1. Zebrafish, Danio rerio, Husbandry Round Table Discussion Zebrafish Development and Genetics MBL 2012

2. Zebrafish International Resource Center Central Repository Central Repository Distribution of Lines & Reagents Distribution of Lines & Reagents Health & Husbandry Services Health & Husbandry Services University of Oregon

3. Environmental and Water Quality Parameters Lighting: 14 hours/day Lighting: 14 hours/day Temperature: 28.5˚C (22-30˚C tolerated) Temperature: 28.5˚C (22-30˚C tolerated) pH: 7.2 to 7.6 pH: 7.2 to 7.6 Conductivity: 500 -1500 µS Conductivity: 500 -1500 µS Ammonia: 0 ppm Ammonia: 0 ppm Nitrite: 0 ppm Nitrite: 0 ppm Nitrate: < 10 ppm Nitrate: < 10 ppm Exchange rate: 10% per day (RO water) Exchange rate: 10% per day (RO water)

4. Feeding Larval Diets Larval Diets Dry - micro-capsule Dry - micro-capsule Live - Paramecia, Artemia naupli, rotifers Live - Paramecia, Artemia naupli, rotifers Adult Diets Adult Diets Dry - Flake, pellet, crumble, freeze dried Dry - Flake, pellet, crumble, freeze dried Live - Artemia (brine shrimp) Live - Artemia (brine shrimp) Varied diet recommended Varied diet recommended 2-4 feedings/day 2-4 feedings/day Do Not Overfeed Do Not Overfeed

5. Larval Culture Nursery care 3+ weeks Nursery care 3+ weeks Feeding based on larval size Feeding based on larval size Water changes Water changes Hand siphoning Hand siphoning Continuous flow Continuous flow Lab specific protocols Lab 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 fed Day 5: transferred to tanks and fed (paramecia, rotifers, powdered food, artemia, or other) (paramecia, rotifers, powdered food, artemia, or other) Day 5-9: <100 to 250 micron food Day 5-9: <100 to 250 micron food Day 10-21: 100 to 500 micron food Day 10-21: 100 to 500 micron food Day 21-25: transferred to aquariums, 150 to 500 micron food Day 21-25: transferred to aquariums, 150 to 500 micron food

6. Breeding Natural Crosses Natural Crosses Pairs or small groups Pairs or small groups Embryo collection Embryo collection Special breeding tanks Special breeding tanks Marbles Marbles In Vitro Fertilization In Vitro Fertilization Synchronized development of embryos Synchronized development of embryos Do Not Over Breed Do Not Over Breed Allow 1-2 weeks between breeding events Allow 1-2 weeks between breeding events

7. Biosecurity Quarantine All fish from outside sources All fish from outside sources Isolated from central facility Isolated from central facility Tanks, racks, water system Tanks, racks, water system Equipment, nets, glassware, etc. Equipment, nets, glassware, etc. Observation of adult fish Observation of adult fish Observe adults 2-3 weeks prior to breeding Observe adults 2-3 weeks prior to breeding Euthanatize or treat sick fish Euthanatize or treat sick fish Surface sanitize embryos Surface sanitize embryos Highly recommended Highly recommended

8. Internal Biosecurity General Practices General Practices Remove sick and dead fish ASAP Remove sick and dead fish ASAP Minimize transfer of fish and water Minimize transfer of fish and water between tanks between tanks Avoid disease spread from breeding/ Avoid disease spread from breeding/ out-crossing to wild-types out-crossing to wild-types Use nets in one tank only Use nets in one tank only Sanitize equipment between use Sanitize equipment between use Sentinel Fish Sentinel Fish Exposure to return (dirty) water Exposure to return (dirty) water Observed daily Observed daily Histopathology every 3-6 mo. Histopathology every 3-6 mo.

9. Contact Information Zebrafish International Resource Center University of Oregon Web Address: www.zebrafish.org www.zebrafish.org April Freeman, Facility Manager: april@zebrafish.org General Email: zirc@zebrafish.org zirc@zebrafish.org ZIRC is supported by grant P40 RR12546 from the NIH-NCRR. Tufts University Boston Caitlin Stewart-Swift, Zebrafish Facility Supervisor: Caitlin.Stewart_swift@tufts.edu