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Serology of Staphylococcus aureus infections using multiple antigens and serial serum samples.

The detection of antibody to S aureus in human serum can aid in the management of staphylococcal diseases [1]. RIAs and ELISAs can detect low levels of antibody and demonstrate increased antibody production during serious staphylococcal infections [2,3]. We compared four S aureus constituents--cell-wall peptidoglycan and teichoic acid, and extracellular alpha-toxin and nuclease--as antigens in a sensitive ELISA. The value of testing more than a single serum sample was also determined. Elevated IgG antibody to peptidoglycan, present in one or more serum samples of 13 (50%) of 26 patients with complicated bacteremia, was found to be the most sensitive test. All 26 patients had a significant IgG antibody response to peptidoglycan. Three (27%) of 11 patients with uncomplicated bacteremia had elevated levels of antibody to peptidoglycan in their serum, and seven (64%) showed a significant change in titer when serial serum samples were tested. Maximum detection rates for the other antigens in complicated and uncomplicated bacteremia were, respectively, 62% and 37% for teichoic acid, 38% and 37% for nuclease, and 54% and 13% for alpha-toxin. In single serum samples, the detection rate for all four antigens marginally improved the results, with detection rates of 62% and 36% for complicated and uncomplicated bacteremia, respectively. Cross-reactive antibody to peptidoglycan but not to the other three antigens was present in six (75%) of eight patients with long-standing subacute bacterial endocarditis due to either viridans streptococci or Staphylococcus epidermidis (data not shown).(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Bacterial↗

Antimicrobials as potential adjunctive agents in the treatment of biofilm infection with Staphylococcus epidermidis.

BACKGROUND: This study was performed to evaluate the interaction of erythromycin or rifampin with vancomycin against Staphylococcus epidermidis biofilms. METHODS: Biofilm-forming S. epidermidis strains (ATCC 35983, 35984) and polyurethane (PU) sheets were incubated for the formation of bacterial biofilms. Biofilms on PU sheets were treated with various antibiotic regimens. The number of viable bacteria on the sheets was counted. RESULTS: While erythromycin or vancomycin alone did not significantly reduce the biofilm bacterial concentration, the combination of vancomycin and erythromycin resulted in a clear reduction in bacterial concentration compared with the single agents. Rifampin was the most active single agent against biofilm-forming S. epidermidis, while the combination of rifampin and vancomycin showed further reduction in bacterial concentration. CONCLUSIONS: These data suggest that the combination of erythromycin or rifampin and vancomycin is more effective than vancomycin alone in the treatment of biofilm infections caused by biofilm-forming S. epidermidis.

Biofilms↗

Hydrophobized wound dressing in the treatment of experimental Staphylococcus aureus infections in the young pig.

Standard wounds (2 X 2 cm) produced with a dermatome, followed by a thermal injury in four-week-old piglets were inoculated with Staphylococcus aureus strains. Infection developed in each wound within a few days. Wounds were then dressed separately with cellulose dressing or the same dressing substituted with a hydrophobic ligand (stearylic acid). Other wounds were treated with active charcoal dressing (ActisorbR) or DebrisanR. Signs of healing, including vanishing of inflammation around the wounds, were noticed already on days 3 and 4 in the wounds treated with the hydrophobized dressing, and no infection was noticed in such wounds on days 5 to 6, while all other wounds were still showing signs of infection. It is concluded that active removal of S. aureus cells in the hydrophobized dressing probably decreases the number of multiplying bacteria in wounds, causing a rapid onset of wound healing.

Animals↗

Comparison of length of hospital stay for patients with known or suspected methicillin-resistant Staphylococcus species infections treated with linezolid or vancomycin: a randomized, multicenter trial.

STUDY OBJECTIVE: To compare hospital length of stay (LOS), weekly discharges, and days of antibiotic treatment with linezolid (intravenous with oral follow-up) and vancomycin (intravenous only). DESIGN: Multinational, randomized, phase III trial. SETTINGS: Hospitals in North America, Latin America, and Europe. PATIENTS: Four hundred sixty hospitalized patients with infections of known or suspected methicillin-resistant Staphylococcus species. INTERVENTION: Administration of linezolid or vancomycin. MEASUREMENTS AND MAIN RESULTS: For linezolid recipients, median LOS was 5 and 8 days shorter (p=0.05 and 0.003) in the complicated skin and soft tissue infection intent-to-treat (230 patients) and clinically evaluable (144) samples, and slightly but not significantly shorter in the overall intent-to-treat (460) and clinically evaluable (254) samples. In all samples, linezolid recipients had more discharges in the first week of treatment and fewer days of intravenous therapy than vancomycin recipients. CONCLUSION: Our results support linezolid's ability to reduce medical resource use.

Acetamides↗

Very severe thrombocytopenia and fragmentation hemolysis mimicking thrombotic thrombocytopenic purpura associated with a giant intracardiac vegetation infected with Staphylococcus epidermidis: role of monocyte procoagulant activity induced by bacterial supernatant.

The pathogenesis of very severe thrombocytopenia in bacterial endocarditis is uncertain. We report a 50-year-old male with platelet counts < 10 x 10(9)/l and fragmentation hemolysis complicating Staphylococcus epidermidis pacemaker endocarditis with a giant vegetation. Antibiotics, corticosteroids, high-dose intravenous gammaglobulin, and plasmapheresis (for initially-suspected thrombotic thrombocytopenic purpura) failed to produce significant platelet count increase. However, therapeutic-dose heparin anticoagulation was associated with a platelet count increase from <10 to approximately 40 x 10(9)/l, with parallel reduction in thrombin-antithrombin complexes (from 8.9 to 3.5 microg/l), facilitating surgical intervention. The thrombocytopenia promptly resolved following surgical removal of the vegetation. Culture supernatant from S. epidermidis isolated from the patient's blood induced monocytes to express procoagulant activity (assessed by factor Xa generation) equivalent to lipopolysaccharide (1 microg/ml), with half-maximal activation seen with culture supernatant diluted to 1:12,800. These data are consistent with previous animal models of endocarditis demonstrating staphylococci-induced procoagulant changes in monocytes. This case demonstrates that heparin anticoagulation can be therapeutic in infective endocarditis-associated severe thrombocytopenia in a non-bleeding patient, and that such therapy may ameliorate the platelet count enough to permit surgical intervention.

Blood Coagulation↗

Epidemiological study of an outbreak of infection with Staphylococcus aureus resistant to lincosamides and streptogramin A in a French hospital.

A significant increase in the incidence of isolates of methicillin-resistant Staphylococcus aureus (MRSA), that were also resistant to lincosamides and streptogramin A (LSA-MRSA), was observed in a French university hospital. Twenty-seven isolates from the outbreak were characterised, including 17 isolates from a plastic surgery ward and six control strains of MRSA. The strains were examined by antibiotyping and biotyping, and by three molecular methods: plasmid analysis, ribotyping and insertion sequence (IS) typing with IS256 sequence as a probe. Antibiotyping (five antibiotypes) was discriminatory because of the uncommon resistance phenotype of the epidemic strain. Biotyping (three biotypes), DNA plasmid analysis (four profiles) and ribotyping (two profiles) were poorly sensitive, in contrast to IS-typing (12 profiles). By the latter method, a coefficient of similarity (percentage similarity) compared to the predominant IS profile was calculated. Strains with a coefficient of similarity > or = to 82% were considered as highly related to the epidemic strain, while those with a coefficient of similarity < or = to 40% were regarded as distant. Results obtained with the five markers confirmed that an outbreak of hospital infection had occurred in the plastic surgery ward, with spread of the epidemic strain throughout the hospital.

Adult↗

The lantibiotic lacticin 3147 produced in a milk-based medium improves the efficacy of a bismuth-based teat seal in cattle deliberately infected with Staphylococcus aureus.

A preparation of the bacteriocin lacticin 3147 (prepared from a demineralized whey protein fermentation liquor) was combined as a powder with a bismuth-based intramammary teat seal and evaluated for its potential as an antimicrobial in non-lactating cows. The lacticin/teat seal formulation enabled significant bacteriocin release from the seal without the requirement for a surfactant. Studies in vivo in lactating cows demonstrated that this formulation was effective in reducing bacterial recoveries (approximately 20-fold) from teats deliberately inoculated with Staphylococcus aureus after infusion. Moreover, this formulation also significantly reduced the numbers of Staph. aureus recovered from teats that were exposed to the challenge bacterium before the infusion of the teat seal preparation. The powdered preparation of lacticin 3147 did, however, cause some teat irritation as evidenced by associated rises in somatic cell count (SCC). However, this effect was short-lived and when the mean SCC readings pre-infusion and the final two readings post-infusion were compared, there was no significant difference in the immunological acceptance between treatments.

Animals↗

The influence of calcium mupirocin nasal ointment on the incidence of Staphylococcus aureus infections in haemodialysis patients.

Mupirocin was used in haemodialysis patients in an attempt to eradicate nasal carriage of Staphylococcus aureus and to prevent infection caused by this microorganism. The effectiveness of calcium mupirocin as a 2% nasal ointment OB2 (16 patients for 104 patient-months) was compared to that of placebo (18 patients for 147 patient-months) in a double-blind study. Mupirocin or placebo were applied in both anterior nares thrice daily for 2 weeks and subsequently three times weekly for a total of 9 months. During therapy, S. aureus was recovered from only 6% of the nasal cultures in the mupirocin group compared to 58% in the placebo group (P less than or equal to 0.01). Only one S. aureus infection was documented in the mupirocin group compared to six in the placebo group (P less than or equal to 0.05). The S. aureus strain causing the single infection in the mupirocin group was of a different phage type to that of the original nasal strain. In contrast, at least four of the six strains causing infection in the placebo group were of similar phage type to the original nasal strain. All S. aureus isolates remained mupirocin sensitive (MIC less than or equal to 1 mg/l). In conclusion, mupirocin nasal ointment was effective in eradicating nasal carriage of S. aureus and in preventing S. aureus infections in patients on haemodialysis.

Administration, Intranasal↗

Nosocomial infection by Staphylococcus haemolyticus and typing methods for epidemiological study.

A patient with chronic myelogenous leukemia became colonized with a Staphylococcus haemolyticus strain and experienced a septic episode caused by this strain during a cytostatic course. The strain was multiply resistant to antibiotics; the MIC and MBC of vancomycin were 2 and 4 mg/liter, and the MIC and MBC of teicoplanin were 4 and 16 mg/liter, respectively. We performed a surveillance study on the carriage of S. haemolyticus in medical and nursing staff of the hospital ward where the patient was treated. S. haemolyticus was isolated from 18 sites on 12 of the 39 people tested. A number of typing methods were performed in order to investigate the possible relationships among the isolates. Methods used were immunoblotting of staphylococcal peptides, plasmid analysis, restriction fragment length polymorphism of chromosomal DNA, and pulsed-field gel electrophoresis of total DNA. Compared with the immunoblotting technique, the molecular methods were more discriminative. The strain colonizing the patient showed a consistent pattern by all typing methods during isolation. When the immunoblot technique was used, similar patterns were found with isolates from hospital staff and isolates from unrelated sources. With the molecular techniques, no evidence of a local spread of the patient's strain was found. However, plasmid profiles and restriction fragment length polymorphism and pulsed-field gel electrophoresis patterns showed that S. haemolyticus isolates collected from hospital ward personnel were related, which was not the case with isolates collected from unrelated sources. Restriction fragment length polymorphism analysis was more discriminative when IS431 was used as a DNA probe instead of a probe based on the 16S rRNA gene. S. haemolyticus, as in this case, may develop resistance to vancomycin and teicoplanin. These antibiotics are considered the last-resort drugs for the therapy of nosocomial gram-positive infections. Thus, local spread of staphylococci resistant to these drugs is an important problem, which should be prevented by strict hygienic measures and antibiotic policy.

Base Sequence↗

Area under the inhibitory curve and a pneumonia scoring system for predicting outcomes of vancomycin therapy for respiratory infections by Staphylococcus aureus.

Treatment factors predictive of clinical and microbiological outcomes and the relationship between a pneumonia scoring system and clinical outcomes in vancomycin-treated patients with a Staphylococcus aureus-associated lower-respiratory-tract infection (LRTI) were studied. A computer database review identified patients for whom S. aureus was isolated from a respiratory-tract specimen between January 1 and December 31, 1998, and who had antimicrobials ordered within 72 hours of isolation of that organism. Through further review of individual patient charts, this group was restricted to those treated with vancomycin for a documented S. aureus-associated LRTI. Classification-and-regression-tree (CART) modeling was performed to determine which clinical variables were correlated with clinical outcomes and microbiological outcomes. Median changes in clinical pneumonia scores from baseline in two patient groups (those with clinical success and those with clinical failure) were compared. Seventy patients met the study criteria. CART modeling found that both outcomes were associated with area under the inhibitory curve (AUIC). The pneumonia scoring system was predictive of eventual clinical success as early as day 3 of treatment; having at least a 4-point decrease in the pneumonia score by day 3 was correlated with an 87% clinical success rate. Both AUIC and a pneumonia scoring system were useful for predicting clinical and microbiological outcomes of vancomycin therapy in patients with LRTIs caused by S. aureus.

Adult↗

Efficacy of teicoplanin and autoradiographic diffusion pattern of [14C]teicoplanin in experimental Staphylococcus aureus infection of joint prostheses.

Prosthesis infections are difficult to cure. Infection with methicillin-resistant staphylococci is becoming more common in patients with orthopedic implants. Using a recently developed model of methicillin-resistant Staphylococcus aureus (MRSA) infection of a knee prosthesis, we compared the efficacies of teicoplanin and vancomycin. [14C]teicoplanin diffusion in this model was also studied by autoradiography. A partial knee replacement was performed with a silicone implant fitting into the intramedullary canal of the tibia, and 10(7) CFU of MRSA was injected into the knee. Treatment with teicoplanin or vancomycin (20 or 60 mg/kg of body weight, respectively, given intramuscularly twice daily) was started 7 days after inoculation and was continued for 7 days. The teicoplanin and vancomycin MICs for MRSA were 1 microg/ml. Mean peak and trough levels in serum were 39.1 and 23.5 microg/ml, respectively, for teicoplanin and 34.4 and 18.5 microg/ml, respectively, for vancomycin. Fifteen days after the end of therapy, the animals were killed and their tibias were removed, pulverized, and quantitatively cultured. Teicoplanin and vancomycin significantly reduced (P < 0. 05) the bacterial density (2.7 +/- 1.3 and 3.3 +/- 1.6 log10 CFU/g of bone, respectively) compared to those for the controls (5.04 +/- 1.4 log10 CFU/g of bone). The bacterial covents of teicoplanin- and vancomycin-treated rabbits were comparable. The [14C]teicoplanin autoradiographic diffusion patterns in rabbits with prostheses, two of which were uninfected and two of which were infected, were studied 15 days after inoculation. Sixty minutes after the end of an infusion of 250 microCi of [14C]teicoplanin, autoradiography showed that in the infected animals, the highest levels of radioactivity were located around the prosthesis and in the periosteum, bone marrow, and trabecular bone. Radioactivity was less intense in epiphyseal disk cartilage, femoral cartilage, articular ligaments, and muscles and was weak in compact bone. A similar distribution pattern was seen in uninfected rabbits. Thus, teicoplanin may represent an effective alternative therapy for the treatment of these infections.

Animals↗

Low-dose infectivity of Staphylococcus aureus (SMH strain) in traumatized rat tibiae provides a model for studying early events in contaminated bone injuries.

Animal models of post-traumatic acute osteomyelitis (OM) that closely mimic human scenarios, including infection prophylactic procedures such as debridement and lavage, may provide a better understanding of OM. We contaminated mechanically traumatized rat tibiae (n = 69) with various doses of a Staphylococcus aureus strain (SMH) known to cause human OM and then performed curettage and lavage. Tibiae were harvested 24 h after lavage for assessment of bacterial load and determination of minimal infective doses for 50% (ID50) and 95% (ID95) of rats. Some experiments varied tibial harvest time after lavage (n = 10); for progressive infection, tibiae were evaluated at 7 and 15 days after contamination (n = 17 for each time point). At 24 h after contamination, the ID50 was 1.8 x 10(3) CFU, and the ID95 was 9.2 x 10(3) CFU. Tibial bacterial loads did not increase with inocula greater than the ID95. Lavage removed many bacteria from bone, but it did not prevent subsequent infection or disease. At 15 days after contamination, most tibiae (14 of 17) were infected, with macroscopic and radiological signs of established OM. This newly described rat OM model, with a low ID95 despite prophylactic curettage and lavage, closely mimics events in contaminated human bone injuries. This situation will allow study of early factors in contaminated bone injuries, including clinical interventions that may reduce infection and prevent disease.

Animals↗

Preventing methicillin-resistant Staphylococcus aureus infection in joint replacements: a new problem requiring old solutions.

AIM: To analyse prospectively the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) colonisation and infection in wounds and surgical vacuum drains. METHODS: The study group was 100 patients undergoing primary total hip and total knee replacements in a single orthopaedic unit. RESULTS: Two patients had undergone revision joint surgery within the same unit in the year prior to the study due to confirmed MRSA infection. Following a change in the unit's infection control policy, joint replacement surgery continued within the unit even when there were cases of active MRSA present on the same ward. None of the patients undergoing joint replacement grew MRSA in their postoperative drain tips or wound cultures. CONCLUSIONS: It is safe to continue with joint replacement surgery when other patients on the same ward have active MRSA as long as strict infection control measures are followed as outlined.

Antibiotic Prophylaxis↗

Changes in the serum protein binding of vancomycin in patients with methicillin-resistant Staphylococcus aureus infection: the role of serum alpha 1-acid glycoprotein levels.

The relationship between albumin or alpha 1-acid glycoprotein (AAG) levels and vancomycin (VCM) protein binding was studied in 44 serum samples from the 10 patients with methicillin-resistant Staphylococcus aureus (MRSA) infection receiving VCM therapy. Eighty serum samples from 80 healthy subjects were used as a control for albumin and AAG levels. The protein binding percentage of VCM in the serum of the patients varied widely from 27 to 62%. The mean albumin level (34 g/L) was significantly lower than that in healthy subjects (46 g/L). There was no correlation between the binding percentage and serum albumin level in the patients (r = -0.25, p > 0.1). The mean AAG level in the patients (1.51 g/L) was significantly higher than in healthy subjects (0.59 g/L). There was a significant correlation between the binding percentage of VCM and serum AAG level (r = 0.63, p < 0.001). The binding percentage of VCM individual patients changed in parallel to serum AAG and C-reactive protein (CRP) levels, which is a useful marker of acute phase response. There were also significant correlations between AAG, albumin, and CRP levels. The present results indicate that the increased AAG level in serum of the patients appeared to have a significant effect on the protein-binding characteristics of VCM.

Aged↗

Methicillin-resistant Staphylococcus aureus infection does not adversely affect clinical outcome of lower extremity amputations.

Methicillin-resistant Staphylococcus aureus was first identified in isolation in Europe during the 1960's. Now widespread throughout the world, infection with this organism has emerged as a major problem in surgical practice. However, it remains debatable whether MRSA is more virulent than methicillin-susceptible strains. We have reviewed our most recent 4-year experience of lower extremity amputations to examine the influence of MRSA and non-MRSA infection on clinical outcome. During the past 4 years, 165 patients underwent lower extremity amputation for SVS/ISCVS category III acute limb ischemia and grades II and III chronic limb ischemia. Forty-five had documented MRSA infection, while 57 patients had documented infection with other flora. All patients were treated with appropriate sensitivity-specific antibiotics and aggressive wound care. No significant differences were noted in the level of primary amputation required by the two groups. Similarly, no significant differences were noted in either number of revisions or revision to higher-level amputation, time to heal, hospital length of stay, or 30-day morbidity and mortality rates. Our results demonstrate that MRSA infection does not adversely affect clinical outcome in patients undergoing lower extremity amputations. Appropriate treatment of infections with sensitivity-specific antibiotics, thorough wound debridement, and aggressive wound monitoring should be routine in all patients, regardless of bacterial flora.

Aged↗

Protective role of sialophorin (CD43)-expressing cells in experimental Staphylococcus aureus infection.

Sialophorin (CD43) is a major surface mucin on most hematopoietic cell lineages, including phagocytes. Defects of CD43 expression occur in Wiskott-Aldrich syndrome, a disease characterized by susceptibility to pyogenic infections. In a newly established rat model of septic Staphylococcus aureus arthritis, we have investigated the role of CD43-expressing cells in the progression of the disease. A single injection of a monoclonal antibody specific for CD43 induced a highly erosive course of arthritis and increased mortality in animals exposed to a suboptimal dose of bacteria. Our results demonstrate that sialophorin-expressing cells play a protective role in the early stage of staphylococcal infection.

Animals↗