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Introduction and Objectives: A new interaction between avian influenza virus (AIV) and infectious bronchitis virus (IBV) replication interference was identified in this work using chicks by ELISA antibody, quantitative reverse transcription-polymerase chain reaction (qRT-PCR), cytokine analyses and histochemistry. Materiel and Methods: In this study chicks were infected with IBV vaccine strain (H120) and a H9N2 LPAIV (A/CK/TUN/145/2012) simultaneously or sequentially three days apart. Our research studied first the eventual replication interference of AIV and IBV in vivo using qualitative and quantitative real-time reverse transcription-polymerase chain reaction (real-time qRT-PCR) to accurately detect either AIV and IBV genomes or the number of viral copies during mixed infections. Second, we determined the amount of IL-1 beta by ELISA and antibody of the infected chick's sera with AIV alone, IBV alone, Mixed AIV + IBV, IBV 3 days AIV and AIV 3 days IBV. Our data revealed that the second virus inoculated inhibits the growth of the first one independently of its nature. Results and Conclusion: In conclusion, co-infection of chicks with AIV and IBV affected the replication dynamics of these viruses and affect clinical signs. These results propose that infection with heterologous virus may result in temporary competition for cell receptors or competent cells for replication, most likely interferon-mediated. In conclusion, the second virus inoculated inhibits the first one by different mechanisms. [ABSTRACT FROM AUTHOR]