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PubMed1961

Lysis of brucellae by the combined action of glycine and a lysozyme-like agent from rabbit monocytes.

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.

Animals↗
PubMed1962

Enzymatic properties of a phage-induced lysin affecting group A streptococci.

Doughty, C. C. (University of Illinois College of Medicine, Chicago) and James A. Hayashi. Enzymatic properties of a phage-induced lysin affecting group A streptococci. J. Bacteriol. 83:1058-1068. 1962.-Phage-induced lysis of group C streptococci releases into the medium a lysin which completely lyses group A streptococci. Partial purification of the lytic activity yields 47% of the original activity with a 17-fold purification. The activity was assayed by observing lysis of group A streptococci under standard conditions. The optimal pH range for lysis is from 6.0 to 6.7. A monovalent cation requirement satisfied by Na(+), K(+), or Li(+) is shown by the lysin. Lysis is stimulated by ethylenedi-aminetetraacetic acid (EDTA), chlortetracycline, streptomycin, and penicillin. It is inhibited by p-hydroxymercuribenzoate (pHMB), and the inhibition is reversed by cysteine. Other inhibitors include ristocetin A and specific antisera against the lysin. Isolated group A streptococcal cell walls are partially lysed by massive amounts of lysin. This partial lysis is not affected by EDTA, pHMB, chlortetracycline, streptomycin, or ristocetin A. It is concluded that the enzymatic process of lysis of isolated cell walls is not identical to the more complex process resulting in lysis of intact cells.

Bacteriolysis↗
PubMed1962

Lysis of Escherichia coli by sulfhydryl-binding reagents.

Schaechter, M. (College of Medicine, University of Florida, Gainesville) and K. Santomassino. Lysis of Escherichia coli by sulfhydryl-binding reagents. J. Bacteriol. 84:318-325. 1962-Washed suspensions of gram-negative rods were lysed by low concentrations of some sulfhydryl-binding and oxidizing reagents, but not by reducing agents. Some kinetic aspects of this phenomenon were studied with p-chloromercuribenzoate and Escherichia coli B/r. Structures resulting from the action of this reagent consisted of impure cell walls. These could be purified by treatment with trypsin. Cell walls prepared mechanically and cell membranes obtained by lysing protoplasts were not overtly affected by this chemical.

Bacteriolysis↗
PubMed1963

Mechanism of protection of cells by spermine against lysozyme-induced lysis.

Grossowicz, Nathan (Hebrew University Hadassah Medical School, Jerusalem, Israel) and Miriam Ariel. Mechanism of protection of cells by spermine against lysozyme-induced lysis. J. Bacteriol. 85:293-300. 1963.-Spermine (1 to 5 x 10(-3)m) was found to prevent lysis of Micrococcus lysodeikticus cells by lysozyme. This protection is due to the rapid and efficient stabilization by spermine of the protoplasts formed by lysozyme, and not to interference with the enzymatic activity. This conclusion concerning the spermine effect is based on chemical and microscopic evidence: (i) the amount of hexosamine, released from lysozyme-treated cells, was equal to or even higher in the presence of spermine than in its absence; (ii) microscopic and electron microscopic pictures showed clearly that the spermine-protected structures resembled protoplasts. On the other hand, the "spermine protoplasts" showed a high stability against hypotonic conditions in comparison with "sucrose protoplasts." The spermine-protoplast complex was very stable and could not be destroyed by water and various agents (NaCl, KCl, MgCl(2), cadaverine, and putrescine). A number of bases such as histamine, d-histidine, and streptomycin exhibited an effect similar to that of spermine, while diamines like cadaverine, putrescine, basic amino acids (lysine, arginine, l-histidine), as well as cations like magnesium, were virtually inactive.

Bacteriolysis↗
PubMed1963

CORRELATION OF SPECIATION WITH LYTIC RESPONSES OF THE ACHROMOBACTER.

Surdy, Theodore E. (Purdue University, Lafayette, Ind.) and S. E. Hartsell. Correlation of speciation with lytic responses of the Achromobacter. J. Bacteriol. 85:1011-1016. 1963.-Lysozymic lysis of six species of Achromobacter was investigated. Three of the six species were lysed with 33, 50, or 100 mug/ml of lysozyme; if higher concentrations of lysozyme were used, precipitation of cells occurred. "Insensitive" cells could be sensitized by the addition of potassium hydroxide, n-butanol, steapsin, or urea, as demonstrated by the subsequent addition of lysozyme. Not all species were sensitive to these agents in the same degree; hence, a spectrum was obtained after the use of the pretreating agents and lysozyme. Optimal clearing of suspensions was observed when cells were suspended in pH 6.6 physiological saline or 0.15 m phosphate buffer and incubated at 45 C. Heat treatment (75 C for 10 min) or freezing (-32 C) and thawing (room temp, 25 C) for one cycle did not increase the sensitivity of the cells to lysozyme. Injury to the cells was evident by the increased amount of lysis noted after pretreatment with potassium hydroxide. When cells were frozen and thawed for three cycles, four of the six species were sensitive to the action of lysozyme. Isolated cell walls elicited a similar lytic pattern to that of whole cells. Individuality of the lytic response of the species (from most sensitive to least sensitive-A. aquamarinus, A. butyri, A. viscosus, A. parvulus, A. guttatus, A. hartlebii) produced a separation scheme. Exhaustive tests proved it to be stable and reliable for these species. The organisms were identified, with the use of the separation scheme, by a person initially unfamiliar with the scheme or the culture.

Achromobacter↗
PubMed1963

STAPHYLOCOCCAL SENSITIZATION: SPECIFIC BIOLOGICAL EFFECTS OF PHAGE K ON THE BACTERIAL CELL WALL IN LYSIS-FROM-WITHOUT.

Ralston, Doris J. (University of California, Berkeley). Staphylococcal sensitization: specific biological effects of phage K on the bacterial cell wall in lysis-from-without. J. Bacteriol. 85:1185-1193. 1963.-Phage K, shown previously to sensitize staphylococcal-wall mucopeptide to the action of a phage-induced enzyme, virolysin, was found to act in a specific manner in that its sensitizing effects were restricted to chemical linkages affected by three staphylococcal lysins. These caused an immediate lysis, whereas egg-white lysozyme, which could also digest the wall mucopeptide, exerted variable effects, even when in the absence of phage it produced some lysis. Evidence was presented that the K(1) normal cell autolysin and the K phage virolysin could act synergistically with lysozyme on phage-sensitized cells, and that any effects observed with lysozyme were due to the simultaneous presence of trace amounts of these staphylococcal lysins. None of a series of lysozymelike agents from sea urchins, marine sepunculids, and from rabbit peritoneal histiocytes caused accelerated lysis of phage-sensitized cells, although like lysozyme they showed a slow lysis of phage-free living cells. Other enzymes which did not reduce the turbidity of sensitized cells included agents specific for intracellular components (proteins, lipids, nucleic acids), and enzymes, as decarboxylase, alkaline phosphatase, d-amino oxidase, and hyaluronidase. These results suggested that the main effects of the phage in sensitization were limited to areas of the cell wall involved in protection against the action of the staphylococcal lysins.

Animals↗
PubMed1963

MUTANT OF LAMBDA BACTERIOPHAGE PRODUCING A THERMOLABILE ENDOLYSIN.

Campbell, Allan (University of Rochester, Rochester, N.Y.) and Alice del Campillo-Campbell. Mutant of lambda bacteriophage producing a thermolabile endolysin. J. Bacteriol. 85:1202-1207. 1963.-Endolysin from lambda bacteriophage and a temperature-sensitive mutant thereof was partially purified. The mutant enzyme was distinguishable from the wild type by its greater rate of inactivation by high temperature and by urea. Lysogenic cells carrying the mutant phage did not lyse after induction if kept at 43 C, but, at times around 70 min after induction, rapid lysis occurred following transfer to lower temperatures. This lysis was not inhibited by cyanide or chloramphenicol and therefore probably resulted from enzyme already synthesized at the high temperature. Addition of these inhibitors to the culture at 43 C rapidly destroyed the ability to lyse after a subsequent temperature shift.

Bacteriolysis↗
PubMed1963

QUANTITATIVE STUDY OF ENDOLYSIN SYNTHESIS DURING REPRODUCTION OF LAMBDA PHAGES.

Groman, Neal B. (University of Washington, Seattle) and Grace Suzuki. Quantitative study of endolysin synthesis during reproduction of lambda phages. J. Bacteriol. 86:187-194. 1963.-Endolysin is presumed to be a phage-induced enzyme participating in lysis through its destructive action on the host cell wall. A method for assaying endolysin is described, which was utilized in studying endolysin synthesis at 37 and 44 C by induced strains of K-12 (lambda), K-12 (lambdatem), and K-12 (lambda112). In all cases, endolysin was detected prior to the appearance of mature, intracellular phage and was detected earlier at 44 C than at 37 C. It was synthesized at a linear rate, as was phage, and both syntheses terminated at the same time. Surprisingly, endolysin also accumulated under conditions in which induced K-12 (lambda112) exhibited lysis inhibition. Under these conditions, endolysin concentration per induced cell was 2 to 2.5 times that produced by normally lysing K-12 (lambda). Since alterations introduced into the lytic process by temperature, mutation, or both correlate well with the timing and rate of endolysin synthesis, the data tend to support the concept that endolysin determines the kinetics of the process. However, the accumulation of endolysin during lysis inhibition suggests the need for alternative hypotheses. One hypothesis is that although endolysin action is the key to lysis some preliminary steps are required to release the enzyme so that it may contact its substrate in the cell wall. A second hypothesis is that basically the lytic process involves an alteration in the permeability barrier of the cell and that lytic enzymes such as endolysin have evolved as an auxillary but dispensable mechanism to this process.

Bacteriolysis↗
PubMed1963

SPONTANEOUS LYSIS AND PHAGE-CARRIER STATE IN BRUCELLA CULTURES.

Renoux, Gerard (Université de Montpellier, Montpellier, France), and Andree Suire. Spontaneous lysis and phage-carrier state in Brucella cultures. J. Bacteriol. 86:642-647. 1963.-When broth or saline suspensions of 60 randomly chosen Brucella strains were directly poured onto plates of Albimi Agar, distinct plaques, indicative of phage activity, developed. Unselected Brucella cultures containing cell types that gave rise to several morphologically distinct colonial types all proved to be naturally phage-infected. Selection and study of some of these colonial types led to the following conclusions: (i) S or SI colonies do not carry the phage and are sensitive to it; (ii) pure R colonies are phage-resistant and do not carry the phage; (iii) butyrous or sticky white P ("Porteuses") colonies develop from "carrier cells" resistant to phage; and (iv) the progeny of cells of the P colony type segregate into cells that give rise to P, S, or SI colonies. However, when plates were streaked with a cotton swab soaked in the Brucella suspension, no visible plaque developed. The phenotypic changes occurring after phage infection are believed to play a role also under natural conditions; they are able to explain most of the natural behavior of Brucella. Their occurrence, however, does not exclude other genetic mechanisms that may produce similar phenotypic effects.

Agar↗
PubMed1963

SURVEY OF THE BACTERIOCINES OF ENTEROCOCCI.

Brock, Thomas D. (Indiana University, Bloomington), Barbara Peacher, and Deborah Pierson. A survey of the bacteriocines of enterococci. J. Bacteriol. 86:702-707. 1963.-A survey has been made of bacteriocine production by a wide variety of well-characterized strains of group D streptococci. On the basis of spectrums and sensitivity to chloroform, heat, and proteolytic enzymes, five distinct bacteriocines can be defined. Type 1 is produced by all Streptococcus zymogenes (S. faecalis var. zymogenes) strains, is active against a wide variety of gram-positive bacteria, and is also a hemolysin. Type 2 is produced by some S. liquefaciens (S. faecalis var. liquefaciens) strains, and acts on many enterococci as well as on certain other lactic acid bacteria. Type 3 is produced by certain strains of both S. faecalis and S. faecium, and inhibits a wide variety of group D streptococci, but is inactive against all other lactic acid bacteria tested except Leuconostoc citrovorum. Type 4 is produced by certain S. faecium strains and resembles in certain ways the type 3 activity, but differs from it in other ways. Type 5 has been found to be produced by only one proteolytic strain of S. zymogenes, and this bacteriocine has a very narrow spectrum. The strain that produces this bacteriocine also produces type 1 activity. No strain is sensitive to a bacteriocine of the type it produces.

Bacillus cereus↗
PubMed1963

PROBABLE IDENTITY OF A GROUP D HEMOLYSIN WITH A BACTERIOCINE.

Brock, Thomas D. (Indiana University, Bloomington) and Joseph M. Davie. Probable identity of a group D hemolysin with a bacteriocine. J. Bacteriol. 86:708-712. 1963.-All strains of Streptococcus zymogenes (S. faecalis var. zymogenes) produce a bacteriocine which is active against lactic acid bacteria and most other grampositive bacteria. Mutants which have lost the hemolytic characteristic lose at the same time their bacteriocine-producing ability. A strain which was resistant to the bacteriocine but which was nonhemolytic and nonbacteriocinogenic was irradiated, and two hemolytic mutants were isolated from it. These mutants were also bacteriocinogenic. Thus, the two activities are gained or lost together by mutation. Both activities are destroyed by chloroform vapors and are antagonized by lecithin. Both activities are destroyed at the same rate by treatment at 45 C under mildly acid conditions, and both activities are stable when heated in agar. The two activities are produced in parallel during the growth cycle, and disappear in parallel. The possible ecological role of a substance which is both a hemolysin and a bacteriocine is discussed.

Bacteriolysis↗
PubMed1964

ISOLATION AND CHARACTERIZATION OF BACTERIOPHAGES ACTIVE ON CLOSTRIDIUM SPOROGENES.

Betz, John V. (St. Bonaventure University, St. Bonaventure, N.Y.), and Kenneth E. Anderson. Isolation and characterization of bacteriophages active on Clostridium sporogenes. J. Bacteriol. 87:408-415. 1964.-Twelve bacteriophages active on the anaerobic species Clostridium sporogenes were studied. Four of these were isolated by the authors, and eight were obtained from L. S. McClung of Indiana University. The 12 phages studied could be distinguished into three groups on the basis of their plaque morphology, host range, receptor sites, and serological relationships. One group contained ten of the phages which were serologically related. These were separated into three subgroups on the basis of plaque morphology, host range, and receptor sites. The heat sensitivities of four phages were correlated with their classification in this scheme. Of 25 strains of C. sporogenes tested for lysogenicity, none was found to be lysogenic, but 9 produced bacteriocin-like substances and 20 were sensitive to one or more of these.

Bacteriolysis↗
PubMed1964

DISTRIBUTION OF RADIOACTIVITY IN AUTOLYZED CELL WALL OF BACILLUS CEREUS DURING SPHEROPLAST FORMATION.

Kronish, Donald P. (Warner-Lambert Research Institute, Morris Plains, N.J.), Raam R. Mohan, and Benjamin S. Schwartz. Distribution of radioactivity in autolyzed cell wall of Bacillus cereus during spheroplast formation. J. Bacteriol. 87:581-587. 1964.-Spheroplasts of Bacillus cereus strain T were produced from cells grown in the presence of uniformly labeled C(14)-glucose. At regular intervals during spheroplast formation, enzymatically degraded cell wall was isolated by a new procedure. Radioactivity of solubilized cell wall in cell-free material increased from 2.5 to 42% of the total incorporated label during spheroplast formation. The rate of cell-wall degradation as measured by increase in radioactivity was biphasic with relative slopes of 2.0 and 5.0. During autolytic depolymerization of B. cereus cell wall, two major components were solubilized at different rates. Chemical fractionation revealed these to be a peptide and a mucopeptide. The possibility of two enzymes being involved in spheroplast formation and cell-wall degradation is discussed.

Amino Acids↗
PubMed1964

"CELL-WALL PROTEIN A" OF PSEUDOMONAS AERUGINOSA AND ITS RELATIONSHIP TO "ORIGINAL ENDOTOXIN PROTEIN".

Homma, J. Yuzuru (University of Tokyo, Tokyo, Japan), and Nachiko Suzuki. "Cell-wall protein A" of Pseudomonas aeruginosa and its relationship to "original endotoxin protein." J. Bacteriol. 87:630-640. 1964.-To compare the properties of two kinds of proteins, one obtained from the cell wall of Pseudomonas aeruginosa and the other from the endotoxin in the autolysate, the proteins were purified to such an extent that they were proved to be homogeneous by zone electrophoretic and ultracentrifugal analyses. Although there were some differences between the two in such values as nitrogen and phosphorus contents and sedimentation constants, their protein portions were found similar to each other in the ratio of amino acid compositions. It was proved serologically that the two proteins possessed a common specific antigen. They were found to be the same in their potencies in eliciting the Shwartzman phenomenon and pyrogenic reaction. The results of pyocine tests with various sensitive strains revealed that, against the sensitive strains, the spectra of both proteins were almost the same. Their pyocine activities were destroyed through digestion by protease, trypsin, and Nagarse. Either of the rabbit antisera against the two proteins could neutralize pyocine activities of both proteins. Serum-absorption tests proved that the pyocine-neutralizing antibodies of both antisera could be completely absorbed with each of the proteins. Under appropriate conditions, their pyocine activities were masked partially in vitro by the lipopolysaccharde of the endotoxin.

Amino Acids↗