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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #337339

Research Project: Non-Antibiotic Strategies to Control Priority Bacterial Infections in Swine

Location: Virus and Prion Research

Title: The quest for cross protective factors of Haemophilus parasuis using 2-D gel electrophoresis

item EBERLE, K - Orise Fellow
item Nally, Jarlath
item Lippolis, John
item Loving, Crystal
item HAU, S - Iowa State University
item Nicholson, Tracy
item Brockmeier, Susan

Submitted to: American Society for Microbiology General Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 2/27/2017
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: In swine, Haemophilus parasuis (H. parasuis) infection causes polyserositis, arthritis, and meningitis. A range of virulent to nonvirulent strains exists between and within the 15 serovars. Because of this, the pathogenicity and subsequent protection from H. parasuis disease has yet to be elucidated. Bacterins made from two H. parasuis serovar 5 (SV5) strains were used to examine both homologous and heterologous protective antibodies produced during infection and therefore identify possible cross protective factors. Pigs were vaccinated with a H. parasuis SV5 bacterin made from either 265 or Nagasaki. Though both bacterins protected against challenge with the homologous strain, a difference in protection was observed against a heterologous challenge with 12939, a SV1 strain. That is, pigs given the Nagasaki bacterin were not protected against the heterologous 12939 challenge while those vaccinated with the 265 bacterin were all protected. ELISA antibody titer using whole bacterial sonicate revealed similar antibody levels for both vaccination methods to homologous and heterologous H. parasuis. Therefore, a higher level of antibody production in those animals given the 265 bacterin was not the source of protection against 12939. Because the outer membrane proteins (OMPs) are those predicted to be involved in virulence and antibody protection, we used a Triton X-114 enrichment method to isolate the OMPs of H. parasuis. A western blot was probed using antiserum from either protected (265 bacterin) or nonprotected (Nagasaki bacterin) pigs against the OMPs of H. parasuis 12939. Stark differences in the possible protective antibodies produced during vaccination were observed. Through 2-D gel electrophoresis, we further separated proteins of interest and identified them by mass spectrometry. These bacterial OMPs are the targets of protective antibodies and potential cross protective factors.