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Influence of the pH-value on the growth of Staphylococcus epidermidis, Staphylococcus aureus and Propionibacterium acnes in continuous culture.

A cutaneous isolate of Staphylococcus epidermidis, Staphylococcus aureus and Propionibacterium acnes was grown in continuous culture at varying pH-values ranging from 5.0 to 8.5. In terms of the specific growth rate as well as the bacterial density during the plateau phase there were remarkable differences. In particular, Propionibacterium acnes grew much better in the pH 6.0 to 7.0 range than in a more acidic or alkaline milieu. Staphylococcus epidermidis resembled Staphylococcus aureus showing no major difference at pH 5.5 and 7.0. These findings substantiate the hypothesis that minor changes of the pH in the pH 5.5 to pH 6.0 range as to be induced by chemically neutral or alkaline skin cleansers on the human skin surface can increase the number of propionibacteria but not staphylococci remarkably due to the relative alkalinity by itself.

Culture Media

In vitro adherence of Staphylococcus saprophyticus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus aureus to human ureter.

Staphylococcus saprophyticus adhered to human ureteral epithelium in vitro. The levels of adherence, which were determined quantitatively with the scanning electron microscope, correlated well with bacterial hemagglutinating activities with sheep erythrocytes (r = 0.9459, P < 0.01). Transmission electron microscopy revealed that the adhering bacteria and the hemagglutinating bacteria possessed similar pili-like structures on their cell surfaces. Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus aureus did not adhere to the epithelium. Only S. aureus adhered markedly to the connective tissue of the ureter, and adhesion of this organism was direct via its cell wall. This adherence test system clearly showed up differences in the abilities of these staphylococcal species to adhere to the urinary tract.

Bacterial Adhesion

Presence of an additional penicillin-binding protein in methicillin-resistant Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hominis, and Staphylococcus simulans with a low affinity for methicillin, cephalothin, and cefamandole.

The presence of an additional penicillin-binding protein (PBP) was demonstrated in methicillin-resistant strains of Staphylococcus epidermidis, S. haemolyticus, S. hominis, and S. simulans. In these four species, the apparent molecular mass of this protein was analogous to that of PBP 2' of methicillin-resistant S. aureus SR 1550-9. It exhibited a low affinity for methicillin, cephalothin, and cefamandole; and its synthesis was methicillin inducible. Peptide mapping of this PBP from the four species yielded identical results that were analogous to those obtained with S. aureus SR 1550-9. These results suggest that this protein is similar to, if not the same as, PBP 2' of S. aureus and that it is involved in methicillin resistance in the four species studied.

Bacterial Proteins

Analysis of the relationship between bacterial adherence and extracellular production of mannose, galactose, glucose and ribose in Staphylococcus epidermidis and Staphylococcus hominis.

Gas-liquid chromatography-mass spectrometry was used to analyze the extracellular extracts of 108 coagulase-negative staphylococcal strains for the presence of mannose, galactose, glucose and ribose, in order to determine whether production of these four monosaccharides, regarded as potential staphylococcal slime components, was associated with the adherence capacity of the individuals strains. A total of 90 Staphylococcus epidermidis and 18 Staphylococcus hominis strains were studied. Using the quantitative spectrophotometric assay, 21 Staphylococcus epidermidis strains were classified as strongly adherent, 12 as moderately adherent, 11 as weakly adherent, and 46 as nonadherent. All 18 Staphylococcus hominis strains were nonadherent. Mannose, galactose, glucose and ribose were detected as the main monosaccharide components in the extracellular extracts of all strains examined. Moreover, the mean relative concentrations of these monosaccharides were essentially the same for the different adherence phenotypes within the species Staphylococcus epidermidis. These results showed that there was no causal connection between the adherence of coagulase-negative staphylococci and the extracellular production of any of the four monosaccharides analyzed.

Bacterial Adhesion

Plasmid-mediated resistance to fosfomycin in Staphylococcus epidermidis.

Staphylococcus epidermidis strain BM2641, isolated from a patient, was resistant to penicillin G, methicillin, aminoglycosides, chloramphenicol, macrolide, lincosamide and streptogramin B-type (MLS) antibiotics, and to high levels of fosmycin. Resistance to forsfomycin and/or to MLS was lost at low frequencies either spontaneously or after curing with novobiocin. The plasmid DNA from BM2641 and its cured derivatives was purified, analyzed by agarose gel electrophoresis and transferred to a nitrocellulose sheet. Comparative analysis of the resistance phenotypes with the plasmid content of the strains indicated that fosfomycin and MLS resistance were encoded by plasmids pIP1842 (2.5 kb) and pIP1843 (2.6 kb), respectively. Southern hybridization with a probe specific for gene fosA of Serratia marcescens showed that the fosfomycin resistance determinant in Staphylococcus is not homologous to that of Gram-negative bacteria.

Conjugation, Genetic

Hemagglutination and adherence to plastic by Staphylococcus epidermidis.

Staphylococcus epidermidis is an important nosocomial pathogen responsible for intravenous catheter-related bacteremia and infections of other prosthetic medical devices. We found that the ability of S. epidermidis to hemagglutinate erythrocytes correlated with the adherence of bacteria to plastic and to intravenous catheters. S. epidermidis isolates responsible for prosthetic-valve endocarditis (n = 61) and isolates from intravenous catheters (n = 59) were significantly more likely to cause hemagglutination than isolates from the skin of preoperative cardiac surgery patients (n = 19) (P = 0.027). S. epidermidis isolates (n = 23) recovered from the skin of patients 7 to 10 days after cardiac surgery were significantly more likely to exhibit hemagglutination than the preoperative isolates (P = 0.015). By a quantitative adherence assay, we also observed that the hemagglutination titer and number of species of erythrocytes agglutinated correlated directly with adherence to polystyrene (P less than 0.001). In addition, hemagglutinating isolates were significantly more likely to be recovered in high number from intravenous catheters when semiquantitative catheter culture techniques were used (P less than 0.001). We speculate that hemagglutinin(s) either plays a direct role in adherence to polymers and thus prosthetic-device infection or serves as an easily demonstrable marker for adherence-prone isolates.

Animals

Methicillin-resistant Staphylococcus epidermidis.

Staphylococcus epidermidis is frequently associated with infection of prosthetic heart valves, prosthetic orthopedic devices, and neurosurgical shunts. Penicillinase-resistant semisynthetic penicillins, such as methicillin, have been the therapeutic and prophylactic agents of choice for S epidermidis infection. However, more S epidermidis isolates are now resistant to methicillin and other penicillins. In our laboratory 41% of S epidermidis isolates were resistant to methici-lin. All of the methicillin-susceptible isolates and 82% of the methicillin-resistant isoates were susceptible to cephalothin. Cephalothin should replace methicillin as the prophylactic and therapeutic agent of choice in institutions with a high percentage of methicillin-resistant S epidermidis.

Cephalothin

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.

Adhesiveness

Plasmids encoding for erythromycin ribosomal methylase of Staphylococcus epidermidis and Staphylococcus simulans.

Two 1.7 Md plasmids of Staphylococcus epidermidis and three ones of Staphylococcus simulans determining inducible macrolide-lincosamide resistance are identical as judged by restriction endonuclease fingerprinting. These plasmids designated pEI2101, pEI9105, pE1107, pEI1108 and pEI6104, respectively, belong to the incompatibility group 12. Dot-blot hybridization by photobiotin-labelled gene probe developed from S. aureus erythromycin ribosomal methylase gene showed cross hybridization between methylase-coding reference plasmids and the tested ones. The examined plasmids proved to be no transmissible in mating experiments into S. aureus recipients.

Drug Resistance, Microbial

Cloning and expression of methicillin resistance from Staphylococcus epidermidis in Staphylococcus carnosus.

A 6.2-kilobase chromosomal DNA fragment from a methicillin-resistant Staphylococcus epidermidis strain was cloned into Staphylococcus carnosus by using staphylococcal plasmid pCA44 as the vector. The recombinant plasmid obtained, pBBB21, conferred methicillin resistance on its host and was responsible for the synthesis of a low-affinity penicillin-binding protein (PBP), PBP 2'. PBP 2' determined by the S. epidermidis DNA and expressed as a membrane-bound PBP in S. carnosus reacted with monoclonal antibodies directed against PBP 2' of Staphylococcus aureus origin, and the cloned S. epidermidis DNA hybridized to the methicillin (mec)-specific DNA from S. aureus. These findings point to a common origin of the methicillin resistance determinant in staphylococci.

Bacterial Proteins

[Adhesion capacity and surface properties of Staphylococcus epidermidis and Staphylococcus saprophyticus].

We have comparatively studied the adherence of Staphylococcus epidermidis and S. saprophyticus to epithelial cells of a continuous cellular line of urinary origin (MDCK). The adherence of S. saprophyticus was significantly higher than that of S. epidermidis and this phenomenon was not related to the bacterial surface hydrophobicity or the ability of producing slime in vitro. The pre-incubation of the bacteria with sub-inhibitory concentrations of vancomycin, teicoplanin, amikacin, clindamycin, and roxitromycin did not modify the adherence ability of the microorganisms. Of the antimicrobial agents evaluated, only clindamycin significantly agents evaluated, only clindamycin, significantly inhibited the ability of producing slime of S. saprophyticus strains.

Animals

Staphylococcus epidermidis and Staphylococcus aureus in otitis media with effusion.

Bacteriologic investigation of middle ear effusion (MEE), external ear canal, and the nasopharynx was carried out on 458 patients with otitis media with effusion. Staphylococcus epidermidis was the most common bacteria in MEE, even after excluding the contaminants from the external ear canal, which had the same value of minimal inhibitory concentration as the paired MEE. The bacterial agreement of S epidermidis between MEE and the nasopharynx was extremely rare in contrast with Haemophilus influenzae, Streptococcus pneumoniae, and Branhamella catarrhalis, although the organism was also frequently isolated from the nasopharynx. Staphylococcus aureus, having the same minimal inhibitory concentration as that in the nasopharynx, was more frequently found in MEE than S epidermidis. The results suggest that S epidermidis found in MEE is not a pathogen, but rather a contaminant in many instances. Staphylococcus aureus seems to be a causative agent in otitis media with effusion.

Adolescent

[Nosocomial Staphylococcus epidermidis and Staphylococcus aureus septicemias in neonates].

Sixteen neonates developed staphylococcal septicemia (S. epidermidis in 10 cases and S. aureus in six). Two infections were due to maternofetal contamination and four to contaminated foreign material. Clinical symptoms included non-specific evidence of neonatal bacterial infection and, in S. aureus infections, suggestive skin or bone localizations. Fifteen patients recovered without sequelae and one died as a result of S. aureus septicopyemia. In view of the patterns of resistance to antimicrobial agents exhibited by S. aureus and S. epidermidis, the vancomycin-amikacin combination seems the most appropriate treatment in neonatal staphylococcal septicemias. However, the fosfomycin-cefotaxim combination can be proposed for the treatment of staphylococcal infections with osteoarticular or meningeal involvement.

Anti-Bacterial Agents

Deoxyribonucleic acid relatedness amongst Staphylococcus epidermidis and Staphylococcus saprophyticus strains.

The degree of binding was determined between DNA preparations from 65 staphylococci representing cluster defined in a numerical phenetic survey and 3H-labelled DNA from reference strains of S. epidermidis, S. haemolyticus, S. hominis, S. saprophyticus and S. warneri. The congruence between the DNA pairing and numerical phenetic data was good with S. epidermidis and S. saprophyticus being shown to be genomic species. However, some strains identified as S. epidermidis using recommended diagnostic procedures were found to belong to other taxa, notably S. simulans and S. warneri. The moles percent guanine plus cytosine content of the DNA of the test strains was within the range 27 to 34.

Base Composition

[Transfer of drug resistance plasmids from Staphylococcus epidermidis to Staphylococcus aureus in mixed culture].

A possibility of transfer of chloramphenicol-resistance and penicillin-resistance plasmids from 4 different donor S. epidermidis strains to 2 S. aureus strains was demonstrated. Chloramphenocol-resistance plasmid was found in S. epidermidis 1065/77 which was not expressed in this strain but could be transferred to and was expressed in Staphylococcus aureus strains. Penicillin-resistance and chloramphenicol-resistance plasmids were transferred simultaneously in 40 per cent of the colonies.

Chloramphenicol

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.

Animals

Adherence measured by microtiter assay as a virulence marker for Staphylococcus epidermidis infections.

Staphylococcus epidermidis strains isolated from clinical sources showed a wide range of abilities to adhere to glass and plastic materials. The degree of adherence depended on a number of factors, most notably, the composition of the growth medium. Adherence was enhanced by the addition of glucose or oleic acid to the growth medium and inhibited by serum. We have demonstrated a statistically significant association between the quantitative assessment of adherence to polystyrene tissue culture plates and clinical relevance. No such association was found when adherence was assessed by the qualitative adherence assay. Possible new approaches for assessing the clinical relevance of coagulase-negative staphylococcal isolates are discussed.

Bacterial Adhesion