By use of a simple device for continuous CO2 gassing of Ureaplasma urealyticum cultures growing in a liquid medium, we have been able to separate some of the effects of urea, CO2, ammonia, and pH on growth. The CO2 acted as a superior buffer in the pH range 5.7 to 6.8, which is optimal for Ureaplasma growth. It was, therefore, possible to observe the effect of repeated additions of urea to the culture without alkalinization of the growth medium. We found that the repeated additions of urea did not enhance Ureaplasma growth, and the resultant accumulation of ammonium ions (greater than 2,000 microng/ml) did not cause more rapid death under these conditions. By abruptly changing the gaseous environment from CO2 to N2, it was possible to cause a rapid pH change in the culture to a value above 8.0. This resulted in a more rapid death of the organisms.