Fish Protein Hydrolysates - FPH- enhance resistance of aquaculture species to different types of...

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International Aquafeed is published five times a year by Perendale Publishers Ltd of the United Kingdom. All data is published in good faith, based on information received, and while every care is taken to prevent inaccuracies, the publishers accept no liability for any errors or omissions or for the consequences of action taken on the basis of information published. ©Copyright 2012 Perendale Publishers Ltd. All rights reserved. No part of this publication may be reproduced in any form or by any means without prior permission of the copyright owner. Printed by Perendale Publishers Ltd. ISSN: 1464-0058 March | April 2012 Feature title: Fish Protein Hydrolysates - FPH- enhance resistance of aquaculture species to different types of stress The International magazine for the aquaculture feed industry

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Aquaculture production is now exceeding fisheries production in total seafood supply. Aquaculture, through intensification and husbandry practices, generate numerous stressful events, which may result from water deterioration (acute or chronic exposure to reduced levels of dissolved oxygen and/or increased levels of carbon dioxide, ammonia, nitrite) and/or handling (vaccination, transfer, transport, grading, ambient noise).

Transcript of Fish Protein Hydrolysates - FPH- enhance resistance of aquaculture species to different types of...

Page 1: Fish Protein Hydrolysates - FPH- enhance resistance of aquaculture species to different types of stress

International Aquafeed is published five times a year by Perendale Publishers Ltd of the United Kingdom.All data is published in good faith, based on information received, and while every care is taken to prevent inaccuracies, the publishers accept no liability for any errors or omissions or for the consequences of action taken on the basis of information published. ©Copyright 2012 Perendale Publishers Ltd. All rights reserved. No part of this publication may be reproduced in any form or by any means without prior permission of the copyright owner. Printed by Perendale Publishers Ltd. ISSN: 1464-0058

March | April 2012

Feature title: Fish Protein Hydrolysates - FPH- enhance resistance of aquaculture species to different types of stress

The International magazine for the aquaculture feed industry

Page 2: Fish Protein Hydrolysates - FPH- enhance resistance of aquaculture species to different types of stress

Aquaculture production is nowexceedingfisheriesproductionintotalseafoodsupply.Aquaculture,through intensification and hus-

bandrypractices,generatenumerousstress-ful events, which may result from waterdeterioration(acuteorchronicexposuretoreduced levels of dissolved oxygen and/orincreasedlevelsofcarbondioxide,ammonia,nitrite)and/orhandling(vaccination,transfer,transport,grading,ambientnoise).

Furthermore, environmental stressors,such as temperature and salinity variations,may also have detrimental consequences onaquaticcaptivepopulations.

Improvement of husbandry practices iscertainly the easiest way to reduce stressfuleventsandtheirconsequencesonzootechni-calperformance,thatisreducedgrowthrate,by sparing energy resources in homeostasisand increased mortality, by impairing innateimmuneandanti-oxidativestatus.

Beside husbandry practices, nutritional sup-plementationisanotherwaytofine-tuneimmuneandanti-oxidativedefensesofaquaticspecies.

In this context, nutrients like free aminoacids and derivatives, nucleotides, or anti-oxidants are often supplemented as immu-nostimulators,especiallyduringcriticalperiodssuch as weaning, wintering, seawater trans-fer.Whenproducedunderhighly controlledconditions, Protein Hydrolysates (PH) andespecially those of marine origin (FPH), arenaturallyrichinthesenutrients.

Furthermore, numerous studies demon-strated they were rich in bioactive peptidesofnutraceuticinterests(growthlikehormone,antioxidants, anti-stress and anti-microbialpeptides). The purpose of this article isto review the experienced benefits of PHon aquatic farmed species during differentinducedstressfulevents.

Variation of water temperature affects growth performances

Temperature and salinity variations are

the most experienced climatic issues. Whilecoldweatherusually impairs feed intakeandgrowth, it usually has less adverse effectsthanwarmweatheronaquaticspecieshealthstatus (oxidative stress, oxygen availability,resistancetoopportunisticpathogens).

A temperature challenge trial was imple-mented at the end of a feeding trial onEuropeanseabass(Dicentrarchuslabrax)juve-niles(meanweight:16.3g).

Fishwerefedwithisoproteic-supplement-eddiets(control,control+2.5percentFPH1and control plus five percent FPH1) for 29days in triplicate flow trough100ltanks.

At the end of the feedingtrial, 10 fish out 40 initial fishwerereplacedintheirrespec-tivetankforthethermalchal-lenge (duplicate). The range33-35°Chadpreviously beendetermined as lethal so tem-perature was increased from20°C to 33°C within twohours without any observedmorbidity (mortality plusunbalancedfish).

As shown in Figure 1observedmorbidityonlystart-ed at 34°C, after five hoursof challenge. The thermalchallenge was stopped twohours later when 80 percentof morbidity was observed.FPH1 supplementation didn’treduce final morbidity but itsignificantly delayed it (p-val-ue<0.01,proportionalhazardsmodels). This delay is espe-cially visible between 5.5 andseven hours, and it doesn’tseem to be dose-dependentfor 2.5 and five percent sup-plementations.

Without any metaboliteclues, we may only assumethat FPH1 has delayed the

physiologicalprocessresponsibleforfishmor-bidity, possibly thanks to hormone-like and/or to anti-stress effects. This trial may findcommercial applications for preventive feedsupplementationbefore the summer season,especially for flow-through or cage farmingsystems.

FPH enhance fish resistance to water quality deterioration

Through intensification, average waterqualityisoftenclosetothelimitstoleratedbyaquaticspeciesforanoptimalgrowth.

Fish Protein Hydrolysates FPH- enhance resistance of aquaculture species to different types of stress

Figure 1: European seabass morbidity during a thermal lethal challenge

Figures 2 & 3: Respective Nile tilapia and Tra Catfish survival rate during NH3 challenge trials (one way ANOVA/two way ANOVA)

by M Herault, V Fournier, M Hervy, A Ngoc, Aquativ–Za Du Gohélis 56250 Elven, France

12 | InternatIonal AquAFeed | March-april 2012

FEATURE

March-april 2012 | InternatIonal AquAFeed | 13

Based on this statement, any perturba-tion of the water quality, resulting fromoverfeeding, overcrowding, water renewalshortageoralgaeblooming,mayhavecriticalconsequencesonhealth&growthstatusesoffarmedspecies.

Twoammoniachallengetrialwere imple-mentedonTracatfish(Pangasiushypophthal-mus),andNiletilapia(Oreochromisniloticus),juveniles (mean weight = 13g) in order toassess the impact of FPHonoverall survivalrates.

Basically, fishwere fed for10weekswithcommercial diets, formulated with differentlevelsoffishmeal–FM-,supplementedornotwith FPH2 and FPH3 (1.8 percent in tilapiadiets & three percent in catfish diets). Afterthosefeedingperiods,10tilapiaand20catfishwere distinctly placed in aquaria (n=4 repli-cates)foranammoniachallengeconsistingina constant 150ppm TAN load for 48 hourswitha100percent/daywaterexchangerate.

Survival results are illustrated with Figure2andFigure3.

BothFPH2and3hadapositiveimpactonfish survival tohigh ammonia load challenge,this benefit being also observed on fishmealfree diet. As the end product of proteinmetabolism, ammonia is the most commontoxicant in culture and live-transportationsystems. It ismore toxicunder its unionizedformNH3 as it easily diffuses across the gillmembranereducingoutwardfluxofammoniaexcretion.

As a result, ammonia level in fish plas-ma increases causing various physiologicalresponses, in many ways similar to anoxiasymptomsandfinallyleadingtoneurotoxicityandfishdeathinmostseriouscases.

SeveralstudiesimplementedonFPHhavedemonstrated their biochemical functionali-ties,includingpotentanti-oxidativeproperties.

Thus, it becomes possible to increaseanimal’sownanti-oxidativedefensesthankstoFPHsupplementationand,asaconsequence,reduce expected adverse effects of exces-sive production of ROS -Reactive OxygenSpecies- resulting from anoxia like stressesinduced by many chemical and physical dis-turbances.

Restoring feed intake and growth during stressful events

It is a common practice to supplementaquaticfeedwithpalatabilityenhancerduringcriticalandstressfulperiodssuchasspawning,weaning,wintering,oralantibiotictreatments.

Improving growth ratesA Protein Hydrolysate was formulated

tomeetsuchneedsandserveasan internalreferenceforassessingnewlydevelopedFPH.A total of 28 short term feeding trials (13-21days)were implementedwith this internal

reference, coated at two percent dosageontoanutritionallybalancedPlantBasedFeed(PBF),within twodistinctexperimental facili-tiesnamed‘C’and‘D’.

These facilities, consisting in two serialsof 24 tanks of 100l capacity supplied withthermoregulatedandfiltratedseawater;onlydifferby their tankheights. Fish into facilities‘C’,duetolowertanks,aremoreexposedtotechnician’s work and visual contacts. Basedon the significant differences observed foraverage zootechnical performance indicators(see Figure 4), this visual disturbance stressobviouslyimpactsfishbehavior.

Thus,growthof fishreared in facilities ‘C’isonlyat67percentoftheaveragepotentialgrowthobservedforfacilities‘D’.Thisgrowthretardation is mostly explained by a muchlower feed intake, which may entirely besolvedbyPHsupplementation.

In addition, through PH supplementation,growth rate is finally improvedbyalmost20percent,duetoacombinedimprovementoftheFCR,by10percentonaverage.

Thisbenefitcombinationresults,onaver-age, in a 28 percent growth rate improve-ment under normal conditions and almost80percentgrowthrateincreaseundervisualdisturbanceconditions.

Atthismacroscopicscale,mostlikelyassump-tionsmaybethefollowings:FCRimprovementprobably reflects areduction of PBFinducedgutinflam-mation and/or anoptimization ofgut florawhile thehigher feed intakerecorded with PHsupplementationis certainly due tofeedenhancedpal-atability.

FPH improve fish resistance to opportunistic pathogens

Chronic stress,because of con-tinuoussecretionofcortisol, is harmfulasitleadstoimmu-nosuppression andthereby increasesrisksofdiseaseout-breaks.

A disease chal-lenge was initi-ated on juvenileNile tilapia (meanweight: 37.7g±2.2)followinga42days

feedingtrialimplementedwithcontrolcommer-cialfeed,withorwithoutFPHsupplementation(twopercentdosage).SeeFigure4.

1.0ml of a bacterial suspension consistingin9x108UFCofAeromonashydrophilawasinjectedintothefishabdominalcavity.Therewerefourreplicatesand15fishperreplicatewere inoculated and then monitored forAeromonashydrophilasymptomsfor10days.

Attheendofthisperiod,averagesurvivalrate of inoculated fish for control feed wasveryhigh,whichmeansthat initial fishhealthstatus was really good before starting thediseasechallengetrial.

Despite this high survival rate, FPH sup-plementation significantly improved overallsurvival rates from 88 up to 98 percent(p<0.02).At thismacroscopic scale, it isdif-ficult to discuss how FPH has improved fishresistancetoopportunisticpathogens.

Wemay,however,proposetwodifferentandmaybecombined,patterns.

First, FPH, thanks to its bioactive func-tionalities (anti-stress and immunostimulantmostly),mayhaveimprovedoverallfishhealthstatusbeforeand/orduringthechallengetrial.

Second,FPHmayhavebeenanaturalsourceof Anti Microbial Peptides, which may havebeen effective against Aeromonas hydrophila.Biochemicalstudieswouldbenecessarytocon-firmone,orboth,oftheseassumptions.

12 | InternatIonal AquAFeed | March-april 2012 March-april 2012 | InternatIonal AquAFeed | 13

FEATURE

Based on this statement, any perturba-tion of the water quality, resulting fromoverfeeding, overcrowding, water renewalshortageoralgaeblooming,mayhavecriticalconsequencesonhealth&growthstatusesoffarmedspecies.

Twoammoniachallengetrialwere imple-mentedonTracatfish(Pangasiushypophthal-mus),andNiletilapia(Oreochromisniloticus),juveniles (mean weight = 13g) in order toassess the impact of FPHonoverall survivalrates.

Basically, fishwere fed for10weekswithcommercial diets, formulated with differentlevelsoffishmeal–FM-,supplementedornotwith FPH2 and FPH3 (1.8 percent in tilapiadiets & three percent in catfish diets). Afterthosefeedingperiods,10tilapiaand20catfishwere distinctly placed in aquaria (n=4 repli-cates)foranammoniachallengeconsistingina constant 150ppm TAN load for 48 hourswitha100percent/daywaterexchangerate.

Survival results are illustrated with Figure2andFigure3.

BothFPH2and3hadapositiveimpactonfish survival tohigh ammonia load challenge,this benefit being also observed on fishmealfree diet. As the end product of proteinmetabolism, ammonia is the most commontoxicant in culture and live-transportationsystems. It ismore toxicunder its unionizedformNH3 as it easily diffuses across the gillmembranereducingoutwardfluxofammoniaexcretion.

As a result, ammonia level in fish plas-ma increases causing various physiologicalresponses, in many ways similar to anoxiasymptomsandfinallyleadingtoneurotoxicityandfishdeathinmostseriouscases.

SeveralstudiesimplementedonFPHhavedemonstrated their biochemical functionali-ties,includingpotentanti-oxidativeproperties.

Thus, it becomes possible to increaseanimal’sownanti-oxidativedefensesthankstoFPHsupplementationand,asaconsequence,reduce expected adverse effects of exces-sive production of ROS -Reactive OxygenSpecies- resulting from anoxia like stressesinduced by many chemical and physical dis-turbances.

Restoring feed intake and growth during stressful events

It is a common practice to supplementaquaticfeedwithpalatabilityenhancerduringcriticalandstressfulperiodssuchasspawning,weaning,wintering,oralantibiotictreatments.

Improving growth ratesA Protein Hydrolysate was formulated

tomeetsuchneedsandserveasan internalreferenceforassessingnewlydevelopedFPH.A total of 28 short term feeding trials (13-21days)were implementedwith this internal

reference, coated at two percent dosageontoanutritionallybalancedPlantBasedFeed(PBF),within twodistinctexperimental facili-tiesnamed‘C’and‘D’.

These facilities, consisting in two serialsof 24 tanks of 100l capacity supplied withthermoregulatedandfiltratedseawater;onlydifferby their tankheights. Fish into facilities‘C’,duetolowertanks,aremoreexposedtotechnician’s work and visual contacts. Basedon the significant differences observed foraverage zootechnical performance indicators(see Figure 4), this visual disturbance stressobviouslyimpactsfishbehavior.

Thus,growthof fishreared in facilities ‘C’isonlyat67percentoftheaveragepotentialgrowthobservedforfacilities‘D’.Thisgrowthretardation is mostly explained by a muchlower feed intake, which may entirely besolvedbyPHsupplementation.

In addition, through PH supplementation,growth rate is finally improvedbyalmost20percent,duetoacombinedimprovementoftheFCR,by10percentonaverage.

Thisbenefitcombinationresults,onaver-age, in a 28 percent growth rate improve-ment under normal conditions and almost80percentgrowthrateincreaseundervisualdisturbanceconditions.

Atthismacroscopicscale,mostlikelyassump-tionsmaybethefollowings:FCRimprovementprobably reflects areduction of PBFinducedgutinflam-mation and/or anoptimization ofgut florawhile thehigher feed intakerecorded with PHsupplementationis certainly due tofeedenhancedpal-atability.

FPH improve fish resistance to opportunistic pathogens

Chronic stress,because of con-tinuoussecretionofcortisol, is harmfulasitleadstoimmu-nosuppression andthereby increasesrisksofdiseaseout-breaks.

A disease chal-lenge was initi-ated on juvenileNile tilapia (meanweight: 37.7g±2.2)followinga42days

feedingtrialimplementedwithcontrolcommer-cialfeed,withorwithoutFPHsupplementation(twopercentdosage).SeeFigure4.

1.0ml of a bacterial suspension consistingin9x108UFCofAeromonashydrophilawasinjectedintothefishabdominalcavity.Therewerefourreplicatesand15fishperreplicatewere inoculated and then monitored forAeromonashydrophilasymptomsfor10days.

Attheendofthisperiod,averagesurvivalrate of inoculated fish for control feed wasveryhigh,whichmeansthat initial fishhealthstatus was really good before starting thediseasechallengetrial.

Despite this high survival rate, FPH sup-plementation significantly improved overallsurvival rates from 88 up to 98 percent(p<0.02).At thismacroscopic scale, it isdif-ficult to discuss how FPH has improved fishresistancetoopportunisticpathogens.

Wemay,however,proposetwodifferentandmaybecombined,patterns.

First, FPH, thanks to its bioactive func-tionalities (anti-stress and immunostimulantmostly),mayhaveimprovedoverallfishhealthstatusbeforeand/orduringthechallengetrial.

Second,FPHmayhavebeenanaturalsourceof Anti Microbial Peptides, which may havebeen effective against Aeromonas hydrophila.Biochemicalstudieswouldbenecessarytocon-firmone,orboth,oftheseassumptions.

12 | InternatIonal AquAFeed | March-april 2012 March-april 2012 | InternatIonal AquAFeed | 13

FEATURE

Page 3: Fish Protein Hydrolysates - FPH- enhance resistance of aquaculture species to different types of stress

SA B E T H A , K S —Februar y 2012 —TheWe n g e r E x t r u s i o n

Group (comprised ofWengerManufacturing, Inc, Extru-Tech,Inc , and Corporate ProjectSe r v i ce s ) ha s announcedthe acqu i s i t ion o f SourceTechnology A/S, the leadingprov ider of in l ine ana lys i ssystemsusedinfoodandfeedmanufacturing. Headquar teredin Kolding, Denmark, SourceTechnology suppl ies in l inesampling and analysis systemstofourkeyindustriesinvolvingpelletsandpowders.

Source Technology lever-ages its innovative exper tisetohelpmanufacturersenhanceproductquality,improveenergyand operational effic ienciesand increase foodsafetyregu-

lationcompliance.Typicalappli-cations for SourceTechnologyinline analysis devices includemeasur ing of bu lk dens i ty,tap density, moisture content,product sizing, par ticle sizing,NIR analysis, burned par ticled e t e c t i o n , c o l o r i n t e n -s i ty, dust test ing , durabi l i tytesting, floating test and muchmore.

T h e We n g e r E x t r u s i o nGroup , headqua r te red i nSabetha, Kansas, is a globaldesigner,manufacturerandfullser vice provider of extrusionprocessing systems.Theacqui-sit ion of SourceTechnologywill strengthen their ability toprovide a full scope of auto-mated extrusion technologysystemswithenhancedprocesscontrol.Wenger systems are

installed in over 90 countries,and are suppor ted through330 extrusion-specific profes-sionals based in twelve globallocations. Wenger manufac-tures and suppor ts the indus-t r y ’s most comprehens iveser ies of extruder s , dr yer sand controls for the commer-cial production of pet food,human food, aquatic and live-stockfeeds.

More inforMation:Lafe Bailey, Wenger Vice President of Sales and ServicesEmail:[email protected]

Thomas Jorgensen, Source Technology PresidentEmail:[email protected]

Website:www.wenger.comWebsite:www.sourcetechnology.dkWebsite:www.extru-techinc.com

WengerExtrusionGroupannouncespurchaseofSourceTechnologyA/SInlinesamplingandrealtimeanalysisbecomeskeytofullintegration

6 | InternatIonal AquAFeed | March-april 2012 March-april 2012 | InternatIonal AquAFeed | 7

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The overarching objectiveshould be to promote aquacul-ture as a viable wealth creating,

undertaking, and laying emphasis on pri-vate sector-led initiatives with govern-mentsprovidingtheenablingenvironmentfor growth. The increased growth couldbe realised through improvements intechnologies and resource use, 41 inte-grationofaquaculturewithother farmingactivities accompanied with appropriatepolicies and strategies in marketing andtrade.

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• Developing strategies for value addi-tion for aquatic products such as bydemonstrating high quality control,improvebiosecurityandaquaticanimalhealthmanagementandbyunder takingmarket/product-relatedresearch ■

28 | InternatIonal AquAFeed | March-april 2012 March-april 2012 | InternatIonal AquAFeed | 29

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Page 4: Fish Protein Hydrolysates - FPH- enhance resistance of aquaculture species to different types of stress

Meanwhile,thisassayconfirmsthisispossi-bletoenhancefishresistancetoopportunisticdiseasebyFPHsupplementation.

ConclusionsDespite of continuously improved aqua-

culture practices, stressful events will remaininherenttohusbandrybothbecauseofclimat-ichazards, fornonRecirculatingAquacultureSystems, and human operations. Stress is anormal and adaptive process (homeostasis)consistinginthreechronologicalresponses:

1) primary responsesinvolve the elevation ofplasma catecholaminesandcortisolhormone;

2) secondary responsesrefer to the effects ofthese hormones attissue level; 3) tertiaryresponsesrefertoanimalzootechnical perform-ancessuchaschangesingrowthandhealth.

Minor punc-tual stress willmostly be lim-ited to primaryand second-ary responseswhile acuteand chronicstress will ofteninduce the terti-ary response’sadverse effects.As previ-ously illustrated,Fish ProteinHydrolysatesare the idealnutritional solu-tiontoattenuatestressful event’sadverse effectson zootechnicalperformances.

Thanks totheir naturalhighcontentsoflow molecularweightNitrogencompounds,which includenucleotides,aminoacidsandderivatives, andbioactive pep-

tides, FPH will act directly, or indirectly, onaquatic species feed acceptance, feed trans-formation, innate anti-oxidative and immunedefenses,whichwillendupwithrestored,orenhanced,growthrateandhealthstatus.

Beside their high nutritional value, FishProtein Hydrolysates are therefore a goodalternativetopreventiveandgrowthpromot-ing,antibiotictreatments,especiallywiththeiranti-microbial properties and to any otherfeed additives used as immune-stimulants oranti-oxidativeagents.

Last but not least, the bioactive peptidesprofile,thereforethephysiologicalresponse,isdrivenbyhydrolysisprocessleveragessuchastypeofenzyme,temperaturextimescheduleandpH. ■

Referencesavailableonrequest

Figure 4: Zootechnical performances observed on E. seabass juveniles (initial mean weight: 4.7-11.1g) over 28 trials implemented within two facilities “C” and “D”

Figure 5: Survival rate of Nile tilapia to an Aeromonas hydrophila challenge, 10 days following injection (one way ANOVA)

for More inforMation:AquativZ.A. de GohelisElven, 56250 FranceTel:+33297938080Email:[email protected]:www.aquativ-diana.com

14 | InternatIonal AquAFeed | March-april 2012

FEATURE

March-april 2012 | InternatIonal AquAFeed | 15

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Page 5: Fish Protein Hydrolysates - FPH- enhance resistance of aquaculture species to different types of stress

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Filtration in recirculation particle control

Fish Protein Hydrolysates– FPH enhance resistance of aquaculture species to

different types of stress

The quest to keep Salmon in the pink naturally

Aquaculture development in Sub-Saharan Africa

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