5.3.1. Larvae

  1. Mix purified virus of the appropriate minimum concentration (Table 3) with medium for in-vitro larval rearing and allocate this to the wells of a 48-well tissue culture plate (see the BEEBOOK paper on in-vitro larval rearing (Crailsheim et al., 2013).
  2. Transfer two-day old larvae to the wells, making sure there is enough virus so that each larva gets the minimum infectious dose.
  3. Follow the procedures for in-vitro larval rearing (see the BEEBOOK paper on in-vitro larval rearing (Crailsheim et al., 2013), transferring the larvae periodically to fresh food, either including or excluding further virus extract.
  4. Include a series of control inoculations, using larval food medium without virus.