There are several methods for cultivation of Paenibacillus larvae, the causative agent of American foulbrood (AFB) in honey bees. Protocols for detection of sub-clinical levels of the bacterium from honey and bee samples include heat treatment of samples. The main objective of this study was to investigate if there is variability in temperature resistance among P. larvae genotypes, potentially leading to biased diagnose and disease monitoring. The variation in germination and proliferation ability among type collection (N=4) and field isolates (N=4) of P. larvae representing four different genotypes was investigated. Results demonstrate a significant variability between P. larvae genotypes in germination rate on solid media as well as in endospore resistance to heat treatment and storage. It is concluded that strains of different genotypes should be included in evaluation of standard laboratory protocols for cultivation of P. larvae to avoid bias in disease monitoring and quantification of the pathogen.