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Vancomycin tolerance, a potential mechanism for refractory gram-positive bacteremia observational study in patients with cancer.

BACKGROUND: The clinical significance of infections caused by vancomycin-tolerant (Vt) gram-positive organisms in patients with cancer remains unclear. METHODS: Twenty-five patients with nonenterococcal gram-positive bloodstream infection, which was refractory to vancomycin therapy, were identified by reviewing the Infectious Diseases consultation database at the tertiary care cancer center. Among these, 8 patients in whom vancomycin-tolerance was documented are described. Antibiotic tolerance was defined as a > 32 times increase in minimum bactericidal concentration compared with minimum inhibitory concentration. RESULTS: Eight patients with persistent fever and bacteremia of > 72 hours' duration after the initiation of vancomycin therapy were treated. The median age of these patients, which included 3 men and 5 women, was 44 years +/- 11 years. Solid tumors were more common (6 patients) and 2 patients had acute leukemia. Six patients (75%) were neutropenic (absolute neutrophil count < 500/mm3), including 2 breast cancer patients who had undergone autologous stem cell transplantation. The causative organisms were Staphylococcus aureus (n = 3 patients), group G streptococci (n = 2 patients), and Staphylococcus epidermidis, Streptococcus mitis, and Streptococcus sanguis (1 patient each). All isolates demonstrated a minimum bactericidal concentration for vancomycin that was at least 32 times greater than the minimum inhibitory concentration. Rapid defervescence (< or = 24 h) and resolution of bacteremia occurred with the addition of gentamicin (4 patients) or gentamicin plus rifampin (4 patients). None of these infections recurred after discontinuation of therapy. CONCLUSIONS: Lack of clinical and/or microbiologic response to vancomycin should raise the suspicion of possible infection due to Vt gram-positive bacteria, and alternative bactericidal therapy should be instituted early, especially in patients with underlying immune suppression.

Adult↗

Prophylactic co-trimoxazole versus norfloxacin in neutropenic children--perspective randomized study.

Co-trimoxazole or norfloxacin were randomly administered to 44 granulocytopenic children with malignancies in order to prevent bacterial infections. Although more patients in the co-trimoxazole group had febrile episodes (p less than 0.01), the mean of febrile days and the mean of days with systemic antibiotics did not differ significantly in the two groups. Five patients in the co-trimoxazole group had a microbiologically documented infection (four with septicemia) due to Escherichia coli (n = 2), Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus sp. There were four septicemic episodes in the norfloxacin group due to P. aeruginosa, Streptococcus pneumoniae, Streptococcus mitis and Streptococcus faecalis. Compliance was good during administration of both drugs. No signs or symptoms of arthropathy were seen in the norfloxacin group. The number of gram-negative bacilli resistant to co-trimoxazole isolated from stools significantly increased during prophylaxis with co-trimoxazole (p less than 0.001). Norfloxacin did not select resistant strains and was very active in eradicating gram-negative bacilli from stools (27.5% of positive cultures).

Adolescent↗

Exogenous thymidine and reversal of the inhibitory effect of sulfamethoxazole-trimethoprim on streptococci.

The practice of using sulfamethoxazole-trimethoprim (SXT) for the selective isolation of Streptococcus pyogenes and as a taxonomic character in the presumptive identification of streptococci was applied to 17 strains of different groups of streptococci to determine their characteristic behaviour in the presence of exogenous thymidine. Streptococcus pyogenes, Streptococcus agalactiae and group D enterococci utilized thymidine, the first two species obtaining a maximum reversal of the inhibitory effect of SXT at thymidine concentrations of 1.2 micrograms/ml and 0.6 micrograms/ml or higher, respectively. For group D enterococci, the degree of reversal of the inhibitory effect was proportional to the thymidine concentration. In contrast, the four viridans species studied (Streptococcus sanguis I, Streptococcus salivarius, Streptococcus mitis and Streptococcus sanguis II) and Streptococcus pneumoniae were unable to utilize thymidine from an exogenous source and thus growth remained inhibited even at the highest concentrations of thymidine tested. For selective isolation and identification of streptococci only stable media with batch-to-batch consistency are recommended together with a known quantity of thymidine.

Agar↗

Bacteremia due to viridans streptococci in neutropenic patients: a review.

Viridans streptococci have long been considered, with the exception of the ability to cause endocarditis, as minor pathogenic agents. More recently, however, these bacteria have become a major concern in neutropenic patients undergoing a chemotherapeutic treatment. In this high-risk population, they can be responsible for up to 39% of bacteremia cases and are the most frequent cause of this type of infection. The most frequently isolated species in blood cultures are Streptococcus mitis and Streptococcus sanguis II. Viridans streptococcus bacteremia can be accompanied by serious complications, like adult respiratory distress syndrome (ARDS) (3% to 33%), shock (7% to 18%) or endocarditis (7% to 8%). Mortality rates range from 6% to 30%. Case-control studies have identified the following risk factors: severe neutropenia (< 100 neutrophils/mm3), prophylactic antibiotic treatments with quinolone or co-trimoxazole, absence of intravenous antibiotics at the time of bacteremia, high doses of cytosine arabinoside, oropharyngeal mucositis, and heavy colonization by viridans streptococci. The introduction of penicillin in prophylactic antibiotic treatments has reduced the incidence of these infections, but the long-term use of penicillin could be compromised by the emergence of resistant strains.

Bacteremia↗

Viridans streptococci isolated from the bloodstream. Relevance of species identification.

Over a 4-year-period, 104 isolates belonging to the viridans streptococci were recovered from the blood and identified to the species according to Beighton and co-workers. Streptococcus oralis was the species most frequently recovered from patients in the hematology unit [29 of 39 (74%)]. Streptococcus mitis ranked second [seven of 39 (18%)]. Both species were associated with oromucositis. Isolates presently identified as S. oralis are mainly those previously identified as S. mitis or Streptococcus sanguis II. Streptococcus milleri was most frequently isolated in the patients from the general hospital population [28 of 65 (43%) isolates]: Streptococcus anginosus (n = 20), Streptococcus constellatus (n = 4), and Streptococcus intermedius (n = 4). In 14 episodes from the general hospital population, clinical significance was judged questionable. From this part of the laboratory, clinical significance could not be predicted from the number of blood cultures grown, nor from the delay of growth detection. The Rapid ID 32 Strep system agreed well with the identifications according to the scheme of Beighton et al., whereas the API 20 Strep system did not.

Adolescent↗

Binding of oral streptococci to human fibrinogen.

The interaction of oral streptococci with human fibrinogen was investigated. Streptococcus gordonii was chosen as a representative species to study the binding to fibrinogen. S. gordonii DL1 adhered to immobilized fibrinogen and bovine serum albumin. Binding to immobilized fibrinogen was saturable, concentration and temperature dependent. The binding of S. gordonii DL1 to fibrinogen was inhibited by anti-fibrinogen antibody. Heating of the bacteria for 1 h at 95 degrees C resulted in 90% inhibition of the binding. Trypsin treatment of the bacteria resulted in decreased binding. Neither lipoteichoic acid nor culturing of the bacteria in a sucrose-supplemented medium had any effect on the binding. S. gordonii, Streptococcus sanguinis, Streptococcus mitis, and Streptococcus oralis bound to the immobilized fibrinogen, but mutans streptococci did not. None of the oral streptococci tested bound to the fibrinogen in fluid phase. These results suggest that the binding of S. gordonii DL1 to immobilized fibrinogen is mediated through a specific protein adhesin-receptor interaction, and fibrinogen acts as a cryptitope.

Adhesins, Bacterial↗

A global gene pool for high-level cephalosporin resistance in commensal Streptococcus species and Streptococcus pneumoniae.

Highly penicillin- and cephalosporin-resistant Streptococcus mitis and Streptococcus oralis were isolated in Spain, Hungary, and Berlin. With chromosomal DNA of these strains, resistant transformants of Streptococcus pneumoniae were obtained that expressed low-affinity variants of penicillin-binding proteins (PBPs) 2x, 1a, 2a, and 2b in different combinations, depending on the selective conditions. The transformants had cefotaxime MICs of up to 6 microg/mL, and those with a low-affinity PBP 2b were highly deficient in penicillin-induced lysis. Sequence analysis of the pbp2x genes confirmed the presence of a global gene pool of penicillin resistance determinants shared by commensal and pathogenic streptococci.

Amino Acid Sequence↗

Autolysin-targeted LightCycler assay including internal process control for detection of Streptococcus pneumoniae DNA in clinical samples.

The development and clinical evaluation of a LightCycler PCR assay, including an internal process control (IPC), to detect the Streptococcus pneumoniae autolysin gene in clinical samples is reported. The assay was developed to provide a second target for use in conjunction with existing pneumolysin PCR assays to increase the reliability of non-culture PCR diagnosis of pneumococcal infection. Primers amplify a 173 bp fragment of the autolysin gene (lytA), which is detected by fluorescence-labelled hybridization probes. An IPC was designed to check for the presence of PCR inhibitors and loss of assay sensitivity. The IPC product was amplified by the lytA primers and detected by a second set of hybridization probes. The analytical specificity of the autolysin PCR assay was 100% against 39 other bacterial species tested; these included related streptococci and other organisms. The assay, which could reliably detect 50 fg purified pneumococcal DNA per reaction, was capable of distinguishing between S. pneumoniae and atypical Streptococcus mitis and Streptococcus oralis strains known to contain the lytA gene. Using DNA extracts from a panel of EDTA bloods from patients with blood-culture-confirmed pneumococcal infection, the autolysin PCR had a sensitivity of 42.9%, which was similar to a previously reported TaqMan pneumolysin PCR (43.8%) run in parallel. Total agreement was shown between the autolysin assay and the pneumolysin TaqMan assay when used to test 23 culture-negative clinical samples, of which eight were positive by PCR, adding valuable clinical information. A specific autolysin-based LightCycler assay has been developed to complement pneumolysin PCR for the detection of S. pneumoniae in clinical samples. This should be a particularly useful tool for the rapid and sensitive diagnosis of pneumococcal meningitis, even after an antibiotic has been administered. However, poor sensitivity on blood samples limits its usefulness in other bacteraemic infections.

DNA, Bacterial↗

Serum antibody responses to Streptococcus mutans antigens in humans systemically infected with oral streptococci.

Sera from patients with subacute bacterial endocarditis (SBE) due to Streptococcus mutans or other oral streptococci and from normal subjects were assayed by enzyme-linked immunosorbent assay for antibodies to defined S. mutans antigens. Antibodies of IgG and IgA isotypes to Ag I/II and Ag III were greatly elevated in S. mutans-SBE sera, and the IgA antibodies in 3 sera included both polymeric and monomeric forms. Elevated IgM and IgG anti-lipoteichoic acid and IgG and IgA anti-serotype c polysaccharide antibodies were also found. The sera of 4 of 6 patients infected with other oral streptococci also displayed antibodies to S. mutans Ag I/II. Sera of 3 patients infected with Streptococcus mitis or Streptococcus oralis, but none of the S. mutans-infected cases, showed elevated antibodies to human heart sarcolemma, and all SBE sera had elevated rheumatoid factor. These results suggest that the known surface protein antigens of S. mutans are immunodominant in humans, and are not likely to be heart cross-reactive.

Antibodies, Bacterial↗

Purification and properties of a bacteriocin of Staphylococcus epidermidis isolated from dental plaque.

An extracellular bacteriocin of Staphylococcus epidermidis isolated from dental plaque was purified and characterized. Its molecular mass was 3500 Da and pI was 10.5. This bacteriocin inhibited the growth of Staphylococcus aureus, Streptococcus salivarius, and Streptococcus mitis, but Streptococcus mutans, Streptococcus sanguis and other oral indigenous bacterial species examined were not inhibited. The mode of inhibition was found to be bacteriostatic.

Bacterial Proteins↗

Comparison of the subgingival microbiota of periodontally healthy and diseased adults in northern Cameroon.

Our study is the 1st report on subgingival microbiota in adult Cameronians. The aim was to investigate, using the checkerboard DNA-DNA hybridization technique, the prevalence of 18 oral species in subgingival plaque samples obtained from sex- and age-matched Cameronian adults with and without periodontal destruction. We also compared cultivation and the Affirm DP test with the checkerboard technique in their capability to detect some selected species among the 18. 21 adult periodontitis patients and 21 periodontally healthy subjects were examined and the results were compared statistically. Each periodontitis patient had at least 4 pockets of > or = 6 mm depth, while the healthy subjects had no sites more than 3 mm deep. Results of the checkerboard analysis showed that significantly (p < 0.05) more periodontitis patients tested positive for most of the 18 bacterial species. The Gram-positive species Actinomyces naeslundii, Streptococcus mitis and Streptococcus sanguis, known as microbiota of healthy sites, were detected significantly more frequently in the healthy group. Cultivation demonstrated P. gingivalis, B. forsythus, A. actinomycetemcomitans, P. intermedia and F. nucleatum in significantly lower %s of patients as compared to the checkerboard technique. Furthermore, the Affirm DP test detected P. gingivalis and B. forsythus in significantly fewer patients than did the checkerboard technique. A. actinomycetemcomitans was detected in 52.3% of the patients by the latter technique while the Affirm DP test failed to detect the bacterium in any of the samples. Overall, the results of the present study confirm the importance of the screening method and indicate that the prevalences of the investigated putative periodontal pathogens and beneficial species in the healthy and diseased adult Cameronians were similar to those reported for adults in the West and in some developing countries.

Actinomyces↗

Microbiota of gingivitis in man.

A study on the predominant cultivable microorganisms inhabiting gingival crevices affected with a chronic gingivitis was carried out using the roll tube culture technique. Samples were obtained from nine individuals 25--42 years of age. Gram-positive rods make up 29.1% of the isolates and included mainly Actinomyces naeslundii, Actinomyces israelii, and Actinomyces viscosus. Streptococcus mitis and Streptococcus sanguis together made up 26.8% of the cultivable organisms. Peptostreptococcus averaged 3.0% of the organisms recovered. Gram-negative anaerobic rods constituted 25.0% of the total isolates with Fusobacterium nucleatum, Bacteroides melaninogenicus ss. intermedius. Bacteriodes ochraceus, other Bacteroides species, Selenomonas sputigena, and Campylobacter sputorum as the most predominant isolates. Haemophilus parainfluenzae averaged about 14% and Veillonella species 4.3% of the cultivable microflora. The data presented indicate that the subgingival microflora of a chronic gingivitis differs from those of healthy periodontium and advanced adult and juvenile periodontis. This might suggest that different infectious processes may be operative in various clinical entities of periodontal disease.

Actinomyces↗

Degradative enzymes of oral streptococci.

Members of the Streptococcus sanguis group (SSG) and Streptococcus milleri group (SMG) were screened for their ability to produce glycosidase, arylamidase (peptidase), protease, dextranase and glycosyltransferase activities. Species within each group produced unique patterns of activity. The most commonly produced glycosidases were beta-D-glucosidase, beta-D-galactosidase, N-acetyl-beta-D-glucosaminidase and N-acetyl-beta-D-galactosaminidase and the least commonly produced glycosidase activity was beta-fucosidase with Streptococcus intermedius (SMG) being the only species capable of producing the activity. For arylamidase activity, the most commonly produced type was lysine-arylamidase. Glycosidase and arylamidase activities were localized to particular sub-cellular fractions. alpha-galactosidase was found only in culture supernatant fluids whereas N-acetyl-beta-D-glucosaminidase was found in all fractions; the culture supernatant, cell wall, cell membrane and cytoplasm. No arylamidase activity was seen in culture supernatants. Phe-arg-arylamidase was found only in cytoplasmic fractions whereas val-pro-argarylamidase was found in cell walls, cell membranes and cytoplasmic fraction. Protease activity was measured as the degradation of bovine serum albumin (BSA) and casein. Casein was degraded by a number of strains whereas no species/strains were able to degrade BSA. Streptococcus intermedius, Streptococcus constellatus (SMG), Streptococcus mitior and Streptococcus defectivus (SSG) were the only species that produced hyaluronidase and no species produced chondroitin sulphatase. The groups were also examined for their abilities to produce glycosyltransferase and dextranase. Strep. sanguis, Streptococcus gordonii, Streptococcus mitis and Streptococcus oralis produced glucosyltransferase and, with the exception of the latter species, fructosyltransferase. No species within the SMG was capable of producing either glycosyltransferase.(ABSTRACT TRUNCATED AT 250 WORDS)

Aminopeptidases↗

Viridans group streptococci are donors in horizontal transfer of topoisomerase IV genes to Streptococcus pneumoniae.

A total of 46 ciprofloxacin-resistant (Cip(r)) Streptococcus pneumoniae strains were isolated from 1991 to 2001 at the Hospital of Bellvitge. Five of these strains showed unexpectedly high rates of nucleotide variations in the quinolone resistance-determining regions (QRDRs) of their parC, parE, and gyrA genes. The nucleotide sequence of the full-length parC, parE, and gyrA genes of one of these isolates revealed a mosaic structure compatible with an interspecific recombination origin. Southern blot analysis and nucleotide sequence determinations showed the presence of an ant-like gene in the intergenic parE-parC regions of the S. pneumoniae Cip(r) isolates with high rates of variations in their parE and parC QRDRs. The ant-like gene was absent from typical S. pneumoniae strains, whereas it was present in the intergenic parE-parC regions of the viridans group streptococci (Streptococcus mitis and Streptococcus oralis). These results suggest that the viridans group streptococci are acting as donors in the horizontal transfer of fluoroquinolone resistance genes to S. pneumoniae.

Base Sequence↗

Bacterial aggregating activity in human saliva: comparisons of bacterial species and strains.

Using a quantitative assay to measure saliva-mediated bacterial aggregating activity, we have surveyed 20 streptococcal strains with saliva samples obtained from a large population study. Individual saliva samples demonstrated characteristic levels of aggregating activity for Streptococcus sanguis M5. In general, high activity for this strain was associated with high activity for other strains of S. sanguis. Streptococcus mitis, and Streptococcus salivarius. The population distribution of aggregating activity for Streptococcus mutans, however, was different.

Humans↗

Characteristic differences between saliva-dependent aggregation and adhesion of streptococci.

Comparison of saliva-mediated aggregation of Streptococcus sanguis, Streptococcus mitis, and Streptococcus mutans and adhesion of these organisms to saliva-coated hydroxyapatite showed that there was no relationship between these two activities. Adsorption of salivary aggregating activity to bacteria appears to have little effect on the ability of the residual saliva to support adherence; conversely, adsorption of salivary adherence factors to hydroxyapatite does not affect aggregation. Although heating saliva significantly reduces bacterial aggregation, it has little or no effect on adherence. A comparison of aggregation and adhesion with serial dilutions of saliva demonstrated that adhesion could still be detected at 100 to 500-fold-lower concentrations of salivary protein that bacterial aggregation. These findings support the concept that aggregation and adherence involve two distinct mechanisms of microbial clearance in the oral cavity.

Adhesiveness↗

Antigenic relationships among immunoglobulin A1 proteases from Haemophilus, Neisseria, and Streptococcus species.

To investigate the antigenic variation and relationships of immunoglobulin A1 (IgA1) proteases among different species and genera, we examined a comprehensive collection of serine type and metallo-type IgA1 proteases and corresponding antisera in enzyme neutralization assays. Sharing of neutralizing epitopes of metallo-type IgA1 proteases from Streptococcus pneumoniae, Streptococcus sanguis, Streptococcus mitis, and Streptococcus oralis and of serine type IgA1 proteases from Haemophilus and pathogenic Neisseria species was extremely limited. A number of limited to strong cross-reactions in such epitopes were found among serine type IgA1 proteases released by members of the genera Haemophilus and Neisseria, reflecting the common origin of their iga gene. However, the relatively limited prevalence of shared "neutralizing" epitopes of IgA1 proteases from the two genera indicates that they rarely induce immunity to each other. In contrast, extensive sharing of neutralizing epitopes was found between N. meningitidis and N. gonorrhoeae IgA1 proteases, making them potentially attractive vaccine components. Among metallo-type IgA1 proteases, several pneumococcal proteases were found to induce neutralizing antibodies to IgA1 proteases of oral streptococci whereas the opposite was not the case.

Animals↗

Oral streptococci with genetic determinants similar to the glucosyltransferase regulatory gene, rgg.

The Streptococcus gordonii Challis glucosyltransferase structural gene, gtfG, is positively regulated by the upstream gene, rgg, the only described gtf regulatory determinant in oral streptococci. Southern hybridization analyses indicated that rgg-like and gtfG-like determinants were present on the same HindIII fragment in strains of S. gordonii, Streptococcus sanguis, and Streptococcus oralis, whereas no rgg-like determinants were detected in mutans streptococci, Streptococcus mitis, and Streptococcus salivarius.

Base Sequence↗