BACKGROUND: Homocysteine (Hcy) is a sulfur-containing amino acid derived from methionine. Several studies have suggested that plasma Hcy may be a useful biomarker of cardiovascular disease in human. There is limited information about serum homocysteine in heart diseases of dogs.OBJECTIVES: The present study aimed to assess the concentration of serum total Hcy and evaluate the relationship between its concentrations and serum levels of cholesterol, triglycerides, and activities of creatine phosphokinase and aspartate aminotransferase in small breed of dogs with endocardiosis.METHODS: For this purpose, a total of 15 dogs (seven males and eight females) with acquired endocardiosis were confirmed by clinical examination, auscultation, thoracic radiography, and echocardiography. In addition, 15 normal dogs of the similar age, breed, and weight were selected as the controls. All the examinations performed on the patients were carried out on the healthy dogs. Homocysteine was measured via an enzymatic method. Additionally, the activities of CPK and AST and the levels of cholesterol and triglycerides were assessed utiling commercial kits. Subsequently, the data were analyzed with t- student test and one- way ANOVA and Pearson’s linear correlation test was used to determine the relationship between serum homocysteine levels with triglycerides, cholesterol, CPK, and AST.RESULTS: The results revealed no significant differences concerning serum homocysteine level and AST activity between the two groups; meanwhile, the increase in serum triglycerides and CPK activities in dogs with heart disease was statistically significant compared to that in the control group. However, there was no significant correlation between homocysteine and the other measured parameters.CONCLUSIONS: The present study indicated that the role of homocysteine as a new risk factor associated with dog endocardiosis is questionable and it seems as if in human studies related to homocysteine and heart disease, the selection of dog as animal model requires further investigation.