Outer membrane protein profiles of Haemophilus pleuropneumoniae

Abstract

Outer membrane protein profiles of Haemophilus pleuropneumoniae were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cells were disrupted by sonication, and outer membrane-enriched fractions were prepared by differential centrifugation and selective solubilization of the inner membrane with sodium N-lauroyl sarcosinate. Colony type, growth medium, time of harvest, and in vitro or in vivo passage had no appreciable effect on the protein profiles of the strains examined. Seven patterns were distinguished among the reference strains of the nine capsular serotypes. These patterns were based on the mobility of the major outer membrane proteins migrating in the 39,000to 44,000-molecular-weight region of the gel, a 16K to 16.5K protein, and a heat-modifiable 29K protein. Strains of serotypes 1 and 9 had identical outer membrane protein profiles, as did strains of serotypes 2 and 6. The reference strains of the remaining five serotypes each had a distinct pattern. The outer membrane protein proffles of 95 field isolates belonging to serotypes 1, 5, 7, and 9 from swine in the midwestern United States were determined and compared with the reference patterns. The results indicate that the population of H. pleuropneumoniae is clonal, with three predominant clones distinguished by both serotype and outer membrane protein profile responsible for the majority of H. pleuropneumoniae disease occurring in swine in the United States.

Efficacy of a cell extract from Actinobacillus (Haemophilus) pleuropneumoniae serotype 1 against disease in swine

“…SDS-PAGE analysis of the OMP preparations from serotypes 1, 5, and 7 were consistent with those reported by Rapp et al (32) and Rycroft and Taylor (37). We have confirmed previous findings (33) that A. pleuropneumoniae OMP provided some protection against pleuropneumonia in pigs; however, it appeared to be less effective than the CE.…”

Serological responses to two serovar‐independent ELISA antigens of Actinobacillus pleuropneumoniae in Australian commercial pig herds

“…Additional serovar‐independent, outer membrane protein (OMP) antigens are present in A. pleuropneumoniae . Major OMPs of varying seroreactivity and approximately 39 to 44 kDa in size are shared among several serovars, 12,13 although studies at our laboratory have demonstrated an immunodominant, surface accessible 39‐kDa OMP is seroreactive and shared among all A. pleuropneumoniae serovars 14–17 . Because similar OMPs may exist among related pig pathogens, including Pasteurella multocida 18,19 and Haemophilus parasuis , 20 and antisera to these may be cross‐reactive with a 40‐kDa OMP of A. pleuropneumoniae , 20 evaluation of the diagnostic test specificity of the 39‐kDa ELISA antigen was considered important.…”

Influence of Photosensitizer Solvent on the Mechanisms of Photoactivated Killing of Enterococcus faecalis

“…Equal number of E. faecalis cells (10 9 cells mL )1 ) subjected to PDT as in previous experiments were collected by centrifugation. The outer membrane proteins were isolated as detailed elsewhere (20). Briefly, the cells collected were suspended in HEPES buffer (pH 7.4) and cells were disrupted with a sonicator at 15 kHz output (Sonoplus, Bandelin, Germany).…”