We suggest 4 AMPs for the treatment of Melissococcus plutonius and Paenibacillus larvae . Defensin 1, Defensin 2, Apidaecin and Abaecin are suggested AMPs. These AMPs possess different mechanisms of action. The defensins depolarize the membrane, open channels in it, and allow the efflux of potassium ions, which would either be compensated with contaminating cations or efflux of anions, destabilizing biochemical processes. There also is evidence that defensins stop respiratory activity and reduce the level of ATPs present. Abaecin and apidaecin, on the other hand, stop protein synthesis through interference of the 70S ribosome and inhibits DnaK activity. Apidaecin binds to LPS and disrupts the ABC transport system, as well.

The project’s objective is to express these AMPs in a different bacterial culture each, with an inducible promoter. T7 RNA polymerase expression is regulated by the Lac operon, which is induced through the presence of IPTG. Therefore, the usage of T7 promoters will allow us to induce AMP production. Proteins will be extracted through sonication of bacterial cultures and then isolated through a His-tag purification process.

Once all four AMPs are purified, all possible combinations will be tested against P. larvae and M. plutonius . A 2⁴ factorial design will be used to evaluate individual effects, quantify interactions, and identify the optimal AMP combination. PLGA-microencapsulated AMPs would be released in the bees’ diet so nurses can bring the AMPs to the infected larvae. This final product would be available for beekeepers to apply in their beehives through internal or external feeding mechanisms and thus inhibit the proliferation of the pathogenic bacteria.