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Antigenic relationships among oral Actinomyces isolates, Actinomyces naeslundii genospecies 1 and 2, Actinomyces howellii, Actinomyces denticolens, and Actinomyces slackii.

Antigenic relatedness among human strains of oral Actinomyces and similar isolates from cattle has been analyzed by agglutination and immunoblotting. Whole cell agglutination placed A. viscosus serotype II, A. naeslundii serotypes II and III, Actinomyces NV, and strains from numerical taxomonic clusters C1, C2, C3, C4, and C6 into a single group. A. viscosus serotype I cross-reacted weakly with this group. A naeslundii serotype I strains and the cattle isolates Actinomyces denticolens and Actinomyces howellii were distinct. The agglutination results for A. slackii were equivocal. Immunoblots of cell wall extracts developed with non-absorbed sera showed cross-reactivity (23% to 90% antigenic similarity) among all of the strains tested, including A. israelii. The range of antigenic similarities among the group which included strains of A. viscosus serotype II, the A. naeslundii serotypes, and clusters C1, C2, C3, C4, and C6 was from 39% to 89%. Immunoblotting showed that A. howellii and A. denticolens were between 39% and 72% similar to A. naeslundii and A. viscosus. Absorption of antisera with A. israelii cell walls removed antibodies recognizing antigens common to Actinomyces and made the sera more specific. Immunoblotting with absorbed sera supported the grouping and separation of strains shown by agglutination. In some cases, serotypes could be included into a specific taxonomic cluster. A. naeslundii serotype II and Actinomyces NV most closely resembled cluster C1 strains, and A. naeslundii serotype III resembled cluster C1 strains, and A. naeslundii serotype I and A. viscosus serotype I were included into clusters C5 and C7, respectively. The results support a recent proposal that strains of A. viscosus serotype II, A. naeslundii serotypes II and III, and Actinomyces NV be included into A. naeslundii genospecies 2, that A. naeslundii serotype I should be designated A. naeslundii genospecies 1, and that A. viscosus serotype I should be retained distinct from A. naeslundii, as A. viscosus.

Absorption↗

Actinomyces georgiae sp. nov., Actinomyces gerencseriae sp. nov., designation of two genospecies of Actinomyces naeslundii, and inclusion of A. naeslundii serotypes II and III and Actinomyces viscosus serotype II in A. naeslundii genospecies 2.

DNAs of type strains and representative members of Actinomyces groups from the human periodontal flora and from other habitats were compared by using the S1 nuclease procedure to determine their genetic relatedness. One rather common group from the human periodontal flora, previously called "Actinomyces D08," is phenotypically distinct from, and genetically unrelated to, previously described species. We propose the name of Actinomyces georgiae for this organism; the type strain is strain ATCC 49285. Another common group from the human periodontal flora is Actinomyces israelii serotype II, which was found genetically distinct from the type strain of A. israelii (serotype I) and from other previously described species of Actinomyces. We propose the name Actinomyces gerencseriae for this organism; the type strain is strain ATCC 23860. A. naeslundii serotype I strains were distinct from the other strains studied. A separate genospecies which included strains of A. naeslundii serotypes II and III and A. viscosus serotype II was delineated. Strains of Actinomyces serotype WVA 963 constitute an additional distinct genospecies. Because there are no reliable phenotypic tests, other than serological analyses, to differentiate Actinomyces serotype WVA 963 and the two genospecies of A. naeslundii, no taxonomic changes are proposed for these three genospecies.

Actinomyces↗

Assignment of Actinomyces pyogenes-like (CDC coryneform group E) bacteria to the genus Actinomyces as Actinomyces radingae sp. nov. and Actinomyces turicensis sp. nov.

In a previous study the authors reported the characterization of some facultatively anaerobic, Gram-positive, non-sporeforming rods which were found in mixed cultures from various infectious processes, including patients with otitis, empyema, perianal abscesses and decubitus ulcers. Phenotypically these organisms closely resembled Actinomyces pyogenes although their precise taxonomic position remained unknown. In the present investigation the authors have determined the 16S rRNA gene sequences of some representative strains of the Actinomyces pyogenes-like bacteria and report the results of a comparative sequence analysis. On the basis of the results of the present and earlier findings two new Actinomyces species, Actinomyces radingae sp. nov. and Actinomyces turicensis sp. nov. are proposed. The type strains are DSM 9169T and DSM 9168T, respectively.

Actinomyces↗

Assignment of human-derived CDC group 1 coryneform bacteria and CDC group 1-like coryneform bacteria to the genus Actinomyces as Actinomyces neuii subsp. neuii sp. nov., subsp. nov., and Actinomyces neuii subsp. anitratus subsp. nov.

Almost the entire 16S rRNA gene sequences of some strains of CDC group 1 and group 1-like coryneform bacteria, isolated from human sources, were determined. Comparative analysis of the rRNA sequence data revealed that both groups of coryneforms belong to the genus Actinomyces. On the basis of the present molecular findings and previous biochemical studies, we propose a new Actinomyces species, Actinomyces neuii sp. nov., containing Actinomyces neuii subsp. neuii subsp. nov. for CDC group 1 coryneform bacteria and Actinomyces neuii subsp. anitratus subsp. nov. for CDC group 1-like coryneform bacteria.

Actinomyces↗

Comparative histopathology of lesions produced by Actinomyces israelii, Actinomyces naeslundii, and Actinomyces viscosus in mice.

The histopathologic features of experimental actinomycotic lesions produced in mice by Actinomyces israelii, Actinomyces naeslundii, and Actinomyces viscosus were examined. In lesions caused by A israelii the outer edge of the bacterial granule exhibited an eosinophilic fringe with no evidence of penetration of polymorphonuclear leukocytes (PMNs) into the bacterial granule. Chronic lesions after 6 weeks contained lobulated advancing fronts as well as areas of resolution showing heavy penetration by phagocytic cells. The number of macrophages and plasma cells in these lesions increased with time. In contrast, lesions caused by A viscosus and A naeslundii showed cellular evidence of resolution during the early stages of the infection (3-6 weeks). The bacterial core was readily penetrated and fragmented by PMNs in early A viscosus lesions. In lesions caused by A naeslundii there was less penetration of the central core by PMNs, and the bacterial granule tended to retain its structural integrity. Elongated crystals of hyaloid material appeared in lesions caused by all species. These protein-rich bodies appeared to be associated with resolving areas of the lesions. The observed contrast in the histopathologic appearance of actinomycotic lesions caused by A israelii, A naeslundii, and A viscosus is suggestive of important differences in the immune response of the host to infections caused by these three species.

Actinomyces↗

Sialidase-enhanced lectin-like mechanism for Actinomyces viscosus and Actinomyces naeslundii hemagglutination.

Laboratory strains representing six numerical taxonomy clusters and fresh isolates of human Actinomyces viscosus and Actinomyces naeslundii were studied by standard flocculation slide tests for the ability to hemagglutinate erythrocytes (RBC) from various animal species. Human AB and horse RBC were agglutinated more frequently and rapidly than others; guinea pig RBC were agglutinated by only a few strains. Human AB RBC were selected for studies of hemagglutination mechanisms. Treatment of RBC with clostridial neuraminidase (NTRBC) greatly enhanced hemagglutination for almost all strains. In hapten inhibition experiments in which various concentrations of sugars were used, beta-galactosides were the most effective inhibitors of hemagglutination for both RBC and NTRBC; inhibition of NTRBC agglutination required higher concentrations. Soybean lectin agglutinated both RBC and NTRBC but not Actinomyces cells. NTRBC agglutinated at a 125-fold-lower concentration. Hemagglutination was sensitive to ethylenediaminetetraacetate for one strain tested. Hemagglutination reactions were reversible by addition of beta-galactosides. The ability of Actinomyces strains to "prime" RBC for hemagglutination by removing sialic acid to expose more penultimate beta-galactoside sites was studied by recycling Actinomyces-agglutinated RBC which were dispersed with a lactose solution and washed free of bacteria (primed RBC). Priming in this manner augmented subsequent hemagglutination by indicator Actinomyces strains and made the RBC more sensitive to agglutination by soybean lectin. The priming ability of Actinomyces strains generally correlated with the amount of sialic acid removed from primed RBC. Strains representing the numerical taxonomy clusters differed in both their hemagglutinating and priming activities. Cluster 5 strains (typical A. naeslundii) were good agglutinators of RBC, NTRBC, and primed RBC but were poor primers. Cluster 3 strains (atypical A. naeslundii) were the weakest hemagglutinators but could prime RBC adequately for subsequent agglutination by other strains. Together, these data indicate that Actinomyces hemagglutination proceeds via a two-step mechanism: (i) neuraminidase removal of terminal sialic acid and (ii) lectin-like binding to exposed beta-galactoside-associated sites on the RBC. Strains differ in the extent to which they can perform the two functions, and this specificity may relate to their taxonomic classification.

Actinomyces↗

Characterization of some Actinomyces-like isolates from human clinical specimens: reclassification of Actinomyces suis (Soltys and Spratling) as Actinobaculum suis comb. nov. and description of Actinobaculum schaalii sp. nov.

Five strains of a hitherto unknown Actinomyces-like bacterium were isolated from human clinical sources, including blood cultures. Biochemical and chemotaxonomic characterization indicated that the strains were distinct from previously described Actinomyces and Arcanobacterium species. A comparative 16S rRNA gene sequence analysis demonstrated that the undescribed strains constitute a new subline within the Actinomyces-Arcanobacterium species complex. The closest known relative of the isolates was found to be Actinomyces suis, although a 16S rRNA sequence divergence value of approximately 6% clearly demonstrated that the unknown bacterium represents a distinct species. Based on the results of the present and earlier phylogenetic investigations, it is proposed that Actinomyces suis should be reclassified in a new genus, the genus Actinobaculum, as Actinobaculum suis comb. nov. In addition, a new species, Actinobaculum schaalii, is proposed for the Actinomyces-like bacterium from human sources. The type strain of Actinobaculum schaalii is CCUG 27420.

Actinomyces↗

Characterization of Actinomyces isolates from samples from the human urogenital tract: description of Actinomyces urogenitalis sp. nov.

Three strains of a previously undescribed Actinomyces-like bacterium were isolated from human clinical sources (urine, urethra and vaginal secretion). Biochemical testing and PAGE analysis of whole-cell proteins indicated that the strains were phenotypically homogeneous and distinct from previously described Actinomyces and Arcanobacterium species. Comparative 16S rRNA gene sequencing studies showed the bacterium to be a hitherto unknown subline within a group of Actinomyces species which includes Actinomyces bovis, the type species of the genus. Based on phylogenetic and phenotypic evidence it is proposed that the unknown bacterium from humans be classified as Actinomyces urogenitalis sp. nov. The type strain of Actinomyces urogenitalis is CCUG 38702T (= CIP 106421T).

Actinomyces↗

Characterization of Actinomyces isolates from infected root canals of teeth: description of Actinomyces radicidentis sp. nov.

Two strains of a previously undescribed Actinomyces-like bacterium were recovered in pure culture from infected root canals of teeth. Analysis by biochemical testing and polyacrylamide gel electrophoresis of whole-cell proteins indicated that the strains closely resembled each other phenotypically but were distinct from previously described Actinomyces and Arcanobacterium species. Comparative 16S rRNA gene-sequencing studies showed the bacterium to be a hitherto unknown subline within a group of Actinomyces species which includes Actinomyces bovis, the type species of the genus. Based on phylogenetic and phenotypic evidence, we propose that the unknown bacterium isolated from human clinical specimens be classified as Actinomyces radicidentis sp. nov. The type strain of Actinomyces radicidentis is CCUG 36733.

Actinomyces↗

Phylogenetic evidence for the transfer of Eubacterium suis to the genus Actinomyces as Actinomyces suis comb. nov.

The 16S rRNA primary structures of Eubacterium suis DSM 20639T (T = type strain) and Bifidobacterium bifidum DSM 20456T were determined by sequencing in vitro amplified rDNA. Sequence comparisons indicated that B. bifidum is moderately related to representatives of the genera Actinomyces and Mobiluncus. The closest relative of E. suis is Actinomyces pyogenes. E. suis and A. pyogenes are more closely related phylogenetically to one another than to the other Actinomyces species that have been investigated by using comparative 16S rRNA analysis. Therefore, we propose that E. suis should be transferred to the genus Actinomyces as Actinomyces suis comb. nov.

Actinomyces↗

Molecular taxonomic studies of Actinomyces-like bacteria isolated from purulent lesions in pigs and description of Actinomyces hyovaginalis sp. nov.

The 16S rRNA gene sequence of some Actinomyces-like bacteria isolated from purulent lesions in pigs was determined. A comparative analysis of the rRNA sequence data revealed that the bacteria are members of the genus Actinomyces, but are phylogenetically distinct from Actinomyces suis. On the basis of our findings and the results of previous phenotypic studies it is formally proposed that the bacteria from pigs should be designated a new species, Actinomyces hyovaginalis.

Actinomyces↗

Partial 16S rRNA primary structure of five Actinomyces species: phylogenetic implications and development of an Actinomyces israelii-specific oligonucleotide probe.

The intra- and intergeneric relationships of the genus Actinomyces were determined by comparing long 16S rRNA sequences, generated by reverse transcriptase. All species formed a phylogenetically coherent cluster in which Actinomyces bovis, A. viscosus, A. naeslundii, A. odontolyticus and A. israelii constituted genetically well defined species. A. israelii DSM 43322 (serotype 2) was not closely related to three other strains of this species (serotype 1) and, as judged from phylogenetic distances, could be accommodated within A. naeslundii, or represent a new species. In contrast to previous findings, members of the genus Actinomyces appear to be related to Bifidobacterium bifidum. Sequence information was used to develop an oligonucleotide probe for the A. israelii serotype 1 strains, which did not react with the serotype 2 strain or with rRNA from strains of eight Actinomyces species.

Actinomyces↗

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↗

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↗

New medium for isolation of Actinomyces viscosus and Actinomyces naeslundii from dental plaque.

Metronidazole (10 microgram/ml) and cadmium sulfate (20 microgram/ml) were added to a gelatin-based medium to select for microaerophilic Actinomyces species from dental plaque samples. The new medium (GMC), when incubated anaerobically, allowed 98% recovery of seven pure cultures of Actinomyces viscosus and 73% recovery of eight pure cultures of Actinomyces naeslundii, while suppressing 76% of the total count of other organisms in dental plaque samples. In 203 plaque samples, recoveries of A. viscosus and A. naeslundii on GMC and another selective medium for oral Actinomyces (CNAC-20) were compared. Recovery of A. viscosus was comparable on the two media. Recovery of A. naeslundii was significantly higher on GMC than CNAC-20 (P is less than 0.001), and GMC allowed a more characteristic cell morphology of both organisms. GMC medium appears to be useful for the isolation and presumptive identification of A. viscosus and A. naeslundii from dental plaque.

Actinomyces↗

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↗