Pedreira Adrián, Vázquez José A, Romanenko Andrey, García Míriam R
Biosystems and Bioprocess Engineering Group (Bio2Eng), Spanish National Research Council (IIM-CSIC), Rúa Eduardo Cabello 6, 36208 Vigo, Spain.
Group of Recycling and Valorization of Waste Materials (REVAL), Spanish National Research Council (IIM-CSIC), Rúa Eduardo Cabello 6, 36208 Vigo, Spain.
Bioengineering (Basel). 2024 Aug 10;11(8):815. doi: 10.3390/bioengineering11080815.
During adaptive laboratory evolution experiments, any unexpected interruption in data monitoring or control could lead to the loss of valuable experimental data and compromise the integrity of the entire experiment. Most homemade mini-bioreactors are built employing microcontrollers such as Arduino. Although affordable, these platforms lack the robustness of the programmable logic controller (PLC), which enhances the safety and robustness of the control process. Here, we describe the design and validation of a PLC-controlled morbidostat, an innovative automated continuous-culture mini-bioreactor specifically created to study the evolutionary pathways to drug resistance in microorganisms. This morbidostat includes several improvements, both at the hardware and software level, for better online monitoring and a more robust operation. The device was validated employing , exploring its adaptive evolution in the presence of didecyldimethylammonium chloride (DDAC), a quaternary ammonium compound widely used for its antimicrobial properties. was subjected to increasing concentrations of DDAC over 3 days. Our results demonstrated a significant increase in DDAC susceptibility, with evolved populations exhibiting substantial changes in their growth after exposure.
在适应性实验室进化实验期间，数据监测或控制中的任何意外中断都可能导致宝贵实验数据的丢失，并损害整个实验的完整性。大多数自制的微型生物反应器是使用诸如Arduino之类的微控制器构建的。尽管价格实惠，但这些平台缺乏可编程逻辑控制器（PLC）的稳健性，而PLC可增强控制过程的安全性和稳健性。在此，我们描述了一种由PLC控制的病态恒化器的设计与验证，这是一种专门为研究微生物耐药性进化途径而创建的创新型自动化连续培养微型生物反应器。这种病态恒化器在硬件和软件层面都有多项改进，以实现更好的在线监测和更稳健的运行。该设备通过在存在二癸基二甲基氯化铵（DDAC，一种因其抗菌特性而广泛使用的季铵化合物）的情况下探索其适应性进化来进行验证。在3天内使 暴露于不断增加浓度的DDAC中。我们的结果表明，DDAC敏感性显著增加，进化后的群体在暴露后其生长表现出实质性变化。
