1887

Abstract

American foulbrood is a bacterial disease of worldwide distribution that affects larvae of the honeybee The causative agent is the Gram-positive, spore-forming bacterium . Several authors have proposed that secretes metalloproteases that are involved in the larval degradation that occurs after infection. The aim of the present work was to evaluate the production of a metalloprotease by during larval infection. First, the complete gene encoding a metalloprotease was identified in the genome and its distribution was evaluated by PCR in a collection of isolates from different geographical regions. Then, the complete gene was amplified, cloned and overexpressed, and the recombinant metalloprotease was purified and used to generate anti-metalloprotease antibodies. Metalloprotease production was evaluated by immunofluorescence and fluorescence hybridization. The gene encoding a metalloprotease was widely distributed in isolates from different geographical origins in Uruguay and Argentina. Metalloprotease was detected inside vegetative cells, on the surface of spores and secreted to the external growth medium. Its production was also confirmed during the infection of honeybee larvae. This protein was able to hydrolyse milk proteins as described for suggesting that could be involved in larval degradation. This work contributes to the knowledge of the pathogenicity mechanisms of a bacterium of great economic significance and is one step in the characterization of potential virulence factors.

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2011-05-01
2019-10-21
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