American foulbrood (AFB) is a devastating brood disease of the honey bee caused by the spore-forming, Gram-positive rod-shaped bacterium Paenibacillus larvae. AFB is one of the bee diseases listed in the OIE (Office International des Epizooties – the World Organization for Animal Health) Terrestrial Animal Health Code (2011) and member countries and territories are obliged to report its occurrence. In 2006, a draft of the P. larvae genome was published at an estimated 5-6x coverage (Qin et al., 2006). Last year, this coverage was further extended and the genome sequence was further annotated with a combination of bioinformatics and proteomics (Chan et al., 2011). These efforts will certainly help to usher in the next level of research for this economically important pathogen, ultimately allowing us to better understand the intimate relationship between the pathogen and its host. More generally, the honey bee / AFB system provides a wealth of opportunities and tools for addressing basic questions regarding microbe-microbe interactions, host immunity, strain virulence, and horizontal transmission, among others.
In the present paper, selected techniques and protocols in American foulbrood research are provided, mostly in a recipe-like format that permits easy implementation in the laboratory. The different topics that are covered include: working with Paenibacillus larvae, basic microbiological techniques, experimental infection and “’omics” and other sophisticated techniques. Thus, the chapter covers a broad set of technical information going from biosafety measures to guarantee the safe handling of this pathogen to the expression of heterologous proteins in P. larvae. Techniques exclusively related to the diagnosis of AFB are not included as they have been reviewed elsewhere (de Graaf et al., 2006a; OIE, 2008).