Abstract
The aim of this study was to identify the immunodominant outer membrane proteins (OMPs) of Fusobacterium necrophorum from sheep affected with severe foot-rot. The OMP profile of ovine strains of F. necrophorum has not been well studied. We analyzed the OMP profile of the most frequent lktA variant JKS-F3 of F. necrophorum associated with severe ovine foot-rot with lesion score 4 in order to identify its major immunodominant OMPs. Electrophoretic separations of extracted OMPs showed a number of spots in two-dimensional electrophoretic gels. Two immunoreactive proteins of size around 43 kDa were identified through western blotting using hyperimmune sera raised in rabbits. These two immunogenic OMPs were analyzed by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF/MS) which revealed that these two OMPs of lktA variant JKS-F3 of F. necrophorum showed 46 and 42 percent protein sequence coverage and scores of 125 and 114, respectively, with the reported 43 kDa outer membrane protein of F. necrophorum strain H05, a putative porin having properties similar to pore-forming proteins of anaerobic Gram-negative bacteria. These identified immunogenic OMPs will contribute to our understanding of the pathogenic role played by this organism in ovine foot-rot and could be exploited to devise an effective control strategy through development of an OMP-based recombinant vaccine to mitigate foot-rot in sheep and goats.
Highlights
Fusobacterium necrophorum is a Gram-negative, rodshaped, non-spore-forming anaerobic bacterium that is associated with a variety of severe and fatal diseases in cattles, sheep and humans
Given the lack of information on the outer membrane proteins (OMPs) profile of ovine strains of F. necrophorum, we identified and analyzed the OMPs of most predominant lktA variant JKS-F3 of F. necrophorum associated with severe foot-rot lesions in sheep by MALDI-ToF mass spectrometry
Like other Gramnegative bacteria, outer membrane proteins (OMPs) of F. necrophorum are important in facilitating attachment and is the first bacterial component to come in contact with the host cell surface and play a vital role in establishment of infection and virulence [19]
Summary
Fusobacterium necrophorum is a Gram-negative, rodshaped, non-spore-forming anaerobic bacterium that is associated with a variety of severe and fatal diseases in cattles, sheep and humans. F. necrophorum cause Lemierre syndrome, which begins as pharyngitis and rapidly progresses to septic thrombophlebitis of the jugular vein [1]. Fnf is mostly implicated in human infection while Fnn being the principal animal pathogen mainly responsible for hepatic abscesses in beef cattle [2–4] and severe foot-rot in sheep and goats along with. These anaerobes are more commonly detected in footrot affected than in healthy feet of sheep; both need to be considered when managing a footrot outbreak [7]. F. necrophorum is an important pathogen contributing to the severity and duration of severe footrot (SFR) in sheep and goats and is often found in association with D. nodosus in infected feet with lesion score of mostly 4 [5, 6, 10]
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