The need for appropriate use of antimicrobials limiting the insurgence, selection and spread of bacterial resistance and preventing its diffusion to the community is increasingly widely felt. Based on some CDC expert recommendations, an antibiotic therapy should be driven by the following criteria: scientifically proven clinical evidence, high probability of being beneficial to patients, safety and tolerability, drug and regimen choice made according to the infection to be treated, and cost/efficacy ratio. In this article, we compare the microbiological and pharmacological characteristics of the three most frequently used fluoroquinolones in Italy - ciprofloxacin, levofloxacin, and moxifloxacin - highlighting the main differences and peculiar aspects for each of them. We describe the microbiological profile, providing details on bacterial resistance mechanisms, the pharmacological profile, giving details on tolerability, and the clinical profile, showing the individual therapeutic efficacy, underlining the main differences in their pharmacodynamic and pharmacokinetic aspects. The main national and international guidelines recommend ciprofloxacin and levofloxacin and, to a lesser extent, moxifloxacin, for treating nosocomial infections localised in different organs and apparatuses, including severe infections in critical patients. In addition, they represent a basic therapeutic option for treatment of community-acquired infections, such as urinary and respiratory tract infections.