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At least 19 recordsLinked to original sources
PubMed1982

Isolation and characterization of coaggregation-defective mutants of Actinomyces viscosus, Actinomyces naeslundii, and Streptococcus sanguis.

Spontaneously occurring coaggregation-defective (COG-) mutants of oral actinomycetes and streptococci were isolated and used to study interactions between cells of these two kinds of bacteria. COG- mutants of each kind of bacteria were isolated by a simple enrichment scheme. Parent strains were mixed with a coaggregating partner strain, coaggregated cells were removed by low-speed centrifugation, and non-coaggregated cells were recycled by the addition of more partner strain cells. COG- mutants constituted up to 10% of the parent strain cell type in the final enriched cell suspension. Unlike their respective parent strains, COG- mutants of Actinomyces viscosus T14V and Actinomyces naeslundii ATCC 12104, and A. naeslundii I exhibited no lactose-reversible coaggregation with certain oral Streptococcus sanguis strains. However, these COG- mutants were not altered in their coaggregations with another S. sanguis strain, H1, a member of a streptococcal coaggregation group that exhibits only lactose-nonreversible coaggregations with oral actinomycetes. Although all coaggregations between S. sanguis H1 and these actinomycetes appear to be alike, examination of a COG- mutant of S. sanguis H1 revealed that, like its parent, it coaggregated with A. viscosus T14V and its COG- mutants, but unlike its parent, it did not coaggregate with the two A. naeslundii strains or their COG- mutants. Thus, it was concluded that at least two types of surface components are important in mediating coaggregation between S. sanguis H1 and actinomycetes. The COG- mutant of S. sanguis allowed detection of these components, which were indistinguishable in previous studies.

Actinomyces↗
PubMed1993

Synergistic, growth-inhibitory effects of chlorhexidine and copper combinations on Streptococcus mutans, Actinomyces viscosus, and Actinomyces naeslundii.

The purpose of this investigation was to determine whether chlorhexidine combined with copper would exert a synergistic, growth-inhibitory effect against selected oral bacteria. Actinomyces viscosus, Actinomyces naeslundii, and Streptococcus mutans were all susceptible to chlorhexidine individually, with S. mutans displaying the highest sensitivity. Much higher concentrations of copper were needed to achieve growth inhibition of the micro-organisms tested. Determination of minimum inhibitory concentrations (MICs) of chlorhexidine and Cu2+ combinations suggested synergistic activity. Bactericidal kinetics assays confirmed synergism of chlorhexidine and Cu2+ combinations with 1 to 2 log greater decreases in viable cell numbers compared with chlorhexidine alone. Under the constraints of the conditions employed, these data demonstrate the efficacy of chlorhexidine and Cu2+ combinations against the oral bacteria tested. These studies suggest that a chlorhexidine and copper formulation could be useful as a mouthrinse in helping control cariogenic micro-organisms and/or in the treatment of gingivitis.

Actinomyces↗
PubMed1993

Glucose stimulates cAMP accumulation in the oral bacterium Actinomyces viscosus.

Actinomyces viscosus T14V, a Gram-positive bacterium found in the oral cavity, was found to be insensitive to glucose-mediated catabolite repression. Basal levels of beta-galactosidase (18-26 U) were observed at all phases of growth regardless of the culture conditions. Further, beta-galactosidase could not be induced with lactose, or with a known inducer of the enzyme, isopropyl-beta-D-thiogalactoside, or with dibutyryl cAMP. Glucose, on the other hand, stimulated cAMP accumulation in a concentration-dependent manner. Fructose and sucrose mimicked the effects of glucose on cAMP accumulation, whereas galactose, mannose and maltose had lesser stimulatory effects. Other carbon sources, i.e., lactose, alpha-methylglucoside, ribose, xylose and succinate were without effect. Glucose and alpha-methylglucoside were found to stimulate cAMP accumulation in toluene-permeabilized cells, in the presence of the phosphodiesterase inhibitor, theophylline. Glucose did not stimulate cAMP levels in other Gram-positive bacteria including Streptococcus mutans, S. sanguis and S. salivarius but did cause cAMP accumulation in other strains of A. viscosus. The results suggest that glucose effects on cAMP metabolism are independent of the induction of beta-galactosidase as presently defined for Escherichia coli, and that the effects appear to be selective to the A. viscosus bacteria. The results also suggest that glucose stimulates cAMP accumulation via activation of adenylate cyclase.

Actinomyces viscosus↗
PubMed1982

Cell-mediated and humoral immune responses of guinea-pigs to the oral bacterium Actinomyces viscosus.

Actinomyces viscosus is a Gram-positive facultative rod indigenous to most human mouths. The guinea-pig was evaluated as a model for assessing cellular immune responses to a human strain of A. viscosus. Guinea-pigs, immunized with heat-killed A. viscosus cells or a water-soluble extract from A. viscosus (AVS), were tested for delayed-type hypersensitivity responses to A. viscosus 4 weeks after immunization. A week later, the guinea-pigs were terminated. Lymphocytes from peripheral blood, lymph node and spleen were tested for in-vitro blastogenic responses to heat-killed A. viscosus or the water-soluble extract. Plasma from these guinea-pigs were tested for the presence of precipitating antibodies. Positive in-vivo delayed-type hypersensitivity reactions and antibody responses to AVS occurred only in immunized guinea-pigs. A strong in-vitro blastogenic response to AVS or heat-killed A. viscosus occurred in lymphocytes from immunized guinea-pigs, but a weak response was detected in un-immunized animals. These data suggest that the guinea-pig can be used to evaluate different parameters of the immune response to oral bacteria such as A. viscosus.

Actinomyces↗
PubMed1978

Cell-associated levan of Actinomyces viscosus.

Actinomyces viscosus ATCC 15987 was examined for the presence of cell-associated levan by absorption of myeloma proteins with antilevan activity and direct immunofluorescence. Levan was not detectable on the surface of glucose-grown A. viscosus, but after a brief incubation of these cells with 5% sucrose, they were encapsulated with tenaciously adhering levan. The levan layer constituted between 0.02 and 0.03% of the cell dry weight. In contrast, sucrose-grown A. viscosus cells possessed a low level of cell-associated levan, which was only moderately increased by incubation in sucrose and which partially existed as a loose slime rather than a tenacious capsule.

Actinomyces↗
PubMed1978

Degradation of levan by Actinomyces viscosus.

Actinomyces viscosus ATCC 15987 was examined for its ability to hydrolyze its own levan. Washed whole cells and an ammonium sulfate fraction from cell-free culture fluids were shown to possess levan hydrolase activity. Analyses of reaction mixtures by gel filtration and thin-layer chromatography demonstrated that the product of levan hydrolysis was free fructose. The cell-associated and extracellular enzyme preparations also hydrolyzed inulin and the levans synthesized by Aerobacter levanicum and Bacillus subtilis. Growth of A. viscosus in media supplemented with 0.1% A. viscosus levan resulted in a 33-fold increase and a 7-fold increase in the specific activities of the respective extracellular and cell-associated enzymes when compared with those from 55 mM glucose cultures. Growth in the presence of 29.2 mM sucrose resulted in a 28-fold increase and a 5-fold increase in the specific activities of the respective enzymes when compared with those from the glucose cultures. The extracellular enzyme exhibited high activity over a wide pH range, with 87 and 89% of its pH 6.0 optimum activity at pH 5.0 and 7.0, respectively. The cell-associated enzyme also exhibited optimum activity at pH 6.0, but this was decreased to 10 and 20% at pH 5.0 and 7.0, respectively. Analysis for the presence of extracellular levan during growth of A. viscosus in sucrose broths demonstrated that peak levan concentrations occurred during the mid-exponential to late-exponential phase of growth followed by a rapid decline in extracellular levan as a result of levan hydrolase activity.

Actinomyces↗
PubMed1979

The cell wall-associated levansucrase of Actinomyces viscosus.

Actinomyces viscosus produces both a soluble extracellular levansucrase and a cell wall-associated levansucrase. The enzyme from cell walls was solubilized by lysozyme digestion. The soluble extracellular and cell wall-associated forms of the enzyme were compared and appeared to be identical, based on molecular weight estimations, kinetic parameters, and reactions with antisera. The product of both forms of the enzyme was a high molecular weight, branched levan, as shown by its reactivity with myeloma proteins specific for beta(2 leads to 1) and for beta(2 leads to 6) linkages in fructosans. Although levansucrase remained tightly bound to the levan which it synthesized, the enzyme did not bind to exogeneously added levan. Regarding the potential pathogenicity of the levan product, pure levan, produced using purified levansucrase, did weakly activate complement by the alternative pathway. However, the pure levan did not directly cause bone resorption in an in vitro bone resorption assay.

Actinomyces↗
PubMed1988

Distribution of antigenic determinants between Actinomyces viscosus and Actinomyces naeslundii.

A total of 18 monoclonal antibodies was raised against whole cells of Actinomyces viscosus and Actinomyces naeslundii. The monoclonal antibodies were used to determine the cross-reacting patterns among 26 strains of these species. Eleven different antigenic determinants were found. The specificity profiles of the antibodies indicated that the antigenic determinants of A. viscosus and A. naeslundii were arranged in a complicated mosaic. Extensive cross-reactions occurred between A. viscosus strains and strains of "typical" and "atypical" A. naeslundii. However, cross-reactions were rare between the two groups of A. naeslundii. A. viscosus appears to occupy a "middle position" between the two A. naeslundii groups. In addition to their value in seroclassification, some of the monoclonal antibodies were found to be useful in the identification of these species. One monoclonal antibody appeared to be selective for the "typical" A. naeslundii group. A. viscosus and "atypical" A. naeslundii-specific antibodies were also found, though they did not label every strain in their respective clusters. A. viscosus detection might be improved if mixtures of monoclonal antibodies were used.

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PubMed1994

The role of the succinate pathway in sorbitol fermentation by oral Actinomyces viscosus and Actinomyces naeslundii.

The sorbitol fermentation by Actinomyces viscosus and Actinomyces naeslundii was studied with washed sorbitol-grown cells. The fermentation was followed by titration of acids produced at pH 7.0 under anaerobic conditions. Metabolic end-products and intracellular levels of NAD, NADH and glycolytic intermediates during the fermentation were also analyzed. Cell extracts were examined for certain enzyme activities. Bicarbonate was required for acid production from sorbitol and from a mixture of glucose and sorbitol. Malate and fumarate could also support the acid production of A. viscosus. The main end-products were succinate and lactate but not ethanol. Cell extracts showed no activities of alcohol and aldehyde dehydrogenases, but they had activities of malate dehydrogenase and fumarate reductase. In the absence of bicarbonate, malate or fumarate, the intracellular NADH/NAD ratio increased and the levels of 3- and 2-phosphoglycerate and phosphoenolpyruvate decreased. The results indicate that oral sorbitol-fermenting actinomyces lack the ethanol pathway that can contribute to NADH oxidation. To maintain intracellular redox balance during anaerobic sorbitol fermentation, these bacteria can oxidize surplus NADH through a succinate pathway.

Actinomyces↗
PubMed1983

Chemical and immunological comparison of surface fibrils of strains representing six taxonomic groups of Actinomyces viscosus and Actinomyces naeslundii.

Human isolates of Actinomyces viscosus and Actinomyces naeslundii have been divided into six clusters in a numerical taxonomy study. Surface fibrils of strains representing these clusters were isolated and purified. Chemical analyses revealed that the major component of all fibrils was protein and that although differences in percentages of specific amino acid residues were found, the relative proportions of basic, acidic, polar uncharged, and nonpolar amino acids were rather similar among clusters. All of the fibrils except those from strain B236 (cluster 2) either failed to migrate or penetrated only slightly into gels during sodium dodecyl sulfate-polyacrylamide gel electrophoresis, even after boiling, reduction, or alkylation. Immunological studies by electron microscopic examination of fibril-antibody immunocomplexes, whole bacterial cell agglutination, inhibition of hemagglutination, and immunofluorescence by using antifibril antisera and antibodies demonstrated that strains of typical A. naeslundii (cluster 5) have a specific fibril-associated antigen(s) distinct from those of strains of other clusters. Cross-reactions for atypical A. naeslundii (cluster 3) were few. The fibrils from A. viscosus clusters 1, 2, 4, and 6 demonstrated several cross-reactions. By absorbing antifibril antibodies with cross-reactive strains it was possible to obtain cluster-specific antibodies, as determined by whole cell agglutination, only for cluster 5. Absorbed antifibril antisera for both A. naeslundii clusters 3 and 5 were specific by indirect immunofluorescence, whereas anti-cluster 1 fibril antisera cross-reacted only with other A. viscosus cluster representatives. Purification of Actinomyces fibrils by methods used for appendages of other species yields preparations containing common antigens among taxonomic groups. However, absorbing antifibril antisera, gamma globulin, or both has promise for producing cluster-specific reagents useful in identification.

Actinomyces↗
PubMed1975

Differential medium for detecting dental plaque bacteria resembling Actinomyces viscosus and Actinomyces naeslundii.

A medium for detecting colonies of Actinomyces viscosus and Actinomyces naeslundii in dental plaque samples was developed. The medium (CNAC-20) contains 20.0 mug of 3CdSO4-8H2O per ml of Columbia CNA agar base. Laboratory strains of A. viscosus grew on CNAC-20 in characteristic round, white, smooth, opaque colonies. Increasing the cadmium concentration impaired the growth of some A. viscosus strains. Stock strains of A. naeslundii and A. israelii grew in colonies of similar white, opaque morphology. The few strains of other gram-positive plaque bacteria that grew on CNAC-20 had colonies easily distinguished from those of A. viscosus. Most of the bacterial strains freshly isolated from Actinomyces-like colonies on CNAC-20 that had been inoculated with human dental plaque samples were found to have cultural characteristics consistent with previous descriptions of A. viscosus or A. naeslundii. CNAC-20 may facilitate investigations into the relationship of microaerophilic Actinomyces with the etiology of dental diseases.

Actinomyces↗
PubMed1988

Glycogen synthetic and degradative activities by Actinomyces viscosus and Actinomyces naeslundii of root surface caries and noncaries sites.

The relative glycogen synthetic and degradative activities of Actinomyces viscosus and Actinomyces naeslundii, freshly isolated from root surface caries and noncaries sites, were compared. The glycogen synthetic activity was measured by incubating glucose-(or sucrose-)grown resting cells with 100 mM glucose (or sucrose) and U-[14C]-glucose (or U-[14C]-sucrose) on a pH-stat maintained at 5.0, 6.0, and 7.0 for 1 h under anaerobic conditions. For the glycogen degradation assays, after the 1-hour incubation period, the cells were reincubated under similar conditions, but in the absence of external carbon sources. Carbohydrate utilization and total acid formation were also monitored. Both the glucose- and sucrose-grown cells of A. viscosus and A. naeslundii strains originating from root surface caries lesions synthesized approximately twice as much glycogen as the strains of noncaries origin. Although there were significant differences in the rates of glycogen synthesis, the rates of glycogen degradation were essentially the same for the Actinomyces strains from both caries and noncaries sites. However, the time required for glycogen degradation by the strains from caries sites was much longer. This study suggests that the abilities of A. viscosus and A. naeslundii originating from root surface caries lesions to synthesize large amounts of glycogen and to degrade this stored polymer slowly under conditions of starvation, particularly in an acidic environment, may be one of the factors contributing to the cariogenic potential of these organisms in root surface caries.

Actinomyces↗
PubMed1985

Assignment of Actinomyces viscosus and Actinomyces naeslundii strains to numerical taxonomy clusters by immunofluorescence based on antifibril antisera.

A previous observation, using a few representative laboratory strains, that rabbit antisera raised against isolated surface fibrils might be useful in identifying Actinomyces viscosus and Actinomyces naeslundii isolates to their correct numerical taxonomy clusters was reexamined with a large culture collection, including clinical isolates from three different laboratories. Strains were first identified by a slower standard procedure, including agglutination with anti-whole-cell antisera, catalase test, and gas and paper chromatography as needed. Indirect immunofluorescence tests indicated that antisera raised against fibrils of strains representing the three principal taxonomic clusters could separate most strains into their correct clusters. The antisera were particularly successful in separating two taxonomically distinct groups of A. naeslundii, clusters 3 and 5, but could not separate A. viscosus belonging to the principal cluster (no. 1) and minor clusters (no. 2, 4, 6).

Actinomyces↗
PubMed1986

Glycogen synthetic abilities of Actinomyces viscosus and Actinomyces naeslundii freshly isolated from dental plaque over root surface caries lesions and non-carious sites.

Relative glycogen synthetic abilities of resting cells of fresh clinical isolates of Actinomyces viscosus and Actinomyces naeslundii originating from dental plaque samples over root surface caries lesions and non-carious sites were studied under anaerobic conditions at a constant pH of 7.0, with U-(14C)-glucose used as the carbon source. Although the rates of glucose utilization and total acid formation were essentially the same, A. viscosus strains isolated from root surface caries lesions showed glycogen synthetic abilities approximately two to seven times higher than did A. viscosus strains originating from non-carious sites, and also two to four times higher than did A. naeslundii strains originating from both carious and non-carious sites.

Acids↗
PubMed1976

Establishment and distribution of Actinomyces viscosus and Actinomyces naeslundii in the human oral cavity.

The intraoral establishment and proportional distribution of suspected periodontal pathogens Actinomyces viscosus and Actinomyces naeslundii were studied using a recently developed differential plating medium, CNAC-20. Saliva and dental plaque samples were collected from 108 subjects ranging in age from infants to young adults; tongue and buccal mucosa samples were collected from only the adult subjects. Catalase-negative A. naeslundii was isolated from 40% of the predentate infants' and almost all other subjects' saliva samples. It predominated among CNAC-20 isolates in the saliva of subjects of all age groups, in the plaques of young children, and in the adult tongue samples. In contrast, catalase-positive A. viscosus was not isolated from predentate infant samples, and its frequency of isolation increased slowly with age (greater than 50% detection by age 7). A. viscosus was isolated in highest relative proportions from dental plaque and buccal mucosa samples. The two closely related species A. viscosus and A. naeslundii apparently differ in respect to factors determining the host age at which they colonize and their relative intraoral distribution in humans.

Actinomyces↗
PubMed1979

Mannose-contaminating agglutinin for Actinomyces viscosus and Actinomyces naeslundii.

Rapid agglutination of Actinomyces viscosus and Actinomyces naeslundii cells by D-mannose solutions was observed during studies of their attachment to mammalian cells in vitro. The specificity of the agglutination reaction was studied by slide agglutination tests and by measuring the rate of decrease in optical density of bacterial phosphate buffer suspensions caused by the setting of bacterial aggregates. Actinomyces cells were agglutinated by protein-containing mannose solutions of several chemical suppliers. Solutions of sugars other than D-mannose and solutions of mannitol and mannan all failed to agglutinate A. viscsus and A. naeslundii. "Mannose-enhanced" agglutination was impaired by boiling or autoclaving the mannose but was not affected by heating the bacteria, the presence of chloramphenicol, running the assay in the cold, or incorporating any of several commercially purchased sugars in the reaction mixture. During these hapten inhibition experiments, only 6-deoxy-L-talcose-containing extracts of an A. viscosus strain retarded the rate of mannose-enhanced agglutination. Protein-containing fractions of D-mannose mother liquors also agglutinated cells of A. viscosus and A. naeslundii. Other species of oral gram-positive rods were not agglutinated by mannose solutions. Together the data indicate that plant seed-derived D-mannose contains a protein-associated agglutinin for A. viscosus and A. naeslundii which may function via a "lectin-like" selective affinity for the unique cell wall sugar 6-deoxy-L-talose.

Actinomyces↗
PubMed1982

Development of a selective medium for detection and enumeration of Actinomyces viscosus and Actinomyces naeslundii in dental plaque.

A selective medium (CFAT) was developed for the detection and enumeration of Actinomyces viscosus and Actinomyces naeslundii in dental plaque. Neutral acriflavin and potassium tellurite were used in combination with the known selective agents cadmium and fluoride to eliminate most of the competing plaque flora. Composition of CFAT per liter was as follows: Trypticase soy broth (BBL Microbiology Systems), 30 g; glucose, 5 g; agar, 15 g' cadmium sulfate, 13 mg; sodium fluoride, 85 mg; neutral acriflavin, 1.20 mg; potassium tellurite, 2.50 mg; basic fuchsin, 1.25 mg; defibrinated sheep blood, 50 ml. A. viscosus reference strains of human origin grew on CFAT without reduction in numbers under an atmosphere of 90% air-10% CO2. Animal strains of A. viscosus were inhibited at the level of cadmium in CFAT. Two of six A. naeslundii strains did not grow on CFAT. Improved recovery of A. viscosus and A. naeslundii from dental plaque occurred on CFAT, as compared with two other selective media which contained either cadmium sulfate or sodium fluoride, respectively, as selective agents. CFAT was more selective with regard to much of the extraneous gram-positive flora. Bacterionemia, Neisseria, yeasts, and streptococci were virtually eliminated.

Actinomyces↗
PubMed1990

Oxygen-dependent lactate utilization by Actinomyces viscosus and Actinomyces naeslundii.

Actinomyces viscosus and Actinomyces naeslundii were grown in chemostat cultures with glucose or lactate as the sole sources of energy. Growth on lactate was dependent on oxygen. Lactate was oxidised partly to acetic acid. As judged from the low carbon recoveries in acidic degradation products, it is concluded that a substantial part of the lactate was oxidised to carbon dioxide. Assuming that 1 ATP is generated in the transfer of an electron pair to oxygen, YATP values of 10-11 were calculated from the molar growth yield on lactate and product formation. Although all cell 3 test strains oxidised lactate, one strain could not grow on lactate as the sole source of energy. The rate of oxygen uptake with lactate as substrate was higher for cells grown aerobically than of cells grown anaerobically, indicating that lactate oxidation was induced by oxygen. It is reasonable to assume that lactate oxidation is mediated either by lactate dehydrogenase or lactate oxidase, but the pathway in Actinomyces sp. is not yet known. Lactate consumption may be an important trait of Actinomyces sp. that live in an environment limited in energy sources for most of the time.

Actinomyces↗