Myintzaw Peter, Pennone Vincenzo, McAuliffe Olivia, Begley Máire, Callanan Michael
Department of Biological Sciences, Munster Technological University, Bishopstown, T12 P928 Cork, Ireland.
Teagasc Food Research Centre, Moorepark, Fermoy, Co., P61 C996 Cork, Ireland.
Microorganisms. 2023 Jun 17;11(6):1603. doi: 10.3390/microorganisms11061603.
Precise classification of foodborne pathogen is a necessity in efficient foodborne disease surveillance, outbreak detection, and source tracking throughout the food chain. In this study, a total of 150 isolates from various food products, food processing environments, and clinical sources were investigated for variations in virulence, biofilm formation, and the presence of antimicrobial resistance genes based on their Whole-Genome Sequences. Clonal complex (CC) determination based on Multi-Locus Sequence Typing (MLST) revealed twenty-eight CC-types including eight isolates representing novel CC-types. The eight isolates comprising the novel CC-types share the majority of the known (cold and acid) stress tolerance genes and are all genetic lineage II, serogroup 1/2a-3a. Pan-genome-wide association analysis by Scoary using Fisher's exact test identified eleven genes specifically associated with clinical isolates. Screening for the presence of antimicrobial and virulence genes using the ABRicate tool uncovered variations in the presence of Pathogenicity Islands (LIPIs) and other known virulence genes. Specifically, the distributions of , , , , , LIPI-3, and genes across isolates were found to be significantly CC-dependent while the presence of , , , LIPI-3 was associated with clinical isolates specifically. In addition, Roary-derived phylogenetic grouping based on Antimicrobial-Resistant Genes (AMRs) revealed that the thiol transferase () gene was present in all lineage I isolates, and the presence of the lincomycin resistance ABC-F-type ribosomal protection protein () was also genetic-lineage-dependent. More importantly, the genes found to be specific to CC-type were consistent when a validation analysis was performed with fully assembled, high-quality complete genome sequences ( = 247) extracted from the National Centre for Biotechnology Information (NCBI) microbial genomes database. This work highlights the usefulness of MLST-based CC typing using the Whole-Genome Sequence as a tool in classifying isolates.
准确分类食源性病原体对于整个食物链中高效的食源性疾病监测、疫情检测和源头追踪至关重要。在本研究中,基于全基因组序列,对从各种食品、食品加工环境和临床来源获得的总共150株分离株进行了毒力、生物膜形成及抗菌抗性基因存在情况的变异研究。基于多位点序列分型(MLST)的克隆复合体(CC)测定揭示了28种CC型,其中包括8株代表新型CC型的分离株。构成新型CC型的8株分离株共享了大多数已知的(冷和酸)应激耐受基因,并且均为遗传谱系II,血清群1/2a - 3a。使用Fisher精确检验通过Scoary进行的全基因组关联分析确定了11个与临床分离株特异性相关的基因。使用ABRicate工具筛选抗菌和毒力基因的存在情况,发现致病性岛(LIPIs)和其他已知毒力基因的存在存在变异。具体而言,发现LIPI-1、LIPI-2、LIPI-4、LIPI-5、LIPI-6、LIPI-3和sopE2基因在分离株中的分布显著依赖于CC型,而LIPI-1、LIPI-2、LIPI-4、LIPI-3的存在则与临床分离株特异性相关。此外,基于抗微生物抗性基因(AMRs)的Roary衍生系统发育分组显示,硫醇转移酶(trxB)基因存在于所有谱系I分离株中,并且林可霉素抗性ABC - F型核糖体保护蛋白(lsaE)的存在也依赖于遗传谱系。更重要的是,当使用从国家生物技术信息中心(NCBI)微生物基因组数据库中提取的完全组装的高质量完整基因组序列(n = 247)进行验证分析时,发现特定于CC型的基因是一致的。这项工作突出了使用全基因组序列作为工具基于MLST的CC分型在分离株分类中的实用性。