The effects of dietary supplementation of a commercial probiotic (BioPlus 2B) on daily feed consumption, egg yield, egg weight, specific gravity, body weight, feed conversion ratio, serum and egg yolk cholesterol, and serum trigylceride in layer hens were investigated. In 12 replicates, 480 27-week-old Brown-Nick layers were fed with diets containing 0, 250, 500 or 750 mg kg(-1) probiotic for 90 days. When compared with the controls, supplementation of 250, 500 and 750 mg kg(-1) probiotic increased egg production, but decreased the damaged egg ratio (p < 0.05), egg yolk cholesterol and serum cholesterol (p < 0.001) levels. In addition, serum triglyceride levels were reduced by using 500 and 750 mg kg(-1) probiotic supplementation (p < 0.001). Feed conversion ratios were positively affected by supplementation of 250 and 500 mg kg(-1) probiotic compared with controls (p < 0.05). There was no statistically significant difference between the control and all treatment groups on feed consumption, egg weight, specific gravity, body weight, and egg yolk weight.