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Recurrent Staphylococcus aureus chalazia in hyperimmunoglobulinemia E (Job's) syndrome.

PURPOSE: To report a case of recurrent Staphylococcus aureus chalazia in a patient with hyperimmunoglobulinemia E syndrome (Job's Syndrome). DESIGN: Case report. METHODS: Three separate surgical incisions and curettages of multiple, recurrent chalazia of the right upper eyelid were performed over a course of 3 months. Cultures and pathologic specimens were obtained. Postoperative treatment consisted of oral erythromycin and amoxicillin/clavulonate, topical tobramycin/dexamethasone ointment, and warm compresses. RESULTS: Pathology of the tarsus confirmed the diagnosis of multiple chalazia. Cultures of the chalazia contents were positive for Staphylococcus aureus. No further recurrence was observed following the third surgical procedure over a 3-month follow-up period. CONCLUSION: Characteristic Staphylococcus aureus skin infections in immunodeficient patients with hyperimmunoglobulinemia E syndrome can involve the eyelids and may be recurrent despite appropriate medical and surgical therapy.

Adolescent↗

Biochemical and molecular characterization of Staphylococcus xylosus lipase.

The Staphylococcus xylosus strain secretes a non-induced lipase in culture medium: S. xylosus lipase (SXL). Pure SXL is a monomeric protein (43 kDa). The 23 N-terminal amino acid residues were sequenced. This sequence is identical to that of Staphylococcus simulans lipase (SSL); in addition, it exhibits a high degree of homology with Staphylococcus aureus lipase (SAL NCTC 8530) sequences. The cloning and sequencing of gene part encoding the mature lipase shows one nucleotide difference with SSL, which corresponds to the change of one residue at a position 311. The lipase activity is maximal at pH 8.2 and 45 degrees C. SXL is able to hydrolyse triacylglycerols without chain length specificity. The specific activity of about 1900 U/mg was measured using tributyrin or triolein as substrate at pH 8.2 and at 45 degrees C in the presence of 2 mM CaCl2. In contrast to some previously characterized staphylococcal lipases, Ca2+ is not required to trigger the activity of SXL. SXL was found to be stable between pH 5 and pH 8.5. The enzyme maintains 50% of its activity after a 15-min incubation at 60 degrees C. Using tripropionin or vinyl esters as substrates, SXL does not present the interfacial activation phenomenon. Unlike many lipases, SXL is able to hydrolyse its substrate in the presence of bile salts or amphiphilic proteins. SXL is a serine enzyme, which is inhibited by THL.

Amino Acid Sequence↗

Superiority of surgical versus medical treatment in patients with Staphylococcus aureus infective endocarditis.

BACKGROUND: We present here the clinical features and outcome of 54 patients affected by a Staphylococcus aureus infective endocarditis at the Amiens hospital between 1990 and 2000. The patients operated-on, group A (20 patients), were compared to the population of patients treated by exclusive antibiotherapy, group B (34 patients). PATIENTS AND METHOD: The male gender predominated with a sex ratio of 2.6. The mean age of the global population was 58.7+/-1.6 years. Time between onset of endocarditis symptoms and treatment (entire group) ranged from 1 to 120 days (mean 14.4 days). The main portal of entry were, respectively, for group A and group B: cutaneous 55% and 44.1%; intravascular material 5% and 8.8%; and rhinopharynx 5% and 8.8%. Seventy-five percent of the Staphylococcus aureus isolated were Methi-S. The main surgical treatment indication were: hemodynamic failure (HF) (30%), unstable infection with collapse (UI) (30%), UI+HF (10%), voluminous vegetation (20%) and embolism event (10%). RESULTS: The hospital mortality rate were respectively for the entire group, group A and group B: 25%, 35% and to 41% (ns). For group A, the operative mortality was lower(21%) after the first week. The actuarial survival rate (Kaplan-Meier) after 24 months was 54./+/-6.9% for the global population and 74+/-10.6% for group A and 43+/-8.5 for group B (p<0.001). The multivariate analysis finds severe sepsis and index of comorbidity as independent factors related to the global late mortality and, respectively, the age and the severe sepsis for group A, and the cardiac insufficiency for group B. CONCLUSION: The surgical treatment seems to be the best way to improve the results after Staphylococcus aureus endocarditis. The severity of the sepsis remains the most severe prognostic element, whatever the treatment adopted may be.

Anti-Bacterial Agents↗

[Antibiotics resistance of meticilline-resistant Staphylococcus aureus: detection of the first glycopeptides low sensibility strains in Tunisia].

The adaptation of Staphylococcus aureus to the hospital environment led to the acquisition of resistance to all antibiotics available in clinical practice. The aim of this study was to investigate the methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in the F. Bourguiba hospital of Monastir (Tunisia). We determined the antibiotype of all Staphylococcus aureus strains identified. Susceptibility rates to fosfomycin, chloramphenicol, rifampicin and pristinamycin were 7%, 3%, 2% and 0%, respectively. The prevalence of MRSA was 15.5% (96 strains); their susceptible to gentamicin progressively increased. The minimum inhibitory concentration (MICs) of oxacillin, vancomycin and teicoplanin were evaluated for the 96 MRSA strains. We identified two MRSA strains (M4 and M41) showing reduced glycopeptides susceptibility. Further analysis revealed that M4 and M41 harbor the gene encoding the class S and class F proteins specific for the Panton-Valentine Leukocidin (PVL). The mecA gene was detected only in strain M41 which harbors the Staphylococcal Cassette Chromosome (SCCmec) type III. This is the first reported MRSA showing reduced susceptibility to glycopeptides in Tunisia. Regulatory surveillance of susceptibility to antibiotics is needed to reduce the morbidity and the mortality rates as well as societal costs of S. aureus infections.

Acclimatization↗

Description of complete DNA sequence of two plasmids from the nukacin ISK-1 producer, Staphylococcus warneri ISK-1.

We report the whole DNA sequence of two plasmids, pPI-1 (30.2 kb) and pPI-2 (2.8 kb). These plasmids are from Staphylococcus warneri ISK-1, which produces a lantibiotic, nukacin ISK-1. Curing of pPI-1 resulted in a loss of bactericidal activity in the culture supernatant and the host's immunity to nukacin ISK-1, suggesting that the biosynthetic genes of the bacteriocin are encoded by pPI-1. Based on the results of a homology search of each open reading flame, pPI-1 is comprised of the following four distinct regions: (1) the nukacin ISK-1 biosynthesis and immunity gene cluster, (2) the thioredoxin gene cluster, (3) the replication region, and (4) a region of Staphylococcus epidermidis ATCC 12228, highly homologous to pSE-12228-05. Gene organization in the nukacin ISK-1 biosynthesis and immunity gene cluster is different from that in other lacticin-481 type gene clusters. The features of the replication protein encoded in the replicating region are somewhat different from other staphylococcus theta-replicating plasmids. pPI-2 comprised a disinfectant resistant gene, qacC, and the whole DNA sequence showed significant similarity to those of other qacC plasmids such as pSK108, suggesting that pPI-2 belongs to the qacC plasmid group.

Amino Acid Sequence↗

The smr gene resides on a novel plasmid pSP187 identified in a Staphylococcus pasteuri isolate recovered from unpasteurized milk.

UNLABELLED: This work describes a novel plasmid pSP187 (5550 bp) carrying the small multidrug resistance determinant smr encoding resistance to quaternary ammonium compounds (QACs). pSP187 was identified in a Staphylococcus pasteuri isolate recovered from bulk milk in a dairy cattle herd in Norway. Sequence analysis revealed 6 putative ORFs in addition to the smr gene within a cassette with identical genetic organization to that found in the pSK41-like Staphylococcus aureus plasmid pTZ22. A protein homology search suggested the gene product of ORF7 to be a putative replication initiation protein, while ORF2 was predicted to encode a protein homologous to members of FtsK/SpoIIIE cell division-DNA segregation protein families. Sequence similarities to some initiator proteins of rolling circle replicons (RCR) indicated that pSP187 uses a RCR mode of replication, supported by the detection of intermediate ssDNA using S1 nuclease treatment and hybridization analysis. Interestingly, a 30-bp sequence found upstream from ORF7 showed high similarity to other dyad symmetry motifs proposed as putative double-strand origins of replication in the plasmids pGI3 (Bacillus thuringiensis), pSTK1 (Bacillus stearothermophilus), and pER1-2 (Streptococcus thermophilus). IN CONCLUSION: The novel smr-containing plasmid pSP187 is the first member of RCR group VI to be identified in a Staphylococcus sp.

Amino Acid Motifs↗

Protamine sulfate and vancomycin are synergistic against Staphylococcus epidermidis prosthesis infection in vivo.

We have previously demonstrated that the quaternary amine, protamine sulfate (PS), is bactericidal against Staphylococcus epidermidis. In an attempt to decrease genitourinary prosthesis infection rates, we examined the ability of PS as a wound irrigant to inhibit Staphylococcus epidermidis viability. Eighty-seven Sprague-Dawley rats were studied by implanting a sterile silicone pellet in their dorsum. The pellet was inoculated with Staphylococcus epidermidis and the rats were divided into four groups based on the wound irrigant employed after inoculation: (1) control (sterile water) (2) vancomycin; (3) PS; (4) vancomycin + PS. All rats received perioperative and daily intramuscular vancomycin, and the pellets were explanted on postoperative day 28 and cultured. The infection rates were: (1) control 77%, (2) vancomycin 50%, (3) protamine sulfate 67%, and (4) protamine sulfate and vancomycin 19%. The differences between (2) vancomycin versus (4) vancomycin + PS and (3) PS versus (4) vancomycin + PS were significant (p = 0.05 and p < 0.005). The data suggest that PS potentiates vancomycin as a wound irrigant in prosthesis implantation.

Animals↗

The clinical significance of Staphylococcus aureus bacteriuria.

To investigate the rate of occurrence, clinical presentation, predisposing factors and frequency of secondary bacteremia 132 patients with significant Staphylococcus aureus bacteriuria were reviewed retrospectively. Staphylococcus aureus accounted for 3.3 per cent of all positive urine cultures. Most patients were elderly men. The most important predisposing factors in the urinary tract were indwelling catheters (63 per cent), obstruction (56 per cent) and instrumentation or surgery (43 per cent). Bacteremia developed secondary to bacteriuria in all 11 patients (8.3 per cent). For that reason Staphylococcus aureus bacteriuria should be regarded as a hazardous condition, especially in patients with predisposing factors in the urinary tract.

Adult↗

Staphylococcus saprophyticus: role in urinary tract infections in men.

We investigated the role of Staphylococcus saprophyticus in urinary tract infections in men. Of 9,314 urine specimens cultured from men in a veterans hospital only 3 (2 patients) yielded Staphylococcus saprophyticus. Thus, Staphylococcus saprophyticus rarely causes infection in older men and should not be sought routinely by microbiology laboratories in hospitals serving men primarily.

Humans↗

Molecular cloning and expression of a novel Staphylococcus aureus antigen.

A novel gene encoding an antigen from Staphylococcus aureus was isolated from an expression library by screening with antisera from patients with deep Staphylococcus aureus infections. In one positive clone an open reading frame, named ORF-2, was identified. Recombinant ORF-2 protein reacted with human immune serum. ORF-2 was shown to be present in other Staphylococcus aureus strains, but not in related species.

Amino Acid Sequence↗

Effect of proteolytic starter cultures of Staphylococcus spp. on biogenic amine formation during the ripening of dry fermented sausages.

The effect of proteolytic starter cultures of Staphylococcus carnosus and Staphylococcus xylosus on biogenic amine production was examined during the fermentation process of dry sausages. Microbial counts (lactic acid bacteria, Micrococcaceae and Enterobactenaceae), pH, moisture and proteolysis-related parameters were also studied. The polyamines spermine and spermidine were the main amines found in the raw material and they only showed slight fluctuations during the fermentation. The four elaborated batches presented a significant (P < 0.001) formation of tyramine and putrescine. The main rate of amine production was during the first three days, when a sharp pH decrease and the development of lactic acid bacteria occurred. Sausages fermented with starters had lower amounts of tyramine than naturally fermented sausages (control), but differences in the Micrococcaceae counts were only significant during the first week of the ripening process. A slight formation of diaminopropane, cadaverine, agmatine, tryptamine and phenylethylamine was observed. The amounts of histamine were constant and remained below 0.5 mg/kg of dry matter, while serotonin, octopamine and dopamine were not detected. The sausages with Staphylococcus as starter culture showed strong proteolysis that was correlated with higher pH values than those of the control sausages. However, no positive correlation was found between the proteolysis index and biogenic amine production. Since proteolysis was stronger during the second half of the ripening process, the release of free amino acids as amine precursors occurred later than the early amine production.

Animals↗

Curing of plasmid pE194 with novobiocin and coumermycin A1 in Bacillus subtilis and Staphylococcus aureus.

Plasmid pE194 determining macrolide-lincosamid-streptogramin B resistance could be eliminated with novobiocin and coumermycin A1 in Bacillus subtilis and Staphylococcus aureus. In both organisms the curing effect depended on the temperature. In the case of Staphylococcus aureus it increased very much at high temperatures, while in the case of Bacillus subtilis only a moderate increase could be observed. The curing activity of coumermycin was substantially weaker than that of novobiocin in both bacteria. This difference decreased at higher temperatures only in Staphylococcus aureus. In Bacillus subtilis the curing with novobiocin could be antagonized with coumermycin in accordance with the growth experiments reported previously.

Aminocoumarins↗

Limited effectiveness of chlorhexidine based hand disinfectants against methicillin-resistant Staphylococcus aureus (MRSA).

Hand disinfectants containing chlorhexidine are thought to be less bactericidal against methicillin-resistant Staphylococcus aureus (MRSA) than methicillin-susceptible Staphylococcus aureus (MSSA). We report an in vitro comparison between three distinct MRSA strains and three MSSA strains. The bactericidal efficacy of chlorhexidine digluconate, 'Hibiscrub' and 'Hibisol' against Staphylococcus aureus was determined in a quantitative suspension test. Logarithmic reduction factors (RF) were calculated for each of six parallel experiments. Chlorhexidine digluconate and 'Hibisol' showed RF > 5 at most concentrations and reaction times but 'Hibiscrub' did not. MRSA was found to be significantly less susceptible than MSSA to chlorhexidine digluconate, 'Hibiscrub' and 'Hibisol' (P < 0.05; two-tailed t-test for independent samples). 'Hibisol' was significantly more effective against MRSA than 'Hibiscrub' (P < 0.05). Hand disinfectants containing both alcohol and chlorhexidine (e.g., 'Hibisol') are more effective against MRSA than scrubs based only on chlorhexidine ('Hibiscrub') and should be used in clinical practice.

Chlorhexidine↗

[Epidemiology of Staphylococcus aureus nosocomial infections in a high-risk neonatal unit].

Nosocomial infections are a significant cause of morbidity and mortality throughout the world. In developing countries it is difficult to carry out effective surveillance and control programs for this type of infection because of the cost in both human and material resources. These considerations prompted us to perform a prospective study to determine the epidemiologic and microbiologic characteristics of nosocomial infections due to Staphylococcus aureus in the High-risk Neonatal Unit (HRNU) of the Instituto Autónomo Hospital Universitario de Los Andes (IAHULA), during the period of November 1997 to October 1998. Among a total of 120 microorganisms, 24 (20%) strains of Staphylococcus aureus were isolated; 47% were recovered from blood and 33% from conjunctive samples. Among the cases of conjunctivitis, S. aureus was the only pathogen isolated in 42%. Twenty of the 24 Staphylococcus aureus strains (83%) were methicillin-resistant (MRSA). According to their resistance profiles, we established 12 groups of strains from neonates with nosocomial infections and 1 group of strains from the two carriers among the healthcare personnel detected by microbiological screening. The MeRGmR pattern was the most frequent. Plasmid analysis disclosed two profiles, each having a plasmid molecular weight over 23.130 bp. The MRSA strains isolated from the neonates and those isolated from the carriers showed the same plasmid profile. This suggests that the healthcare personnel may have acted as reservoirs of the MRSA strains found in neonates with nosocomial infection.

Cross Infection↗

Characterization of RAP, a quorum sensing activator of Staphylococcus aureus.

Staphylococcus aureus are Gram-positive bacteria and cause diverse serious diseases in humans and animals through the production of toxins. The production of toxins is regulated by quorum sensing mechanisms, where proteins such as RNAIII activating protein (RAP) are secreted by the bacteria and induce virulence. Antibodies to RAP have been shown to protect mice from infection, but the molecular structure of RAP was not known and hindered vaccine development. To characterize RAP, recombinant protein was made and tested for its ability to induce genes important for pathogenesis (agr). In addition, monoclonal antibodies were produced to identify its cellular localization. Results shown here indicate that RAP is a 277-aa protein that is an ortholog of the ribosomal protein L2. Like the native molecule, recombinant RAP activates the production of RNAIII (encoded by agr). Using RAP specific monoclonal antibodies we demonstrate that RAP is continuously secreted and while RAP is expressed also in other bacteria (like Staphylococcus epidermidis, Staphylococcus xylosus and Escherichia coli), it is secreted to the culture medium only by S. aureus. Our results show that the ribosomal protein L2 has an extraribosomal function and that when secreted RAP acts as an autoinducer of virulence to regulate S. aureus pathogenesis.

Adaptor Proteins, Signal Transducing↗

Interaction between biofilms formed by Staphylococcus epidermidis and quinolones.

The interaction between pefloxacin, ciprofloxacin, norfloxacin, and ofloxacin and biofilms formed by Staphylococcus epidermidis (20 clinical isolates) was studied. In the presence of 1/2-MIC and 1/8-MIC of quinolones, the optical density of the biofilms was reduced to 22-24% and 65-74% of the controls, respectively. Treatment of preformed biofilms with quinolones in concentrations ranging from 12.5 microg/ml to 400 microg/mL caused reduction in the optical density of the adherent biofilms to 45-77% of the control. In an in vitro model of vascular catheter colonization, subinhibitory concentrations (12, 14 and 1/8 MIC) of fluoroquinolones reduced the number of adherent bacteria to 24-28%, 48-55% and 58-76% of the controls, respectively. The vascular catheter segments precolonized with Staphylococcus epidermidis for 24 h and exposed to the fluoroquinolones in 8-16 times MIC (100 microg/mL) for 2 h showed no growth of adherent cells. The activity of pefloxacin in reducing the bacterial adhesion and eradicating the preformed biofilms was demonstrated by scanning electron microscope. These data show that subinhibitory concentrations of ciprofloxacin, norfloxacin, pefloxacin, and ofloxacin inhibit the adhesion of Staphylococcus epidermidis to plastic surfaces and vascular catheters. Higher concentrations of fluoroquinolones were able to eradicate the preformed biofilms on vascular catheters.

Anti-Infective Agents↗

Mupirocin resistance in methicillin-resistant Staphylococcus aureus clinical isolates in a Spanish hospital. Co-application of multiplex PCR assay and conventional microbiology methods.

A total of 1785 Staphylococcus aureus clinical isolates were collected in our hospital during 1998 (526), 1999 (564) and 2000 (695). Among them, one hundred and thirty (39, 33 and 58, respectively) were phenotypically assigned as methicillin-resistant Staphylococcus aureus (MRSA); sixteen of these isolates (3, 2 and 11, respectively) were detected as highly mupirocin-methicillin resistant Staphylococcus aureus (MMRSA). In this work, our goal was to characterize MRSA and MMRSA clinical isolates by co-application of phenotypic and genotypic methods in order to determine the MMRSA incidence in our hospital during the period 1998-2000. With this purpose we compared and integrated the results obtained using conventional microbiology methods with those obtained using a multiplex polymerase chain reaction (PCR) assay. Our results showed a good complementation between these two approximations to determine the incidence of MMRSA clinical isolates and permitted to estimate that such incidence increased from 7.7% in 1998 to 19% in 2000.

Bacterial Proteins↗

In vitro and in vivo study of fosfomycin in methicillin-resistant Staphylococcus aureus septicaemia.

Five hundred strains of methicillin-resistant Staphylococcus aureus were tested against various anti-staphylococcal agents. Vancomycin, fusidic acid and fosfomycin were found to be the most effective. Only 1 strain out of 500 was resistant to fosfomycin. Three patients with methicillin-resistant Staphylococcus aureus septicaemia were successfully treated by fosfomycin. We conclude that fosfomycin could be the drug of choice for methicillin-resistant Staphylococcus aureus infection.

Adult↗