Analysis of genomic diversity within the Xr-region of the protein A gene in clinical isolates of Staphylococcus aureus. (49/7250)

Protein A of Staphylococcus aureus contains a polymorphic Xr-region characterized by a tandem repeat of eight amino acid units. In this study, the diversity of genes encoding the repeat regions and their relatedness among S. aureus strains was analyzed. Ten different protein-A types characterized by repeat numbers 4-13 were identified in a total of 293 clinical isolates. The protein-A type with 10 repeat units (10 repeats) in the Xr-region was most frequently detected in methicillin-resistant S. aureus, whereas the majority of methicillin-susceptible strains were distributed almost evenly into protein-A types with 7-11 repeats. Strains that belonged to a single coagulase type were classified into multiple protein-A types, e.g. strains with the common coagulase types II and VII were differentiated into 7 and 8 protein-A types, respectively. Nucleotide sequence analysis of the Xr-region of 42 representative strains revealed the presence of 37 different genotypes (spa types), which were constituted by a combination of several of 24 different repeat unit genotypes. Based on the similarity in arrangement of repeat unit genotypes, 34 strains with different repeat numbers were classified into 5 genetic clusters (C1-C5). The clusters C1, C2 and C3 consisted exclusively of strains with identical coagulase types II, III, and IV, respectively. These findings suggested that the protein-A gene of S. aureus has evolved from a common ancestral clone in individual clusters independently.  (+info)

Coagulase gene polymorphism of Staphylococcus aureus isolates from dairy cattle in different geographical areas. (50/7250)

The objectives of this study were to investigate the coagulase gene polymorphism of Staphylococcus aureus isolates obtained from bovine mastitic milk and to determine the resistance of predominant and rare coagulase genotypes to bovine blood neutrophil bactericidal activities. A total of 453 isolates were collected from four countries: the Czech Republic, France, Korea and the United States. The isolates were subtyped into 40 types by restriction fragment length polymorphism (RFLP) of the coagulase gene. Twenty-three strains from predominant and rare genotypes were evaluated for their ability to resist neutrophil bactericidal activities. There were significant (P < 0.01) differences in the average percent neutrophil killing of the predominant (16.7%) and rare (39.7%) genotypes when bacteria were opsonized with antiserum. The results indicate that the profiles of coagulase genotype differ among geographic locations, and only a few genotypes prevail in each location. In addition, the predominant genotypes were more resistant to neutrophil bactericidal activities than rare genotypes.  (+info)

Protection by group II phospholipase A2 against Staphylococcus aureus. (51/7250)

Group II phospholipase A2 (PLA2) is an enzyme that has marked antibacterial properties in vitro. To define the role of group II PLA2 in the defense against Staphylococcus aureus, we studied host responses in transgenic mice expressing human group II PLA2 and group II PLA2-deficient C57BL/6J mice in experimental S. aureus infection. After the administration of S. aureus, the transgenic mice showed increased expression of group II PLA2 mRNA in the liver and increased concentration of group II PLA2 in serum, whereas the PLA2-deficient mice completely lacked the PLA2 response. Expression of human group II PLA2 resulted in reduced mortality and improved the resistance of the mice by killing the bacteria as indicated by low numbers of live bacteria in their tissues. Human group II PLA2 was responsible for the bactericidal activity of transgenic mouse serum. These results suggest a possible role for group II PLA2 in the innate immunity against S. aureus infection.  (+info)

Bacterial conjunctivitis in Muc1 null mice. (52/7250)

PURPOSE: In contrast to wild-type mice, genetically engineered Mucin1 (Muc1) null animals display a marked propensity for development of blepharitis and conjunctivitis. Molecular approaches confirmed the presence of Muc1 mRNA and protein in the conjunctival tissue of wild-type mice and identified the bacterial species in Muc1 null symptomatic mice. METHODS: Muc1 null animals housed in a conventional facility were examined for visually apparent inflammation of the eye and surrounding tissue. Blood taken from overtly affected animals was assayed for antibodies to common murine viral agents. Swabs of infected eyes and whole eye preparations were used to detect and speciate bacterial pathogens. Frozen sections of whole eye, lid margin, and Harderian gland were immunostained with antibodies to Muc1 and cytokeratin 14, both epithelial cell markers. Northern blot analysis and reverse transcription-polymerase chain reaction (RT-PCR) were performed on RNA isolated from conjunctiva and Harderian gland of wild-type mice to compare relative levels of transcript. RESULTS: Student's unpaired t-test performed on the eye inflammation frequency of Muc1 null mice confirmed a statistical significance (P < 0.01) when compared to wild-type background animals housed in the same room. Analysis of blood samples from affected Muc1 null animals detected no common murine viral pathogens. Bacterial analysis of conjunctival swabs and whole eye preparations demonstrated the presence of coagulase-negative Staphylococcus, Streptococcus type alpha, and Corynebacterium group G2. Muc1 antibody staining of wild-type sections revealed the presence of Muc1 on conjunctival goblet and non-goblet cells and on the epithelium of the Harderian gland. Serial sections stained with cytokeratin 14 antibody confirmed the epithelial nature of cells expressing the Muc1 protein. RNA from conjunctiva and Harderian gland subjected to RT-PCR and northern blot analysis showed an abundance of Muc1 transcript in these tissues. CONCLUSIONS: Muc1 mRNA and protein are present in murine conjunctival and Harderian gland epithelia. Animals lacking Muc1 mRNA and protein are predisposed to developing eye inflammation when compared to wild-type animals with an intact Muc1 gene. Muc1 appears to play a critical protective role at the ocular surface, presumably by acting as a barrier to infection by certain bacterial strains.  (+info)

Bacterial resistance of refrigerated and cryopreserved aortic allografts in an experimental virulent infection model. (53/7250)

PURPOSE: The bacterial resistance of refrigerated and cryopreserved aortic allografts in a highly virulent infection in a dog model was studied. METHODS: The infrarenal aorta of 12 dogs was replaced with either a cryopreserved aortic allograft (group I, n = 6) or a refrigerated aortic allograft (group II, n = 6) in infected sites. Allografts were harvested from dogs and stored for 1 week, either by cryopreservation (-140 degrees C) or refrigerated method (4 degrees C), in a preservation medium. At the time of implantation, induction of infection was achieved with an infected piece of knitted Dacron placed just beneath the allograft. The Dacron was contaminated in vitro by soaking it in a solution with Staphylococcus aureus PR209. All 12 dogs received no adjunct antibiotic or antithrombotic therapy. Four weeks after implantation, the animals were killed to recover the grafts for bacteriological and histological analyses. Bacterial results were expressed as colony-forming units (CFU)/cm2 of graft material. RESULTS: In group I, only one allograft grew bacteria at 2. 16 x 10(6 )CFU/cm2, with a blood culture positive for S aureus. In group II, one dog died at 3 weeks from a false septic aneurysm rupture, all the allografts were infected (P <.05) with a mean bacterial count of 9.41 +/- 6.8 x 10(4) CFU/cm2, and three blood cultures were positive for S aureus. The patency of the grafts was analyzed at the time of recovery. Three laminar thrombi without occlusion were present in group I; none were present in group II. A better preserved endothelium in group I was revealed by means of histologic analysis staining with factor VIII antibody before implantation. After 4 weeks of implantation in the infected site, infected allografts presented polynuclear infiltrates in the media with a high degree of inflammatory reaction, and endothelial recovery was more significant in group I, with numerous young plump cells. CONCLUSION: This study demonstrates that cryopreserved allografts implanted in infected sites in a dog model can produce greater bacterial resistance.  (+info)

Clinical features and outcome of septic arthritis in a single UK Health District 1982-1991. (54/7250)

AIMS: To determine the clinical features of a large number of unselected UK hospital patients with confirmed septic arthritis and to determine those features associated with a poor outcome. STUDY DESIGN: Retrospective, case-note survey. SETTING: A single English Health District. PATIENTS: All patients admitted to hospital in Nottingham during the period 1 January 1982 to 31 December 1991 with confirmed septic arthritis were included. OUTCOME MEASURES: Death, osteomyelitis and recorded functional impairment. RESULTS: The spectrum of causative organisms remains similar to that seen in previous studies with the Gram positive organisms Staphylococcus aureus and Streptococci responsible for 74% of cases, gonococcal infections though were less common. Culture of joint aspirates and or blood were positive in 82% of cases, with the Gram stain demonstrating the causative organism in 50% of cases. Pre-existing joint disease was evident in 35% of cases. The mortality remains high at 11.5% with a significant additional morbidity of 31.6%. Multivariate analysis suggests that important predictors of death are: confusion at presentation, age > or = 65 years, multiple joint sepsis or involvement of the elbow joint, and of morbidity are: age > or = 65 years, diabetes mellitus, open surgical drainage, and Gram positive infections other than S aureus. CONCLUSIONS: Septic arthritis continues to be associated with a considerable degree of morbidity and mortality. These results confirm the importance of obtaining synovial fluid and blood for culture before starting antimicrobial treatment. The apparent poorer outcome found with surgical intervention is in line with some previous suggestions but should be interpreted with caution in light of the retrospective nature of this study.  (+info)

Studies of antibiotic resistance within the patient, hospitals and the community using simple mathematical models. (55/7250)

The emergence of antibiotic resistance in a wide variety of important pathogens of humans presents a worldwide threat to public health. This paper describes recent work on the use of mathematical models of the emergence and spread of resistance bacteria, on scales ranging from within the patient, in hospitals and within communities of people. Model development starts within the treated patient, and pharmacokinetic and pharmacodynamic principles are melded within a framework that mirrors the interaction between bacterial population growth, drug treatment and the immunological responses targeted at the pathogen. The model helps identify areas in which more precise information is needed, particularly in the context of how drugs influence pathogen birth and death rates (pharmacodynamics). The next area addressed is the spread of multiply drug-resistant bacteria in hospital settings. Models of the transmission dynamics of the pathogen provide a framework for assessing the relative merits of different forms of intervention, and provide criteria for control or eradication. The model is applied to the spread of vancomycin-resistant enterococci in an intensive care setting. This model framework is generalized to consider the spread of resistant organisms between hospitals. The model framework allows for heterogeneity in hospital size and highlights the importance of large hospitals in the maintenance of resistant organisms within a defined country. The spread of methicillin resistant Staphylococcus aureus (MRSA) in England and Wales provides a template for model construction and analysis. The final section addresses the emergence and spread of resistant organisms in communities of people and the dependence on the intensity of selection as measured by the volume or rate of drug use. Model output is fitted to data for Finland and Iceland and conclusions drawn concerning the key factors determining the rate of spread and decay once drug pressure is relaxed.  (+info)

Isolation and characterization of a sigB deletion mutant of Staphylococcus aureus. (56/7250)

The sigB gene of Staphylococcus aureus, coding for the alternate sigma factor B, has been deleted by allelic replacement mutagenesis. The mutant grew as well as the parent in vitro, although it was deficient in clumping factor, coagulase, and pigment. In two murine and one rat infection model the mutant showed no reduction in virulence.  (+info)