Population Dynamics of Bacteriophage and Bacillus Subtilis in Soil

Abstract

The dynamics of the interaction of populations of Bacillus subtilis and a temperate bacteriophage in soil microorganisms were examined. The purpose of the study was to investigate these dynamics in a structured habitat approaching that of their natural habitat in soil, in contrast to previous investigations in broth or chemostats. We addressed three main questions. What are the population dynamics of a phage—bacteria interaction in soil? What role might the heterogeneity of the soil play in shaping the interaction? Are the dynamics controlled more by the population biology of the phage or of the bacteria? The phage used was isolated from Arizona desert soil in which B. subtilis is common. Phage and bacteria were grown separately or together in sterile soil microcosms consisting of autoclaved soil rich in organic material. Densities of phage and bacteria were estimated through repeated sterile sampling of the microcosms by spreading precise dilutions of sampled soil suspensions onto plates of microbiological media. The plates for estimation of phage densities contained in addition a lawn of host bacteria. While the principal focus is on temperate phage ecology, the dynamics of a single example of interaction between B. subtilis and virulent phage is also presented. In all cases an initial epidemic of phage occurred, followed by stable equilibria lasting weeks to months. A threshold host density for phage outbreak occurs at °5 x 106 colony—forming units per gram of soil. At equilibrium the phage, both temperate and virulent, were much less abundant than the bacteria. The temperate phage did not depress the equilibrium host density, while the virulent phage lowered it by a factor of ten. The acidic soil of these experiments caused rapid and permanent inactivation of free phage, making the continuous interaction of phage and host essential for persistence of phage. Low levels of phage resistance (superimmunity or genetic) were typical of host populations. Most properties of the interaction between B. subtilis and phage in soil are quite different from those observed with chemostat populations of Escherichia coli and virulent phages. Potential doubling times and rates of increase in soil for phage and B. subtilis were calculated. At equilibrium, soil slows the interaction of phage and host relative to population growth in broth culture, and possibly also makes it heterogeneous in space and time. The life—history features of B. subtilis and temperate phage are considerably more complex than those of non—sporeforming bacteria and virulent phage. The biotic relationship of temperate phage and host may be distinct from predation or parasitism. Patterns in the ecology and evolution of temperate and virulent phage were explored, leading to the expectation that temperate forms will be more common among the phages of soil bacteria.