Rolling the evolutionary dice:Neisseriacommensals as proxies for elucidating the underpinnings of antibiotic resistance mechanisms and evolution in human pathogens

Abstract

ABSTRACTSpecies within the genusNeisseriaare especially adept at sharing adaptive allelic variation across species’ boundaries, with commensal species repeatedly transferring resistance to their pathogenic relativeN. gonorrhoeae. However, resistance in commensalNeisseriais infrequently characterized at both the phenotypic and genotypic levels, limiting our ability to predict novel and potentially transferable resistance mechanisms that ultimately may become important clinically. Unique evolutionary starting places of eachNeisseriaspecies will have distinct genomic backgrounds, which may ultimately control the fate of evolving populations in response to selection, as epistatic and additive interactions may coerce lineages along divergent evolutionary trajectories. However alternatively, similar genetic content present across species due to shared ancestry may constrain the adaptive solutions that exist. Thus, identifying the paths to resistance across commensals may aid in characterizing theNeisseriaresistome – or the reservoir of alleles within the genus, as well as its depth. Here, we usein vitroevolution of four commensal species to investigate the potential for and repeatability of resistance evolution to two antimicrobials, the macrolide azithromycin and the β-lactam penicillin. After 20 days of selection, commensals evolved elevated minimum inhibitory concentrations (MICs) to penicillin and azithromycin in 11/16 and 12/16 cases respectively. Almost all cases of resistance emergence converged on mutations within ribosomal components or themtrRCDEefflux pump for azithromycin-based selection, andmtrRCDEorpenAfor penicillin selection; thus, supporting constrained adaptive solutions despite divergent evolutionary starting points across the genus for these particular drugs. However, continuing to explore the paths to resistance across different experimental conditions and genomic backgrounds, which could shunt evolution down alternative evolutionary trajectories, will ultimately flesh out the fullNeisseriaresistome.