Biological evaluation of Mycoplasma pulmonis temperature-sensitive mutants for use as possible rodent vaccines

Temperature-sensitive mutants (TSMs) of Mycoplasma pulmonis were produced by treating the wild-type strain with N-methyl-N'-nitro-N-nitrosoguanidine. Three TSms were selected at 38°C, as a restrictive temperature, and at 34°C, as a permissive temperature. Two TSMs, UTCMI and UTCMII, were proven to be nonpathogenic but immunogenic. In addition, they did not induce pneumonia, tracheitis, or tympanitis but did induce mild rhinitis. They were stable after 10 passages in vitro and in vivo. They elicited excellent antibody production and cell-mediated immunity in vaccinated rats. They also were not mitogenic to rat lymphocytes. Rats immunized intranasally with these TSMs were significantly protected against challenge with wild-type organisms. These mutants were morphologically and serologically indistinguishable from the wild-type organisms. The growth characteristics and antibiotic sensitivities were similar to those of wild-type organisms, except that they grew only at 34°C. In contrast to wild-type organisms, they did not bind to or lyse sheep erythrocytes. Thus, these TSMs may qualify as a vaccine to prevent M. pulmonis infection in rats.