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PubMed1963

EFFECT OF ANAEROBIC GROWTH ON NITRATE REDUCTION BY STAPHYLOCOCCUS EPIDERMIDIS.

Jacobs, N. J. (American Meat Institute Foundation, Chicago, Ill.), J. Johantges, and R. H. Deibel. Effect of anaerobic growth on nitrate reduction by Staphylococcus epidermidis. J. Bacteriol. 85:782-787. 1963.-In anaerobic cultures, a strain of Staphylococcus epidermidis failed to reduce nitrate, although nitrate was reduced in stationary cultures grown in the presence of air. Resting suspensions of cells grown in air reduced nitrate. Although suspensions of cells grown anaerobically did not reduce nitrate, the nitrate reductase enzyme was shown to be present. When the artificial electron carrier, benzyl viologen, was added to suspensions of cells grown anaerobically, nitrate was reduced, indicating that an electron carrier required for nitrate reduction was missing. The addition of hemin to the growth medium enabled anaerobic cultures to reduce nitrate.

Benzyl Viologen↗

PubMed1963

COMPARATIVE PHYSIOLOGY OF ANTIBIOTIC-RESISTANT STRAINS OF STAPHYLOCOCCUS AUREUS.

Brown, Ruby L. (North Carolina State College, Raleigh) and James B. Evans. Comparative physiology of antibiotic-resistant strains of Staphylococcus aureus. J. Bacteriol. 85:1409-1412. 1963.-A collection of antibiotic-resistant strains of Staphylococcus aureus isolated from clinical sources was studied with respect to nutritional requirements and common diagnostic tests. Contrary to numerous reports in the literature indicating changes in these characteristics in antibiotic-resistant mutants, the present cultures were typical members of the taxonomic species S. aureus. They were coagulase-positive, fermented both glucose and mannitol under anaerobic conditions, produced acetoin from glucose, grew and produced black colonies on tellurite glycine agar, required both thiamine and nicotinic acid, and did not require other vitamins or purines. It is suggested that in most instances these cultures from clinical sources represent spontaneous mutants having genetic changes limited largely to loci concerned with antibiotic resistance. Most reports of extensive changes in physiology and nutritive requirements by antibiotic-resistant strains of S. aureus are based on studies of resistant strains selected after exposing a large population of the parent sensitive strain to toxic levels of antibiotics, chemical mutagens, or irradiation. Such isolates may have widespread genetic damage at other loci in addition to those concerned with their antibiotic resistance.

Anti-Bacterial Agents↗

PubMed1963

COAGULASE-NEGATIVE MUTANTS OF STAPHYLOCOCCUS AUREUS: GENETIC STUDIES.

Korman, Ruth Z. (Cornell University, Ithaca, N.Y.). Coagulase-negative mutants of Staphylococcus aureus: genetic studies. J. Bacteriol. 86:363-369. 1963.-The behavior in mutation and transduction of pleiotropic coagulase-negative mutants of Staphylococcus aureus PS 53 (NCTC 8511) was analyzed. Coagulase-positive colonies were recovered, as well as a novel phenotype resistant to some cell-wall inhibitors and differing in sugar fermentation pattern. The hypothesis that the coagulase-negative strains differ from the original propagating strain in the structure or organization of the cell wall is discussed.

Carbohydrates↗

PubMed1963

HEMOLYSIN-INHIBITING SUBSTANCE IN STAPHYLOCOCCUS AUREUS STRAINS.

Cetin, E. T. (Istanbul University, Istanbul, Turkey). Hemolysin-inhibiting substance in Staphylococcus aureus strains. J. Bacteriol. 86:407-413. 1963.-Of 144 Staphylococcus aureus strains isolated from pathological specimens, 3.4% did not cause hemolysis on sheep blood agar; the remainder produced hemolytic and semihemolytic zones, most of which were surrounded by a dense red band. Most of the strains causing pronounced hemolysis and a large dense red band on sheep blood agar also produced a dense red band on human blood agar after incubation at 37 C for about 1 week. In the dense red band on human blood agar, circles of hemolysis were observed when petri dishes were kept at room temperature for approximately 1 more week. The dense red band inhibited delta hemolysis of some of the S. aureus strains growing nearby. Certain strains failing to produce the dense red band on human blood agar inhibited delta hemolysis of other strains grown near them. A hemolysis-inhibiting substance (HIS) was produced in broth and agar media, and could be extracted from agar cultures. HIS was stable for 1 hr at 56 C and for 15 min at 100 C. It lost 75% of its effect when heated for 30 min at 100 C and became ineffective when heated at the same temperature for 45 min. Conditioned hemolysis occurred when some saprophytic gram-positive bacteria grew near or within the dense red band. These organisms also produced conditioned hemolysis without the presence of a visible dense red band, when growing near the strains inhibiting delta hemolysis. When such strains were grown on human blood agar, a conditioned hemolysis also occurred after the border of the medium was flamed.

Agar↗

PubMed1963

PYRITHIAMINE ADAPTATION OF STAPHYLOCOCCUS AUREUS. II. TRICARBOXYLIC ACID CYCLE AND RELATED ENZYMES.

Das, S. K. (Calcutta University, Calcutta, India), and G. C. Chatterjee. Pyrithiamine adaptation of Staphylococcus aureus. II. Tricarboxylic acid cycle and related enzymes. J. Bacteriol. 86:1157-1164. 1963.-Evidence for the stimulated operation of the tricarboxylic acid cycle in Staphylococcus aureus after pyrithiamine adaptation is presented. In the cell-free extracts, isocitric, glutamic, malic, and succinic dehydrogenases and catalase were found to be stimulated after the adaptation of S. aureus to pyrithiamine. Besides such stimulation, the appearance of isocitratase and malate synthetase in the adapted strain supports the appearance of the glyoxalate bypass after such adaptation. There is little change in the activities of reduced nicotinamide adenine dinucleotide (NADH) and reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidases and diaphorase. Lactic dehydrogenase, NADH-cytochrome c reductase, and NADPH-cytochrome c reductase could not be demonstrated either in the normal or in the pyrithiamine-adapted S. aureus. These observations support the postulation that there is a stimulation in the tricarboxylic acid cycle and can account for the very marked stimulation in the utilization of acetate by the organism after adaptation.

Acclimatization↗

PubMed1964

IN VITRO PHAGOCYTOSIS OF STAPHYLOCOCCUS AUREUS WITH SPECIAL REFERENCE TO A POSSIBLE ANTIPHAGOCYTIC FUNCTION OF COAGULASE.

Borchardt, Kenneth A. (Tulane University School of Medicine, New Orleans, La.), and William A. Pierce, Jr. In vitro phagocytosis of Staphylococcus aureus with special reference to a possible antiphagocytic function of coagulase. J. Bacteriol. 87:311-315. 1964.-Rabbit antisera were prepared to Staphylococcus aureus strains of bacteriophage types 81 and 42D with vaccines consisting of washed, heat-killed cells grown in media which either did or did not support coagulase formation. Partially purified coagulases were prepared, and were used to produce rabbit anticoagulase antisera. In vitro phagocytic studies, with the two staphylococcal strains plus rabbit polymorphonuclear peritoneal exudate cells and the various types of rabbit antisera, did not yield evidence that coagulase per se contributes to staphylococcal resistance to either ingestion or intracellular killing. The role of coagulase, if any, in the virulence of these organisms for rabbits (and probably for man) must therefore be attributed to some other mechanism.

Animals↗

PubMed1964

FORMATION OF "PROTOPLASTS" OF STAPHYLOCOCCUS AUREUS WITH A FUNGAL N-ACETYLHEXOSAMINIDASE.

Hash, John H. (Lederle Laboratories, Pearl River, N.Y.), Marcia Wishnick, and Philip A. Miller. Formation of "protoplasts" of Staphylococcus aureus with a fungal N-acetylhexosaminidase. J. Bacteriol. 87:432-437. 1964.-"Protoplasts" of Staphylococcus aureus were obtained when cells were treated with a fungal N-acetyl-hexosaminidase in hypertonic sucrose solution. The "protoplasts" were sensitive to osmotic shock, and were free of amino sugars and rigid cell walls. They were unable to regenerate cell walls and form colonies. They may be true protoplasts. Osmotic barriers in the "protoplasts" were preserved, and the "protoplasts" incorporated C(14)-l-alanine at about 60% of the rate of intact cells.

Alanine↗

PubMed1964

TOLERANCE OF STAPHYLOCOCCUS AUREUS TO SODIUM CHLORIDE.

Parfentjev, I. A. (Institute of Applied Biology, New York, N.Y.), and Anna R. Catelli. Tolerance of Staphylococcus aureus to sodium chloride. J. Bacteriol. 88:1-3. 1964.-The tolerance of Staphylococcus aureus to high concentrations of sodium chloride in liquid medium has been reported. We found that S. aureus grows at 37 C in Tryptose Phosphate Broth saturated with sodium chloride. No difference was noticed between possibly pathogenic and nonpathogenic strains. Under the conditions of our tests, no changes in the original properties of S. aureus strains occurred. In contrast, solutions of sodium chloride in distilled water were injurious to staphylococci and killed most of these organisms in 1 hr. Staphylococci were killed faster at 37 C than at room temperature in a solution of 0.85% sodium chloride in water. Addition of traces of Tryptose Phosphate Broth had a protective effect and prolonged the life of these organisms in physiological saline. All tests were performed at pH 7.2.

Pharmacology↗

PubMed1964

DIFFERENTIATION BY IMMUNODIFFUSION AND BY QUANTITATIVE IMMUNOFLUORESCENCE BETWEEN 5-FLUOROURACIL-TREATED AND NORMAL CELLS FROM A TOXINOGENIC STAPHYLOCOCCUS AUREUS STRAIN.

De Repentigny, J. (University of Montreal, Montreal, Quebec, Canada), S. Sonea, and A. Frappier. Differentiation by immunodiffusion and by quantitative immunofluorescence between 5-fluorouracil-treated and normal cells from a toxinogenic Staphylococcus aureus strain. J. Bacteriol. 88:444-448. 1964.-Immunodiffusion and quantitative immunofluorescence can both detect antigenic changes produced by 5-fluorouracil (FU) in Staphylococcus aureus Wood 46 strain. When FU is added to the cultures in their logarithmic phase of growth, a number of bacterial antigens are no longer detectable by immuno-diffusion and the intensity of the total immuno-fluorescence of bacteria is diminished; thus, these antigens are either profoundly modified or no longer synthesized. Uracil and, less effectively, thymine can reverse the FU inhibitory effect on the synthesis of antigens, and the number of precipitin lines remains closer to controls. The immunochemical approach provides a new way of obtaining information on the action of this pyrimidine analogue on metabolic processes in pathogenic bacteria. Microscopic quantitative immunofluorescence seems to be adaptable to give indirect information on changes in the metabolism or synthesis or antigens of a single bacterial cell.

Antigens↗

PubMed1964

VIABILITY AND METABOLISM OF STAPHYLOCOCCUS AUREUS AFTER FREEZING, LYOPHILIZATION, AND IRRADIATION.

Ahn, T. H. (University of California, Los Angeles), H. Nishihara, Charles M. Carpenter, and G. V. Taplin. Viability and metabolism of Staphylococcus aureus after freezing, lyophilization, and gamma irradiation. J. Bacteriol. 88:545-552. 1964.-Reproductive viability and oxidative activity on various substrates (alanine, glutamic acid, glucose, lactic acid) were studied in Staphylococcus aureus cells subjected to (i) freezing and thawing, (ii) lyophilization, (iii) irradiation in suspension, (iv) irradiation in lyophilized state, and (v) lyophilization and irradiation in the presence of various substrates. Freezing and thawing in dilute albumin solution had no effect on viable cell count and on lactic acid oxidation, but the oxidation rates of alanine, glutamic acid, and glucose were decreased. Lyophilization reduced the viable cell count to 25 to 30% of the initial value, and decreased the oxidation rates of glutamic acid and alanine proportionally, whereas decrease in glucose oxidation was less than proportional. The lactic acid oxidation rate was not affected by lyophilization. An irradiation dosage of about 40,000 r was required to reduce the viable count of cells irradiated in suspension by one log(10) unit; lyophilized cells required 140,000 to 170,000 r. Cells irradiated in suspension with dosages sufficient to render them completely nonviable on culture continued to respire on lactic acid at about 60% the rate of nonirradiated cells, but the residual activities on the other substrates tested were less than 10 to 12%. Lyophilized cells irradiated with sufficient dosages for cultural nonviability retained 40 to 80% of the oxidative capacity of nonirradiated cells on the test substrates. Cells lyophilized and irradiated in the presence of albumin generally retained a greater portion of their oxidative activities compared with cells lyophilized and irradiated in buffer. Cells irradiated in the presence of various oxidizable substrates gave varying results, depending both on the substrate present during irradiation and the substrate added for the oxidative studies.

Alanine↗

PubMed1964

INDUCIBLE RESISTANCE TO ERYTHROMYCIN IN STAPHYLOCOCCUS AUREUS.

Weaver, Judith R. (Iowa State University, Ames), and P. A. Pattee. Inducible resistance to erythromycin in Staphylococcus aureus. J. Bacteriol. 88:574-580. 1964.-The dissociated resistance of Staphylococcus aureus to erythromycin was examined and was found to possess the characteristics of an inducible enzyme. The induction of resistance to high concentrations of erythromycin in S. aureus occurred only after prior exposure to subinhibitory concentrations of erythromycin. The only macrolide antibiotic examined which induced resistance was erythromycin, and the resistance of induced populations was rapidly lost when they were grown in the absence of this antibiotic. Induction did not occur when protein synthesis was inhibited by either chloramphenicol or histidine starvation of a histidine auxotroph. The macrolide antibiotics inhibited the induction of resistance at the same minimal concentrations required to inhibit growth and induced synthesis of beta-galactosidase. Therefore, the mode of action of the macrolide antibiotics is to inhibit protein synthesis, and the induction of resistance overcomes this inhibition in some manner which is associated with the synthesis of new protein.

Anti-Bacterial Agents↗

PubMed1964

SENSITIVITY OF COCCAL AND L FORMS OF STAPHYLOCOCCUS AUREUS TO FIVE ANTIBIOTICS.

Kagan, B. M. (Cedars of Lebanon Hospital, Los Angeles, Calif.), Susan Zolla, R. Busser, and Silvija Liepnieks. Sensitivity of coccal and L forms of Staphylococcus aureus to five antibiotics. J. Bacteriol. 88:630-632. 1964.-Antibiotics whose primary site of action is in the cell wall (penicillin and cephalothin) do not inhibit growth of L-phase organisms. In this study, kanamycin, neomycin, polymyxin B, lincomycin, and gentamycin were found to be more active against L-phase growth of Staphylococcus aureus in vitro than against the coccal forms. Therefore, their primary site of antimicrobial activity appears to be other than that involved in the synthesis or integrity of the cell wall.

Anti-Bacterial Agents↗

PubMed1964

DENSITY-GRADIENT PATTERNS OF STAPHYLOCOCCUS AUREUS CELLS AND CELL WALLS DURING GROWTH AND MECHANICAL DISRUPTION.

Huff, Eskin (National Institute of Allergy and Infectious Diseases, Bethesda, Md.), Harriet Oxley, and Carol S. Silverman. Density-gradient patterns of Staphylococcus aureus cells and cell walls during growth and mechanical disruption. J. Bacteriol. 88:1155-1162. 1964.-Procedures capable of rapid disruption of Staphylococcus aureus cells with optimal release of intact cell walls were investigated. This search was implemented by observation of the flotation patterns of cells and subcellular particulate matter after centrifugation in a cesium chloride density gradient. A quantitative evaluation of the light-scatter throughtout the gradient was achieved by transfer of the entire density gradient into an optical cell with wedge-shaped cross section. When this cell was photographed under indirect illumination, each band of light-scattering material appeared on the negative as a shaded curve, with an area proportional to amount of that material present. A series of photographs of known amounts of cells and cell walls was used to estimate the amounts of these materials in mixtures of the two occurring during mechanical disruption. With the methods employed, time studies established the optimal time for release of cell walls as 5 min in a Braun shaker. The use of sucrose gradients in the further purification of cell walls, and chemical analysis of the isolated walls, are described.

Bacteriological Techniques↗

PubMed1964

LIPOLYTIC ACTIVITIES OF STAPHYLOCOCCUS AUREUS. I. NATURE OF THE ENZYME PRODUCING FREE FATTY ACIDS FROM PLASMA LIPIDS.

O'Leary, William M. (Cornell University Medical College, New York, N.Y.), and Julia T. Weld. Lipolytic activities of Staphylococcus aureus. I. Nature of the enzyme producing free fatty acids from plasma lipids. J. Bacteriol. 88:1356-1363. 1964.-The enzyme of Staphylococcus aureus, Guy strain, which is responsible for producing free fatty acids in human plasmas was purified approximately 225-fold from extracts of cultures in which the organism was grown. Purification procedures included ammonium sulfate precipitation, ethanol precipitation, gel filtration, and diethylaminoethyl-cellulose chromatography. Stability characteristics, pH and temperature optima, and substrate requirements were established. The principal plasma substrate is plasma lipoprotein, and the major reaction product is free octadecenoic acid. The ability of this compound to uncouple oxidative phosphorylation may be involved in the pathological effects of S. aureus at the molecular level.

Blood↗

PubMed1964

TYPES OF HEMOLYSINS PRODUCED BY STAPHYLOCOCCUS AUREUS, AS DETERMINED BY THE REPLICA PLATING TECHNIQUE.

Haque, Riaz-Ul (The Ohio State University, Columbus), and Jack N. Baldwin. Types of hemolysins produced by Staphylococcus aureus as determined by the replica plating technique. J. Bacteriol. 88:1442-1447. 1964.-The type of hemolysins produced by a representative number of cells of a culture of Staphylococcus aureus was determined by the replica plating technique, with the use of sheep, rabbit, human, and horse blood agar plates. Unless horse and human red blood cells were used, a distinction between alpha-and delta-hemolysins was difficult to make. Strains producing alpha-beta-, beta-delta-, and alpha-delta-hemolysins were recognized by the replica plating technique. A few colonies produced lysis of rabbit, horse, and human cells without lysis of sheep red blood cells. This pattern of lysis was not detected by the radial streak or the tube titration procedures when mixed populations were examined, and may be due to a lytic factor not hitherto described.

Animals↗

PubMed1965

PLASMA COAGULATION BY ORGANISMS OTHER THAN STAPHYLOCOCCUS AUREUS.

Bayliss, Berenice G. (Washington State University, Pullman), and Elizabeth R. Hall. Plasma coagulation by organisms other than Staphylococcus aureus. J. Bacteriol. 89:101-105. 1965.-Approximately 200 organisms were investigated for their ability to clot human and rabbit plasma. Various anticoagulants were used in preparing the plasma: acid-citrate-dextrose, ethylenediaminetetraacetate, balanced oxalate, potassium and sodium oxalates, and heparin. Twelve organisms were found which coagulated citrated plasma in less than 8 hr: four strains of Streptococcus faecalis; two strains of S. faecalis var. zymogenes; three strains of S. faecalis var. liquefaciens; and one strain each of S. pyogenes, Escherichia coli, and Serratia marcescens. Six strains of coagulase-positive Staphylococcus were selected for use as controls. Experiments were performed to determine the mechanism by which these microorganisms coagulated citrated plasma. As this was the only plasma clotted, it was presumed that the citrate was utilized by the microorganisms, thereby releasing the calcium which was then made available so that normal physiological clotting could occur. To test this hypothesis, a chromatographic method was employed to determine the presence or absence of citrate. Coagulation tests, by use of increasing amounts of citrate, showed a linear relationship between the amount of citrate in the plasma and the coagulation time. It was demonstrated that the organisms must be actively metabolizing to clot citrated plasma. Proof for this was obtained by using a cell-free filtrate, to which thimerosal had been added to inhibit growth, instead of whole cultures for the coagulation test. Only the coagulase-positive staphylococci coagulated the citrated plasma under these conditions. From the results obtained, it was concluded that plasma coagulation by these organisms was by citrate utilization.

Animals↗

PubMed1965

INCORPORATION OF HYDROXYLYSINE INTO THE CELL WALL AND A CELL-WALL PRECURSOR IN STAPHYLOCOCCUS AUREUS.

Smith, W. Grady (University of Minnesota, St. Paul), Daniel P. Gilboe, and L. M. Henderson. Incorporation of hydroxylysine into the cell wall and a cell-wall precursor in Staphylococcus aureus. J. Bacteriol. 89:136-140. 1965.-Recent work has shown that hydroxylysine can substitute for lysine in cell-wall synthesis of Streptococcus faecalis, apparently becoming incorporated into cell-wall mucopeptide. This paper extends these observations to investigate the metabolism of hydroxylysine in Staphylococcus aureus, an organism from which sufficiently large quantities of cell-wall precursors. uridine diphosphate-N-acetylmuramyl peptides, could be obtained. Hydroxylysine has been shown to be incorporated into the cell-wall precursor uridine diphosphate-N-acetylmuramyl l-ala.d-glu. l-lys.d-ala.d-ala from S. aureus (Copenhagen) apparently in lieu of lysine. Hydroxylysine was also incorporated into the cell-wall mucopeptides of S. aureus in resting cultures. This incorporation was inhibited by penicillin or lysine, but not by chloramphenicol. Hydroxylysine had little effect on the incorporation of lysine into S. aureus. Hydroxylysine acted as a growth inhibitor in this organism; the inhibition was reversed by lysine.

Alanine↗

PubMed1965

EFFECT OF HEMIN ON THE FORMATION OF THE CYTOCHROME SYSTEM OF ANAEROBICALLY GROWN STAPHYLOCOCCUS EPIDERMIDIS.

Jacobs, N. J. (Dartmouth Medical School, Hanover, N.H.), and S. F. Conti. Effect of hemin on the formation of the cytochrome system of anaerobically grown Staphylococcus epidermidis. J. Bacteriol. 89:675-679. 1965.-The cytochrome content and respiratory capacity of cells of a strain of Staphylococcus epidermidis grown in complex media under aerobic and anaerobic conditions were determined. Difference spectra revealed cytochromes of the a, b(1), and o types in aerobically grown cells. These cells respired at a rate of 66 muliters of O(2) per hr per mg of cells (dry weight). The cytochrome content of anaerobically grown cells was markedly reduced, and trace amounts only of cytochromes of the b(1) and o type were visible. The respiratory rate, with glucose as substrate, was reduced to about one-tenth of that observed in aerobically grown cells. Cells grown anaerobically in medium supplemented with hemin had as high a content of pigments resembling cytochromes b(1) and o as did aerobically grown cells; the respiratory rate was 60% of that observed in aerobically grown cells. Cytochrome of the a type, similar to that found in aerobically grown cells, was not detected.

Cytochromes↗