Coresistance to human reserve antibiotics can be selected by antibiotics used in veterinary medicine. A Clostridium perfringens strain isolated from pig manure was resistant to the reserve drug linezolid and, simultaneously, resistant against florfenicol and erythromycin. We detected a new mutation in a highly conserved region of rplD, encoding protein L4 of the 50S ribosomal subunit. This is the first genetic substantiation of linezolid resistance in the genus Clostridium.