A total of 196 day-old male broiler chicks were randomly allocated to 1 of 4 treatments in a study conducted to determine the effects of dietary supplementation of chito-oligosaccharide (COS) on growth, nutrient digestibility, and serum composition. The experimental diets consisted of an unsupplemented control diet based on corn, soybean meal, and fish meal or similar diets supplemented with either chlortetracycline, 50 mg/kg of COS, or 100 mg/kg of COS. Each treatment was fed to 7 replicate pens of birds, with 7 birds per pen. Broiler performance, nutrient digestibility, cecal microbial concentrations, and serum indices were measured at the end of the starter (d 21) and grower phases (d 42). During the starter period and overall, broilers fed 50 or 100 mg/kg of COS had better (P<0.05) average daily gain, average daily feed intake, and feed conversion than the control birds. The performance of birds fed chlortetracycline was generally intermediate between that of the control and the 2 COS treatments. Compared with the birds in the control or chlortetracycline treatments, the birds receiving 100 mg/kg of COS had better nutrient digestibility of DM, energy, calcium, and phosphorus; higher (P<0.05) concentrations of cecal Lactobacillus; and lower (P<0.05) serum triglyceride and total cholesterol during the starter phase. During the grower phase, the birds fed 100 mg/kg of COS had higher (P<0.05) calcium digestibility and CP retention than those fed the chlortetracycline treatment, and lower concentrations of cecal Escherichia coli than birds in the control treatment. The serum growth hormone level in birds fed 50 mg/kg of COS was higher (P<0.05) than in the other treatments. The birds fed 100 mg/kg of COS had lower (P<0.05) serum triglyceride, higher (P<0.05) serum high-density lipoprotein cholesterol, and higher serum total protein content than birds in the other treatments. In conclusion, dietary supplementation with COS appeared to improve the average daily gain of broilers by increasing the average daily feed intake and nutrient digestibility and modulating the concentrations of cecal microbial flora. Additionally, COS increased serum protein and high-density lipoprotein cholesterol and decreased serum triglyceride.