Innovative methodology for antimicrobial susceptibility determination in Mycoplasma biofilms

Mycoplasma spp. are facultative pathogens that contribute to the pathogenesis of multiple bovine diseases including the bovine respiratory disease complex and that have been shown to form biofilms. Biofilm formation is associated with increased antibiotic resistance in many organisms, but accurate determination of antimicrobial susceptibility in biofilms is challenging. In Mycoplasma spp., antimicrobial susceptibility is routinely determined using metabolic pH-dependent color change. However, biofilm formation generally leads to reduced metabolism, making interpretation of metabolic readouts difficult. Therefore, we developed and optimized a new flow-cytometry-based method for antimicrobial susceptibility testing in biofilm-forming Mycoplasma, termed the live-dead antimicrobial susceptibility test (LD-AST). The LD-AST measures the proportion of dead bacteria upon exposure to antibiotics, works robustly with both planktonic and biofilm cultures, and enables the determination of the minim..., , , # Data from: Innovative methodology for antimicrobial susceptibility determination in M. bovis biofilms

[https://doi.org/10.5061/dryad.m905qfv8m](https://doi.org/10.5061/dryad.m905qfv8m)

## Description of the data and file structure

### 2\_4\_Confluence\_and\_Structure\_Size.csv was generated using the methods described in section 2.4

*Isolate*: *Mycoplasma bovis* or *Mycoplasma bovoculi* isolate name.
*Percent Area*: The confluence of the biofilm on the field of view expressed as percent area.
*Day*: The time point post inoculation expressed in days.
*Well*: The well of the 96-well plate that the reading corresponds to.
*plate.well*: The concatenated individual IDs for both the well within individual plates.
*measurement*: The measurement in micrometers of the largest observed structure.

The data points where there are NAs for measurement are due to the absence of large structures observed at the time points.

### 2\_5\_pH\_Experiment\_Compiled\_Data.csv was generated using the me...