This protocol was adapted from the Bruce lab at the University of Toronto (Thank you, Ashley Bruce!)
Raising new generations
Raising Babies
The key to healthy adults starts at fertilization--keeping the embryos as clean as possible and do not crowd them
Day 0
1st 2-3 weeks
Bleaching Embryos
Raising new generations
- New generations should be raised approximately every 8 months, so that the next generation is ready to breed when the old one has reached 1 year of age.
- For problem fish, new generations should be raised every 6 months
- Once the new generation is breeding regularly, the old generation should be euthanized.
Raising Babies
The key to healthy adults starts at fertilization--keeping the embryos as clean as possible and do not crowd them
- Raising healthy adults requires time, effort, patience and care—there are no shortcuts
- Your babies are your responsibility
- All fish facility users are expected to adhere to the following procedures
Day 0
- The day the embryos are laid.
- Collect, keep them in a glass dish or bowl in fresh embryo medium or facility water in the incubator.
- During day zero it is very important to remove ALL abnormal, unfertilized embryos and other debris (poop, excess food etc) as soon as possible.
- If you do not remove sick/dead embryos and other junk, the water will become polluted and the rest of the embryos will die.
- Bleach on the morning of day 1 and dechorionate later that day.
- Remove ALL chorions from the water. The chorions will rot if not removed which will foul the water and kill the larvae.
- No more than 50 embryos per petri dish. They will not develop properly if over-crowded.
- Keep in incubator.
- Check larvae daily.
- Remove abnormal embryos and other crud daily.
- Remove some embryo medium and replace with fresh embryo medium daily (every other day from day 4 onwards).
- On day 7 larvae can be transferred to the facility and get their first meal (food 0)
- No more than 30 larvae per baby tank.
- Putting more than 30 larvae in a baby tank is a really bad idea—they will NOT do well and you will have wasted your time.
- Put tape for WATER OFF on the tank.
1st 2-3 weeks
- Check larvae regularly and remove any dead ones and any excess food.
- Baby tanks should be changed if a smell starts to develop or if there is a lot of debris and/or corpses on the bottom of the tank.
- Over-feeding will not help your larvae grow, the food will rot, foul the water and kill the larvae.
- Check the size of fry—when they look large enough give them a drop of brine shrimp to see if they can eat it.
- You must regularly monitor the fish and increase the food size as appropriate! Once they are 2 couple of weeks old, switch the tape to DRIP or SLOW DRIP.
- Once ALL the larvae are eating brine shrimp, move off the baby shelf into a larger tank with fine mesh in the back and to food #1 + BS (brine shrimp). If using 2L tanks place no more than 20 larvae per tank. Once the larvae are a month old they should be split down to not more than 10-12 larvae per tank.
- Fine meshes should be used for fish eating Food 1. Larger mesh should be used when fish are eating Food 2. When fish are eating adult the mesh should be removed and the babies moved out of the juvenile area.
Bleaching Embryos
- Note that you can use facility water for bleaching—move them to embryo medium after dechorionation.
- This is ideally done the on the morning of day 1. It can also be done on day zero, after gastrulation is complete (10 hours post-fertilization).
- Fill 4 finger bowls halfway with facility water.
- Add 100-200 µL of 6% bleach into the first finger bowl.
- Place the embryos in a silver mesh holder and incubate in the bleach water for 3 min.
- Remove the silver mesh holder (with the embryos) from the bleach water and incubate in facility water for 3 min. Repeat this 2 more times in fresh facility water.
- Transfer the embryos to a new petri dish containing facility water.
- Embryos should be dechorionated that same day.