Department of Veterinary Diagnostic and Production Animal Medicine, Iowa State University, Ames, IA, USA.
Groupe de Recherche Sur Les Maladies Infectieuses du Porc, Faculté de Médecine Vétérinaire, Université de Montréal, Montréal, Canada.
BMC Vet Res. 2023 Aug 28;19(1):135. doi: 10.1186/s12917-023-03698-x.
Glaesserella parasuis is the causative agent of Glässer's disease in pigs. Serotyping is the most common method used to type G. parasuis isolates. However, the high number of non-typables (NT) and low discriminatory power make serotyping problematic. In this study, 218 field clinical isolates and 15 G. parasuis reference strains were whole-genome sequenced (WGS). Multilocus sequence types (MLST), serotypes, core-genome phylogeny, antimicrobial resistance (AMR) genes, and putative virulence gene information was extracted.
In silico WGS serotyping identified 11 of 15 serotypes. The most frequently detected serotypes were 7, 13, 4, and 2. MLST identified 72 sequence types (STs), of which 66 were novel. The most predominant ST was ST454. Core-genome phylogeny depicted 3 primary lineages (LI, LII, and LIII), with LIIIA sublineage isolates lacking all vtaA genes, based on the structure of the phylogenetic tree and the number of virulence genes. At least one group 1 vtaA virulence genes were observed in most isolates (97.2%), except for serotype 8 (ST299 and ST406), 15 (ST408 and ST552) and NT (ST448). A few group 1 vtaA genes were significantly associated with certain serotypes or STs. The putative virulence gene lsgB, was detected in 8.3% of the isolates which were predominantly of serotype 5/12. While most isolates carried the bcr, ksgA, and bacA genes, the following antimicrobial resistant genes were detected in lower frequency; blaZ (6.9%), tetM (3.7%), spc (3.7%), tetB (2.8%), bla-ROB-1 (1.8%), ermA (1.8%), strA (1.4%), qnrB (0.5%), and aph3''Ia (0.5%). CONCLUSION: This study showed the use of WGS to type G. parasuis isolates and can be considered an alternative to the more labor-intensive and traditional serotyping and standard MLST. Core-genome phylogeny provided the best strain discrimination. These findings will lead to a better understanding of the molecular epidemiology and virulence in G. parasuis that can be applied to the future development of diagnostic tools, autogenous vaccines, evaluation of antibiotic use, prevention, and disease control.
猪副嗜血杆菌(Glaesserella parasuis)是导致格拉泽氏病的病原体。血清型鉴定是鉴定 G. parasuis 分离株最常用的方法。然而,非定型菌株(NT)数量较多且鉴别力较低,使得血清型鉴定存在问题。本研究对 218 株临床分离株和 15 株 G. parasuis 参考株进行了全基因组测序(WGS)。提取了多位点序列分型(MLST)、血清型、核心基因组系统发育、抗生素耐药(AMR)基因和潜在毒力基因信息。
基于 WGS 的血清型鉴定可鉴定出 15 种血清型中的 11 种。最常见的血清型为 7、13、4 和 2。MLST 鉴定出 72 种序列型(ST),其中 66 种为新 ST。最主要的 ST 为 ST454。核心基因组系统发育描绘了 3 个主要谱系(LI、LII 和 LIII),LIIIA 亚谱系分离株缺乏所有 vtaA 基因,这基于系统发育树的结构和毒力基因的数量。除血清型 8(ST299 和 ST406)、15(ST408 和 ST552)和 NT(ST448)外,大多数分离株(97.2%)均至少携带一组 1 vtaA 毒力基因。少数组 1 vtaA 基因与某些血清型或 ST 显著相关。在 8.3%的分离株中检测到了假定的毒力基因 lsgB,这些分离株主要为血清型 5/12。大多数分离株携带 bcr、ksgA 和 bacA 基因,但检测到较低频率的以下抗生素耐药基因:blaZ(6.9%)、tetM(3.7%)、spc(3.7%)、tetB(2.8%)、bla-ROB-1(1.8%)、ermA(1.8%)、strA(1.4%)、qnrB(0.5%)和 aph3''Ia(0.5%)。
本研究表明,使用 WGS 对 G. parasuis 分离株进行分型,可以作为更耗时和传统的血清型鉴定和标准 MLST 的替代方法。核心基因组系统发育提供了最佳的菌株鉴别力。这些发现将有助于更好地了解 G. parasuis 的分子流行病学和毒力,可应用于未来诊断工具的开发、同源疫苗的开发、抗生素使用的评估、预防和疾病控制。