Silver Ions Inhibit Bacterial Movement and Stall Flagellar Motor.

Silver (Ag) in different forms has been gaining broad attention due to its antimicrobial activities and the increasing resistance of bacteria to commonly prescribed antibiotics. However, various aspects of the antimicrobial mechanism of Ag have not been understood, including how Ag affects bacterial motility, a factor intimately related to bacterial virulence. Here, we report our study on how Ag+ ions affect the motility of E. coli bacteria using swimming, tethering, and rotation assays. We observed that the bacteria slowed down dramatically by >70% when subjected to Ag+ ions, providing direct evidence that Ag+ ions inhibit the motility of bacteria. In addition, through tethering and rotation assays, we monitored the rotation of flagellar motors and observed that the tumbling/pausing frequency of bacteria increased significantly by 77% in the presence of Ag+ ions. Furthermore, we analyzed the results from the tethering assay using the hidden Markov model (HMM) and found that Ag+ ions decreased bacterial tumbling/pausing-to-running transition rate significantly by 75%. The results suggest that the rotation of bacterial flagellar motors was stalled by Ag+ ions. This work provided a new quantitative understanding of the mechanism of Ag-based antimicrobial agents in bacterial motility.

Bacterial mobility and motility in porous media mimicked by microspheres.

Bacterial motion in porous media is essential for their survival, proper functioning, and various applications. Here we investigated the motion of Escherichia coli bacteria in microsphere-mimicked porous media. We observed reduced bacterial velocity and enhanced directional changes of bacteria as the density of microspheres increased, while such changes happened mostly around the microspheres and due to the collisions with the microspheres. More importantly, we established and quantified the correlation between the bacterial trapping in porous media and the geometric confinement imposed by the microspheres. In addition, numerical simulations showed that the active Brownian motion model in the presence of microspheres resulted in bacterial motion that are consistent with the experimental observations. Our study suggested that it is important to distinguish the ability of bacteria to move easily - bacterial mobility - from the ability of bacteria to move independently - bacteria motility. Our results showed that bacterial motility remains similar in porous media, but bacterial mobility was significantly affected by the pore-scale confinement.

Bacterial Growth Curve Measurements with a Multimode Microplate Reader.

Bacterial studies based on growth curves are common in microbiology and related fields. Compared to the standard photometer and cuvette based protocols, bacterial growth curve measurements with microplate readers provide better temporal resolution, higher efficiency, and are less laborious, while analysis and interpretation of the microplate-based measurements are less straightforward. Recently, we developed a new analysis method for evaluating bacterial growth with microplate readers based on time derivatives. Here, we describe a detailed protocol for this development and provide the homemade program for the new analysis method.

Interactions of E. coli with cylindrical micro-pillars of different geometric modifications.

Understanding the behavior of bacteria at the proximity of different surfaces is of great importance and interest. Despite recent exciting progress in geometric control of bacterial behavior around surfaces, a detailed comparison on the interaction of bacteria with cylindrical surfaces of different geometric modifications is still missing. Here, we investigated how bacteria interacted with cylindrical micro-pillars and modified cylindrical micro-pillars with sprocket, gear, and flower-like wall surface features. Using phase-contrast microscopy, we examined the motion of bacteria around the micro-pillars, and observed different responses of bacteria to each geometric modification. In addition, we extracted the trajectories of the bacteria and characterized several parameters (instantaneous velocity v, change of direction δ, approaching angle ϕ) to quantitatively compare the effects of the geometric modifications on the micro-pillars. We found that sharp spikes showed the largest effect, compared to smooth surface, convex and concave ripples. Lastly, we carried out numerical simulations, which explained the experimental observations and showed that the observed effects were due to the geometric modifications.

Microampere Electric Current Causes Bacterial Membrane Damage and Two-Way Leakage in a Short Period of Time.

Physical agents, such as low electric voltage and current, have recently gained attention for antimicrobial treatment due to their bactericidal capability. Although microampere electric current was shown to suppress the growth of bacteria, it remains unclear to what extent the microampere current damaged the bacterial membrane. Here, we investigated the membrane damage and two-way leakage caused by microampere electric current (≤100 µA) with a short exposure time (30 min). Based on MitoTracker staining, propidium iodide staining, filtration assays, and quantitative single-molecule localization microscopy, we observed significant membrane damage, which allowed two-way leakage of ions, small molecules, and proteins. This study paves the way to new development of antimicrobial applications for ultralow electric voltage and current.IMPORTANCE Although electric voltage and current have been studied for a long time in terms of their ability to suppress the growth of bacteria and to kill bacteria, increasing interest has been aroused more recently due to the prevalence of antibiotic resistance of microbes in past decades. Toward understanding the antimicrobial mechanism of low electric voltage and current, previous studies showed that treating bacteria with milliampere electric currents (≥5 mA) for ≥72 h led to significant damage of the bacterial membrane, which likely resulted in leakage of cellular contents and influx of toxic substances through the damaged membrane. However, it remains unclear to what extent membrane damage and two-way (i.e., inward and outward) leakage are caused by lower (i.e., microampere) electric current in a shorter time frame. In this work, we set out to answer this question. We observed that the membrane damage was caused by microampere electric current in half an hour, which allowed two-way leakage of ions, small molecules, and proteins.