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Title:   Author  item
                                            Brockmeier, Susanitem
                                            Register, Karenitem
                                            KUEHN, JOANNA - Iowa State Universityitem
                                            Nicholson, Tracyitem
                                            Loving, Crystalitem
                                            Shore, Sarahitem
                                            PHILLIPS, GREGORY - Iowa State University
  Submitted to: PLOS ONE
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 7/5/2014
                                    Publication  Date: 8/19/2014
                                    Publication URL: http://handle.nal.usda.gov/10113/61244Citation: Brockmeier, S.L., Register, K.B., Kuehn, J.S., Nicholson, T.L., Loving, C.L., Shore, S.M., Phillips, G.J. 2014. Virulence and draft genome sequence overview of multiple strains of the swine pathogen Haemophilus parasuis. PLoS ONEe. 9(8):e103787.Interpretive Summary: Haemophilus parasuis is a bacterium that causes Glässer's disease in swine, a disease characterized by chronic debilitation and often death that costs the swine industry millions in losses annually. However, not all strains of the bacterium cause disease. To date, little is known about genetic differences among H. parasuis strains and the genetic factors that contribute to its ability to cause disease. With a sensitive pig model of infection, we identified 10 strains of H. parasuis that varied in their ability to cause disease; from strains not causing any illness to strains that caused rapid onset of severe disease. We then determined the DNA genomic sequence of these 10 strains of H. parasuis. The DNA sequence can now be used to compare the different strains for identification of genes contributing to disease development. These data comprise the first publicly available genome sequence for this bacterium.
                                Technical Abstract: Haemophilus parasuis is the cause of Glässer’s disease in swine, which is characterized by systemic infection resulting in polyserositis, meningitis, and arthritis. Characterization of this animal disease is complicated by the enormous differences in the severity of disease caused by H. parasuis strains, ranging from lethal systemic disease to subclinical carriage. To identify differences in genotype that could account for virulence phenotypes, we established the virulence of, and performed whole genome sequence analysis on, 11 H. parasuis strains, 10 of which were sequenced as part of this study. Virulence was assessed by evaluating morbidity and mortality following intranasal challenge of caesarian-derived, colostrum-deprived (CDCD) pigs. Genomic DNA from strains Nagasaki, 12939, SW140, 29755, MN-H, 84-15995, SW114, H465, D74, and 174 was used to generate Illumina paired-end libraries for genomic sequencing and de novo assembly. H. parasuis strains Nagasaki, 12939, SH0165, SW140, 29755, and MN-H exhibited a high level of virulence. Despite minor distinctions among these groups, all pigs challenged with these strains developed clinical signs consistent with Glässer’s disease between 1-7 days post-challenge. H. parasuis strains 84-15995 and SW114 were moderately virulent, in that approximately half of the pigs infected with each developed Glässer’s disease. H. parasuis strains H465, D74, and 174 were minimally virulent or avirulent in the CDCD pig model. Comparative genomic analysis among strains identified several noteworthy differences in coding regions. These coding regions include predicted outer membrane, metabolism, and pilin or adhesin related genes, some of which likely contributed to the differences in virulence and systemic disease observed following challenge. These data will be useful for identifying H. parasuis virulence factors and vaccine targets.


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