Strains Of Actinomyces Research Articles

Immunoglobulins in Milk from Cows Immunized with Oral Strains of Actinomyces, Prevotella, Porphyromonas, and Fusobacterium

Immunization of pregnant cows with bacteria leads to the presence of high concentrations of specific antibodies in colostrum and milk. A total of 14 cows was immunized with single strains of heat-killed oral bacteria or pools of strains of Actinomyces, Porphyromonas, Prevotella, and Fusobacterium. Two cows were treated with adjuvant alone. The mean percentages of IgG1, IgG2, IgM, and IgA in all of the milks were 83.8, 3.8, 9.3, and 3.1, respectively. ELISA and whole cell agglutination assays demonstrated high titers in the milks from the cows immunized with either individual strains or the bacterial pools. The highest titers determined by ELISA belonged to the IgG1 isotype and in several milks were 64-fold greater than titers in milk from cows treated with adjuvant alone. The concentrations of all antibodies and the titers determined by ELISA and whole cell agglutination assays markedly decreased from the first to the sixth milkings. The functional specificity of the antibodies was demonstrated by agglutination tests against a wide range of bacteria including members of Actinomyces, Fusobacterium, Porphyromonas, Prevotella, Streptococcus, Eubacterium, Propionibacterium, Peptostreptococcus, Bacteroides, Actinobacillus, Haemophilus, Capnocytophaga, and Wolinella. Minimal cross-reactions with bacteria in other genera were observed with all of the milks. High-titer milk preparations have been obtained from immunized cows, and the capacity of the bovine antibodies to agglutinate target bacteria indicates their potential usefulness in oral passive immunization studies.

Salivary receptors for GalNAc beta-sensitive adherence of Actinomyces spp.: evidence for heterogeneous GalNAc beta and proline-rich protein receptor properties.

The receptors for GalNAc beta 1-3Gal alpha Oethyl (GalNAc beta)-sensitive adherence of Actinomyces strains to salivary pellicles were investigated. Parotid and submaxillary saliva from one individual was size fractionated and utilized in hydroxyapatite adherence assays with Actinomyces naeslundii 12104 and A. viscosus 19246 and LY7 with and without GalNAc beta. Three parotid salivary fractions, the high-molecular-weight, acidic proline-rich protein (PRP), and statherin fractions, promote GalNAc beta-sensitive adherence of strain 12104, whereas only the high-molecular-weight fraction of submaxillary saliva promotes such adherence. In contrast, strain LY7, possessing a variant GalNAc beta specificity, shows GalNAc beta-sensitive adherence to the leading and trailing regions of the submaxillary PRP fractions but less distinct adherence to the parotid and submaxillary high-molecular-weight fractions. In addition, the PRP and statherin fractions promote adherence of strains LY7 and 19246 that is not inhibited by GalNAc beta. However, whereas strain LY7 binds more strongly to the PRP fraction than to the statherin fraction, strain 19246 binds preferentially to the statherin fractions of parotid and submaxillary saliva. These salivary protein fractions were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunostained to detect glycosylated proteins. The different salivary receptor properties are paralleled by different glycosylation patterns. The variable GalNAc beta specificities may have evolved to match different salivary glycosylation patterns, and PRP and statherin binding properties seem to be heterogeneous among the Actinomyces strains.

Nonspecific Antibacterial Factors in Milk from Cows Immunized with Human Oral Bacterial Pathogens

Both the immunoglobulins and non-specific antibacterial factors in milk from cows immunized with pathogenic oral bacteria have the potential to influence the oral microflora during passive immunization studies. The first six milks after calving were collected from 2 cows immunized with adjuvant and from 14 cows immunized with adjuvant and heat-killed strains of periodontopathic Actinomyces, Porphyromonas, Prevotella, and Fusobacterium. Analysis of the products from the first to the sixth milks revealed that the protein and lysozyme content decreased approximately 66 and 72%, respectively; the mean specific activity of the enzyme remained relatively constant. In contrast, the mean lactoperoxidase activity increased 2.3-fold in the second milking and increased further in the fourth and sixth milkings. The mean iron-binding activity increased 1.2-fold from the first to the second milkings and then decreased 3.6-fold through the sixth milking. Cows immunized with adjuvant alone showed similar responses. Per unit volume, the milk contained approximately 150 times less lysozyme than whole human saliva obtained from six subjects but higher concentrations of lactoperoxidase and iron-binding components. Purified bovine nonspecific factors prevented the growth of the bacteria used for immunization when bacteria were tested at concentrations similar to those found in saliva and milk. Because bovine nonspecific antibacterial factors could influence both the pathogenic target bacteria and the indigenous microflora in oral passive immunization studies with bovine immunoglobulins, the presence of these proteins should be considered.

Conservation of an Actinomyces viscosus T14V type 1 fimbrial subunit homolog among divergent groups of Actinomyces spp.

The type 1 fimbrial subunit gene of the human Actinomyces viscosus T14V was used as a DNA probe in Southern analyses to detect related DNA sequences in 16 of 30 strains of Actinomyces spp. under conditions of high stringency. The organisms with homology to the DNA probe included two human and six nonhuman A. viscosus, three human and three nonhuman A. naeslundii, and two A. bovis isolates. Homologous DNA sequences were not detected in strains of A. odontolyticus and A. israelii examined in this study. Northern (RNA) blot analysis revealed expression of a transcript from each of the A. viscosus and A. naeslundii strains and from one A. bovis strain that was comparable in size to that detected from A. viscosus T14V. Cell surface fimbriae were observed on a majority of the strains that expressed the transcript. Various degrees of cross-immunoreactivities between these strains and antibodies specific for type 1 fimbriae of A. viscosus T14V were also observed by colony immunoassay. Thus, the data clearly demonstrate the existence in, and expression by, divergent Actinomyces groups of genomic sequences that are closely related to the type 1 fimbriae of A. viscosus T14V.

A Comparison of the Adhesion Profiles and Cell Surface Characteristics ofStreptococcus mitiswith those of Other Members of theStreptococcus sanguisGroup

A collection of 12 strains of Streptococcus mitis were tested for their in vitro adhesion profiles and compared to those of other strains within the Streptococcus sanguis group (SSG). The S. mitis isolates from saliva and the strains isolated from plaque gave significantly greater cell surface hydrophobicities ( P < 0.01) than isolates from other sites. The saliva isolates were generally not aggregated by saliva wherea s S. sanguis I and II strains gave aggregation values over a wide range. Strains from all species gave a wide range of values for adhesion to saliva-coated hydroxyapatite (SHA), but isolates from dental plaque adhered significantly ( P < 0.01) better to SHA than other isolates. The adhesion to SHA was not related to cell surface hydrophobicity or salivary aggregation. Strain pairs were allowed to interact with each other in the SHA adhesion assay. Homologous pairings usually gave reduced adhesion as did some non-homologous pairings, possibly as the result of occupying similar salivary receptors. Occasionally non-homologous pairings increased the adhesion of one member suggesting that certain SSG strains can modify the salivary pellicle to facilitate the adhesion of other SSG strains. Most strains were poor adherers to buccal epithelial cells (BEC), although S. sanguis I strains were more adhesive ( P < 0.025) than other species/biotypes. Serological tests on SSG strains for surface antigens showed 12 per cent reactivity with antisera to lipoteichoic acid and 40 per cent reactivity with antisera to protein P1; in neither case was there a correlation with adhesion to SHA or BEC or aggregation by saliva. Strains of S. mitis were more likely to coaggregate with veillonella than with actinomyces strains, whereas strains of S. sanguis tended to coaggregate with representatives of both genera. Keywords: Adhesion; Saliva; Streptococcus mitis ; Streptococcus sanguis ; Surface antigens; Coaggregation; Hydrophobicity.

Aggregation of oral bacteria by human salivary mucins in comparison to salivary and gastric mucins of animal origin

Seventeen strains of oral bacteria of the genera Actinomyces (5), Bacteroides (3), and Streptococcus (9) were tested for aggregation by the human whole salivary mucin fraction (HWSM) in comparison to three types of animal mucin preparations from submandibular glands of cow (BSM) and sheep (OSM), and from the stomach of pig (PGM). Considerable variation was seen with respect to the rate and titer of aggregation induced by these mucins. The aggregating activity of HWSM varied widely among the different bacterial strains. The Bacteroides group showed hardly any induced aggregation, whereas the final aggregation titers varied for S. sanguis (3 strains) between 12 and 48, for S. oralis (3 strains) between 6 and 48, for the S. mutans group (3 strains) between 6 and 96, and for the five Actinomyces strains even between 6 and 192. For a particular strain, similar differences in titer were seen between the four mucins. For a human salivary mucin (MG-2) it has been described that sialic acid in the sequence NeuAc (alpha 2,3)Gal(beta 1,3)GalNac- was specifically involved in the interaction with S. sanguis strains, in contrast to S. rattus BHT. Our results, however, indicate that this sugar sequence is not a prerequisite for the aggregation of S. sanguis, as animal mucins, devoid of this structure, were equally well or even better capable of inducing aggregation. On the other hand, desialization of BSM and OSM largely abolished their aggregating capability towards S. rattus BHT. Moreover, it was found that BSM and OSM, which are comparable with respect to their major oligosaccharide structure, show considerable differences in aggregating activity towards the same bacterial strain.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of the ATB 32 A system for identification of anaerobic bacteria isolated from clinical specimens

A new miniaturized 4-h method for the identification of anaerobic bacteria, ATB 32 A (API System SA, Montalieu Vercieu, France), was evaluated against conventional methods of identification. The evaluation was done by using 260 recent clinical isolates and 21 reference strains of anaerobic bacteria. All reference strains were correctly identified and did not figure in the detailed analysis. Of the 140 gram-negative bacilli, 90.6% of Bacteroides spp. and 95.5% of Fusobacterium spp. were correctly identified to the species level, with an additional 8.4% of the Bacteroides spp. being identified to the genus level. Clostridia were correctly identified in 85.9% of cases, with an additional 9.9% being identified to the genus level. Peptostreptococci were correctly identified in 91.6% of cases. The 4 strains of Actinomyces spp. were all identified correctly, as were 10 of the 11 strains of Propionibacterium spp. A total of 3.1% of strains were not identified by ATB 32 A, while for 1.9% of strains, completely false identifications were obtained. Estimation of the individual preformed enzyme results may pose problems, although these decrease with familiarity with the system. With certain enzyme profiles, additional testing was necessary to arrive at an identification; however, there was no requirement for gas-liquid chromatography. If certain additions are made to the data base and the difficulties of determination of organisms to the species level among the non-Bacteroides fragilis (sensu stricto) members of the B. fragilis group can be reduced, this system holds promise as a reliable standardized alternative for the identification of anaerobic bacteria in clinical laboratories.

Tetrameric manganese superoxide dismutases from anaerobic Actinomyces

Superoxide dismutase was isolated from each of the anaerobically grown organisms Actinomyces naeslundii, Actinomyces strain E1S.25D, and Actinomyces odontolyticus. The enzymes were 100,000–110,000 mol wt acidic proteins (p I 4.3–4.6) and contained Mn and Zn, but no detectable Fe. The Mn and Zn content varied with the enzyme source. A. naeslundii superoxide dismutase, specific activity 2200 U/mg, contained 2.3 g atoms Mn and 1.4 g atoms Zn per mole tetramer whereas A. odontolyticus SOD, specific activity 700 U/ mg, contained 1.4 g atoms Mn and 1.8 g atoms Zn per mole tetramer. Actinomyces strain E1S.25D, specific activity 1300 U/mg, contained 1.8 g atoms Mn and 1.2 g atoms Zn per mole tetramer. The amino acid compositions of the enzymes were comparable except for arginine, lysine, and tryptophan content. The enzymatic activity of each enzyme was stable in 5 m m H 2O 2 at 23 °C for 2 h. The enzymes were only modestly inhibited by 20 m m NaN 3. The enzymatic activity was increased at low ionic strength but was markedly decreased at increased ionic strength with each salt tested except sodium perchlorate, which caused marked inhibition even at low ionic strength. Polyclonal antibodies to A. naeslundii and Actinomyces strain E1S.25D precipitated and inactivated their respective antigens whereas the precipitated A. odontolyticus superoxide dismutase-antibody complex retained virtually full catalytic activity. Immunological studies revealed that the native A. naeslundii and Actinomyces strain E1S.25D MnSODs share common epitopes and cross-reacted with precipitin lines of complete identity in Ouchterlony double diffusion gels. Antibody to the A. odontolyticus enzyme displayed only partial cross-reactivity with superoxide dismutase from the two other Actinomyces. Western blotting of the denatured antigens revealed reactivities of the antibodies that differed only slightly from the results of the Ouchterlony gels.

Cellular coaggregation of oral Streptococcus milleri with actinomyces

Oral isolates of Streptococcus milleri were examined for their ability to coaggregate with actinomyces. Of the 68 S. milleri strains tested, including 3 reference strains, 40 strains coaggregated with Actinomyces naeslundii WVU45 (actinomyces coaggregation group B) and 36 strains coaggregated with Actinomyces viscosus T14V (actinomyces coaggregation group A). All S. milleri strains of serotypes b (4 strains), e (2 strains), and f (24 strains) coaggregated with both of the actinomyces. The coaggregation reactions between the S. milleri cells and A. naeslundii WVU45 cells were optimal at about pH 7.0 and were Ca2+ or Mg2+ dependent, but they were not inhibited by the presence of simple sugars or amino sugars, including lactose (up to 0.5 M). Treatment of the S. milleri cells with heat (100 degrees C, 3 min) or proteases (trypsin, 1.0 mg/ml; pronase, 0.25 mg/ml; 37 degrees C; 3 h) and of the actinomyces cells with periodate (0.01 M, 4 degrees C, 16 h) destroyed their coaggregating abilities. The coaggregations between cells of the S. milleri strains, we well as cells of the Streptococcus sanguis H1 (reference strain for streptococcus coaggregation group 2) and the actinomyces strains (WVU45 and T14V), were inhibited by AFH1 (a carbohydrate receptor on T14V cells for a lectin on H1 cells). These interactions were also inhibited by anti-AFH1 immunoglobulin G (IgG) and by anti-b, anti-e, and anti-f S. milleri IgG or anti-f IgG Fab fragments. These results suggest that S. milleri, at least strains of serotypes b, e, and f, belongs to streptococcus coaggregation group 2.

Aggregation of 27 oral bacteria by human whole saliva. Influence of culture medium, calcium, and bacterial cell concentration, and interference by autoaggregation.

Twenty-seven oral strains of the genera Actinomyces (5), Bacteroides (3), and Streptococcus (19) were tested for aggregation by human whole saliva, as well as the effect of culture medium, Ca-ions, and bacteria concentration thereupon. Of the media tested, GF-broth gave rise to less interference by autoaggregation or higher aggregation titers than BHI and TSB, and was used throughout this study. In most cases, Ca-ions (1 mM) only enhanced the rate of induced aggregation, whereas raising the bacteria concentration increased the rate of both induced- and autoaggregation. The final titers, ranging from 1-64, were hardly affected by these parameters, except those of S. rattus HG 59 and S. mutans HG 199, which were respectively increased and decreased by Ca-ions. Saliva-induced aggregation was observed for 21 strains of A. viscosus, A. naeslundii, A. israelii, B. gingivalis, B. intermedius, S. cricetus, S. mutans, S. rattus, S. sanguis, and S. sobrinus, mostly within 15 min to 3 h. Seventeen of these strains also showed autoaggregation, usually well after the onset of induced aggregation. Any potential induced aggregation of B. gingivalis HG 91 was always masked by autoaggregation, as well as that of the S. mutans strains under a particular set of conditions. The aggregation rate and titer varied considerably in a mutually unrelated and strain-dependent way. These microtiterplate data were matched by the 5 spectrophotometric patterns observed for saliva-bacterial interaction, which moreover, gave the better differentiation between induced and autoaggregation. In conclusion, most strains tested can show rapid saliva-induced aggregation in a strain-dependent way, yet strongly affected by the experimental conditions and interference from autoaggregation.

Immunochemical studies on pneumococcal type 37 capsular polysaccharide

Pneumococcal type 37 capsular polysaccharide was obtained free of contaminants by affinity chromatography on Con-A, wheat germ agglutinin, Maclura pomifera lectin and HOPC-8 mouse myeloma protein affinity columns. The immunochemical reactivity of native and periodate oxidized borohydride reduced type 37 polysaccharide antigen with polyclonal rabbit and monoclonal mouse anti-Pn37 hybridoma antibodies was studied by quantitative precipitation. Quantitative hapten inhibition studies, employing the isomeric series of α- and β-(1 → 2), (1 → 3), (1 → 4) and (1 → 6)-linked glucobioses as competitive inhibitors of antibody precipitation establish a specificity for anti-Pn37 antibody directed at least in part, against the Glc β(1 → 2) Glc (sophorosyl) unit. A high mol. wt, d-glucose containing polysaccharide antigen, cross-reactive with rabbit anti-Pn37 is reported which was found to occur in the culture medium of 7 of 19 strains of Actinomyces examined.

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.

Use of lytic bacteriophage for Actinomyces viscosus T14V as a probe for cell surface components mediating intergeneric coaggregation.

A lytic bacteriophage for Actinomyces viscosus T14V (the reference strain for actinomyces coaggregation group A) was isolated from raw sewage. This phage, designated BF307, also lysed the T14V-derived nonfimbriated mutant PK455-2 as well as A. viscosus MG-1 and T14AV but not the other serotype 2 or serotype 1 strains of this species that were tested or any of nine Actinomyces naeslundii isolates. Phages BF307 belonged to Bradley morphological group C and was similar in appearance to the A. viscosus MG-1 phages Av-1 and Av-3, which do not productively infect A. viscosus T14V. A. viscosus MG-1 mutants selected for resistance to phage BF307, Av-3, or CT7 (a human dental plaque isolate with the same host range as BF307) were coresistant to the other two phages but sensitive to Av-1. These results indicate that the receptors on A. viscosus MG-1 for phages BF307, Av-3, and CT7 are identical or share a common precursor and that the receptor for phage Av-1 is distinct. Comparison of the genomes of BF307, Av-3, and CT7 revealed that their DNAs were similar in size but distinguishable by restriction analysis. Two altered coaggregation phenotypes were identified among the phage BF307-resistant mutants of strains MG-1, T14V, T14AV, and PK455-2. Class I mutants had lost the ability to interact with coaggregation group 1 streptococci, and class II mutants did not coaggregate with either group 1 or group 2 streptococci. These results are consistent with the proposal that the phage BF307 receptor on these A. viscosus strains is related to one of the structures that mediates coaggregation with oral streptococci. A model to delineate the various coaggregation mediators on the surface of actinomyces coaggregation group A cells is presented, and the use of these phages to probe surface components of human oral actinomyces strains is discussed.

Intergeneric bacterial coaggregations involving mutans streptococci and oral actinomyces.

Mutans streptococci (MS) representing eight different serotypes were tested for their ability to coaggregate in vitro with oral actinomyces and other streptococcal species. Of the mutans streptococci tested, only strains of S. cricetus (formerly S. mutans serotype a) displayed pronounced coaggregations and only with certain strains of actinomyces. S. cricetus coaggregated, by lactose nonreversible mechanisms, with serotype 4 Actinomyces naeslundii WVU963 and WVU924 and with serotype 2 Actinomyces odontolyticus WVU758. The first pair was disaggregated by protein denaturants (e.g., sodium dodecyl sulfate and urea) and EDTA. This coaggregation was inhibited when the streptococcal, but not the actinomyces, partner was pretreated with either heat or protease, suggesting the presence of a protein mediator on only the streptococcal cell surface. The S. cricetus-A. odontolyticus coaggregation appeared to involve protein components on each cell, as shown by the lack of coaggregation after pretreatment of either cell type with heat or proteases. This coaggregation was also reversed by sodium dodecyl sulfate and urea, as well as by sodium deoxycholate, but not by EDTA. The data indicate that different mechanisms may be involved in each of these coaggregations.

Molecular basis of bacterial adhesion in the oral cavity.

The two varieties of fimbriae identified on oral strains of actinomyces have distinct functional properties. The type 1 fimbriae of Actinomyces viscosus T14V mediate attachment to saliva-treated hydroxyapatite. Type 2 fimbriae--on A. viscosus and the only fimbriae detected on A. naeslundii WVU45--are associated with lectin activity. Interaction of these fimbriae with complementary receptors initiates bacterial attachment to Streptococcus sanguis 34 and sialidase-treated epithelial cells and the killing of actinomyces by polymorphonuclear leukocytes (PMNs). Galactose, N-acetylgalactosamine (GalNAc), and related oligosaccharides inhibit these processes, and mutants lacking type 2 fimbriae do not participate in them. The actinomyces lectin is similar to lectins from Ricinus communis and Bauhinia purpurea that agglutinate certain strains of oral streptococci, block attachment of actinomyces to epithelial cells, and inhibit killing of actinomyces by PMNs. The S. sanguis receptor for the actinomyces lectin comprises repeating hexasaccharide units with GalNAc termini. Used as probes, the peanut agglutinin, with specificity for Gal(beta-3)GalNAc, and the lectin from B. purpurea detect a 160-kilodalton (kdal) band in SDS-PAGE-separated epithelial cell extracts and a 100-kdal band in PMN extracts. These may be receptors for type 2 fimbriae. A. viscosus genes encoding subunits of types 1 and 2 fimbriae have been cloned in Escherichia coli; the type 1 subunit is 65 kdal and the type 2 subunit is 59 kdal. Submandibular immunization of mice with a mixture of type 1 and type 2 fimbriae evokes the production of IgA and IgG antibodies in serum and saliva that inhibit in vitro adsorption of A. viscosus to SHA. These antibodies may modulate colonization of teeth by this organism.

Actinomyces Adsorption Mediated by Type-1 Fimbriae

Monospecific antibody against the type-1 fimbriae of Actinomyces viscosus T14V reacted with 12 strains of A. viscosus (serotype 2), 12 strains of A. naeslundii (serotype 2 or serotype 3), but not with 11 serotype 1 strains of A. naeslundii. All strains positive for the type-1 fimbrial antigen adsorbed strongly to saliva-treated hydroxyapatite, and in many instances (18 of 24 strains) this interaction was inhibited by the monospecific antibody. These findings strongly suggest that only those strains of actinomyces which possess type-1 fimbriae adhere to the tooth surfaces, and that these fimbriae constitute the principal adhesin involved in this interaction.

Coagglutination reactions between Candida albicans and oral bacteria.

An agglutination assay for detecting intermicrobial adherence between the cells of Candida albicans and various oral bacteria is described. Strains of Streptococcus sanguis, S. salivarius, S. mutans, S. mitis, Fusobacterium nucleatum and Actinomyces viscosus all coagglutinated with C. albicans. No interaction could be demonstrated between the cells of Bacteroides melaninogenicus and those of C. albicans. Preliminary investigations of these interactions suggest that binding of F. nucleatum and A. viscosus to C. albicans is mediated by bacterial proteins, possibly lectins. Other mechanisms must account for the binding of oral streptococci to C. albicans. The possible implications of these findings in relation to oral mucosal colonisation and oral candidal clearance are discussed.

Root surface caries: a recurrent dilemma.

AbstractRoot surface caries, as a separate entity, has been recognized for a long time. It appears to be related to an altered bacteriology where some actinomyces strains allow decalcification of the cementum and dentine at a higher pH than the Streptococcus mutans usually associated with active caries. The problem is generally associated with the ageing patient and the causes are outlined. Early recognition and preventive measures such as the application of topical fluoride or chlorhexidine as an oral lavage are stressed. Appropriate dental treatment is discussed including restoration using glass‐ionomer cements.

Actinomyces slackii sp. nov. from Dental Plaque of Dairy Cattle

Five strains of actinomyces isolated from dental plaque of cattle formed a homogeneous group on the basis of Gram staining, colonial and cellular morphology, fermentation reactions, acid end products of metabolism, sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns of polypeptides, cell wall carbohydrates, and peptidoglycan and deoxyribonucleic acid mean base compositions. These organisms were closely related, as determined by deoxyribonucleic acid-deoxyribonucleic acid homology, which distinguishes them from strains of Actinomyces naeslundii, Actinomyces viscosus, Actinomyces denticolens, and Actinomyces howellii. A new taxon, Actinomyces slackii, is proposed for these isolates. The type strains of A. slackii is strain NCTC 11923.

Incidence and characterization of anti-microbial effects produced by Actinomyces viscosus and Actinomyces naeslundii.

Sixty-two facultative Actinomyces strains isolated from dental plaque were tested for the production of bacteriocin-like inhibitory effects by a deferred antagonism method. When incubated anaerobically, all isolates produced identical inhibitory patterns against 15 indicator organisms, but under microaerophilic conditions, little inhibitory activity was observed. Activity was not evident after anaerobic incubation on a medium buffered by 0.5% (w/v) calcium carbonate. Gas-liquid chromatographic analyses of agar blocks removed from the inhibitory zones indicated that, compared with microaerophilic conditions, anaerobic incubation encouraged production of high concentrations of lactic and succinic acids, and the concomitant fall in the pH was probably responsible for the inhibitory effects.