126.96.36.199.3. In vitro soil infestation of pupae with an aqueous suspension of nematodes Autoclave soil.
- Moisten soil to 10% water by weight.
- Place autoclaved soil in small plastic cups.
Ellis et al. (2010) used 118 ml cups. A similar size is appropriate.
- Place 5 wandering SHB larvae in the soil cups and allow them to bury themselves and begin the process of pupating.
- Two days after the addition of larvae, pipette the desired amount of IJs suspended in an aqueous solution onto the soil in the cup.
- Add 5 ml of water to the soil cups every 3-4 days as needed to maintain adequate soil moisture.
- Place the cup lids on the cups to enclose the SHBs.
- 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.