|Trott, Darren - USDA/ARS/NADC, AMES, IA|
|Wannemuehler, Mike - IOWA STATE UNIV, AMES, IA|
Submitted to: Research Workers in Animal Diseases Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: November 9, 1999
Publication Date: N/A
Technical Abstract: Brachyspira pilosicoli (Bp) is the etiologic agent of porcine intestinal spirochetosis (PIS), a colitis of growing pigs resulting in diarrhea and reduced growth rates. A unique feature of PIS is the attachment of large numbers of spirochetes by one cell end to the colonic epithelium. In order to identify Bp outer membrane proteins (OMPs) that may be involved in the pathogenesis of PIS we have developed a technique for gently removing Bp membranes by osmotic lysis. The membranes were separated by sucrose gradient ultracentrifugation into low density (LD) and high density (HD) fractions. Both membrane fractions were free of flagella and cytoplasmic proteins. Analysis of the fractions was performed using markers that were specific for the outer (cholesterol, lipooligosaccharide, 29 Kda OMP) and inner membrane (penicillin binding proteins and peptidoglycan). Outer membrane was present in both fractions, but only the LD fraction was significantly free of inner membrane proteins. The SDS-PAGE profiles of LD-membranes and HD-membranes differed considerably. The findings provide a basis for the further characterization of individual membrane proteins and elucidation of their role in PIS.