In-Vitro Hatching Studies

Protocol

Procedure:

  • Construct microsieves from 18-mm and 20-mm test tube caps and 25 micrometer pore nylon mesh.
  • UV sterilize all microsieves and hatch trays.
  • Arrange hatching trays in replicated boxes using pre-generated random order.
  • Fill each hatching tray with 12 ml of the appropriate test or control solution.
  • Pipette between 5,000 and 10,000 (depending on experimental details) surface disinfested SCN eggs onto each microsieve.
  • Incubate the experiment at room temperature in darkness.
  • At every-other day intervals, transfer the microsieves to duplicate hatching trays containing fresh solution, and count the hatched second-stage juveniles remaining in the original solution.