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Coaggregation of oral Bacteroides species with other bacteria: central role in coaggregation bridges and competitions.

Seventy-three freshly isolated oral strains representing 10 Bacteroides spp. were tested for their ability to coaggregate with other oral gram-negative and gram-positive bacteria. None coaggregated with any of the gram-negative strains tested, which included Capnocytophaga gingivalis, C. ochracea, C. sputigena, and Actinobacillus actinomycetemcomitans. Strains of Bacteroides buccae, B. melaninogenicus, B. oralis, and B. gingivalis failed to coaggregate with any of the gram-positive strains tested. However, six Bacteroides spp. coaggregated with one or more species of gram-positive bacteria. Most isolates of B. buccalis, B. denticola, B. intermedius, B. loescheii, B. oris, and B. veroralis coaggregated with strains of Actinomyces israelii, A. viscosus, A. naeslundii, A. odontolyticus, Rothia dentocariosa, or Streptococcus sanguis. The strongest coaggregations involved B. denticola, B. loescheii, or B. oris; 22 of 25 strains coaggregated with A. israelii. Only B. loescheii interacted with certain strains of S. sanguis; these coaggregations were lactose inhibitable and were like coaggregations between A. viscosus and the same strains of S. sanguis. In fact, B. loescheii and A. viscosus were competitors for binding to S. sanguis. Many bacteroides also acted as coaggregation bridges by mediating coaggregations between two noncoaggregating cell types (e.g., S. sanguis and A. israelii). Evidence for binding-site competition and coaggregation bridging involving noncoaggregating cell types from three different genera provides support for the hypothesis that these intergeneric cell-to-cell interactions have an active role in bacterial colonization of the oral cavity.

Actinomyces↗

Value of the L-pyrrolidonyl-beta-naphthylamide hydrolysis test for identification of select gram-positive cocci.

A 4 hr test for hydrolysis of L-pyrrolidonyl-beta-naphthylamide (PYR) was evaluated for its use in identifying group A beta-hemolytic streptococci, group D enterococci, and Staphylococcus haemolyticus. All of the group A streptococci and the enterococci tested were positive in the PYR test, whereas all of the nongroup A, B, or D beta-hemolytic streptococci, all of the viridans, and all of the Streptococcus pneumoniae isolates tested were PYR negative. Two strains of Aerococcus species were PYR positive and could not be differentiated from enterococci in this test. All strains of S. haemolyticus, a single strain of S. intermedius and two of seven strains of S. warneri were PYR positive whereas all other staphylococci tested were PYR negative. The PYR test is suitable for routine, presumptive identification of group A streptococci, group D enterococci and S. haemolyticus in the clinical laboratory.

Humans↗

Classification and identification of the viridans streptococci.

This review traces the history of the human, nonhemolytic, or viridans, streptococci and describes improvements in their taxonomy wrought by study of their biochemical profiles and analysis of their nucleic acids. The goal was to define species on the basis of genetic relationships and to describe these species by their phenotypic characteristics so that they can be easily identified. This method has resulted in the division of some species. Streptococcus mutans has been divided into four species, two of which are common in humans. Three more mutans group species are indigenous to animals. Conversely, S. constellatus, S. intermedius, and "S. milleri" have been combined under S. anginosus. S. mitis (or "S. mitior") can be well-defined and includes S. sanguis II. There is genetic heterogeneity within S. sanguis, but the species is usually easy to identify. There is also some heterogeneity in S. bovis, but most human isolates are genetically related. Discussions of the taxonomy of these species are accompanied by descriptions of the characteristics by which these streptococci can be identified. Among these species are potential pathogens which should be suspected in cases of endocarditis and purulent infections of liver, brain, and other tissues.

Animals↗

Bacteriologic characteristics and antimicrobial susceptibility of 70 clinically significant isolates of Streptococcus milleri group.

The phenotypical characteristics of 70 clinical isolates of Streptococcus milleri group were analyzed. Association of the three species with particular sites of isolation could not be demonstrated in all cases, but S. anginosus strains predominated in abdominal area, while S. intermedius was isolated mainly in hepatic abscesses. Penicillin G and other beta-lactams showed good in vitro activity against these streptococci, whereas > 10% exhibited resistance to erythromycin and clindamycin. Resistance to tetracycline reached almost 40%. All of the isolates were uniformly susceptible to vancomycin, chloramphenicol, and trimethoprim.

Drug Resistance, Microbial↗

"Streptococcus milleri" endocarditis caused by Streptococcus anginosus.

Unlike other viridans streptococci, members of the "Streptococcus milleri group" are often associated with abscess formation, but are only rare causes of infective endocarditis. Although it has been shown that almost all S. intermedius isolates and most S. constellatus isolates, but only 19% of S. anginosus isolates, were associated with abscess formation, no report has addressed the relative importance of the 3 species of the "S. milleri group" in infective endocarditis. During a 5-year period (April 1997 through March 2002), 6 cases of "S. milleri" endocarditis (out of 377 cases of infective endocarditis), that fulfil the Duke's criteria for the diagnosis of infective endocarditis, were encountered. All 6 "S. milleri" isolates were identified as S. anginosus by 16S ribosomal RNA (rRNA) gene sequencing. Three patients had underlying chronic rheumatic heart disease and 1 was an IV drug abuser. Five had monomicrobial bacteremia, and 1 had polymicrobial (S. anginosus, S. mitis, Granulicatella adiacens, and Slackia exigua) bacteremia. Two patients died. None of the 6 isolates were identified by the Vitek system (GPI) or the API system (20 STREP) at >95% confidence. All 6 isolates were sensitive to penicillin G (MIC 0.008-0.064 microg/mL), cefalothin, erythromycin, clindamycin, and vancomycin. Accurate identification to the species level, by 16S rRNA gene sequencing, in cases of bacteremia caused by members of the "S. milleri group", would have direct implication on the underlying disease process, hence guiding diagnosis and treatment. Infective endocarditis should be actively looked for in cases of monomicrobial S. anginosus bacteremia, especially if the organism is recovered in multiple blood cultures.

Adult↗

Identification of clinically relevant viridans group streptococci to the species level by PCR.

A PCR assay that allows identification of clinically relevant viridans group streptococci (Streptococcus gordonii, S. mitis, S. mutans, S. oralis, S. salivarius, and S. sanguis) to the species level and identification of milleri group streptococci (S. anginosus, S. constellatus, and S. intermedius) to the group level was developed. This assay was based on specific amplification of internal fragments of genes encoding D-alanine:D-alanine ligases which are species specific and ubiquitous in prokaryotes possessing peptidoglycan. The specificity of this assay was tested on 9 reference strains and 91 characterized clinical isolates. This assay offers a specific and rapid alternative to phenotypic or DNA-DNA hybridization methods for identification of clinically relevant viridans group streptococci.

Alanine↗

Canine urinary tract infections: a comparison of in vitro antimicrobial susceptibility test results and response to oral therapy with ampicillin or with trimethoprim-sulfa.

In vitro susceptibility testing correctly predicted the outcome of ampicillin therapy in all 56 urinary tract infections (UTI) caused by coagulase-positive staphylococci (Staphylococcus aureus and S intermedius), in all 26 UTI caused by Proteus mirabilis, in 38 of 44 UTI caused by Escherichia coli, in 29 of 31 UTI caused by Streptococcus spp, in 8 of 10 UTI caused by Klebsiella pneumoniae, and in 16 of 20 UTI caused by other bacterial species. Thus, 173 of 187 (92.5%) isolates responded to ampicillin therapy in a manner predicted by in vitro susceptibility test results. In vitro susceptibility testing correctly predicted the outcome of therapy with trimethoprim-sulfa in 119 of 138 UTI caused by Escherichia coli, in 33 of 45 UTI caused by Klebsiella pneumoniae, in 38 of 43 UTI caused by Proteus mirabilis, in 21 of 25 UTI caused by Streptococcus spp, in 9 of 11 UTI caused by coagulase-positive staphylococci, and in 19 of 21 UTI caused by other bacterial species. Thus, 239 of 283 (84%) isolates responded to trimethoprim-sulfa therapy in a manner predicted by in vitro susceptibility test results.

Ampicillin↗

Strain-related acid production by oral streptococci.

Acid production, in particular at low pH, is thought to be an important ecological determinant in dental caries. The aim of the present study was to determine the acid producing capability at different pH levels of 47 streptococcal strains, representing 9 species, isolated from human dental plaque. The bacteria were grown until mid log-phase under anaerobic conditions and acid production was measured in a pH-stat system at pH 7.0, 6.0, 5.5 and 5.0. At all pH values, the mean velocity of acid production (V(ap)) by Streptococcus mutans and S. sobrinus was significantly higher (p<0.01; ANOVA) than that of the other oral streptococci, including S. mitis, S. oralis, S. gordonii, S. sanguis, S. intermedius, S. anginosus, S. constellatus, and S. vestibularis. However, the V(ap) of some strains of S. mitis biovar 1 and S. oralis, particularly at pH values of 7.0 and 6.0, exceeded that of some strains of S. mutans. The V(ap) decreased with pH for all strains, but some strains of S. mitis biovar 1 and strains of the mutans streptococci maintained a relatively high rate of acid production. The results suggest that some strains of S. mitis biovar 1 and S. oralis may play an important role in caries development by modifying the environment in dental plaque to become favourable for the succession of aciduric species. The study furthermore emphasises the need for detailed species and biovar identification of oral streptococci and for recognition of the significant physiological differences that occur within single species.

Acids↗

Studies of the subgingival microflora in patients with acquired immunodeficiency syndrome.

Two unique forms of periodontal disease, HIV-gingivitis and HIV-periodontitis, have been described in patients with Acquired Immunodeficiency Syndrome (AIDS). In order to determine the bacterial species associated with periodontitis in AIDS patients, the predominant cultivable microflora was examined in 21 subgingival plaque samples from 11 AIDS patients with periodontitis. The presence of putative periodontal pathogens including Actinobacillus actinomycetemcomitans, Bacteroides intermedius, Porphyromonas gingivalis (formerly B. gingivalis), and Wolinella recta was examined by immunofluorescence in 128 subgingival dental plaque samples from 50 AIDS patients including 32 patients with periodontitis. Of 666 bacterial strains isolated from the 21 subgingival plaque samples, Streptococcus sanguis II was the most frequently recovered species comprising 18.5% of the total number of isolates followed by Lactobacillus acidophilus (12.2%), Porphyromonas gingivalis (12%), Fusobacterium nucleatum (11.4%), Staphylococcus epidermidis (8.7%), Actinomyces naeslundii (7.5%), and Actinomyces viscosus (4.7%). Fusobacterium nucleatum was the most prevalent species and was found in 76% of the sites and 91% of the patients. Enteric species including Enterococcus avium and Enterococcus faecalis, Clostridium clostridiiforme and Clostridium difficle as well as Klebsiella pneumoniae also were recovered. Immunofluorescence assays detected similar carriage rates of A. actinomycetemcomitans, B. intermedius, and P. gingivalis in both gingivitis patients and periodontitis patients, while four times more periodontitis patients demonstrated W. recta. Subgingival yeast was a frequent finding in these AIDS patients, present in 62% of the subjects and 55% of the sites. This study indicates that subgingival plaque in AIDS patients with periodontitis can harbor high proportions of the same periodontal pathogens as are associated with periodontitis in non-HIV infected subjects as well as high proportions of opportunistic pathogens.

Acquired Immunodeficiency Syndrome↗

The establishment of reproducible, complex communities of oral bacteria in the chemostat using defined inocula.

Nine commonly isolated oral bacterial populations were inoculated into a glucose-limited and a glucose-excess (amino acid-limited) chemostat maintained at a constant pH 7.0 and a mean community generation time of 13.9 h. The bacterial populations were Streptococcus mutans ATCC 2-27351, Strep. sanguis NCTC 7865, Strep. mitior EF 186, Actinomyces viscosus WVU 627, Lactobacillus casei AC 413, Neisseria sp. A1078, Veillonella alkalescens ATCC 17745, Bacteroides intermedius T 588 and Fusobacterium nucleatum NCTC 10593. All nine populations became established in the glucose-limited chemostat although Strep. sanguis and Neisseria sp. were present only after a second and third inoculation, respectively. In contrast, even following repeated inoculations, Strep. mutans, B. intermedius and Neisseria sp. could not be maintained under glucose-excess conditions. A more extensive pattern of fermentation products and amino acid catabolism occurred under glucose-limited growth; this simultaneous utilization of mixed substrates also contributed to the higher yields (Y molar glucose) and greater species diversity of these communities. Microscopic and biochemical evidence suggested that cell-to-cell interactions and food chains were occurring among community members. To compare the reproductibility of this system, communities were established on three occasions under glucose-limitation and twice under glucose-excess conditions. The bacterial composition of the steady-state communities and their metabolic behaviour were similar when grown under identical conditions but varied in a consistent manner according to the nutrient responsible for limiting growth. Although a direct simulation of the oral cavity was not attempted, the results show that the chemostat could be used as an environmentally-related model to grow complex but reproducible communities of oral bacteria for long periods from a defined inoculum.

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↗

Bacteriological status of canine milk and septicaemia in neonatal puppies--a retrospective study.

Results of the bacteriological examination of milk samples from 46 bitches were evaluated retrospectively, and correlated with findings of the bacteriological examination of organs from dead, septicaemic puppies in their litters (n = 33). The aim of this study was to investigate, in how many cases of clinical and subclinical mastitis of the bitches, the same bacteria can be detected in their septicaemic neonates. One group of lactating bitches was clinically healthy (group I, n = 38), whereas in eight bitches different puerperal disorders were found (group II). Twenty-five septicaemic puppies were from group I, eight from group II. Out of a broad spectrum of bacteria isolated from the milk of clinically healthy and diseased bitches, only Escherichia coli, Klebsiella pneumoniae and/or ss-haemolytic (haem.) Streptococcus sp. could be isolated from organs of their septicaemic puppies. This was the case in three bitches with mastitis and in one clinically healthy bitch only. Staphylococcus intermedius, although frequently isolated from canine milk, does not seem to be a cause of septicaemia in neonates. It is assumed that in most cases of neonate septicaemia, bacteria from the bitches' milk are not the primary cause.

Animals↗

Nucleotide sequence of the chloramphenicol resistance determinant of the streptococcal plasmid pIP501.

We have sequenced the chloramphenicol resistance determinant (cat) of plasmid pIP501 from Streptococcus agalactiae to investigate its relationship with other cognate cat determinants. Sequence analysis revealed that it exhibits a high degree of similarity with the cat genes of plasmids pC221 and pUB112 from Staphylococcus aureus and pSCS1 from Staphylococcus intermedius. These genes, however, display several differences in their regulatory and coding regions. These results demonstrate that the cat determinant of plasmid pIP501 belongs to the pC221 subgroup of CAT variants.

Amino Acid Sequence↗

[Microbial associations in Trichomonas urethritis].

The results on isolation of associated microflora from 190 patients suffering from Trichomonas urethritis are given. Microorganisms belonged to the species: Mycoplasma hominis. Ureaplasma urealyticum. Streptococcus pyogenes. Escherichia coli. Klebsiella pneumoniae. Proteus mirabilis. Moraxella lacunata. Acinetobacter Calcoaceticus and Candida. Staphylococcus cultures which were isolated from 100% patients belonged to 8 species: Staphylococcus aureus, S. intermedius, S. haemolyticus, S. epidermidis, S. capitis, S. hominis, S. warneri, S. saprophyticus. Most of Staphylococcus cultures had pathogenic features. Structure of Trichomonas vaginalis and interaction between them and associated microflora were studied by electron microscopy.

Animals↗

Bactericidal effects of human neutrophils and sera on selected endodontic pathogenic bacteria in an anaerobic environment.

Phagocytosis by normal human neutrophils and the bactericidal activity of normal human pooled serum were measured under anaerobic conditions on six endodontic pathogenic bacterial species from the genera Actinomyces, Bacteroides, Fusobacterium, Peptostreptococcus and Streptococcus. The results revealed that Actinomyces viscosus was not killed either by human neutrophils or by 20 per cent serum; Peptostreptococcus anaerobius was not killed by either 20 per cent or 95 per cent serum; and Bacteroides intermedius was susceptible to both 20 per cent and 95 per cent serum, but was not killed by neutrophils after exposure for 20 minutes. The other species showed intermediate susceptibility patterns. Such results suggest that oral pathogens vary with regard to the bactericidal effects of these two host defense mechanisms, and that this variation may at least in part explain the relative quantities of microbes recovered in certain oral infections.

Actinomyces↗

Ability of oral bacteria to degrade fibronectin.

The fibronectin-degrading ability of 116, mainly oral, strains was assayed by using plasma-derived fibronectin adsorbed to a polystyrene surface. Ability to degrade fibronectin was revealed in strains of Bacteroides gingivalis, Bacteroides intermedius, Bacteroides loeschii, Staphylococcus aureus, Staphylococcus epidermidis, Peptococcus prevotii, Clostridium sporogenes, and Propionibacterium acnes. The fibronectinolytic activity of subgingival bacteriological samples was found to be related to the presence of B. gingivalis and B. intermedius. In addition, strains of the nonoral Bacteroides species B. asaccharolyticus and B. fragilis showed fibronectin-degrading ability. No such ability was detected in the oral strains tested of Streptococcus, Veillonella, Actinomyces, Lactobacillus, Actinobacillus, Capnocytophaga, Fusobacterium, or Haemophilus species.

Bacteria↗

Bacterial profiles of subgingival plaques in periodontitis.

In this report over 400 subgingival plaque samples taken from over 110 patients were examined microscopically and culturally for 30 bacterial parameters. The patients could be placed into six disease categories based upon clinical criteria. The bacterial profile of each clinical category was generally distinctive of that category. Periodontal patients who had been successfully treated and maintained had plaques that were populated by significantly higher proportions of Streptococcus sanguis, Actinomyces viscosus, A. odontolyticus and S. mutans and significantly lower proportions of B. gingivalis and spirochetes compared to the five untreated disease categories. The spirochetes were the overwhelming microbial type in the plaques of adult periodontitis (AP) patients, averaging about 45% of the microscopic count. The bacteriological results could not distinguish between ADA Type III and IV periodontitis, suggesting that the same type of infection was occurring in an active site in any AP patient. The patients designated as early onset periodontitis (EOP) differed from the other patients by their relative youth and by their significantly higher proportions of Bacteroides gingivalis and/or B. intermedius. Two types of EOP were recognized in which the most diseased variant was characterized by having an average of 49% spirochetes in the plaque. Four localized juvenile periodontitis (LJP) patients were notable in not having detectable A. actinomycetemcomitans. The data indicate that the various types of periodontitis, with the possible exception of LJP are specific anaerobic infections involving spirochetes and to a lesser extent B. gingivalis and B. intermedius.

Actinomyces↗

New combination for the therapy of canine otitis externa. I. Microbiology of otitis externa.

In order to compound a new drug combination against canine otitis externa (OE), 515 dogs affected with OE were subjected to physical examination and microbiological analysis of their ear exudates. OE was erythematous-ceruminous in 83 per cent and suppurative in 17 per cent of the patient material. Erythematous-ceruminous inflammations were characterised by severe pruritus and accumulation of brownish, greasy cerumen in the auditory canal. The yeast Malassezia pachydermatis was isolated from the ears of 76 per cent of the dogs, often in combination with Staphylococcus intermedius bacteria. M pachydermatis showed the most sensitivity, in decreasing order of efficacy, to ketoconazole, econazole, clotrimazole, miconazole and nystatin. S intermedius isolates were most sensitive to amoxycillin-clavulanic acid, enrofloxacin, cephalexin and gentamicin. The microorganism most frequently isolated from dogs with suppurative OE was Pseudomonas aeruginosa; in some cases Proteus, Streptococcus and Pasteurella were also isolated. The P aeruginosa isolates showed the highest sensitivity to gentamicin, polymyxin B and tobramycin.

Aminoglycosides↗