Inappropriate antibiotic use is a major factor contributing to the emergence and spread of antimicrobial resistance. The long turnaround time (over 24 hours) required for clinical antimicrobial susceptibility testing (AST) often results in patients being prescribed empiric therapies, which may be inadequate, inappropriate, or overly broad-spectrum. A reduction in the AST time may enable more appropriate therapies to be prescribed earlier. Here we report on a new diagnostic asynchronous magnetic bead rotation (AMBR) biosensor droplet microfluidic platform that enables single cell and small cell population growth measurements for applications aimed at rapid AST. We demonstrate the ability to rapidly measure bacterial growth, susceptibility, and the minimum inhibitory concentration (MIC) of a small uropathogenic Escherichia coli population that was confined in microfluidic droplets and exposed to concentrations above and below the MIC of gentamicin. Growth was observed below the MIC, and no growth was observed above the MIC. A 52% change in the sensor signal (i.e. rotational period) was observed within 15 minutes, thus allowing AST measurements to be performed potentially within minutes.
Continuous growth of individual bacteria has been previously studied by direct observation using optical imaging. However, optical microscopy studies are inherently diffraction limited and limited in the number of individual cells that can be continuously monitored. Here we report on the use of the asynchronous magnetic bead rotation (AMBR) sensor, which is not diffraction limited. The AMBR sensor allows for the measurement of nanoscale growth dynamics of individual bacterial cells, over multiple generations. This torque-based magnetic bead sensor monitors variations in drag caused by the attachment and growth of a single bacterial cell. In this manner, we observed the growth and division of individual E. coli bacteria, with 80 nanometer sensitivity to the cell length. Over the life cycle of a cell we observed up to 300 % increase in the rotational period of the biosensor due to increased cell volume. In addition, we observed single bacterial cell growth response to antibiotics. This work demonstrates a non-microscopy based approach for monitoring individual cell growth dynamics, including cell elongation, generation time, lag time, and division, as well as their sensitivity to antibiotics. † Corresponding authors: Raoul Kopelman kopelman@umich.edu and Brandon McNaughton bmcnaugh@umich.edu,
Mine tailings account for most of the environmental incidents related to the extractive industry, with risks increasing due to steadily rising tonnage of low-grade ore and extreme weather events. Recycling of tailings in raw-material-intensive applications presents an interesting alternative to costly tailings management with associated restoration efforts. Chemically bonded ceramics may offer a route to upgrading mine tailings into raw materials for ceramics. In this review such chemically bonded ceramic methods that may be used to recycle mine tailings as raw materials, are reviewed while focusing in particular on two methods: 1) geopolymerization/alkali activation and 2) chemically bonded phosphate ceramics. The aim of the review is not to give exhaustive review on the wide topic, but to scope the required boundary conditions that need to be met for such utilization. According to the findings, alkali activation has been studied for 28 separate silicate minerals in the scientific literature, and presents a viable method, which is already in commercial use in calcium-rich cement-like binder applications. Phosphate bonding literature is more focused on phosphate containing minerals and waste encapsulation. Very little work has been done on low-calcium tailings utilization with either technology, and more knowledge is needed on the effect of different pretreatment methods to increase reactivity of mine tailings in chemically bonded ceramics.
The long turnaround time in antimicrobial susceptibility testing (AST) endangers patients and encourages the administration of wide spectrum antibiotics, thus resulting in alarming increases of multi-drug resistant pathogens. A method for faster detection of bacterial proliferation presents one avenue towards addressing this global concern. We report on a label-free asynchronous magnetic bead rotation (AMBR) based viscometry method that rapidly detects bacterial growth and determines drug sensitivity by measuring changes in the suspension’s viscosity. With this platform, we observed the growth of a uropathogenic Escherichia coli isolate, with an initial concentration of 50 cells per drop, within 20 minutes; in addition, we determined the gentamicin minimum inhibitory concentration (MIC) of the E. coli isolate within 100 minutes. We thus demonstrated a label-free, micro-viscometer platform that can measure bacterial growth and drug susceptibility more rapidly, with lower initial bacterial counts than existing commercial systems, and potentially with any microbial strains.
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