STUDIES ON THE BACTERIOPHAGE OF D'HERELLE : VIII. THE MECHANISM OF LYSIS OF DEAD BACTERIA IN THE PRESENCE OF BACTERIOPHAGE

Abstract

We have been able to confirm the observations of Twort as well as of Gratia, that dead staphylococcus may undergo lysis if, in addition to a suitable bacteriophage, there is also present live staphylococcus. Moreover, we have endeavored to ascertain the mechanism of this phenomenon and have found that in order to elicit it it is necessary to control the numbers of live and dead bacteria in the mixture. An excess of dead bacteria interferes with lysis by adsorbing the bacteriophage before it has the opportunity to initiate necessary changes in the live bacteria, so that all lysis is prevented. The phenomenon is specific, that is, the lysis of live bacteria is accompanied by lysis of dead bacteria of the same species only. Lysis of dead bacteria occurs best with staphylococcus, an organism which easily undergoes spontaneous autolysis under appropriate conditions. In the case of B. coli or B. dysenteriae the lysis of the dead bacteria is uncertain. Dead bacteria need not be present in the mixture at the beginning of the experiment; they will be dissolved if added any time before, during, or after the completion of lysis of live bacteria. If the test is performed so that a suitable semipermeable membrane is interposed between the dead and live bacteria, the dead bacteria are not dissolved, in spite of the lysis of live bacteria on the other side of the membrane. The agent determining the lysis of dead bacteria is not diffusible, while the principle initiating the lysis of live bacteria diffuses freely and is demonstrably present on both sides of the membrane. The complete independence of the agent causing dissolution of dead bacteria from bacteriophage can also be shown by separating the two agents by means of filtration, or by adsorption on bacteria. The ferment-like substance responsible for the lysis of dead bacteria is different from the bacteriophage. It is not diffusible through collodion, it is easily adsorbed on clay filters, it is heat-labile, and is inactivated on standing. An agent possessing identical properties was found in cultures of staphylococcus undergoing spontaneous autolysis in the absence of bacteriophage, but in this instance the agent appeared in the filtrates considerably later than it did when phage was present.