Inoculating SHB larvae with entomopathogenic fungi

Fungal infected cadavers (Ellis et al., 2004e) or conidial suspensions (Muerrle et al., 2006) can be used to infect SHBs. Ellis et al. (2004e) achieved about 30% mortality among pupating beetles exposed as wandering larvae to beetle cadavers from which Aspergillus flavus and A. niger were isolated (see BEEBOOK paper by Jensen et al., 2013):

  1. Collect healthy-looking wandering SHB larvae produced using autoclaved soil and the rearing method (see section 3.1.2.).
  2. Place desired amount of wandering larvae in a small plastic container [11 x 11 x 9 cm] that has SHB larvae/pupae that show signs of being infected with a fungal pathogen (Ellis et al., 2004e). It is important to confirm that the cadavers are carrying the pathogen of interest using adequate microbiological techniques (Mürrle et al., 2006 and the respective BEEBOOK paper (Jensen et al., 2013)).
  3. Allow the healthy larvae to wander in the container among the cadavers for 24 hours.
  4. After 24 hours exposure to infected cadavers, place the wandering larvae in soil pupation chambers and allow to pupate (see section 3.1.2.).