Transcript Zebrafish, Danio rerio, Husbandry, Egg to Egg, the Do`s and Don`ts
Zebrafish, Danio rerio, Husbandry
Round Table Discussion Zebrafish Development and Genetics MBL 2012
Zebrafish International Resource Center
• Central Repository • Distribution of Lines & Reagents • Health & Husbandry Services
University of Oregon
Environmental and Water Quality Parameters
• • • • • • • •
Lighting: 14 hours/day Temperature: 28.5˚C (22-30˚C tolerated) pH: 7.2 to 7.6
Conductivity: 500 1500 µS Ammonia: 0 ppm Nitrite: 0 ppm Nitrate: < 10 ppm Exchange rate: 10% per day (RO water)
Feeding
•
Larval Diets
• Dry - micro-capsule • Live - Paramecia,
Artemia
naupli, rotifers •
Adult Diets
• Dry - Flake, pellet, crumble, freeze dried • Live -
Artemia
(brine shrimp) • Varied diet recommended •
2-4 feedings/day
• Do Not Overfeed
Larval Culture
• • • •
Nursery care 3+ weeks Feeding based on larval size Water changes
• Hand siphoning • Continuous flow
Lab specific protocols
• Day 0 to 4: 25-50 embryos per dish • Day 5: transferred to tanks and fed (paramecia, rotifers, powdered food, artemia, or other) • Day 5-9: <100 to 250 micron food • Day 10-21: 100 to 500 micron food • Day 21-25: transferred to aquariums, 150 to 500 micron food
Breeding
• • •
Natural Crosses
• • Pairs or small groups Embryo collection • • Special breeding tanks Marbles
In Vitro Fertilization
• Synchronized development of embryos
Do Not Over Breed
• Allow 1-2 weeks between breeding events
Biosecurity Quarantine
• • • • •
All fish from outside sources Isolated from central facility
• • Tanks, racks, water system Equipment, nets, glassware, etc.
Observation of adult fish
• Observe adults 2-3 weeks prior to breeding
Euthanatize or treat sick fish Surface sanitize embryos
• Highly recommended
Internal Biosecurity
• •
General Practices
• Remove sick and dead fish ASAP • Minimize transfer of fish and water between tanks • Avoid disease spread from breeding/ out-crossing to wild-types • • Use nets in one tank only Sanitize equipment between use
Sentinel Fish
• Exposure to return (dirty) water • Observed daily • Histopathology every 3-6 mo.
Contact Information
Zebrafish International Resource Center University of Oregon Web Address: www.zebrafish.org
April Freeman, Facility Manager: [email protected]
General Email: [email protected]
ZIRC is supported by grant P40 RR12546 from the NIH-NCRR.
Tufts University Boston Caitlin Stewart-Swift, Zebrafish Facility Supervisor: [email protected]