Laying hens' metabolism goes through a cyclic process to produce eggs, which requires higher dietary protein and energy in the morning (AM) and higher calcium (Ca) in the afternoon/evening (PM) than the rest of the day. Therefore, poultry scientists are trying to adopt a new feeding strategy called AM/PM or split feeding to precisely meet hen's requirements more effectively than conventional methods. A 10-week cage layer trial was carried out via a Box-Behnken response surface design to identify the optimal amount of protein, energy, and calcium of the AM/PM diets. There were 13 test treatments with three levels of crude protein (19.6%/18.4%, 20.3%/17.7%, 21%/17%), calcium (3.3%/4.9%, 2.5%/5.7%, 1.6%/6.6%), and apparent metabolizable energy (AME) (12 MJ/kg/11.2 MJ/kg, 12.4 MJ/kg/10.8 MJ/kg, 12.8 MJ/kg/10.4 MJ/kg) for AM/PM diets respectively and a control treatment with industry baseline (CP-19%, Ca-4.1% and ME 11.6 MJ/kg). These are the calculated values of nutrients on a dry matter basis. A total of 364 hens were randomly distributed into 2 dietary treatments where each treatment had 13 replicates (2 hens per replicate cage, 26 hens per treatment). AM and PM diets were swapped out at approximately 8 am and 4 pm each day. Egg production and hen performance were measured daily and weekly, respectively, with egg quality, serum Ca, and nutrient digestibility measured at week 10. AM:PM intake and feed cost were calculated for each treatment. The optimal FCR, feed cost, and AM:PM intake were used to determine Ca, CP, and AME levels. The result showed that 6 out of 13 of our test treatments gave improved FCR compared to the control treatment (p = 0.017). Dietary treatments did not affect overall hen weight and serum Ca and egg quality at week 10, except for the lower yolk color score in the control treatment (p = 0.002). Hens in the experimental treatment, with calcium levels of 1.6% and 6.6%, crude protein (CP) levels of 19.6% and 18.4%, and AME content of 12.4 MJ/kg and 10.8 MJ/kg in the AM and PM diet, respectively, showed the highest apparent protein digestibility (56.6%) compared to the control group (p < 0.05). Similarly, hens receiving a treatment containing calcium at 3.3% and 4.9%, CP at 21% and 17%, and AME at 12.4 MJ/kg and 10.8 MJ/kg in the AM and PM diet, respectively, achieved the highest calcium digestibility (62.13%), while the control treatment yielded the lowest calcium digestibility (p < 0.05). After analyzing the data using the Box-Behnken response surface methodology, we found that (21/17)% CP, (3.3/4.9)% Ca, and (12/11.12) MJ/kg energy in the AM/PM diet gave the optimum performance in terms of lower feed cost and better feed efficiency. The data of AM:PM intake demonstrate that selective feeding occurs in between treatments (p < 0.001) and the degree of selection depends on the difference between the level of nutrients in AM and PM diet. This study revealed that when optimized, AM/PM feeding improves feed efficiency and egg quality of laying hens.
