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The objective of the present study was to measure the concentration of Paraoxonase-1 (PON-1) and N-terminal-prohormone-B-type natriuretic peptide (NT-proBNP), in the serum of dogs with degenerative Mitral Valve Disease (MVD), in order to identify their association with the clinical stage and specific clinico-pathologic and echocardiographic findings.Eighty dogs diagnosed with MVD and staged according to the ACVIM (American College of Veterinary Internal Medicine) consensus statement (B1, B2, C and D), based on their clinical, radiographic, and echocardiographic findings, were included in the study. NT-proBNP was measured only in stage B1 and B2 dogs. Clinical stage did not have a significant effect on PON-1 concentrations (<i>p</i> = 0.149), but NT-proBNP levels were lower in B1 dogs (<i>p</i> = 0.001). A significant correlation between PON-1 and total plasma proteins (<i>p</i> = 0.001), albumin (<i>p</i> = 0.003) and white blood cell count (<i>p</i> = 0.041) was detected, whereas there was no significant correlation (<i>p</i> = 0.847) between PON-1 and NT-proBNP concentrations. PON-1 showed a significant but weak negative correlation with normalized left ventricular internal diameter at diastole (LVIDdn) (<i>p</i> = 0.022) and systole (LVIDsn) (<i>p</i> = 0.012), as well as mitral valve E to A wave velocity ratio (MV E/A) (<i>p</i> = 0.015), but not with Left Atrial to Aortic root ratio (LA/Ao) (<i>p</i> = 0.892) or fractional shortening (FS%) (<i>p</i> = 0.944). PON-1 seems to be an insensitive marker of clinical stage and disease severity in MVD, but can be indicative of some clinico-pathological and echocardiographic changes. NT-proBNP changes are independent of oxidative stress.