Experiments were conducted to determine the effects of Newcastle disease on chicken fat metabolism. Thirty black-bone chickens were infected intraocularly with the Newcastle disease virus (NDV). Six birds were killed at 0, 12, 24, 48 and 72 h post infection, respectively. Results showed that the NDV infection decreased concentration of high-density lipoprotein cholesterol and increased concentrations of total cholesterol and low-density lipoprotein cholesterol in the plasma. Concentrations of triglycerides and free fatty acid were decreased after their initial increase. NDV infection also dramatically raised the activities of lipoprotein lipase (LPL), hepatic lipase and lipases in the serum. Furthermore, PCR results showed that the incipient infection up-regulated mRNA expression of LPL, adipose triglyceride lipase and nuclear factor peroxisome proliferator-activated receptor alpha (PPARα), but down-regulated them at later stage. Similarly, mRNA expression of fatty acid synthase, acetyl-CoA carboxylase and nuclear factor PPARγ, fatty acid transport protein 1 (FATP1), and 4(FATP4) decreased, whereas fatty acid translocase and fatty acid-binding protein increased initially. Data from Western blotting analysis showed that the changes in protein levels were consistent with mRNA expression. These results indicated that fat metabolism of the chicken was affected by the NDV infection. At the beginning of NDV infection, lipogenesis was inhibited, whereas lipolysis was strengthened. After lipolysis was strengthened, fat metabolism was found to be maximally depressed.