Abstract Cardiovascular disease in dogs may have minimal outward signs in the beginning stages, but early detection and intervention are vital to help affected dogs live longer and have better quality of life. N-terminal pro-B-type natriuretic peptide (NT-proBNP) and troponin-I are two biomarkers that show promise for detection of cardiovascular disease at a research level, but a deeper understanding is necessary to bring them into the forefront for diagnostic purposes. The aim of this research was to present values of NT-proBNP and troponin-I in healthy Labrador retrievers and in Labrador retrievers with subclinical dilated cardiomyopathy (subDCM). 55 intact Labrador retrievers (27m/28f) were selected based on good clinical health, with no prior evidence of cardiovascular disease. All dogs were maintained on the same standard kennel diet prior to testing. Each dog had plasma collected for NT-proBNP analysis (Cardiopet; Idexx) and serum collected for troponin-I analysis (Immulite; UC Davis Vet Lab). Each dog then had an echocardiogram performed by an experienced sonographer which was reviewed by a boarded veterinary cardiologist. Out of 55 adult Labrador retrievers, 46 (24m/22f) were classified as normal and 9 (3m/6f) were classified as having occult subDCM. Normal Labrador retrievers had a significantly lower mean NT-proBNP of 695 ± 432 pmol/L compared to subDCM Labrador retrievers with a mean of 1144 ± 546 pmol/L (P = 0.009). Normal Labrador retrievers had a lower mean troponin-I of 0.07 ± 0.04ng/mL compared to subDCM Labrador retrievers with a mean of 0.1 ± 0.07ng/mL, trending towards significance (P = 0.08). In summary, healthy Labrador retrievers had significantly lower NT-proBNP compared to Labrador retrievers affected with subclinical dilated cardiomyopathy.