Detection and isolation of bacteriophage from Paenibacillus larvae

Abstract

American Foulbrood is currently one of the most widespread and devastating brood diseases in honeybees. It has caused huge financial losses in apiculture businesses, with additional effects on crop pollination and productivity. Traditionally, treatments for American Foulbrood disease have employed the antibiotic approach. However, the increasing issues of resistant bacterial strains and antibiotic residues in honey have promoted research towards alternative treatment options. This study focused on detecting and isolating bacteriophage of Paenibacillus larvae. The primary method pursued was to identify phage within current stocks of bacteria through PCR detection methods. Several DNA extraction protocols were assessed to provide material for PCR detection. Pairs of primers to select for four known and previously characterized P. larvae bacteriophages were used. Best results were obtained using philBB_P123 primers, after an extensive DNA extraction process involving filtration of culture solutions, and heat and cold treatments with a number of centrifugations. These preliminary steps work towards developing an effective method for identifying and amplifying bacteriophage. The application of bacteriophage as an alternative treatment to Paenibacillus larvae bacteria, the causative agent of American Foulbrood, could then be explored in the near future with the ultimate goal of using bacteriophage in the hive.

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