SummaryThis experiment was conducted to evaluate the effects of nanocalcium carbonate (NCC) instead of calcium carbonate (CC) on egg production, egg weight, egg mass, FCR, blood calcium and egg quality characteristics in laying hens. A total of 120 laying hens were used in a 10-weeks trial, from week 23 to 33 of age. Laying hens were randomly assigned to six treatments with four replications, five hens each. The experimental treatments involved replacing 50% of the CC in the diet by decreasing amounts of NCC and were T1 Basal diet (BD) with 8.06% CC; T2 (6.045% of CC as a negative control); T3 (4.03% of CC replaced by 2.015% NCC); T4 (4.03% of CC replaced by 1.01% NCC); T5 (4.03% of CC replaced by 0.252% NCC) and T6 (4.03 of CC replaced with 0.126%NCC).Egg weight was unaffected by dietary treatments (p > .05).However, the egg production percentage and egg mass in T6 were less than that of other treatments (p < .05). The laying hens in the control group had the best average feed conversion ratio (p < .05). Also, the lowest concentration of calcium in hens' blood was recorded for birds fed T6 (p < .05). The best egg shell quality (relative egg shell weight and egg shell weight/surface) was observed in T1 (p < .05).Collectively, our results demonstrated that NCC could replace CC at a lower inclusion level but extreme reduction of calcium concentration in diets (to 1.43% Ca in the T6 group) reduced production performance, egg quality characteristics, Tibia thickness and blood calcium of laying hens. K E Y W O R D Slaying hen, nanocalcium carbonate (NCC), performance, plasma calcium