Klopffer Lucie, Louvet Nicolas, Becker Simon, Fix Jérémy, Pradalier Cédric, Mathieu Laurence
Université de Lorraine, CNRS, LCPME, F-54000, Nancy, France.
Université de Lorraine, CNRS, LEMTA, F-54000, Nancy, France.
Biofilm. 2024 Nov 16;8:100240. doi: 10.1016/j.bioflm.2024.100240. eCollection 2024 Dec.
Understanding pioneer bacterial adhesion is essential to appreciate bacterial colonization and consider appropriate control strategies. This bacterial entrapment at the wall is known to be controlled by many physical, chemical or biological factors, including hydrodynamic conditions. However, due to the nature of early bacterial adhesion, i.e. a short and dynamic process with low biomass involved, such investigations are challenging. In this context, our study aimed to evaluate the effect of wall shear rate on the early bacterial adhesion dynamics. Firstly, at the population scale by assessing bacterial colonization kinetics and the mechanisms responsible for wall transfer under shear rates using a time-lapse approach. Secondly, at the individual scale, by implementing an automated image processing method based on deep learning to track each individual pioneer bacterium on the wall. Bacterial adhesion experiments are performed on a model bacterium ( MR-1) at different shear rates (0 to1250 s) in a microfluidic system mounted under a microscope equipped with a CCD camera. Image processing was performed using a trained neural network (YOLOv8), which allowed information extraction, i.e. bacterial wall residence time and orientation for each adhered bacterium during pioneer colonization (14 min). Collected from over 20,000 bacteria, our results showed that adhered bacteria had a very short residence time at the wall, with over 70 % remaining less than 1 min. Shear rates had a non-proportional effect on pioneer colonization with a bell-shape profile suggesting that intermediate shear rates improved both bacterial wall residence time as well as colonization rate and level. This lack of proportionality highlights the dual effect of wall shear rate on early bacterial colonization; initially increasing it improves bacterial colonization up to a threshold, beyond which it leads to higher bacterial wall detachment. The present study provides quantitative data on the individual dynamics of just adhered bacteria within a population when exposed to different rates of wall shear.
了解先锋细菌的粘附对于认识细菌定殖并考虑适当的控制策略至关重要。已知这种细菌在壁上的截留受许多物理、化学或生物因素控制,包括流体动力学条件。然而,由于早期细菌粘附的性质,即一个涉及生物量低的短暂且动态的过程,此类研究具有挑战性。在此背景下,我们的研究旨在评估壁面剪切速率对早期细菌粘附动力学的影响。首先,在群体尺度上,通过使用延时方法评估细菌定殖动力学以及在剪切速率下负责壁面转移的机制。其次,在个体尺度上,通过实施基于深度学习的自动图像处理方法来跟踪壁面上的每一个先锋细菌个体。在配备电荷耦合器件(CCD)相机的显微镜下安装的微流控系统中,对模型细菌(MR-1)在不同剪切速率(0至1250秒⁻¹)下进行细菌粘附实验。使用经过训练的神经网络(YOLOv8)进行图像处理,该网络允许提取信息,即先锋定殖(14分钟)期间每个粘附细菌的壁面停留时间和取向。从20000多个细菌中收集的数据表明,粘附细菌在壁面上的停留时间非常短,超过70%的细菌停留时间不到1分钟。剪切速率对先锋定殖具有非比例效应,呈钟形曲线,这表明中等剪切速率既能提高细菌在壁面上的停留时间,又能提高定殖速率和水平。这种非比例性突出了壁面剪切速率对早期细菌定殖的双重作用;最初增加剪切速率可提高细菌定殖,直至达到一个阈值,超过该阈值则会导致更高的细菌从壁面脱落。本研究提供了关于群体中刚粘附细菌在暴露于不同壁面剪切速率时个体动态的定量数据。