[EDTA disodium salt as a factor modifying Staphylococcus aureus sensitivity to penicillin. II. Effect on Staphylococcus aureus in its various growth phases].
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OBJECTIVES: Oral treatment regimens for multiresistant methicillin-resistant Staphylococcus aureus (MRSA) infections are limited. In Australia, rifampicin plus fusidic acid is the usual treatment regimen following glycopeptide therapy but many patients are intolerant of this; some isolates are resistant; new oxazolidinones are expensive for routine use. Pristinamycin is a possible alternative and we report our experience with this agent. METHODS: The Department of Microbiology and Infectious Diseases, South Western Area Pathology Service treats patients drawn from the South Western Sydney Area Health Service that houses approximately 800,000 people and contains approximately 2000 acute care public hospital beds. Patients prescribed pristinamycin between 1 September 2000 and 31 January 2000 were identified from hospital pharmacy records. A retrospective chart review was performed. Accepted clinical definitions of osteomyelitis and septic arthritis were used. RESULTS: Twenty-seven patients were identified with osteoarticular infections. Twenty-four cases involved Staphylococcus aureus (multiresistant MRSA in 21 cases); three involved Staphylococcus epidermidis sensu stricto; four cases involved multiple organisms. Nineteen cases received pristinamycin monotherapy; the others received various combinations (fusidic acid with five; other antibiotics with three). Therapy was generally well tolerated; no haematological or biochemical toxicity was detected. Seven patients had minor gastrointestinal disturbance; and one developed rash. Four patients required dose reduction. Only four patients ceased pristinamycin due to intolerance. Treatment outcome was evaluated in 23 cases; cure was effected in 16 cases, five were successfully suppressed and two failed. There were no deaths. CONCLUSIONS: Oral pristinamycin is well tolerated and an important additional agent to treat osteoarticular infections with multiresistant MRSA and other staphylococci.
Staphylococcus aureus is an important cause of pododermatitis, subcutaneous abscesses and mastitis in rabbits. On rabbitry level, two types of S. aureus infections can be distinguished. In the first type, caused by low virulence strains, the infection affects only a small number of animals. The second type of infection is caused by high virulence strains and spreads throughout the rabbit flock. The pathogenic capacity of a particular S. aureus strain is attributed to a combination of invasive properties and extracellular factors such as toxin production. Therefore, 22 high virulence and 37 low virulence S. aureus isolates were compared regarding the prevalence of genes encoding exfoliative toxins, leucotoxins and superantigens. This study revealed a clearly significant difference between HV and LV rabbit S. aureus strains. All typical HV isolates were positive for the egc cluster, containing the enterotoxin(like) genes seg, sei, selm, seln, selo and selu, whereas these genes were not detected in any of the LV isolates. Further research is necessary to clarify the importance of the egc cluster in the pathogenesis of infections with high virulence S. aureus strains in rabbits.
Staphylococcus aureus is a common pathogen responsible for nosocomial and community infections. It readily colonizes indwelling catheters, forming microbiotic communities termed biofilms. S. aureus bacteria in biofilms are protected from killing by antibiotics and the body's immune system. For years, one mechanism behind biofilm resistance to attack from the immune system's sentinel leukocytes has been conceptualized as a deficiency in the ability of the leukocytes to penetrate the biofilm. We demonstrate here that under conditions mimicking physiological shear, leukocytes attach, penetrate, and produce cytokines in response to maturing and fully matured S. aureus biofilm.
Staphylococcus aureus infections are often life threatening. Relatively little is known about the host response to these infections, in particular, the role played by cytokines. We established a mouse model of bacteremic S. aureus infection to correlate bacteriologic findings and pathologic changes with cytokine gene expression. Bacterial density in blood and tissue was highest at 1 h and minimal by 48 h. Despite the rapid clearance of bacteria, pathologic abnormalities and inflammatory cytokines were detected after clearance of the bacteria. The number of infiltrating inflammatory cells, as well as the size of inflammatory foci, increased with time. Interstitial accumulation of inflammatory cells and tissue damage, such as microabscesses, edema, and necrosis progressed following clearance of bacteria from the tissues. Levels of tumor necrosis factor and interleukin-1 protein in serum were detectable at 1 h and peaked at 4 h. Interleukin-6 protein expression showed different kinetics, with low levels detected at 1 h and increasing levels at 72 h postinfection. Tumor necrosis factor and the interleukins were expressed in inflammatory and noninflammatory cells in lung, liver, and heart tissues. Leukocytes in the infected tissues were highly reactive with antibodies to the three cytokines, suggesting that activated leukocytes are a major source of inflammatory cytokines after staphylococcal infection. Expression of interleukin-1 and interleukin-6 in tissue-specific cells and endothelial cells was also detected in infected tissues, indicating that cells other than leukocytes contribute to the elevated cytokine levels in this model. Once initiated, expression of inflammatory cytokines contributes to the pathogenesis of S. aureus disease.
Staphylococcus aureus has been shown to invade and induce the death of various cell types. Here we investigate whether the cytotoxicity of intracellular S. aureus is a general feature or rather characteristic of individual S. aureus strains. The majority of 23 randomly collected clinical S. aureus isolates were killed inside keratinocytes and fibroblasts, indicating that the uptake of S. aureus represents an important mechanism of cell-autonomous host defense. However, seven independent S. aureus isolates survived intracellularly and induced significant cytotoxicity for their host cells. Subcloning analysis revealed that the ability or inability to kill host cells is a stable, apparently genetically determined trait of a given S. aureus isolate. We show that noncytotoxic strains but not cytotoxic strains colocalize with the lysosomal marker LAMP-1, suggesting that only cytotoxic strains escape degradation by the endolysosomal pathway. In a mouse septicemic model, cytotoxic S. aureus isolates produce significantly greater lethality (96%) compared to noncytotoxic strains (41%), which corresponds to 23-, 63-, and 30,000-fold increases of bacterial loads in the liver, spleen, and kidney, respectively. Finally, cytotoxic S. aureus strains produce clinically apparent arthritis in mice at a greater frequency than compared to noncytotoxic S. aureus strains. The results of our study unravel a previously unrecognized dichotomy of cytotoxic and noncytotoxic S. aureus isolates, which may play an important role in the dissemination of, and mortality induced by, S. aureus infection.
Staphylococcus aureus is an important pathogen of humans, causing a range of superficial and potentially life-threatening diseases. Infection of the fruit fly Drosophila melanogaster with S. aureus results in systemic infection followed by death. Screening of defined S. aureus mutants for components important in pathogenesis identified perR and pheP, with fly death up to threefold slower after infection with the respective mutants compared to the wild-type. Infection of D. melanogaster with reporter gene fusion strains demonstrated the in vivo expression levels of the accessory gene regulator, agr, alpha-toxin, hla, and a manganese transporter, mntA. The use of the green fluorescent protein as a reporter under the control of the agr promoter (P3) showed S. aureus microcolony formation in vivo. The disease model also allowed the effect of antibiotic treatment on the flies to be determined. D. melanogaster is a genetically tractable model host for high-throughput analysis of S. aureus virulence determinants.
Staphylococcus aureus bacteraemia (SAB) is common. Around 8000 cases occur per year in Australia, of which 60% are hospital- or healthcare-associated. Risk factors for SAB include injectable drug use, haemodialysis, indwelling vascular catheters and immunosuppression. Metastatic infection develops in up to one-third of patients with SAB, with joints and heart valves being the most commonly affected sites. Community-acquisition,persistent fever, positive blood cultures after 48 h of treatment and the presence of embolic lesions correlate with the presence of complicated SAB (i.e. high risk of endocarditis and/or other metastatic complications). All patients require careful clinical evaluation to exclude endocarditis and other metastatic foci. Echocardiography,preferably transoesophageal echocardiography, should be performed to exclude endocarditis. Most patients with SAB, and all with features of complicated SAB, require prolonged intravenous antibiotic therapy (at least 4 weeks), but a subgroup with good prognostic features may be suitable for shorter intravenous therapy (2 weeks). Penicillinase-resistant penicillins (e.g.flucloxacillin) are the agents of choice for SAB with methicillin-sensitive strains. Vancomycin or first-generation cephalosporins are alternatives but have lower antimicrobial activity than flucloxacillin. However, vancomycin remains the therapy of choice for SAB due to methicillin-resistant strains. Combination therapy with gentamicin may be useful for the first few days of treatment in selected patients, but otherwise there are few data to support the use of combination regimens in SAB. Newer agents such as linezolid and quinupristin/dalfopristin may have a role in selected patients, especially in SAB due to S. aureus strains with reduced susceptibility to vancomycin.
Daptomycin is an investigational lipopeptide antibiotic active against gram-positive organisms. The mechanism of action is unique, resulting in interference with cell membrane transport. The bactericidal activity of daptomycin was evaluated against glycopeptide-intermediate susceptible Staphylococcus aureus (GISA), vancomycin-resistant Enterococcus faecium (VREF), and methicillin-resistant S. aureus (MRSA) in an in vitro infection model with simulated endocardial vegetations. Simulated regimens of daptomycin at 6 mg/kg/day (D6) and 10 mg/kg/day (D10) were utilized. MICs and MBCs for daptomycin were determined in the absence and in the presence of albumin with the following results (MIC/MBC): for GISA-992, 0.5/1.0 and 16/16; for VREF-590, 2.0/2.0 and 32/32; and for MRSA-494, 0.25/0.25 and 1.0/4.0 microg/ml, respectively. During the first 8 h daptomycin significantly reduced the inoculum for all organisms. Daptomycin at 6 mg/kg/day and 10 mg/kg/day had log(10) CFU/g reductions of 5 and 6, 3.4 and 5, and 6.4 and 6.5 by 8 h for GISA-992, VREF-590, and MRSA-494, respectively. Against both GISA-992 and VREF-590, the D10 regimen achieved the limit of detection at 72 h, with D6 regimens showing slight regrowth. A concentration-dependent killing effect was noted to occur, with daptomycin demonstrating a more rapid and greater kill from the D10 versus the D6 regimen. The results of this study suggest that daptomycin demonstrates significant (P < 0.05) activity against gram-positive organisms in a simulated sequestered infection site.
Staphylococcus aureus bacteraemia increases in frequency, and it is still a life-threatening disease. In recent years, some interesting studies such as the need for focus identification and the focus eradication have been performed. The aim of this review is to present an up-to-date assessment of the current challenges in the management of S. aureus bacteraemia in order to improve the outcome.
BACKGROUND: Staphylococcus aureus(S. aureus) is thought to play a significant role in the exacerbation of atopic dermatitis (AD). DS-Nh mice, a non-hair-bearing mouse model of AD, spontaneously develop dermatitis under conventional conditions. A remarkable increase in S. aureus is considered to strongly relate to the induction and aggravation of this dermatitis. A topical use of anatase-type titanium dioxide (TiO2) followed by UV irradiation, acting as photocatalyst, is believed to have antibacterial activity. We investigated the bactericidal effect of TiO2 with UV irradiation on DS-Nh mice to prevent the aggravation of the dermatitis. METHODS: Ten-week-old DS-Nh mice were treated with TiO2 in petrolatum on the back, followed by UVA irradiation, for 11 weeks. The severity of dermatitis was assessed by evaluating individual lesions using a 4-grade scale and expressed as the total skin score. S. aureus colonizing the mouse skin was counted after isolation and incubation with agar medium. The skin barrier dysfunction was evaluated by measuring transepidermal water loss (TEWL). RESULTS: The mice treated with TiO2 and UV irradiation showed a significant increase in total skin scores, the number of S. aureus and TEWL values, compared with non-treated mice. In contrast, these parameters were significantly lower in the mice treated with petrolatum and UV irradiation. CONCLUSIONS: A significant increase in S. aureus was recognized on the skin together with the aggravation of AD-like dermatitis in our mice model. Skin barrier dysfunction induced by TiO2 and UV irradiation seems to facilitate the increase in S. aureus.
OBJECTIVE: To determine the comparative virulence of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S aureus (MSSA) by consideration of predisposing factors and outcomes in patients infected with these organisms in the healthcare setting. DESIGN: Analysis of an historical cohort of 504 bacteremic patients (316 MSSA and 188 MRSA), examining factors associated with mortality. SETTING: A 916-bed, university-affiliated, tertiary referral hospital. RESULTS: Risk factors for the development of MRSA include male gender, admission due to trauma, immunosuppression, presence of a central vascular line or an indwelling urinary catheter, and a past history of MRSA infection. Overall mortality was 22%. Death due to bacteremia was significantly greater in the MRSA group (risk ratio, 1.68; P<.05), although these patients were not found to be more likely to die due to underlying disease during treatment of bacteremia. In those patients who recovered from bacteremia, no significant differences for the outcome of death could be determined between the MRSA and MSSA groups. CONCLUSIONS: There is a general consensus in the published literature that MRSA bacteremia is more likely to be associated with death, and we confirm this conclusion. However, in contrast to other studies, our MRSA cohort does not appear to be more at risk of death due to underlying disease during treatment for bacteremia. Similarly, the general consensus that MRSA patients have an increased overall mortality was not confirmed in our study. Differences in comorbidities of patients may provide some explanation of these conflicting results, while an alternate explanation is that MRSA strains are more virulent than MSSA in some centers. Perhaps the most plausible explanation is that treatment is provided earlier and in a more aggressive fashion in some centers, leading to an overall lower mortality rate in all staphylococcal bacteremias in these institutions.
Staphylococcus aureus is the most common bacterial species found in nongonococcal bacterial arthritis in humans. We present the first description, to our knowledge, of an outbreak of spontaneous staphylococcal arthritis in a rat colony. In a group of 10 rats, 9 displayed arthritis. Clinically, the most obvious findings were arthritis of one or both hindpaws and malaise. Bacteriophage typing showed the common phage type 85 in isolates recovered from the joints, blood, and bedding of rats and from the nose and cheeks of one person from the staff of the animal facility. The S. aureus strain proved to produce staphylococcal enterotoxin A and exhibited strong binding to collagen types I and II and bone sialoprotein, which are potentially important virulence factors. When the recovered S. aureus strain was injected intravenously into healthy rats, severe septic arthritis was induced in almost all of the animals. The arthritic lesions were characterized by infiltration of phagocytic cells and T lymphocytes into the synovium. Many of the synovial cells strongly expressed major histocompatibility complex class II molecules. Increased levels of interleukin 6 in serum as well as a prominent polyclonal B-cell activation were noted throughout the disease course. Pretreatment of S. aureus-injected rats in vivo with an antibody to the alpha beta T-cell receptor significantly decreased the severity of the arthritis. Our results indicate that alpha beta + T lymphocytes contribute to an erosive and persistent course of S. aureus arthritis.
BACKGROUND: Staphylococcus aureus causes a multiplicity of human diseases acquired in community and healthcare settings alike around the globe. While most studies focus on coding changes to assess genome evolution and study genetic adaptation, interrogation of silent mutations in the form of synonymous codon usage bias is less well-studied. As such, understanding of patterns in codon bias at the gene and genome levels, and how codon bias impacts protein expression in S. aureus remains incomplete. METHODS: The codon bias of 2,565 protein encoding genes from NCTC 8325 was queried against all publicly available closed S. aureus genomes. Using public BioSample data, genomes were sorted by disease state, submitting institution, and collection site. Codon bias was assessed at the level of gene and genome using the codon adaptation index (CAI), calculated using 30S and 50S ribosomal genes. Gene set enrichment analysis was applied to determine associations between physiological functions, CAI gene scores, and interquartile ranges. CAI scores were also compared to an in vitro S. aureus proteomics database to correlate codon bias and protein expression. RESULTS: CAI scores varied within and between isolates at the gene and genome levels. Genes with ribosome-associated functions were most enriched among high CAI genes, and had low CAI interquartile ranges (IQR), suggesting selective pressure to maintain high expression of these genes across all S. aureus isolates. Genome sequences submitted by Aga Khan University Hospital, Nairobi, Kenya were most different from others. For the LAC USA 300 strain, CAI and protein expression were moderately positively correlated (cor = 0.534, p < 2.2e-16). CONCLUSIONS: Codon bias in S. aureus was shown to vary between gene, and to be a source of genetic variation between isolates; CAI and in vitro protein expression were positively correlated.
Day-old chicks were subcutaneously infected with Staphylococcus aureus strains of site variety gallinae and crystalviolet Type A. The lysis patterns involved were with phages 84 as well as with phages 53, 77, and 84 of the international base scheme for human staphylococci. In fowl with synovitis and arthritis, osteomyelitis, dermatitis, and septicaemia, 52 of 57 tested strains (91.2 per cent) produced on the site of injection pathognomic oedematous-haemorrhagic and necrotising changes of the skin and hypodermis (haemorrhagic dermatitis) in a way which is characteristic also of natural diseases. However, no specific skin changes could be produced by subcutaneous infection of nine strains of site variety hominis or undefinable category with differentiated biochemical properties and lysis patterns. They had been isolated from cases of compound infection, salpingitis and wound infections as well as from one case of arthritis. Fifty-seven strains, obtained from mammals, were tested subcutaneously, on day-old chicks, among them 21 strains of site varieties of bovis, canis, ovis, and hominis. However, only four of those strains, isolated from miscarried pig foetuses and cattle with mastitis, caused, oedematisation and exudation (7.02 per cent), but no haemorrhagia and only minor lysis and necrosis of skin. The minimum dose required to induce haemolytic dermatitis was 11 x 10(3) pathogens, and the lowest lethal dose of six tested strains was between 11 x 10(5) and 26 x 10(7) pathogens. The lowest dose of mammalian strains with lethality to day-old chicks was [10 bis]10(4) times as high as the lowest dose of fowl strains. Subcutaneous infection of day-old chicks has worked well as a high-speed test by which to differentiate between fowl-pathogenic Staphylococcus aureus strains, on the one hand, and mammalian strains, on the other. The need for delimitation of such strains has been confirmed by the above experimental infections. Against the background of the above results, guidelines are proposed for the control of Staphylococcus aureus infection of fowl.
Staphylococcus aureus S-6 cells grown in chemically defined media often lysed after exponential growth. Lysis could be prevented by the addition of alanine or proline before the culture reached stationary phase.
PURPOSE: Staphylococcus aureus is a common cause of ocular infection and inflammation. We hypothesized that potential for S. aureus to cause an ocular infection would be enhanced if these bacteria are able to adhere to the biomaterials used in contact lenses. In turn, bacterial adhesion could also be influenced by other factors, such as properties of contact lenses and the absorbance of some tear components. We investigated the effect of the tear protein lysozyme on S. aureus adhesion to contact lenses and its effect on production of toxins or enzymes. METHODS: Bacterial adhesion on contact lenses was determined by counting the total number of bacteria as well as viable bacteria on lysozyme-coated or non-coated lenses, and by counting bacteria grown in the presence or absence of lysozyme in the medium. Toxin and enzyme production was assessed by haemolysis and proteolysis assays. RESULTS: Our results indicate that adhesion was significantly increased in the presence of lysozyme, both in the medium and coated onto contact lenses (P=0.04). The presence of lysozyme did not alter the production of alpha- or beta- toxins. However, the presence of lysozyme inhibited elastase activity. CONCLUSION: These results indicate that lysozyme deposition on contact lenses promoted S. aureus adhesion. The tear protein lysozyme might modify elastase activity and thus modulate the production of corneal degradation resulting from the action of this enzyme.
In Staphylococcus aureus, enterotoxin B (SEB) is a superantigen that activates host interleukins and induces adverse responses, ranging from food poisoning to toxic shock. The alternate sigma factor, sigmaB (SigmaB), and agr are two known regulators of S. aureus. Northern blots of strain COL, a sigB-positive strain, showed an inverse correlation between sigmaB expression and seb message. seb expression was also measured as a function of a seb promoter linked to green fluorescent protein (GFP) expression in RN6390, COL, and Newman. In sigB mutants of RN6390, SH1000, COL, and Newman, seb promoter activities, as measured by GFP expression, increased relative to the respective parental types but at differing levels, suggesting alternate strain-specific regulation. In agr mutants of RN6390 and Newman, seb promoter activities were intermediate between the high level seen for the sigB mutant and the low level in the sigB active strains. A sigB agr double mutant of RN6390 displayed lower GFP expression than the agr mutant. These results suggest that while sigmaB and agr regulate seb expression in a divergent manner, other activator(s) of seb that depend on sigB expression may be present in S. aureus.