Upper respiratory infections are a frequent problem in pet rabbits and rodents. Mycoplasma pulmonis is one of the most common causes of respiratory infections in pet rats. There is an older report of detection of M. pulmonis in laboratory rabbits via culture of the nares and oropharynx, but overall Mycoplasma is not routinely considered as a cause of respiratory disease in rabbits. Due to the unexpected detection of Mycoplasma via next-generation sequencing in several clinical cases of rabbits with upper respiratory signs, we set out to investigate the prevalence of Mycoplasma on nasal swabs in pet rabbits. Our hypothesis was that Mycoplasma detection would be relatively common and would be found in both healthy rabbits and rabbits with signs of upper respiratory disease (“unhealthy” rabbits). The results of nasal swabs from both healthy and unhealthy rabbits submitted for next-generation DNA sequencing from January 2022 to February 2023 were reviewed. Data gathered included signalment, whether or not Mycoplasma was detected, and the cell count and relative predominance of Mycoplasma compared to other organisms. Ninety-one rabbits were included, of which 49 were healthy and 42 were unhealthy. Overall, 57.1% (52/91) of rabbits were positive for Mycoplasma, and 42.8% (39/91) were negative for Mycoplasma. Of the healthy rabbits, 75.5% (37/49) were positive for Mycoplasma; of the unhealthy rabbits, 35.7% (15/42) were positive. The fact that Mycoplasma positivity was common in both groups of rabbits, and particularly common in rabbits without upper respiratory signs, suggests that Mycoplasma is normal nasal flora in rabbits.