We previously demonstrated that adaptation of an embryo-attenuated infectious bronchitis virus (IBV) Arkansas (Ark) Delmarva Poultry Industry (DPI)–derived vaccine to chicken embryo kidney (CEK) cells (CEKp7) shifted the virus population towards homogeneity in spike (S) and nonstructural protein genes. Moreover, the typical Ark vaccine subpopulations emerging in chickens vaccinated with commercial Ark vaccines were not detected in chickens vaccinated with CEKp7, indicating that kidney-cell adaptation drastically increased the stability of the vaccine virus population in chickens. In the current study both conventional and next-generation sequencing results show that the changes achieved during CEK adaptation remained after five back passages in embryonated chicken egg (ECE). In a first protection study 1-day-old chickens were vaccinated with 10^4.0 or 10^5.0 50% embryo infectious doses (EID₅₀)/chicken of the second ECE back passage of CEKp7 (CEKp7e2) and demonstrated protection against Ark virulent (10^6.0 EID₅₀) challenge. In a second protection trial, protection by CEKp7e2 was compared with protection conferred by an attenuated commercial ArkDPI-derived vaccine different from that which the CEK-adapted virus originated. All vaccinated chicken groups showed a significant reduction of respiratory signs and viral load after Ark virulent challenge compared to unvaccinated-challenged controls. In CEKp7e2 vaccinated chickens viral subpopulations different from the challenge virus were detected after challenge in a marginal number (7%–8%) of chickens. In contrast, IBV S1 sequences that differed from the predominant population in the challenge virus were detected after challenge in a large number (77%) of chickens vaccinated with the commercial Ark attenuated vaccine. The CEK-adapted IBV ArkDPI-derived vaccine is a stable and effective vaccine, which drastically reduces the emergence of Ark-like viruses both at vaccination and after challenge.