The anaerobic, spore-forming pathogen Clostridioides difficile infects the gastrointestinal tract of higher mammals, including humans, and is the major causative agent of antibiotic-associated diarrhea. Moreover, recurrence and re-infection rates after a first successful antibiotic therapy are substantially high. To overcome the problem of recurrence and re-infection new antibiotics are urgently required that are more selective for the pathogen but spare other members of the microbiota. In turn, the microbial community inside the intestine is allowed to recover and to re-establish colonization resistance to protect against recurrence. The data presented here aim at analyzing the ability of the macrolide compound chlorotonil A to meet the requirements of C. difficile specific drug.