Comparative proteomic analysis of virulent and rifampicin attenuated Flavobacterium psychrophilum

Flavobacterium psychrophilum is the aetiologic agent of bacterial coldwater disease and rainbow trout fry syndrome. In this study, we compared a wild-type strain (CSF 259-93) with a rifampicin-resistant strain and virulence-attenuated strain of F. psychrophilum (CSF 259-93B.17). The attenuated strai...

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Bibliographic Details
Published in:	Journal of fish diseases Vol. 35; no. 7; pp. 529 - 539
Main Authors:	Gliniewicz, K, Plant, K. P, LaPatra, S. E, LaFrentz, B. R, Cain, K, Snekvik, K. R, Call, D. R
Format:	Journal Article
Language:	English
Published:	Oxford, UK Blackwell Publishing Ltd 01-07-2012
Subjects:	Animals Antibodies, Bacterial - blood antigens antiserum Attenuation Bacterial Proteins - genetics Bacterial Proteins - immunology Bacterial Vaccines - immunology disease resistance Drug Resistance, Bacterial fish diseases Fish Diseases - immunology Fish Diseases - mortality Flavobacteriaceae Infections - immunology Flavobacteriaceae Infections - mortality Flavobacteriaceae Infections - veterinary Flavobacterium - genetics Flavobacterium - immunology Flavobacterium psychrophilum gene expression Gene Expression Regulation, Bacterial genes Immunization - veterinary immunoblotting mass spectrometry monoclonal antibodies mutation Oncorhynchus mykiss polyacrylamide gel electrophoresis protein synthesis Proteome proteomics recombinant proteins recombinant vaccines rifampicin Rifampin - metabolism subunit vaccine subunit vaccines Virulence - genetics
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Summary:	Flavobacterium psychrophilum is the aetiologic agent of bacterial coldwater disease and rainbow trout fry syndrome. In this study, we compared a wild-type strain (CSF 259-93) with a rifampicin-resistant strain and virulence-attenuated strain of F. psychrophilum (CSF 259-93B.17). The attenuated strain harboured a mutation in the rpoB gene consistent with resistance to rifampicin. Two-dimensional polyacrylamide gel electrophoresis (2DPAGE) and mass spectrometry demonstrated an altered proteome with eight proteins characteristic for the parent strain and six that were unique to the attenuated strain. Immunoblotting with a diagnostic monoclonal antibody (FL-43) identified a putative antigen (FP1493) that was subsequently cloned, expressed as a recombinant protein and confirmed as recognized by FL-43. 2D-PAGE, immunoblotting with rainbow trout, Oncorhynchus mykiss (Walbaum), convalescent antisera and mass spectrometry of bacterial whole-cell lysates revealed several uniquely expressed immunoreactive proteins including FP1493. An FP1493 recombinant subunit vaccine was tested, but did not provide protection against challenge with the CSF259-93 strain. While the exact mechanism responsible for altered protein synthesis and attenuation of CSF 259-93B.17 is still unknown, the differentially expressed immunoreactive proteins are a valuable resource to develop subunit vaccines and to identify proteins that are potentially involved in disease. Keywords: Attenuation, Flavobacterium psychrophilum, rifampicin, subunit vaccine.
Bibliography:	http://handle.nal.usda.gov/10113/55470 http://dx.doi.org/10.1111/j.1365-2761.2012.01378.x ArticleID:JFD1378 istex:6084A4A6015B7A35E58E4C2AF5206371375A41FD ark:/67375/WNG-2FRL6TP9-N ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1365-2761 0140-7775 1365-2761
DOI:	10.1111/j.1365-2761.2012.01378.x