Intra-Specific Diversity within Pasteurella Trehalosi based on Variation of Capsular Polysaccharide, Lipopolysaccharide and Outer-Membrane Proteinsx

Abstract

Intra-specific diversity within Pasteurella trehalosi was investigated by analysis of variation of capsular polysaccharide, and lipopolysaccharide (LPS) and outer-membrane protein (OMP) profiles. Sixty isolates of P. trehalosi from diverse geographical locations within the UK, were examined. Capsular polysaccharide serotypes were determined by indirect haemagglutination assay; LPS and OMP profiles were compared by SDS-PAGE analysis. Capsular serotyping identified three isolates of serotype T3, 18 isolates each of serotypes T4, T10 and T15, and three untypable (UT) isolates. Analysis of LPS and OMP profiles identified six smooth LPS types and four OMP types among the 60 isolates. Forty-five (75%) of the isolates belonged to a single OMP type whereas 52 (87%) of the isolates possessed one of three LPS types. Each typing method, by itself, was not very discriminating but when the data from the three methods were combined, the 60 isolates could be separated into 14 distinct subgroups containing from one to 16 isolates as follows: serotype T3, two subgroups; serotype T4, four subgroups; serotype T10, two subgroups; serotype T15, five subgroups; UT isolates, one subgroup. Certain subgroups were associated with only one serotype whereas other subgroups were common to two or more serotypes. The subgroupings were capable of differentiating between isolates of the same serotype from the same and different geographical origins. Based on their LPS and OMP profiles, isolates of serotypes T4 and T15 were more closely related to each other than to isolates of serotype T10; serotype T4 and T15 isolates were also more heterogeneous than those of serotype T10. Certain isolates of serotype T10, recovered from a wide geographical area, were characterized by the possession of a unique capsule/LPS/OMP combination and represented a single clonal group which was responsible for a large proportion (31%) of recent disease outbreaks. Overall, a combination of capsular serotyping, and LPS and OMP typing, was found to be extremely useful for assessing diversity within P. trehalosi and should be of value for epidemiological and virulence studies.