Passing

  1. Prepare 45-50 g of Plodia diet in a modified container (copper mesh soldered to the lid for ventilation). Pack it tightly down to the bottom of the container with a gloved fist.

  2. Measure 10-12 mg of Plodia eggs on a small weighing boat by tapping the metal cup gently.

    <aside> 💡 Plodia eggs disperse easily by sudden air change in airflow (through breathing or moving them too fast) and electrostatic effects (dry plastic, nitrile gloves). They are still used during handling but make sure to kill any residue especially on the gloves, the bench, and the weighing scale if the eggs do disperse.

    </aside>

  3. Place the weighing boat on top of the packed food.

  4. Cover the copper mesh of the stock container with filter paper and tape around it for label.

  5. Information on the label: Pi_(strain) [date passed] [10-12 mg eggs]/[45-50 g diet]

  6. Leave at 28'C until they hatch and become larvae.

  7. Check every 3-4 days for any signs of infection (yeast), food depletion (overly wandering larvae), or overproduction of silk.

    <aside> 💡 Optional: Pupal isolation can be done around 15 days after egg laying. Place some cut cardboard for the larvae to pupate in ("hotels"). Separate them as individual manually (e.g. for sexing) or as hotels into another container (e.g. to rescue from mold infection).

    </aside>

  8. At around 25-28 days, adults should have emerged. Leave freshly emerged adults 1-2 days to mate then start the synchronized oviposition procedure.

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Synchronized Oviposition

<aside> 💡 Make sure the inside of the mason jar is completely dry. Eggs may adhere to damp spots inside the jar.

</aside>

  1. Prepare a clean mason jar with mesh lid as oviposition jar, stock containers with adults, ethanol, and a tray as a work surface.

  2. Anesthesize Plodia adults using CO2 gas through the modified container.

  3. Tap down the moths to one side lightly as not to disturb the food bedding.

  4. Open the container and transfer moths (min. 30 moths) into the mason jar. Work quickly as they will wake up in about 10-20 seconds.

  5. Leave the moths in the oviposition jar in the dark at 28'C for 4 hours (to overnight) to mate and lay eggs. Save the adults in the stock container as backup.

    <aside> 💡 Adults should not be left inside more than three days since the mesh is larger than the copper mesh used in the stock container. Earlier eggs might hatch as larvae and escape the steel mesh.

    </aside>

  6. Once enough eggs are laid on the bottom of the mason jar, collect them by tapping the mason jar upside down into a metal cup (to prevent static) through the mesh lid.