Disinfestant solution: 0.5% chlorhexidine diacetate (hibitane)
- dissolve 0.5 g into 100 ml sterile distilled water
- dissolve 2.5 g into 500 ml sterile distilled water
- Concentrate eggs in 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 (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 (as in Step 1), pour off the water, add fresh sterile distilled water and incubate one final time for 5 minutes.
- Centrifuge to concentrate the eggs, pour off the water and decant the surface disinfested eggs into a suitable container.