To prepare the disinfestant solution of 0.5% chlorhexidine diacetate (Hibitane), dissolve 0.5 g into 100 ml sterile distilled water, or dissolve 2.5 g into 500 ml sterile distilled water
To disinfest the eggs:
- Concentrate the eggs in a 50-ml centrifuge tube by centrifuging at 2,000 RPM for 3 minutes.
- Under the laminar flow hood, pour off the water (taking care not to disturb the pellet of eggs), and add the Hibitane solution.
- Cover the tube opening with a square of Parafilm, shake to resuspend the egg pellet, and incubate at room temperature for 15 minutes.
- Centrifuge the egg suspension again (as in Step 1) to concentrate the eggs and pour off the Hibitane solution while working in the hood.
- Add sterile distilled water to the tube, cover the tube, and shake to resuspend the eggs, then incubate for 10 minutes.
- Centrifuge the egg suspension, pour off the water, add fresh sterile distilled water and incubate one final time for 5 minutes.
- Centrifuge a fourth time to concentrate the eggs, pour off the water and decant the surface-disinfested eggs into a suitable container.