Competition for dietary carbohydrates between streptococci in dental plaque

Abstract The predominant microorganisms in dental plaque, i.e., streptococci and actinomycetes, are carbohydrate fermenters. In the natural environment these organisms experience carbohydrate limitation interrupted by periods of substrate excess following food intake by the host. Recently, we have studied the competition between pairs of oral Streptococcus species in the chemostat under continuous glucose or sucrose limitation and under glucose pulsed conditions.
In the present study we have investigated the competition for dietary carbohydrates in dental plaque of gnotobiotic rats infected with the same combinations of streptococci. The rats were exposed to similar regimes of carbohydrate administration as in the chemostat by incorporation of low molecular carbohydrates in the diet and drinking water. Addition of glucose to the diet favoured the organism with the highest q _max for glucose. This result parallels the outcome of earlier competition experiments in glucose-pulsed chemostats, which showed that the organism with the highest q _max glucose became dominant in the cultures. Previous observations that bacteriocin production and extracellular glucan synthesis are major ecological factors for oral streptococci were also confirmed in this experiment.     

Distribution of Streptococcus troglodytae and Streptococcus dentirousetti in chimpanzee oral cavities

The aim of this study was to analyze the distribution and phenotypic properties of the indigenous streptococci in chimpanzee (Pan troglodytes) oral cavities. Eleven chimpanzees (aged from 9 to 44 years, mean ± SD, 26.9 ± 12.6 years) in the Primate Research Institute of Kyoto University were enrolled in this research and brushing bacterial samples collected from them. Streptococci were isolated from the oral cavities of all chimpanzees. The isolates (n = 46) were identified as thirteen species by 16S rRNA genes analysis. The predominant species was Streptococcus sanguinis of mitis streptococci from five chimpanzees (45%). Mutans streptococci were isolated from six chimpanzees (55%). The predominant species in the mutans streptococci were Streptococcus troglodytae from four chimpanzees (36%), this species having been proposed as a novel species by us, and Streptococcus dentirousetti from three chimpanzees (27%). Streptococcus mutans was isolated from one chimpanzee (9%). However, Streptococcus sobrinus, Streptococcus macacae and Streptococcus downei, which are indigenous to human and monkey (Macaca fasciclaris) oral habitats, were not isolated. Of the mutans streptococci, S. troglodytae, S. dentirousetti, and S. mutans possessed strong adherence activity to glass surface.     

Competition between different strains of Streptococcus cremoris

Abstract Streptococcus cremoris strain HP was found to grow poorly on agar plates under aerobic conditions in comparison to several other strains of S. cremoris (Wg2, ML1, AM1, E8). This made it possible to determine the numbers of strain HP in mixed cultures with other strains under different culture conditions. None of the mixtures was stable in batch cultures as a result of differences in the maximum specific growth rate. In continuous culture under lactose limitation strain HP outcompeted strains E8 and ML1 at low dilution rates, but at high dilution rates and in batch culture the reverse was observed. This represents another example of crossing μ-s curves in anaerobic bacteria.     

Regulation of glucose catabolism in Thiobacillus A2 grown in the chemostat under dual limitation by succinate and glucose

In contrast to its diauxic behaviour in batch culture, Thiobacillus A2 grew in chemostat culture using glucose and succinate as dual limiting substrates. Biomass production under dual limitations was the sum of that on single substrates with each substrate being oxidized and assimilated to similar extents in single and dual substrate-limited cultures. In glucose and glucose + succinate-limited cultures glucose was oxidized largely by the Entner-Doudoroff and pentose phosphate pathways, but other mechanisms also contributed and the ratios of pathways depended on substrate ratios and the previous substrate-history of the culture. Variations in specific activities of enzymes of carbohydrate metabolism following switches from single to mixed substrates were considerable, ranging from fourfold for fructose diphosphate aldolase to more than 200-fold for hexokinase, fructose diphosphatase, glucose 6-phosphate and 6-phosphogluconate dehydrogenases. Changes in specific activities occurred only over prolonged time periods in the chemostat, probably reflecting low concentrations of free substrates in carbon-limited cultures and consequent low levels of catabolite repression.     

Effect of saliva viscosity on the co-aggregation between oral streptococci and Actinomyces naeslundii

doi: 10.1111/j.1741‐2358.2011.00595.x Effect of saliva viscosity on the co‐aggregation between oral streptococci and Actinomyces naeslundii Background: The co‐aggregation of oral bacteria leads to their clearance from the oral cavity. Poor oral hygiene and high saliva viscosity are common amongst the elderly; thus, they frequently suffer from pneumonia caused by the aspiration of oral microorganisms. Objectives: To examine the direct effect of saliva viscosity on the co‐aggregation of oral streptococci with actinomyces. Materials and methods: Fifteen oral streptococcal and a single actinomyces strain were used. Co‐aggregation was assessed by a visual assay in phosphate buffer and a spectrophotometric assay in the same buffer containing 0–60% glycerol or whole saliva. Results: Nine oral streptococci co‐aggregated with Actinomyces naeslundii ATCC12104 in the visual assay and were subsequently used for the spectrophotometric analysis. All tested strains displayed a decrease in co‐aggregation with increasing amounts of glycerol in the buffer. The co‐aggregation of Streptococcus oralis with A. naeslundii recovered to baseline level following the removal of glycerol. The per cent co‐aggregation of S. oralis with A. naeslundii was significantly correlated with the viscosity in unstimulated and stimulated whole saliva samples (correlation coefficients: ?0.52 and ?0.48, respectively). Conclusion: This study suggests that saliva viscosity affects the co‐aggregation of oral streptococci with actinomyces and that bacterial co‐aggregation decreases with increasing saliva viscosity.     

Competition for L-glutamate between specialised and versatile Clostridium species

Clostridium cochlearium could be reproducibly enriched in an L-aspartate- and L-glutamate-limited, anaerobic chemostat inoculated with anaerobic sludge. L-glutamate, L-glutamine and L-histidine were the only fermentable substrates. Less specialised clostridia of the C. tetanomorphum type could only be isolated from batch enrichments with L-glutamate and L-aspartate as energy sources. Competition experiments with C. cochlearium and C. tetanomorphum in a L-glutamate-limited chemostat resulted in the selective elimination of the latter species. Addition of glucose to the medium resulted in coexistence of both species. The molar growth yields for L-glutamate at different dilution rates at 30°C were determined for both species. The maximum specific growth rates on L-glutamate were 0.55 h^-1 for C. cochlearium and 0.35 h^-1 for C. tetanomorphum.     

Long run coexistence in the chemostat with multiple species

In this work we analyze the transient behavior of the dynamics of multiple species competing in a chemostat for a single resource, presenting slow/fast characteristics. We prove that coexistence among a subset of species, with growth functions close to each other, can last for a substantially long time. For these cases, we also show that the proportion of non-dominant species can be increasing before decreasing, under certain conditions on the initial distribution.     

A lacticin 3147 enriched food ingredient reduces Streptococcus mutans isolated from the human oral cavity in saliva

AIMS: To isolate and characterise Streptococcus mutans from Irish saliva samples and to assess their sensitivity to a food-grade preparation of the lantibiotic, lacticin 3147, produced by Lactococcus lactis DPC3147. METHODS AND RESULTS: Saliva samples collected from children with varying oral health status were screened on Mitis Salivarius agar for the presence of pathogenic streptococci. Following selective plating, 16S rDNA sequencing and Pulsed Field Gel Electrophoresis (PFGE), 15 distinct strains of Strep. mutans were identified. These were grouped according to their relative sensitivity to lacticin 3147 which ranged from 0.78 to 6.25%; relative to a sensitive indicator strain, Lactococcus lactis ssp. lactis HP. Inhibition of indicator Strep. mutans strains from sensitive, intermediate and tolerant groupings were assessed in microtitre plate assays with increasing concentrations of lacticin 3147. The concentration of lacticin 3147 required to give 50% growth inhibition correlated with their relative sensitivities (as assayed by well diffusion methodology) and ranged from 1280 to 5120 AU ml(-1). Concentrated preparations of lacticin 3147 caused a rapid killing of Strep. mutans strains in broth. Moreover, in human saliva deliberately spiked with Strep. mutans, the pathogen was eliminated (initial inoculum of 10(5)) in the presence of 40,000 AU ml(-1) of lacticin 3147. Furthermore, a food-grade lacticin 3147 spray dried powder ingredient was assessed for the inhibition of Strep. mutans in human saliva, spiked with a strain of intermediate sensitivity, resulting in up to a 4-log reduction in counts after 20 min. CONCLUSION: A food grade preparation of lacticin 3147 was effective in the inhibition of oral Strep. mutans. SIGNIFICANCE AND IMPACT OF THE STUDY: The inhibition of oral streptococci by food grade preparations of lacticin 3147 may offer novel opportunities for the development of lacticin 3147 as an anti-cariogenic agent particularly in the area of functional foods for the improvement of oral health.     

Growth of Saccharomyces cerevisiae in a chemostat under high glucose conditions

Saccharomyces cerevisiae was grown in a chemostat under high glucose conditions (up to 300 g l^–1). The results support the view that higher glucose feed favors higher ethanol production regardless of the existence of osmotic stress. A low glucose utilization and yield coefficient provides an opportunity to improve continuous fermentation performance in the fuel alcohol industry. The possibility exists of reusing yeast cells and subsequently lower operating costs, and by using an optimal glucose feeding concentration between 100 and 200 g l^–1.     

Human oral epithelial cell culture I. Improved conditions for reproducible culture in serum-free medium

Summary Gingival tissue from healthy adult human donors was used as a source of epithelial cells for culture. An overnight incubation of this tissue with dispase facilitated the mechanical separation of the surface epithelium from the underlying fibrous connective tissue. This step minimized culture contamination with fibroblasts. The epithelium was then trypsinized to prepare a single cell suspension. The cell pellets were collected by centrifugation and resuspended in keratinocyte growth medium, incubated at 37° C and 5% CO₂ in a humid atmosphere. Primary cultures grew in small islands that coalesced at confluency. Immunohistochemistry demonstrated uniform staining of the cells with antibodies to keratins of stratified squamous epithelium. Ultrastructurally, the cells contained distinct intermediate filaments. When cells were grown in media with low calcium (0.15 mM), cell-to-cell contacts were via interlacing papillary projections with no desmosomes. However, when cells were grown under physiologic calcium (1.2 mM), desmosomes were prominent and well developed. Cells were maintained in culture for over 100 d (7 passages). This work was supported by Biomedical Research grant RR 05346 from the National Institute of Health, Bethesda, MD, and the University of Washington Graduate School Research Fund.     

Toxin production and competitive abilities under phosphorus limitation of Alexandrium species

The relationship between growth rate versus phosphorus concentration and cellular toxin content was determined for Alexandrium minutum AL1V, Alexandrium tamarense MDQ1096, A. tamarense EF04 and Alexandrium andersoni EF12 under different nitrogen and phosphorus supplies. The aim was to determine whether those species with a lower phosphorus uptake affinity, and hence potentially of lower competitive ability at low phosphorus concentrations, were more toxic. The range and mean of toxic content per cell (as fmol per cell) of the species were 13.5–256.5 and 140.2±50.8 for A. tamarense MDQ1096, 0.5–16.5 and 2.9±2.6 for A. minutum, 0–2.0 and 0.2±0.3 for A. tamarense EF04 and, 0–3.3 and 0.06±0.4 for A. andersoni. K_s for culture cell growth (per day),representing the phosphate concentration at which the specific culture cell growth rate is one half the maximum rate, and K_min (per day), the phosphate concentration at which the specific culture cell growth rate is zero, were used as indicators of species’ potential competitive ability at low phosphorus concentrations. Low values for both K_s and K_min indicate a high relative ability of the species to use low levels of phosphate and, hence, expected to outcompete higher K_s and K_min species under phosphorus limitation. K_s and K_min were 1.68 and 0.48 for A. tamarense MDQ1096, 1.16 and 0.39 for A. minutum, 1.0 and 0.38 for A. tamarense EF04 and, 0.74 and 0.34 for A. andersoni, respectively. There was a significant positive relationship between toxin content per cell with both K_s and K_min, indicating that those species with lower ability to compete under phosphorus limitation were more toxic. The findings support the hypothesis that toxin production by dinoflagellates species could be an adaptation evolved to offset the ecological disadvantage of having low nutrient affinity.     

Mixed chemostat cultures of obligately aerobic and fermentative or methanogenic bacteria grown under oxygen-limiting conditions

Abstract Defined mixed cultures of an obligately aerobic Pseudomonas testosteroni and anaerobic Veillonella alcalescens strain were grown under oxygen and lactate limitation in chemostats with different oxygen supply rates. The aerobic and the anaerobic bacteria were shown to coexist and to complete for common substrates over a wide range of oxygen supply rates. Under similar conditions but with formate as the major substrate chemostat enrichments gave rise to undefined mixed cultures of aerobic, fermentative and methanogenic bacteria. The relevance of these observations to natural mineralization processes is discussed.     

Relationship between the ability of oral streptococci to interact with platelet glycoprotein Ibalpha and with the salivary low-molecular-weight mucin, MG2

The oral streptococci Streptococcus sanguinis, Streptococcus gordonii and Streptococcus oralis are common aetiological agents of infective endocarditis, and their ability to adhere to and induce the aggregation of platelets is thought to be a virulence trait. The platelet glycoprotein GPIbalpha has been implicated as the adhesion receptor for S. sanguinis and S. gordonii, but it is not known if this is the case for S. oralis and other species. The aim of this study was to determine the GPIbalpha-interactive capability of a range of oral streptococci and to determine the relationship between this capability and their ability to interact with the salivary constituents that they would encounter in their normal habitat. All platelet-adhesive S. sanguinis strains and most S. gordonii strains adhered in a GPIbalpha-dependent manner, but strains of S. oralis, Streptococcus cristatus, Streptococcus parasanguinis and Streptococcus mitis had no direct affinity for platelets. Those strains that were able to bind GPIbalpha also bound to the low-molecular-weight submandibular salivary mucin, MG2, and this interaction was sialic acid-dependent. The data suggest that S. sanguinis and S. gordonii may be efficient colonizers of platelet vegetations because of their adaptation to recognize sialylated salivary mucins. In contrast, S. oralis does not interact with platelets and so is likely to colonize vegetations through an as yet unidentified mechanism.     

Practical coexistence of two species in the chemostat - A slow-fast characterization

We show that the chemostat model with two species having different but close break-even concentrations exhibits a slow-fast dynamics. Considering small perturbations about the dilution rate for which break-even concentrations are identical, we use the Fenichel theory to show the coexistence of species for large times. Then we determine the reduced dynamics, which is non-trivial and characterized by the slopes of the growth functions about their break-even concentrations.     

Studies on interspecies competition between the azuki bean weevil,Callosobruchus chinensis,and the southern cowpea weevil,C. maculatus II. Competition under different environmental conditions

The experimental studies was done on the interspecies competition between the azuki bean weevil, Callosobruchus chinensis, and the southern cowpea weevil, C. maculatus, under the different environmental conditions.

1. At 30°C, 70% R. H., the change of food replenishment interval had no influence upon the competition results, and the competition always ended in the extinction of C. maculatus. But there was a tendency for the period of co-existence to be elongated as the interval of replenishment lengthened.
2. At 30°C, 70% R. H., there were no differences in the competition processes and results between the competition using the dry bean (water content about 11%) or the normal one (about 15%).
3. At 32°C, 64% R. H., the competition result was reversed from that at 30°C, 70% R. H. This is thought to be due to the differences of the developmental rates and the numbers of eggs oviposited between the two species at these conditions.
4. In some cases the competition result could be understood from the point of the phenomena of single species populations of both species, but in other cases, the results were in conflict with the expectation from those phenomena. To analyze the competition mechanism, it is necessary to investigate the interaction between species.

     

Resource limitation,habitat segregation,and species interactions of british tree-hole mosquitoes in nature

Summary The insect fauna of water-filled tree holes in southern Britain consists primarily of the mosquitoes Aedes geniculatus, Anopheles plumbeus, Culex torrentium, and a benthic detritivorous fauna that includes primarily the scirtid beetle Prionocyphon serricornis and the chironomid midge Metriocnemus martinii. Culex torrentium has been documented only relatively recently in tree holes but all three species of mosquitoes partition the resource in space and time. When mosquito larvae were forced to coexist in natural tree holes at limiting densities and at higher than natural levels of interspecific encounter, there was no evidence that Aedes geniculatus or Anopheles plumbeus affected pupation success, pupal weight, or development time of the other or that either Aedes geniculatus or C. torrentium affected the survivorship, pupation success, pupal weight, and biomass yield of the other. When A. geniculatus at limiting densities were forced in natural tree holes to live without or to coexist with natural or twice natural densities of P. serricornis and M. martinii, the presence, absence, or superabundance of the benthic insects did not affect pupation success or pupal weight of A. geniculatus; development time of A. geniculatus was faster when a superabundance of the benthic fauna was present. Effects of the benthic fauna on A. geniculatus are slight and the only significant interaction is facilitative, not competitive. The pattern of habitat segregation among treehole mosquitoes in southern Britain is characteristic of their respective genera and we propose that this pattern is more likely (but not certain) to have arisen through a process of independent evolution than through competitively driven niche shifts among already coexisting species.     

Competition between sulfate-reducing and methanogenic bacteria for H2 under resting and growing conditions

The basis for the outcome of competition between sulfidogens and methanogens for H₂ was examined by comparing the kinetic parameters of representatives of each group separately and in co-culture. Michaelis-Menten parameters (V _max and K _m) for four methanogens and five sulfate-reducing bacteria were determined from H₂-depletion data. Further, Monod growth parameters (_max, K _s, Y _H2) for Desulfovibrio sp. G11 and Methanospirillum hungatei JF-1 were similarly estimated. H₂ K _m values for the methanogenic bacteria ranged from 2.5 M (Methanospirillum PM1) to 13 M for Methanosarcina barkeri MS; Methanospirillum hungatei JF-1 and Methanobacterium PM2 had intermediate H₂ K _m estimates of 5 M. Average H₂ K _m estimates for the five sulfidogens was 1.2 M. No consistent difference among the V _max estimates for the above sulfidogens (mean=100 nmol H₂ min^-1 mg^-1 protein) and methanogens (mean=110 nmol H₂ min^-1 mg^-1 protein) was found. A two-term Michaelis-Menten equation accurately predicted the apparent H₂ K _m values and the fate of H₂ by resting co-cultures of sulfate-reducers and methanogens. Half-saturation coefficients (K _s) for H₂-limited growth of Desulfovibrio sp. G11 (2–4 M) and Methanospirillum JF-1 (6–7 M) were comparable to H₂ K _m estimates obtained for these organisms. Maximum specific growth rates for Desulfovibrio sp. G11 (0.05 h^-1) were similar to those of Methanospirillum JF-1 (0.05–0.06 h^-1); whereas G11 had an average yield coefficient 4 x that of JF-1. Calculated _max and V _max/K _m values for the methanogens and sulfidogens studied predict that the latter bacterial group will process more H₂ whether these organisms are in a growing or resting state, when the H₂ concentration is in the first-order region.