CHANGES IN THE SHAPE AND SIZE OF BACTERIUM COLI AND BACILLUS MEGATHERIUM UNDER THE INFLUENCE OF BACTERIOPHAGE—A MOTION PHOTOMICROGRAPHIC ANALYSIS OF THE MECHANISM OF LYSIS

This paper contains the records of a motion photomicrographic investigation of the lysis of Bact. coli and B. megatherium by bacteriophage. The bacteria mixed with bacteriophage were grown on moist nutrient agar in small culture chambers on the stage of a microscope in an incubator maintained at 37°C. The apparatus used permitted continuous inspection of the preparations. Photographs were made at the rates of 2 and 30 per minute and at the rate of 8 per second during the terminal stage of lysis of Bact. coli. The accurately timed films were studied by rapid projection and by the projection of single frames. Measurements of dimensions of cells, calculations of volumes, information on generations, generation times and duration spans are presented in the tables. Similar information on normal cultures grown and photographed in the same way is furnished for comparison. Groups of serial photographs are reproduced in the plates to illustrate the special features observed. These observations seem to us to warrant the following conclusions: 1. Enlargement or swelling of the cells of Bact. coli usually, but not always, precedes lysis. Some of the enlargement is an expression of increase of cell substance and is not altogether due to imbibition of water. Cells of early generations of Bact. coli enlarge to greater absolute and relative proportions than cells of later generations. Enlargement does not occur before lysis in B. megatherium. 2. The terminal stage of lysis of Bact. coli is explosive, occupying ½ to ⅞ second. The terminal stage of lysis of B. megatherium is a slow disintegrative process, extending over 2–10 minutes. 3. Bacteriophage inhibits fission of some cells, but does not stop the reproduction of other cells in contact with it. The genealogical records of six generations of cells of Bact. coli and of two generations of cells of B. megatherium indicate that bacteriophage may be transmitted through parents to the offspring which ultimately undergo lysis. 4. Bacteriophage spreads by contact through a group of cells and also along paths determined by genetical relationships. 5. A large amount of cellular debris remains after the lysis of the cells in both of these species of bacteria. This residue of material is in the form of irregularly shaped masses and granules. This material is not in solution at the time of lysis and appears not to be digested or hydrolized. 6. Theories of the mechanism of lysis are discussed. It is suggested that reduction of surface tension of the cells may be an important factor in the mechanism of lysis.