford, Connecticut, aboard were weighed, measured, and frozen for nucleic acid analysis
0 the National Oceanic and Atmospheric Administration vessel todetermineinitialsizeandconditionoffield-collectedfish.To
2
5 R/VVictorLoosanoff;thefishwerecapturedbyusinganotter followchangesinnucleicacidconcentrationduringfooddepri-
4
0: trawl lined with knotted twine (1.61-cm square) mesh. Scup vation, 45 fish from the same cohort (group) were placed into
2
at wereheldinaeratedcoolers,transportedtothelaboratory,and eachoftwotanksandwereheldunderthecultureconditionsde-
] acclimatedtoholdingconditionsfor1–2weekspriortotheex- scribedearlier,exceptthatnofeedwasadministered.Sampling
s
rie periments. Forty-five fish were placed into each of eight 76-L began after 24 h, and five fish were sacrificed daily from each
e
h aquarium tanks that received flow-through, sand-filtered sea- tankovera9-dperioduntilnofishremained.Sampledfishwere
s
Fi water at ambient temperature and a flow rate of 1.5 L/min per weighed,measured,labeledindividually,andfrozenat−80◦C
f
O replicate.Aquariawereheldwithinalargetankthatservedas for subsequent analysis. Daily sample size was 10 fish except
nt awaterbathtomaintaintemperature.Thisexperimentwaspart on day 9, when nine fish were sampled. The food deprivation
e
m of a larger study to assess the performance of two different experimentwasconductedduringJuly31–August8,2008.
rt
a commercialfeeds(Perryetal.2013).Fourtanksreceivedcom- Biochemical analyses.—To obtain white muscle tissue for
p
e
D mercial diet 1 (55.5–58.5% protein, 13.4–14.5% lipid), while analysis, frozen fish were dissected on a tray set on ice. The
[
y theotherfourtankswerefedcommercialdiet2(59%protein, skin was discarded, and the front portion of the fillet was re-
b
d 16% lipid). Adjacent tanks received different feeds. Fish were moved from one side. Tissue samples dissected for RD analy-
e
d fed to satiation, and diets were hand fed to the fish four times sis varied with fish size and ranged from 0.105 to 0.333 g for
a
nlo daily at an initial ration of 12% of body weight per day. As equivalentportions.Dissectingtoolswererinsedwithdeionized
w pelletsizeincreased,feedrationwasgraduallydecreasedto5% waterbetweendissectionstoavoidcontamination.Eachtissue
o
D ofbodyweightperday.Forbothdiets,pelletsizechangedover sample was weighed to the nearest 0.001 g and was placed in
theexperimentalperiodasfollows:fishwerefed200–400-µm a test tube within an ice slurry bath. The tissue was homoge-
pelletsduringweek1;400–600-µmpelletsduringweek2;500– nized in ice-cold distilled water by using a Janke and Kunkel
900-µmpelletsduringweeks3and4;and0.8–1.0-mmpellets Ultra-Turrax tissue homogenizer. Replicate aliquots were im-
duringweeks5and6.Seawatertemperature,salinity,anddis- mediately frozen, and biochemical analysis of the tissues was
solved oxygen readings were measured daily (YSI Model 85; completedwithin48hoffreezing.
YellowSpringsInstruments,Inc.,YellowSprings,Ohio).Light Muscletissuesampleswereanalyzedbyusinganultraviolet-
intensitywasmaintainedat1,000lx,withaphotoperiodof9h basedmethodaccordingtoBuckleyandBulow(1987)asmod-
light:15hdark.Thefeedingexperimentwasconductedduring ified by Kuropat et al. (2002). First, free nucleotides were re-
July31–September9,2008. movedviaaseriesofwasheswithcoldperchloricacid(HClO ).
4
Growth of juvenile Scup was measured over three distinct TheRNAwasthenhydrolyzedwithpotassiumhydroxide,and
1-week intervals to track growth rates and nucleic acid con- the hydrolysate was acidified with cold HClO to remove the
4
centrations over a 6-week period. At the start of each interval RNA from the DNA and protein. The DNA was then both
(weeks1,3,and5),asubsetoffivefishwasremovedfromeach hydrolyzed and separated from the remaining protein by the
aquarium and those fish were tagged in the dorsal region with addition of hot HClO . Both RNA and DNA were estimated
4
injectedelastomer.Initialwetweight(IWT)oftaggedfishwas fromtheabsorbanceofthehydrolysateat260nmbasedonan
NUCLEICACIDINDICESANDGROWTHOFJUVENILESCUP 3
extinction coefficient of 0.03 (for a 1-µg/mL solution of hy- 12
drolyzed RNA or DNA). Absorbance was measured using a 11 initial Weight
PerkinElmerUV/VISLambda35spectrophotometer.Concen- 10 final
trations of RNA and DNA were calculated and expressed as 9
microgramspermilligramoftissuewetweight.Duetosmaller (g) 8
initialfishsizes,replicatesampleswereavailablefor22%offish ht 7
g
inthestarvationstudyascomparedwith100%offishinthefeed- ei 6
W
ingstudy.Whensufficientwhitemuscletissuewasavailablefor n 5
a
replicatealiquots,thevalueswereaveraged.Asaqualitycontrol e 4
M
measure, a large quantity of adult Scup muscle tissue was ho- 3
mogenizedandfrozenin0.2-galiquots.Onecontrolsamplewas 2
processedduringeachanalysisalongwiththetissuesamplesto 1
verifytheaccuracyoftherun(Buckleyetal.1999;Caldarone 0
et al. 2006). Analytical results were accepted if the measured 2 weeks 4 weeks 6 weeks
controlvalueswerewithin10%ofestablishedvalues.
Statistical analysis.—Data sets were tested for normality 90
Length
4 prior to analysis. When data did not meet the assumption of 85 initial
1 normality,nonparametricstatisticaltestswereused.Spearman’s final
0 80
May 2 rtwanekenornduecrlceoicrraeclaidtiomnewasausreusse(dRNtoAd,eDteNrmAi,naentdheRaDss)o,ciniaittiiaolnsbizee- mm) 75
0 (ILNandIWT),andgrowthrates(GWTandGLN)duringthe h ( 70
s] at 20:45 2 fwKeereurFdesioknbragatlhs–eeeWxdfpaooelonrlidismtdaoenennpketr-.miwvSeaatatayinotAisns.tNsictOuadVlyAa,nooannlyer-sawensakyfsoA(rfoNtrhOReVNfAeAe(dfaionnrdgRDsDtNu)Adoy)r Mean Lengt 566505
rie was used to test for significant differences between aquaria. 50
e
h Dunn’smethodwasusedtocomparenucleicacidconcentrations
s 45
Fi on each day of the starvation trial with the initial values for
f 40
O fed fish. Statistical analyses were performed with SigmaStat
nt version3.5. 2 weeks 4 weeks 6 weeks
e
m
art FjuIvGeUniRleES1c.upMateeaanch(±oSfDth)reineigtiraolwatnhdifinntearlvwalesi(gwhtese(kg2):aNnd=le8n;gtwhese(km6m:NTL=)o8f;
p RESULTS
e week4:N=7).
D
[
y FeedingExperiment
b
d Scup grew rapidly on both pelleted diets, increasing sub-
e werelow:onefishwasunaccountedforintank5duringweek
d stantiallyinweightandlengthafter6weeksofconsumingthe
wnloa fveeendilse. MSceuapnaIrWe dTi,spILlaNy,edfininalFwigeuirgeht1,;atnhdesfienraelsuleltnsgsthhofworthjua-t 2te,manpdertahtuisrew±asSmDoswtalisk2el3y.4du±e0to.7t2a◦gClo(sras.ngTehe=m21ea.8n–s2e4a.w9◦aCte)r,
Do fishdoubledinweightandlengthfromthebeginningtotheend salinityaveraged25.3 ± 0.69psu(range=23.1–26.0psu),and
mean dissolved oxygen concentration was 5.87 ± 0.43 mg/L
oftheexperiment.Overthe6weeks,meanGWTrangedfrom
(range=4.65–7.24mg/L)overthe6-weekexperiment.
0.041 to 0.084 g/d and GLN ranged from 0.829 to 1.6 mm/d.
Moreinformationconcerninggrowth,feedconversionrates,and
dietcompositionoveralongerperiodisdetailedbyPerryetal. FoodDeprivationExperiment
(2013).Basedont-tests,nucleicacidindicesofthegroupsgiven Duringthefooddeprivationexperiment,nucleicacidvalues
thetwofeedsdidnotsignificantlydifferduringanyofthethree didnotdiffersignificantlybetweenthetwoaquaria(P>0.050);
growthintervals,sodataforthetwodietgroupswerecombined. therefore,thedatafortheaquariawerecombined.TheRDand
Both RNA and DNA exhibited significant positive correla- RNA of starved fish declined over the 9-d period. According
tionswithGWTinfedjuvenileScup(Table1).Theconcentra- toDunn’stest,RDandRNAdifferedsignificantlyfrominitial
tion of DNA had a significant negative correlation with GLN. valuesafterfishhadfastedfor5d(Figure3).Theconcentration
Both RNA and DNA were significantly negatively correlated of DNA did not differ significantly from initial values at any
withIWTandILN.TheRDwasnotcorrelatedwithGWT,GLN, time. There was no evidence of mortality until the final day
IWT, or ILN. The relationships between (1) RNA and GWT, ofsampling(day9),whenonefishremainedunaccountedfor.
(2) DNA and GWT, and (3) DNA and GLN for each growth Meanseawatertemperature ± SDwas23.6 ± 0.60◦C(range=
intervalareplottedinFigure2.MortalityratesoftaggedScup 22.3–24.5◦C), mean salinity was 25.5 ± 0.28 psu (range =
4 MERCALDO-ALLENETAL.
TABLE 1. Spearman’srankordercorrelationcoefficientsforthejuvenileScupfeedingexperiment.Variablesincludenucleicacidindices(RNA:DNAratio
[RD];RNAconcentration[RNA],µg/mgtissuewetweight;DNAconcentration[DNA],µg/mgtissuewetweight),instantaneousgrowthrates(inweight[GWT]
andlength[GLN]),andfishsize(initialweight[IWT],g;initiallength[ILN],mmTL).TheP-values(inparentheses)lessthan0.05indicatesignificance.Analyses
werebasedontankmeans(N=23).
Variable RD RNA DNA
RNA −0.0642(0.767)
DNA −0.6250(0.001) 0.738(<0.0001)
GWT −0.0494(0.820) 0.757(<0.0001) 0.613(0.0019)
GLN 0.3260(0.127) −0.239(0.268) −0.499(0.0157)
IWT 0.1420(0.512) −0.812(<0.0001) −0.765(<0.0001)
ILN 0.0930(0.669) −0.830(<0.0001) −0.746(<0.0001)
24.9–25.9 psu), and dissolved oxygen concentration averaged growth activity, whereas in older juveniles protein synthesis
5.98 ± 0.26mg/L(range=5.4–6.39mg/L). is directed toward a broader range of processes, including the
manufactureofenzymesandincreasedcellturnover(Caldarone
14 DISCUSSION etal.2006).Thisshiftinenergystorageandmetabolismaccom-
0
2 paniesadecreaseinproteinrequirementsamonglargerfishas
y
a EvaluatingtheRelationshipsbetweenNucleicAcid theyapproachmaturity(FrommelandClemmesen2009).Mea-
M
0 IndicesandGrowthofFedScup surement of lipid or fatty acid content in juveniles of various
5 2 In juvenile Scup that received feed, we observed a decline sizesmightclarifytheroleofchangingbodycompositioninthe
4 in DNA that corresponded to an increase in GLN, a slowing RD–growthrelationship(Vasconcelosetal.2009).Ourresults
0:
2 of GWT, and an overall increase in body size across the three suggest that RD should not be broadly applied as a condition
] at measurement intervals. Variation in DNA over time may re- measure in juvenile fish, even within a given species, without
s
e flect an alteration in growth processes among fish of rapidly first validating the RD–growth relationship for the range of
ri
he increasingbodysize.Hypertrophy(anincreaseincellsize)ac- sizes to be compared (Buckley et al. 1999), since shifts in
s
Fi countsformuchofthemusclegrowthamonglargerjuvenilefish physiologyandmorphologycanoccuroverthejuvenileperiod.
Of (Arndtetal.1996;Ch´ıcharoandCh´ıcharo2008;Frommeland ThestrongrelationshipweobservedbetweenRNAandGWT
nt Clemmesen2009).Ascellsizeincreases,thequantityofDNA has also been documented in juveniles of other fish species
e
m inagivensampleofmuscletissueisdiluted,thusloweringthe (e.g.,Fosteretal.1993;Mathersetal.1993;Carteretal.1998;
art DNA concentration (Love 1970). The positive correlation be- Kuropat et al. 2002; Mercaldo-Allen et al. 2008; Ciotti et al.
p
e tweenDNAandGWTobservedinthepresentstudydiffersfrom 2010). In our study, GLN was not closely aligned with RNA,
D
y [ the results of previous studies, in which a negative correlation although an increase in GLN did occur over the three inter-
d b betweenDNAandweight-basedgrowthwasfoundforsmaller valsdespiteadeclineinRNA.InvolvementofRNAinprotein
de andyoungerjuvenilesofothermarinespecies(Mercaldo-Allen production—generallyaccompaniedbyaccretionofmuscletis-
a
o et al. 2006, 2008; Edwards 2009). The response of DNA to sueandincreasedfishweight—mightexplainwhytherelation-
nl
w reductionsinGWTmaynotbeconsistentacrossbodysizesor ship with RNA was stronger for GWT than for GLN. As fish
o
D ontogeneticstagesinyoungfish.Measurementsinnewlysettled transitiontowardadulthood,growthmayshiftfromafocuson
Scuparenecessaryfordeterminingwhetherthestrengthanddi- weightgaintoafocusonlengtheningofbodymorphology.Al-
rectionoftheDNA–GWTcorrelationchangeoverthecourseof though our Scup were fed to satiation, RNA and GWT both
ontogenyinthisspecies. declined across the three measurement intervals. Juvenile fish
Our feeding study found no relationship between RD and may continue to increase in body size despite reductions in
GWTorGLNofjuvenileScup;thiswasrelatedtothevariabil- measured RNA concentration (Mathers et al. 1992). Although
ityinDNA.OtherresearchershavequestionedtheuseofRDas adownwardtrendinnucleicacidlevelsmayindicateaslowing
aproxyforgrowthinolderorlargerjuveniles(e.g.,Ciottietal. of growth rate, it does not necessarily reflect a decline in fish
2010) due to the poor observed correlation between RD and condition(Ch´ıcharoandCh´ıcharo2008).
growthinsomespecies(Ju¨rssetal.1987;DeBoecketal.1997; Our results affirm what other studies have also found: that
Kim and Kang 2004; Frommel and Clemmesen 2009). The RNA may be a more suitable condition index than RD among
RD-orRNA-basedgrowthestimatesderivedfromstudieswith juvenile fish of the same species (Chung et al. 1993; Wang
small fish may not be applicable to larger individuals, whose et al. 1993; Buckley et al. 1999), especially when DNA and
bodies contain less RNA-producing muscle tissue relative to fishsizearevariable.Sincetemperatureplaysanimportantrole
otherstructuralcomponents(FrommelandClemmesen2009). inregulatingmetabolism,proteinsynthesis,andRNA(Goolish
Inyoungfish,theproductionofproteinprimarilyfuelssomatic etal.1984),laboratorycalibrationexperimentsoverarangeof
NUCLEICACIDINDICESANDGROWTHOFJUVENILESCUP 5
RNA vs GWT 5.0
0.09 2 weeks RD Initial
4.5 Starved
4 weeks
0.08
6 weeks
4.0
0.07
3.5
WT D
0.06 R
G 3.0 * *
*
*
0.05
2.5
*
0.04 R = 0.757 2.0
P < 0.0001
N = 23
0.03 1.5
1.4 1.6 1.8 2.0 2.2 2.4 2.6 Initial 1 2 3 4 5 6 7 8 9
Mean RNA
DNA vs GWT 2.6
14 0.09 ht) 2.4 RNA
0 g
y 2 0.08 wei 2.2
Ma e 2.0
u
0 0.07 ss 1.8
eries] at 20:45 2 GWT 000...000456 R = 0.613 NA (ug/mg wet ti 1111....0246 * * * * *
h P < 0.0019 R 0.8
s
f Fi 0.03 N = 23 0.6
O 0.20 0.25 0.30 0.35 0.40 0.45 0.50 Initial 1 2 3 4 5 6 7 8 9
nt
e Mean DNA
m
rt
a DNA vs GLN
p 0.9
e 1.8
y [D ght) 0.8 DNA
d b 1.6 wei
oade 1.4 sue 0.7
Downl GLN 1.2 g wet tis 0.6
m 0.5
1.0 g/
u
A ( 0.4
0.8 R = -0.499 N
P < 0.0157 D
N = 23 0.3
0.6
Initial 1 2 3 4 5 6 7 8 9
0.20 0.25 0.30 0.35 0.40 0.45 0.50
Number of Days Starved
Mean DNA
FIGURE3. MedianRNA:DNAratio(RD),RNAconcentration(RNA),and
FIGURE2. ScatterplotsofmeanRNAconcentration(RNA)versusweight- DNAconcentration(DNA)forfastingScup(N=9–10fish)sampledeachday
based instantaneous growth rate (GWT); DNA concentration (DNA) versus overa9-dperiod.Asterisksindicateasignificantdifference(P<0.05)between
GWT;andDNAconcentrationversuslength-basedinstantaneousgrowthrate valuesmeasuredafter1–9doffastingandtheinitialvalue.
(GLN)ofjuvenileScupineachaquariumatthreeintervalsduringthefeeding
experiment.
6 MERCALDO-ALLENETAL.
temperaturesandfeedinglevelsshouldbeusedtogeneratenu- to 3 are known to indicate a starved condition in young fish,
cleicacid-basedgrowthmodelsforcomparingfishthatarecul- with critical values varying among species and with seawater
turedundermorevariabletemperatureconditionsorinthewild. temperature (Ferron and Leggett 1994; Meyer et al. 2012; De
Variationintemperature(<3◦C)wasslightduringour6-week Raedemaecker et al. 2012). The lowest RD value measured in
study and likely did not contribute to changes in biosynthetic fasting Scup (1.76) during our study is unlikely to represent
activity.TheuseofRNAasarelativeindicatorofgrowthorcon- theminimumthresholdforsurvival,assamplingendedbefore
ditioninScupshouldbelimitedtofishthathavebeencultured anystarvationmortalityoccurred.Weconcludethatnucleicacid
atsimilartemperatures. indicesmaybeusedformonitoringnutritionalconditionincul-
tured juvenile Scup since changes in biochemical constituents
NucleicAcid-BasedIndicesasConditionIndices were detected in unfed fish that were growing slowly or not
duringFasting atall.
TheRNAandRDinfood-deprivedfishdifferedsignificantly
from initial values within 5 d of fasting. Reductions in RNA Conclusions
and RD in juvenile fish have previously been reported (e.g.,
DecliningDNAconcentration,possiblyduetohypertrophy,
Bastrop et al. 1991; Chung et al. 1993; Mathers et al. 1993;
may explain the lack of a close relationship between RD and
Gwak and Tanaka 2001; Edwards 2009), with significant de-
growthrateinjuvenileScup.OurresultssuggestthatRDshould
creasesinRDorRNAoccurring2–8dafterfastingbegins(e.g.,
4 notbebroadlyappliedasaconditionindexforjuvenilefishwith-
1 BisbalandBengtson1995;Arndtetal.1996;SegninideBravo
0 outcarefulvalidationoftheRD–growthrelationshipforthelife
y 2 andChung1996;Rookeretal.1997;Pecketal.2003;Edwards stage, species, and size-class of interest. The concentration of
Ma 2009;Ciottietal.2010).Whenafishgoeswithoutfood,RNA RNA was strongly correlated with GWT in fed juvenile Scup
0 productionandproteinmetabolismaredirectedtowardmainte- andmaybeusefulforrelativecomparisonsofgrowthornutri-
2
5 nanceofcrucialbodyfunctionsratherthanbeinginvestedinto tionalconditioninfishofcomparablesizeandageundersimilar
4
0: growthprocesses(SegninideBravoandChung1996).BothRD culture conditions. The RD and RNA in Scup declined signif-
2
at andRNAinScuprespondedquicklytothewithholdingoffood, icantly within 5 d of fasting, while DNA remained relatively
s] asvaluesdecreasedby50%within9d.Otherstudieshavefound constant.Nucleicacidindicesmaybeusefulfordetectingfood
rie changes in RD to be subtle, to occur more slowly (Bisbal and limitation in juvenile Scup that are reared in an aquaculture
e
h Bengtson 1995; Rooker and Holt 1996; Weber et al. 2003), or
s setting. Our results confirm that the Scup is a potential candi-
Fi tobeundetectable(Richardetal.1991)inlargerfood-deprived
f date for marine aquaculture based on its ready adaptability to
O juveniles,whichhavegreaterenergyreservesthatallowthemto
pelleteddietsanditsrapidgrowthundercultureconditions.
nt withstand food deprivation without immediate changes in nu-
e
m cleicacidindices(Pecketal.2003;Weberetal.2003;Edwards
rt
a 2009).Generally,responsetimeofbiochemicalconstituentsto
p ACKNOWLEDGMENTS
e
D fooddeprivationvarieswiththespecies,lengthoffasting,fish
WethankElaineCaldaroneandRonaldGoldbergforeditorial
[
by size, and experimental conditions (Richard et al. 1991; Weber review;PaulClark,SarahPerry,andJamesReidyfortechnical
d etal.2003;Edwards2009). assistance;andCaptainsRobertAlixandWernerSchreinerfor
e
d The concentration of DNA did not differ from the initial
a fishcollection.Theuseoftradenamesdoesnotimplyendorse-
nlo pre-fasting concentration at any time over the 9-d starvation mentbytheNationalOceanicandAtmosphericAdministration,
w period. Many studies of fasting juvenile fish have found that
o NationalMarineFisheriesService.
D white muscle tissue DNA concentrations either remain stable
(Varnavskiyetal.1991;Arndtetal.1996;Kimetal.2008)or
increase(e.g.,Chungetal.1993,1998;GwakandTanaka2001;
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Use of Clove Oil and Eugenol to Anesthetize Fingerling
Shabut Barbus grypus
Fatih Öğretmen a , Selami Gölbaşi b , Burak E. Inanan c , Volkan Kizak d & Murathan Kayim d
a Faculty of Fisheries and Aquaculture , Muğla Sitki Kocman University , 48000 , Muğla ,
Turkey
b General Directorate of State Hydzraulic Works, Fish Production Station , 63300 , Urfa ,
Turkey
c Faculty of Science, Department of Biology , Muğla Sitki Kocman University , 48000 ,
Muğla , Turkey
d Fisheries Faculty, Tunceli University , 62000 , Tunceli , Turkey
Published online: 23 Jan 2014.
To cite this article: Fatih Öğretmen , Selami Gölbaşi , Burak E. Inanan , Volkan Kizak & Murathan Kayim (2014) Use of Clove
Oil and Eugenol to Anesthetize Fingerling Shabut Barbus grypus , North American Journal of Aquaculture, 76:1, 9-13, DOI:
10.1080/15222055.2013.824942
To link to this article: http://dx.doi.org/10.1080/15222055.2013.824942
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