Gancz Ayala, Kondratyeva Kira, Cohen-Eli Dorit, Navon-Venezia Shiri
Molecular Biology Department, Faculty of Life Sciences, Ariel University, Ariel 40700, Israel.
The Miriam and Sheldon Adelson School of Medicine, Ariel University, Ariel 40700, Israel.
Microorganisms. 2021 May 10;9(5):1022. doi: 10.3390/microorganisms9051022.
The emergence of extended-spectrum β-lactamase (ESBL)-producing multidrug resistant causing community urinary tract infections (CA-UTI) in healthy women undermines effective treatment and poses a public health concern. We performed a comprehensive genomic analysis (Illumina and MinION) and virulence studies using nematodes to evaluate KpnU95, a -producing CA-UTI strain. Whole genome sequencing identified KpnU95 as sequence type 1412 and revealed the chromosomal and plasmid-encoding resistome, virulome and persistence features. KpnU95 possess a wide virulome and caused complete killing. The strain harbored a single novel 180.3Kb IncFIB(K) plasmid (pKpnU95), which encodes ten antibiotic resistance genes, including and alongside a wide persistome encoding heavy metal and UV resistance. Plasmid curing and reconstitution were used for loss and gain studies to evaluate its role on bacterial resistance, fitness and virulence. Plasmid curing abolished the ESBL phenotype, decreased ciprofloxacin MIC and improved bacterial fitness in artificial urine accompanied with enhanced copper tolerance, without affecting bacterial virulence. Meta-analysis supported the uniqueness of pKpnU95 and revealed plasmid-ST1412 lineage adaptation. Overall, our findings provide translational data on a CA-UTI ST1412 strain and demonstrates that ESBL-encoding plasmids play key roles in multidrug resistance and in bacterial fitness and persistence.
产超广谱β-内酰胺酶（ESBL）的多重耐药菌在健康女性中引起社区获得性尿路感染（CA-UTI），这削弱了有效治疗并引发了公共卫生问题。我们使用线虫进行了全面的基因组分析（Illumina和MinION）和毒力研究，以评估一株引起CA-UTI的产ESBL菌株KpnU95。全基因组测序确定KpnU95为序列型1412，并揭示了染色体和质粒编码的耐药组、毒力组和持续存在特征。KpnU95具有广泛的毒力组并导致完全杀灭。该菌株携带一个单一的新型180.3Kb IncFIB(K)质粒（pKpnU95），其编码十个抗生素抗性基因，包括blaCTX-M-15和blaTEM-1，同时还有一个广泛的持续存在组，编码重金属和紫外线抗性。质粒消除和重建用于缺失和获得研究，以评估其对细菌抗性、适应性和毒力的作用。质粒消除消除了ESBL表型，降低了环丙沙星的最低抑菌浓度（MIC），并提高了在人工尿液中的细菌适应性，同时增强了对铜的耐受性，而不影响细菌毒力。荟萃分析支持pKpnU95的独特性，并揭示了质粒-ST1412谱系适应性。总体而言，我们的研究结果提供了关于CA-UTI ST1412菌株的转化数据，并表明编码ESBL的质粒在多重耐药以及细菌适应性和持续存在中起关键作用。
