The Effect of the Stringent Response and Oxidative Stress Response on Fitness Costs of De Novo Acquisition of Antibiotic Resistance  
[PERSON]  
1 Laboratory for Molecular Biology and Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands; [EMAIL_ADDRESS] (W.Q. ); [EMAIL_ADDRESS] (D.K. ); [EMAIL_ADDRESS] (M.W.) [PERSON]  
2 RNA Biology & Applied Bioinformatics, Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands; [EMAIL_ADDRESS] (M.J.J. ); [EMAIL_ADDRESS] (W.d.L.) [PERSON]  
1 Laboratory for Molecular Biology and Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands; [EMAIL_ADDRESS] (W.Q. ); [EMAIL_ADDRESS] (D.K. ); [EMAIL_ADDRESS] (M.W.) [PERSON]  
2 RNA Biology & Applied Bioinformatics, Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands; [EMAIL_ADDRESS] (M.J.J. ); [EMAIL_ADDRESS] (W.d.L.) [PERSON]  
1 Laboratory for Molecular Biology and Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands; [EMAIL_ADDRESS] (W.Q. ); [EMAIL_ADDRESS] (D.K. ); [EMAIL_ADDRESS] (M.W.) [PERSON]  
1 Laboratory for Molecular Biology and Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH Amsterdam, The Netherlands; [EMAIL_ADDRESS] (W.Q. ); [EMAIL_ADDRESS] (D.K. ); [EMAIL_ADDRESS] (M.W.) Footnote 21: email: {w.  
bacterial fitness [6; 7; 8]. Conversion of cellular homeostasis systems, such as ion- and pH-pumps, into antibiotic efflux pumps causes a reduced capability to control the intracellular environment [9]. Mutations within genes associated with resistance, such as the modification of drug targets, degradation of antibiotics, reduced antibiotic uptake, and increased efflux, can result in diminished survival and replicative capabilities among resistant populations [4]. Consequently, it has been suggested that limiting antibiotic usage, and, thus, reducing the selection of pressure-favoring resistant populations, may allow more adaptable and susceptible populations to outcompete and, ultimately, eradicate resistance [10]. However, several other responses of bacteria becoming resistant, such as the co-selection of resistance genes alongside other functional traits, can provide an adaptive advantage [11]. The acquisition of resistance plasmids did not reduce growth rates in _E. coli_[12]. Low-cost resistance mutations and compensatory evolution can compensate for the initial metabolic costs [10]. Therefore, a comprehensive understanding of the intricate relationship between mutation occurrence and fitness costs is essential for addressing the persistent challenge of antibiotic resistance. Under antibiotic stress, bacteria initiate a variety of stress responses such as stringent and oxidative stress responses [13]. Bacterial stringent and oxidative stress responses are involved in the development of antibiotic resistance [14; 15]. Bactericidal antibiotics interact with their targets in a way that increases the oxidation of NADH and thus produces respiratory chain byproducts such as reactive oxygen species (ROS) [16]. Although ROS at high concentrations kills bacteria, DNA damage caused by sub-lethal levels of ROS can activate the repair system induced by the SOS response, thereby increasing the mutation rate [14; 17]. Ultimately, exposure to antibiotics can cause mutations in antimicrobial resistance-related genes. The single-gene knockout strain of (p)ppGpp synthase ReIA has a reduced stringent response, resulting in lower growth rates and reduced ROS formation, reducing the development of drug resistance [15]. As these two stress responses also affect the growth rate, it is unclear to what extent the fitness costs during the development of resistance are related to the stress responses.