SOP Amendments

Artemia Setup Protocol pH Adjusted

The Brine

Fill the artemia hatchery with 7.0 L RO water.

Add 700 ml of salt solution, 5 ml of ammo-lock, and 5 ml of Ca(OH)2.

Set the air supply so that it is strong enough to keep all of the cysts in suspension but not so vigorous that it splashes cysts up on to the side of the hatchery.

Decapsulation

Weight out 18 grams of cysts from the freezer in to a 1.0 L beaker. Add 250 ml of sodium hypochlorite (bleach) to the beaker and go to the hot plate in H225.

Behind the hotplate there are some stir bars and tweezers. Place a stir bar in to the beaker and cover the beaker with aluminum foil. Place it on to the hot plate and turn the hot plate on. Make sure the temperature and stir dial are initially set at 0. Slowly turn the stir dial up to 2. LEAVE THE HEAT OFF!

Start the timer above the hot plate. Mix for 4 minutes no more no less. After 4 minutes turn off the hot plate. Remove the stir bar with a pair of tweezers and foil. Transfer the cysts to the artemia net and rinse vigorously with system water for 1 minute.

Place cysts in to hatchery.

Rotifer Feed (Marine)

Feed increased to 15,000/1.0L per feeding for a total of 30,000/day.

Keep carboy cultures green with low to moderate aeration.

Refill carboy with 5ppt water before it drops below 5.0L mark.