Taking care of eggs

Preparing eggs for hibernation:

• 1. Prepare a 2% bleach solution to clean/sterilize the eggs. To make 1 L of the 2% bleach solution combine 20 ml of standard bleach with 980 ml of dH20. You will use about 50-100 ml per set of eggs.

  2. Clean the eggs from each female (which should be in their own petri dish or tube) separately. Transfer the eggs to a vacuum filter (use a damp paint brush for this). Pour 25-50 ml of the 2% bleach solution in with the eggs. Turn on the vacuum to suck the liquid through. Repeat this process. Now, do this two more times but with dH20 rather than the 2% bleach solution.

  3. Transfer the eggs to a small, labelled petri dish. Place one half of the petri dish in a larger petri dish. Label the large petri dish too. Add water to the large petri dish. You want to keep water in the large petri dish but don't want the small petri dish to float around too much. Let the eggs dry for a few minutes than close and seal (e.g., using a rubber band) the large petri dish.

  4. Collect all of the washed eggs in a series of containers so that subsets can be pulled out for maintenance.

  5. Store the eggs in growth chamber at 4C with no light (constant dark), and with humidity plug in.

Weekly maintenance:

• 1. Remove one container of petri dishes/eggs from the growth chamber at a time. Do not leave the door to the growth chamber open any longer than necessary and try to minimize the time eggs are out of the growth chamber.

  2. Carefully open each petri dish and inspect the eggs and conditions. Leave each petri dish open for ~1 minute to air out.

  3. Add more water to outer petri dish if needed (i.e., if it is dry or will likely dry out before the next scheduled maintenance).

  4. Remove any eggs that show signs of fungal infection or rot using a pain brush. Dip the paint brush in a 2% bleach solution, use it to remove the egg, and then wipe the are immediately around the removed egg with 2% bleach followed by dH20 (this should prevent/minimize the spread of rot/fungal growth).

  5. Remove, wash, and disinfect (2% bleach) any petri dishes that have no viable eggs remaining.

  6. Keep track of maintenance on a calendar.