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PubMed2001

Secretion of human growth hormone by the food-grade bacterium Staphylococcus carnosus requires a propeptide irrespective of the signal peptide used.

Staphylococcal exoproteins can be divided into two groups. One group comprises proteins bearing only a signal peptide, the other group requires an additional propeptide for secretion. The secretion signals of the propeptide-requiring lipase from Staphylococcus hyicus (Lip) have been frequently used to produce recombinant secretory proteins in the food-grade species Staphylococcus carnosus. However, it has been unclear whether recombinant proteins can be secreted using signal peptides of staphylococcal proteins without propeptide. The human growth hormone protein (hGH) was fused to various staphylococcal secretion signals of proteins without propeptide (Seb, SceA, and SceB) and of proteins requiring a propeptide (lipase, lysostaphin, and glycerol ester hydrolase). Secretory hGH was efficiently produced by S. carnosus after fusion with any propeptide-containing secretion signal, whereas precursor proteins were retained in the cells when only a signal peptide was used. Addition of the first six amino acid residues of mature SceA to the signal peptide did also not lead to secretion of hGH. It was concluded that the properties of the mature protein domains determine whether a propeptide is required for secretion or not. The Lip propeptide could be efficiently removed from hGH after introduction of an enterokinase cleavage site between the two protein domains.

Amino Acid Sequence↗

PubMed2006

A newly isolated organic solvent tolerant Staphylococcus saprophyticus M36 produced organic solvent-stable lipase.

Thirty-eight high lipase activity strains were isolated from soil, seawater, and Brassica napus. Among them, a novel organic solvent tolerant bacterium (strain M36) was isolated from the seawater in Jiangsu, China. Isolate M36 was able to grow at high concentration of benzene or toluene up to 40% (vol/vol), and later identified as Staphylococcus saprophyticus by biochemical test and 16s ribosomal DNA sequence. No work on Staphylococcus producing lipase with organic solvent tolerance has been reported so far. The lipase of strain M36 whose activity in liquid medium was 42 U mL(-1) at 24-h incubation time was stable in the presence of 25% (vol/vol) p-xylene, benzene, toluene, and hexane.

Benzene↗

PubMed1996

Effect of Staphylococcus epidermidis on hydrogel contact lens retention on the rabbit eye.

A slime-producing isolate of Staphylococcus epidermidis attached to FDA Group II hydrogel contact lenses persisted on rabbit eyes for up to 14 days, but except for minor redness of the eye no other effect was observed. Eye flora of eight representative New Zealand White rabbits included four different species of Staphylococcus including S. epidermidis and one species of Micrococcus, none of which produced overtly obvious biofilms. The slime-producing strain of S. epidermidis adhered more effectively to lenses than a non-slime-producing strain, and lenses challenged with the slime-producing strain remained on the rabbit eye for longer time periods than those with a non-slime-producing strain. Bacteria associated with the contact lens may affect the retention of the lens on the rabbit cornea during experimental studies.

Animals↗

PubMed2004

Fatal influenza A infection with Staphylococcus aureus superinfection in a 49-year-old woman presenting as sudden death.

A fatal case of influenza A infection with Staphylococcus aureus superinfection in a previously healthy 49-year-old woman presenting as sudden, unexpected death is reported. Autopsy revealed severe necrotizing tracheobronchitis and hemorrhagic pneumonia. Microscopic examination of the trachea and bronchi showed mucosal necrosis and a dense lympho-monocytic infiltration of all layers. The lungs showed focal hemorrhagic pneumonia. No pathological changes were detectable in the myocardium. Influenza A virus was detected in bronchi and lung samples obtained during autopsy by the polymerase chain reaction (PCR) and bacterial superinfection with Staphylococcus aureus was shown by culturing from tracheal, bronchial and pulmonary swabs obtained during autopsy. PCR assays for the detection of Panton-Valentine leukocidin performed from all samples were negative. This case demonstrates the need for an interdisciplinary approach towards an organism-specific diagnosis of potentially infection-related deaths undergoing a medico-legal autopsy. With improved diagnostic possibilities such as PCR and DNA sequencing, forensic pathologists can, in close association with the field of microbiology, make a significant contribution to the detection of highly infectious agents which must be notified to the authorities. This will increase particularly the knowledge about the influence of these agents on sudden, unexpected deaths in outpatients.

Death, Sudden↗

PubMed2002

Minimal attachment killing (MAK): a versatile method for susceptibility testing of attached biofilm-positive and -negative Staphylococcus epidermidis.

Due to its ability to attach to polymeric surfaces Staphylococcus epidermidis is a common pathogen in chronic, medical device-associated infections. Attached S. epidermidis displays reduced susceptibility against a variety of antimicrobial substances, and little correlation between standard susceptibility test results and clinical outcome of antibiotic treatment is observed. In this study we established a new, versatile, and easy method of antimicrobial susceptibility testing for attached Staphylococcus epidermidis, suitable for both biofilm-negative and biofilm-positive attached bacteria using readily available equipment. For three biofilm-positive wild-type strains and their biofilm-negative mutants minimal attachment killing concentrations (MAK) of penicillin, oxacillin, vancomycin, and gentamicin were determined. Depending on strain and investigated antibiotics, a heterogeneous MAK (MAK(hetero)) could be differentiated from a homogeneous resistance (MAK(homo)), favoring a model of few persisters within attached cells under antibiotic treatment. For the biofilm-negative mutants, a lower MAK(homo) was detected than for the corresponding wild types for some of the tested antibiotics, which probably resulted from higher bacterial inocula of wild-type strains, whereas the MAK(hetero) were comparable for mutants and wild types for most of the tested antibiotics and strains. These data indicate that biofilm formation is not a necessary prerequisite for persistence of attached S. epidermidis cells under antibiotic treatment, which could explain therapeutic failure in foreign body-associated infections due to biofilm-negative S. epidermidis isolates. The highly individual resistance phenotypes of the investigated strains with different antibiotics suggests that MAK determination could help to predict the therapeutic outcome of foreign body-associated infections with both biofilm-positive and biofilm-negative S. epidermidis.

Anti-Bacterial Agents↗

PubMed2004

Primary sternal osteomyelitis due to community-associated methicillin-resistant Staphylococcus aureus: case report and literature review.

UNLABELLED: Primary sternal osteomyelitis is a rare infectious disease in paediatric patients. It has not been previously reported as caused by community-associated methicillin-resistant Staphylococcus aureus. We describe a 9-year-old boy with a painful erythematous mid-sternal mass diagnosed as primary sternal osteomyelitis. The aetiological agent was community-associated methicillin-resistant Staphylococcus aureus, carrying a type IV staphylococcal cassette chromosome for mec. CONCLUSION: Methicillin resistance should be taken into considered in patient with community-associated staphylococcal infections.

Child↗

PubMed2006

Comparison of community-acquired methicillin-resistant Staphylococcus aureus bacteremia to other staphylococcal species in a neonatal intensive care unit.

Hospital acquired infections including staphylococcal species are common in neonatal intensive care units. Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) was recently observed in our unit. The clinical and laboratory characteristics of all neonates with Staphylococcus aureus bacteremia during an 11-year period were retrospectively reviewed. Three groups of patients were compared: 1. Patients with CA-MRSA defined as MRSA-resistant only to beta-lactams, but sensitive to all other antibiotic groups and carried SCCmec IV. 2. Patients with multi-drug-resistant (MDR)-MRSA and 3. Patients with MSSA (methicillin-sensitive S. aureus). Forty-three neonates with documented S. aureus bacteremia were included. Of these 41 were preterm babies. Eleven infants had CA-MRSA, 20 had MDR-MRSA and 12 had MSSA bacteremia, the Panton-Valentine-Leukocidine gene (pvl-gene) was not present in any of these strains. Risk factors, clinical manifestations and laboratory tests were similar in all three groups studied. Although neonates infected with CA-MRSA were more premature and had more related diseases, the mortality rate was similar in all groups (9.1% in the CA-MRSA group). Skin infections, osteomyelitis or pneumatocele were not observed more frequently in the CA-MRSA group. We did not find significant differences in risk factors or outcomes in neonates in the three groups. One possible explanation for this observation is that the CA-MRSA outbreak strain did not contain the pvl-gene, which has been suggested to be a significant virulence factor.

Anti-Bacterial Agents↗

PubMed2004

Carcinogenic potential of tobacco tar-resistant Staphylococcus aureus in buccal cavity.

PURPOSE: The effects of cigarette smoking on the association between inflammation and cancer were studied, since some bacteria induce the production of tumor necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine and endogenous tumor promoter, in cells. METHODS: Bacteria from a gargled solution from the buccal cavity of 20 individuals were cultured in the presence of 4 mg/ml cigarette-smoke condensates. Although cigarette-smoke condensates inhibited growth of Staphylococcus aureus strongly and that of Staphylococcus warneri weakly, tobacco tar-resistant S. aureus and S. warneri were obtained. RESULTS: One tobacco tar-resistant S. aureus strain (Sa-TA10) induced expression of the TNF-alpha gene in both Bhas 42 cells (v-Ha-ras transfected BALB/3T3 cells) and human lung cancer cell line H226B, while one tobacco tar-resistant S. warneri (Sw-TA75) did not induce it significantly. Moreover, Sa-TA10 induced formation of transformed foci and soft-agar colony in Bhas 42 cells in cooperation with the v-Ha-ras gene. The results suggested that Sa-TA10 has carcinogenic potential, whereas Sw-TA75 does not. CONCLUSION: These data suggest that tobacco tar-resistant S. aureus, with carcinogenic potential, is present in the buccal cavity of some individuals, and that cigarette smoking simultaneously inhibits growth of most of the bacteria and selects carcinogenic bacteria.

3T3 Cells↗

PubMed2006

Nasal carriage of methicillin-resistant staphylococcus aureus in vascular surgery.

The purpose of this study was to determine the prevalence of nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) and to define risk factors allowing identification of high-risk patients for MRSA nasal carriage at admission to the vascular surgery unit. From March 23, 2004 to July 13, 2004, screening for nasal carriage of MRSA was conducted at admission to the vascular surgery unit and 1 week thereafter. To analyze risk factors for MRSA nasal carriage at admission to the vascular surgery unit, a case-control study was carried out in patients presenting colonization at the time of admission. A total of 308 patients underwent nasal screening for MRSA. Thirteen were colonized with MRSA (nine at admission and four acquired), i.e., 4.2% of patients. Methicillin-susceptible Staphylococcus aureus (MSSA) was found in 11.4% of patients who underwent screening. Six patients with MRSA infection were identified during the study period. The two patients who acquired infection were colonized at the time of admission. Arrival from another health-care facility and from another department was a significant risk factor for carriage of MRSA. The prevalence of nasal carriage in vascular surgery was 4.2%. Nasal screening is highly cost-effective since 60% of MRSA carriers were undetected using diagnostic specimens alone. French recommendations issued for cardiac and orthopedic surgery by the consensus conference on preoperative management of infectious risk on March 5, 2004, should be extended to vascular surgery.

Adult↗

PubMed2002

Selection of oxacillin resistance in oxacillin-sensitive Staphylococcus aureus through exposure to ciprofloxacin but not piperacillin/tazobactam.

Ten oxacillin-sensitive Staphylococcus aureus strains were grown on agar containing four times their ciprofloxacin MIC to determine if exposure to ciprofloxacin would increase their resistance to oxacillin. All strains grew on the ciprofloxacin-containing agar and subsequently grew on oxacillin-salt agar. The geometric mean MICs for oxacillin increased one- to sixteen-fold and remained elevated after ten passages on antibiotic-free agar. The mecA gene was not detected in any strain. There was no increase in oxacillin MICs when the bacteria were passaged on agar containing four times their MIC of piperacillin/tazobactam. Exposure of oxacillin-sensitive strains of Staphylococcus aureus to ciprofloxacin may increase their MICs to oxacillin.

Anti-Infective Agents↗

PubMed2002

Costs associated with a strict policy to eradicate methicillin-resistant Staphylococcus aureus in a Dutch University Medical Center: a 10-year survey.

Although the Dutch policy to eradicate methicillin-resistant Staphylococcus aureus (MRSA) is very strict compared to policies employed in other countries, it has proven to be successful epidemiologically (incidence of MRSA in the Netherlands, <0.5%). The present study was performed to investigate both the financial and the logistical consequences of this strict, so-called "search and destroy" policy in the Netherlands. The data were based on a 10-year survey (1991-2000) of screening, surveillance, and outbreaks at the University Medical Center Utrecht. The consequences of the policy were determined by a panel comprising physicians from the Department of Surgery, the Department of Medical Microbiology, Subdivision Hospital Hygiene and Infection Prevention, the Department of Pharmacy, and Household Services. The costs associated with the policy were also calculated, including those for additional (disposable) material, cultures, specific medication, decontamination, and closing of the wards. Over the course of the 10 years, implementation of the MRSA policy resulted in more than 2,265 lost hospitalization days. In addition, the wards had to be closed 48 times, 29 healthcare workers had to temporarily discontinue working, and 78,000 additional cultures had to be performed. The total cost reached 6 million Dutch guilders (euro 2,800,000). The financial and logistical consequences were then compared to those in a hypothetical situation without the "search and destroy" policy. In such a situation, the hospital would be faced with an increased incidence of MRSA, vancomycin intermediate-susceptible Staphylococcus aureus, and vancomycin-resistant enterococci. The costs associated with the use of alternative antibiotics, required in a scenario of high endemic-level MRSA, would be at least twice as high as the costs expended in the actual situation, thus demonstrating that a strict MRSA policy is financially worthwhile.

Academic Medical Centers↗

PubMed2002

Clinical study of 44 cases of Staphylococcus aureus meningitis.

In order to describe the clinical features and outcome of Staphylococcus aureus meningitis, the charts of 44 cases seen at one teaching hospital during a 20-year period were retrospectively reviewed. Staphylococcus aureus was the fifth most common cause of bacterial meningitis (10.2% of cases). There were 28 (63%) cases of postoperative meningitis and 16 (37%) of spontaneous meningitis. Patients with postoperative meningitis were younger than patients with spontaneous meningitis (mean age, 40.3 vs. 59.3 years; P=0.04) and had a lower frequency of community-acquired infection (32.1% vs. 93.8%; P<0.01), severe underlying diseases (28% vs. 87%; P<0.01) and associated staphylococcal infection (35% vs. 75%; P=0.01). The clinical presentation was similar in both groups, but patients with postoperative meningitis had a lower frequency of altered mental status (39% vs. 75%; P=0.02), meningeal signs (28% vs. 62%; P=0.02), petechial rash (0% vs. 18%; P=0.04), bacteremia (7% vs. 75%; P<0.01), and septic shock (0% vs. 44%; P<0.01). Most patients were treated with cloxacillin or vancomycin, with or without rifampicin, for a mean period of 22.5 days (range, 1-100 days). Overall mortality was 27%, and the mortality rate was higher for cases of spontaneous than postoperative meningitis (50% vs. 14%; P=0.01). Mortality correlated significantly with advanced age, spontaneous meningitis, altered mental status, and the presence of severe underlying diseases, bacteremia, and septic shock.

Anti-Bacterial Agents↗

PubMed2002

Diversity of Staphylococcus aureus strains isolated from two European regions with different prevalences of methicillin resistance.

The genodiversity of Staphylococcus aureus isolates from the Nottingham region of the United Kingdom was compared with isolates from the Freiburg region of Germany. The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) isolates was higher in Nottingham than in Freiburg. In patients from Nottingham hospitals, 80% of MRSA isolates were classical epidemic MRSA-15, but genotypic variants of epidemic MRSA-15 comprised 72% of isolates from Nottingham community-based patients. In contrast, MRSA isolates from Freiburg showed greater diversity, but 47% and 23% of isolates, respectively, belonged to two predominant MRSA genotypes found in isolates from both hospitalised and community-based patients. The results suggest that genodiversity becomes increasingly more confined in settings with a higher frequency and longer duration of MRSA prevalence.

Genetic Variation↗

PubMed2003

Molecular epidemiology of oxacillin-resistant Staphylococcus aureus in Brooklyn, New York.

Although oxacillin-resistant Staphylococcus aureus (ORSA) are well-established pathogens in many hospitals, the impact of infection control measures on the clonal distribution of ORSA is poorly defined. A citywide surveillance study of Staphylococcus aureus in Brooklyn, New York revealed that 36% of isolates were ORSA. Molecular typing showed that one strain (Cluster A) accounted for more than half of the isolates and was present in all 15 hospitals. In one hospital, a distinct strain (Cluster B) accounted for 20% of ORSA isolates. Infection control measures in this hospital significantly decreased the percentage of clinical isolates that belonged to Cluster B, but did not have an effect on the strain endemic to the city (Cluster A). Regional infection control strategies may need to be developed to limit the spread of the ORSA clone endemic to this area.

Anti-Bacterial Agents↗

PubMed2003

Emergence of a teicoplanin-resistant small colony variant of Staphylococcus epidermidis during vancomycin therapy.

Small colony variants of Staphylococcus aureus can cause persistent and recurrent infections. There are only a few reports of small colony variants of coagulase-negative staphylococci. Herein a case of infection with a teicoplanin-resistant small colony variant of Staphylococcus epidermidis is presented. The small colony variant was isolated from blood cultures of a patient with acute leukaemia and therapy-induced neutropenia who was treated with vancomycin for catheter-associated bloodstream infection. Despite removal of the catheter and adequate antibiotic therapy, the infection did not clear and the patient died 20 days after continuous antibiotic therapy.

Aged↗

PubMed2003

Obvious lack of association between dynamics of epidemic methicillin-resistant Staphylococcus aureus in central Europe and agr specificity groups.

During the past 8 years, changes in the prevalence and spread of different epidemic methicillin-resistant Staphylococcus aureus (MRSA) have been observed in central Europe, with the emergence of new strains possessing fewer resistance characters. This has also been demonstrated at the level of particular hospitals. Since variation in agr specificity type has been proposed as a possible reason for population dynamics in Staphylococcus aureus, the agr specificity groups of different epidemic MRSA strains were investigated by PCR using agr group-specific primers. Four of the "old" as well as two "new" epidemic strains exhibited agr specificity group I. One group of epidemic MRSA strains, which has been observed since the beginning of the 1990s, exhibited the agr specificity group II. Sequencing the variable part ( agrB-D-C) of the agr locus revealed only six relevant nucleotide changes within this region, with three of them modifying the Shine-Dalgarno sequence region of agrC. On the basis of the results obtained, it is proposed that the dynamics observed in the population of MRSA in Germany is not due to different agr group specificities in "old" and "new" epidemic clones.

Bacterial Proteins↗

PubMed2004

Long-term outcome and quality of care of patients with Staphylococcus aureus bacteremia.

To assess the long-term outcome and influence of clinical management of patients with Staphylococcus aureus bacteremia (SAB), 229 patients with blood cultures positive for Staphylococcus aureus between January 1997 and December 2000 were retrospectively identified and followed up. Risk factors, source of infection, treatment, clinical course, and outcome were recorded by chart review. For the assessment of 1-year survival, a questionnaire was sent to family doctors and government registration offices. Time of initial antibiotic therapy, duration of antibiotic treatment and performance of echocardiography were regarded as indicators of the quality of the clinical management of SAB. Among the 229 patients studied, 218 were evaluable for 1-year survival. Crude mortality after 1 year was 37.6% year. Within 30 days 43 (19.7%) patients had died, and 39 (17.9%) additional patients died thereafter. Using multivariate analysis, the following variables were associated with death: malignant disease (odds ratio [OR] 4.8; 95% confidence interval [CI], 2.6-8.9), pneumonia (OR, 3.6; 95%CI, 1.2-10.2), age >60 years (OR, 2.6; 95%CI, 1.5-4.5), and known source of infection (OR, 2.3; 95%CI, 1.3-4.1). Among 160 patients with a completely assessable treatment course 73 (46%) had received antibiotics for at least 14 days. A delay of antibiotic treatment of 1 day or more after microbiological diagnosis was observed in 28.3% of patients (i.e., 60 of 212 patients who received at least 1 dose of antibiotics). Echocardiography was performed in 101 (44.1%) cases. Overall, the findings indicate that standard guidelines for the management of SAB are followed only in part in clinical practice. In order to reduce the considerable mortality associated with SAB and to improve short- and long-term outcome, efforts should be made to increase adherence to recommendations.

Adult↗

PubMed2005

Low-level vancomycin resistance in Staphylococcus aureus--an Australian perspective.

Low-level vancomycin resistance in Staphylococcus aureus has emerged as a clinical problem over the past 8 years. The clinical relevance of this resistance has been questioned, and laboratory detection remains difficult and time consuming. There is, however, increasing evidence linking low-level vancomycin resistance with glycopeptide treatment failure in serious Staphylococcus aureus infections. Diagnostic laboratories and clinicians need to be aware of this resistance phenotype, to have procedures in place to detect the resistance, and to have strategies for managing patients with infections caused by resistant strains.

Aged↗