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DNA sequence-based detection of group B Streptococcus directly from heart valve tissue in a patient with culture-negative endocarditis.

A human case of culture-negative bacterial endocarditis is presented where conventional methods failed to determine a microbial aetiology. DNA sequencing performed directly on autopsy heart valve tissue revealed Streptococcus agalactiae (group B streptococcus). To our knowledge, this is the first report of DNA sequence-based detection of this organism directly from a heart valve.

Anti-Bacterial Agents↗

Non-instrumental immunoassay based on coloured polyacrolein latex: application to group-specific polysaccharide of Streptococcus pyogenes.

Non-instrumental immunoassay methods based on immunofiltration and microtiter particle agglutination (MPA) techniques have been developed using coloured polyacrolein latex. These methods have been applied to the quantification of the group-specific polysaccharide, A-PS, of S.pyogenes (group A Streptococcus) and compared to the standard ELISA tests. The assay with the ability to detect the lowest concentration of antigen was MPA; as little as 0.05 ng A-PS/ml or 10(4) cells/ml could be detected in 1.5 h. In comparison to ELISA test the sensitivity of MPA was 10 times higher and the procedure of the assay was much more simple. The sensitivity of the immunofiltration assay using both enzyme and latex markers was shown to be the same (50 ng A-PS/ml) and the duration of the assay 3-5 min. No cross-reactions of latex conjugates with non A Streptococcus cell lysates have been observed. The developed methods are easy to perform and require neither sophisticated equipment nor specially trained personal.

Acrolein↗

An immunological relationship between the group. A streptococcus and mammalian muscle.

By means of the immunofluorescent staining technique, antisera to a wide variety of serological types of Group A and A variant streptococci were found to contain an antibody which reacted with mammalian striated muscle, both skeletal and cardiac, as well as with smooth muscle in the endocardium and in the media of arterioles. Similar heart-reactive antibodies were not present in antisera to most other groups of hemolytic streptococci and to other Gram-positive cocci. Chemical and serological studies clearly pointed to the cell (protoplast) membrane of the Group A streptococcus as the locus of the antigenic determinant of this heart-reactive antibody. In addition, preliminary studies suggested that the reaction between this streptococcal antibody and cardiac tissue represented an immunological relationship between the sarcolemma, the membrane of a mammalian muscle cell and the cell membrane of a bacterium, the hemolytic streptococcus.

Antibodies↗

Characterization of the galU gene of Streptococcus pneumoniae encoding a uridine diphosphoglucose pyrophosphorylase: a gene essential for capsular polysaccharide biosynthesis.

The galU gene of Streptococcus pneumoniae has been cloned and sequenced. Escherichia coli cells harboring the recombinant plasmid pMMG2 (galU) overproduced a protein that has been shown to correspond to a uridine 5'-triphosphate:glucose-1-phosphate uridylyltransferase (uridine diphosphoglucose [UDP-Glc] pyrophosphorylase) responsible for the synthesis of UDP-Glc, a key compound in the biosynthesis of polysaccharides. A gene very similar to the S. pneumoniae galU has been found in a partial nucleotide sequence of the Streptococcus pyogenes genome. Knockout galU mutants of type 1 pneumococci are unable to synthesize a detectable capsule. An identical result was found in type 3 S. pneumoniae cells in spite of the fact that these bacteria contain a type-specific gene (cap3C) that also encodes a UDP-Glc pyrophosphorylase. Since eukaryotic UDP-Glc pyrophosphorylases appear to be completely unrelated to their prokaryotic counterparts, we postulate that GalU may be an appropriate target for the search of new drugs to control the pathogenicity of bacteria like pneumococcus and S. pyogenes.

Amino Acid Sequence↗

Group B Streptococcus colonization in male and nonpregnant female university students: a cross-sectional prevalence study.

We describe the prevalence of colonization with group B Streptococcus species in a random sample of otherwise healthy male and nonpregnant female college students. Colonization with group B Streptococcus species occurs at a high frequency among healthy students, and there was a suggestion that it is associated with having engaged in sexual activity, tampon use, milk consumption, and hand washing done < or =4 times per day. However, larger studies are needed to verify these findings.

Adolescent↗

Intrinsic reduced susceptibility of serotype 6 Streptococcus pyogenes to fluoroquinolone antibiotics.

BACKGROUND: Fluoroquinolone resistance is common in Staphylococcus aureus, is increasing in Streptococcus pneumoniae, and is reported in Streptococcus pyogenes. METHODS: We surveyed 384 clinical isolates of S. pyogenes, isolated during 2002-2003, for susceptibility to ciprofloxacin. We performed nucleotide sequencing of the parC and gyrA genes and determined the M/emm type for selected isolates. Additionally, we analyzed M/emm type 6 S. pyogenes isolated during 1918-2003 from diverse locations. RESULTS: Of the survey isolates, 10.9% had reduced zones of inhibition to ciprofloxacin in the disk-diffusion test and had elevated minimum inhibitory concentrations to other fluoroquinolones, compared with those of fully susceptible isolates. Of the resistant isolates, 90.5% were M/emm type 6, and all sequenced M/emm type 6 isolates contained a serine-to-alanine substitution at position 79 in parC. Strikingly, the same findings were also present in macrolide-resistant isolates from a recent outbreak of S. pyogenes infection in Pittsburgh and in the Lancefield reference strain of M type 6, which was isolated in 1918, decades before the development of fluoroquinolone antibiotics. CONCLUSION: M/emm type 6 S. pyogenes has intrinsic reduced susceptibility to fluoroquinolones, as a result of a polymorphism in parC. This finding was also demonstrated in erythromycin-resistant M/emm type 6 S. pyogenes, which raises concern for the emergence of multidrug-resistant S. pyogenes.

Adult↗

Nitric oxide is a key determinant of group B streptococcus-induced murine macrophage apoptosis.

Group B streptococcus (GBS; Streptococcus agalactiae) induces apoptosis of macrophages, and this may be an important mechanism GBS uses to suppress immune responses. The mechanisms whereby GBS induces apoptosis have not been identified. We studied GBS infection in murine macrophage-like J774A.1 cells and analyzed gene expression before apoptosis. Tumor necrosis factor (TNF)-alpha , interleukin (IL)-1, and inducible nitric oxide synthase (iNOS) gene expression coincided with apoptosis. Inhibition of iNOS gene expression by use of N(G)-monomethyl-L-arginine (NMMA) inhibited apoptosis, whereas inhibition of TNF-alpha and IL-1 biological activity did not. Macrophages from congenic iNOS-deficient mice were less susceptible to apoptosis than were macrophages from C57BL/6 mice. The NO donor S-nitroso-N-acetylpenicillamine (SNAP) induced apoptosis without infection, confirming its proapoptotic effect. NMMA did not impair the microbicidal activity of macrophages, however, and SNAP was not bactericidal against GBS in vitro. In human monocyte-derived macrophages (HMDMs), NO production was minimal, even after costimulation with IFN-gamma and lipopolysaccharide. Dose-dependent apoptosis of HMDMs occurred without a significant NO response. Thus, NO is an important mediator of GBS-induced murine macrophage apoptosis but does not contribute to antimicrobial activity or cytotoxicity in HMDMs. HMDMs and murine macrophages are killed by GBS by alternative, NO-independent mechanisms. Future studies of host-cell machinery commandeered by GBS to bring about apoptosis will be important for understanding the role played by apoptosis in defense against this important human pathogen.

Animals↗

Autolysis and cell wall degradation in a choline-independent strain of Streptococcus pneumoniae.

Streptococcus pneumoniae has an auxotrophic requirement for choline, and choline residues that incorporate into the wall and membrane teichoic acids are intimately involved with the control of autolytic phenomena of this bacterium. We report here the re-examination of the role of choline in autolytic cell wall degradation using the choline-independent S. pneumoniae strain R6Cho- recovered from a heterologous cross with DNA from Streptococcus oralis. S pneumoniae Cho- cultured in choline-free medium grew with normal generation time but formed long chains, failed to undergo stationary-phase autolysis, and was also resistant to lysis induced by deoxycholate or penicillin. Cell walls produced under these conditions had reduced phosphorus content, contained no choline residues detectable by nuclear magnetic resonance, and had reduced binding capacity for the pneumococcal autolytic amidase, and complete hydrolysis of such walls by the amidase required prolonged incubation with high concentrations of the enzyme. Addition of choline to the growth medium reversed at these phenomena. High-performance liquid chromatography analysis of amidase digests of cell walls prepared from strain R6Cho- grown with or without choline produced identical stem peptide profiles, which were also similar to that of the parental S. pneumoniae strain R6. Peptidoglycans prepared by hydrofluoric extraction of cell walls from Cho- growth with or without choline or from the parental strain R6 were uniformly susceptible to the autolytic amidase and were fully degraded to the normal family of stem peptides, indicating that, in sharp contrast to the case of cell walls, the amidase degradation of teichoic acid-free peptidoglycan did not require the presence of choline residues in the substrate.

Amidohydrolases↗

Penicillin-resistant Streptococcus pneumoniae in Colombia: presence of international epidemic clones. Colombian pneumococcal study group.

The global spread of multidrug-resistant Streptococcus pneumoniae clones is well documented in the literature. A study to determine type distribution and antimicrobial susceptibility of invasive pneumococcal isolates from Colombian children under the age of 5 was conducted from 1994 to 1996. Health centers in Santa Fe de Bogota, Medellin, Cali, and other cities collected 409 Streptococcus pneumoniae isolates. Diminished susceptibility to penicillin (DSP) was 15.6%; from these, 11.5% showed intermediate-level resistance (ILR) and 4.1% showed high-level resistance (HLR). Fifty-nine of the DSP isolates were examined by pulse field gel electrophoresis (PFGE). Capsular isolate types were 23F (54%), 14 (24%), 19F (10%), 6B (7%), 9V (3%), and 34 (2%). PFGE analysis revealed that 8 isolates shared the Spanish/USA international clone's characteristic features: PFGE pattern type A, serotype 23F; 87.5% exhibited HLR for penicillin, and all were resistant to trimethoprim/sulfamethoxazole (TMP-SMX), tetracycline, and chloramphenicol. Another 7 isolates showed the French/Spanish international clone's features: PFGE pattern type B, 2 of them being serotype 9V; and 5 type 14; HLR to penicillin was 71%, and all proved resistant to TMP-SMX. A large cluster of 24 isolates (41% of all isolates examined) shared a common PFGE type C, with 14 subtypes; all but one, serotype 34, were serotype 23F and had ILR to penicillin; 58% were resistant to TMP-SMX and 50% to tetracycline, but none presented erythromycin or chloramphenicol resistance. The remaining 20 isolates could be grouped into 12 different PFGE types; ILR was shown in 75% of isolates, 70% were resistant to TMP-SMX and to tetracycline, 15% were resistant to erythromycin, and none were resistant to chloramphenicol. These data suggest that some Colombian isolates are clonally related to two of the well-known international epidemic S. pneumoniae clones.

Colombia↗

Polyclonal spread of erythromycin-resistant Streptococcus agalactiae in southern Taiwan.

Resistance to erythromycin is common among Streptococcus agalactiae in Taiwan, however the genetic relatedness of erythromycin-resistant isolates has not yet been reported. From 1991 to 2001, 629 clinical isolates of S. agalactiae were collected in a medical center at Tainan in southern Taiwan, of which 189 (30.0%) were resistant to erythromycin. The isolation rate of erythromycin-resistant group B streptococcus (GBS) was stable, irrespective of the clinical sources or study period. Among them, 145 (76.7%) isolates showed the macrolide-lincosamide-streptogramin B (MLS)-resistant phenotype, and 44 (23.3%) had the macrolide (M)- resistant phenotype. Of the isolates with MLS phenotype, 141 (97.2%) isolates harbored the ermB gene alone and only three (2.1%) the ermTR gene, whereas 41 (93.2%) of 44 isolates with M phenotype harbored the mefA/E gene. Of 177 typeable isolates, there were 26 unrelated pulsed-field gel electrophoresis (PFGE) patterns. PFGE type 1 accounted for 17.8% (24/135) of MLS phenotype isolates with the ermB gene and 48.7% (18/37) of M phenotype isolates with the mefA/E gene. During the study period, the proportion of PFGE type 6 decreased significantly, whereas that of type 8 increased. Our results suggest that erythromycin resistance is not uncommon among clinical isolates of S. agalactiae and is, at least, partially related to polyclonal spread in southern Taiwan.

Drug Resistance, Bacterial↗

Meningitis caused by Streptococcus dysgalactiae in a preterm infant.

This report describes a case of meningitis caused by a Lancefield group C streptococcus (Streptococcus dysgalactiae) in a 9-week-old infant. Bacteria of this group rarely cause serious infections in man. The organism was identified as a member of Lancefield group C by the acid extraction method and as S. dysgalactiae by biochemical tests. The patient's condition responded well to penicillin and tobramycin therapy, with no obvious neurologic sequelae.

Anti-Bacterial Agents↗

Genetic regulation of host responses to group A streptococcus in mice.

The group A streptococci (GAS, Streptococcus pyogenes) are important human pathogens which can cause a variety of diseases, ranging from mild infections to very severe invasive diseases. In recent years, evidence has been accumulated that host genetic factors have a major influence on the outcome of streptococcal infections. Variability in the degree of susceptibility of different inbred mouse strains to infection with GAS has demonstrated that the host genetic background largely determines the susceptibility of mice to this pathogen. This information is particularly useful for studying the immune mechanisms underlying disease susceptibility in mice, and provides an entry point for the identification of host defence loci. This paper reviews the recent advances in the characterisation of pathogenic mechanisms associated with the development of GAS-induced septic shock in the mouse model and outlines the current knowledge regarding the genetic control of immune responses to Group A streptococcus in mice.

Animals↗

Streptococcus salivarius bacteremia and meningitis following upper gastrointestinal endoscopy and cauterization for gastric bleeding.

Streptococcus salivarius is a normal inhabitant of the oral cavity. When isolated from blood cultures, this viridans streptococcus is often disregarded as a contaminant. Viridans streptococci, although a common cause of endocarditis, are rarely associated with bacterial meningitis and account for less than 1% of all cases of purulent meningitis. We report a case of bacteremia and meningitis due to S. salivarius that occurred in a patient who underwent upper gastrointestinal endoscopy and cauterization for control of gastric bleeding. Although bacteremia following gastrointestinal procedures has been well documented, its incidence is low (ranging from 2% to 10%), and its course is usually transient without major clinical sequelae. This case is unique with respect to the pathogen isolated, the length of bacteremia, and the meningitis that subsequently developed following a gastroesophageal procedure.

Bacteremia↗

Antimicrobial resistance in Streptococcus pneumoniae: a South African perspective.

Resistance to penicillin among South African strains of Streptococcus pneumoniae increased from 4.9% in 1979 to 14.4% in 1990. Except for resistance to co-trimoxazole (44%), resistance to other antimicrobial agents remained relatively low. Multiply resistant strains belonged mainly to serovars 6B, 19A, 14, and, more recently, 23F. Use of chloramphenicol to treat meningitis caused by strains relatively resistant to penicillin proved to be unsatisfactory, probably because of the inadequate bactericidal activity of chloramphenicol against these strains. Spread of penicillin-resistant nasopharyngeal strains in pediatric wards was most common among children who received antimicrobial therapy. Penicillin-binding protein (PBP) patterns were shown to vary in resistant clinical strains. Interspecies transfer of penicillin resistance between Streptococcus mitis and S. pneumoniae was demonstrated and antigenic homology was found in PBPs 1A and 2B of strains belonging to these species. Restriction enzyme mapping following DNA amplification of the PBP 2B gene revealed six arrangements among South African strains within serogroup 19. Despite extensive studies in South Africa and several other countries, many questions with regard to the global problem of antimicrobial resistance among S. pneumoniae strains remain unanswered, especially those that relate to prevalence in developing regions of the world.

Carrier State↗

Bacteremia involving the "Streptococcus milleri" group: analysis of 19 cases.

During a 3-year study period, 19 patients at the University Hospital of Maastricht developed bloodstream infections with species of the "Streptococcus milleri" group, for an incidence of 0.33 per 1,000 admissions. The patients' median age was 48 years; the male-to-female ratio was 2.8. Eleven patients (57.9%) had underlying diseases, among which malignancy was predominant. Local trauma to the mucosal barrier was an important risk factor. An associated site of infection was found most frequently in the abdominal and thoracic cavities (nine and five cases, respectively). Bacteremia was polymicrobial in four of 19 episodes. The 20 infecting S. milleri strains were identified to the species level; Streptococcus anginosus was the most prevalent (16 strains). Eight strains carried Lancefield group C. The isolates were sensitive to most antibiotics. Abscess formation was documented in nine cases (47.3%); repeated drainage procedures were required in half of these episodes. Mortality was high (five of 19 patients, or 26.3%).

Adolescent↗

Interactions of fibronectin with streptococci: the role of fibronectin as a receptor for Streptococcus pyogenes.

Current evidence suggests that lipoteichoic acid (LTA) on the surface of Streptococcus pyogenes is centrally involved in the adherence of these bacteria to the oral mucosa. The major receptor on the surface of buccal mucosal cells for S. pyogenes is fibronectin. Fibronectin is a complex glycoprotein found in blood, extracellular matrices, and saliva. Many species of streptococci bind to fibronectin, although the characteristics of these interactions are different. Furthermore, whereas LTA is an efficient inhibitor of the binding of fibronectin to group A streptococci, it has little ability to inhibit the binding of fibronectin to Streptococcus pneumoniae or Staphylococcus aureus. Studies conducted with fibronectin show that different bacteria bind to different sites. The ability of various bacteria to interact with different domains on the fibronectin molecule may play an important role in bacterial adherence and tissue tropism.

Animals↗

Design of new benzoxazole-2-thione-derived inhibitors of Streptococcus pneumoniae hyaluronan lyase: structure of a complex with a 2-phenylindole.

The bacterial hyaluronan lyases (Hyals) that degrade hyaluronan, an important component of the extracellular matrix, are involved in microbial spread. Inhibitors of these enzymes are essential in investigation of the role of hyaluronan and Hyal in bacterial infections and constitute a new class of antibiotics against Hyal-producing bacteria. Recently, we identified 1,3-diacetylbenzimidazole-2-thione and related molecules as inhibitors of streptococcal Hyal. One of such compounds, 1-decyl-2-(4-sulfamoyloxyphenyl)-1-indol-6-yl sulfamate, was co-crystallized in a complex with Streptococcus pneumoniae Hyal and its structure elucidated. The resultant X-ray structure demonstrates that this inhibitor fits in the enzymatic active site via interactions resembling the binding mode of the natural hyaluronan substrate. X-ray structural analysis also indicates binding interactions with the catalytic residues and those of a catalytically essential hydrophobic patch. An IC50 value of 11 microM for Hyal from Streptococcus agalactiae (strain 4755) qualifies this phenylindole compound as one of the most potent Hyal inhibitors known to date. The structural data suggested a similar binding mode for N-(3-phenylpropionyl)-benzoxazole-2-thione. This new compound's inhibitory properties were confirmed resulting in discovery of yet another Hyal inhibitor (IC50 of 15 microM). These benzoxazole-2-thiones constitute a new class of inhibitors of bacterial Hyals and are well suited for further optimization of their selectivity, potency, and pharmacokinetic properties.

Benzoxazoles↗