Hatching protocol from my lab
Hey macd,
Below is a "hatching" protocol I followed for my fish lab. I've probably "hatched" out 100 or more batches and each batch always had a successfully high yield. Of course you'll have to find some way to get a hold of some NaOH. If not, you may still be able to do the step with the bleach alone but I'm not sure. This step helps remove the cyst, which can prevent many brine shrimp from "hatching." Low yields can also be the result of decreased flow. It is always a bad idea to set up the hatcher in such a way that the eggs lay on the bottom of the hatcher. Ideally you want a high air flow which creates a current that constantly pulls the eggs up from the bottom of the hatcher...like an upside down 2 liter coca-cola plastic bottle. All of the items were purchased from Aquatic Habitats, except the NaoH (Fisher Scientific), which may be hard for an individual to purchase. This protocol will be hard to follow if you don't have some type of net/mesh/sieve that will allow you to rinse the cysts. A brine shrimp net will not work. A lot of cysts will be lost. I also agree with others that some cysts are higher quality than others which means they haven't been sitting around as long. Try to find some cysts online if the protocol here doesn't work. Sorry to ramble on so much. Cheers!
1) When handling the hatcher be very careful to NEVER put any unnecessary pressure on the stop valve protruding from the bottom of the hatcher.
2) Rinse the hatcher out thoroughly with the stop valve open, sponging off the sides to remove slime from previous hatching. NEVER use soap of any kind to clean the hatcher. If the hatcher is not thoroughly cleaned, hatching rates will decline.
3) Fill the hatcher with water from system 1, 2, or 3 approximately four inches from the top. Then place the hatcher inside the holder and aerate.
4) Place 6 teaspoons of sea salt (approximately 35 grams) in a 600ml beaker and fill with water from system 1, 2, or 3. Take solution with a stir bar inside and place on the stirrer. Turn the right knob to somewhere between 8 and 9 and let the solution mix. Make sure NOT to turn the left knob.
5) While the salt water is mixing, put 100ml of R.O. water in a 250 ml beaker. Then add 50 ml of bleach and 1.5 grams of NaOH.
6) Once the salt has COMPLETELY dissolved in the water, pour the salt water into the hatcher and continue to aerate the hatcher. Remove the stir bar from the 600 ml beaker and put it in the 250 ml beaker. Place the 250 ml beaker on the stirrer to mix the water, bleach and NaOH.
7) When the NaOH is completely dissolved and you are certain you’ve added the correct amount of ingredients, measure out and add 7 grams of eggs to the solution. Stir this mixture for 10 minutes on the stirrer (setting the right knob between 8 and 9) but don’t just sit around and wait for ten minutes . . . immediately begin the next step so you are ready to drain and rinse the eggs after ten minutes.
8) While waiting, fill a plastic 1000 ml nalgene flask with water from system 1, 2, or 3 and locate the rotifer sieve labeled “bleach.” After ten minutes, pour the 250 ml beaker into the rotifer sieve labeled “bleach.” Be careful during this step NOT to splash this solution which contains bleach. It will damage your clothes and could damage your vision if you aren’t careful. Use the water in the flask to help rinse out the remainder of the eggs in the 250 ml beaker and then pour the rest of the water slowly over the eggs in the sieve while swirling the mixture the whole time.
9) Once the water is completely used up, take the sieve and dip it sideways into the hatcher, allowing water to come into it. Then pour the water back into the hatcher. Continue doing this until the majority of the eggs are no longer in the rotifer sieve and are floating in the hatcher.
10) Be certain that the pump is aerating the hatcher correctly and make any necessary adjustments to the hard tube in the hatcher so that it is near, but not in the center of the hatcher. It is very important that the eggs do not come to a rest on the bottom of the hatcher. Check the hatcher 10 or 20 minutes later to be certain that the eggs are circulating around the hatcher and not just resting on the bottom. If the eggs come to rest on the bottom of the hatcher, hatching rates will decline drastically.