Mycoplasma imitans (Mim) has been isolated from ducks, geese and partridges, and is closely related to Mycoplasma gallisepticum (Mg). The pathogenicity of Mim for chicks was investigated in single and mixed infections with infectious bronchitis virus (IBV) by giving IBV strain M41 at 1-day-old and Mim 2 days later. Single infections with IBV or Mim were also performed. No clinical signs or gross lesions were seen in chicks infected with Mim or uninfected control chicks, but they were seen in the other two groups. Clinical scores were consistently higher in birds with mixed infections than those infected with IBV alone, and were significantly higher (P < 0.05) between days 7 and 14. More birds developed sinusitis, tracheitis and airsacculitis (with greater severity) in the mixed than the single IBV infections. Mim was recovered more frequently and in greater numbers from the respiratory tract of birds with mixed than single infections. It was recovered from the lower trachea, air sacs and lungs only in mixed infections. Seroconversion to Mim occurred by day 14 in mixed infections, but not until day 21 in single infections. It appears that Mim can act synergistically with IBV in young chickens in a similar manner to Mg, although Mg may act as a primary pathogen under some circumstances.
The present study was based on the reverse transcription polymerase chain reaction (RT-PCR) of the 16S ribosomal nucleic acid (rRNA) of Mycoplasma for detection of viable Mycoplasma gallisepticum. To determine the stability of M. gallisepticum 16S rRNA in vitro, three inactivation methods were used and the suspensions were stored at different temperatures. The 16S rRNA of M. gallisepticum was detected up to approximately 20-25 h at 37 °C, 22-25 h at 16 °C, and 23-27 h at 4 °C. The test, therefore, could detect viable or recently dead M. gallisepticum (< 20 h). The RT-PCR method was applied during an in vivo study of drug efficacy under experimental conditions, where commercial broiler-breeder eggs were inoculated with M. gallisepticum into the yolk. Hatched chicks that had been inoculated in ovo were treated with Macrolide 1. The method was then applied in a flock of day 0 chicks with naturally acquired vertical transmission of M. gallisepticum, treated with Macrolide 2. Swabs of the respiratory tract were obtained for PCR and RT-PCR evaluations to determine the viability of M. gallisepticum. This study proved that the combination of both PCR and RT-PCR enables detection and differentiation of viable from non-viable M. gallisepticum.