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PubMed1999

Acute inflammation of the eyelid and cornea in Staphylococcus keratitis in the rabbit.

PURPOSE: The inflammatory response during Staphylococcus keratitis was analyzed biochemically and histologically to determine the source of the neutrophils infiltrating the tear film and cornea. METHODS: Rabbit eyes were swabbed and then examined by slit-lamp microscopy at 0, 5, 10, 15, 20, and 25 hours after intracorneal inoculation with Staphylococcus aureus. Bacterial colony-forming units were quantified in the cornea, eyelid, and acute inflammatory exudate. Myeloperoxidase activity of ocular swabs of acute inflammatory exudate, corneal homogenates, and eyelid homogenates was determined. Gross and microscopic examinations of corneas and eyelids were performed. RESULTS: The colony-forming units per cornea exceeded 10(7) after 10 hours, whereas no bacteria were cultured from the eyelid until 15 hours postinfection. Slit-lamp examination revealed progressive pathology, and the myeloperoxidase activities of ocular swabs, corneas, and eyelids increased markedly by 15 hours postinfection. Corneas showed a wave of neutrophils moving from the tear film toward bacteria in the central corneal stroma and early neutrophil migration from the limbus into the stroma. In the eyelid, neutrophils migrated from the stromal vessels to the tear film. CONCLUSIONS: Staphylococcus keratitis in the rabbit causes acute inflammation in the overlying eyelid. Neutrophils of the acute inflammatory exudate interact with the infected cornea, whereas neutrophils migrating through the cornea from the limbus remained distant from the site of infection.

Acute Disease↗

PubMed1963

INHIBITORY ACTION OF PHAGE K ON STAPHYLOCOCCAL DEHYDROGENASES. I. EFFECT ON VARIOUS STRAINS OF STAPHYLOCOCCUS AUREUS, INCLUDING MEMBERS OF THE PHAGE-TYPING SERIES.

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.

Bacteriophage Typing↗

PubMed1964

NATURE OF THE DETERMINANT CONTROLLING PENICILLINASE PRODUCTION IN STAPHYLOCOCCUS AUREUS.

Harmon, Shirley A. (Ohio State University, Columbus), and Jack N. Baldwin. Nature of the determinant controlling penicillinase production in Staphylococcus aureus. J. Bacteriol. 87:593-597. 1964.-From three penicillinase-producing strains of Staphylococcus aureus 38 penicillin-sensitive mutants were isolated. With phages 80 and 53 of the International Typing Series, each of these mutants was a competent recipient of the penicillinase marker from the parent strains. Although numerous reciprocal transductions were performed between penicillin-sensitive mutants, no penicillinase-producing recombinants were observed. Reversion of the penicillinase-negative mutants to capacity to produce penicillinase was never observed. When tested for 50 distinctive physiological characteristics, the mutants generally were similar to the parent strains except for resistance to penicillin. One penicillin-sensitive mutant failed to produce detectable amounts of five exoenzymes. By treating resistant strains with acridine orange, it was possible to isolate sensitive strains at a frequency that was 60 times greater than the spontaneous mutation rate. Thus, the determinant controlling the capacity of strains of S. aureus to produce penicillinase appears to be a cytoplasmic element.

Acridines↗

PubMed1964

NUTRITIONAL MUTATIONS AND TRANSDUCTION BY ULTRAVIOLET-INACTIVATED PHAGE IN STAPHYLOCOCCUS AUREUS.

Carere, A., and Isabella Spada-Sermonti (Istituto Superiore di Sanità, Rome, Italy). Nutritional mutations and transduction by ultraviolet-inactivated phage in Staphylococcus aureus. J. Bacteriol. 88:226-232. 1964.-By use of a semisynthetic medium, various nutritional mutants of Staphylococcus aureus 80 were isolated. Most of them require purine derivatives. From their mutation pattern, it may be inferred that guanine can be converted to both guanilic and adenilic acid, and adenine to the latter only. Guanosine, hypoxanthine, and inosine appear to be intermediate in the synthesis of adenilic acid from exogenous guanine. Transduction of several nutritional markers was obtained by streaking phage 80 obtained from the donor strain onto selective media heavily seeded with the recipient strain. To prevent lysis, the phage was previously ultraviolet-inactivated.

Adenine↗

PubMed1964

CELL-WALL LYSINS OF STAPHYLOCOCCUS AUREUS STRAINS INDUCED BY SPECIFIC TYPING PHAGES.

Ralston, Doris J. (University of California, Berkeley), and Mary McIvor. Cell-wall lysins of Staphylococcus aureus strains induced by specific typing phages. J. Bacteriol. 88:667-675. 1964.-At least 12 different phages induced the formation of soluble lysins (separable from the phages by ultracentrifugation). The lysins caused rapid clearing of heat-killed cells of all strains of Staphylococcus aureus tested, irrespective of the capacity of the phage to form plaques on living cells of the strain. The uninfected cells of the 12 strains contained a second lysin, an autolysin, released upon cellular autolysis. The autolysin preparations differed from the phage-induced lysins, in that they exhibited relatively high activity for lysing Micrococcus lysodeikticus and low activity for strain S. aureus K(1N), and were each specifically inhibited by antiserum prepared against purified autolysin from strain K(1). A third kind of lysin, virolysin, induced by polyvalent phage K, was differentiated from the lysins of the specific phages on the basis of its antigenic specificity and lack of action on M. lysodeikticus. All three kinds of lysins digested the mucopeptide portion of staphylococcal cell walls. No evidence was found that any of these lysins possessed specific host ranges which could be correlated with the lytic host range of the inducing phage.

Animals↗

PubMed1964

LYSIS-FROM-WITHOUT OF STAPHYLOCOCCUS AUREUS STRAINS BY COMBINATIONS OF SPECIFIC PHAGES AND PHAGE-INDUCED LYTIC ENZYMES.

Ralston, Doris J. (University of California, Berkeley) and Mary McIvor. Lysis-from-without of Staphylococcus aureus strains by combinations of specific phages and phage-induced lytic enzymes. J. Bacteriol. 88:676-681. 1964-Several typing phages, adsorbed in sufficient concentrations to their homologous propagating strains, altered the cell surface so as to render the cells sensitive to rapid and synergistic lysis by extra-cellular additions of wall lysins. Lysis was effected both by lysins induced by the individual phages and by phage K(1) virolysin. Phage K(1) also rendered cells sensitive to the lysins of the typing phages. With the exception of lysins from PS 53, 70, and 77, none of the lysins nor purified phages tested separately caused significant lysis of living cells. Lysis-from-without in Staphylococcus aureus appears to be a stepwise process: sensitization by phage followed by digestion of the wall by lysin.

Bacteriolysis↗

PubMed1965

SPECTROPHOTOMETRIC CHARACTERIZATION OF PIGMENTS PRODUCED BY STAPHYLOCOCCUS EPIDERMIDIS STRAINS ISOLATED FROM BOVINE UDDERS.

Sandvik, Olav (National Animal Disease Laboratory, Ames, Iowa), and Richard W. Brown. Spectrophotometric characterization of pigments produced by Staphylococcus epidermidis strains isolated from bovine udders. J. Bacteriol. 89:1201-1208. 1965.-Spectrophotometric analysis of methanol-extracted pigments of 70 isolates, representing 48 strains of Staphylococcus epidermidis, exhibited absorption curves that were classified into seven types, designated I, II, III, IV, V, VI, and a S. aureus type. Two subtypes were included in types I and III. All nonpigmented cell extracts were classified as type I. One of the six numbered type absorption curves was obtained with the majority of isolates. Three isolates gave an absorption curve that was similar to the curve produced by extracts of five of seven S. aureus strains, thus the designation S. aureus type. The differences in pigment complexes indicated by the various absorption curves of methanol extracts were substantiated by column-chromatography studies. Generally, pigments of types II to VI, as produced by representative strains, were of a xanthophyllic nature, whereas, the S. aureus type pigment studied was carotenelike. Analyses of representative strains showed that the type of spectral absorption curves did not change whether the organisms were carried in vitro and tested through 3 months, or isolated repeatedly from infected udders for periods up to 8 months. The method of determining the spectral absorption curves of whole-cell methanol extracts provides an additional tool for differentiating strains of S. epidermidis that can be used in enzootiological studies of udder infections.

Animals↗

PubMed1965

TRANSDUCTION OF STAPHYLOCOCCUS AUREUS TO TETRACYCLINE RESISTANCE IN VIVO.

Jarolmen, Howard (Hahnemann Medical College, Philadelphia, Pa.), Amedeo Bondi, and Richard L. Crowell. Transduction of Staphylococcus aureus to tetracycline resistance in vivo. J. Bacteriol. 89:1286-1290. 1965.-Staphylophage 80, propagated on a hospital strain of Staphylococcus aureus 80/81, has been shown to transduce antibiotic resistance markers to a variety of staphylococcal recipient strains in vitro. In an attempt to demonstrate transduction of penicillin and tetracycline resistance in mice, experiments were performed in which mice were injected intravenously with a pathogenic recipient strain, S. aureus N135, and subsequently with transducing phage by the same route. Periodic assays of organs from infected mice revealed that maximal bacterial concentrations were attained in kidneys 6 days after infection, at which time the transducing lysate, containing approximately 5 x 10(10) plaque-forming particles, was introduced. Isolation of tetracycline-resistant transductants from the kidneys of infected animals was facilitated by the therapeutic administration of tetracycline. In contrast, penicillin-resistant transductants, which produced penicillinase, were not found even when penicillin therapy was administered. Results showed that tetracycline-resistant transductants were recovered from as many as 40% of test animals in repeated experiments. Furthermore, in some of these mice the entire staphylococcal population of the kidneys was found to be tetracycline-resistant. Control infected animals which did not receive phage were uniformly negative for tetracycline-resistant staphylococci. The finding that phage levels were low or undetectable at a time when tetracycline-resistant organisms were recovered from test animals provided evidence that transduction had occurred in vivo.

Animals↗

PubMed1984

Increased energy expenditure by granulocytes during phagocytosis of Staphylococcus aureus compared with other staphylococci.

The heat produced by human neutrophils during phagocytosis of different strains of Staphylococcus was analyzed by a flow calorimetric technique. At the same bacteria/leukocyte ratio Staphylococcus aureus yielded heat effects about twice that of other saprophytic staphylococci tested. In parallel experiments the degree of phagocytosis of radiolabeled bacteria was evaluated: S. aureus strains were the least phagocytosed. These data indicate that, for a single phagocytosed bacterium, S. aureus strains yield heat effects three to four times greater than do the saprophytic staphylococci tested (P less than .001). Previous studies have shown that the heat produced by activated neutrophils derives essentially from the aerobic catabolism of glucose involved in the production of bactericidal O2 metabolites and is directly proportional to O2 consumption. The results reported here indicate that less efficient internalization of a staphylococcal strain is correlated with a greater energy expenditure for O2 metabolite formation during the metabolic burst.

Calorimetry↗

PubMed2000

Identification of a second lipase gene, gehD, in Staphylococcus epidermidis: comparison of sequence with those of other staphylococcal lipases.

The identification and molecular characterization of a previously unidentified lipase, gehD, from the human cutaneous commensal Staphylococcus epidermidis is reported. A lipase-GehC-deficient but otherwise isogenic mutant of S. epidermidis 9 was constructed by allele replacement. However, the mutant was found to retain 50% of the wild-type lipase activity in liquid culture. Rescreening of a genomic library revealed the presence of a second lipase gene, gehD, which was subsequently mapped and sequenced. In common with other staphylococcal lipases, GehD appeared to be translated as a 650-700 amino acid precursor which is processed post-translationally to an extracellular mature lipase of 360 amino acids with a size of approximately 45 kDa. Comparison of the amino acid sequence of GehD with those of other staphylococcal lipases revealed a high level of conservation between the mature lipase domains of different species. By hybridization studies, both gehC and gehD genes were found to be present in S. epidermidis isolates from both clinical and non-clinical backgrounds, but neither hybridized to DNA isolated from other staphylococcal strains. Construction of a phylogenetic tree and calculation of amino acid sequence homologies between mature lipases, however, suggested that the lipases of S. epidermidis may be more closely related to those of Staphylococcus aureus than to each other.

Amino Acid Sequence↗

PubMed2004

Novel non-mecA-containing staphylococcal chromosomal cassette composite island containing pbp4 and tagF genes in a commensal staphylococcal species: a possible reservoir for antibiotic resistance islands in Staphylococcus aureus.

Among methicillin-resistant Staphylococcus aureus isolates, a staphylococcal chromosomal cassette containing the mecA gene (SCCmec) is integrated into the chromosome at a unique site. SCCmec also contains unique ccrAB recombinase genes mediating its integration and excision from the genome and is flanked by characteristic left and right direct- and inverted-repeat sequences. A few non-mecA-containing SCC elements that have the other molecular features described above have recently been described. The origin of these cassettes is not clear. We have identified two new members of the SCC family integrated within orfX in Staphylococcus epidermidis strain ATCC 12228, neither of which carries mecA. One is a 57-kb element flanked by a unique 28-bp SCC direct repeat. It was called the SCC composite island (SCC-CI) because it carries a 19-kb SCC element (SCCpbp4) nested within it. SCCpbp4 contains pbp4 and tagF genes, as well as one pair of ccrAB genes (allotype 2) flanked by classical SCC-specific terminal repeats. External to SCCpbp4, SCC-CI contains a second pair of ccrAB genes (allotype 4), three IS431 elements, and genes mediating resistance to heavy metals. Genes mediating restriction-modification that may facilitate horizontal transfer are also present within SCC-CI, both within and outside SCCpbp4. Several novel arrangements of the SCC direct and inverted repeats were identified. Several long stretches of homology with other SCCs were found within and outside SCCpbp4. In view of the fact that SCC-CI was found in a commensal species, it may represent a reservoir for sequences involved in genetic shuffling between staphylococci and may contribute to the diversity found in SCC elements.

Amino Acid Sequence↗

PubMed1963

EFFECT OF SODIUM CHLORIDE CONCENTRATION IN AN AGAR MEDIUM ON GROWTH OF HEAT-SHOCKED STAPHYLOCOCCUS AUREUS.

The numbers of Staphylococcus aureus 196E surviving heat treatment in milk for various times at 60 C were determined by plate count with normal and modified Staphylococcus Medium No. 110 (S-110), with Plate Count Agar (PCA), and by milk enrichment techniques. Portions of specific heated samples appeared to contain lower populations of S. aureus 196E when the survivors were enumerated with normal S-110 than with PCA. Similarly, the times at 60 C needed for total destruction of the culture suspension in the test milk appeared to be shorter when survival was determined with normal S-110 agar. The apparently lower thermal death times were found to be related to the NaCl content of the S-110 medium, because use of S-110 agar containing lesser concentrations of NaCl resulted in growth of larger numbers of heat-shocked S. aureus 196E.

Agar↗

PubMed1983

Binding of fibronectin to Staphylococcus strains.

Fibronectin, a major protein component of plasma and loose connective tissue has previously been shown to bind to several strains of Staphylococcus aureus. We examined a large number of strains of different species of Staphylococcus with respect to their ability to bind fibronectin. The relative numbers of strains defined as fibronectin-binders among the different species were as follows: S. aureus (22 of 23), S. haemolyticus (5 of 5), S. warneri (8 of 11), S. hyicus (5 of 6), S. hominis (13 of 17), S. saprophyticus (11 of 20), S. epidermidis (4 of 7), and S. simulans (8 of 10). Only three species showed a predominance of nonbinders over binders: S. capitis (4 of 14), S. xylosus (0 of 4), and S. cohnii (3 of 11). These data indicate that staphylococcal species isolated from soft tissue infections frequently have the ability to bind fibronectin and suggest that the ability to bind to this protein may contribute to the virulence of coagulase-positive and coagulase-negative staphylococci.

Fibronectins↗

PubMed1961

Correlation between acid phosphatase and coagulase production or phage type of Staphylococcus aureus.

Pan, Yue-Liang (University of Michigan Medical School, Ann Arbor), and Harold J. Blumenthal. Correlation between acid phosphatase and coagulase production or phage type of Staphylococcus aureus. J. Bacteriol. 82:124-129. 1961.-A quantitative assay of acid phosphatase in 26 coagulase-negative and 73 coagulase-positive strains of Staphylococcus aureus was performed using an assay procedure modified to estimate extracellular, as well as the cellular, phosphatase. Some of the factors affecting the formation of acid phosphatase were studied. On the average, coagulase-negative strains produced 4.0 units and coagulase-positive strains 19.6 units although some negative strains produced more phosphatase than positive strains. More phosphatase was produced by strains of phage-group I than by strains of any other group.

Acid Phosphatase↗

PubMed1962

Transduction of resistance to some macrolide antibiotics in Staphylococcus aureus.

Pattee, P. A. (Iowa State University, Ames) and J. N. Baldwin. Transduction of resistance to some macrolide antibiotics in Staphylococcus aureus. J. Bacteriol. 84:1049-1055. 1962.-By use of phage 80 of the International Typing Series, propagated on appropriate strains of Staphylococcus aureus, two related markers controlling resistance to certain macrolide antibiotics (erythromycin, oleandomycin, spiramycin, and carbomycin) were transduced among a variety of strains of S. aureus. Unlike the markers controlling penicillinase production and resistance to chlortetracycline and novobiocin, the determinants of resistance to the macrolide antibiotics were transduced at normal frequencies (at least 300 transductants per 10(9) phage) only to certain of the recipient strains. One of the markers studied appears to control an inducible enzyme system which is specifically induced by sub-inhibitory concentrations of erythromycin and which controls resistance to erythromycin, oleandomycin, spiramycin, and carbomycin. The other marker examined confers resistance to erythromycin, oleandomycin, spiramycin, and carbomycin, and shows no evidence of being dependent upon an inducible mechanism.

Anti-Bacterial Agents↗

PubMed1962

Changes in immunodiffusion patterns and in nucleic acid content of Staphylococcus aureus grown in the presence of a nucleic acid fluorochrome.

Sonea, S. (University of Montreal, Montreal, Canada), J. de Repentigny, and A. Frappier. Changes in immunodiffusion patterns and in nucleic acid content of Staphylococcus aureus grown in the presence of a nucleic acid fluorochrome. J. Bacteriol. 84:1056-1060. 1962.-When grown in the presence of acridine orange, coagulase-positive Staphylococcus aureus exhibits changes in the number and the shape of its precipitin bands; these are more pronounced with commercial horse staphylococcal antitoxin than with normal human gamma-globulin. Simultaneously, there is an increase in the content of nucleic acids, especially deoxyribonucleic acid. Consequently, when grown in the presence of this nucleic acid fluorochrome, pathogenic staphylococci become similar to nonpathogenic strains in their antigenic structure and nucleic acid content. The bacterial population becomes a mixture of red and green fluorescing cells of different sizes. After centrifugation, most of the large and red fluorescing cells are found in the top layer, which shows more changes in antigenic composition and in nucleic acid content than the bottom layer. In subcultures grown in the absence of acridine orange, the cells revert to the original composition of the pathogenic strains.

Acridines↗

PubMed1963

Identity of Staphylococcus epidermidis.

Jones, Dorothy (American Meat Institute Foundation, Chicago, Ill.), R. H. Deibel, and C. F. Niven, Jr. Identity of Staphylococcus epidermidis. J. Bacteriol. 85:62-67. 1963.-The species Staphylococcus epidermidis is defined more precisely, and compared with other staphylococci. The physiological characteristics considered to be of primary import are: (i) ability to grow anaerobically in a defined complex medium with glucose or pyruvate as the energy source; (ii) inability to produce coagulase; (iii) ability to ferment serine as an energy source; (iv) the requirement for biotin and uracil (under anaerobic conditions) in a semisynthetic medium; and (v) inability to reduce nitrate beyond nitrite.

Coagulase↗

PubMed1963

CHARACTERISTICS OF A STRAIN OF STAPHYLOCOCCUS AUREUS GROWN IN VIVO AND IN VITRO.

Beining, Paul R. (The Catholic University of America, Washington, D.C.) and E. R. Kennedy. Characteristics of a strain of Staphylococcus aureus grown in vivo and in vitro. J. Bacteriol. 85:732-741. 1963.-A comparative survey was conducted on the characteristics of a strain of Staphylococcus aureus after it had been grown in vitro (VSB) and after it had been collected from the peritoneal exudate of an infected guinea pig (GSB). Both VSB and GSB strains gave the same results when studied in an extensive series of tests, including bound and soluble coagulases, bacteriophage type, antibiotic-sensitivity pattern, the usual fermentation reactions, deoxyribonucleic acid base composition, and qualitative tests for hemolysins, deoxyribonuclease, ribonuclease, staphylokinase, staphyloprotease, lipase, and phosphatase. The in vivo strain differed significantly from the in vitro strain in respiratory rate, agar gel diffusion studies, agglutinability in tube tests, virulence tests in rabbits and mice, growth on tellurite-glycine agar, susceptibility to human gamma-globulin in agar, and in the quantitative production of deoxyribonuclease, alpha-hemolysin, leucocidin, and hyaluronidase.

Agar↗