The aim of this study was to provide two-dimensional (2D) and M-mode echocardiographic reference ranges from a sample of the UK population of donkeys including geriatrics (>30 years), owned by The Donkey Sanctuary, and to assess the influence of gender, weight and age on these variables. A total of 36 donkeys with no clinical or echocardiographic evidence of cardiovascular disease were examined; 24 geldings and 12 females, aged 3–45 years old, weighing 130–262 kg. Left atrial to aortic ratio was larger in geldings (P=0.004). There was no significant difference for left ventricular M-mode diastolic diameter between females and geldings (P=0.121) after exclusion of one heavy female outlier. 2D measurements significantly increased with bodyweight including maximal left atrial diameter (R2=0.112; P=0.046), aortic diameter at various levels (e.g. annulus: R2=0.35; P<0.001) and the pulmonary artery diameter (R2=0.124; P=0.035). M-mode measurements were not significantly influenced by weight other than the left ventricular free wall in systole (R2=0.118; P=0.041). Age and heart rate did not have any significant effect on echocardiographic variables. This is the first UK study to report on echocardiographic reference ranges of sedentary donkeys across a wide age range and shows differences compared with reference ranges from working donkeys.