Z-Rings as a Biomarker in Persister Resuscitation

Bacterial infections that survive anti-microbial treatments are a growing concern in the medical field. Much of this research is focused on antibiotic resistance, but bacterial persistence is an equal contributor to the problem. Persister cells are a subpopulation of bacteria that are transiently capable of surviving antibiotics. The surviving bacteria remain after treatment and can resuscitate causing chronic infections. To research resuscitation in these persistent cells properly, we first need to be able to identify which cells to study; using biomarkers is one way to determine which persister cells are capable of resuscitation. One possible biomarker is the formation of z-rings during the growth-inhibited state. Using both Gentamicin and Streptomycin antibiotics, the length of time needed to kill off all but persistent cells is determined using a biphasic kill curve generated using survival assays. The length of treatment is used to treat a culture until only persisters remain. The persisters are then starved in PBS for two days to allow time for the formation Z-rings. The bacteria are then moved onto a microscope slide lined with LB agar which provides nutrients to facilitate resuscitation. Pictures in both phase contrast and gfp fluorescence are taken every hour after plating to monitor the growth and Z-rings. If the formation of Z-rings in the growth-inhibited state holds true as a biomarker, then we can use them to determine which cells are viable for resuscitation study; these cells can then be used to advance the field of persistence research further.