3.1.7.2.3. In vitro soil infestation of pupae with an aqueous suspension of nematodes Autoclave soil.

  1. Moisten soil to 10% water by weight.
  2. Place autoclaved soil in small plastic cups.
    Ellis et al. (2010) used 118 ml cups. A similar size is appropriate.
  3. Place 5 wandering SHB larvae in the soil cups and allow them to bury themselves and begin the process of pupating.
  4. Two days after the addition of larvae, pipette the desired amount of IJs suspended in an aqueous solution onto the soil in the cup.
  5. Add 5 ml of water to the soil cups every 3-4 days as needed to maintain adequate soil moisture.
  6. Place the cup lids on the cups to enclose the SHBs.
  7. One can expect the SHBs in the control cups (no nematodes) to begin to emerge as early as day 17 post addition to the soil.