Staphylococcus epidermidis infections

The opportunistic human pathogen Staphylococcus epidermidis has become the most important cause of nosocomial infections in recent years. Its pathogenicity is mainly due to the ability to form biofilms on indwelling medical devices. In a biofilm, S. epidermidis is protected against attacks from the immune system and against antibiotic treatment, making S. epidermidis infections difficult to eradicate.     

Phosphatase-negative Staphylococcus epidermidis

As a result of the study of a number of additional characteristics, more than a half of the cultures, formerly identified as phosphatase-negative S. epidermidis according to the scheme of the International Subcommittee on the Taxonomy of Staphylococci and Micrococci, have been shown to be more similar to S. Hominis and S. captis on account of their properties. The study of correlation between the main differentiating characteristics of S. epidermidis has shown that it is poorly pronounced. For this reason the decision to classify a phosphatase-negative culture with S. epidermidis should be based on the study of additional characteristics which differentiate this species not only from S. saprophyticus, but also from other phosphatase-negative staphylococci.     

Deoxyribonuclease-Positive Staphylococcus epidermidis Strains

Use of the agar plate test for the enzyme deoxyribonucleate 3′-nucleotidohydrolase (deoxyribonuclease) can result in frequent misdiagnosis of Staphylococcus epidermidis as S. aureus.     

Isolation of Staphylococcus epidermidis from Tobacco

Staphylococcus epidermidis has been isolated in significant numbers from growing tobacco leaves. The organism is also present on cured and aged tobacco.     

Mouse Virulent Strain of Staphylococcus epidermidis

Using 10⁹ or 10⁷ colony-forming units of a strain of Staphylococcus epidermidis (strain 1142) in saline or 5% mucin, respectively, 90 to 100% of mice died within 24 to 48 hr after intraperitoneal challenge infection. These organisms gradually multiplied in the peritoneal cavity when injected intraperitoneally into mice, while the mouse avirulent strain (strain 1124) rapidly decreased and no organisms were found there 20 hr after injection. This strain was capable of inducing resistance against challenge with homologous strains. The resistance appeared as early as the first week and disappeared the 4th week after the immunization. However, no resistance was induced with strain 1124 against challenge with strain 1142. Also, hyperimmune rabbit serum prepared with strain 1142 passively protected against challenge with homologous strain in mice. The protective antibody was absorbed out with homologous organisms but not with strain 1124. Subsequently, a surface substance was obtained from strains 1142 or 1124 by the method of Morse. The 1142 surface substance was capable of inducing a resistance against challenge with the homologous strain but not with the 1124 surface substance. Also, this substance absorbed the protective antibody in hyperimmune rabbit serum prepared with the homologous strain but not with the 1124 surface substance nor with the Smith surface antigen extracted from the Smith strain of Staphylococcus aureus. Conversely, the protective antibody in rabbit anti-Smith strain serum against challenge with the homologous strain was absorbed with the Smith surface antigen but not with the 1142 surface substance. In the agar diffusion test, the 1142 surface substance and the Smith surface antigen produced single precipitin lines only against homologous antisera. Biochemical analysis of the 1142 surface substance showed that the substance contained neither nucleic acids nor proteins but is composed of hexosamine, glycerol, phosphorus, alanine, glycine and phenylalanine.     

Adhesion of Staphylococcus epidermidis and Staphylococcus saprophyticus to a hydrophobic biomaterial

The relative surface charge and hydrophobicity of 16 strains of Staphylococcus epidermidis showed large variations. For this species no relationship between the two surface parameters was found. A highly negative surface charge was observed in all seven encapsulated strains (one S. epidermidis and six Staphylococcus saprophyticus strains). The adhesion of the staphylococci to fluorinated polyethylene-propylene films was not related to the relative surface charge and the hydrophobicity of the bacteria. On films pre-exposed to human plasma, the bacterial adhesion was substantially reduced. Mechanisms involved in the adhesion of coagulase-negative staphylococci to this biomaterial are discussed.     

MicroDSC study of Staphylococcus epidermidis growth

Background  

A microcalorimetric study was carried out using a Staphylococcus epidermidis population to determine the reproducibility of bacterial growth and the variability of the results within certain experimental parameters (temperature, bacterial concentration, sample thermal history). Reproducibility tests were performed as series of experiments within the same conditions using either freshly prepared populations or samples kept in cold storage. In both cases, the samples were obtained by serial dilution from a concentrated TSB bacterial inoculum incubated overnight.     

Anaerobic glucose and serine metabolism in Staphylococcus epidermidis

Anaerobically grown Staphylococcus epidermidis fermented glucose with the production of lactate and trace amounts of acetate, formate and CO2. Isotopic and inhibitor studies, assays for key enzymes of different metabolic pathways, and fermentation balances, all indicated that glucose was metabolized principally via glycolysis and to a very limited extent by the hexose monophosphate oxidative pathway. Serine fermentation proceeded via deamination and dismutation yielding NH3 and equimolar amounts of lactate, acetate and CO2; small amounts of formate arose by the operation of pyruvate-formate lyase. Incorporation of 0.5% (w/v) glucose in the growth medium depressed serine metabolism by repressing the activities of serine dehydratase and pyruvate dehydrogenase but, conversely, enhanced the activities of phosphofructokinase and lactate dehydrogenase. Glucose-grown organisms at various stages of anaerobic batch growth showed an inverse relationship between the rates of fermentation of serine and glucose. L-Lactate dehydrogenase activity in crude extracts depended on fructose 1,6-bisphosphate, and fructose 1,6-bisphosphate aldolase was found to be a class I aldolase. Despite the presence of ribokinase, D-ribose-5-phosphate isomerase, transaldolase and transketolase, the organisms utilized ribose only after growth aerobically in basal medium, and then at a slow rate after an initial lag period.     

Trimethoprim Resistance and Susceptibility Genes in Staphylococcus epidermidis

Genes encoding trimethoprim (TMP)-resistant and -susceptible dihydrofolate reductases (DHFR) in Staphylococcus epidermidis isolated in Saitama Prefecture were compared with the TMP-resistant DHFR gene of S. aureus, dfrA. The nucleotide sequences of TMP^r and TMP^S genes in five S. epidermidis isolates tested could be divided into three types: type 1, identical with the TMP^r gene dfrA that had been found in S. aureus; type 3, identical with the TMP^s gene dfrC in S. epidermidis; and type 2, having only two nucleotide substitutions to dfrC with no amino acid change. TMP^r isolates carried either one of the type 2 or type 3 sequences in addition to the type 1 sequence. A Southern hybridization analysis revealed that, in TMP^r S. epidermidis, the type 1 sequence was located on a 5.5 kb EcoRI-EcoRV restriction fragment together with the sequence for the gentamicin (GM)-resistant gene, while the type 2 or type 3 sequence was located on the 1.0 kb EcoRI-EcoRV fragment. No plasmid-carrying dfrA-homologous sequence was detected in the S. epidermidis isolates we tested. These results suggest that the TMP^r and GM^r genes are closely linked and located on the chromosome in S. epidermidis isolated in Japan.     

Isolation and Characterization of Bacteriophages Infecting Staphylococcus epidermidis

Bacteriophages infecting Staphylococcus epidermidis were isolated by mitomycin C induction. Three distinct phages (vB_SepiS-phiIPLA5, vB_SepiS-phiIPLA6, and vB_SepiS-phiIPLA7)—defined by plaque morphology, structure, virion proteins pattern, DNA restriction bands, and host range—were obtained. One-step growth curves of bacteriophages under optimal growth conditions for S. epidermidis F12 revealed eclipse and latent periods of 5–10 and 10–15 min, respectively, with burst sizes of about 5 to 30 PFU per infected cell. Transmission electron microscopy revealed that the phages were of similar size and belonged to the Siphoviridae family. Phage phi-IPLA7 had the broadest host range infecting 21 out of 65 S. epidermidis isolates. Phage phi-IPLA5 seemed to be a virulent phage probably derived from phi-IPLA6. Phages phi-IPLA5 and phi-IPLA7 exhibited increasing plaques surrounded by a halo that could be indicative of a polysaccharide depolymerase activity. Viable counts, determined during the infection of S. epidermidis F12, confirmed that phi-IPLA5 had a potent lytic capability and reduced S. epidermidis population by 5.67 log units in 8 h of incubation; in the presence of the mixture of phi-IPLA6 and phi-IPLA7, however, a reduction of 2.27 log units was detected     

Production and Purification of a Staphylococcus epidermidis Bacteriocin

Liquid cultures of Staphylococcus epidermidis 1580 contained rather small amounts of a bacteriocin, staphylococcin 1580, which was found both in the supernatant fluid and in the cell pellet. It could be extracted from the cells with 5% NaCl solution. The staphylococcin production could not be induced by ultraviolet irradiation or treatment with mitomycin C. Bacteria grown on semisolid medium produced a much larger amount of the compound with a high specific activity. The staphylococcin was purified by ammonium sulfate precipitation, ultracentrifugation, and chromatography on Sephadex columns. The purified material was homogeneous on polyacrylamide gel electrophoresis. The molecular weight was between 150,000 and 400,000. The bacteriocin was composed of protein, carbohydrate, and lipid and consisted of subunits exhibiting a molecular weight of about 20,000.     

Nature and Properties of a Staphylococcus epidermidis Bacteriocin

Staphylococcin 1580, produced by Staphylococcus epidermidis 1580, consisted of 41.8% protein, 34% carbohydrate, and 21.9% lipid. In the protein fraction, the acidic amino acids, glutamic and aspartic acid, and the neutral amino acids, glycine and alanine, predominated. Neutral sugars consisted of glucose, galactose, and fucose in a molar ratio of 6:3:1. The purified bacteriocin was not inactivated by heating for 15 min at 120 C in the presence of 0.5% serum albumin and was stable in the pH range from 3.5 to 8.5. The compound was sensitive to the action of the proteolytic enzymes trypsin, Pronase, and chymotrypsin. All gram-negative bacteria tested were resistant; a large number of gram-positive bacteria were sensitive to staphylococcin 1580 action. Growth of stable staphylococcal L-forms was inhibited by the bacteriocin to the same extent as their parent strains. The staphylococcin was adsorbed to cell walls, cell membranes, and resistant cells. The effect of staphylococcin 1580 appeared to be bactericidal but not bacteriolytic.     

Cloning and expression of Staphylococcus epidermidis urease gene sequences in Staphylococcus carnosus

The urease gene sequences of Staphylococcus epidermidis CNS23 were cloned and expressed in Staphylococcus carnosus TM300. In vitro translation of the cloned sequences revealed four polypeptides (60, 17, 11 and 7.5 kDa) that were associated with enzyme activity. Southern hybridisation experiments showed high homologies with the urease genes of Staphylococcus saprophyticus.     

Induction of resistance with heat-killed unencapsulated strains of Staphylococcus epidermidis against challenge with encapsulated strains of Staphylococcus epidermidis

Active immunization of mice with high doses of heat-killed unencapsulated strains of Staphylococcus epidermidis, which were grown in brain heart infusion media, protected mice against challenge with encapsulated strains of S. epidermidis. The unencapsulated strains were capable of absorbing the protective antibody in rabbit hyperimmune sera prepared with the encapsulated strains. Also, mice treated with rabbit hyperimmune sera prepared with the unencapsulated strains were protected against challenge with the encapsulated strains. The protective activities of these rabbit hyperimmune sera were assumed to be essentially identical to those of the protective antibody induced by the encapsulated strains.