Mycoplasmosisis one of the most important diseases in the poultry industry. Mycoplasma is the causative agent of this disease which have various species.Two species;Mycoplasma gallisepticum(MG) and Mycoplasma synoviae (MS) are the most important species due to the losses in the production farms of industrial poultry achieving rapid, accurate and definite diagnosis of Mycoplasma has an abundant importance because early and definite diagnosis can protect hygiene farm management on keeping herd health. In many countries such as Iran, the disease and it’s complications have still remained as a serious problem. Given this issue, we decided to identify the Mycoplasma infection from broiler poultry flocks through culture method.150 carcasses of broiler chicken belonging to 50 broiler flocks were sampled in which the conflict between air sacs and secretions in the airways, trachea and bronchi were seen. The samples were taken from trachea, palatine cleft, nasal passages and air sacs. Then, the samples were cultivated into PPLO liquid medium by the use of membrane filters(0.45 micron) . All of the media were incubated at37 ° C they were examined for pH (color) changes every 48 hours. During the first 24 hours after cultivation, every color change to yellow or dark visible with the naked eye was considered as bacterial contamination, therefore, the contaminated samples were removed from the incubator. The color change in the liquid media was compared with the uninoculated medium as negative control. If a color change was observed in the liquid media after 48h, subculture was done in the PPLO agar. The plates were incubated at 37 °C for 14 days. They were examined for Mycoplasma colonies under a microscope with magnification of 10 in every other day. The results showed that from 150samples obtained from 50 broiler flocks, 16 (10.66%) were positive for Mycoplasma ,4 flocks (8%) of 50 studied flocks in terms of contamination were positive for Mycoplasma. The positive cultures were finally confirmed by using PCR method with universal primer.

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