The effect of an inactivated vaccine on the horizontal transmission of Mycoplasma gallisepticum was quantified in a transmission model. Twenty non-vaccinated and 20 vaccinated 23-week-old specific pathogen free hens were housed in pairs, while five individually housed hens acted as a negative control group. Each pair consisted of a challenged chicken (10(4) colony forming units intratracheally) and a non-challenged susceptible contact bird. Infection was monitored by serology, quantitative polymerase chain reaction and culture. All non-vaccinated and vaccinated in-contact chickens became infected with M. gallisepticum. The 95% confidence interval of the reproduction ratio, R (a measure of transmission defined as the average number of secondary cases caused by one infectious individual) was 4.48 to infinity in both groups. However, the logarithm of the area under the curve in the vaccinated group was 0.51 lower (P = 0.02) than in the non-vaccinated group, indicating that there was an effect of vaccination on the levels of potential shedding of M. gallisepticum. Nevertheless, the results of this study indicate that the use of an inactivated M. gallisepticum vaccine will not reduce the horizontal transmission of M. gallisepticum between laying hens.