Changes in bacterial physiology necessarily precede cell death in response to antibiotics.Herein we investigate the early silver lining herbs kidney support disruption of Ca2+ homeostasis as a marker for antibiotic response.Using a machine learning framework, we quantify the temporal information encoded in single-cell Ca2+ dynamics.We find Ca2+ dynamics distinguish kanamycin sensitive and resistant cells before changes in gross cell phenotypes such as cell growth or protein stability.The onset time (pharmacokinetics) and probability (pharmacodynamics) of these aberrant Ca2+ dynamics are dose and time-dependent, even at the resolution of single-cells.
Of the compounds profiled, we find Ca2+ dynamics are also an indicator of Polymyxin B activity.In Polymyxin B treated cells, we find aberrant Ca2+ dynamics precedes the entry of propidium iodide marking membrane permeabilization.Additionally, we find modifying membrane voltage and external Ca2+ concentration alters the time between these aberrant dynamics and membrane breakdown suggesting a previously unappreciated role of Ca2+ in the membrane destabilization during Polymyxin B treatment.In conclusion, leveraging live, single-cell, Ca2+ imaging coupled with machine learning, we have demonstrated the discriminative capacity of Ca2+ dynamics in osborne hog feeders for sale identifying antibiotic-resistant bacteria.