Boyden-type chemotactic chambers were used to demonstrate that Mycoplasma gallisepticum (MG) was capable of migrating into chemotactic membranes. Scanning electron microscopy was used to confirm that MG could penetrate the membranes. To further demonstrate the invasive ability of MG, MG was deposited on the shell membranes of 9-day-old chicken embryos, and after 6 days of incubation, the presence of MG DNA in the allantoic fluids was detected by polymerase chain reactions. These results indicate that MG can penetrate cellular membrane, possibly by going through the porous cellular surface.