Verhoeven Els, Abdellati Said, Nys Patrick, Laumen Jolein, De Baetselier Irith, Crucitti Tania, Kenyon Chris
Department of Clinical Sciences - STI unit, Institute of Tropical Medicine, Antwerp, Antwerp, 2000, Belgium.
Department of Clinical sciences - HIV/STI reference laboratory, Institute of Tropical Medicine, Antwerp, Antwerp, 2000, Belgium.
F1000Res. 2019 Apr 26;8:560. doi: 10.12688/f1000research.18861.2. eCollection 2019.
To obtain a detailed picture of the dynamics of antibiotic resistance development in , we built a morbidostat according to the protocol of Toprak ., adjusted to the specific characteristics required for the growth of . In this article we describe the adaptations, specifications and the difficulties we encountered during the construction and optimization of the NG morbidostat. As a proof of concept, we conducted a morbidostat experiment by increasing concentrations of azithromycin in response to bacterial growth. We started the experiment with two reference strains WHO-F and WHO-X. These strains were grown in 12 mL GC Broth supplemented with IsoVitaleX™ (1%) and vancomycin, colistin, nystatin, trimethoprim (VCNT) selective supplement for 30 days in a 6% CO environment at 36°C. Samples of the cultures were taken 2-3 times a week and minimal inhibitory concentrations (MICs) of azithromycin were determined using E-test. The initial MICs of WHO-F and WHO-X were 0.125 µg/mL and 0.25 µg/mL, respectively. In less than 30 days, we were able to induce high level azithromycin resistance in , with a 750 and 1000 fold increase in MIC for WHO-F and WHO-X, respectively.
为了详细了解抗生素耐药性产生的动态过程，我们按照托普拉克的方案构建了一台动态调整抑菌器，并根据其生长所需的特定特性进行了调整。在本文中，我们描述了在构建和优化NG动态调整抑菌器过程中的调整、规格以及遇到的困难。作为概念验证，我们通过根据细菌生长情况增加阿奇霉素浓度进行了动态调整抑菌器实验。我们用两种参考菌株WHO-F和WHO-X开始实验。这些菌株在补充了IsoVitaleX™（1%）以及万古霉素、黏菌素、制霉菌素、甲氧苄啶（VCNT）选择性补充剂的12 mL GC肉汤中，于36°C、6%二氧化碳环境下培养30天。每周采集2至3次培养物样本，并使用E-test法测定阿奇霉素的最低抑菌浓度（MIC）。WHO-F和WHO-X的初始MIC分别为0.125 µg/mL和0.25 µg/mL。在不到30天的时间里，我们成功诱导出了对阿奇霉素的高水平耐药性，WHO-F和WHO-X的MIC分别增加了750倍和1000倍。
