There is some evidence that Mycoplasma species may be associated with lower airway disease in cats. Retrospective and prospective studies were carried out on a total population of 76 cats but failed to identify any cases of Mycoplasma species infection by bacterial culture alone. The overall prevalence of bacterial infection (15.8%) was also lower than that identified in previous studies. When a molecular detection technique, the PCR-DGGE, was employed the prevalence of Mycoplasma species detected was 15.4%, with M felis, M gateae and M feliminutum species identified, although the significance of these Mycoplasma species in feline lower airway disease remains in question. However, the PCR-DGGE technique allowed species identification and indicated the presence of M feliminutum, a species not previously isolated from the lower airways of cats.