Betaine functions as an osmoregulators in the cells and its inclusion in diet can spare the choline and carcass fat reduction in chicken. Thus, two hundred eighty eight laying hens were fed with 0, 500, 1,000, 2,000 ppm of betaine from seventy eight to eighty six weeks of age during the environmentally high temperature stress. Com and soybean basal diets contained 2,800 kcal/kg ME and 16% CP. Egg production, feed intake, and feed conversion were examined for eight weeks. Egg quality characters, serum cholesterol, liver betaine, and lower ileal osmolality were measured at the end of experiment. Egg Production rates of hens fed with 500 or 2,000 ppm of betaine were 75.06 and 75.02%, respectively and tended to increase compared to the control. The feed conversion(FCR) of these treatments was significantly(P<0.05) improved compared to that of control although it did not significantly differ in the e99 Production rates between 500 and 2,000 ppm of betain groups. Eggshell breaking strength of hens fed betaine was significantly(P<0.05) higher than those of control. However, betaine supplements did not influence to improving the albumen height and Haugh unit. Liver betaine in hens fed with betaine was linearly increased unto 2,000 ppm. The birds fed with 2,000 ppm betaine showed significantly(P<0.05) higher in the liver betaine than the control birds. Total cholesterol and triglyceride tended to be increased by dietary betaine supplement. The lower ileal osmolality in betaine supplement group tended to be slightly decreased. As a result, dietary betaine supplement tended to improve the egg Production and eggshell Quality in laying hens during heat stress.