Background: Mycoplasma is one of the most important pathogens of respiratory system in poultry. OBJECTIVES: The aim of the present study was to identify Mycoplasma spp. isolated from pigeons. METHODS: Sixteen pooled samples were provided and cultured on PPLO medium and finally the DNA was extracted from the resulting single colonies. RESULTS: Through 16S rRNA gene amplification Mycoplasma genus has been detected. Overall, 31% (5 out of 16) of pooled samples were positive which were identified as Mycoplasma cloumborale and Mycoplasma gallinaceum. CONCLUSIONS: Screening of large numbers of pigeons for known poultry pathogenic mycoplasmas will be required to establish the role of pigeons in the spread and maintenance of these organisms in the environment.