Background and Aim: The exchange of oxygen and carbon dioxide (CO2) in the incubator plays a key role in embryonic development and hatching. This study aimed to study the effect of non-ventilated (NV) intervals during the early stage of embryonic development on the hatching process. Materials and Methods: Hatching eggs (n = 7200) were equally divided into four treatment groups and incubated in four incubators. The first group was incubated in normal ventilated condition (V) during the setting phase of incubation. Ventilation holes of the three remaining incubators were closed for the first 3, 6, and 9 days and termed as NV groups (NV1, NV2, and NV3, respectively). A gradual increase in CO2 was allowed for NV groups, followed by opening the incubator holes to permit ventilation throughout the rest of the incubation periods. Results: Obtained results demonstrated that CO2 concentration gradually increased up to 0.19% for the NV1 group, 0.41% for the NV2 group, and 0.90% for the NV3 group, while CO2 concentration remained at 0.08% during the first 9 days of incubation in the V group. Albumen pH was lowered for all NV groups. The highest hatchability percentage was recorded for NV3 followed by NV2 and NV1 groups. All NV groups represented earlier and narrower spread of hatch and higher hatched chick weight. Embryos and hatched chicks in the NV groups had higher hormonal levels of thyroxin and corticosterone. Conclusion: All non-ventilation periods had positive effects on narrowing the spread of hatch, increasing hatched chick weight and hatchability percentage compared to the normal V condition. Furthermore, the non-ventilation throughout the first 9 days of incubation yielded the best hatching results. Keywords: albumen pH, carbon dioxide, embryonic hormones, hatchability, non-ventilation.

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