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D C Ellwood

Publications and source records attributed to D C Ellwood.

At least 19 recordsLinked to original sources

Comparison of properties of collected cells and cells from the culture vessel during continuous culture of Streptococcus mutans Ingbritt.

In continuous-culture studies chemostat effluents are usually collected into a receiving flask in an ice bath to obtain enough cells for an experiment. It is assumed that the properties of these are not significantly different from those of the culture in the chemostat vessel. This assumption has been tested for the dental pathogen Streptococcus mutans Ingbritt. Collected supernatant fluid and cells were compared with supernatant fluid and cells taken directly from the culture vessel, for four major groups of culture properties: viability and biomass, concentrations of metabolites and nutrients, activities of selected enzymes, and glycolytic rates. The assumption held true except for glycolytic rate during endogenous metabolism. It is suggested that comparison of collected and culture vessel cells is an important control which should be done in all continuous culture studies of microbial physiology and biochemistry, but that the properties of Strep. mutans cells collected on ice up to 16 h do reflect those of cells actively growing in the chemostat.

Bacteriological Techniques↗

Protonmotive force driven 6-deoxyglucose uptake by the oral pathogen, Streptococcus mutans Ingbritt.

Streptococcus mutans Ingbritt was grown in glucose-excess continuous culture to repress the glucose phosphoenolpyruvate phosphotransferase system (PTS) and allow investigation of the alternative glucose process using the non-PTS substrate, (3H) 6-deoxyglucose. After correcting for non-specific adsorption to inactivated cells, the radiolabelled glucose analogue was found to be concentrated approximately 4.3-fold intracellularly by bacteria incubated in 100 mM Tris-citrate buffer, pH 7.0. Mercaptoethanol or KCl enhanced 6-deoxyglucose uptake, enabling it to be concentrated internally by at least 8-fold, but NaCl was inhibitory to its transport. Initial uptake was antagonised by glucose but not 2-deoxyglucose. Evidence that 6-deoxyglucose transport was driven by protonmotive force (delta p) was obtained by inhibiting its uptake with the protonophores, 2,4-dinitrophenol, carbonylcyanide m-chlorophenylhydrazine, gramicidin and nigericin, and the electrical potential difference (delta psi) dissipator, KSCN. The membrane ATPase inhibitor, N,N1-dicyclohexyl carbodiimide, also reduced 6-deoxyglucose uptake as did 100 mM lactate. In combination, these two inhibitors completely abolished 6-deoxyglucose transport. This suggests that the driving force for 6-deoxyglucose uptake is electrogenic, involving both the transmembrane pH gradient (delta pH) and delta psi. ATP hydrolysis, catalysed by the ATPase, and lactate excretion might be important contributors to delta pH.

Deoxy Sugars↗

The effect of lowering the pH on the composition and metabolism of a community of nine oral bacteria grown in a chemostat.

Nine oral bacteria, associated with both healthy and diseased sites in the mouth, were grown at D = 0.05 h-1 (mean generation time 13.9 h) in a glucose-limited chemostat. After an initial period of steady-state growth at pH 7.0, pH control was discontinued. The pH then decreased until it stabilized at pH 4.1 after 9 d (16 generations), while the Eh rose from -165 mV to +160 mV. The lowering in pH resulted in the composition and metabolism of the flora being altered and in increased bacterial aggregation. At pH 7.0, 'Streptococcus mitior', Veillonella alcalescens and S. sanguis were most numerous while at pH 4.1 the counts of all bacteria fell except for Lactobacillus casei, which became predominant. The proportions of S. mutans within the community also increased while S. sanguis was recovered only occasionally and Bacteroides intermedius was not detected below pH 4.6. The survival at pH 4.1 of several other species would not have been predicted from earlier pure culture studies. Relative to pH 7.0, the community growing at pH 4.1 produced more lactic acid, washed cells had a greater glycolytic activity over a wider pH range but amino acid metabolism decreased. In general, when pH control was restored, so were the original patterns of metabolism and bacterial counts, except for B. intermedius, which was still not detected. The inverse relationship between S. sanguis and S. mutans, and the increase in proportions of L. casei and S. mutans during growth in a low pH environment parallel observations made in vivo and suggest that the chemostat can be used as a model for microbial behaviour in dental plaque.

Actinomyces↗

Effect of hemin on the physiology and virulence of Bacteroides gingivalis W50.

Bacteroides gingivalis W50 was grown in a chemostat under steady-state conditions at pH 7.5 +/- 0.2 and a constant growth rate of 6.9 h for periods of up to 6 weeks (146 bacterial generations) in a complex medium. Hemin was capable of limiting the growth of cells up to a concentration of approximately 0.5 micrograms/ml since higher concentrations of hemin did not increase cell yields; cells grew in the absence of exogenously added vitamin K1. Only a limited number of amino acids was metabolized during growth, but because none of these was totally depleted, the limiting nutrient under hemin excess conditions was probably a peptide. A range of fermentation products was produced under all conditions of growth; higher concentrations of cytotoxic metabolites such as propionate and butyrate were formed under hemin excess conditions, although more ammonia was released under hemin limitation. When viewed by electron microscopy, cells grown under hemin limitation appeared to be either coccobacillary or short rods and possessed few fimbriae per cell, but large numbers of extracellular vesicles could be seen both surrounding the cell surface and free in the environment. In contrast, cells grown under hemin excess conditions were more commonly coccus shaped and were more heavily fimbriated but had fewer extracellular vesicles. Marked differences were found in the susceptibility of mice to infection with cells grown under different concentrations of hemin. Cells transferred to media without any added hemin were avirulent, whereas those grown under conditions of hemin limitation (0.33 and 0.40 micrograms/ml) produced a 20 and 50% mortality in mice, respectively. In contrast cells grown under hemin excess always caused 100% mortality in mice, although this virulence was dose dependent. When virulent, the bacteria caused an extensive, spreading infection with necrosis of the skin and subcutaneous tissues. Collagen disintegration was seen histologically, implying a role for collagenase production in the pathogenicity of these bacteria.

Animals↗

Environmental regulation of carbohydrate metabolism by Streptococcus sanguis NCTC 7865 grown in a chemostat.

Carbohydrate metabolism by the oral bacterium Streptococcus sanguis NCTC 7865 was studied using cells grown in a chemostat at pH 7.0 under glucose or amino acid limitation (glucose excess) over a range of growth rates (D = 0.05 h-1-0.4 h-1). A mixed pattern of fermentation products was always produced although higher concentrations of lactate were formed under amino acid limitation. Analysis of culture filtrates showed that arginine was depleted from the medium under all conditions of growth; a further supplement of 10 mM-arginine was also consumed but did not affect cell yields, suggesting that it was not limiting growth. Except at the slowest growth rate (D = 0.05 h-1) under glucose limitation, the activity of the glucose phosphotransferase (PTS) system was insufficient to account for the glucose consumed during growth, emphasizing the importance of an alternative method of hexose transport in the metabolism of oral streptococci. The PTS for a number of sugars was constitutive in S. sanguis NCTC 7865 and, even though the cells were grown in the presence of glucose, the activity of the sucrose-PTS was highest. The glycolytic activity of cells harvested from the chemostat was affected by the substrate, the pH of the environment, and their original conditions of growth. Glucose-limited cells produced more acid than those grown under conditions of glucose excess; at slow growth rates, in particular, greater activities were obtained with sucrose compared with glucose or fructose. Maximum rates of glycolytic activity were obtained at pH 8.0 (except for cells grown at D = 0.4 h-1 where values were highest at pH 7.0), while slow-growing, amino acid-limited cells could not metabolize at pH 5.0. These results are discussed in terms of their possible significance in the ecology of dental plaque and the possible involvement of these bacteria in the initiation but not the clinical progression of a carious lesion.

Amino Acids↗

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

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

Actinomyces↗

Effect of environmental conditions on the fluoride sensitivity of acid production by S. sanguis NCTC 7865.

Growth and environmental conditions affected the fluoride (F) sensitivity of acid production by Streptococcus sanguis NCTC 7865. Cells grown glucose-limited in a chemostat were generally more sensitive than those harvested from cultures in which there was an excess of glucose (amino acid-limited). There was no consistent relationship between the growth rate of cells and their F sensitivity. Slower-growing cells (mean generation time = 14 hr) were more sensitive than those growing quickly when glucose was the limiting nutrient, whereas the faster growing cells from the glucose-excess culture were most susceptible. The pH of the environment markedly affected the F sensitivity of cells: 2 mM F- was sufficient to abolish acid production by cells incubated at pH 5.0, whereas 24 mM F- did not totally inhibit glycolysis at pH 7.0 or 8.0. Regardless of pH and growth conditions, the cationic composition of the environment had the most pronounced effect on acid production and fluoride sensitivity. Cells washed and re-suspended in KCl were more acidogenic and more sensitive to F than the same cells treated with saline. At pH 7.0 and 8.0, saline-washed cells were comparatively unaffected by F, while glycolysis by the same cells at the same pH but washed in KCl could be inhibited by up to 80%. These results suggested that F inhibition could not be explained merely on the basis of HF uptake at low pH values. Since it has been shown previously that the activity of the energized membrane is maintained by K+ and dissipated in the presence of Na+, it was proposed that proton motive force (pmf) might be involved in the uptake of F-.

Acids↗

Evidence that glucose and sucrose uptake in oral streptococcal bacteria involves independent phosphotransferase and proton-motive force-mediated mechanisms.

Sugar transport and glycolysis in Streptococcus sanguis NCTC 7865, Streptococcus mitis ATCC 903, Streptococcus salivarius NCTC 8606 and several strains of Streptococcus mutans were investigated by following the rate of acid production by washed bacteria at a constant pH of 7.0. The phosphoenolpyruvate-phosphotransferase system (PTS) was inhibited by low concentrations of chlorhexidine. When this PTS-inhibitory concentration of chlorhexidine was added to cells washed and re-suspended in KCl, glucose uptake and glycolysis continued at a greatly-reduced rate. Chlorhexidine abolished glucose and sucrose uptake and metabolism in bacteria washed and incubated in saline. The Na+-inhibition was reproduced in KCl-washed bacteria using the cyclic peptide ionophores, valinomycin and gramicidin, to dissipate K+ and H+ gradients across the cell membrane. Glucose metabolism by Strep. mutans B13 was more resistant to chlorhexidine than that of Strep. mutans NCTC 10449 or Strep. sanguis but was more sensitive to the ionophores. Valinomycin had a greater inhibitory effect on strain B13 than the other two. That ion gradients are important in the chlorhexidine-resistant glucose-uptake mechanism was confirmed using the classical uncoupling agents, carbonylcyanide-m-chlorophenylhydrazone, 2,4-dinitrophenol and KSCN. Glucose metabolism was inhibited in the presence of both the uncouplers and the PTS-inhibitory concentration of chlorhexidine and significant inhibition was also observed in the absence of the PTS inhibitor. Lactate or the ATPase inhibitor, dicyclohexyl carbodiimide (DCCD), had similar inhibitory effects on the non-PTS uptake system.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphatases↗

Regulation of glucose metabolism in oral streptococci through independent pathways of glucose 6-phosphate and glucose 1-phosphate formation.

In vivo rates of glucose uptake and acid production by oral streptococci grown in glucose- or nitrogen-limited continuous culture and batch culture were compared with the glucose phosphorylation activities of harvested, decryptified cells. The strains examined contained significant phosphoenolpyruvate-phosphotransferase system (PTS) activity, measured by a glucose 6-phosphate (G6P) dehydrogenase-linked assay procedure, but this activity was insufficient to account for the in vivo glucose uptake rates. However, ATP was a superior phosphoryl donor to phosphoenolpyruvate, and unlike the PTS, phosphoryl transfer with ATP was insensitive to bacteriostatic concentrations of chlorhexidine, suggesting glucokinase-mediated G6P formation. Again, G6P formation from the PTS and glucokinase reactions was not commensurate with some of the glucose uptake rates observed, implying that other phosphorylation reactions must be occurring. Two novel reactions involving carbamyl phosphate and acetyl phosphate were identified in some of the strains. No G6P formation was detected with these potential phosphoryl donors, but in the presence of phosphoglucomutase, glucose 1-phosphate (G1P) formation was evident, which was insensitive to chlorhexidine. G1P is a precursor of glycogen, and good correlation was obtained between G1P formation activity and endogenous metabolism of washed cells measured either as a rate of acid production at a constant pH 7 or as a decrease in pH with time in the absence of titrant. A "league table" of abilities to synthesize G1P and produce acid from endogenous metabolism was compiled for oral streptococci grown in batch culture. This indicated that Streptococcus mutans Ingbritt and Streptococcus sanguis Challis were unable to form G1P or produce much acid endogenously, whereas increasing activities were obtained with Streptococcus salivarius, Streptococcus sanguis, and Streptococcus mitis. In particular, S. mitis had the highest G1P formation activities and was able to decrease the pH to less than 5 in 15 min by endogenous metabolism alone. The data are consistent with the intracellular accumulation of free glucose driven by proton motive force when PTS activities are low and the subsequent phosphorylation to either G6P for metabolism via glycolysis or G1P for glycogen biosynthesis. The accumulation of acetyl phosphate during glucose-limited growth and the availability of arginine for catabolism to carbamyl phosphate provide an explanation as to why some glucose-limited oral streptococci continue to synthesize glycogen under these conditions, which might prevail in plaque.

Chlorhexidine↗

Relationship of bioenergetic processes to the pathogenic properties of oral bacteria.

The energized membrane has been shown to affect properties (sugar transport, acid production, intracellular polysaccharide formation, and glycosyltransferase secretion) related to the pathogenicity of oral bacteria. The activity of the energized membrane was susceptible to modulation by environmental conditions likely to be encountered by bacteria in dental plaque.

Acids↗

Inhibition by the antimicrobial agent chlorhexidine of acid production and sugar transport in oral streptococcal bacteria.

Oral streptococci transport sugars via the phosphoenolpyruvate-phosphotransferase (PEP-PTS) system. In a specific assay of this system, low concentrations of chlorhexidine abolished the activity of the glucose and sucrose PTS in batch-grown cells of Streptococcus mutans Ingbritt and B13, Strep. sanguis NCTC 7865, Strep. mitis ATCC 903, Strep. milleri NCTC 10709 and Strep. salivarius NCTC 8606. Intact cells and cells made permeable to the assay reagents with toluene were used. Toluenized cells were more sensitive to chlorhexidine than intact cells (0.09 and 0.25 mM, respectively). This PTS-inhibitory concentration of chlorhexidine reduced acid production from glucose in pH fall experiments to values higher than are obtained solely from endogenous metabolism. The effect of chlorhexidine on rates of acid production was determined at pH 7.0 using cells washed with either 135 mM NaCl or 135 mM KCl. In general, faster rates of acid production from the metabolism of glucose and sucrose were obtained with potassium-treated cells. Addition of the PTS-inhibitory concentration of chlorhexidine markedly reduced or totally abolished acid production by NaCl-treated cells; a greater residual-activity was detected in the same cells washed with KCl (except with Strep. mutans B13 and Strep. mitis ATCC 903). The PTS-inhibitory concentration of chlorhexidine had little or no effect on the viability of cells. The results confirm the existence of sugar uptake systems in oral streptococci additional to the PTS and provide an explanation for the additive anti-caries effect of mouth-rinses containing both fluoride and chlorhexidine.

Acids↗

The influence of growth rate and nutrient limitation on the microbial composition and biochemical properties of a mixed culture of oral bacteria grown in a chemostat.

A sample of human dental plaque was homogenized in transport fluid and inoculated simultaneously into a glucose-limited and a glucose-excess chemostat maintained at pH 7.0 and a dilution rate (D) of 0.05 h-1. In an attempt to ensure the establishment of slow-growing bacterial populations, two further inoculations of each chemostat with fresh samples of dental plaque took place before a steady-state was attained at this dilution rate. The dilution rate was increased step-wise to D = 0.6 h-1, and then returned directly to D = 0.05 h-1. Contrary to chemostat theory, microbial communities with a high species diversity were maintained under all of the experimental conditions employed, although not all of the bacterial populations present in the inocula established successfully in the chemostat. At each steady-state the bacteriological composition and biochemical properties (fermentation products, enzyme assays and acid production) of the communities of each chemostat was determined. Higher cell yields and a slightly more diverse community were obtained from the glucose-limited chemostat at all dilution rates. A complex mixture of end products of metabolism was obtained from the glucose-limited chemostat, suggesting amino acid catabolism, while lactate was the predominant acid of the glucose-excess culture. In washed-cell experiments, communities from the glucose-excess chemostat produced the lower terminal pH values following a pulse of glucose, with the lowest pH values occurring at the higher dilution rates. A film of micro-organisms, which accumulated around the neck of the chemostat, was sampled at the end of the experiment. The microbial composition of the films from each chemostat differed markedly, and both were different to the community of the bulk fluid of the respective chemostat. Spirochaetes and a population of yeasts were detected in the films from the glucose-limited and glucose-excess chemostats, respectively. No invertase or glucosyltransferase activity, and little glucoamylase-specific glycogen was detected in the communities from either chemostat, although significant endogenous activity, particularly at high dilution rates, was obtained with washed-cells from the glucose-excess chemostat. The results suggest that the chemostat could make a valuable contribution to the study of the ecology of dental plaque.

Bacteria↗

Carbohydrate metabolism by Actinomyces viscosus growing in continuous culture.

A human oral strain of Actinomyces viscosus, GN431/75, was grown anaerobically in a defined medium in continuous culture with a glucose limitation at dilution rates (D) between 0.025 and 0.2 h-1 and with a nitrogen limitation at D = 0.005 and 0.1 h-1. With 5 mg of glucose per ml, the culture was limited for carbon at D = 0.025 and 0.05 h-1, but became nitrogen limited (asparagine) at D = 0.1 and 0.2 h-1. The molar growth yield (Yglucose) decreased from 50.0 to 40.9 g of cells per mol of glucose as the dilution rate was increased from 0.025 to 0.2 h-1, reflecting the limitation of asparagine. With high glucose and low amino acid concentrations (nitrogen limited), the cell yields at D = 0.05 and 0.1 h-1 were 37 to 33% lower than in the glucose-limited culture. The major products of metabolism were succinic and lactic acids with lesser amounts of acetic and formic acids and ethanol. The rate of glucose fermentation by resting cells removed from the glucose-limited culture and assayed in a pH stat increased with the dilution rate and was always higher than that for the fermentation of sucrose (60%) and fructose (40%). Activity for the glucose-P-enolpyruvate phosphotransferase system was observed in whole homogenates, with the highest activity evident at D = 0.1 h-1 with the glucose-limited culture. The observed activity was significantly lower than the rate of glucose metabolism at each dilution rate, suggesting that glucose-P-enolpyruvate phosphotransferase system was underestimated or that an additional transport system exists in the organism. The glucose-limited culture showed considerable ability to synthesize glycogen during the transition from carbon to nitrogen limitation, when 35% of the cell mass was present at this polymer. The organism was shown to possess the glycogen synthetic enzymes ADP glucose synthase and ADP glucose transferase, as well as the degradative enzyme maltodextrin phosphorylase. Washed cells of A. viscosus GN431/75 were shown to be relatively insensitive to the inhibiting actions of NaF in pH-fall and constant-pH experiments at all dilution rates. At pH 7.0, 25 mM NaF was required to completely inhibit glycolysis by glucose-limited cells at D = 0.05 h-1, whereas a concentration of only 11 mM NaF was required with cells of Streptococcus mutans grown and incubated under identical conditions. An interesting feature of the growth of A. viscosus GN431/75 in the chemostat was the shift from individual nonadherent cells at the low dilution rates to the appearance at D = 0.2 h-1 of large cell aggregates forming tenacious adherent films reminiscent of its characteristics in the oral cavity.

Actinomyces↗

Surface-associated growth.

In natural ecosystems, microbial activity is often associated with the presence of a surface, particularly in low-nutrient environments. The chemostat allows the study of such low-nutrient environments together with the precise control of other growth parameters. By using this system, enrichment cultures with inocula from two different river sources have been made. A more diverse community attached itself to surfaces placed in the chemostat when the cultures were carbon-limited than when the limiting nutrient was nitrogen. Further studies on a pseudomonad isolated from the carbon-limited enrichment cultures have shown that surface-associated organisms grow at approximately twice the rate of the same organism in the free surrounding medium. A hypothesis to explain this phenomenon based on the chemiosmotic theory is discussed.

Bacteria↗

Influence of sodium and potassium ions on acid production by washed cells of Streptococcus mutans ingbritt and Streptococcus sanguis NCTC 7865 grown in a chemostat.

A comparison was made of acid production by cells of Streptococcus mutans Ingbritt and S. sanguis NCTC 7865 that had been washed twice and incubated in different concentrations of sodium and potassium ions. Organisms were grown under defined conditions in a chemostat under both glucose limitation and glucose excess conditions at a dilution rate of 0.1 h(-1) (mean generation time, 6.9 h). Acid production after a pulse of glucose, sucrose, and fructose was measured by pH fall experiments and as a rate at pH 7.0. S. mutans produced more acid than S. sanguis as measured by either criterion, although statistically faster rates of acid production and lower terminal pH values were obtained when cells of both species were suspended in KCl rather than in NaCl, with 200 mM KCl resulting in the lowest terminal pH in pH fall experiments. Sodium ions inhibited acid production: 183 mM NaCl reduced the glycolytic rates of S. mutans and S. sanguis metabolizing glucose at pH 7.0 in 135 mM KCl by 39 and 33%, respectively. The most pronounced stimulatory effect of potassium on acid production was by washed cells of S. sanguis that had been grown under arginine and under phosphate limitation. The pH fell by a further 0.86 and 1.21 pH units, respectively, and to below the critical pH for enamel demineralization when these cells were metabolizing glucose in 135 mM KCl compared with the same concentration of NaCl. This enhancement of acid production was not due to potassium translocation, as had been suggested previously, because no movement of potassium ions across the cell membrane could be detected. An alternative explanation is proposed in which sodium ions are excluded from the cell at the expense of membrane energy, i.e., the proton motive force, which could otherwise be used for the transport of sugars.

Fructose↗

Properties of Streptococcus mutans Ingbritt growing on limiting sucrose in a chemostat: repression of the phosphoenolpyruvate phosphotransferase transport system.

Growth of Streptococcus mutans Ingbritt on limiting sucrose in a chemostat at dilution rates of 0.05 to 0.4 h-1 (mean generation time, 14 to 1.7 h) resulted in a heterofermentative pattern of metabolic end products. During fast growth, lactic acid was the major end product, whereas at slower growth rates, acetic and formic acids, as well as ethanol, increased to be major end products. The patterns obtained were similar to those seen with the same organism growing on glucose. The glycolytic rate by washed cells was maximum at the lowest dilution rates and decreased as the cells were made to grow faster. Transport of sucrose, glucose, and fructose via the phosphoenolpyruvate phosphotransferase system (PTS) was repressed during growth on sucrose after growth on glucose. Uptake rates suggested that sucrose was transported in the PTS as the intact disaccharide. Comparison of the rate of sugar uptake in the chemostat with the rate of PTS activity in the cells at each growth rate indicated that the PTS was capable of supporting growth only at a dilution rate of 0.05 h-1. Growth on sucrose at faster growth rates required the activity of a second transport system, supporting our earlier observations with glucose that S. mutans contains at least two sugar transport systems.

Biological Transport↗