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Species distribution of non-group D alpha-hemolytic streptococci in maternal genital and neonatal blood cultures.

At our hospital (Jefferson Davis Hospital, Houston, Tex.) since 1979, non-group D alpha-hemolytic streptococci have been isolated with increasing frequency from neonatal blood cultures with clinical findings of sepsis. A total of 47 such isolates were identified to the species level by the scheme of Facklam and were compared with 57 genital isolates from 167 maternity patients. Among the genital isolates, S. sanguis II and S. MG-intermedius accounted for 53 and 28%, respectively, and both were significantly less common in neonatal cultures (23 and 11%, respectively; P less than 0.05). Among neonatal isolates, S. mitis was the single most frequent species (35%), in contrast to its rare occurrence in maternal cultures (3.4%; P less than 0.001). The disparity between the prevalence of S. mitis in neonatal compared with maternal cultures suggests that this species of non-group D alpha-hemolytic streptococci may have increased virulence in neonatal hosts.

Female↗

Subgingival temperature (III). Relation to microbial counts.

The present investigation examined the relationship of selected bacterial species and subgingival temperature. 35 subjects were measured at 6 sites per tooth for clinical parameters and subgingival temperature. Measurements were repeated for 21 subjects at 2 month intervals providing a total of 66 subjects visits. At each visit, subgingival plaque samples were taken from the mesial aspect of each tooth and anaerobically dispersed, diluted and plated on non-selective media. After anaerobic incubation, colonies were lifted to nylon filters and specific species detected using digoxigenin-labeled whole chromosomal DNA probes. Species enumerated were; A. actinomycetemcomitans serotypes a and b, B. forsythus, B. gingivalis, B. intermedius I and II, C. ochracea, F. nucleatum ss. vincentii, P. micros, S. intermedius, S. sanguis I and II, V. parvula and W. recta. Total viable counts and counts of Capnocytophaga sp. were determined directly from the primary isolation plates. A total of 1581 samples were evaluated. Subject visits with higher mean subgingival temperatures had significantly higher mean %s of B. intermedius I and P. micros, and lower mean %s of Capnocytophaga sp. Sites with higher subgingival temperatures had elevated proportions of B. intermedius I and II, A. actinomycetemcomitans serotype a and B. gingivalis more frequently than sites with lower temperatures, while Capnocytophaga sp. were elevated more often at cooler sites. 43 of the subject visits had follow up attachment level measurements at 2 months. The 1026 microbial samples and the subgingival temperature measurements from these visits were related to longitudinal attachment change.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Effect of 6 months use of a dentifrice and oral rinse containing sanguinaria extract and zinc chloride upon the microflora of the dental plaque and oral soft tissues.

This study documented the effect upon the oral flora of twice daily brushing with a dentifrice containing 0.075% sanguinaria extract and 2% zinc chloride, followed by use of a mouthrinse containing 0.03% sanguinaria extract and 0.2% zinc chloride. Sixty subjects were randomly assigned to treatment or placebo groups and monitored in a 6-month double-blind clinical trial. Bacteriological samples from the tongue, buccal mucosa, and supra- and subgingival plaque were characterized at 0, 14, and 28 weeks. Microbiological monitoring showed no increases in populations of yeast, staphylococci, coliform organisms, or Pseudomonas. Total Gram-negative counts in supragingival plaque samples decreased 83% in the active group compared to a 232% increase for the control group. Populations of B. intermedius in supragingival plaque were significantly lower in the active group at 3 months and significantly lower counts of Fusobacterium sp. were observed at 3 and 6 months. Results indicate that use of the test products did not promote opportunistic overgrowth of pathogens in the oral flora. Additionally, the alterations in organisms associated with gingivitis may account for reductions in gingivitis seen in the active group.

Adult↗

Relation of counts of microbial species to clinical status at the sampled site.

The purpose of the present investigation was to relate clinical characteristics at a site to the frequency of detection, absolute counts and proportions of 14 subgingival species. Subgingival plaque samples were removed by curette from the mesial surface of 2299 teeth in 3 healthy and 87 subjects with periodontal attachment loss. Samples were dispersed, diluted and plated on Trypticase soy agar supplemented with 5% sheep blood. After 7 days of anaerobic incubation, colonies were lifted onto nylon filters, lysed and the DNA fixed to the filters. Digoxygenin-labeled DNA probes were used to identify colonies of each test species. Measurements of pocket depth, attachment level, recession, redness, bleeding on probing and suppuration were made at each sampled site. Total viable counts at sites ranged from 10(3) to greater than 10(8) and were strongly related to pocket depth. Mean total counts at sites less than 3 mm averaged 4.6 x 10(6), while mean counts at sites greater than 7 mm averaged 2.0 x 10(7). Species enumerated and % of sites colonized were as follows; V. parvula 44; S. sanguis II 36; B. intermedius I 33; C. ochracea 31; B. intermedius II 30; S. sanguis I 29; B. gingivalis 27; S. intermedius 25; P. micros 24; W. recta 23; F. nucleatum ss vincentii 18; B. forsythus 15; A. actinomycetemcomitans serotype a 10; A. actinomycetemcomitans serotype b 8. Counts of B. intermedius II were higher at sites which exhibited gingival redness while B. intermedius I was higher at sites which bled on probing. A. actinomycetemcomitans serotype b was more frequent and at higher mean % at sites without recession. The opposite was true for S. sanguis II. B. gingivalis was somewhat more prevalent and at higher levels at suppurating sites. B. gingivalis, B. intermedius I and II and B. forsythus were found more frequently and at higher levels at sites with deeper pockets, while V. parvula was less prevalent at sites with pocket depths less than 4 mm. B. gingivalis, B. intermedius I and A. actinomycetemcomitans serotype b increased with increasing pocket depth in both localized and widespread disease subjects, but mean counts were higher in the localized disease subjects at any pocket depth. Only W. recta was found at higher levels at deep sites in widespread disease subjects when compared with similar sites in localized disease subjects. No suspected pathogens were detected in 38% of shallow sites, 31% of intermediate sites and 22% of deep sites, 2/3 of deep pockets, but less than 1/2 of shallow pockets harbored at least 2 of the suspected pathogens.

Aggregatibacter actinomycetemcomitans↗

Streptococci and Actinomyces induce antibodies which cross react with epithelial antigens in periodontitis.

Perturbation of epithelial structure is a prominent but poorly understood feature of the immunopathological response to bacterial antigens which characterizes the destructive lesion of periodontitis. Western analysis of sera from 22 patients with periodontitis detected multiple antigens in extracts of epithelial cells whereas sera from 12 periodontally healthy subjects displayed only trace reaction with epithelial antigens. To investigate a possible relationship between the bacterial flora adjacent to diseased sites and the presence of antibodies reactive with epithelium, subgingival plaque samples were taken from deep periodontal pockets and cultured anaerobically. Gram positive bacteria containing antigens cross-reactive with epithelial cells were reproducibly isolated by probing membrane colony-lifts with affinity-isolated (epithelium-specific) antibodies and identified by 16S rDNA sequence homology as streptococci (S. mitis, S. constellatus and two S. intermedius strains) and Actinomyces (A. georgiae, and A. sp. oral clone). Conversely, when serum from patients with periodontitis was absorbed with the captured bacterial species the number of epithelial antigens recognized was specifically reduced. It was concluded that development of cross-reactive antibodies related to these organisms may contribute to perturbation of the epithelial attachment to the tooth and the progression of periodontitis. These autoreactive antibodies could also be a contributing factor in other diseases affecting epithelia.

Actinomyces↗

Identification of streptococci to species level by sequencing the gene encoding the manganese-dependent superoxide dismutase.

We have used a PCR assay based on the use of degenerate primers in order to characterize an internal fragment (sodA(int)) representing approximately 85% of the genes encoding the manganese-dependent superoxide dismutase in various streptococcal type strains (S. acidominimus, S. agalactiae, S. alactolyticus, S. anginosus, S. bovis, S. constellatus, S. canis, S. cricetus, S. downei, S. dysgalactiae, S. equi subsp. equi, S. equi subsp. zooepidemicus, S. equinus, S. gordonii, S. iniae, S. intermedius, S. mitis, S. mutans, S. oralis, S. parasanguis, S. pneumoniae, S. porcinus, S. pyogenes, S. salivarius, S. sanguis, S. sobrinus, S. suis, S. thermophilus, and S. vestibularis). Phylogenetic analysis of these sodA(int) fragments yields an evolutionary tree having a topology similar to that of the tree constructed with the 16S rRNA sequences. We have shown that clinical isolates could be identified by determining the positions of their sodA(int) fragments on the phylogenetic tree of the sodA(int) fragments of the type species. We propose this method for the characterization of strains that cannot be assigned to a species on the basis of their conventional phenotypic reactions.

Base Sequence↗

Effect of gels containing stannous fluoride on oral bacteria--an in vitro study.

The purposes of this investigation were to evaluate and compare the antimicrobial effect of (1) twelve 0.4 per cent stannous fluoride (SnF2) commercial products and (2) different concentrations of SnF2 (range = 0.02 to 3.28 per cent). The antibacterial inhibitory effect of various SnF2 gels was evaluated as to their effectiveness against oral plaque bacteria including strains of S. mutans, S. sanguis, S. sobrinus, A. viscosus, A. actinomycetemcomitans, and B. intermedius. When twelve different commercial preparations of 0.4 per cent SnF2 were compared for inhibitory effect on plaque bacteria, several of the SnF2 preparations were significantly more effective in inhibiting oral bacteria (p < 0.05). With increasing concentration of SnF2, there was a comparable increase in the inhibitory effect on the oral bacteria tested (r2 ranged from 0.867 to 0.996). SnF2 at a concentration of 0.4 per cent had a similar antibacterial effect to 0.12 per cent chlorhexidine. This in vitro study demonstrated that certain SnF2 products are highly effective in inhibiting the growth of bacteria often found in plaque, and this inhibitory effect is directly related to the concentration of the SnF2.

Actinomyces viscosus↗

A mixed-culture chemostat system to predict the effect of anti-microbial agents on the oral flora: preliminary studies using chlorhexidine.

A mixed-culture chemostat system, composed of nine bacterial species representative of plaque in health and disease, has been assessed as an improved laboratory method of evaluating the likely in vivo effects of antimicrobial agents used in dentistry. The advantages of the system include reproducibility, the long-term stable cultivation of bacteria under controllable conditions, and repeated sampling, for bacteriological and biochemical studies, without disrupting the stability of the community. The effects of (i) the continuous provision of chlorhexidine (CHX) and (ii) three pulses of CHX (final concentration in both experiments = 0.24 mmol/L) on the composition of the chemostat communities were monitored. Only L. casei survived the continuous provision of CHX; the other bacteria were killed and were lost at different rates which generally corresponded to their known sensitivities to CHX. After each CHX pulse, the numbers of bacteria fell markedly. Again, L. casei was least affected, while A. viscosus, B. intermedius, and F. nucleatum were temporarily undetectable but returned to their original levels within 2-4 generation times. Counts of S. mutans were affected more by CHX than those of S. sanguis or S. mitior. The effect of successive pulses of CHX on the viability of some bacteria and on acid production (as measured by pH-fall experiments) decreased, suggesting that adaptation to CHX had occurred. The fact that the in vitro observations paralleled previous clinical findings suggests that the mixed-culture system could be used as a predictive model of the probable effect on the oral flora of new anti-microbial agents prior to expensive trials in animals or human volunteers.

Actinomyces↗

In vitro adherence of microorganisms to denture base resin with different surface texture.

We examined the effects of various denture base resin surface textures on the adherence of microorganisms. S. sanguis and B. gingivalis adhered in greater amounts to the denture base resin than the other microorganisms tested. As to bacterial adherence according to polishing state, S. oralis, B. gingivalis C-101, and B. intermedius C-001 more adhered to the No. 400 paper-polished surface than to the buff-polished and smoothening-treated surfaces. S. sanguis less adhered to the smoothening-treated surface. S. mitis and C. albicans, on the other hand, more adhered to the smoothening-treated surface. For the other microorganisms tested, no relationship was observed between surface texture and bacterial adherence. The fall-off test revealed no remarkable differences in the fall-off of S. sanguis and B. gingivalis C-101 by the types of surface treatment. However, the fall-off of C. albicans was poorest from the No. 400 paper-polished surface. These results indicate that smoothening the denture base surface is important for denture plaque control.

Bacterial Adhesion↗

Microbiological findings in pregnant women with premature rupture of the membranes.

From 30 consecutive cases of premature rupture of the membranes (PROM) and matched controls, specimens from urine, cervix, amniotic fluid, and placenta as well as neonatal nose and throat swabs were investigated bacteriologically and virologically. In addition virus serological investigation was done. Among the PROM cases the anaerobic cervical isolates outnumbered the aerobic ones, and the total number of aerobic as well as anaerobic isolates was less in the control group. The anaerobes B. fragilis and Strept. intermedius were isolated from the cervix, amniotic fluid, or placenta in 23% and 30% of the PROM patients, respectively. None of the controls harboured B. fragilis, while Strept. intermedius was isolated from 6.7% of the controls. Group B streptococci were recovered from the mother's cervix in 20% of the PROM patients and in 6.7% among the controls. Four cases of neonatal septicaemia were encountered, and another two cases were clinically suspected, but not microbiologically verified, contributing to a high perinatal mortality rate (17,6%). Of the PROM patients, 27% developed puerperal infection, while none of the control mothers had such complications. The significance of the anaerobic bacteria as well as group B streptococci for the maternal and neonatal outcome in cases of PROM is discussed, and a possible aetiological role of ascending infection in this complication of pregnancy is postulated.

Adult↗

The use of DNA probes to examine the distribution of subgingival species in subjects with different levels of periodontal destruction.

The present investigation examined the distribution of 14 subgingival species at a total of 2299 sites in 90 subjects with different levels of periodontal destruction. Subgingival plaque samples taken from the mesial aspect of each tooth were anaerobically dispersed, diluted and plated on non selective media. After anaerobic incubation, colonies were lifted to nylon filters and specific species detected using digoxygenin-labeled whole chromosomal DNA probes. The mean total viable count for all sites in all subjects was 8.3 x 10(6). The probes accounted for an average of 27.8% of the total viable count. The % of subjects in which each species was detected was as follows; V. parvula, 98; B. intermedius I, 98; S. sanguis II, 96; B. intermedius II, 95; C. ochracea, 94; B. gingivalis, 91; S. sanguis I, 85; W. recta, 83; F. nucleatum ss. vincentii, 82; S. intermedius, 80; B. forsythus, 76; P. micros, 74; A. actinomycetemcomitans serotype a, 62 and A. actinomycetemcomitans serotype b, 52. The % of sites colonized by each of the 14 test species varied considerably within different subjects. The median number of sites colonized by different species ranged from 3.6% for A. actinomycetemcomitans serotype b to 43.5% for V. parvula. In half the subjects, the mean % of the total viable counts for each of the test species was less than 4%. When subjects were divided on the basis of % of sites at baseline with greater than 3 mm attachment loss, the 14 probes accounted for 29.9% of the microbiota in the localized disease group and 25% in the widespread disease group.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Subgingival microbial profile of Papillon-Lefèvre patients assessed by DNA-probes.

The prevalence of 18 selected bacterial species was assessed by means of "checkerboard" DNA-DNA hybridisation in a group of 12 Saudi-Arabian adolescents with Papillon-Lefèvre syndrome. A total of 36 tooth sites were investigated. The patients exhibited severe periodontal disease with deep pockets. All 12 patients harboured the putative bacterial pathogens P. intermedia, F. nucleatum, P. micros and S. intermedius while T. denticola, B. forsythus, P. nigrescens, E. corrodens, S. noxia and C. rectus were recovered from 11 patients. P. gingivalis was recovered from 9 patients and 18 sites while corresponding figures for A. actinomycetemcomitans were 8 and 19, respectively. A number of the investigated species (B. forsythus, T. denticola, P. intermedia, C rectus) reached high levels (> or =10(6) cells) in more than 1/2 of the patients. On the other hand, bacteria such as A. actinomycetemcomitans and P. gingivalis were infrequently encountered at high levels in these subgingival samples. In conclusion, the analysis failed to demonstrate a PLS-specific profile of the subgingival infection, since the bacterial composition of the sampled sites closely resembled that characterising deep pockets in adult periodontitis patients.

Adolescent↗

The effects of periodontal therapy on serum antibody (IgG) levels to plaque microorganisms.

The influence of periodontal therapy on serum antibody titers to selected periodontal disease-associated microorganisms was assessed in 23 patients having chronic inflammatory periodontal disease (CIPD). The immunoglobulin G (IgG) titers were determined by the microELISA technique in serum samples obtained prior to treatment; following a hygienic phase which included scaling, root planing, and oral hygiene instruction; following surgical treatment; and one year and two years following hygienic phase (maintenance phase). Considerable individual variability existed in the magnitude of immune response to specific bacterial preparations. Significant reductions in the mean antibody titers were seen to A. viscosus, S. sanguis, F. nucleatum, S. sputigena, B. gingivalis, B. intermedius, B. melaninogenicus, T. vincentii, and T. denticola by the end of the second year of maintenance. There was no consistent response to Capnocytophaga. When individual patient responses were examined, 6 of the 23 were found to have elevated titers to at least one of the microorganisms in the interval between pretreatment and the end of the hygienic phase; however, in all but one case, the titers at the end of the second year of maintenance were below pretreatment levels. Antibody levels to bacteria such as S. sanguis were modified during therapy. This would indicate that immune responses to microbes not generally considered to be "periodontal pathogens" may be modified by adjuvant activity associated with subgingival plaque or changes in the environment of the sulcus and that subsequent changes in titer do not necessarily reflect a role of that microorganism in the disease process.

Adult↗

Smoking and subgingival microflora in periodontal disease.

AIM: The present investigation was undertaken to analyze the influence of smoking on the periodontal disease associated subgingival microflora. The population included 33 smokers and 31 non-smokers in the age range 36-86 years. METHODS: Microbial samples were obtained from 4 sites per patient. The checker-board DNA-DNA hybridization technology was used for detection of the bacterial species P. gingivalis, P. intermedia, P. nigrescens, B. forsythus, A. actinomycetemcomitans, F. nucleatum, T. denticola, P. micros, C. rectus, E. corrodens, S. noxia and S. intermedius. RESULTS: Using score 1 as cutoff, contrasting colonized versus non-colonized patients, 8 out of 12 species were detected in > or = 90% of both smokers and non-smokers. Using score 4 as cutoff, contrasting heavily colonized patients versus non-colonized and less heavily colonized patients, the detection rates decreased in both smokers and non-smokers. No significant differences in detection rates were observed between smokers and non-smokers. Logistic regression analysis indicated that neither smoking, probing depth nor gingival bleeding influenced the occurrence of the species analyzed. The lack of a smoking exposure dose-response further supported the indication of a limited influence of smoking. CONCLUSION: Smoking exerts little, if any, influence on the subgingival occurrence of several of the bacteria most commonly associated with periodontal disease.

Adult↗

Group milleri streptococci: significant pathogens in solid organ recipients.

Group milleri streptococci (GMS) comprise a heterogeneous group of streptococci including the species intermedius, constellatus and anginosus. They may cause chronic intra-abdominal and intrathoracic abscesses, which are difficult to treat. This is a retrospective analysis including 45 transplant recipients in whom GMS were isolated. The epidemiology, clinical significance and the impact on the outcome in all transplant patients with infections caused by GMS during a 4-year period (2001-2004) was evaluated. The 45 solid organ recipients (88 isolates) included 34 liver-, four kidney/pancreas-, one kidney-, two small bowel-, three combined liver/kidney- and one combined kidney/small bowel transplant recipient. In 42 cases GMS caused intra-abdominal infection, in two cases pleural empyema and in one case soft tissue infection. Only a single isolate of GMS was cultured from blood. In 54 of the 88 specimens (61%), which grew GMS, other pathogens were also isolated. GMS frequently caused recurrent cholangitis (n = 17) associated with anastomotic and nonanastomotic biliary strictures. These cases were managed by repeated stenting or surgical intervention and prolonged antibiotic therapy. No patient died directly related to GMS infection and all except one case responded to combined surgical/antibiotic treatment. One pancreas graft was lost because of erosion haemorrhage associated with an abscess. GMS were susceptible to penicillin G, carbapenems and clindamycin, whereas cephalosporins and quinolones showed intermediate activity or resistance in some cases, and GMS in general were found resistant to aminoglycosides. GMS may cause serious infections in transplant recipients which are difficult to treat. Their prevalence in transplant surgical site infections thus far may have been underestimated.

Anti-Bacterial Agents↗

Utilization of sialic acid by viridans streptococci.

The importance of viridans streptococci as agents of serious extra-oral diseases, including endocarditis, is now recognized. We have tested the hypothesis that the ability to utilize sialic acid as a nutrient source may play a role in the proliferation of these organisms. The type strains of the 15 presently recognized species of viridans streptococci and two clinical isolates-S. oralis (AR3), isolated from a patient with infective endocarditis, and S. intermedius (UNS35), a brain abscess isolate-were studied for their ability to utilize sialic acid. Only S. oralis, S. sanguis, S. gordonii, S. mitis ("oralis group") S. intermedius, S. anginosus, S. constellatus ("milleri group"), and S. defectivus ("nutritionally variant group") were able to use sialic acid (N-acetylneuraminic acid) efficiently as a sole carbon source. Formate, acetate, and ethanol were produced as the major metabolic end-products of sialic acid metabolism, while corresponding glucose-grown cultures produced lactate as the major metabolic end-product. Utilization of sialic acid was independent of the production of sialidase. Cell-free extracts of sialic acid-grown cultures expressed elevated levels of N-acetylneuraminate pyruvate-lyase (NPL; the first enzyme in the intracellular catabolism of sialic acid) and N-acetylglucosamine-6-phosphate (GlcNAc-6-P) deacetylase and glucosamine-6-phosphate (GlcN-6-P) deaminase (enzymes involved in the intracellular catabolism of N-acetylglucosamine). These activities were repressed by growth in the presence of glucose. The intracellular fate of sialic acid, after cleavage by NPL into N-acetylmannosamine (ManNAc) and pyruvate, is uncertain, but the elevated levels of GlcNAc-6-P deacetylase and GlcN-6-P deaminase in sialic acid-grown cells suggest that phosphorylation and isomerization are possible steps in the metabolism of ManNAc to generate an intermediate common to the pathway of N-acetylglucosamine metabolism. The species of viridans streptococci that have the ability to utilize sialic acid are those most commonly associated with extra-oral diseases, and this ability is likely to play a role in the persistence and survival of these infecting organisms in vivo.

Acetylglucosamine↗