Zebrafish, Zebrafish, Danio rerioDanio rerio, , HusbandryHusbandry

Round Table DiscussionRound Table Discussion

Zebrafish Development and GeneticsZebrafish Development and Genetics

MBL 2012MBL 2012

Zebrafish International Resource CenterZebrafish International Resource Center

• Central RepositoryCentral Repository

• Distribution of Lines & Distribution of Lines &

ReagentsReagents

• Health & Husbandry Health & Husbandry

ServicesServices

University of OregonUniversity of Oregon

Environmental and Water Environmental and Water Quality ParametersQuality Parameters

• Lighting: 14 hours/dayLighting: 14 hours/day

• Temperature: 28.5˚C (22-30˚C tolerated)Temperature: 28.5˚C (22-30˚C tolerated)

• pH: 7.2 to 7.6pH: 7.2 to 7.6

• Conductivity: 500 -1500 µSConductivity: 500 -1500 µS

• Ammonia: 0 ppmAmmonia: 0 ppm

• Nitrite: 0 ppmNitrite: 0 ppm

• Nitrate: < 10 ppmNitrate: < 10 ppm

• Exchange rate: 10% per day (RO water)Exchange rate: 10% per day (RO water)

FeedingFeeding

• Larval DietsLarval Diets• Dry - micro-capsule Dry - micro-capsule • Live - Paramecia, Live - Paramecia, ArtemiaArtemia

naupli, rotifersnaupli, rotifers• Adult DietsAdult Diets

• Dry - Flake, pellet, crumble, Dry - Flake, pellet, crumble, freeze driedfreeze dried

• Live - Live - ArtemiaArtemia (brine shrimp) (brine shrimp)• Varied diet recommendedVaried diet recommended

• 2-4 feedings/day2-4 feedings/day• Do Not OverfeedDo Not Overfeed

Larval CultureLarval Culture

• Nursery care 3+ weeksNursery care 3+ weeks• Feeding based on larval sizeFeeding based on larval size• Water changesWater changes

• Hand siphoningHand siphoning• Continuous flowContinuous flow

• Lab specific protocolsLab specific protocols• Day 0 to 4: 25-50 embryos per dish Day 0 to 4: 25-50 embryos per dish • Day 5: transferred to tanks and fedDay 5: transferred to tanks and fed

(paramecia, rotifers, powdered food, (paramecia, rotifers, powdered food, artemia, or other)artemia, or other)

• Day 5-9: <100 to 250 micron foodDay 5-9: <100 to 250 micron food• Day 10-21: 100 to 500 micron foodDay 10-21: 100 to 500 micron food• Day 21-25: transferred to Day 21-25: transferred to aquariums, 150 to 500 micron foodaquariums, 150 to 500 micron food

BreedingBreeding

• Natural CrossesNatural Crosses• Pairs or small groups Pairs or small groups • Embryo collectionEmbryo collection

• Special breeding tanksSpecial breeding tanks• MarblesMarbles

• In VitroIn Vitro Fertilization Fertilization• Synchronized development Synchronized development

of embryosof embryos

• Do Not Over BreedDo Not Over Breed• Allow 1-2 weeks between Allow 1-2 weeks between

breeding eventsbreeding events

BiosecurityBiosecurityQuarantineQuarantine

• All fish from outside sourcesAll fish from outside sources

• Isolated from central facilityIsolated from central facility

• Tanks, racks, water systemTanks, racks, water system

• Equipment, nets, glassware, etc.Equipment, nets, glassware, etc.

• Observation of adult fishObservation of adult fish

• Observe adults 2-3 weeks prior to Observe adults 2-3 weeks prior to

breedingbreeding

• Euthanatize or treat sick fishEuthanatize or treat sick fish

• Surface sanitize embryosSurface sanitize embryos

• Highly recommendedHighly recommended

Internal BiosecurityInternal Biosecurity

• General PracticesGeneral Practices• Remove sick and dead fish ASAPRemove sick and dead fish ASAP• Minimize transfer of fish and water Minimize transfer of fish and water between tanksbetween tanks• Avoid disease spread from breeding/ Avoid disease spread from breeding/ out-crossing to wild-typesout-crossing to wild-types• Use nets in one tank onlyUse nets in one tank only• Sanitize equipment between useSanitize equipment between use

• Sentinel FishSentinel Fish• Exposure to return (dirty) water Exposure to return (dirty) water • Observed dailyObserved daily• Histopathology every 3-6 mo.Histopathology every 3-6 mo.

Contact InformationContact Information

Zebrafish International Resource CenterZebrafish International Resource CenterUniversity of OregonUniversity of Oregon

Web Address: Web Address: www.zebrafish.orgApril Freeman, Facility Manager: April Freeman, Facility Manager: april@zebrafish.orgapril@zebrafish.org

General Email: General Email: zirc@zebrafish.org

ZIRC is supported by grant P40 RR12546 from the NIH-NCRR.ZIRC is supported by grant P40 RR12546 from the NIH-NCRR.

Tufts University Tufts University BostonBoston

Caitlin Stewart-Swift, Zebrafish Facility Supervisor: Caitlin Stewart-Swift, Zebrafish Facility Supervisor: Caitlin.Stewart_swift@tufts.eduCaitlin.Stewart_swift@tufts.edu