Abstract This paper reports radial colour and longitudinal mitral annulus pulsed-wave tissue Doppler findings in a large cohort of healthy, adult pet rabbits. Thirty-nine rabbits (22 Dwarf Lops, 14 French Lops and three Alaskans) underwent conscious echocardiography. The median age of the rabbits was 22 months and the median weight was 2.8 kg (Dwarf Lop 2.4 kg/French Lop 6.0 kg). Adequate radial colour and longitudinal pulsed-wave tissue Doppler traces were obtained in 100% and 85% of cases, respectively. Most systolic tissue Doppler parameters were significantly higher in French Lops than in Dwarf Lops. Separation of mitral inflow diastolic waves was present in 40% of cases using conventional spectral Doppler and in >60% of cases using pulsed-wave tissue Doppler which could be beneficial when evaluating diastolic function in rabbits. This study can be used as a reference for normal echocardiographic tissue Doppler values for adult rabbits undergoing conscious echocardiography in clinical practice.