Objective: To assess B-mode and M-mode echocardiographic reference values in Domestic rabbits (Oryctolagus cuniculus). Animals: 100 healthy domestic rabbits. Procedures: M-mode and B-mode measurements of the LVFWd, AO, LA, LA/AO, LVFWs, LVIDd, LVIDs, IVSd, IVSs, and HR were determined. Results: Mean±SD heart rate during echocardiographic tests was 204.33±3.929 beats/min. Mean±SD measurements in diastole and systole for the interventricular septum thickness, left ventricular internal diameter, and left ventricular free wall thickness were 2.5496±0.125 mm and 3.4232±0.134 mm; 15.5428±0.579 mm and 10.9706±0.439 mm; and 2.355±0.0977 mm and 3.6119±0.155 mm, respectively. Mean ± SD left atrial-to-aortic diameter ratio was 1.2265 ±0.0202, and Mean ± SD for fractional shortening and ejection fraction were 29.6047±0.941% and 60.0756±1.328%, respectively. All echocardiographic parameters were studied separately in males and females and in different weights. Conclusions and Clinical Relevance: Echocardiographic reference values for non-anesthetized domestic rabbits are presented providing reference values for future studies. Most of the results were comparable to those reported in non-anesthetized rabbits and may offer a good alternative when this data is necessary.