Serological Studies of the Multiplication of a Staphylococcal Bacteriophage

Abstract

In a previous paper (Rountree, 1951a) it was reported that complement fixation occurred wben coli bacteriophase T5 reacted with its homologous antiserum in the presence of complement. By cyanide lysis during the latent period. T5 infected cells were shown to synthesize antigenic material reacting iu tin* fomplcnient fixation test at a stage in the phage growtli cycle at which few or no infective phage particles could be demonstrated. It was postulated that the final step in phage multiplication, i.e. the production of complete infective partieies takes place in the last part of the latent period after a pool of tbis antigenically reacting material has been accumulated.In the present paper an account is given of the production of antipbage sera ill rabbits and of the use of these sera in studying the phenomena of pbage adsorption and multiplication. A staphylococcal phage, .3A of Wilsou and Atkinson's (1945) series of typing phages. and its host strain, 284. have been used since it was desired to see if results comparable with those obtained with the T5-eoli system could be obtained witb a different phage-host system. No evidence of lysogenicity in strain 284 was obtained in a previous study (Rountree. 1949a). The presence of lysogenicity cannot, however, be excluded since a suitable indicator strain for a carried phage may not be available. METHODS.Phnge eonnis were mnde hy the surface agar plating method, each plate accommodating 4 inopul.-i of 0-01 ml. each.Phnge adanrption wns measured bv titiation of free ph.ige in the supernatants of infented broth cultures centriftiged at various timea after addition of the phage.Phage neutraiizing antibody wns titrated by the method of Burnet, Keogh and Laah (1937).

Infection by Bacteriophage T5 and Its Intracellular Growth—a Study by Complement Fixation

“…There is no evidence of inhibition of fixation in excess serum (at constant antigen). Similar results have been obtained with T2:anti-T2 (Lanni and Lanni, unpublished data), but Rountree (1951Rountree ( , 1952) has failed to observe antigen-excess inhibition with either phage T5 or phage 3A. (2) With serum diluted 1:800 and less so at 1:400, the curve shows an aberration at high antigen concentration due to anticomplementary effect of the antigen. At the extreme right, all curves approach basal levels set by the anticomplementary effect of serum at various dilutions.…”

“…There is, therefore, no evidence of an early completion of a pool of antigen later incorporated into phage (see Rountree, 1951 figure 2 suggested that the infectivity but not the serological activity of the infecting phage is lost shortly after adsorption. Since, however, the infected cultures were lysed artificially, the experiments gave no evidence as to whether the antigen entered the host cell as suggested by Rountree (1951Rountree ( , 1952 remained outside as suggested by findings of Hershey and Chase (1952) with T2.…”

“…(3) The complement-fixing activity of the premature lysates (curve C) shows a nearly constant period of about 18 minutes, during which the activity is almost identical with the total input activity (arrow 2). There is no evidence that the input antigen disappears during infection (see Rountree, 1951Rountree, , 1952.…”

Lysogeny in Staphylococci

“…Supernatants of the broth cultures or of broth washings from the agar plates were filtered through Seitz E. K. filter pads. The serological group) of each phage was determined by methods de-scribed by Rountree (1949), using specific antisera prepared by the immunization of rabbits to phages of the serological groups A, B, F, and L;…”

Variations in a Related Series of Staphylococcal Bacteriophages

“…When propagated in staphylococcus strain Pa, the two phages displayed differences in plaque size, adsorption constants in broth (Rountree, 1952), requirements of divalent cations for penetration and growth (Rountree, 1955), length of the minimum latent period and average burst size ( Table 2). Their ability to show plaques on a number of indicator strains of staphylococci is given in Table 3 and expressed as: ( a ) numbers of plaqueslml.…”