A high-throughput anaerobic method for viability assays

The routine assessment for the viability of anaerobes is based on bacterial plating, but so far, it has been limited in throughput by the long preparation steps and the tedious anaerobic culturing. Thus, comparatively little is known about the susceptibility pattern, and the sporulation of anaerobes because of the absence of the proper method. Here, we show GVA can quantify the anaerobic <i>Clostridiums</i> colonies accurately by utilizing an anaerobic jar to measure viable cells and spores in high throughput with minimal volumes of reagents and at a comparable time to the traditional viability testing practice. Furthermore, this method enabled high-throughput detection of the bactericidal activity of the antibiotics against anaerobes and allowed for the quantification of hetero-tolerant/resistant subpopulation, which was previously unattainable. Our approach is rapid and easy to use, making it ideal for various applications where high-throughput capabilities can drive innovation, including drug-microbe interactions, host-microbe interactions, and microbe-microbe interactions.