A NEW PROPERTY OF PHAGE GROUP II STAPHYLOCOCCUS AUREUS STRAINS: HOST RESTRICTION OF PHAGE K14.
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Biomedical subjects
Publications and source records attributed to B S BAER.
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Ralston, D. J. (University of California, Berkeley) and B. S. Baer. Inhibitory action of phage K on staphylococcal dehydrogenases. I. Effect on various strains of Staphylococcus aureus, including members of the phage-typing series. J. Bacteriol. 86:666-672. 1963.-The polyvalent phage K was found to depress the dehydrogenase activities of a large number of Staphylococcus aureus strains, as measured by the reduction of triphenyl tetrazolium chloride to the insoluble red formazan. The inhibition occurred immediately after the absorption of a multiplicity of phage particles, and was independent of the infectibility of the strains and of the killing ability of the phage. It appeared to be closely associated with the phenomenon of sensitization-a change in the cell surface which increased the susceptibility of the wall to digestion by soluble staphylococcal lysins and simultaneously abolished the capacity to synthesize phage. The inhibitory effect occurred both in nutrient media and in a nongrowth glucose - phosphate buffer supplemented with cysteine.
Ralston, Doris J. (University of California, Berkeley), B. S. Baer, and S. S. Elberg. Lysis of brucellae by the combined action of glycine and a lysozyme-like agent from rabbit monocytes. J. Bacteriol. 82:342-353. 1961.-An acid-extractable lytic material was obtained from rabbit monocytes. It acts on a substrate in the walls of brucellae and has properties similar to egg-white lysozyme. Brucella melitensis strain Rev Is is completely resistant to this agent and also to crystalline lysozyme, but when the cells are exposed to sufficient amounts of glycine, the surface is rendered susceptible to these lytic agents. Rough type Rev Is cells are more susceptible than smooth, and the virulent B. melitensis strain 6015 is most resistant.
LYSIS FROM WITHOUT (LFW) OCCURS IN TWO STEPS: (1) sensitization of cells by phage, which renders the cells susceptible to (2) destruction of an essential cell structure by an extracellular lytic enzyme. Virolysin, from phage-infected cells, was used in these studies. Normal cell autolysin is also effective. Evidence is presented that: 1. Neither phage nor lysin alone causes LFW. 2. Sensitization requires phage adsorption. 3. It can be caused by non-infectious particles. This establishes a new biological activity of the particle. 4. Heat, U.V., detergents, penicillin, and other damaging agents also sensitize cells. 5. Sensitization involves a non-lethal, reversible reaction. 6. Sensitization by phage prevents virus synthesis. Following adsorption, a cell can undergo sensitization or infection but not simultaneously. When only a few particles are adsorbed, infection can occur; when sufficient particles are adsorbed, sensitization takes place. 7. Quantitative aspects of LFW are described. Lysis proceeds logarithmically. The lysis end-point depends upon the phage concentration but is independent of the enzyme concentration.
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There were differences in the way the lysogenic strain N of B. mycoides and the parent indicator strain grew on nutrient agar and in nutrient broth. 1. On agar, the indicator culture traveled more quickly over the agar surface than the phage-carrying strain; the total extent of spread was greater. 2. In broth, the indicator strain grew diffusely throughout the liquid, the lysogenic cells in clumps. The virus-infected strain appeared to grow more slowly. This may reflect (a) the effect of aggregation on the generation time of the lysogenic strain, (b) an active lytic process in the lysogenic population which is further enhanced by the effect of clump formation on the environment of the cell.
In nutrient broth at 30 to 32 degrees C., the cycle of virus growth (following adsorption) in lag phase cells of B. mycoides N included a period of intracellular multiplication, ranging from 0.8 to 1.3 hours, succeeded by a sharp rise in the free phage titer and then by a slower rise or a plateau in the extracellular phage content. The yield of virus per infected cell at 30 degrees C., as determined by a modified Burnet dilution technique, was about 76 plaque-forming particles. During the latent period, multiply infected cells showed no change in numbers. Coinciding with phage release, incomplete clearing occurred. The unlysed, remaining cells multiplied and the turbidity rose again. These survivors and their progeny were lysogenic.
Experiments were performed to determine the mechanism of release of phage from the lysogenic strain of B. mycoides N. The results suggest that qualitatively the same situation obtains as in the phage-carrying cultures of B. megatherium 899 and E. coli Li; i.e., the population consists of two kinds of cells: "lysogènes potentiels" and "producteurs." Quantitatively, however, there are more "producers" in a broth culture of the lysogenic B. mycoides N, at least curing the first 4 to 8 hours after cells have been suspended in fresh medium, suggesting that the interaction between host and parasite is one in which the balance is easily swung in favor of the virus. These conclusions are based upon the following lines of evidence: (1) the slow "growth rate" of the lysogenic culture, (2) the fact that the colony count falls far below the plaque count or the filament count (which correspond) for a well washed suspension, (3) the increase in phage output in a large number of tubes, each containing a small number of lysogenic cells, after a few hours' incubation in nutrient broth at 30 degrees C.