Domingues Sara, Lima Tiago, Escobar Corentin, Plantade Julie, Charpentier Xavier, da Silva Gabriela Jorge
Faculty of Pharmacy, University of Coimbra, Coimbra, Portugal.
CNC-UC - Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal.
Eur J Clin Microbiol Infect Dis. 2025 Jun;44(6):1417-1424. doi: 10.1007/s10096-025-05113-9. Epub 2025 Mar 28.
Antimicrobial resistance poses a significant global health challenge, contributing to a lack of effective therapeutic agents, especially against Gram-negative bacteria. Resistance dissemination is accelerated by horizontal gene transfer (HGT) mechanisms. The extended-spectrum beta lactamases CTX-M confer resistance to several beta-lactams, are usually embedded into plasmids and thought to be mainly disseminated by conjugation. However, an increasing number of isolates carry these enzyme-encoding genes in the chromosome, suggesting that they can spread by other means of HGT. In this study, we aimed to test the involvement of natural transformation in the chromosomal acquisition of a bla gene.
Natural transformation assays were performed during motility on wet surfaces. Acquisition of foreign DNA by transformants was screened by antimicrobial susceptibility testing, polymerase-chain reaction (PCR) and whole genome sequencing (WGS).
Acinetobacter baumannii A118, a naturally competent clinical strain, was transformed with naked DNA from Salmonella enterica serovar Typhimurium Sal25, which was isolated from swine meat. The transformation occurred at low frequency (2.7 × 10 ± 2.04 × 10 transformants per recipient) and bla was acquired in one transformant, which was named ACI. WGS of the transformant revealed the acquisition of the bla as part of a ca. 36 Kb DNA fragment through an ISEc9-mediated transposition event; various mobile genetic elements and other resistance genes were co-transferred. The bla gene was subsequently transferred within A. baumannii at a higher frequency (1.8 × 10 ± 2.49 × 10 transformants per recipient).
Our results highlight the importance of natural transformation events in the dissemination of antimicrobial resistance genes and mobile genetic elements between and within species.
抗菌药物耐药性对全球健康构成重大挑战,导致缺乏有效的治疗药物,尤其是针对革兰氏阴性菌的药物。水平基因转移(HGT)机制加速了耐药性的传播。超广谱β-内酰胺酶CTX-M对多种β-内酰胺类药物具有耐药性,通常嵌入质粒中,被认为主要通过接合作用传播。然而,越来越多的分离株在染色体上携带这些酶编码基因,这表明它们可以通过其他HGT方式传播。在本研究中,我们旨在测试自然转化在bla基因染色体获得过程中的作用。
在潮湿表面的运动过程中进行自然转化试验。通过抗菌药物敏感性试验、聚合酶链反应(PCR)和全基因组测序(WGS)筛选转化体对外源DNA的获得情况。
鲍曼不动杆菌A118是一种天然感受态临床菌株,用从猪肉中分离的鼠伤寒沙门氏菌Sal25的裸DNA进行转化。转化发生频率较低(每个受体有2.7×10±2.04×10个转化体),在一个转化体中获得了bla,该转化体命名为ACI。对该转化体的WGS分析显示,通过ISEc9介导的转座事件,bla作为约36 Kb DNA片段的一部分被获得;各种移动遗传元件和其他耐药基因也被共同转移。随后,bla基因在鲍曼不动杆菌内以更高的频率转移(每个受体有1.8×10±2.49×10个转化体)。
我们的结果突出了自然转化事件在抗菌耐药基因和移动遗传元件在种间和种内传播中的重要性。