Surface disinfestation of SCN eggs

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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.