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稀有 Lelliottia nimipressuralis 菌株 MEZLN61 和两株携带移动多粘菌素耐药基因 mcr-9 的 Enterobacter kobei 菌株 MEZEK193 和 MEZEK194 的基因组草图序列,这些菌株均从南非废水中分离得到。

Draft genome sequences of rare Lelliottia nimipressuralis strain MEZLN61 and two Enterobacter kobei strains MEZEK193 and MEZEK194 carrying mobile colistin resistance gene mcr-9 isolated from wastewater in South Africa.

机构信息

Veterinary Medicine and Food Security Research Group, Medical Laboratory Sciences Program, Faculty of Health Sciences, Abu Dhabi Women's Campus, Higher Colleges of Technology, Abu Dhabi 41012, UAE.

Institute of Hygiene and Environmental Medicine, Justus Liebig University Giessen, Giessen, Germany; German Center for Infection Research, Site Giessen-Marburg-Langen, Giessen, Germany; Hessian University Competence Center for Hospital Hygiene, Justus Liebig University Giessen, Giessen, Germany.

出版信息

J Glob Antimicrob Resist. 2023 Jun;33:231-237. doi: 10.1016/j.jgar.2023.03.007. Epub 2023 Mar 21.

Abstract

OBJECTIVES

Antimicrobial-resistant bacteria of the order Enterobacterales are emerging threats to global public and animal health, leading to morbidity and mortality. The emergence of antimicrobial-resistant, livestock-associated pathogens is a great public health concern. The genera Enterobacter and Lelliottia are ubiquitous, facultatively anaerobic, motile, non-spore-forming, rod-shaped Gram-negative bacteria belonging to the Enterobacteriaceae family and include pathogens of public health importance. Here, we report the first draft genome sequences of a rare Lelliottia nimipressuralis strain MEZLN61 and two Enterobacter kobei strains MEZEK193 and MEZEK194 in Africa.

METHODS

The bacteria were isolated from environmental wastewater samples. Bacteria were cultured on nutrient agar, and the pure cultures were subjected to whole-genome sequencing. Genomic DNA was sequenced using an Illumina MiSeq platform. Generated reads were trimmed and subjected to de novo assembly. The assembled contigs were analysed for virulence genes, antimicrobial resistance genes, and extra-chromosomal plasmids, and multilocus sequence typing was performed. To compare the sequenced strains with other, previously sequenced E. kobei and L. nimipressuralis strains, available raw read sequences were downloaded, and all sequence files were treated identically to generate core genome bootstrapped maximum likelihood phylogenetic trees.

RESULTS

Whole-genome sequencing analyses identified strain MEZLN61 as L. nimipressuralis and strains MEZEK193 and MEZEK194 as E. kobei. MEZEK193 and MEZEK194 carried genes encoding resistance to fosfomycin (fosA), beta-lactam antibiotics (bla), and colistin (mcr-9). Additionally, MEZEK193 harboured nine different virulence genes, while MEZEK194 harboured eleven different virulence genes. The phenotypic analysis showed that L. nimipressuralis strain MEZLN61 was susceptible to colistin (2 μg/mL), while E. kobei MEZEK193 (64 μg/mL) and MEZEK194 (32 μg/mL) were resistant to colistin.

CONCLUSION

The genome sequences of strains L. nimipressuralis MEZLN6, E. kobei MEZEK193, and E. kobei MEZEK194 will serve as a reference point for molecular epidemiological studies of L. nimipressuralis and E. kobei in Africa. In addition, this study provides an in-depth analysis of the genomic structure and offers important information that helps clarify the pathogenesis and antimicrobial resistance of L. nimipressuralis and E. kobei. The detection of mcr-9, which is associated with very low-level colistin resistance in Enterobacter species, is alarming and may indicate the undetected dissemination of mcr genes in bacteria of the order Enterobacterales. Continuous monitoring and surveillance of the prevalence of mcr genes and their associated phenotypic changes in clinically important pathogens and environmentally associated bacteria is necessary to control and prevent the spread of colistin resistance.

摘要

目的

肠杆菌目(Enterobacterales)的抗微生物耐药细菌对全球公共卫生和动物健康构成新兴威胁,导致发病率和死亡率上升。与牲畜相关的抗微生物耐药病原体的出现是一个重大的公共卫生关注点。肠杆菌属和莱氏利斯特氏菌属是普遍存在的兼性厌氧、能动、非孢子形成、杆状革兰氏阴性细菌,属于肠杆菌科,包括具有重要公共卫生意义的病原体。在这里,我们报告了在非洲首次发现的稀有莱氏利斯特氏菌(Lelliottia nimipressuralis)菌株 MEZLN61 和 2 株肠杆菌科(Enterobacter)的科氏肠杆菌(E. kobei)菌株 MEZEK193 和 MEZEK194 的完整基因组序列。

方法

从环境废水样本中分离细菌。在营养琼脂上培养细菌,然后对纯培养物进行全基因组测序。使用 Illumina MiSeq 平台对基因组 DNA 进行测序。生成的读数经过修剪并进行从头组装。对组装的连续体进行毒力基因、抗微生物耐药基因和染色体外质粒分析,并进行多位点序列分型。为了将测序菌株与其他先前测序的科氏肠杆菌和莱氏利斯特氏菌菌株进行比较,下载了可用的原始读取序列,并对所有序列文件进行了相同的处理,以生成核心基因组引导最大似然系统发育树。

结果

全基因组测序分析将菌株 MEZLN61 鉴定为莱氏利斯特氏菌,菌株 MEZEK193 和 MEZEK194 鉴定为科氏肠杆菌。MEZEK193 和 MEZEK194 携带编码对磷霉素(fosA)、β-内酰胺类抗生素(bla)和粘菌素(mcr-9)耐药的基因。此外,MEZEK193 携带 9 种不同的毒力基因,而 MEZEK194 携带 11 种不同的毒力基因。表型分析表明,莱氏利斯特氏菌(Lelliottia nimipressuralis)菌株 MEZLN61 对粘菌素(2 μg/mL)敏感,而科氏肠杆菌(E. kobei)MEZEK193(64 μg/mL)和 MEZEK194(32 μg/mL)对粘菌素耐药。

结论

莱氏利斯特氏菌 MEZLN6、科氏肠杆菌 MEZEK193 和科氏肠杆菌 MEZEK194 的基因组序列将为非洲莱氏利斯特氏菌和科氏肠杆菌的分子流行病学研究提供参考。此外,本研究对基因组结构进行了深入分析,并提供了重要信息,有助于阐明莱氏利斯特氏菌和科氏肠杆菌的发病机制和抗微生物耐药性。检测到与肠杆菌属中粘菌素低度耐药相关的 mcr-9 基因令人警惕,可能表明 mcr 基因在肠杆菌目细菌中的未检测到的传播。需要对临床上重要的病原体和与环境相关的细菌中 mcr 基因的流行情况及其相关表型变化进行持续监测和监测,以控制和预防粘菌素耐药性的传播。

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