Use of polymerase chain reactions for the detection of M. gallisepticum, M. imitans, M. iowae, M. meleagridis and M. synoviae in birds of prey

Mycoplasmas are pathogens of different avian species, but the role of Mycoplasma in raptors is not yet completely determined. In the past Mycoplasma species known to be pathogenic for poultry were described in raptors but do not seem to occur regular. Therefore species-specific PCRs for the detection of M. gallisepticum, M. imitans, M. iowae, M. meleagridis and M. synoviae were evaluated for the use in non- poultry species. The specificities of the PCR-methods were investigated using avian Mycoplasma strains, further Mycoplasma isolates and walled bacteria and found to be specific. The sensitivities of the different PCR- assays varied between 100 fg and 10 pg DNA. Fiftythree tracheal swabs from healthy captive and free ranging birds of prey were investigated using the above described PCRs. In none of the cases an amplicon was obtained using the PCRs for the detection of M. gallisepticum/ M. imitans, M. iowae and M. synoviae. Using species specific primers for the detection of M. meleagridis a product was amplified from eight samples. Restriction enzyme analysis as well as sequencing of PCR products demonstrated these results as false positive. Alignment studies of the sequenced products with the 16S rRNA of various Mycoplasma species in GenBank demonstrate an identity of 91% to M. meleagrids but of 98% to M. buteonis or M. gallopavonis. Isolation and differentiation of one of these mycoplasmas suggests this isolate as a possible new species.