The effect of probiotic on egg production, egg quality, yolk cholesterol, triglyceride and blood biochemical parameters of layer hens was studied. Experiment was carried out with 256 bird and each sixteen bird were considered as a replicate that were randomly allotted to basal with Probiotic supplementation at rates of 25, 50 and 75 mg/ kg diets. Treatments were arranged in a completely randomized design. The Probiotic used in the study was a commercial preparation (Protexin) with 3× 109 spore g-1 of the product. Egg production, egg weight, feed intake, feed conversion ratio and egg mass were not influenced by Probiotic, as well as Haug unit, specific gravity, egg shell weight, egg shell thickness and shell weight/ surface area. Experimental groups had not statistical differences in biochemical parameters such as albumin, total protein, phosphorus, calcium, cholesterol, triglyceride and glucose amounts. Probiotic had not also any effect on egg yolk cholesterol and triglyceride. It was concluded that dietary Probiotic supplementation up to 75 mg/kg of diet had not beneficial effects on the performance and egg quality of laying hens.