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Putative determinants of virulence in Melissococcus plutonius, the bacterial agent causing European foulbrood in honey bees.

ABSTRACT:

Melissococcus plutonius

is a bacterial pathogen that causes epidemic outbreaks of European foulbrood (EFB) in honey bee populations. The pathogenicity of a bacterium depends on its virulence, and understanding the mechanisms influencing virulence may allow for improved disease control and containment. Using a standardized in vitro assay, we demonstrate that virulence varies greatly among sixteen M. plutonius isolates from five European countries. Additionally, we explore the causes of this variation. In this study, virulence was independent of the multilocus sequence type of the tested pathogen, and was not affected by experimental co-infection with Paenibacillus alvei, a bacterium often associated with EFB outbreaks. Virulence in vitro was correlated with the growth dynamics of M. plutonius isolates in artificial medium, and with the presence of a plasmid carrying a gene coding for the putative toxin melissotoxin A. Our results suggest that some M. plutonius strains showed an increased virulence due to the acquisition of a toxin-carrying mobile genetic element. We discuss whether strains with increased virulence play a role in recent EFB outbreaks.

SUBMITTER: Grossar D

PROVIDER: S-EPMC7567439 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Putative determinants of virulence in <i>Melissococcus plutonius</i>, the bacterial agent causing European foulbrood in honey bees.

Grossar Daniela D Kilchenmann Verena V Forsgren Eva E Charrière Jean-Daniel JD Gauthier Laurent L Chapuisat Michel M Dietemann Vincent V

Virulence 20201201 1

<h4>Melissococcus plutonius</h4>is a bacterial pathogen that causes epidemic outbreaks of European foulbrood (EFB) in honey bee populations. The pathogenicity of a bacterium depends on its virulence, and understanding the mechanisms influencing virulence may allow for improved disease control and containment. Using a standardized <i>in vitro</i> assay, we demonstrate that virulence varies greatly among sixteen <i>M. plutonius</i> isolates from five European countries. Additionally, we explore th ...[more]

PMID: 32456539

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Project description:European foulbrood (EFB) is an important infectious disease of honeybee larvae, but its pathogenic mechanisms are still poorly understood. The causative agent, Melissococcus plutonius, is a fastidious organism, and microaerophilic to anaerobic conditions and the addition of potassium phosphate to culture media are required for growth. Although M. plutonius is believed to be remarkably homologous, in addition to M. plutonius isolates with typical cultural characteristics, M. plutonius-like organisms, with characteristics seemingly different from those of typical M. plutonius, have often been isolated from diseased larvae with clinical signs of EFB in Japan. Cultural and biochemical characterization of 14 M. plutonius and 19 M. plutonius-like strain/isolates revealed that, unlike typical M. plutonius strain/isolates, M. plutonius-like isolates were not fastidious, and the addition of potassium phosphate was not required for normal growth. Moreover, only M. plutonius-like isolates, but not typical M. plutonius strain/isolates, grew anaerobically on sodium phosphate-supplemented medium and aerobically on some potassium salt-supplemented media, were positive for ?-glucosidase activity, hydrolyzed esculin, and produced acid from L-arabinose, D-cellobiose, and salicin. Despite the phenotypic differences, 16S rRNA gene sequence analysis and DNA-DNA hybridization demonstrated that M. plutonius-like organisms were taxonomically identical to M. plutonius. However, by pulsed-field gel electrophoresis analysis, these typical and atypical (M. plutonius-like) isolates were separately grouped into two genetically distinct clusters. Although M. plutonius is known to lose virulence quickly when cultured artificially, experimental infection of representative isolates showed that atypical M. plutonius maintained the ability to cause EFB in honeybee larvae even after cultured in vitro in laboratory media. Because the rapid decrease of virulence in cultured M. plutonius was a major impediment to elucidation of the pathogenesis of EFB, atypical M. plutonius discovered in this study will be a breakthrough in EFB research.