3.1.7.2.4. In vitro soil infestation of pupae with nematode-infected cadavers

Follow the method outlined in section 3.1.7.2.3. with one modification at step 5.

5. Rather than applying nematodes via aqueous suspension, add a SHB cadaver infected with the target nematode to the cup.

Bury the cadaver about 0.5 cm below the soil surface. The cadaver can be produced according to the method outlined in section 3.1.7.1.

Notes:

  • The methods outlined can be scaled up and modified for field use.
  • Ellis et al. (2010) did this by burying the soil cups outside. The cups had screened lids and bottoms to allow rain drainage through the cups. Otherwise, nematodes and SHB larvae were added to the cups as in the in vitro assays. The methods also can be used to determine the generational persistence of the nematodes. This is accomplished by adding 5 additional SHB larvae to the soil cups weekly. If the nematodes are reproducing and moving into the newly-burrowing larvae, adult SHB emergence rates in the soil cups will be perpetually low.