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Clarithromycin for experimental Staphylococcus aureus keratitis.

PURPOSE: Clarithromycin, a macrolide antibiotic not previously tested against the common causes of bacterial keratitis, was analyzed for its effectiveness in reducing the number of viable bacteria in a Staphylococcus keratitis model. An in vivo comparison of the effectiveness of clarithromycin to erythromycin, minocycline, and tetracycline for three strains of Staphylococcus aureus was done. METHODS: Rabbit eyes were intrastromally injected with 100 colony forming units of one of three strains of S. aureus. Two strains were methicillin-sensitive (ATCC 25923 and MSSA 309) and one strain methicillin-resistant (COL). Eyes were treated every 30 minutes with 0.3% clarithromycin, erythromycin, tetracycline, or minocycline from 4 to 9 hours postinfection. The number of colony forming units (CFU) per cornea in all eyes was determined at 10 hours postinfection. RESULTS: Vehicle-treated and untreated eyes (controls) contained over 6 logs of CFU per cornea, a value significantly higher than any of the antibiotic-treated eyes (P < or = 0.0001). Clarithromycin or erythromycin therapy significantly decreased the number of CFU per cornea by approximately 5 logs in the eyes infected with the methicillin-sensitive strains and by approximately 4 logs in the eyes infected with the methicillin-resistant strain. Tetracycline and minocycline were also successful in treating these strains, but overall showed less effectiveness than clarithromycin and erythromycin. CONCLUSIONS: Clarithromycin proved to be an effective ocular medication for the therapy of experimental S. aureus keratitis. The effectiveness of clarithromycin in this model and its known effectiveness for a variety of bacterial pathogens suggests a role for this drug as a useful ocular antibiotic.

Animals↗

The effectiveness of tobramycin and Ocuflox in a prophylaxis model of Staphylococcus keratitis.

PURPOSE: To determine the effectiveness of prophylactic antibiotic treatment prior to intra-corneal infection with Staphylococcus aureus. METHODS: One topical drop of Tobrex (0.3% tobramycin), tobramycin (0.3%) in the Tobrex vehicle with 0.05% dodecyl maltoside (DDM)/4.0% hydroxypropylmethycellulose (HPMC), Ocuflox (0.3% ofloxacin) or DDM/HPMC vehicle were applied to rabbit eyes at one or five hours prior to injection of bacteria. Approximately 500 colony-forming units (CFU) of S. aureus strain 8325-4 were injected into the corneal stroma. Rabbits were sacrificed five hours after infection and corneal homogenates were cultured to determine the number of colony forming units (CFU) per cornea. RESULTS: Rabbits treated at five hours prior to infection with tobramycin-DDM/HPMC reduced the bacterial load by approximately 2.4 log CFU/cornea as compared to the untreated control (3.47 +/- 0.98 vs. 5.71 +/- 0.14 log CFU/cornea, respectively; P = 0.0010); however, Ocuflox, Tobrex, or DDM/HPMC vehicle did not significantly reduce the log CFU (P >or= 0.4837). Rabbits treated at 1 hour prior to infection with Ocuflox or tobramycin-DDM/HPMC had significantly reduced CFU/cornea (1.31 +/- 0.86 and 0.48 +/- 0.31 log CFU/cornea, respectively) as compared to the untreated group (5.71 +/- 0.14 log CFU/cornea; P or= 0.2312). CONCLUSIONS: This pre-treatment model of Staphylococcus keratitis quantitatively measured the prophylactic effectiveness of topical antibiotic formulations. An important finding was that a tobramycin-DDM/HPMC formulation was highly effective as a prophylactic medication.

Animals↗

Design and evaluation of specific PCR primers for rapid and reliable identification of Staphylococcus xylosus strains isolated from dry fermented sausages.

Rapid and reliable identification of Staphylococcus xylosus was achieved by species-specific PCR assays. Two sets of primers, targeting on xylulokinase (xylB) and 60 kDa heat-shock protein (hsp60) genes of S. xylosus, respectively, were designed. Species-specificity of both sets of primers was evaluated by using 27 reference strains of the DSM collection, representing 23 different species of the Staphylococcus genus and 3 species of the Kocuria genus. Moreover, 90 wild strains isolated from different fermented dry sausages were included in the analysis. By using primers xylB-F and xylB-R the expected PCR fragment was obtained only when DNA from S. xylosus was used. By contrast, amplification performed by using primers xylHs-F and xylHs-R produced a single PCR fragment, of the expected length, when DNA from S. xylosus, S. haemolyticus, S. intermedius and S. kloosii were used as template. Nevertheless, AluI digestion of the xylHs-F/xylHs-R PCR fragment allowed a clear differentiation of these 4 species. The rapidity (about 4 h from DNA isolation to results) and reliability of the PCR procedures established suggests that the method may be profitably applied for specific detection and identification of S. xylosus strains.

Bacterial Proteins↗

Rapid and reliable identification of Staphylococcus equorum by a species-specific PCR assay targeting the sodA gene.

Rapid and reliable identification of Staphylococcus (S.) equorum was achieved by species-specific PCR assays. A set of primers targeting the manganese-dependent superoxide dismutase (sodA) gene of S. equorum was designed. Species-specificity of the primer set was evaluated by using a total of 112 strains (including 27 reference strains of the DSM collection), representing 26 different species of the genus Staphylococcus, 3 species of the genus Kocuria, and different strains of Macrococcus caseolyticus. By using primers SdAEqF and SdAEqR the expected PCR fragment was obtained only when DNA from S. equorum strains was used as template. The rapidity (about 4 h from DNA isolation to results) and reliability of the PCR procedures established suggests that the method may be profitably applied for specific detection and identification of S. equorum strains.

Bacterial Proteins↗

Treatment with Hypericum perforatum L. does not trigger decreased resistance in Staphylococcus aureus against antibiotics and hyperforin.

Most scientific investigations of Hypericum perforatum L. (Saint John's wort) concentrated on its antidepressant activity. Only recently, its antibacterial activity against multiresistant Staphylococcus aureus led to speculations regarding the use of hyperforin, an antibacterial principle of hypericum, as an antibiotic. In the present investigation, we show that Staphylococcus aureus is able to acquire a resistance against hyperforin which did not lead to a cross resistance against clinically used antibiotics. Resistance development does not take place, however, at concentrations as low as they are found in human blood plasma during antidepressant treatment with 900 mg Hypericum extract/day.

Bridged Bicyclo Compounds↗

Staphylococcus aureus in commercial laying flocks: incidence and characteristics of strains isolated from chicks, pullets and hens in an integrated commercial enterprise.

1. Staphylococcus aureus was able to colonise the surface of chicks as young as 1 d old. 2. The organism was detected in a hatchery particularly in the debris from the hatchers and on the working surfaces at the sexing and vaccination areas. 3. The degree of surface colonisation of chicks and pullets was low during rearing but rose to a maximum at mid-lay (50 weeks) when the organism was readily detectable on almost all hens. 4. Strains of Staphylococcus aureus were characterised by phage-typing and cultural characteristics and found to belong predominantly to poultry phage group B2 of Gibbs et al. (1978a). 5. A detailed description of poultry phage group B2 strains is presented; strains of this type can be considered as a second "live-bird" ecovar when compared with Staph. aureus var. gallinae of Witte et al. (1977).

Animals↗

Synercid plus vancomycin for the treatment of severe methicillin-resistant Staphylococcus aureus and coagulase-negative staphylococci infections: evaluation of 5 cases.

Synercid (quinupristin/dalfopristin), the first semi-synthetic injectable streptogramin, is a promising alternative to glycopeptides against many Gram-positive multiresistant bacteria. Vancomycin is still considered an effective agent for the treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections but therapeutic failures with glycopeptides have been observed, even for the treatment of infections caused by S. aureus strains sensitive to vancomycin. Synercid, in combination with a glycopeptide, may address this problem without causing significant side effects due to the different toxicity patterns of the 2 antimicrobials. This study reports our experience with the combination of Synercid and vancomycin in 5 patients with severe infection caused by MRSA or methicillin-resistant coagulase-negative Staphylococcus.

Abdominal Abscess↗

Primary cultures of human chondrocytes are susceptible to low inocula of Staphylococcus aureus infection and undergo apoptosis.

Hypocellularity after joint infection has been attributed to the cytotoxic effects of pus, which can cause necrosis of chondrocytes. In this study, primary cultures of human chondrocytes lost their viability and underwent necrosis rapidly with high inocula of Staphylococcus aureus infection. Chondrocytes were shown to undergo apoptosis with low inocula of Staphylococcus aureus or their culture ultrafiltrate. These findings further support the hypothesis that residual bacterial toxins or triggered apoptotic processes in chondrocytes participate in the pathogenesis of post-infectious arthropathy.

Apoptosis↗

Staphylococcus aureus meningitis: experience with cefuroxime treatment during a 16 year period in a Danish region.

Staphylococcus aureus is a rare cause of bacterial meningitis and there is no consensus on antibiotic treatment. Nafcillin is a common choice in countries where it is approved and marketed. High-dose cefuroxime has been the systemic treatment used in the study region, and a retrospective record review was conducted to determine its clinical efficacy. Cases of bacterial meningitis during 1984-1999 in the County of North Jutland, Denmark (approx. 490000 inhabitants), were identified in a regional bacteriology register. Inclusion of a case required either growth of S. aureus from > or = 2 specimens of cerebrospinal fluid (CSF), 1 positive CSF specimen with a CSF leucocyte count > 10(8)/l or 1 positive CSF specimen with a concurrent positive blood culture. A diagnosis of brain abscess required growth of S. aureus from aspirated pus. Staphylococcus aureus meningitis was confirmed in 45 patients, and 5 additional patients had a brain abscess. 44 cases were nosocomial (mortality 16%) and 6 were community acquired (mortality 83%). None of the isolates was methicillin resistant and 6 were penicillin susceptible. Intraventricular antibiotic treatment was given to 28 patients, systemic therapy included cefuroxime in 32 patients (64%) as either a primary or secondary choice, 6 (12%) were treated with penicillin G, 10 (20%) with penicillinase-resistant penicillin and 2 (4%) with cephalothin. Among 31 nosocomial cases treated systemically with cefuroxime the mortality was 10% (95% exact confidence limits 2-26%). In conclusion, cefuroxime seems to be a valid choice for S. aureus meningitis in the nosocomial setting.

Adolescent↗

Splenic abscess due to Staphylococcus lentus: a rare entity.

Splenic abscess is an unusual condition, most often seen in immunocompromised patients. Splenic abscesses are due to aerobic and anaerobic bacteria, fungi and parasites, Staphylococcus aureus being the most common aetiological agent. We describe a case of splenic abscess in an alcoholic patient due to Staphylococcus lentus, a rare aetiology included in the microbiology of this disease.

Abscess↗

Staphylococcus saprophyticus causing native valve endocarditis.

Coagulase negative staphylococci are a rare cause of native valve endocarditis. Staphylococcus saprophyticus is a coagulase-negative Staphylococcus infrequently reported as a human pathogen, and most of the cases reported are urinary tract infections. We describe a case of native valve endocarditis attributed to this organism. The patient needed valve replacement due to heart failure.

Aortic Valve↗

Fatal sepsis and necrotizing pneumonia in a child due to community-acquired methicillin-resistant Staphylococcus aureus: case report and literature review.

Pediatric deaths due to community-acquired methicillin-resistant Staphylococcus aureus are rare. We describe the case of 2-y-old boy with fever and cough followed by comatose state with deteriorated respiration; the boy died of severe sepsis and necrotizing pneumonia. The etiological agent was community-acquired methicillin-resistant Staphylococcus aureus, carrying a type IV staphylococcal mecA gene cassette and the Panton-Valentine Leukocidin gene.

Bacterial Proteins↗

Acute non-purulent inflammatory arthropathy associated with Staphylococcus aureus abscess.

Case 1. A 20-year-old woman presented 4 weeks post-partum with widespread symmetrical inflammatory polyarthropathy with marked synovitis. Investigations revealed grossly raised CRP with negative immunology screen. A few days before presentation she saw her general practitioner with left-sided mastitis, which then developed into a Staphylococcus breast abscess. Surgical drainage of this led to almost immediate resolution of the joint complaints and return of CRP to normal. Case 2. A 27-year-old man developed widespread symmetrical inflammatory arthropathy. A few days prior to this he had developed folliculitis with a furuncle on his neck. Swab grew Staphylococcus aureus. His arthritis settled immediately following spontaneous drainage of his abscess and a full course of antibiotic. The pathogenic mechanism is unclear but could be toxin-mediated.

Abscess↗

Screening for methicillin resistance in Staphylococcus aureus and coagulase-negative staphylococci: an evaluation of three selective media and Mastalex-MRSA latex agglutination.

Laboratory confirmation of MRSA is important for the implementation of infection control; conventional screening culture methods take up to five days for confirmation. The purpose of this study is to ascertain the efficiency of three selective media for growth of methicillin-resistant Staphylococcus aureus (MRSA) before and after enrichment in salt broth, and to evaluate the Mastalex-MRSA latex agglutination kit for detection of methicillin resistance. Screening swabs were collected from 63 patients, yielding 125 S. aureus isolates and 40 coagulase-negative staphylococcus (CNS) isolates. Selective media used were mannitol salt agar (MSA), Baird-Parker agar with ciprofloxacin (BPC) and bromocresol purple (BCPA). Polymerase chain reaction (PCR) for mecA gene detection was used as the reference standard for evaluation of the Mastalex-MRSA assay, which was also evaluated on colonies of S. aureus from the selective media. No significant difference was found in efficiency of MRSA isolation among the selective media. Pre-enrichment in the salt broth did not enhance isolation of MRSA. Methicillin-sensitive S. aureus and CNS were significantly inhibited in all selective media (P<0.05). Only BPC significantly selected out methicillin-resistant CNS (P<0.01). Mastalex-MRSA was 97% specific and sensitive for the detection of MRSA. It was 65% sensitive and 100% specific in detecting methicillin resistance in CNS. In conclusion, all selective media performed equally well (MRSA isolation rate of approximately 80%). Mastalex-MRSA provided rapid and reliable detection of MRSA from selective media, reducing the time required for confirmation of this organism.

Agglutination Tests↗

Staphylococci in animals: prevalence, identification and antimicrobial susceptibility, with an emphasis on methicillin-resistant Staphylococcus aureus.

Staphylococci form part of the normal flora of man and a wide variety of animals. Some staphylococcal species also cause a wide variety of pyogenic infections. The main pathogenic staphylococcus in man is Staphylococcus aureus. Increasingly, methicillin-resistant S. aureus (MRSA) has been a problem in the healthcare setting. Reports of MRSA in animals have been infrequent; however, evidence suggests that MRSA is being isolated increasingly from animals, in particular household pets and other companion animals. This article reviews the prevalence of pathogenic staphylococci in animals, with an emphasis on the emergence of MRSA as a possible animal pathogen.

Animals↗

Methicillin-resistant Staphylococcus aureus: acquisition and risk of death in patients in the intensive care unit.

OBJECTIVE: To evaluate the risk of patients in intensive care units (ICU) of becoming infected with methicillin-resistant Staphylococcus aureus (MRSA) and to assess the mortality during a six week follow-up period, compared with patients who developed methicillin-sensitive S. aureus (MSSA) infection. DESIGN: Point prevalence survey. SETTING: 1417 ICU in 17 Western European countries. SUBJECTS: 10038 patients in ICU who were part in the EPIC (European Prevalence of Infection in Intensive Care) Study. MAIN OUTCOME MEASURES: Prevalence of MRSA and MSSA ICU-acquired infections, risk factors, and mortality. RESULTS: On the study day 21% of patients had ICU-acquired infections. The most commonly reported pathogen was Staphylococcus aureus (30%). Overall, 60% of strains of S. aureus were resistant to methicillin (with a wide intercountry variation). The most commonly reported MRSA infections were pneumonia and lower respiratory tract infections. The most important risk factor for MRSA was the length of stay in the ICU. MRSA infection reduced the chance of survival, particularly when it was found in lower respiratory tract infections: the risk of mortality was three times higher in patients with MRSA than in those with MSSA. CONCLUSION: Patients in ICU are at high risk of becoming infected with MRSA. The longer they stay, the higher the risk. Patients with MRSA infections are less likely to survive than those with MSSA.

Cross Infection↗

Killing of cryptococcus neoformans by Staphylococcus aureus: the role of cryptococcal capsular polysaccharide in the fungal-bacteria interaction.

Microbes compete for the environmental niche which is their host. To investigate the effects of a pathogenic bacterium on invasion and colonization by a pathogenic yeast, Cryptococcus neoformans was co-cultured with Staphylococcus aureus. We found that the number of colony forming units of C. neoformans was decreased by Staphylococcus aureus. In contrast, the viability of Candida albicans was not affected. Under the microscope, wild-type C. neoformans cells were shown to be surrounded by S. aureus, while cells of a capsuleless mutant of C. neoformans were not. C. neoformans was not killed when a membrane separated it from S. aureus in co-culture. Killing was confirmed by staining with cyanoditolyl tetrazolium chloride: S. aureus stained red, indicating viability, while C. neojormans did not stain, indicating lethality. The in situ terminal deoxynucleotidyl transferase-mediated dUTR nick end labeling (TUNEL) assay indicated cell death with fragmentation of DNA of C. neoformans. Capsular polysaccharide from C. neoformans inhibited the killing. Treatment of the crude polysaccharide with protease increased the inhibition. The protective activity resided in the glucuronoxylomannan (GXM) fraction, although the concentration required for the inhibition was high. These results suggest that S. aureus kills C. neoformans by a process that involves attachment to the cryptococcal capsule.

Cryptococcosis↗

The delivery of benzyl penicillin to Staphylococcus aureus biofilms by use of liposomes.

The delivery of benzyl penicillin [penicillin G (pen-G)] encapsulated in cationic liposomes to a pen-G-sensitive strain of Staphylococcus aureus immobilized in biofilms has been investigated. The cationic liposomes prepared by extrusion (VETs, diameter approximately 140 nm) were composed of dipalmitoylphosphatidylcholine (DPPC), cholesterol, and dimethylammonium ethane carbamoyl cholesterol (DC-chol) at a molar ratio of 1.0:0.49:0.43. This composition containing 22 mole% of the cationic lipid DC-chol has been found previously (Kim et al. Colloids Surfaces A 1999, 149, 561-570) to be optimum for adsorption of the liposomes on S. aureus biofilms. The effectiveness of the liposomes to deliver pen-G to the biofilms immobilized on microtitre plates was assessed from the rate of growth of the cells after exposure to the liposomal drug carrier relative to free pen-G at the same concentration. The time to reach maximum growth rate from biofilms was investigated as a function of overall drug concentration in a range 2.9 x 10- 3 mM to 1.09 mM and as a function of time of exposure to liposomal drug in a range 1.5 s to 2 h. Liposomal drug delivery was most effective relative to free drug at low overall drug concentrations and short times of exposure. The time to reach maximum growth rate from S. aureus biofilms could be extended by a factor of approximately 4 relative to free drug by the use of liposomally encapsulated pen-G. The results were supported by direct measurements of the distribution of pen-G between biofilm and supernatant which showed enhanced values relative to free drug and a transient preferential uptake of drug induced by the liposomes. The study demonstrates that for low drug concentrations and short exposure times liposomal drug delivery greatly enhances the effectiveness of pen-G for inhibiting the growth of bacterial biofilms of the potentially pathogenic bacterium Staphylococcus aureus.

Adsorption↗