5.4. Comb treatment

The compound(s) of interest can be directly incorporated into melted wax prior to mill rolling of foundation sheets or applied to previously milled foundation (Burges and Bailey, 1968; Burges, 1976; Vandenberg and Shimanuki, 1990; Hood et al., 2003, Ellis and Hayes, 2009).

  1. Application to wax comb foundation: Based on the available form of the compound of interest, it may be sprayed, dipped, aerosolized, or dusted onto previously milled foundation sheets per the needs of the experiment. In the past, fogging (or aerosols) has been showed to be a less effective method for effective application - Vandenbergi and Shimanuki (1990).
  2. Once dried, use treated foundation in experiments as is or sandwiched between untreated sheets of foundation and remilled to prevent direct exposure of honey bees in the colony to the test compound in the treated foundation.
  3. Insert frames containing treatment and control foundation into healthy colonies for comb construction. The colonies may need to be fed a sucrose solution to encourage bees to construct comb on the foundation.
  4. Once drawn, remove the experimental frames from the nest and any honey extracted from the comb.
  5. Place newly hatched wax moth larvae (reared per Section 3) singly on a small piece of treated or control comb (comb produced on untreated foundation) in a dish container,
  6. Incubate at 34°C,
  7. Monitor for physiological changes.

Alternatively, hive boxes containing frames of treated comb, but no bees, can be inoculated with wax moths and the level of damage assessed (per Section 4).

Considerations: Compound concentrations should be determined for drawn comb after removal from the colony as honey bees will distribute wax from foundation throughout the comb  (Burges and Bailey, 1968). Test compound impacts on honey bee colony fitness and/or behaviour should be accessed. Recommended methods for measuring colony strength parameters can be found in the BEEBOOK paper on measuring colony strength parameters (Delaplane et al., 2013). If incorporating the compound directly into melted wax prior to milling, one must know heat impacts on the compound. The average temperature used to melt wax for milling machines has been reported to range from 77° to 99°C (Burges and Bailey, 1968).