Multidrug-resistant Klebsiella pneumoniae and Klebsiella variicola isolated from patients in Portuguese hospitals: Genomic and phenotypic characterization

Joana Castro , Daniela Araújo , Hugo Oliveira , Liliana Fernandes , Ricardo Oliveira , Erick Brinks , Gyu-Sung Cho , Charles Franz , Maria José Saavedra , Sónia Silva , Carina Almeida
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Abstract

The emergence of antibiotic-resistant Klebsiella spp. has become a huge problem in clinical settings. In this study, we aimed to characterize the molecular and phenotypic features of 21 MDR Klebsiella isolates (n = 19 of K. pneumoniae, and n = 2 of K. variicola) isolated from patients admitted to two central hospitals in northern Portugal by using whole-genome sequencing (WGS), followed by an in vitro assessment of biofilm formation and in vivo evaluation of the pathogenicity in a Galleria mellonella larval model. Our findings showed a high prevalence of O1/O2 serotypes (14/21; 67 %) among the isolates tested, which is consistent with previous reports from Portugal. In contrast, a wide variety of K locus serotypes was found, where ST15-KL19 (4/21; 19 %) associated to serotype O1/O2v2 was the dominant one. Within the O1/O2v2 serotype, a ST10-KL151 Klebsiella variicola (strain H97) harboured a high number of virulence genes. We also found statistical differences in the ability to produce biofilm biomass within the strains, with a ST280-KL23 K. pneumoniae outcompeting nine other strains. According to our results, the most prominent serotype able to cause the death of G. mellonella was the KL105-O1/O2v2. Our findings emphasized the importance of the conducting continuous molecular surveillance in order to reveal the key molecular features to be considered in the development of novel strategies to treat Klebsiella spp.-associated infections.
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从葡萄牙医院病人中分离出的耐多药肺炎克雷伯菌和变异克雷伯菌:基因组和表型特征
抗生素耐药克雷伯菌属的出现已成为临床环境中的一个巨大问题。在本研究中,我们通过全基因组测序(WGS)技术,对从葡萄牙北部两家中心医院收治的患者中分离出的 21 株 MDR 克雷伯氏菌(肺炎克雷伯菌 19 株,变异克雷伯菌 2 株)进行了分子和表型特征描述,随后在体外评估了生物膜的形成,并在体内评估了在黑嘴鸥幼虫模型中的致病性。我们的研究结果表明,在测试的分离株中,O1/O2 血清型的流行率很高(14/21;67%),这与葡萄牙以前的报告一致。相比之下,我们发现了多种 K 基因座血清型,其中与血清型 O1/O2v2 相关的 ST15-KL19 (4/21;19%)是最主要的血清型。在 O1/O2v2 血清型中,ST10-KL151变异克雷伯氏菌(菌株 H97)含有大量毒力基因。我们还发现,菌株产生生物膜生物量的能力存在统计学差异,ST280-KL23 肺炎克雷伯氏菌优于其他九种菌株。根据我们的研究结果,能导致 G. mellonella 死亡的最主要血清型是 KL105-O1/O2v2。我们的发现强调了进行持续分子监测的重要性,以揭示在开发治疗克雷伯氏菌属相关感染的新策略时应考虑的关键分子特征。
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