Production and Stabilization of Cells of Bacillus popilliae and Bacillus lentimorbus

Bacillus popilliae andB. lentimorbus grew most rapidly and to the greatest extent in aerated cultures at 30 to 32 C with oxygen absorption rates of 1 mmole of O2 per min per liter, or above. The control ofp H also increased the maximal populations attained. Media were developed which consistently produced cell populations of about 109 within 24 to 48 hr in aerated cultures of these two species. The acetic acid produced in highly aerated cultures was shown not to be responsible for the rapid loss of cell viability in stationary phase cultures. However, H2 O2 was very lethal to cells ofB. popilliae , and this species is known to have the capacity to produce it. Stationary-phase cells were partially stabilized by reducing the availability of oxygen after 24 hr of incubation on a shaker, and the addition of low levels of glucose further stabilized the cells. The most stable cells were those produced in a medium in which 4% Trypticase (BBL) and 0.1% barbituric acid were incorporated. A high percentage of these cells contained refractile bodies visible under a phase microscope. Although these bodies were not heat-resistant and lacked other characteristics of endospores, cells in cultures containing them had reasonably high viability for extended periods, as compared with those in control cultures.