GUAM-T-96-001 C2

Final Report

Pacific Aquaculture Association

Culture of Tropical Marine Aquarium Fishes By: Jeffrey A. Tellock

TECHNICAL REPORT NO. 020

JUNE I 996

TECHNICAL REPORT SERIES OF THE GUAM AQUACULTUREDEVELOPMENT AND TINNING CENTER

Departmentof Commerce Government of Guam 102 M. Street Tiyan, Guam 96913

TABLE OP CONTK'ATS

SUlvCvbQtY OF FINDINGS

PROJECT OBJECTI'&S

METHODS

RESULTS

DISCUSSION OF RESULTS

RECOMMENDATIONS FOR CULTURE OF hhVW< AQUARIUM SPECIES

CONCLUSIONS 50

LITERATLtRE CITED

PERSONAI. COhQAK'4ICATIONS 53

APPENDIXES

I FROZEN DIET FOR hGQUNE AQUPDGL'M FISH 54

II AQUILAi;4!ej HOBBY~RATUItE ON THE CULTURE OF 14bQt24EAQUPduUM FISH AND SHMhP

III SELECTEDSCIENTIFIC LITERATURE ON THE CULTS 62 OF MAPS& FISH AND SHIt2vP

IV SELECTED SCIENTIFIC LITERATLIRE ON SPA%~G 68 AND LARVAL BIOLOGY OF MAIUNE AQUPdUUMFISH AND SHI598P

V LIST OFMPdUNK AQUARIUM FISEES REARED IN 70 C/ZTPv'ITY

VI LIST OF MPdUNEAQU/ELM SHIulVP REAREDIN 72 CPS'/ITY ACKNOW'%.EDGMENTS; TheGADTC staff enjoyed conducting the project and would like to thankthe Pacific AquacultureAssociation forthe opportunity to do so. I wouldspeci6cally liketo thank theprogram director. Anne Bailey, who provided contmuous support and Rich Bailey for reviewingthe project.! woukl like to thankthe following GADTC stafF members fortheir assistancewith this project: Carl KitteL Wmg-Kai Wang, Victor Camacho. Frank Alig, MarkNorman, Daphane Peralta and Dana Edwards. Former GADTC staff members DanteBueasuceso, Leona Figir3g and Pedro Salas also assisted with the project.Ia additioa,I would like to thankthe Director of theDepartment of Commerce, Frank B. AguonJr., for his support and involvement. Wewould especially bketo thankthe Chief Plannerfor theEconomic Development and Plaanmg division of theDepartment of Commerce,Wilbam FitzGerald, and his successorsLinda Flynn and IaterRichard Carandangwhoall provided continuous supervisioa andadvice. I would also like to thank ClydeTamaru of HawaiiC's Aquaculture Consultant Services for assistingme in cannulatingclownfish. spawnmg sea urchms, aad taking photographs of developmental stagesoflarvae. Iaaddition. heprovided advice on several aspects ofthe project. ! would hketo especiallythank Arnie Moreao for geaemusly donating fish for use as broodstock, andfor assisting mein collecting oysters for the project. Ian Elhot and Richard Sakamoto alsoprovided fish for this project. I would like to thankGerry Davis for information on regulatoryissues conceraiag marine tropical fish and invertebrate culture. Robert Myers andRichard Pyle also provided information oathe marine aqmlrhnn trade. Finally,! would liketo thankthe following people for providinginform«tion ontheir involvement ia researchor commercial production of marine tropical 5sh «nd shrimp: William Addison, HarryAko. Chris Brown, Stanley Brown, Leroy Creswell, David Fletcher, Tom Frakes. JoanHo1t, Frank HofF. Syd Kraul, Joe Lichteabert, Martin Moe, Linda Nelson, Godfrey Waugh,and Forrest Young. PACIFIC AQUACI. LTURK ASSOCIATION

PROJECT RKPORT

FINAL REPORT

PROJECT TITLE Culture of Tropical Marine AquarnnnFishes

PROJECT PERIOD September 1994 to March 1996

PRINCIPAL INVESTIGATOR Jeff Tellock Biologist-IV GuamAquaculture Development and Training Center EconomicDevelopment and Planning Division Guam Departmentof Commerce 102 M Street Tiyan, Guam 96913

SU7iQNARY OF FINDINGS

Many new speciesof marinetropical 6sh and shrimp have been cultured m recent years and some of these are beginning to be produced at a commercial level. However. clownfishare still the main product of marineaquarian 6sh hatcheries.Commercial efforts are becomingmore successfiilRom a biological standpoint,but someare stiH struggling 6nanciallyto make this type of aquaculturepro6table. Some commercialmarine tropical fish hatcherieshave discontinuedoperations after severalyears of activity, while others havestarted up m recentyears or may start up in the near batture.

Researchactivity in this area is also occurring at the university level, l.iterature on the culture of marinetropical Qshand shrimpis increasing,but occursmamly in the aquarium hobbyhterature. rather thanthe scienti6cliterature,

A list of key marmeaquarium fish and shrimp available in Micronesiaand conducive for tnariculturewas compiled,and someof these speciesare particularlypromising for commercial culture. Culture methods were eiuLmined for the Cinnamon clownfish .4. HNAE. REPORT - CULTLHK OF hbVUNE AQUPdULM FISHES 92 rnelanopus.the gobiesPtereleotris -ebra and Valencienneastngatvs. and the Banded coralshrimp Stenopus hispidus. Several batches of A. melanopuswere successAdiy reared to the juveaile stage, Regular spawaiagwas obtainedfor the gaby broodstock. but no 1arvaewere reared beyond the first-feedingstage for eitherof the gobyspecies. Larvae of S. hispiduswere reared through the earlylarval stages, but noaesurvived the long larval period to becomejuveniles.

Problemsencountered in the culrure of clowafishwere mainly ia the production of eggs. Success|ullarval culture of thegaby larvae was not achieveddue to a lackof a suitable firstfood for these tiny larvae. Progress was made in thelarval culture of theBanded coral shrimp,but eventualcontamination of larvaltanks by hydroidsaad jellyfish medusae resultedin completemortality before the larvae reached the juveaile stage. Suggestions are presentedto overcomeall of theseobstacles in thereariag of theseand other species of marinetropical fish and shrimp.

A growing tread of interestto commercialaquarian fish producersis the increaseof restrictionsoa the importationaad exportatioaof marinetropical species,with some islandnations closing down or severelyrestkctiug exports of reef fishesand invertebrates. Guamis preparmgto prohibitthe exportof marinefish aad invertebrates fiom its reefs. Thismay be a growmgtrend among island nations as they seek to protecttheir resources. Somecountries are also begmamgto restrict imports of meme tropical fish and invertebrates. Aneconomic analysis of marinee~ariutn fish culture suggests that commercial culture of variousmarine ornamental species could be pro8table in Micronesia.The most inrportant factorsaffectin profitabihtyare consistent production of marketable6sh, market price aadsales volume. Feed costs are aot a majorfactor a5~ng economicreturn, although the unit costof feedfor marmeornamental Ssh is machhigher than commercialdiets for foodfish.Economic projectioas were made for a smallhatchery aad growwut facility raisingclownthh.

PROJECT OBJKCTIVKS

Theproject wasdesigned to meetthe following objectives: I. Ideatifycur@mt private aad government efforts to culturetropical marme aquarium fishesthroughout the world. 2. Conductapplicable literature review for theculture of marinetropical fishes.

3. Identificatioaand listing of keymarine aquarium fishes readily available on Micronesian reefsaad lagoons that are coaducive for maricultureapplication.

4. Choosetarget fish species and explore culture methods.

PREPARED BY GADTC FINAI REPORT - CULTURE OF hbVUNK AQUAIUUM FISHES

5. Identify currem aad anticipatediateraatioaal regulatory coastrunts to the import!export of variousmarine aquarium 6shes.

6. Determmemethods of mariculturecertification of restrictedspecies.

7, Identify!determineeconomic parameters that would makethis new form of mariculture feasibleon a cotaatercialscale; from culture/growout to export.

METHODS:

Objective 1. Private and goverm2nentefforts to culture tropical marme aquarium fishes were identified through literature review and telephonecontacts. In addition, database reports were obtainedfrom the Breeder'sRegistry, a newsletterand databaseproviding informationoa breofmgaad propagationof marineaquarium 6sh aad invertebrates.A video from a conferenceon maxinetropical 6sh culture aad another video about a small comme' mtu~ aquarium6sh hatchery also provided information oa previous efforts at propagatingmzIme tropical 6sh.

Objective 2. A review of the literature was conductedfor the culture of marme tropical aquaritunfish and invertebratesthroughout the study. A broader review of the literature oa culture methodsfor other marine6sh was conductedin the later stagesof the study,

Objective 3. A list of key marine aquarium 6shesavailable in Micmnesia and conducive for maricuitmeapplicatMm was created,using the foIowiag criteria: availabihty,popularity in the aquluitnn trade, wholesaleor retail vahe, known adult and larval characteristics e.g., size of spavttners,spawn size, egg and larval size, larval duration!, available mformationon |ynvmag and larval rearing, reports of successfuladture efforts, and ease of culture of relatedspecies. Not all the speciesin the list quaii6edfavorably under every criteria.but eachspecies qualified under at leastsome of the criteriaand show potential for eventual commerce culture. Scietxti6c and common aames were checked and veri6ed through two guidebooks on mluiae aquarium6shes t Burgesset al., 1990;Myers. 1991!.

Objective4. The following marine aquariumspecies were obtaiaedfrom local collectors for possibleuse in this project:

Species Commoa name Number Amphipnorrchrysopterus Blue-stripeclowa6sh 3 pairs Arrtphiprionclarkii Clark's clowafish 2 pcs Amphiprion melarwpus Cianamon clowafish 30 6sh Amphiprionpenderaion Pmk skunk clowafish 1 pair Ptereleotris zebra Zebra goby 1 pair Valencienneastrigatus Blue-streak goby 1 pair Stenopushispidus ' Bandedcoral shrimp 8 pairs

PREPARED BY GADTC FINAL REPORT - CULTURE OF Mc~E AQUPdUV4} FISHES

' Although this projectoriginaHy targeted aquarium 6sh oaly, it was later expandedto includethe Bandedcoral shrimp.

The majority of the fish and three pairs of the Bandedcoral shrimpwere collectedduring the first six monthsof theproject. The Cmnamoa clownfish, A. melanous,were coHected as three groupsof fish. Initial effortswere madeto pair up the clowafishby putting together one large and one smaHerfish Severa} iadividua}swere also canaulatedto idetttdymale and female6sh anddetermine the level of taaturation.Another group of Cinnamonclownfish coHectedlater in the study was placed togetherwith severalother unpaaedclownish in a large0-ton! fiberglasstank coatailiingseveral spawning sites i.e., bricks and tiles! and anemones.The other speciesof clownish and gobieswere coHectedas matedpairs. These6sh were aot canaulatedto determinethe level of maturation,so as aot to risk damagingtheir reproductivetracts.

Various sizesof 6berglasstanks at the GuamAquaculture Development and Training Center GADTC! were used for holding cloem6shbroodstock, ranging horn about 400 liters to 2000 hters. Some of the fish were hek} inside the hatchery and others were hek} outdoors.Ceramic tiles were placed in thetanks containing c}os~ fot'use as spawnmg substrates.The tiles were exammeddaily for eggs,usually in the morning.Pairs of Banded coralshrimp were also placed in threeof the broodstockholding tanks. Later in the study, five additionalpairs wete collectedand aH the Bandedcoral shrimp were moved to two shaHowculture trays inside the hatchery,with eachpair placedm a separatecompartment Figure1!, Waterquality was maintained m aHthe broodstock tanks and grow out tanks by a continuousflow of seawaterthrough the tanks.The seawaterat the GADTCis obtained Rom saltwater weHs, without aay additional treatment or fi}tration. Indoor broodstocktanks received natural sunhght which came into thehatchery.

Thepair of Blue-streakgobies, V. strigutus, were miuntamed in a 3000liter tank insidethe hatchery.The tank was provided with a shahowtray fiHed with coral sand and short pieces of PVC pipe stuckinto the sandfor a spavmmgsite Figure2!. ThePVC pipewas checkedoa a regularbasis for the pzesenceof eggs.The behavior of the fishwas also observedfor indicationsof splvaiag. Thepair of Zebragobies, P. zebra,were maintained outdoorsin a 1000hter tank. Giant clams sheHs were provided on thebottom of the tank. formingsmall caves that the fish could go insideand usefor a spawningsite. The clam sheHwas checkec}oa a dailybasis for the presenceof eggs.

Thebroodstock were primarily fed a &ozendiet prepared at theGADTC. The recipe for this diet was similarto a gelatindiet usedat the WaikikiAquarium, with some modificatioas.Ingrediettts used m thediet and iastre~ons for prepariagit are listed in Appendix1. Thecomposition of the diet varied slightly 6'om batch to batch,depending on localavailability of ingredients.The broodstock were usually fed ad libitumtwice daily with the 6'ozenfood. Ia the beginnmgof the study,this diet was occasionally supplementedwith liveadult Anemia or sma}1penaeid postlarvae. Later in thestudy. brine

PREPARED BY GADTC Figuel. Bandedcoral shrimp broodstock tanks. It wasdetermined thatthese 20 and 30 litertanks were adequate to maintainspawning pairs of Bandedcoral shrimp.

Figure2. Blue-streakgoby broodstock and pvc pipe spawning site.

PREPARED BY GAQTC FINAL REPORT- CULTUREOF MARINE AQUA$QL'MFISHES ~ ~ shrimpflakes were occasionally usedas a supplementalfeed.The banded coral shrimp werealso fed a shrimpmaturation pellet on a regularbasis in addition tothe frozen food. Clownfishegg development wasmonitored visually and by length of incubationperiod. Previousexperience andreports in the literature indicated that the egg incubation period ofclownfish isusually between sixand nine days, depending ontemperature andcondition of thebroodstock. Late in theafternoon on the day when a batchof eggsappeared to be fullydeveloped andready to hatch,the tile on which the eggs were laid was removed Bom thebroodstock tank and put into a larvaltank. The tile was placed on edge and leaning againsttheinner standpipe, withan airstone placed near the egg mass, sothat a streamof airbubbles passed close to the egg mass. The following day after the eggs hatched, theti1e wasremoved and returned to thebroodstock tank. A few spawnswere deposited on the sideof the broodstocktank or hatchedearher than expected.In thesecases. it was necessaryto remove the larvae Qom the broodstock tank after hatching. Newly-hatched ciownfishlarvae are positively phototactic and were concentrated with a lightand siphonedmto a bucketof seawater. Bandedcoral shrimp egg development was monitored regularly based on the appearance ofthe egg mass. New egg masses area blue-greencolorand become a creamy white as theydevelop. When the eggs are ready to hatch, dark eyespots canbe seen in the white eggs.A femalewith mature eggs was placed into a bucketof clean seawater with an airstonein the afternoon preceding hatching. The next mornmg, the female was returned toits holding tank and larvae were counted and stocked into a larvalrearing tank. Spawnsize in clownish was estimated bycountmg the"egg spots" on the spawning site akerhatching. These egg spots usually appeared aslighter colored spots on the tile where theeggs were attached. Spawn size was estimated in Banded coral shrimp by counting thenumber of larvaein several6-ml samples taken from a bucketcontaining the hatched larvae,calculating anaverage number of1arvae perml, and multiplying thatnumber bythe total volume of the bucket. Threetypes oflive foods were examined inlarval rearing trials, inchding rotifers both s- typeand ss-type!, seaurchin larvae and oyster trocophores. Rotifers were mass cultured outdoorsin batchculture as described byTamaru et al.993!. TheGADTC originally culturedthe s-strain of rotifer Brachionus plicarrlis!, but m April1995 production was switchedover to the ss-strainof rotifer 8. rotuIrdus!. Seaurchins Zchinometra mathaei! were collected f'rom local waters and maintained in outdoortanks, where they grazed upon the algae growing on the sides of thetanks. Fertilizedsea urchin eggs were obtained by inducedspawning after injecting approximately0.5ml of a 0,5MKG solution into the body cavity ofthe sea urchins using a 1cc tuberculin syringe, The injected individual was then placed upside down in a 250ml glassbeaker filled with seawater andobserved forthe release of gametes. Males and feinaleswere identified bythe release ofsperm or eggs, respectively. Males were removed

PREPARED BY GADTC FINAL REPORT- CULTURE OF MAIUNE AQUARIUM FISHES ~ ~ petridish so that milt could be collected without mixing with sea water. Females were allowedtospawn until eggs no longer were released. Thespawned eggs were collectively placedinto a smallbeaker containing seawater. The sperm was activated with seawater andthen a fewml of dilutedspermic was added into the container of spawnedeggs and mixedto fertilizethe eggs. The eggs were then sieved through a 150micron screen to removelarge debris and placed into a 3-literpitcher of seawaterand aerated until used. Tamaruand Carlstrom-Trick 995! provideadditional details of thespawning procedure anddevelopmental sequence o fthe sea urchin larvae used in thisstudy. Oysters Saccosrrea cucullata tuberculata and/or Crassostrea echinata! were collected in SasaBay, Guam and held in tanks at the GADTC. The adult oysters were occasionally fed algaecultured atthe GADTC. Some ofthe oysters were held in tanks containing juvenile mulletor tilapia.Oyster trochophores were obtained after natural spawning probably followinghandling stress! or byplacmg them in anaqua'irium andadding approximately fourppm hydrogen peroxide Figures 3 and 4!. Oyster trocophores were harvested with a 23 micronscreen about 10 to 12hours after spawning and added to the larvaltank.

Clownfishlarvae were reared m 500liter blackfiberglass tanks. Larval tanks were initially filled to about250 to 300 htersvolume with seawaterand provided with Bghtaeration. Severalliters of a denseculture of microalgae Chaetoceros gracilis or A'annochloropsis oculata!was added to thelarval tanks daily during the first two weeks of thelarval period to providefood for the rotifers and to helpmaintain water quality. The amount of algae added usually 5'fo to 10'/oof thetank volume! depended on thedensity of thealgae culture.the amount of algaealready present in thelarval tank, and the volume of thelarval tank.Artemia nauplii was added to the larval tanks, beginning about seven days posthatch. Artificialdiets i.e., ground up artemia flake, larval fish diets, or frozen food! were usually addedbeginning at about day 12 to 15posthatch. Daily water exchanges were initiated afterthe first week, and a continuousflow of waterwas started after the first twoweeks to maintainwater quality. Juvenileclo~rnfish were grown out in fiberglasstanks of 1000to 3000liters volume both indoorsand outside. Juvemles were fed twice daily with the frozendiet preparedat the hatchery.Occasionally, thisdiet was supplemented withbrine shrimp flakes, Fish were occasionallyhand-sorted bysize and marketable fishselected forsale to a localretail pet store or to a commercial fish collector for export. Severallarval rearing trials were conducted with the Bandedcoral shrimp. In the beginningofmost of the trials. rotifers were maintained inthe larval tank at a densityof10 to20 rotiferslml and the green algae Nannochloropsis oculata was added daily to maintain a concentration.of about 500,000 to 1,000,000cells/ml in thelarval tank. After the first twoto threeweeks. the diet was gradually changed from rotifers to Anemianauplii and eventuallyenriched 24-hour old Anemia. Daily water exchanges were begun after about thefirst week, and a slowcontinuous flow of seawaterwas introduced to the larval tank after about three weeks.

PREPARED BY GADTC F1NAL REPOR - Ci 'LTURE OF MAR1%iEAQU XNL>il F1SHFS

~i~ure ~. Oyster spawning tank before the release of gametes.

Figure: 4. . ys0 t,pawnin~~ er spa~ tankafter the release of garnet~s into the water. This spawn producedviable oyster larvae.

PREPARLD BY GADTC FINAL REPORT - CULTL'ItE OF M/dtINE AQUARIUM FISHES

Objective 5: Regulatoryconstraints were investigatedby personalcommutucations with Gerry Davis, FisheriesSupervisor at the Division of Aquatic and Wildlife Resources DAWR!, Guam Departmentof Agriculture. Robert Meyer formerly with the DAWR!, and RichardPyle Bishop Museum.Hono1ulu. HI!,

Objective 6: Methods of mariculture certification were investigatedby personal communicationswith GerryDavis of the DAWRand by reviewof the literatureand other maricultureoperations.

Objective7: Cashflow projections were made for a smallclownBsh hatchery and grow- out facility,

RESULTS:

Objectivel. A few hatcheriesare known to be currentlyproducing marine aquarium fish commercially.The largesthatchery is C-Quest,Inc. in PeurtoRico. Anotheris Reef Propagations,a smallbasetjneat hatchery in Chicago.A third companyis DesertFisheries in Utah. A fourthhatchery believed to be the Tropica1Marine Centre Ltd.! is 1ocatedin England.'Hzee other hatcheries are presently operating at a researchand development or pilot stageand are supplymg small numl:mrs of tank-raised6sh to the marmeaquarium market.These include a hatcheryowned by RedSea Fish pHarm, Ltd. in Israel andtwo closedsystem hatcheries in the UmtedStates owned by SeaPhiz.Four commercial hatcheriesformerly produced maim' aquatiiim 5sh, but are no longerin operation.These includeAqualife Research Corporation in theBahamas, D!esty MarmeAssociates in the FloridaKeys, Aquaculture Development Ar'S in Denmark,and Instant Ocean Hatchery in central Florida,

C-Quest.Inc. completed construction of a 15,000ft marine6sh hatchery in 1991.The hatcherycontams twelve hundred 40 gallonaquariums used for broodstock.larval rearing, andresearch. The f'acilitieshave since been exgeaded to includefour hundred300 gallon outdoorlarval rearing and grow-outtanks, nearly doubling the sizeof the original hatchery.In addition,there are four 25.000gallon concrete ponds used for large-scale cultureof microalgaeand grow-out of brineshrimp. Filtration includes biofilters. sand filters.cartridge tihers, diatomaceous earth Glters, and UV sterilizer>. CommeriMproduction efforts initially focused on clovm6sh,but have since included two speciesof gobyand three species of dottyback.Several other species have also been culturedat the C-Questhatchery. The list of speciescultured by C-Questincludes the following:

Species Common Name Amblvgobius rainfordi ' Rainford'sgoby A mphipnon akallopisis Skunk clownfish A mphipnon akindynos Barrier Reef clov~

PREPARED BY GADTC HNAL REPORT - CULTURE OF MARINE AQUARIUM FISHES

Amphiprion bicincrus Two-banded clownfish Amphiprion clarlai Clark's clovndish Amphiprion ephippium Red saddleback clownfish Amphipnonfrenarus Tomato clowafish Amphiprion melanopus Cinnamon clownish Amphiprion ocellan's Common clovIm6sh Amphiprion percula Percula clownfish Amphiprion perideraion Pink skunk clovnfish Amphiprion sandaracinos Orangeskunk clownfish Calloplesiopsaltivelis ~~ Comet Chrysipreracyanea '~ Orange-tailDamsel Gobiodon cirn'nus Citron goby Gobiodon oki~e ~ Okinawa goby Gobiosorna evelvnae Sharknosedgoby Gobiosomagenie Genie'scleanmg goby Gobiosoma macrodon Tiger goby Gobiosomaoceanops Neon goby Gramma lorero ' Royal gramma Gramma melacara ~ Black cap basslet Ophisrognarhusaurifons ~ Yellowheadedjawfish Premnas biaculearus Maroon clownish Pseudochromis duroiri Neon dottyback Pseudochromis flavivertes Sunrise dottyback Pseudochromisfiidrnani Orchid dottyback Pseudochrornispaccagnellae ' Royaldottyback or Bicolor dottyback Pseudochromisporphyreus ~ Magentadottyback Pseudochromisspringeri ~ Sprmger'sdottyback Prerosvnchiropussplendidus ~~

' Occasionally available. " Not likely to be conunerciallyavailable in the near future.

In additionto the specieshsted above. C-Quest has produced one hybrid variety of clownfishthat hasresulted Som &re accidentalpairings of Amphiprionocellaris and A. percula.The hybrids were reared without problems and are called "Percullaris" clownfish. C4Iuesthas also developed or culturedseveral color varieties or strainsof someof the clownfishspecies. A brightorange-red A.percula and a fastdeveloping dark stram called Earlyblack A. perculahave been developed. A strain o fA. clariii calledWhite-backed A. clarkii containsa iocabzei albino areawhich resemblesa white saddleon the fish'sback. Another strain called Heart-sleevedtomato clownish is an A. frenatus with a white dot at thebase of oneor bothpectoral 6ns. C-Quest also produces both white-stripe5 normal coloration!and blue-stripedCinnamon clos+fis A. melanopus!,and white-striped normalcoloration! and yellow-striped Maroon clownfish Premnas biaculearus!.

10 PREPARED BY GADTC FINAL REPORT - CULTURE OF Iver/~ AQU/dtHM FISHES tt

C-Questis now the largestcommercial hatchery for marineaquarium fish in the worM. Thecompany is ableto sellev~~g it produces,and is in theprocess of developingan arrangementto marketits fish througha largecham of pet storesin Californiacalled Pets Mart M. Moe.per. comm., 1996!. Pets Mart plans to marketonly tank-raised specimens of marineorgani!tms, instead of wild-caughtanimals. C-Quest has about 50 wholesale customersand sells about 10,000 fish per meath. Dottybacks Pseudochromissp.! accountfor onlyabout 150 fish per month. although the company has been getting $15 to $20per fish W. Addison,per. comm., 1996!. However, wild-caught dottybacks have startedbecoming more available as new Red Sea collecting stations have recently opened, andthe price of thesewild-caught dottybacks ismuch lower than the tank-raised fish. As a result,C-Quest is beginningto havesome dif5adty in maintainingits price for the dottybacks.The exception is Pseudochromisdutoiti, which has not been collected in large numbersRom the wiM at this time W, Addison,per. comm.,1996!. The ownerof C- Quest,Bill Addision.is planningto goto SouthAtrica in thenear future to investigatethe possibilityof a joint venturewith an investorthere who claimsto havesuccessfully culturedClown triggerfish. Emperor angelfish. Moorish idols, and Powder blue tangs.

ReefPropagations. founded by Joe Lichtenbert in April 1990,is a small00 squarefoot! buementoperation located in theChicago, IL area.The company began producing their first salablefish in October1990, and by the endof 1995had soldabout 50,000 fish for morethan $150,000.About one third of thesefish were purchasedRom C-Quest and resoldby ReefPropagations J. Lichtenbert,per. comm., 1996!. Sales per week averaged 70 fishfor the first two yearsand rose steadily to over250 per week Bof In Prep.!. Productionhas recently dechned due to technicalproblems. Total cashoutlay at the beginnmgof 1995 was about $53,000 of whichabout $10,000 was allocated for research anddevelopment Boff, In Prep.!.'Hm operationnow hasa positiveoperating revenue andis operatedby 1.5people with a 45hour work week. A companyin Utahcalled DMS Fisheriesis using marme geothermal water to raise severalspecies of marineaquarium fish. The company was started in early1994 and is commerciallyproducmg several species of clownfish,including AInphiprion frenatus. A. melanopus,A. ocellaris, and A. penta. DesertFisheries has also successfully reared smallquantities of thedottybacks Pseuabchmmis flaviv~cr andP. Iridntani.At the researchstage, the company isspeamng the goby Alnblygobius phalaena. although larval rearingattempts have not been success' at thistime. Other species that are targeted for researchare the Coinet and someangeifish species.

Informationobtained irom an advertisementin an aquariummagazine indicated that a tropicalmaitine fish hatchery in London,probably Tropical Marine Centre Ltd.. has bred thefollowing, species in commercialquantities usmg the artificial sea salt Tropic Marin: Amblygobiusphalaena, AInphiprion allardi, A. clarjai.A, frenatus,A. melanopus,A, oceliaris.A. percula.Gobiodon olanmvae, Gobiosoma evelvnae. G. oceanops,G, puncticuiatus,Hippocampus comes, H. fuscus. H, hystrix.and Lvsmara debelius Fire Shrimp!.

PREPARED BY GADTC FINAL REPORT - CULTURE OF MAMNE AQUARIUM FISHES ~ I

Red SeaFish pHarm Ltd. is a manufacturerof arti5cial seasalt Coral Reef SeaSalt! and other aquarIIumproducts in Israel 'I%is company began an intensive Researchaad Developmetttprogram to develop new methods far artificial propagation of marine ornamental ashes in 1994, aad has successfully cultured the following species: Amphiprion bicinctus. A. clarkii. A. frenarks. A. ocellaris. Chrysipt'era paraserna. Ophistognathusaurifons, Calloplesiopsaltivelis, Psecdochrnmisfridmani, P. jlaviverzex, aad Pterosvnchiropussplendidks Broas, 1995!. Red Sea Fish pHarm was the first companyto breedtwo speciesof 6sh ie., Pseudochmmisfridmani aadP. flaviverrex! that are indigenousonly to the Red Sea. The aqmiriumsaad larval taaks at the experimentalhatchery are arnmged in a closedsystem equipped with biologicalSters aad proteinslmmer3. Although the facility is locatednear the Red Sea,artificial seawateris used in the culture systems,The hatcheryhas separatefacihties for the culture of microalgaeand zooplankton.The main goal of the researchprogram is to developthe knowledgeaad expertise to establisha comnierciaihatchery producing a widevariety of coral reef fishes. However, the companyis also usmg this researchproject to promote their arti6cial sea salt. Red SeaFish pHarm is presentlylooking at the economicpotential of a commercialhatchery facility M. Moe, per. comm., 1996!. The companybeheves that all the speciesthey have raised have commercialpotentiaL except for Chrysiptera parusemaand Pterosynchiropussplendidus. The formerspecies does not havea good marketvalue and the latter specieshas a very low fecunditywhich limits production Brons, 1995!. The hatchery operation is still at the researchaad developmentstage, althoughthe companyhas reportedly marketedsome fish mto Europe S, Browa, per, comm., 1996!.

SeaPhiz formerly EcoActivity, Inc.! is a relatively new company spemhzing in the manufactureand operationof spxiabzed homeaquaritim systems model ecosystems!the companycalls Ecotariums aad wildlife maricutturesystems called ecoCulture facihties The ctiiture systemsare recirculating hatcherysystems using algae scrubbertechnology developedat the SmithsonianInstitue. SeaPhizctnreatly has oae ecoCulture system operatingin Cleveland,Tennessee and anotherin Terahaute,Itzhaaa. Species currently cultured in these hatcheriesinclude Amphiprion clarkii, a black strain of A. perckla. Gobiosornaoceaaops, Gmmnuz lore', Grtxmmamefacara. Hippocampussp., PseudoehromisChtoiti, P. flaviverter, andP. Iridmani. At thepresent time. thel facilities are just beginningto get up to commercialscale production. with oae or two active breediagpairs of eachspecies aad a productionof about200 to 250 marketable6sh per month J. Walch,per. comm.,1996!. The Arizona-basedcompany is alsoexperimenting with the ndture of Valencienneastrigal. In addition to the culture of marineaquarium fish,the company is alsoculruring live coral,live rock, aad live sand.

AqualifeResearch Corp., locatedin Walker'sCay, Bahamas.was the oldestcompany producingmarine ornamentals. Until therecent expansion of the C-Quest.Inc, hatchery,it was also the largestmarine ornamental fish hatcheryin the worM. AqualifeResearch Corp.closed its hatcheryoperation in January1996, after operatmgon Walker'sCay for the last 12 years.The fish inventory aad equipmentwere sent to Harbor Branch OceanographicInstitute ia Fort Pierce,Rorida, where a newinvestor Dick Williamsf'rom

12 PREPARED BY GADTC FINAL REPORT - CULTURE OF M ~K AQUPdGUM FISHES

Palm Beach,FL! is beynaag to investigatecotnmmial marineaquarian fish culture G. Waugh,per. comm.. 1996!.

Aquahfe's 25,000 ft hatchery and grow-out facility included a broodstock room contammgover 250 broodstocktanks for clownish andneon gobies the goby broodstock were kept in a separateroom that was air conditioned!and a larvalroom containingover 40 largetanks the numberof larval rearingtanks was laterdoubled as productionwas expanded!and severalsmaller glass aquariums for larval culture.The live foods area contained an algae laboratory for maintaining small algae cultures, several tanks for hatchingbrme shrimp, four largetransparent cylinders for outdooralgae culture. six 5-ton tanksfor growingalgae and rotifers. and several large outdoor tanks for brineshrimp. The grow-outarea contained over two hundred300 gallon tanks, Aqualife used saltwater wells which reportedlypiped morethan a m01iongallons of seawater through the hatchery daily. Over 20,000gaQons of microalgaewere producedeach month. The facility was designedto be capableof producingmore than a millionfish eachyear. but the actual productionnever came close to that amount.

During its peak productionin 1989,Aqualife produced and sojd over 20.000fish per month of the following species:Amphipnon clarkii, A. fivnatus, A. - elanopus,A, ocellaris, A. pereula. A. penderaion, A. polymnus.A. sandaracinos,Gobiosoma oceanops,and Premnas biaculeatus both Gold-stripeand regular Maroon clownish!. Aquahfe'scustomers included 42 marineaquarium fish wholesalersin the United States, one in Canada,one in England.and one in West Germany.In 1994, Aqualifewas producing about 10,000fish per month.

In the earlydays of Aqualife972-1983!, severalother species were successfully cultured that neverreached commercial production or werecommercially produced for only a short time: Equetusacuminatus, E. lanceolatus,E. punctatus,Gobiosoma evelvnae, G. multifasciatum,Grumma loreto, G. melacara, Hypoplectrusgemma, H. unicolor, Microspathodonchrysurus, Opistognathus aunpons. Pomacanthusarcuatus. and P. paru. Aqualifealso produced two hybrids;Equetus acuminatus x E. lanceolatus.and Pomacanthus arnustusx P. pam

D!pestyMarine Associates operated a smallhatchery near Marathon. Florida 6om 1983 to 1995.While it operated,the hatchery produced several species of clownfish.the Neon goby,and seahorses, asa supplementto the main part of thecompany's busmess. which is the collection of marinefish, invertebrates.and live rock for the aquariumtrade. Although thecompany no longer raises fish in its ownfacility, it buyssome tank-raised fish Bom C- guest. Inc. ta include in the inventory of wild-caughtfish that Dynasty sells to wholesalers.Species commercially cultured at Dynastyincluded Arnphiprion clarkii, A. Irenatus,A. melanopus,A. ocellaris,A. perideraion,Gobiosoma xanthiprora, G. oceanops,Hippocampus sp., and Premnasbiaculeatus, Other species reared at an experimentallevel includedAmphiprion bicinctus, Amphiprion chrysopterus, Apogon compressus,Bodianus rutus, Chrvsipteracyanea. Equetus acuminatus. Equetus lanceolatus, Z punctatus. Gramma loreto. Microspathodonchrvsurus. and

PREPARED BY GADTC FINAL REPORT - CULTURE OF MARINE AQUAIUUh f FISHES I ~

Opistognathusaunfmns. Dynastyhad also produced hybrids on an expmaeatal basis. includingEquetvs acuminarus x E.punctarus and E. lanceolcuusx E. puncrarus.

At one time. the broodstockincluded about 60 pairs of clownfish aad 48 pairs of Neon gobies.Seahorses were raised Rom wiidwaught fish ca~kg eggs.About two thirds of the broodstock pairs spawnedon a regular basis. Each broodstock tank had its own individual fiitratioa system.but the larval and juvenile grow-out systemswere connected in a semi-recirculating systeaLwhich included a 30,000 gallon reservoir tank where seawaterpumped directly t'rom the ocom was sterilize with chlorineaad dechloriaated before use in the system.A headertaak, rapid sandfilter, biofilters. and protein siwimmer completedthe fiitratioa system.The larval tanks were locatedinside the hatcherybuilding aad the grow-out tanks aad live foods production taaks were located ia a greenhouse outsidethe hatchery.The growwut periodwas approximatelyfour months.

Aquaculture Deve!opmentA/S ADAS! in Denmark, was also producing clowufish commercially, but is no longer in operation. The company commerciallyproduced ten speciesof clownish. using syntheticseawater Instant Ocean! aad recircuiatmgsystems. The main species produced were Amphiprion frenatus, A. ocellaris, A. sebae, and Premnas biaculeatus. Thc company began supplying the aquarium markets in Scaadiaavia,Great BritaiL Germaay,aad the Netherlandswith tank-raisedaaemoaefish in 1990,aad m November,1991, the first Danish-producedcbmnSsh were introducedto the American market Torbea, 1992!.

The hatcheryfacility at ADAS consistedof a broodstocksystem, a larvalrearing system a juvenile grow~ut systemand a hve food productionarea. The broodstocksystem coatamedabout 16,000liters of water that was circulatedthrough sandfilters aadtwo UV sterilizersbefore returningto the 98 spawningtanks. Each 125 lifer spavmmgtank containedan undergravelfilter for biological filtratio. Thc larval rearingsystem contained about 1000 hters of water ia four tanks. %hegrow-out systemhad a capacityof 52,000 litersaad was separatehorn but simihsrin, design to the broodstockSter systemThe live foodproduction system was probably the mostsophisticated part of the facility,with two speciesof microalgae.Rhodomonas aad Isochrysis, grown in two 600liter photoreactors. Algae production was idly automated,with a maxinnundaily productionof 400 1itersof algae. Rotifers were producedin seven taaks, each with a volume of 800 liters, The compaaydeveloped it owndry foodfor grow-outof thejuvemie fish Torben,1992!.

Instant Ocean Hatcheries.founded aad managedby Frank HoE was m operation from 1974to 1984, producingclowafishes aad neon gobiescommercially at an mlaadhatchery ia centralFlorida. Pamualproduction of juvenilesreached over 250,000using Instant Ocean artificial sea salts in a closed cuing system Species produced included: Amphiprion akallopisos,A. cl'arkii, A. ephippiuIn,A. frenatus. A. ocellaris. melanopus.A. persia. A, polyInnus,Gobiosoma oceanops, aad Premnas biaculeatus. Ia addition.Instaat Ocean produced three hybrids: A. frenatusx A. ephippium.A. frenarusx A. melanopus.and A. ephippiumx A. melanopus.The hatchery was aa 11,200ft 2 split level building that contained a 10.000 galion larval rearing system. a L2.000 galion

PREPARED BY GADTC FINAL REPORT - CULTURE OF MAIUNE AQUPdUIZ41 FISHES broodstocksystem and a 24,000gallon grow-out system. The larvalrearmg system contained80 forty gallonaquariums. The broodstocksystem contamed 180 plywood tanksof 29 or 40gallons. The grow-out system contained 384 forty gallon plywood tanks. Theaquariums in each system were connected to reservoirs and filters. The facility also containeda 10,000gallon experimental system used for growmgalgae, invertebrates and extrapairs of subadultfish. There were ten annal 4,000gallon reservoirs and six 10,000gallon outdoor recychng ponds. Approximately 70'/o of thewater used in the hatcherywas recycled Hoff In Prep!. Someresearch has been conducted by a fewuniversities, pubhc aquariums. and private researchinstitutions on the cultureof marineaquarnnn species. The University of Hawaii, in a researchprogram under the direction of Dr.Chris Brown, was successful m rearing twospecies of damsel53h,Dascyllus albiseia and Dascyllus an>anus, to the juvenile stage,with one of therearing triaLt resulting m over40'/o survivaL Larval feeding during theQrst two weekspost-hatch was accomphshed by maintaming a continuous indoor cultureof algae Tetruselnis sp.! androtifers in the larvaltanks. supplemented by progressivelylarger wild zooplankton Danilowicz aad Brown. 1992!. Dr, Brownis presentlyworking on developing methods for captive spwNmng of Centropyge fisheri, C. loriculus,and C potteri C. Brown,per. comm., 1996!. HarborBranch Oceanographic Institution HBOI!, Fort Pierce, FL beganan ornamental msrinefish andinvertebrate culture program in 1993.To carryout this reset, the Instituteequipped a 450ft glassgreenhouse with three independent seawater systems, eachcontaining four 40-gallon aquaria, a wet/dry biofilter and foam fractionator, a GAC contactor,and UV sterilization.The facility also houses translucent fiberglass columns for phytoplanktonand marie zooplankton production, and four 3000-hter recmmhting larvicuituresystems. HBOI has successfully reared larvae of theBanded coral shrimp, Stenopushispidus. through metamorphosis after102 days and the Peppermint shrimp, Lysmatavecndemanni, after35 days. In addition, theSergeant major, AbudefduIsaxanlis wassuccessfidly cultured. The Institute is alsoconducting research on the following spccÃ$:

Species Commoa Name Lysmatagrabhami Scarlet lady Periclimenespedersoni Pederson'scleaner shrimp or Caribbeananemone shrimp P. yucatanicus Clown anemoneshrimp Stenopusscuteilatus Goldenbanded coral shrirrrp TheUniversity of Texas recently began a research program to investigate thecu1ture of marineornamental fish and shrimp. Under the direction of Dr.Joan Holt, the following specieswere spawned m the laboratory: Bodianus pulchellus. Centmpyge argi,Equetus umbrosus,Halichoeres caudalis, H. maculipinna.I vsmata Murdemanni, Serranus tigrinus.and Thalassoma bifasciatuln. A largetank ie.. 1000gaHon raceway! was requiredto get adult B. puichellusto spawn. However, larval rearing has not been successfulforany of the fish species, except B.pulcheilus and Z. urnbmsus.dueto a hack

15 PREPARED BY GADTC FINAL REPORT- CULTUS OF M,CRINEAQUARIUM FISHES I ~ af a suitablefirst foodfor thetiny larvae.Rotifers are too largeas a firstfood for these species.WiM plankton was also used without success for someof the fishspecies. However,the laboratoryhas been more successful in culturingthe Peppermimshrimp Lvsmatai~cntenanm' through the larval aad juvenile stages. The Pg~mmrt shrimpcan bereared through the larval stage in 30 to 50 days,depending on watertemperature and otherfactors. High waterquality aad the useof enrichedAnemia are importantfor successAdlycuitttriag this species J. Holt, per.comm.. 1996!.

The Universityof Walesbegan a researchprogram in 1993to rear severalspecies of cleanershrimp. Dr. DavidFletcher. prmcipal investigator, has succeededin rearing Lysmataamboinensis. L debeliusaad Stempus hispid', with the culturedjuveniles now producinglarvae in thelaboratory D. Fletcher,per. comm., 1994!. He is aowdevelopiag techniquesfor the massculture of the two Lvsmataspecies, aad refinements ia rearing techniqueshas reduced the larvalperiod for thesespecies fi'om 13 or 14weeks to seven ~eeks D. Fletcher,per. comm., 1996!. Of thethree species listed above, S. hispidusis the raostdifficult to culturethrough to thejuvenile stage. Rotifers are aot requiredas the first feedfor thisspecies, aad Dr. Fletcherhas had success with a varietyof live aadinert diets. In particular,a aematodespecies enriched with marinelipids and pigments was used to supplythe essential nutrient pro5ile for thesecrustaceaas D. Fletcher,per. comm., 1994!.

Dr. Fletcheris alsoinvestigating autritioaal requirements of clownish,including vitamin C requirementsia Amphiprion clarhi Fletcherand Wilson, 1996!. He is coasultiagwith a privatecompany to developaew diets for clowafishaad other marine aquarium fish. He is alsoconsulting with anotherprivate company to developcommercial rearing methods for otherspecies of marineaqmlrium fish, inchding tangs. The companyhas been successful ia rearingthe Catalina gaby, Lyly pm' daNi,aad may be the first to ctdhrethis species throughthe hrval stage,They have also been very successfidm rearinga varietyof seahorsespecies, producing a secondgeaenttion injust 7 to 11weeks. Improved nutrition hasbeen an important factor ia thesemess with seahorses.This group is alsodeveloping aewhve feeds for specieswith very smalllarvae i.e., hrvaethat caanotconsume rotifers at first-feeding!and new technologyfor iaducedbreeimg D. Fletcher,per. comm.. 1996!.Emphasis is also being placedon improvmgbroodstock aad larval nutritionto achievegreater successwith marine aquariumspecies that have been difficult or impossibleto cuhmem thepast.

Variouspublic aquariums have in the past or are presentlyconducting research on the cultureof maine aquariutaspecies. The WaikikiAquarium. affiliated with the University of Hawaii,conducted small-sca}e research on culturingmarine aquarium shrimp. Under the directionof SydKrauL the Aquariumsuccessfully developed rearing methods for the Harlequinshrimp, Hyntenocera picta. Usingenriched Artemia, the Aquariumwas able to rear thous'Mtdsof juveniles.The Aquariumis not presentlyinvolved in researchon the culture of marital ornamentalfish or shrimp,but Mr, Kraul is planningto resume cotnmercialculture of the Harlequiashrimp later this year S. KrauLper. comm..1996!. TheBerha Aquarium has been culturing several species of clownfishand seuiag them to raisefunds for variousprojects at the Aquarium C. Brown. per. comm., 1996!.The

16 PREPARED BY GADTC FINAL REPORT - CULTURE OF MASSE AQUAIGLM FISHES

Aquariumis alsoconducting research with otherspecies of marineaquarium fish. The BaltimoreAquarium is successfullyculturing the Highhat,Equetus acuminatus and the Veongoby, Gobiosoma oceanops. The TulsaZoo hascultured the Atlantic spadefish, Chaetodipterusfaber, andspawned several other species.

Severalspecies af marineaquarium fish andshrimp are now being cultured on a smail scaleby marineaquarium hobbyists, and many of thearticles oti rearingmarine ornamental speciesin the aquariumhobby literature were written by individualhobbyists see Appendix2!. MartinMoe, the first personto commerciallyculture marine aquaritum fish andthe founder of AqualifeResearch Corp., is currentlycuittuing the Orchid dottyback. Pseudochromisfridmani,in an experimenta culture system at hishome. He reportedthat the adultsbegan spawning before they were a yearoid. andproduce a spawnof about 1000eggs every 5 or 6 days.The larvae can eat rotifers when they first begin feeding, and canbegin eating Artemia nauplii within a fewdays M. Moe,per. comm., 1996!.

Objective2. Literaturereview A surveyof theaq~ium hobbyliterature was conducted for mformationan spawning and rearmgmarine aquarium species of fish and shrimp.Information on culturee8'orts by marineaquartum hobbyists has recently been collected and orgamzed into a quarterly newslettercalled '"Hm Breeder'sRegistry." In acbhtionto articlesin the newsletter,the editorshave also formed a computerdatabase of spawmng and remng reports for marine aquariinnfish and invertebrates. Other aquarium literature containing reports of culturing marineaquarium species include the newsletter"SeaScope" and various aq~ium magazines.A reviewof thesciet~c lit

Objective3. MicronesianSpecies List:

Species Common Name Acanrhurus leucosternon + Powder-blue tang Amblygobiusphalaena ~ Bandedgoby, Brown-barred goby, Dragon gaby Amblvgobiusrainfordi ~ Rainford'sgoby Amphiprion bicinctus ' Two-banded clownfish or Red Sea clownfish Amphiprion chrvsopterus~ Orange-finor Blue-stripeclos~ Amphiprion clarkii ~ Clark's ciownfish Amphiprionephippium ~ Red saddleback clownfish Amphiprion Inelanopus ' Cizuunon clownfish Amphiprion peri deraio ' Pink skunk cloddish Balistoidesconspicillum + Clown triggerfish Calloplesiopsaltivelis ~ Comet or Marine betta Crvptocenrruscinctus ~ Yellow prawn goby Gobiodon citrinus ' Citron goby Gobiodon okin~ue ' Okinawa goby

17 PREPARED BY GADTC ANAL REPORT - CULTURE OF >r&dUNE AQUMKM HSHES I ~

Hoplolatilus starclti Purple-headedsand tilefish Meiacanrhus atrodorsalis Lyretail blenny MeiacantltusgramInistes ' Stripedblamy .Vemateleotris decora '~ Decorated dartfish NeInateleotris rnagnifica Fire dartfish or Fire goby Platax orbiculans + Circular spadefishor Roundbatfish Platax pinnancs Pinnatespadefish or Redfinnedbatfish Plauu teira Longfinspadefish or Teirabatfish PoInacantltusimperator + Emperorangeifish Pseudochrornt'sporpkyreus ' Magentadottyback Ptereleotns zebra ~~ Zebra goby Sigantts Lo! vulpinus Foxface rabbitfish Signigobiusbiocellatus + Signalgoby or Two-spotgoby Valencienneapuellaris Maiden goby Valeneienneastrigata " Blue-Streak Goby ZancltIs corrmtus + Moorish idol

Successfullycultured in captivity. " Spawnmg in captivity. + Reportsof thisspecies bemg successfully cultured need to beverified.

Objective4. Culture Methods Twomated pairs of theCmnamon cloth, AmphiprionmelanopNs were obtained by caanulatingseveral individuals aud examining the samples to determine the sex of thefish Figures5 and6!. Matureeggs or spermwere found in thesefish. The sex of someof the fishsampled could not be determinedby canmdation.Three pairs of A. vrelanopus spawnedduring this study, for a totalof 2l spawns Table l.!, However,the majority of thespawns were produced by one pair. A secondpair was lost early in thestudy when one of the fishjumped out of thetank. A thirdunidentified pair m atank containing several clown6shspmmed one time, but waslater accidentally disturbed durmg routine maintenanceand did not spawnagain. No pajr formationwas observed and ao spawumgs occurredin the 10-tontank containmg the large group of A. Inelanopurduring the 3 to 4 monthsthat the 6sh occupied the tank. Also. no spawning occurred inthe other clowufish speciesover a severalmonth period. Spawnsize increased during the study period 6'om less than l00 to nearly500 eggs per spawn.Some spawns were not used for larval rearing trials. Clownfish eggs hatched in 8 to9 days,depending onwater temperttture ambient ~ater temperatures during the project rangedfrom 27 to 29 C!.The eggs elongated and became darker as the embryos developed,The eyes developed andthe yolk sacks became smaller shortly before hatching Figure7!. Newly-hatched larvae were 4 to5 mmm length.Larval survival in rearing trialswith AInphiprion Inelanopus ranged from less than 8'/o to over90'/o Table 2!. Larvalmortality occurred pritnarily during the first few days aAer hatching.

18 PREPARED BY GADTC j -i FINAL REPORT- CULTUS OFMARINE AQU&GUM FISHES ~ ~

Amphiprion melanopus Cinnamon Ciownfish

Pair T-7 Pair T-8 Pair T-17

Hatch Date Hatch Date Hatch Date 12/23/94 12/16/94 04/20/95 01/10/95 01/22/95 01/31/95 02!01/95 02/25/95 02/13/95 05/03/95 05/12/95 06/01/95 07/26/95 10/14/95 10/25/96 11/06/95 11/18/95 11/30/95 12/31/95 02/04/96 02/27/96 ' Note: Egg massdied beforehatch

Table1. Spawningfrequency ofthe Cinnamon clown6sh, Arnphipnorr melaaopus.

20 PREPARED BY GADTC <~p:j>j<'< ' < [;,'~i I '[ Xl,. p,JX'',>. ll NATAL!' 14

' '[ q[~- r r q~,g,' .',il,~~<'I!lg vr:,,r." =l,~c!>1'KcQ' ED=DJL'h'~<".

p~p.

O o~OW O O F 2 9' B N '4 w~ 'LA K QPJZ ooo> 0 o ~

V A ILW CO X a+ CO

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00 + ~ W w 0 Ha ~a -xg~e~xc 0 Qs g Q ~~~Wgg~g~>' IQ ~ W 0

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gg FINAL REPORT- CULTURE OF IWBUNK AQUABZUMFISHES

Juvenilemortality was less than 5 ' duringthe study period. No seriousoutbreaks of diseasewere observedduring this time. Clo~mfishreared in the studyreached a marketablesize at about3 to4 cmtotal length Figure 8!, Growthof juvenilesia thefirst halfof thestudy was very slow, with only about 20'/o reaching a marketable sizeafter six months.Faster growth was obtained for a laterbatch of juveniles, with over 50'/o reachiag a marketablesize after four months.About 100clownfish were marketed to a localpet store aad aaother55 fish were marketedon a trial basisthrough a local fish collector/exportertowholesalers in Hawaii and. Los Angeles. The incidence ofdeformities wasvery low < 5'/o!ia most of the batches of clowafishraised at the GADTC. However, a higherincidence ofdeformities >35/o! occurred injuveniles &om a coupleofthe latter spawns. Thepair of Blue-streakgoby broodstock began spawning about one month after bemg purchasedBom a localpet stare. Initial "-pawns were noted when larvae were discovered ia thebroodstock tank at night. In theb gmamgof thestudy, the pipe used as a spawning sitewas not examinedfor the presenceof eggsfor fearof disturbingthe spawnmg behaviorof theadults. However, it waslater determined that the spawning pipe could be exammedona regularbasis without disrupting spawning and parental behavior. During spawning,anegg mass was attached tothe inside of one of the pipes Figure 9!. Artificial incubationof later stage eggs was attempted, byputting the pipe containing theeggs into a largebeaker, with an airstoae placed to providebubbles flowing over the eggs. Another methodof artificialincubatioa was to placethe eggmass in a screenedopea container submergednearthe surface ofthe larval tank with an airstone placed next to the eggs. The eggswould stay at the bottom of thescreened container and the newly hatched larvae wouldbe carried over the edge of thecontamer and into the larval tank. Hatch rates of approximately50'/oto 90'/o were obtained when artificial incubation wasattempted, Developmentof an egg mass was observed under the microscope. Newly laid eggs were yellowand spherical inshape, forming a large mass orcluster that extended outSom the substrate,Asthe eggs developed, light yellow larvae could be seen in a clearoval shaped eggsack The eggs sacks were attached toa strandatthe tail end of the larvae and the eggmass was similar inform to a grapecluster, The larvae were initially straight. but as thelarvae grew, the tail began tocurve back toward the head to form a "V'shape Figure 10!.Larvae were well-developed athatching, with pigmeated eyesand what appeared to bea hllyformed gut. Only a smallamount of yolk remained athatching. The newly hatchedlarvae were lessthan 2 mm in length. Fecunditydata was not collectedduring the study,although some egg masses were preservedforlater examination. However, it appeared thategg masses contained about 2,000to 5,000eggs. The incubation period was 4 to5 days,with eggs hatching early in theevening. The interval between spawns i.e., hatching dates! was variable. ranging f'rom 9 to 24 days Table 3!. Oae spawn may have gone undetected early in thestudy. Spawningoccurred eleven times f'rom August 14" -' to April 1995 for aa average ofabout onespawn every 17 days. Two gaps in sp; nmgmay have been due to missed observationsand were not calculated into the a~ erage spawning interval. Shortly after the

23 PREPARED BY GADTC .< .P~ l-" ' .:.. l l k ;k.-P R i .;',;!. I l,P~ ;l >l.kR.';;:= ' l!LAl .[L,Xf. Fl-i I.~ FINAL REPORT - CULTURE OF MAPLE AQUAIUUM FISHES

Ptereleotns zebra Valeneienneastrigatas ZebraGoby Btue-streakGoby

SpawnDate Hatch Date SpawnInterval 11/06/94 0 08/17/94 0 11/19/94 13 08/27/94 9 12/02/94 13 09/07/94 11 12/17/94 15 09/24/94 17 01/15/95 ? 10/24/94 ? 01/26/95 11 1 1/09/94 16 02/04/95 9 11/28/94 18 02/14/95 10 12/18/94 19 02/25/95 11 01/11/95 24 03/10/95 13 02/04/95 24 03/21/95 11 04/04/95 ? 03/31/95 10 04/15/95 15 Average 17.25 04/25/95 10 05/05/95 10 05/16/95 11 05/27/95 11 06/06/95 06/15/95 109 9 06/24/95 ~ Note:Spnm intervals aredays between sprwas. Intervals 07/04/95 108 07/12/95 with a "?" indicatean unknowninterval due to a possible 07/25/95 13 missedspvammg observation. 08/06/95 12 08/16/95 10 09/15/95 ? 09/29/95 14 10/13/95 14 10/27/95 14 11/09/95 13 11/19/95 10 12/02/95 13 12/19/95 11 01/17/96 ? 01/27/96 10 02/08/96 12 02/19/96 11 03/01/96 11 03/13/96 12 Average 11.57 Table3. Spawningkequency of gobiesat theGADTC.

26 PREPARED BY GADTC FINAL REPORT - CULTURE OF 1VbVUNK AQUAIVLM FISHES

1astrecorded spawnmg in April of 1995,the pair were moved to anothersmaller tank with someother fish becausethe tank theywere in wasneeded for anotherproject, and other suitabletanks were not availableat the time. The pair did not spawn after being moved andeventually died. A few rearingtrials were conducted with Blue-streakgoby 1arvae, feedingthem s-type rotifers. However. none o fthe 1arvae survived more than a fewdays.

Spawnmgof thezebra goby broodstock pair occurred on a regularbasis throughout the study,beginmng about two monthsafter the pair was acquirecL Spawns were found early in themorning, and hatching occurs imnxdiateiy after dark on thefourth night. The egg masswas attachedto the underiide of the chm shell and appearedto be aboutthe same sizeas the Blue-streak goby egg valses Figure11!. Embryonic development app~% to be sunilarto that of the Bhe-streakgoby. The eggs could be successful1yhatched in the larvaltank by removingthe shellcontaining the eggsfrom the broodstocktank shortly beforehatching and phcing it in thelarval tank. However, if theeggs were taken Rom the parentstoo soon.hatchmg was usually unsuccessfuL even when the eggswere well aerated.A totalo f39 spawnswere produced in a 16month period for anaverage of about one spawnevery days 11.5days Table 3!. Severalrearing trials were conducted with the zebra goby 1arvae. SS-type rotifers were fed to thelarvae and attempts were also made to feedthem oyster larvae and an artificial diet. However,the larvae did not survive past the first few days when fed ss-rotifers or artificiai dietsand oyster larvae could not be obtained in sufficientquantities when required to feed the goby larvae, Thethree pairs of Bandedcoral shrimp began speivning about one to two monthsafter beingpurchased from localcollectors. Five ah&anal pairswere obtained later in the studyand also began spawning. Balded coral shimp spivwexf i.e., larvaehatched! at intervalsof 13 to 23 days Table4!. Somelarger periods of timebetween spawns is believedto bedue to missingoteenqLtions of bKtching larvae Eggs usually began hatching about3 to 4 hoursafter sunsetand hatching was completedwithm an hour. Spawnsize usuallyranged from 1000to 3000larvae, but spawnsize may reach as highas 5000 larvae.Limited data from one pair suggeststhat fecunditydid not declinewith timetn capttvlty. Severallarval rearing trials were conducted with the Banded coral ~imp Table5!. In the beginningof most of thetrials, rotifers were maintained in the larval tank at a densityof 10 to 20rotifers/ml and the green alye Nannochlompsisoculara was added daily to maintain a concentrationof about 500,000to 1,000,000cells/ml in the larval tank. High hrval survivalwas usually obtained through the first three weeks. Then the diet was gradually changedfrom rotifers to Artemianauplii and survival began to decline.In thebest rearing trial,some larvae survived until day51. Contaminationof hrvai tankswith growthsof hydroidsand small free-mvimming jellyfish medusae sometimes occurred, especiaiiy after larvaewere reared in the sametank for a fewweeks. Total mortalityof the Bandedcoral rkmnplarvae usually occurred within a fewdays after their appearance in large aumi:ers.

PREPARED BY GADTC j4 g' ' '".';;< " ' c i ' L:4L'

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CV 2 P- o FINAl. REPORT- CULTURE OF MAPJNK AQUMUUlvf FISHES

Stenopus hispidus Sanded Coral Shrimp

SCS ¹1 Spawn Spawn Interval Interval Spawn Hatch Date Days! Hatch Date Days! Hatch Date Interval 08l04/94 07/19/94 0 08/25/94 08/27/94 0 23 08/04/94 16 09/09/94 0 15 10/13/94 08/22/94 18 1Q/10/94 10/29/94 16 09/08/94 17 10/24/94 14 11/14/94 16 09/27/94 19 11/09/94 16 11/29/94 15 10/28/94 11/26/94 17 12f15/94 16 11/18/94 21 12/16/94 20 01/07/95 23 12/09/94 21 01/07/95 22 0'I/23/95 16 02/I/95 ? 01/22/95 15 02/11/95 19 Q2l21/95 16 02/09/95 18 02/28/95 17 03/09/95 16 02/27/95 18 04/29/95 03/25/96 1B 03/12/95 13 05/17/95 ? 18 06/19/95 04/14l95 06/03/95 17 08/07/95 06/Q2/95 ? 18 07/31/95 08/23/95 ? 05f1 8/95 16 09/24/96 ?18 ? 06/05/95 18 10/10/95 Average 17.8 07/04/95 1 $29/95 19 07/20/95 1B 11/23/95 08f07/95 18 08/22/95 Average 17.8 09/24/95 10/11/95 ? 17 11/13l95 ?

Average 16.8

Note: SpawnIntervals with a W Indicatean unknown intervaldue ta a passiblemissed spawning observation.

Table4. Spawningfrequency of the Bandedcoral shrimp.Stenopus hispidus.

30 PREPARED BY GADTC FINAL REPORT - CULTURE OF MPdUNE AQUARIUM FISHES t ~

A few earlyattempts to spawnnewly collected sea urchins were relatively successful in producingferti1izgi eggs and larvae.However. the use of seaurchin larvae for larval rearingof gobieswas later abandoned because it becamevery dif5cult to mducespawning in the sea urchins heM at the hatchery.Dif5culties were also eacouateredin spawning oysters,

Objective5. RegulatoryConsults

In 1987Germany hmaed the importatioaand sale of butterQyfish,angelfish. and Moorish idoh. However. determinedGerman hobbyists can still drive acrossthe border to Holland and obtainthese species R. Meyers,per. comm., 1996!.Nature conservation laws in Israelforbid collectionof reefsl.'cazaens entirely, aad the exportof wiM caughtRed Sea fishesis restricted in most other countries oa the Red Sea Brons. 1996!. Hawaii has bannedthe importationof scorpioafishes,eels, aad sharks R. Pyle,per. comm.,1996!. Floridahas size lixnits and bag hnits for marineaagelfish collected Born its waters,The Commonwealthof the NorthernMamma Islands banned aquarium fish collectionin 1992. TheRepubhc of Palaubanned the collection aad export of tropicalala~ aquariumfish ia May 1993 Baker, 1993!. This baa was the result of TheNature Conservancy efForts to protectthe coral reef habitatsof this islandgroup. The U.S. Territoryof Guamis preparmgto banthe exportof wiM-caughtmarine aquarium species from its reefsand lagoons.Local aquarium stores and private citizens wouM still beallowed to collectwith government-issuedlicenses after the regulations are adopted, but wouM not bepermitted to exporttheir catches Baker, 1993;G. Davis,Division of Aquaticand Wildlife Resources,per. comm., 1996!. Captive-bred aquarium specimens couM still be exported under this proposedlegislation, provided the maricuitureoperation receives the certification.In variouscountries mcludmg the Territory of Guam!,selected areas have beendesigaated as mlriae parks or reserves,where the collection of animalsis prohibited.

Objective6. Mariculturecertificatioa of captive-bredrelumine aquanhnn fish and shrimp cultured oa Guam would consig of an ofBcial certi&mte of origin from a mariculture facilityapproved by the Divisionof Aquaticand WiMIifeResources G, Davis,per. comm.. 1996!. Objective7. Cashflow projectionswere developed for a smallclowafish hatchery and grow-outoperation. Table 6 providesstart-up costs for the facility. Tables 7 and8 provide cashflow projections at 16/o aad 4/o interest rates, respectively. Assumptioas used in the analysesare provided in Table9. Themost important factors affectin profitably are consistentproduction of marketablefish, market price aad sales volume, Feed costs are nota majorfactor affectin economic reatrn. although the uait cost of feedfor marine ornamentalfish is muchhigher than commercial diets for foodfish

31 PREPARED BY GADTC ~~>me acea8oaooo g 6 hl OOO g Vl lA lO sh Othe Q~P4g A

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FINAL REPORT - CULTURE OF 14bVUNE AQUAS 'IM FISHES ~ ~

Table9, CashFlow Assutnptions

INCOME l! BalanceForward: Cash Lelance brought forward Rom previous year. 2! Capital:Investtr,eat by owner. 3! Loan:Amount borrowed Sum b«nk or otherBaaacial institution. Assume loan of $90,000for 10 yearsat 16'/o. 4! Sales:Total incomeSam sale of aquariumspecies. 5! Other: Other sourcesof incomesuch as interestearamgs. 6! Total Inflow: The sumof items 1 through5 above. CAPITAL COSTS 7! SitePreparation/WeH: Well cost of $3,000plus $40/ft. for a 100ft welL$3,000 for site clearingand leveling. 8! Tank!plumbing:Broodstock tanks/aquariums; 15pcs. @$100 ea. Hatchery tanks; 8pcs.@ $200ea. Growout tanks; 25pcs. @ $300ea. Plus general PVC plumbing for air «ndwater at $9,000 lump sumestimate. 9! Pumps:2 submersiblepumps for well at $800each and 2-3 smallsubmersible pumps for total cost of $500. 10! Blowers/Aerators:2 bbawers«t $700 each. 1 1!Feace/Lights: Perimeter fence and outdoor security lighting lump sum estimate of $2,500, 12!Pick-up Truck: Purchaseused truck for $10,000. 13!Storage Sheds: 2 shedsat $800each. One shed for storageand one for once/lab. 14!Generator and Elec«I: onegener«tor at $2500and $2000 electrical work. 15!Lab Equipment: One microscope at $1500 and $200 of laboratoryglassware and supphes. 16!Miscellaneous: 10/o of costof capitalcosts exceptfor pick-uptruck!. 17!Total Capital Costs: Sum of items7 through16 above.

35 PREPARED BY GADTC FINAL REPORT - CULTURE OF hVJUNE AQUAIUUM FISHES

Table9. CashFtow Assumptions continued!

OPERATIONAL COSTS 18! Broodstock:20 pairs clownfi!4 with anemonesat $30 per clo~Msh pair plus anemone, 19! Feeds:1st Qtr: $1000 for Artemia and $2,626for Inert feedsfor hatcherybased on start-upassumptions. 2nd Qtr: No feedpurchase req~mi. 3rd Qtr: Cost of feed for broodstockand growwut for one month. 4th Qtr: Cost of feedfor broodstockand grow- out for two months.Yearly thereafter$13,403 for 12 monthsof hatcheryand growout feedsbased on amunptions in "Growout ProductionAssumptions" section. 20! Medications:Lump-sum estimate of $500per year. 21! Equipxnent& Supplies:Cost of algaenutzients, hosing/tubmg/netting/screens, glassware,and buckets/nets. $3,250 per year lump sum estimate. 22! E&tricity: Estimateof $500 per month. 23! 1 lt:S~f l~y R19KI hf I y and$3500/month each year thereafter. 24! Audie%egal/Insunutce:Lump sum estimate of $200per month. 25! Lease:$500/month for 1/2 acresite, 26! Maintenance:Estimated at 44/oper yearof total capitalcosts no costfor 1stquarter!. 27! Contingency:5'/o of operatingcosts each quarter or year. 28! LoanPayments: No paymentfor first quarter,then monthly payments based on a 10 yearloan of $90,000at 16/oand 4'/e interest for tables7 and8, respectively. 29! Taxes:Estimate based on 3'/o of salescosts. 30! Total OperatingCosts: Sum of items 18 through29 above. 31! Total Outflows: Total OperatingCosts plus Total Capital Costs. 32! Net CashBalance: Total InQowsminus Total Outfiows.

36 PREPARED BY GADTC FINAL REPORT - CULTURE OF MARINE AQUAIUUM FISHES

DlSCUSSION OF RESULTS:

Objective 1, C-Queststruggled in the begirmingof its operationwith severalproblems, Including permittmg delays, disputes with governmentagencies. personnel problems. difficulty ia getting quality broodstock.disease aad water quality problems.The company has survivedthe difficultiesof start-up and is now becomingsuccessful from a biological perspective,curving large numbers of clownfish as well as several other high-value species e.g,, Pseudochmnris sp. andthe Comet!.C-Quest is aot yet profitable,but it is expectedto hit the break-evenpoint aadbegin making a profitbefore the eadof the year W. Addison.per. comm., 1996!. The companyhas put emphasisoa researchand developmentof newspecies, and this shouldcoatmue to payoff in the futureand increase profitabilityof the compaay.C-Quest could benefitRom more effective marketing, and higher pricescould probablybe obtainedfor their fish, makingthe companymore profitable.

ReefPropagatioas is a familyowned and operated business, and was startedby a marine aquarium hobbyist without biological trammg or experim~ ia aquaculture. Reef Propagatioashas the advantageof being very closeto a largemarket, and has beenable to get relatively high pricesfor the clowafish it produces probablyas a result of savingson shippiag costs to the customer!. The fish produced have good coloration and the percentageaf saleablefish is nearly 100 percent. Thc companyhas also benefited by buymg some other speciesof clovm5sh from aaother hatchery C-Quest! and reselhng them at a profit, therebymcreasing product diversity aad salesvolume. The capital costs for replicating the culture facihty buiMiag already provided! is less than $10,000. Althoughthis operationis success' financiall, the companyhas had sometechnical problemsthat have limited productionaad profitabiTitym the last two years.These problemswill probablybe resolvedin the near future.The ultimategoal of this small facilityis 450 fishper week with grosssales of 70,000per yearaad anticipated operating expensesof about$10,000 to $1S,000.

Desert Fisherieshas achievedinitial successusing an unlikely source of saltwater i.e., marmegeothermal water in Utah!,for their hatcheryoperation. The water is not the same as natural seawaterin chemicalcompositiotL bemg low in sulphatesaad magnesiumand haviaga low pH. Paamgniais also presentat about 0.24/a.The temperatureof the incommgwater is 90 F. Vaxiouswater treatmentswere tried at one time, but the untreatedwater was found to providebetter results than the treated water L. Nelson.per. comm.,1996!. Aquarium Systems and one or moreother business partners backed out of the businessbecause they dida't think the hatcherywould be successfblusiag the marine geothermalwater. Desert Fisheries has good results culturing some species of clownfish usmgtheir water, but theyhave had difhculty breeding other species Anrphiprion ciarkii, A, penderaionand Premnas biaculeatus!, and this maybe relatedto the differentwater chetmstryof their saltwater.However. the companyis producingseveral hundred marketab1efish per week and obtaining a highmarket price.

37 PREPARED BY GADTC ANAL REPORT - CULTUS OF M ~ME AQUAPJKVf HSHES

SeaPhizbelieves that their technology has the potentialfor culturiag100 to 125species of marineornamental Gsh. The algaescrubber technology used ia theseculture systems is reported to be very efficient at removing ammonia and other waste products. with no water changesbeiag required ia the last 1.5 years of operation J. Walch, per. comm, 1996!. The companyis not selhagtheir hatcherysystem technology at this time. but has decidedto test the facilitiesthey have in operationfirst to evahatethe potentialfar commercial culture of marine ornainental fish and invertebrates.

After more than ten years of operation. +vasty Marine Associatesphased out their hatcheryoperation becausethe companycould aot make a profit raising cloddish in the Florida Keys. This was due manly to the high cost of labor and electricity. Air conditioning was required m the hatchery ia the heat of the summer,and heatiag was requiredin the colder winter months F. Young, per. comm., 1996!.

Aqualife was never profitableand QnaDyclosed down as a resultof cost-cuttingmeasures by the parent company,Precision Valve Corporation, Oae of the mainproblems affecting profitahhty at Aqualife was poor water quahty. The saltwaterwelh that supplied that hatchery bceamecontaminated by petroleum Sam the nearbypower plant, Other wells were later drilled farther away irom the power plant. but the water from these weHs comaiaeda high concentrationof iroa. Aqualife had plans for settingup an intake pipe to pump water directly from the ocean,but the hatcheryoperation was closeddowa before theproject was completed G. Waugh,per. comm.. 1996!,

Whenthe Instant Oceanhatchery operation shut down, the companywas selliagclowafish at about $l.65 per fish, with a prothictioa cost estimatedto be about $2.25 per fish. Severalthhgs contributedto the high production cost. Personnelcosts were high due to a large numb@of employeesand a staF top-heavv ~ matltgement personnel.Salaries were 53.9% of the monthly operatingexpemies. ~ the hatcheryoperations could have beenconducted with half the numberof personnelthat were oa stafF Hoff ln Prep.!. The hatcheryused artificial sea salts ia a recirculating system,but the filtratioa systemused sub-gravelGlters for biologicalfiltration! was not very ef5cientcompared to the filtration technology available today F. Ho6; pers. comm. 1996!. Another problem was that juvenile survival in grow-out was about twice the amount of those fish that were consideredmarketable i.e., overaHsurvival ia grow-out was 43,6%, but 23% of the survivorswere di!cardedas unsalable!.The productioa of a largenumber of fish that were unmarketablegreatly increased the cost of production.

Objective 2. Literature oa mai~ ornanMartalculture is becoming more abundant. although little scientific research has been done ia the past. Most of the available informationoa spawningand rearmgmarine aquarium species comes 6'om aquarium hobby publications.although some research institutions have recently begun research programsoa marineornamentals. Advances ia rearingtechniques for othermarme species caa be appliedto the cultureof marineaquarium fish and shrimp.

PREPARED BY GADTC FINAL REPORT- CULTURE OF 1VbVtINEAQUMUUM FISHES

Objective3. Severalspecies of marineaquarian fish on the MicronesiaaSpecies I.ist have beenreared coaanercially or on an experimentallevel. The Bandedgoby or Dragongoby. Amblygobimphalaena. has been reported to be successfiillyctdtured by a commercial hatcheryin England.Desert Fisheries is alsoattempting to culturethis species.but the smaillarvae will not eat rotifers andthe companyhas yet to get the larvaeto surviveto the juvenilestage L. Nelson,per. comm.. 1996!. A relatedspecies. Rainford's goby, has been cultured in smail quantities by C-Quest.although the larvae are also difficult to rear in largenumbers because the tiny larvaecannot eat rotifers during the first 4 to 5 days. Ciliatesaad copepodnauplii were successfullyused to get a few larvaethrough to the juvenilestage W. Ah%son,per. conan.,1996!. These two specieshave potential for commertM culture ia Micronesia, although they will be more difficult to culture than manyother species.

All the clough specieshave been cultured on a coaanercialor ski-commercial basisby severalcompanies or researchinstitutions, although Amphiprio~ chrysoprerusis reported to be especiallydifficult to get to spawnm captivityand this speciesis probablynot being rearedcommercially at the presenttime Young, 1991A!. Due to its limited availabilityto the aquariumhobby aad strihng appesraacc'.this specieshas a higher market value than mostother clown&A species.Probably the only exceptionis A. latezonatus,which is even lesscommon m the aquariumtrade andhas a wholesalevahie of about$25 per fish.

The Comet has beensuccessfidly cultured by at least three companies,and commandsa high price ia the aquarirummarket Wassiakand Brans, 1990;Brons, 1995!.It is reported to be diflicult to induce to spasm,but the larvae are relatively easyto rear Wilkerson, 1996!. The Citron and Ohnawa gobies have been commerciallycultured by C-Quest Gutierrez, 1996; Wilkmon, 1996!. The Yellow prawn goby has also been reared in captivity and will probably become availablecotmnercially ia the future. The Purpie- headedsand tilefish, HoploIatihu stamki, is uncommonin the aquarimntrade aad ao attempts to culture this species have been reported, but the small size, stunning appesranceand high market value of this speciesmake it worth iavestigatmg.The Striped biennyhas been reared m captivity,and a relatedspecies. the Lyretail blennyMeiaca~thus atrvdorsaris. could probably also be reared, making these speciesgood cmites for commercialculture in Mcmnesia. The dartfish NemateIeorrissp,! are small colorful fish that are popular for mjn~ aquariums.At least oae speciesof dartfish X decora! has spawnedin aquariums,although the eggs are reported to be only about half the size of ciowafisheggs Schilier, 1990!. No attemptsto rear the larvaehave been reported.

The Roundbatfish Platar orbindans is reportedto be under ctdtiv~tionia Taiwan Liao er al., 1995!.A relatedspecies, the Atlanticspadefish. has also been cultured in captivity, andcaptive spawning and larvalrearing are not diflicult Walker, 1991!.No published informationwas found on spawningor larvalreariag of the other two speciesof Indo- Pacificbatfish. However, theselatter species P. pinnums and P. lira! are hkely to have similarrequirements for cultivationand have a muchhigher market value. No published informationwas found oa spawmagor larval rearingof the Magentadottyback. However, this speciesand three other Pseudochrornisspecies have been commerciallyreared by C-

PREPARED BY GADTC FINAL REPORT - CULTURE OF IMMUNE AQU/&JUL FISHES

Quest,Inc. W, Addison.per. comm., 1996!. Adutt P. fridmani begin spawning before theyare a yearold, and produce a spawnof about1000 eggs every 5 or6 days.The larvae caneat rotifers when they 6rst beginfeeding, and can begin eatmg Anemia nauphiwithm a fewdays M. Moe,per. comm.. 1996!. However, larval survival appears to be better whenwild phnkton is provided.and this may indicate nutritional problems that need to be addressed.No reportsof cutruringthe Foxface Siganus vuipinus! werc found, but other rabbit6shof the genusSigunus have been success5illy cultured on an experimentalor commercialbasis, and it is 1ikelythat the Foxfacecould also be cultured.

TheZebra goby Ptereleoiris-ebra! spawns on a consistentbasis at the GADTC,but larvalrearing to datehas not been successtul, due to theextremely small size of theIarvae andthe dif5cuttyin providinga suitabiefood, The Blue-streakgoby Valenciennea strigara!also spawned on a regularbasis at the GADTC, but the larvae of thisspecies is similarin sizeto theZebra goby larvae, and they also have not beensuccessMly reared. 'Hmc gobiesare popular aquarIrum species and have a goodmarket value. It is beheved that successwith both of thesespecies. as well as the Maidengoby V. puellaris! and manyother more highly valued marme aquarian 6sh, could be achieved if oyster larvae or othersimilar sized organisms could be suppiMdon a consistentbasis. Until this happens, specieswith very small larvae are not recommeaded for commercial culture.

Manyother species of connnerciattyvaluable maine aquarium 6sh Som other parts of the world couldbe cuttmei in Micronesiaif their importationfor maricultureis permitted. Theseinclude some of theclown6sh species e.g., Antphiprion frenatus, A. le'eronatus.A, ocellaris,A. pereula,A. polymmrs,and JPreejnas bigndeutus!, most of the dottybacks ie., Pseudochmnrisduteiti, P. flaviverter,P. fn'dmani,P. paccagnellae,and P. springeri!and several other species that have been reared in captivity Appendix 4!, Severalspecies of marineornamental shrimp have been successfully reared in captivity, anda fewspecies e.g., Lysmata debelius and Hymenocem picra! appear to havegood commercialpotential. Although the larval period of thesespecies are relatively long i.e., 35to 50days!, it is manageableandcan probably be decreasei somewhat with improved i hiAi il ii h ~ ii i maybe in producinglarge enough quantities ofjuveniles M. Moe, per. comm., 1996!. Objective4. Broodstock pairing was a problemarea throughout the study. None of the Cinnamonclown6sh Amphiprion melanous! were obtained as mated pairs. but were collectedas groupsof unIMLired6sh. Cannutation to determine the sex of thefish was partiallysuccess6il in the begnnnng, resulting m twomated pairs. However. the sex of manyof the 6sh could not be determined bycannuiation, andthese fish may not have had matureeggs or spermat thetime, The fastest and most reliable method for obtaining matedpairs of aquariumfish for broodstock is to collectnaturally mated pairs fiom the wild. With clown6sh,it wouldalso be helptulto collectthe anemonethat the pair was occupying.Anemones arenot required for breeding most clown6sh. but are desirable if availablefor stnnutatingmore natural behavior and facilitating pair formation.Some speciesof clown6shprefer a particulartype of anemone.Mated pairs can usually be

PREPARED BY GADTC FINAL REPORT; CULTURE OF MARINE AQUARIUM FISHES

obtainedby placing together one large ciownfish i.e., a female!with a small clowafi.k immature or male ash!, or by placing together two imtrutturefish. However. these latter methodsmay take severalmonths to obtam maturepairs. Another alternative to collecting wild caughtclowafish pairs or trymg to pair up mdividuaifish. is to purchasepairs of tank- raisedclownfish. A U.S.compaay called The Breeder'sCoanectioa. a divisionof Aquatic Technologies,ofFers medium to large pairs of tank-raisedciowafish of the followiag species:Amphiprion bicinctus. A. clark'i. A. ephippium.A. frenasus.A. ocellaris.A. perrula, A. sandanacinos,and Premnas biaculeatus. Although this option may seem expensiveor impracticalfor a hatcheryin MicroaesiLit may be a good sourceof broodstockof clowafishspecies that are aot foundlocally aad not fiequeatiyimported. suchas A. bicinctusaad A. ephippium.

Broodstock tanks of about 140 to 160 liters should be adequatefor coaditioniag aad spawaiagmost species of cloem6sh.Usmg tanks that arelarger than required adds to the initial cost of constructionaad makesfor less ef5cieat use of resources.An exceptionis Amphiprionchrvsoprerus. which is reportedto requirea largertank for spawning Young, 1991A!. Broodstockof this clowttfish speciesshould probably be kept ia tanks of at least 400 liters. Larger or more active speciesof marineaquarium fish may also require tanks that are 400 liters or larger ia size to stimulatespvvaiag. Water depth may be as importantas volume for somespecies, aad deeper tanks may be desirable for speciesthat exhibit a largespawning ascent during courtship.

Strong hghtingis requiredto maintainhealthy anemones. Relatively strong lighting 5,000 to 50,000lux! is alsodesirable for bmodstockand grow-out cuhure, aad artificial lighting,preferably Rom fidl sl:ectnnnhght bulbs,may be usedto supplementnatural sunhghtif neededto reachthe desiredhght intensity.CoanaeaM hatcheries, such as C- Quest.aad formerlyAqualife, use translucent fibergiass roofs that allow about 50% of incommgsunlight to eaterthe hatchery facihty, which may result in hatcherylight levelsas high as 50,000lux oa a brightday ia the tropics.Spevaiag occurred ia the Blue-streak gobyand Banded coral shriznp at matiatutalight levelsof onlyabout 1,000 to 1,500lux. However,higher hght levelsmay have resulted in bettermaturation and more sequent spawamg.Natural sunlight has beenreported 'to aid in malataiaiaggood co!oratioala marineaquariaaa fish McVnrney,l985!. However,high light mteasitiesoccurrmg uader direct suahght e.g., about75,000 to over 100,000lux! will stimulaterapid growth of filamentou algaeaud add to the maintenance.Clownish hekl in tankswith excessive Ghzaentousalgae may develop more subdued or washedout coloration.Larvae of at least somespecies of marmefish are reportedto be sensitiveto stronghghtiag Wassiakand Brons. 1990!,aad rehN:ed light levels,000 to 5,000lux! shouldbe adequate.aad Ia somecases required for obtamiagoptimal resultsin larviculture.

Good nutrition is requiredfor broodstockto promotefrequent spawning, large spawn size, aad healthyeggs and larvae,Marme aquarium species have high nutritional requirementsbecause of thehigh frequency of spawning,aad a highquality balanced diet is needed ia addition to a diet of mesh or frozen and live seafoods to achieve optima! results D. Fletcher,per. comm.,1996!. However. a commercialdiet that meetsall the

41 PREPARED BY GADTC FINAL REPORT - CULTUS OF hL43UNE AQUARIA'M FISHES nutritionalrequirements for breedjngmarine aquarium species does not appearto be readily availableat this tune.

We low level of spawningin the ciownfish hejd at the GADTC may be due primarily to ineffectiveconditioning of broodstockas a result of de6cienciesin broodstocknutrition. There may have been insuf8cient quantities of vitamins or other nncronutrientsaad essentialfatty acids in the frozen diet fed to the broodstock.The 6'ozeadiet usedin this project was relatively low in iipids, anda higherlevel of fat may be requiredfor optimizing broodstock conditioaiag aadjuvenile growwut H. Ako, per. conan., 1996!. Live foods, especiallywhen entichedwith fatty acids and other autrieats, are an excellentfood for conditionmg 6sh for spmading.However, they were often not availableas a supplement for feedingthe broodstockdurmg this study. Also, the size and quantity of feedingsmay not have been adequatefor maturationand spawning. Due to restraintsin staf5ag and other resources,the broodstockwere usually fed only twice daily with the trozen diet or dry feed. Commercial clownish hatcheries typically provide four feedmgs daily to broodstock.including at leastone feedmgwith live food.

Broodstock should be fed four times daily, using a high quality 6'ozendiet and enriched live adult Artenria or other enrichedlive food. The &men diet shouldhave high levels of highly unsaturatedfatty acids HUFA! and particularly DHA docosahexaenoicacid!, carotenoids astaxanthm!,and vitamins especiallyvitannn C in a stabilizedform!. A good source of caroteaoid pigments are two species of hill Euphausia supeIba and E. pacigca!. Astaxaathiaand other caroteaoidpigments can also be obtainedcommerciaUy It I P~ I .1l» dd'' f I' I' yU pigtneatsia the diet. De6cienciesin HUFA, especiallyDHA, havebeen shown to result in lower stressresistance and lower egg quahty. Carotenoidsare important for enhancing coloration and may haveautritioaal beaefits as well, possiblyas antioxidaats.

Clownish juveailes may be moved Rom the larval tanks to nursery or grow-out tanks after about three weeks,but it is probablybetter to wait until day 30 beforemoving them Very young juveniles are more susceptibleto hmdhng stress,especially if they have any nutritional deficiencies,and lossescauld occur during the tnnsfer process.Relatively high levels of dietary DHA have beenshowa to reduce stress-relatedmortality in marine fish postlarvae Ako et al., 1994!. AdequateDHA Ieveis caa be supphedto larval clownfish through the use of commercialenrichment diets to enrich rotifers and Anemia with this HUFA. Oae feed manufacturerhas developeda diet for mahimahithat may give good resultsfor grow-outof marineaquarium fish andanother company has an Artemia flake diet enriched with HUFAs that has worked very well for improving broodstock performanceof freshwaterornammtal fish H. Ako, per. comm.,1996!. A frozenArtemia product, enrichedwith HUFAs, is also availablecomaierciaily and tnay be a good supplementor iagredientia broodstockand grow-out diets for marineaquarium species. Lipid levelsshould be above15'/o and perhaps as high as 25-30'/oto optimizejuvenile growthand broodstock egg production. The grow-outdiet shouldalso be foitified with vitamins especially with highlevels of vitaminC! andhigh levels of astaxanthin e,g., I.00 ppm or higher of astaxanthm!.

42 PREPARED BY GADTC FINAL REPORT - CULTURE OF MldtINE AQUAPJUlvi FISHES

Grow~ut of juvenile clown6sh was not optinm~M in this study, although some improvementwas made later in the project. Market size fish should be producedwithm approximately3 to 4 months of grow-out under good conditions. Slow growth in some batchesof clownfish probablyresulted 6'om inadequatefeeding levels and nutrition. Fish in grow-out were only fed twice eachday, whereasin most commercialhatcheries the grow-out 6sh are usually fed four times per day. Hm method of feedingthe frozen diet was alsonot very ef5cient Le., largerpieces of food o6en fell to the bottom andwere not eatenby the 6sh!, prmiarily due to the lack of proper equigenentto grmd the food to a smail uniform size. As a result, the amount of food consinned at each feeding was probablyless than required for good growth. The nutritional value of the grow-out diet was not optimized in this study and shouMbe improved. However, someimprovements were madein feedmgthe juvenilesduring the courseof the study, and better growth was obtained.Fish should be size-sortedat Sequentintervals during the grow-out period to preventinhibition of growth in the smallerfish. Deformedfish shouldalso be culled out as soonas possibleduring the growwut period ie., as soon as the fish are large enoughfor deformitiesto be detected!. Early cuomgwill decreaseoverall costs of production and providemore spaceand fastergrowth in the remalaingfish.

The higherincidence of deformitiesin juvenileclownfish Sum a coupleof the latter spawnsreared may have beendue to a nutritionaldeficiency in the broodstockdiet. because ~~nins were oAen omitted Rom the Glen broodstock diet during the last severalmonths of the grant.Fletcher and Wihon 996! foundthat vitiumnC de6ciencies in the diet of AmphiprioecIantii broodstockresulted m significantincreases 6/o to 83/o! in the inadence and severityof certamdeformities. In addition, the 24 hour post- hatch larval survival was 10/o to l 5/o lower in these fish.

However,deformities may have also occurreddue to problemsin 1arvalrearmg. Instant OceanHatcheries found that individualtanks of fish frequentlyhad 50/o to 80'/a saiable fish and 80'/oto 98'/o survival,while other tankshad 10/a to 20'/o salable Hoff In Prep.!. However,since both types of resultso8en camefrom fish &om the samespawns. it appearsthat the problemoccurred due to rearmgconditions in the in&&ual tanks probablya combinationof waterquabty and feeding/nutrition problems!, Enrichment or booster diets that contam vitamins can be used to increase the vitamin level m rotifers and Artemia fed to the larvae, and this may also aid m reducing deformitiesand increasmg overallhealth of the hrvae. The time around metamorphosisappears to be a particularly sensitivetime, where suboptimal conditions can lead to increasedincidence of deformities F. Hoff per. comm..1996!. The low incidenceof deformitiesin fish fiom mostof the spawnscultured at the GADTC suggeststhat problemsof deformitiescan be kept at a very low level if time and resourcescan be put into optimizing diets and rearing conditions.

Colorationwas good during the first halfof the study,when 8'esh shrimp was available for use in the clownfish diet. The shrimpwas believedto provide carotenoidpigments in the diet.However. shrimp was not availablehter in the studyand this probably contributed to

43 PREPARED BY GADTC FINAL REPORT,- CULTURE OF IvVdWW AQUARKM FISHES the reductionin bright coloration observedtowards the exl of the project. This coloration problemcan easily be correctedif sufficientquantities of carotenoidpigments are put back mto the diet.

The Zebra goby pair spawned regularly throughout the study, indicating that the broodstockdiet and maintenancewere adequatefor promotmgspawning. However. there were a coupleperiods of time when the interval betweenspawns was extendedto about twice the normal interval. This may have been due to inadequatebroodstock nutrition during certainperiods, or it may have beena naturalperiod of reproductiveinactivity,

The Blue-streakgoby pair spawnedregularly before beingmoved to another smallertank. Thisdishebance probably disrupted their spawningbehavior. One of the fish alsobecame noticeablythinner aRer the move,and may not havebeen gettmg enough food to maintain it in spawningcondition. It is aLsopossible that spawnmgmay be seasonalin this species, althoughmost reef fish appearto have extendedor year around spawning,especially in locationsnear the equator.

A varietyof shapesand sizes of tankscan be used for larvalrearing, including aquariums. However,round fiberglassor plastictanks with a depthof thirty inchesor more are emdard for rearmglarval marinefish, and probablyare the most e%~ design.The sides of the tank should be a dark color to provide a better visual contrast of food organismsagainst the background,and white bottoms allow for easierobservation of the larvaeby the larval culturist. Tanks of 300 liters or larger are preferableto smallertanks for commercial culture siteltions. th ~ ' f 'I Wwl ~ ' I ' u GADTC ~ has beensucoesfidly usedas a 5rst food for larval rearmgof Amphiprion melanous and the early stagesof Steopus hispidus. The ss-rotiferis smallerthan the s-strainof rotifer andshould have better potential for rearingsmall larvae of somenMu~ aquariumfish. However,larval rearing of the Blue-streakgoby and Zebra gaby remains unsuccess%4 due to the lack of a scrutable-sizedlive food during the Grstfew days of feeding.The larvaeof both spam of gobyare appareadytoo smallto eat ss-typerotifers at first-feedmg,and completemortality always occurred after a few days,probably due to starvation.The larvaeof manyother species of marineaquaxiaun fish have been reported to betoo smallto eatrotifers at first-feeding,and there have aLso been reports of the larvaeof somespecies e.g.,Pomacanrhus sp.! refusing to eatrotifers even though the larvaeare largeenough to eat them Young, 1994Aand 19948!. Sievingthe smallestrotifers out of the GADTC cultureswas not attempted.but maygive better results than using the entiresize range of the rotifer cultures. Seaurchin larvaewere not obtamedon a dependablebasis, and adequatequantities were not availableto accuratelyevaluate its potentialfor rearinggoby larvaeand other small fishlarvae. It is not knownwhether problems in spawningthe sea urchins later m thestudy was due to nutritional deficienciesin the seaurchins heM at the GADTC, or if spawningin this speciesis seasonal.Also. the seaurchm larvae grow fairly rapidlyand need to be fed soonafter hatching Figures 12 and I 3!.Fertilized sea urchin eggs may also have potential

PREPAREDBY GADTC x'21'.; ' 34 J7Z:;;[! Ale'Lpc'."i hj- Sd;I ".. Uhim I I ' .'.'8!f I'>> PQ Frt~, fl, ! il,". >'S >It"'; ter.~i.;xi 'J. ju'L '.':l 'gill >~- i rvicr': .'i

i'." i.4,: L'~ i' .. I»[C'.g. g'.J<' ~gg '~.i lz' ~~.'qL' g; 'j b s~]74LCPLv. ' g'jj'~'!'pi'.i'. gl I!Ill II -. i I Q;~, [...g FINAL REPORT- CPLTUPX OF 1%443UNKAQUARIlJM FISHES

asa livefood. but some method of suspendingthe eggs m thewater column is neededthat wiH not damagethe larvae. Similarproblems were encountered in.producing oyster larvae. Induced spawning was onlysuccessful ontwo occasions. although the procedure was attempted over 20 times. Attemptsto dissectout the gonads and manually nnx the eggs and sperm werc also uasuccessfuLbecause there was very little eggsor spermia thegonads. mdicating that the oysterswere not ripe for spawning. The species of oysters fouad on Guam are reported tohave some spawning activity throughout the year, suggesting thata nutritionalproblem preventedthem &om being in spawamgcondition. Oysterand sea urchin larvae may still have good potential as a livefood for marine aquariumGsh larvae. but methods aced to be developed forconditioning andspawning the adultsin captivity.Braley 982 aad1984! provides information on thereproductive biologyof theseoysters. The major peak in spavmiagactivity in the largeroyster Crassosrreaech inara. occurred in Marchand the dcvclopmeat of another was observed ia lateNovember. The smaller species. Saccosnea cucullata, was found to havea lowlevel of continuousreproduction with three main peaks throughout the year, in November- December,March-April, aad late June. The dif5cuhy in spawningthese oysters. especiaHy C.echinata, may be due to naturalseasoaal reproductive periodicity. Oyster trocophores arereported to bea gooderst food for successfblculture of somespecies with small larvae e.g., Yellowtail snapper!, but the larvae of someother species will not eat them J. Holt,pcr, comm.. 1996!. Alternate live foods should be examined for culturing species withvery smaH 6rst-feeding larvae. Other possible live foods include cultured copepods. aematodes, and ciTiates. Fecundityinthe Banded coral sb~np did not declme with time m captivity,indicating that broodstockmaintenance aad nutrition were adequatefor spawnproduction. However, improvementscould be made in thebroodstock diet aad feeding to mamtainmaximum outputof quaiitylarvae. The spawning data shows that thc Baaded coral shrimp are able to spawnevery two to threeweeks throughout the year. A fewgapa ia thespawning recordfor two of the pairs is more likely due to unregertedorunobserved hatches rather thanactual disruptioas in theirspawning rhythms. Althoughonly a limitednumber of rearmgtrials were attempted, larval rearmg of the Bandedcoral shrimp improved during the study, and in themost successful trial some larvaeappeared healthy and probably would have survived much longer if the tank had aot becomecontatnmated withhydroids and jellyfish. The appearance of large numbers of hydroidsand smaH jeHy6sh medusae inthe larval Banded coral shrimp tanks shortly before completemortahty ofthe larvae strongly suggests that the hydroids and mcdusae were the causeof the mortality.possibly due to stingmgthe larvae.However. the larvae were biggerthan the medusae andno larvae were observed to be captured bythem. It is unlikelythat the hydroids and medusae fouled the water or ate aH the Anemia. since the tanks were oa a continuousflow-through of seawaterat the time and there was aa abundanceof Arremiain the tank. The hydroidsand jellyfish mcdusac have occasionally

46 PREPARED BY GADTC FINAL REPORT- CULTIC& OF Iv&QUNEAQUARIUM FISHES

beenobserved in Macmbnrcht'umlarval rearing tanks at the GADTC, and may come in with the seawater 6'om the saltwater weHs. Contannnation of the larval tanks with hydroidsand jeHyfishwas successfuHyprevented in the last couple of larval rearing trials by filtering the water with a five micron Ster bag. However. only Artemia was usedas the larval food in thesetrials to avoid possiblecontammation &em the rotifer cultures.and the 1arvaislirimp experiencedrelatively large mortalities m the first two weeks and the trial was discontmued. Better survival of Banded coral shrimp 1arvaecarly in trials where rotiferswere fed suggeststhat the rotiferswere morenutritious than Aeemia naupiii for the larvaeor that the larvaewere heing dUKcultycapturmg and consiuningenough of the Arternia.

No specialenvironmental cues are requiredto initiate metamorphosisin Bandedcoral shrimp D. Jones.per. comm., 1995!. However, the larvalperiod is verylong compared to most fish or shrimp 1arvae,and laboratoriesthat have successfbHyreated Bandedcoral shrittip larvae through to the juvenile stage report me3morphosisoccurring after more than 100 days D. Fletcher. per. coast. 1996; L. CresweH.per. comm., 1996!. The extremelylong larval period of the Bandedcoral shrmapmakes the commerciahzatioaof this speciesmore dif5cuit comparedto other more vahable aquarnnnshrimp with shorter larval periods. In additionto the addedtime and expenseof raisingspecies with long 1arval periods, there is a higher probabihtyof problemsdeveloping before the 1arvaereach the juvemlestage L CresweH,per. comm., 1996!. The larvaealso appearto be very sensitive to nutritional de6ciencies D. Fletcher,per. comm., 1994!.

Objective 5. There appearsto be a growing trend towards tigi~ restrictions on the harvest and import/export of marmetropical 6sh and invertebrates.This trend should ultimately work to the advantageof the coaxial hatcheries,with the reduced availabahtyof wild~ught fish maitingthe tank-raised6sh more competit5rein the market. It is quitepossible that at sometime in the batture.the harvestand sale of wHdwaughtreef fishand invertebrates will be completelybannoi for thosespecies that arebeing cultured in captivity. Pyle l993! providesadditional information an the statusof the marine aquariumindustry in the Paci6cregion.

ObIective6. Maricuiturecertification does not appearto posea problemfor thegrowth of the industry. Governmentceration of approvedhatchery facilities, with periodic inspectionscould insurethat marineaquarium species are captive-bred.

Objective7. A clown6shhatchery and growwut facility can be profitabieif a reliable productionof marketablejuveniles can be achievedand all marketablefish produced can beMid at a ininimumprice of $2.25each. The busmesscou1d be more prohtable if several speciesof clovra6shwere raised,especiaHy 1ess conznon species. to achievemore diversityand a slightlyhigher average price. Higherprices for marketsize clownfish are possible,and this wouldresult in a higher1evel of profitabihty.Reef Propagations is obtainingbetween $3,25 and $4.00 per fish J. Lichtenbert.per. comm.. 1996!, and Desert Fisheriesis obtaining$6.00 per 6sh L Nelson.per. comm., 1996!. A hatcheryfor tnaiitie aquariumfish would alsobe morepro6table if someproduction of otherhigher vaiue

PREPARED BY GADTC FINAL REPORT; CULTURE OF M 4Ch24EAQU/JUUM FISHES species e.g., Pseudochromissp. or the Comet!was addedto supplementclownish productionand achievea higheraverage price for Gshproduced perhaps two or three activ~pairs of eachof the otherspecies!, Inactive broodstocL should be replaced quic]tly with activespawning clovm6sh or othergecies. Additionallow cost, low maintenance holdingtanks could be addedto keepnew broodstock in reserve.Examples of speciesthat could be cultured in a commercialhatchery in Micronesia and their estimatedfarm gate value are listed below:

MicronesianSpecies Farm Valse ' Amphiprion bicincrus $3.75 A. chrysopterus $4.00 A. clarkii $2.25 A. ephippiuIrt $3.75 A. melanopus $2.25 A. perideruion $2.50 Pseudochrornisporphyries $10.00

Imported Species Farm Valse * Arnphiprionoceklaris $2.50 A. frenarus $2.50 Calloplesiopsaldvelis $10.00 P~nas biacrdearus $3.50 Pseudochmmis dutoiti $15.00 P. flavhertex $10.00 P. fridnumi $10.00 Hymertocempicta $9.00 Lysmatadebeh'us $10.00 Lysmata grabtuaai $7.00 ' Estimatedor potettial vahteto producerbased upon informationobtained Qom commercialproducers or wholesale price lists. Wbea prices varied between producers. the lowestprice normally obtained was used for theseestimttes, Someof thehigher-priced species are harder to produce e,gAmphiprion chrvsopterus. CaBoplesiopsalrivelis, anal Lvsmcrm debelius! or canonly be marketed in smail quantities e.g.,Pseudochromis sp. andHynumocera picta!. However.the productionof a smail quantityof thesehigher-g6ai spom in additionto themore easily cultured and lower- pricedspecies can improve the economic return and the larger product diversny weal facilitatemarketmg efforts.

PREPARED BY GADTC ANAL REPORT-, CULTL RE OF M/dUNK AQUARIUM FISHES

RECOACi%%PG!ATIONSFOR CULTURE OF MAR1NEAQUAIUUM SPECIES:

Co0ectaaturaHy mated pairs from the wild when possible. For clownish broodstock.try to collect the aaemoaethe pair was occupying or at leastanother anemone of the samespecies. lf mated pairs of cloddish cannot be collected.place together one large clo~~ i.e.. a female!with a smallclowa6sh immature or maleGsh! less than 4 cmin length. UM broodstocktaaks aquariums!of 120 to 160 liters for clo~~ broodstock.For AIrrphiprionchrvsopr urus. use a tank of at least400 liters. Use strong hghting ie.. light intensity of 15,000to 50,000 hx! for broodstock and growwut, Usenatural sunlight or Adl-spearumbulbs ia arti6cial lightiag. Use lower light levels i.e.. light mteasity< 5,XO hx! for larval culture. Feed broodstockfour times daily, using a high quality Sozea diet and enrichedlive adult Arrerrria or other emiched live food. A high quahty dry diet formulated for acme 6sh culturemay be substitutedfor one feeding,if available. Frozen diets for broodstock and growwut should have lipid levels above 15 o aud high levels of HUFAs particularly DHA!, caroteaoids primImly asta:samhitaat 100 ppm or higher!, aadvitamins especiallyvitamin C aadvitrunia E!. Use round tanks with depth of thirty inchesor more for larval rearing. The tanks should have dark sides aad a white bottom. Feedlarval clown6shenriched rotifers for the Brst tea days, aad Artemia aauplii 6om day 7 to day 15. Enriched 24-hour Artemia and small particle size arti6cial diets should be fed beginning at about day 15. 1%is feeding schedulemay need to be adjustedbased upon growth rate of the larvae. Use live foods e.g., rotifers aad Arternia! that are enrichedwith HUFAs aad vitamms wheneverpossible in larval culture. Use microalgaeduring the rotifer feedmgstage of larval culture. Speciesthat caa be used inchde Chaesocems gnrciNs, Isochrysis sp., Nanrrochlorepsis omiaca. Rhodomormssp. and Tetnrsebsissp. Use daily water exchangeor continuousSow-through of seawaterafter the larvaeare feedingon Arremia. Move clovIm5shjuveniles Seta larval tanks to nurseryor growwut tanks at 25 to 30 days. Move a few Bahand watch for htadling stressproblems before moving all the Gsh ia a tank. Stock larval tanksat ao more than 5 Gshper hter. Stock growmut tanksat ao more than one 6shper liter. Feed 6sh four times per day in grow-out. with two feedingsof a high quality frozen diet and two feedingsof a high quahty dty diet Qake or crtnable! formulated for marme Ash. Youngjuvenile clownish should receive a supplementof live enrichedArterrria for >he Bra two to threeweeks after they are transferredto the nurseryor grow-out tanks. Sortjuveniles for sizeat 6 equeatmtervals during growwut. Examinejuveniles at an early agefor deformitiesand continue checking them throughoutthe grow-out period. This can be danewhen size-sortingthe Ash.

PREPARED BY GADTC ~AL REPORT - CULTURE OF MAJEUREAQUARH;M FISHES I

Deformedfish should be culled out assoon as possible and not offeredfor sale except possiblyas feederfish!. ~ Off-color 6sh should be fed diets with higher levelsof carotenoidsfor 4 to 6 weeks beforethey reachnMttket size to 4 mm! to improve coloration. ~ Keep excessivefilamentous algae growth out of all rearingtanks. ~ Use hve foods e.g., rotifers. Artemia. nematodes!enriched with HUFAs, vitammsand carotenoidpignMnts for larval culture of marineaquarium ~imp.

CON CI.USIONS.'

Advances have recently been made in .the cttlture of nmrine aquarium fish and invertebrates,as reQectedin the literature and the increasingdiversity of speciesunder commercialcultivatiojL Although somemajor tmmneannum fish hatcherieshave closed operations in the last few years, the remaining coamercial and researchfacilities are improving their production capabilitiesand econotnic potentiaL lncreaed diversity af speciesand production of more highly-valuedspecies are positivetrends occurring in this 19dustry.

Severalmarine aquarIlnn species occurrmg in Micfonenahave been ldentlfied as priIQe candidatesfor aquacultureproduction. Some of these,such as the clownish species, some of the gobies,the Comet,«nd the Magentadott Itback.have been successhdly cultured in otherparts of the world. Othervahusble Micronesian species may prove to be conducive to captiverearing with ac}ditI>nalresearch.

The Cinnamonclownfish hmphiprion eeranopus, was success53lly cultured in this project anddemonstrates the feasibly of marmeaquarium fish culturein h5cmnesia.Spevning was achievedfor two speciesof goby, althoughthe tmy larvaewere not successtuily reared.A hvefood that is sigai5cantlysmaller than the ss-strainof rotiferwill be required in largequantihes to rearthese species and other v~dmAe marine aquariMm fish Mt have very small first-feedinglarvae. Problems in obtainingmated pairs of clownfishcan be solvedthrough increasoi events at coQectingmated pairs Som the wild or by collecting smalljuveniles and pairing them with otherjuveniles or adultfish althoughthese latter methods will take more tine!.

Betterquality premiered diets and enriched live feeds.as well asimproved feeding methods can be used to properlycondition the broodstockpairs to achievebetter spawning performance.as well asimproved larval quality and survival through first-feedmg stages. New live feedsfor specieswith very smallfirst-feeling larvae are currentlybeing developedby a privatecompany in theUnited Kingdom. incretsing the potential for larval cultureof manyreef fish D. FletChe.per. comm..l996!.

Goodgrowth was obtained with onebatch o fclown6sh juveniles. and marketsize ta 4 an! fish can be obtaineRafter three to four monthsif attentionis givento optimizing nutrition and feeding.Marketability, ia termsof colorationand appearance

CQ PREPARED BY GADTC FPZAE. REPORT - CULTURE OF!VbVtZW AQU/GGUI41 FISHES

deformtties!was high with most batche- ~f clownish juveniles reared in the study, and high quality Qshcan againbe obtaine; by optimiziugnutrition during all phasesof production.

Regular spawning of captive broodstock of the Banded coral shrimp occurred during much of the study and consistentproduction of newly hatchedlarvae should not be a problem for their culture. However, larval rearing of this speciesis a long and dif6cult process,and the potential for their eventualconmMscial production is poor, Other species of marine aquariumsshrimp, such as Lvsmaradebelius and Hymenoeerapicta are much morepromismg candidates for conunmM culture,

Regulatoryconstraittts for the import/exportof marineaquarrutn species will increasein near tuture. but this trend towards restrL~ the coDectionof wildwaught animalsfavor should work in the favor of captive breoMg progr3ms. Mariculture certification of cultured aquas Qsh can be done without much dif5cuhy through certification and aenitoring of government-approvedculture facilities.

Competition in the aquariummarket with cheaperwild-caught Qshand shrimp presently makescommercusl culture of ma3~ aquaziutn6sh and shrinspmore diBicuit economically. However,the comttMsIeialculture of variousmarme oraamesttal species could be pro6table in Micronesia if the businessis set up properly and well managecLThe future for commercialmarine aquarusm ash culturelooks evenbrighter, as more speciescome under cultureand the regulatory climaterestricts the collection of wild~ught reef Gsh.

LlTERATURE1ss> CHX9:

Baker,D.E., Jr., 1993.The Onslaughtof Tropical Marine Auparium Fish Harvesting:Are They Really Rapingthe Reefs?Freshwater and Marine Aquarium.VoL 160!:168-173,

Braky, R.D., 1982. ReproductivePeriodicity in the indigenousOyster Saccosrrea cundlaazin SasaBay, Apra Harbor, GuIsm. Marine Biology, 69:165-173.

Braley, R.D., 1984. Mariculture Potetttial of Introduced OystersSaccostrea cucullara tubercuIaraand Crcrssostreuechinara, and a Histological Study of Reproductionof C, echinata.

Broas,R., 1995. Red SeaFish pHarm. The Breeder'sRegistry, 3 l!;1-6.

Brons. R.. 1996. 1bqmduction and Captive Breeding of Two Red Sea Dottybacks: Pseudochremisfridmani and P. flaviverter, Freshwaterand Marine Aquarium. 19!:48- 62.

Burgess.W,E.. Axehod.H.R. andR.E. Hunziker.1990. Dr. Burgess'sAtlas of Maine AquariumFishes. T.F.H. Publications.Inc.. Neptune.'.4.J., 768 pp.

5l PREPARED BY GADTC FINAL REPORT - CULTURE OF M,4JUXK AQUARIUM FISHES

Damlowicz,B,S. and C.L. Brown. 1992.Rearing Methods for Two DamseliishSpecies: Dascvllusalbisella Gill! and D. an>anus L.!. Aquaculture,106:141-149.

Fletcher. D.J. and M. Wilson. 1996, Vitamin C Requirementsof the Anemone Fish, AmphiprionchrhL Page 128 In: Book of Abstracts- World Aquacuhme '96, The 1996 AnnualMeetmg of the World AquacultureSociety, Bangkok. Thailand, 498 pp.

Gutierret, S., 1996. From a Reefs Point of View. OrnamentalFish Farming: How and Why, Freshwaterm} Marine Aquarium, 19!.203-214.

Hoof,F., In Preparation.Conditioning, Spawning and RearmgMarine Fish. Florida Aqua Farms,Dade City, FL.

Liao, I.C., M.-S. Su and S,-L, Chang,1995. A Review of the Nurseryand Growout Techniquesof High-ValueMarine Finfishes I Taiwan. In K.L. Mam and C. Rosenfekl Kds.! Cultureof High-ValueMarine Fishes in Asiaand the UnitedStates. The Oceanic Institute,Honolulu. HI, pp.121-137.

McLaraey,W., 1985.Pioneers in SaltwaterAquaria@a Fish: Part L AquacultureMagazme, 1 1!:3841.

Myers, ILF. 1991.Micronesmn Reef Fishes:A PtmcticalGuide to the Identificationof the inshoreMarine Fishes of the Tropical Cetttraland WesternPaciiic. Coral Graphics.Guam. Second Edition.

Pyle,R., 1993.Marine Aquariann Fish. Pages 135-176 In: AndrewWright and Lance Hi11 Editors! NearshoreMltrine Resourcesof the SouthPacdlc.

Schiller,G., 1990.Spearing Behavior of the DecoratedFire6sh V'eernreleotris decora!. SeaScope,7:4. FaIL

Tamaru. C.S. and C. Carllrom-TriclL 1995. Striped Mullet Mugil cephalusTaiwan Strain!Hatchery Pmhlcthaa and Growout Demonstration. Final Report Submixted to the PacificAquaculture Development Program, Jammed' 22. 1995.Htpvaii C's Aquaculture Consultant Services. KLitua, |II.

Taint, C.SW.J. Fitzi3eadd. and V.S. Sato. 1993. Hatchery manualfor the artificial propagationof striate mullet Mugil cephaius!.C. Carlstrom-Trick Ed.!, Guam AquacultureDevelopment and Trains Center,165 pp.

Torben.J., 1992.Commercial Breeding of Anemoneashes.SeaScope, 9;1-4, Winter.

Walker.S.D.. 1991.Reproducing the AtlanticSpadefish at theTulsa Zoo. SeaScope,5;1- 2, Winter.

52 PREPARED BY GADTC FINAL REPORT - Cia'LMRE OF MISTUNE AQU&GLM FISHES

Wassink.H. andR. Brons. 1990.A SuccessfulCultivation of the CometCalloplesiops alrivelis SteiadaMer, l903!. SeaScope,7:1-3, Sprmg.

Wi9rersoa.J., 1996. Cghest Hatchery: hnovations In CaptiveCmental Marine Fish Culturing. Freshwaterand Marine Aquarium, 19!:123-13 8.

Youag, F,A.. 1991A. Breediag the Orange-Fia Anetaone6sh.Marine Fish Monthly, 6!:46.

Young, F.A., 1994. Food Preferencesin Tropical Marme Fish Larvae. Aquaculture Magazine,Vol. 20 l!:57-59.

PERSONAL COMMJNICATIONS:

Addison, WiHiam.C-Quest. Peurto Rico, Phone:S09-845-3909 Ako, Harry, Universityof Hawaii, Honolulu, HI, Phone:S08-956-2012 Browa, Chris. Universityof Hawaii, HBAB, HI, Phol: S08-236-7445 Brown, Stanjey.Tbe Breeder'sRegistry, Sacrmaeato, CA. Phone:916-487-3752 Cresweli,Leroy, Harbor BranchOceanogmphic Institute, Fort Pierce,FL. Phone: 407-465-2400 Davis,Gerry. Department of Aquaticaad VItlife Resources,Mangilao, GU. Phoae: 671-735-3958 Fletcher,David. University of WaJes,Bangor; UK. Phone:44 1248351151 Kxt. 2900! Fakes,Tom. Aquarmm System, Mentor, OH. Phone:800-822-1100 Holt, Joan.University of Texashhtrme Science Institute, Port Aransas, TX. Phoae: 512-749%716 Hoff Frank.AqmLculture Supply, Dade City, FL. Phone:904-567-8540 Kraul. Syd.Paci6c Phmktoaics, IGtiluL HI. Phone;808-262-5134 Lichtenbert.Joe. Reef Propayions, Ho5aaa Estates.IL. Phoae:708-885-733811 Moe, Martha GreenTurtle Publications,Plantation. FL. Phone:954-384-8382 Myers. Robert.Coral Graplnca,Bamgada, GU. Phone:671-734-2590 Nelson.Linda. DesertFisheries. Gmee

53 PREPARED BY GADTC ~AL REPORT - CULTS'RE OF ~MVUNE AQUARKiM FISHES < ~

APPKNDIX L FROZKN DIET FOR l4GQUKE AQUARR9f FISH

GEL MX RECIPE MODIFIED %ALKI AQUARIA'M RECIPE!

1.00kg ash marineash preferred: smelt. reef 6sh.etc...! ' 1.00kg squid ~ 0.50 kg trout chow 0.35 kg shrimp ~~' 0.85 kg vegetables 50 g yeast 3.8 g Vitamin C

3.8 kg subtotal 3.8 L water 7.6 kg TOTAL Before Gelatin!

760 g Knox Gelatm0%!

' Other seafood items can be substituted for part of the Bah or squid, includmg the foEowing:clams, oysters, scallops, marine bloodworms, crab meat, and fish eggs. '~ Combinationof two or more of the followmg: zuccim,spmach, carrots, green peas, or romaine lettuce. '~~ Usewhole, shell-on shrimp. Usmg only shrimp heads or a toohigh of a percentageof slirimpwill causethe am not to gelwith 10%knox gelatm.

DIRECTIONS: Measureout 1750ml of waterto be usedm the grmdiagprocess. 'He remiunderof the water050 ml! canbe setto heatfor dissolvingthe gelatin.Chop aE ingredients into smallpieces and grind in theblender with suEcientwater. Add ingredients in aboutfour equalparts with theaddlition of watereach time and using a spatulato pushsolid pieces toward the blades. Putgelatin in a largemixing bowl. add the boilmg water 050 ml!,and mix with the electricmixer. scrapmg the sides and bottom oAetL After throughly dissolved. allow the mixoueto coolto lukewarm,mixing occasionaEy to hasten cooling and keep gelatin Qotn lumping.When cooled, add ground mgredients to dissolved gelatin and mix. Next add the VitammC andthe yeast but onlywhen the ingredientsare lukewarm or cooler, and continuemixing. Pourthis mixture into 4 to 5 traysand allow to setin thefreezer for 12to 15minutes. Removetrays before the mix freezes and cut mto squares. Package the squares in the zip tock bagsand storein the teezcr.

PREPARED BY GADTC FINAL REPORT,- CULTS'~ OF h4DUNK AQUARlUM FISHES

APPENDIX K Aquarium HobbyLiterature on the Cultureof Marine Aquarium Fishand Shrimp

Addison.81994A, Spawningand Reariag the BlackcapBasslet. Re Breeder'sRegistry, 2{3!:1-4.

Addisoa, B.. 1994B. Spawningaad Rearmgthe Blackcap Basslet Gramma melacara!. SeaScope,11;1-3. FalL

Algosaibi.F., 1953, Spearing MandarinFish. Freshwateraad Marine AquarnaL 6{S!:8.

Anon., 1976.On Reariagthe Fry of MImae Tropicals.Tropical Fish Hobbyist,251!:32- 34.

Anon., 1984A. Conditioaing and Spawning of Marine Fishes, Part 1. Adult Pairs. SeaScope,1:1-4, Spring.

Anon.. 1984B.Larval Rearing,Part IQ: Marine Fish Culture. SeaScope,1:1-3. FalL

Amdt, D., 1989.Crossbreeding CovrnBsh. Freshwater and Marine AquariutL 12{8!:8-12.

Bauer,S. and J. Bauer, 1974.Spawjsing the CatalinaGoby. Octopus, 1 8!:6-11.

Bauer,J.A. aad G. KIaij, 1974.Pigmy AngelsSpawn. Octopus, 1!:7-18.

Behmga, S.D., 1993. Clowa5sh Broodstock: Selection and Pairing. The Breeder's Registry,1!:1<.

Boyd, D., 1956. Spawningthe Neon Goby Elacahnwsocernops. The Aquarium,25:390- 396.

Broas,R., 1995.Red Sea Fish pHarm. The Breeder's Registry, 3 l!:1-6.

Brosseau,R., 1991. Yet Another Clot~ Breeder.Freshvater and Marine Aquarium, 14!:112-120.

Brown, S.D., 1994. ProSmg Ceemgryge.The Breeder'sRegistry, 2!:1-6.

Brown, S.D., 1996.Gobies. The Breeder'sRegistry, 4!:1-5.

Cobb,R.V.. 1975.Spawamg Cloudy Damsels.Marine Aquarist, 6!:4649.

Co1emaa,R.K.. 1966,Spawaing Red Cardinals. Salt Water Aquarium, 2!:144-148,

55 PREPARED BY GADTC FINAL REPORT - CULM' OF MVUNE AQUAIGUTvf FISHES

Cooper, E., 1991. An Introduction to Rearing Larval h8ariae Fishes. Freshwaterand MarineAquarium. 3 4!:120-128.

De&mnardo.J.P., 1976. Spawnmgand Rearingthe Blue DeviL Tropical Fish Hobbyist. August,pp. 90-98.

DiMarco, R., 1994,Clownfish Breeding.The Breeder'sRegistry, 2!.1%.

Emmens.C.W., 1988,Reproduction in MarineFishes. Chapter 10 in MarineFishes and Invertebratesin Your Own Home. TFH Pubhcations,inc., NeptuneCity, N.J., 192pp.

Etherinyon,W1972. MatingBehrvior of theFiji DamseLMarme Aquarist, 3!:32-34,

Frakcs.T., 1992,Notes on BreedingQownfisk SeaScope,9:2, Winter.

Goldstein.R.J., 1989. Breedmg Marine Ciowtifish. Aquariaun Fish Magazine, 1!:18-23.

Goldstein,R.J., 1990.Tomato Clovm6sh,Bred and Raise m the Home. Aquarium Fish Magazme,3!:56-59.

Gutierrez, S., 1995. From a ReeksPoint of View. C-Quest OrnamentalFish Farming: Howand Why. Freslnvater and Marine Aquariinn, 181!:72-76.

Gutierrex,S., 1996. From a Reefs Point of View. OrnamentalFish Farming: How and Why. Freshwaterand Marine Aquarium, 19 l!:203-214.

HofF,F., In Preparation.Conditioning, Spawning and Reiring Marine Fish. Florida Aqua Farms,Dade City, FL

HoE F. andT. Snell, 1987.Plankton Culture h4snuaL Florida Aqua Farms.Dade City, FL. Hough,D., 1993.Breedmg Marhe Fishand the GreenRevolution. Freshwater and MarineAqetrium, 16!:152-158.

Karanikas.J., 1989.The Spawningof the FlameAngel Centmpyge joricuhcs. SeaScope. 7:1-2. Spring. Kloth.T.C., 1979. Breedmg and Raising Tropical Marine Fish: Part One. Freshwater and MarineAquarium. 2!:44-76. Kloth.T.C., 1979. Brexbug and Raising Tropical Marine Fish: Part Two. Freshwater and MarineAquarium. Vol 2!:22-78.

56 PREPARED BY GADTC FINAL REPORT - CULTURE OF h&dUNE AQUARIUM HSHES ~ ~

Kloth. T.C.. 1979. Breedingand Raising Tropical Marine Fish: Part Three. Freshwater and Marine Aquaritmi,Vol 2!:40-69.

Kuiter. R.H., 1988. Notes on Hatching and Raisiag of Seadragoas.SeaScope, 5:1-3, Sumtner.

KrauL S. and A. Nelson. 1986.The Life Cycle of the HarlequinShrmq. Freshwaterand Marine Aquarium,9 9!:28-31.

Laxnm,D.R., 1984. Spavmiag of theCoral Beauty AngeUish. SeaScope, 1:1-3, Summer.

Liader, R., 1986.Some Notes oa BreedingClownish. Freshwaterand Marine Aquarium, 9!:45-51.

Lobel, P.S., 1974.Another Pygmy Angel Spawns.Octopus, 1!:24.

Label. P.S., 1977.Spawnmg Behavior of ReefFishes. The Marine Aquarist, 8!:6-16.

Loch, M.G. 1995.Report of SpmvnmgGreen Chromism Captivity: Chromis caeruleus. Marine FishMonthly, 10 9!:13-14.

McLarney,W., 1985.Pioneers in Sdbvater AquariumFish: Part L AquacultureMagazine. 1 l!:38<1,

McLaraey, W., 1986. Pioneersin Saltwater%barium Fish: Part H. Aquacuhure Magazine,12!.31-33.

MtIs, D., 1987.Breedmg Marme Fishes. Pages 54-55 m TheTetra Encyclopedia of The Marine Aquarium,Tetra Press,Morris Plains.NJ, 208 pp.

Moe, M.A., Jr., 1973A. Breediag the Clovm6sb, Amphiprion ocelians. Salt Water Aquarium,9!:3-14.

Moe, M.A., Jr., 1973B. Commercial Breeder DiscussesBreediag and Rearing of Clownfish.%%trine Hobbyist News. 1!:1-3.

Moe,M.A., Jr., 1975,Propagating the AtlanticNeon Goby. The Marine Aquarist. 6!.4- 10.

Moe,M.A., Jr., 1976.Reariag Atlantic Angelfish. Marme Aquarist. 7!:17-26.

Moe,M.A., Jr., 1977.Inside the Egg of anAngelfish. Marme Aquarist. 8!:5-13.

Moe. M.A.. Jr., 1978. The Private Oceans. Sea Frontiers. 24!;240-247,

57 PREPARED B Y GADTC FINAL REPORT - CULTURE OF I<48UNE AQUARIUM FISHES

Moe.M.A., Jr,, 1981.Jus! Passing Through...Aquaiife Research Corporation. Freshwater and Marine Aquariung4 9!:26-79. Moe.M.A., Jr.. 1989ABreeding the Clownish. Amphipn'on ocellaris, Part L Freshwater and Marine Aquarium. 121!.112-128.

Moe. MA.. Jr1989B. Breedingthe ClovI~ Amphiprionocellaris, Part U: A Bit 0' BreedingHistory, Freshwater and Marme Aquarium, 122!:33-38. Moe,M.A., Jr. 1990.The Marine Aquarium Reference: Systems and Invertebrates, Green Turtle Pubhcations, Plantation. FL. Moe,M.A., Jr., 1991.Thoughts on RearmgMarine Organisms. Freshwater and Marine Aquariutn.142!:12-19.

Moe. M.A., Jr.. 1992,The MarineAquarium Handbook. Beginner to Breeder.Green Turtle Pubhcauons. Phmtation, FL. Moe.M.A andF. A. Young,1981. Spewing the Jewelsof the Reef.Freshwater and Marine Aquarium. 4!: 24-84. Meulengracht-MadseLJ., 1971. Breeding Amphiprioa perm'. TropicalFish Hobbyist. 19!:52-57. Pa3co,B.J. and W.J. Richards, 1969. Tbe Rearmg of Cow6shand Related Species From Eggs.Salt Water Aquarist, 5!:67-70. Riley,C.M1994. Captive Spawttmg and Rearing of thePeppermint Shrimp I.vsmara ~emanni!. SeaScope,11:4, Summer. RostiP.. 1967. Breodirq~ theRoyal Qraaama. Salt Water Aquarium. 2!:106-108. Schiller,G., 1990.Spevnmg Behavior of theDecorated Fire6sh lVemateleorris decora!. SeaScope,7:4. FalL ScMler.G., 1991.Spevnmg Behavior of thePuxyie Fire6sh. Marine Fish Moathly. 6 8!:5-6. Schremer,W.. 1972. Breedmg Report-Clowalshes. TheMarine Aquarist. 3!:31-33. SiddaILS,E., 1979A. The Culture of MarineFish Larvae. Part 1. Freshwaterand Marine Aquarium.2 9!:50-53, SiddaR,S.E.. 1979B. The Culture of MarineFish Larvae. Part 1 Continued!.Freshwater andMarine Aquarium.20!:52-53.

PREPARED BY G.~T~ FINAL REPORT - CULTURE OF M 43UNK AQUARIUM FISHES

Sieswerda,P.L.. 1977.Doing What Comes Naturally. The Marine Aquarist, 8!:18-28.

Sowash,D1994. Rearinglysmata. The Breeder'sRegistry, Z!:1-3.

Sprung,J., 1991. Baby Mandarin.Freshwater and Marine Aquarium, 14!:20-24.

Stephens.C., 1988. Those Little Horses of the Sea. Freshwaterand Marine Aquarium, 11!:26-29.

Straughan,R.P.L.. 1959. Salt Water Black Angelfish Spawned.The Aquarium.July:211- 212.

Straughan.R.P., 1968,First Spawningof High Hat. Salt Water Aquarist,4!:73-74.

Strynchuk. J., 1990. An Insight it»o the Mating Habits of Banded Coral Shrimp. Freshwaterand Marine Aquarium. 130!:42-184.

Taylor, E.C., 1973.DascyIhrs arucmm Spawns. Salt Water Aquarium,9!:15-18.

'Buesher, R.E., 1984. Reproductionin Reef Fishes.T.F.H. Pubhcatioas,Neptune City, NJ. 399 pp.

Torben,J., 1992.Comtra'miai Breeding of Pusemonefishes.SeaScope, 9:1%, Wmter.

Tullock, J., 1989, Tank RaisedMarines hmpossible for the Hobbyist?Marine Fish Monthly, 32!;34-35.

Umvin, P.D., 1986. Breeding the YeHow Tailed E~se5sh Glyphiododontops hemicyanea!. SeaScope,3:1, Winter.

Vincent,A., 1995. SeahorseKeeping. The Breeder'sRegistry, 3!:1-5

Waich,J.C., 1993.Breofing Jtwflsh. The Breeder'sRegistry, l!:1-3.

Walch,J.C., 1994. Brce5ag the Royal Gramma,Gnrrnnra lotto. The Breeder'sRegistry, 2!:1-2.

Walker, S., 1977A. Walker SuccessfullySpawn Five Species.Marine Hobbyist News. 5 9!:1-4.

Walker,S,, 1977B. Final ObstacleOvercame. Marine HobbyistNews, 50!:1-9.

Walker.S.D.. 1979. Spawnmgand Rearingthe Neon Goby. Freshwaterand Marine Aquarium,2 8!:40-?.

59 PREPARED BY GADTC FINAL REPORT,- CULTURE OF MAINE AQUAIULUvf FISHES

Walker,S.D., 1991.Reproducmg the Atlantic Spadefishat the Tulsa Zoo. SeaScope,8:1- 2, Winter.

Walker.S.D. and N. Herwig, 1976.Salinity and Spawning,7!:45-50.

Wassink.H. and R. Brons. 1990, A SuccessfulCultivation of the Comet Calloplcsiops alrivelis Steindadmer.1903!. SeaScope,7:1-3, Spring.

Watson, C.A., 1995. Irrvesting in the Future: Captive Breeding of Marine Tropicals. Freshvaterand Marine Aquarium, 18!:160-167.

Weingarten,R.A., 1991. Recertt Developmentsin the Commercial Rearing of Larval MarmeFish. Freshwater and Marme Aquarium, 14 9!:27-208.

WeingarteLR.A., 1992.Tips on RearingNewly SpawnedLarval Marine Fish. Freshwater andMarine Aquarium, 15!:166-212,

Wilkerson, J.D., 1992A. Amphiprion Clanbi: Journey From Egg to Juvenile - Part I, Freshvaterand Marine Aquarium, 150!: 17-21.

Wilkerson,J.D., 1992B.Arnphiprion Clarkii: JourneyFrom Egg to Juvenile- Part II. Freshwaterand Marine Aqtutrium, 152!:154-162.

Wilkerson,J,D., 1993A. ClownmgAround: SimpleClovtmQsh Spawning and Rearing Faci5ties.Freshwater and Mtrine AqaaLritnn,16!:196-206.

Wilkerson,J.D1993B. Spawningand Rotring Anrphiprion Clarkii Clo~~ Aquarium Fish Magazine,5 8!:18-29.

Wilkerson,J,D., 1994.Scarlet Cleaner Shrimp: Care and ReproductiveHabits of Lvsmaut amboincnsis.Freshwater and Maine Aquarium,17 8!:208-215.

Wilkerson.J., 1995.Lys~ ruthbunac.He Breeder'sRegistry, 3!:1-2.

Wilkerson,J., 1996.C-Quest Hatchery: Innovations In t~e OrnamentalMarine Fish Culturmg.Freshwater aud Marme Aquarium, 19!:123-138.

Wirtz,P., 1991.Goby or Blenny?Tropical Fish Hobbyia, XL! ¹428:76-80.

Young,F., 1979,Spawning and Rearing of the BandedCoral Shrimp. Freshwater and Marine Aqua6urn,2!:16-17.

Young,F.A.. 1980.The Spawningand Larval Rearing of the JackknifeFish. Freshwater andh~e Aquarium.5:31-78.

60 PREPARED BY GADTC HNAL REPORT - CiiLTTRtE OF M 4dGNE AQUARKM FTSHES

Young, F.A., 1982.The YellowheadJawfisb. Breeding the Marine Mouthbrooderin Captivity.FreshNmer and Marine Aquariuta. 5!:50-51.

Youag, F.A., 1984. RaisingClo~mfish for the Hobbyist.Freshwater and Marine Aquarium, 7 9!:70-72.

Young, F.A., 1991A. Breediagthe Ormage-FinAaeaeaefish. Marme Fish Monthly, 6!:46.

Young, F,A., 1991B. Rearing the Red Sea Putemonefish,Amphipnon bicinces. Freshwateraad Marme Aquarium, 14!:136-142.

Young, F.A., 1992. Breedmg the Blue~ed Can&mllfish. Freshwater and Marine Aquarium, 15!:36.

Young, F.A., 1993. Artificial Propagation of Spanish Hogfish Bodianus rufus!, Freshwateraad Marine Aquarium. 160!:210-219.

Young, F.A., 1994A.Food Preferencesin Trope& MarineFish Larvae.Aquaculture Magazine JanmIry/Febrtutry!,20 l!:57-59.

Young, F,A., 19948. Live Foods fot Marut Fish Leva». Freshwaterand Marine Aym~ 17!:106-108.

Young,F.A., 1994C.Rearing the GoldenGaby, Elactinus xanthipore. Freshwater and Marine Aquarium, 172!:182-184.

Young,F.A., 1995A. The Porkfish - Its Life History,Ecology, Breeding, and Rearing in Captivity.Freshwater aad Marine Aqmtrima. 18!:189-196.

Young,F.A., 1995B.Reamg Systems for MarineFish Larvae. Freshwater aad Marine Aquarium, 18!:67-70.

Young,F.A., 1995C.Growout Systems for MarineTropical Fish. Freshwater and Marine Aquarium, 18 9!: 150-155.

Youag, F.A., 1996. The State of Tropical Marine AquariumAtmmal Cultivation. Freshwateraad Marme Aqtumaa, 19!:126-129.

61 PREPARED BY GADTC FINAL REPORT - CULTURE OF M

APPENDIX QL Selected Scienti6c Litemtnre on the Culture of Marine Fish and Shrimp

Ako, H., C.S. TamartwP. Bassand C.-S. Lee, 1994, Enhancingthe Resistanceto Physical Stress in Larvae of Mobil cephalic by the Feeding of Enriched Artemia nauplii Aquaculture. 122:81-90.

Alava, V,R. and L.A.O. Gomes, 1989. Breeding Marine Aquarltum Animals: The Anemonefish.Naga, 12!:12-13.

Alayse. J,P,, 1984. Apphcationof techniquesused for temperatemarine fish in breeding Amphiprion ocellaris Cuvier!. Proc.Mar. Aquar. &mnogr. Inst., 10:505-519.

Ballard.J., 1976, BreedingClownfish. South Afi'. Assoc.Mar. Biol. Res., 12:15-17.

Barnabe,G. and A. Guissi, 1994.Adaptations of the FeedingBehavior of Larvae of the Sea Bass, Dicentrarehus labrar L.!, to an Alternating Live-Food/Compound-Food FeedingRegime. Aquaculture and Fisheries Management, 25:537-546.

Benetti, D.D., E.S. Iversenand A.C, Ostrowski, 1994. Growth Ratesof Captive Do1phin. Coryphaeurhippunrs, in Hawaii FisheryBulk%in, 93:152-157.

Blanchard,L., 1992. OrnamentalShrimp Culture - Lysmata grubtumi. Harbor Branch nographicInstitution.

Blancheton,J.-P. and B. Canaguier.Bacteria and ParticulateMaterials in Recirculating Seabass Dice~arehus labour! PrcductionSystem. Aquaculture, 133:21S-224.

Brown, C.L. and B.G. Kim, 1995.Combined Application of Cortisol andTriodothyronine in the Culture of Larval MlrrineFinfish. Aquaculture. 13S:79-86.

Buskey,E.J.. C. Coulterand S. Strom.1993. Locomotory Patterns of Micrmooplanlnon: PotentialEffects on FoodSelom~ of LarvalFhh. Bulletinof MarineScience. 53 1 !:29- 43,

Carh. A., G,L. Mariottini and L. Pane. 1995. Infhence of Nutrition on Fecundity and Survivalin Tigriopusjuly Fischer Copepoda: Harpacricoida!. Aquaculture. 134:113- 119.

Chair.M.. R.S.J.Gapasin. M. Dehasqueand P. Sorgeloos.1994. Vaccination of European SeaBass Fry ThroughBioencapsulation of Artemia Nauplii.Aquaculture International, 2!:254-261.

Chamberlain.G.W., Mget, R.J. and M.G, Haby Eds.!, 1990.Manual on Red Drum Aquaculture.Tech. Rep. Ao. TAMU-SG-90-603,Texas ARM University,236 pp.

PREPARED BY GADTC RNAL REPORT - CULTURE OF 1VL>JUNEAQUA1UU!vf FISHES

Colin,P.L, 1989.P~~ of the Spawningof WestetnAtbaec Butt~4shes Pisces. Chaetodontidae!.Environmental Biology of Fishes.25-3!:131-141. Cotrea.MK.S. Chung and R. Manrique, 1989. [ExpetimetttaiCulture of the SeaHorse, Hippocampuserects.] Cultivo Experimentaldel Caballito de Mar, Hippocampuserectors. BoL Inst. Oceanogr.Venez.. 28-2!:191-196.

Coutteau.P., P. Sorgeloos and P, Lavens. 1994. Progressin the Use of Live Food in Larvicultuteof Marine Fish, Aquacadture,124:283-291.

Craig, S.R., C,R Arnold and G.J. Holt, 1994.Thc KfFectsof EnrichingLive Foods With HighlyUnsaturated Fatty Acids on the Growth and Fatty Acid Compositionof LarvalRed Drum SciaenopsoceOarws. Journal of the World AquacultmeSociety, 25!:424-431.

Crompton,W.D., 1992.Laboratory Culture and Larval Deveiopmetttof the Peppermmt Shrimp,Lysmara wundemanni Gihbes Caridarr Hippoiytidae!.Unpubhshed Masters Thesis,Corpus Christi State University, Corpus Christi. TX.

C~wicz B.S. andC.L. Brown, 1992.Rearing Methods for Two Damse&chSpecies.. DascyilusaSiseQa Giii! andD. amass L.!. Aquaculntre,106:141-149.

DeCastro,A. and D.E. Jory, 1983.PreJiumlry Expmirnerls on the ~ture of the Banded CoralShrimp, Srenopso hispkhs Ohver. J. Aquaticult.Accrue. Sci. 1983,3!:84-89.

Dhert, Ph., Lavens, P., Duray, M. and P. Sorgeloos,1990. IxnprovedLarval Survival at Metamorphosisof Asian Seabass Late caicarifer! Using w3-HUFA-Enriched Live Food. AquaMure, 90:63-74.

Dhert, P., P. Lavens and P. Sorgeloos,1992. State of the Art of Asian SeabassZ.ares caicarifer LarviadlIure.Journal of the World AquacultureSociety, 23!:317-329.

Fertnin. A.C. and M.E.C. Bolivar, 1994. Feeding Live or Frozen Moira macmcopa Strauss! to Asian Sea Bass, hates caicarifer Bloch!, Larvae. The lsraeh Journal of Aquaculture- Bamidgeh,46!:132-139.

Fernandez-Diaz.C.. E. Pascual and M. Yufera. 1994. Feedmg Behavior and Prey Selectionof GiitheadSe&ream, Spans' auruta. LarvaeFed on Inert and Live Food. MarineBiology, 118!:323-328.

Fernaudez-Diaz.C. and M. Yufera, 1995.Capacity of Gilthead Seabream.Spanrs aurara L., Larvaeto Break Down DietaryMicrocapsules. Aquaculture. 134;269-278.

Fletcher. D.J. and M. Wilson. 1996. Vitamm C Requirementsof the AnemoneFish, Amphiprion ciarkii. Page 128 In: Book of Abstracts- World Aquaculture'96. The 1996 AnnualMeeting of the WorldAquaculture Society. Bangkok. Thailand. 498 pp.

63 PREPARED BY GADTC FINAL REPORT,-, CULTUS OF M/UMtE AQUARIL'M FISHES

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PREPARED BY GADTC FlNAL REPORT;,CULTUS OF MAIUNE AQUPdQUMFISHES

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65 PREPARED BY GADTC FATAL REPORT - CULTt3tE OF M/dUNE AQUPdUU154 FISHES

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Sweets, J.W., 1992.Larvicuiture of MediterraneanMarine Fish Species:Current Statusand Future Trends. Journal of the WorMAquacuinm Society, 23!:330-337.

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66 PREPARED BY GAl3TC FINAL REPORT - CULTL'RE OF bbVJNE AQUPdtKM FISHES

Tacbihara.K., R. Ebisu and Y. Tukashima,1993. Spawning, Eggs, Larvaeand Juveniles of the PurphshAmberIack Sen'ola dunrerili. Nippon SuisanGakkaishi. 59 9!:1479-1488.

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Tandier,A., 1993.A4lrine Aquaculturein Israelwith SpecialEmphasis on Larval Rearing. Journalof the Worki AquacultureSociety, 24/2!:241-245.

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67 PREPARED BY GADTC FINAL REPORT - CULTURE OF h4VtINE AQUARIUM FISHES

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68 PREPARED BY GADTC FINAL REPORT- CUL~tE OF hrGVUNE AQUARIUM FISHES

Hioki. S., K. Suzuki and Y, Taaaka, 1990. Developmentof Eggs and Larvae in the Angelfish.Cenrmpyge fenugatus. Japan.J. Ichthyol, 37 l!:34-38.

Leis, J,M., 1989.Larval Biology of Butterfiyfishes Pisces.Chaetodontidae}: What Do We Really Know'?Environ. Bio, Fishes,25-3!:87-100.

Mariiave, J.B,, 1985.Color Polymorphismin SiblingAlnphiprion.. Is the Reef Fish Lottery Rigged?Environ. Biol Fish., 12!;63-68.

Moyer,J.T., 1981,Interspecific Spavmitg of the PygmyAngeifishes Cereepyge shepardi andCentropyge bispinosus at Guam.Micronesica, 17-2!:119-124.

Robertson,D.R., 1983. On the SpmvningBehavior and SpawmagCycles of Eight Surgeoafishes Acandmridae! From the indo-Pacific.Envir. Biol Fishes,9/4!:193-223.

Robertson.D.R.. 1990.DdFerences in the Seasonalitiesof Spewing and Recntitmentof SomeSmail Neotropical Reef Fishes. J. Exp.Mar. Bio. EcoL,144:49-62.

Sakamoto,T. and K. Suzuki, 1978. SpawnmgBehavior and Early life History of the PorcupmePufFer, Diodon holaccmthus, in Aquaria.Jap. 3. Ichthyol,24!-.261-270.

Suzuki.K., Y. Tanakaaat S. Kioh, 1980.Spawning Behaviour, Eggs, and Larvae of the Bufferfiy,Chaeedon nippon, in anAymtum. Jpn.J. Ichthyol,26:334-341.

Tanaka, Y. and K. Suzuki, 1991. Spnaeing, Eggs and bmm of the Hawldish. Cirrhirichthysalbums, in anAquarium. Jap. 3. Ichthyol, 38!:283-288.

Thresher,R.E., 1982.Courtship and Spewing in the EntperorAngelfish Pornacanrhus impemtor, with Conmentson Reproductionby Other PomacanthidFishes. Marine Biology, 70:149-156,

Thresher,R.E., 1984.Reproduction in ReefFish. T.F.H. Publications. Inc. Ltd., Neptune City, N.J,, 399 pp.

Tricas. T.C. and J.T. Kramoto. 1989. Sexual Ddferentiation,Gonad Development.and Spiv~ Seasonahtyof the HawaiianButter&g6sh, Chactodon nndticinctus, Environ. BioL Fish., 25-3!:111-124.

WeHington.M. and B.C. Victor, 1989. PlanktonicLarval Durationof One Hundred Speciesof Pacificand Atlantic Damelfishes Pomacentridae}. Mar. BioL. 101:557-56'T.

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APPENDIX V. List of Marine Aqaariam FishesReareti in Captivity

Species Common Name Abudefduf abdominalis Maomao damsel6sh Abudefduf caery! eus Electric blue damse16sh Abudefdufcyaneus Blue devil damseiBsh Abudefdufsaxatilis Sargeantmajor Acanthostracionquadri cornis Cow6sh Acanthurus leucosternon ~ Powder-bluetang Amblygobiusphalaena Bandedgoby. Brown-barredgoby, Dragongaby Amblygobiusrai nfordi Rainford's goby Amphiprion aiadlopisos Skunk clownish Amphiprion akindivtos Barrier Reef clownBsh Amphiprion allardi AUard's AtMaaoneBsh Amphiprion bicinctus Two-banded clownBsh or Red Sea clowafish Amphiprion chrysopterus Orange-Baclovm6sh or Bine-stripeclownfish Amphiprion clarkii Clarkii clowaBsh Amphiprion ephippium Red saddleback clownish Amphiprionfrenatus Tomato clowaBsh Amphiprion kxrezonatus Wide-band clowaBsh or Lord Howe clown6sh Amphiprion melanopus Ciaaamoa clowafish Amphiprion nigripes Black-Baaed clovm6sh Amphiprion ocellaris Common cloem6sh Amphiprion percula Percuia clo~m6ah Amphiprion perideraion Pink sktrnk clowaBsh Amphipnonpo1ynnus Saddleback clownBsh Amphiprion rubrocinctus Australian clownBsh Amphiprion sandarucinos Orangeskunk clowm6sh Amplg'prionsebae Seba's clannish Amphiprion tricinctus Three-band clowaBsh Anisotrernus vt'rginicus PorkBsh Apogon sp. Gsrdmaifish Apogon compressus Bayed cardiaal6shor Spht-bandedcardiaalBsh Balisandesconspicillum ' Clown triggerBsh Bodianuspulchellus Spot5a HogBsh Bodianusrufus Spanishhog6sh Calloplesiopsaltivelis Comet Marine betta! Chaetodipterusfaber Atlantic spade6sh Chrysipteracyanea Orange-taildamsel Chrysiptera parasema Gold-tail demoiselle Cryptocentrus cinctus Yellow prawn goby Dascyllusalbisela Threespotdamselfish Dascyllus aruanus Whitetailed damselfish Dioa'on holacanthus BalloonBshor porcupiae6sh Equetus acuminatus High-hat

+p PREPARED BY GADTC FlNAL REPORT - CULTURE OF 14LVUNE AQUMUUM HSHES

APFKNDEXV. List of MarbleAquaritrm FbhesReareck in Captivity Coatmued!

Zquetuslanceolatus Jacknife Gsh Equetuspunctatus Spotteddrum Zquetus uInbrosus Cubbyu Glyphidodontopshemicvaneus YeHow-tailed damsel6sh Gobiodon citrinus Citron goby or Clown goby Gobiodon okinm~ Okinawagoby or Lemonor Yeilowclown gaby Gobiosoma evclvnae Shsrknosedgoby Gobiosoma genic Genie'scleanmg goby Gobiosoma macrodon Tiger goby Gobiosomamulttfasciatum ~g le Gobiosomaoceanops Neon goby Gobr'osomapuncticuiatus Redheadgaby Gobiosomaxanthiprora Yellowstripecleaning goby Gramma loreto Royal gramma Gramma meiacara Black cap bas!Set Hippocampuscomes Seahorse Hippocampuserectus Northern seahorse Hippocampusjbscus Seahorse Hippocampushystrix Thorny seahorae Hippocampusltuda Oceamc seahorse HippocampusnovaehoNandiae Anstrlban seahorse Hippocampussp. Lattice seahorse Hippocampussosterue &barf seahorse Hypopiectrusgemma Bhe hamlet Hypopiectrusunicolor Butter hamlet Hypsypopsrubicundus GaribaMi Liopropomarubre Peppermintbasslet I ythrypnusdaNi CataItmagoby Meiacanthus grammistes Stripedblenny Meiacanthusnigrolineutus Black-lined blenny Microspaabodonchrysurus Jewe5sh or Yellow-tailed damse5sh Opistognathusaurifrons Yellowhead]aw6sh Phoiidichdtysleucotaenia Corrvict blenny Phycodurus eques Leafy seadragon PhyNopteryxtaeniolatus Common seadragon Platm orbicuiaris ' Circuhespade6sh or Roundbat6sh Pomacanthus arcuatus Grey angel6sh Pomacanthusimperator ~ Emperorangelfish Pomacanthusparu Frenchangelfish Pomacentrus caendeus Yellow-beHied blue damsel Premnas biaculcatus Maroon clown6sh Pseudochromis dutoiti Neon dottyback Pscudochromisflaviverter Sunrisedottyback

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APPENDIXV. Listof MarineAquarium Hshes Rear' in Captivity Continued!

Pseudochmmisfn'dmani Orch@dattyback Pseudachromispaccagnellae Royaldottyback ar Bicolordattyback PseudochromispoqHryreus Magentadattybacit Pseudochmmis springeri Springer'sdattyback Pterosvnchiropussplendidgs Serranussubligarius Belted saef6sh Siganuscanaliculatus White-spottedspinefoot rabbit6sh! Signigobius biocelkecs ' Signalgoby ar Twa-spot gaby Zanclus cornutus ' Moorish idol ' Reportsof thisspecies being successfuQy cultured need ta be veri6ed.

APPKM!IX VL Listof Marine AquariumShrimp Rear' in Captivity

Scieedic Name Common Name

Lysmataamboinensis Scarletcleaner 0'r Whrte-strTpedcleaner shznrrp Lysmatacalifornica Catalioacleaner shrimp Lysmatadebeiius Bhod or Fireshrimp ar Cardinalcleaner shrimp LysmataratrMerrae Peppermintshrimp Lysmataseticaudata Monaco cleanershrimp Lysmatawurd''emanni Peppermintshrimp ar Caribbeancleaner .Primp Hymenoceru picta Harlequinshrimp ar Ch'an shrimp Rhynehocinetesuritai RosySeeing shrimp Stenopushispid' Bandedcaral shrimp

72 PREPARED BY GADTC arieie.KA Glar OeostlpRY PELtLIBRARY BgtLDfffg " ~I~~R<":AHSL,.6AVCaMPUS "PP