Brockmeier Susan L, Register Karen B, Kuehn Joanna S, Nicholson Tracy L, Loving Crystal L, Bayles Darrell O, Shore Sarah M, Phillips Gregory J
Virus and Prion Diseases Research Unit, National Animal Disease Center, Agricultural Research Service, USDA, Ames, Iowa, United States of America.
Ruminant Diseases and Immunology Research Unit, National Animal Disease Center, Agricultural Research Service, USDA, Ames, Iowa, United States of America.
PLoS One. 2014 Aug 19;9(8):e103787. doi: 10.1371/journal.pone.0103787. eCollection 2014.
Haemophilus parasuis is the cause of Glässer's disease in swine, which is characterized by systemic infection resulting in polyserositis, meningitis, and arthritis. Investigation 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. Virulence was assessed by evaluating morbidity and mortality following intranasal challenge of Caesarean-derived, colostrum-deprived (CDCD) pigs. Genomic DNA from strains Nagasaki (serotype 5), 12939 (serotype 1), SW140 (serotype 2), 29755 (serotype 5), MN-H (serotype 13), 84-15995 (serotype 15), SW114 (serotype 3), H465 (serotype 11), D74 (serotype 9), and 174 (serotype 7) was used to generate Illumina paired-end libraries for genomic sequencing and de novo assembly. H. parasuis strains Nagasaki, 12939, SH0165 (serotype 5), SW140, 29755, and MN-H exhibited a high level of virulence. Despite minor differences in expression of disease 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.
副猪嗜血杆菌是猪格氏病的病原体,该病的特征是全身感染,可导致多浆膜炎、脑膜炎和关节炎。由于副猪嗜血杆菌菌株引起的疾病严重程度差异巨大,从致死性全身疾病到亚临床携带状态,对这种动物疾病的研究变得复杂。为了确定可能导致毒力表型差异的基因型差异,我们对11株副猪嗜血杆菌菌株进行了毒力测定,并进行了全基因组序列分析。通过评估剖腹产、初乳缺乏(CDCD)仔猪经鼻攻毒后的发病率和死亡率来评估毒力。使用来自长崎(血清型5)、12939(血清型1)、SW140(血清型2)、29755(血清型5)、MN-H(血清型13)、84-15995(血清型15)、SW114(血清型3)、H465(血清型11)、D74(血清型9)和174(血清型7)菌株的基因组DNA来构建Illumina双末端文库,用于基因组测序和从头组装。副猪嗜血杆菌长崎株、12939株、SH0165(血清型5)、SW140株、29755株和MN-H株表现出高毒力。尽管这些组之间在疾病表现上存在微小差异,但所有用这些菌株攻毒的仔猪在攻毒后1至7天内都出现了与格氏病一致的临床症状。副猪嗜血杆菌84-15995株和SW114株为中等毒力,因为感染这两种菌株的仔猪中约有一半患上了格氏病。副猪嗜血杆菌H465株、D74株和174株在CDCD仔猪模型中为低毒力或无毒力。菌株间的比较基因组分析在编码区发现了几个值得注意的差异。这些编码区包括预测的外膜、代谢以及菌毛或黏附素相关基因,其中一些可能导致了攻毒后观察到的毒力和全身疾病差异。这些数据将有助于确定副猪嗜血杆菌的毒力因子和疫苗靶点。