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The effects of variations in growth conditions on the penicillin response of Streptococcus faecium ATCC 9790 were studied. Changes in the growth temperature and medium composition were found to cause striking changes in the bacterial generation time, cellular penicillin sensitivity (minimum inhibitory concentration), sensitivity of peptidoglycan synthesis to inhibition by penicillin, rate of autolysis, and labeling pattern of penicillin-binding proteins. However, no constant relationship between these parameters and the minimum inhibitory concentration could be observed. Similar electrophoretic patterns for penicillin-binding proteins were observed in cells grown in different media at the optimal growth temperature. Inhibition of cell division by penicillin in cells grown at this temperature (but not at higher or lower temperatures) caused filamentation of the bacteria. In cells grown in a chemically defined medium at the optimal temperature (but not at temperatures above or below), complete inhibition of cell division was associated with only partial inhibition (34% after 150 min) of peptidoglycan synthesis. It is suggested that the status and physiological importance of individual penicillin-binding proteins in S. faecium are heavily influenced by growth conditions. Depending on the growth conditions, different penicillin-binding proteins may perform the cellular function, indispensible for bacterial growth.

A number of purified cell walls of various gram-positive bacteria had arthritogenic activity in the rat. The water-soluble adjuvant-active component(s), which were isolated from some of these cell walls by utilizing a peptidoglycan-degrading enzyme, did produce severe adjuvant arthritis. However, the components obtained by digestion with glycan-degrading enzymes failed to produce arthritis. Thus, the present finding indicates the importance of a peptidoglycan portion, especially its intact glycan chain, for induction of adjuvant arthritis.

The authors have developed a method permitting the microscopic study of the morphology of bacteria and their relative position in microcolonies. Thus, the use of this method has made it possible to distinguish Streptococcus pneumoniae from other bacteria by the morphology of their microcolonies. In the study of 75 streptococcal strains, all strains yielding positive results in two or three tests, similarly to all strains pathogenic for mice, formed microcolonies with granular (pneumococcal) morphology, while all strains yielding negative results have been found to form microcolonies with catenulate ("nonpneumococcal") morphology. The authors suggest that the morphology of microcolonies is a more reliable criterion for the identification of S. pneumoniae than other tests used separately.

Isolated were a total of 33 Streptococcus strains during two enzootics in adult and young chickens. Their cultural, morphologic, and biochemical aspects were studied in order to elucidate the biotype to which they belonged. Taking into consideration their capacity to ferment trehalose and the fact that they failed to ferment sorbite along with their beta-hemolytic activity, belonging to serologic group C, these Streptococcus cultures were determined as Streptococcus pyogenes hum. C.

Segregation was studied by measuring the positions of autoradiographic grain clusters in chains formed from single cells containing on average less than one radiolabeled chromosome strand. The degree to which chromosomal and cell wall material cosegregated was quantified by using the methods of S. Cooper and M. Weinberger, dividing the number of chains labeled at the middle. This analysis indicated that in contrast to chromosomal segregation in Escherichia coli and, in some studies, to that in gram-positive rods, chromosomal segregation in Streptococcus faecium was slightly nonrandom and did not vary with growth rate. Results were not significantly affected by strand exchange. In contrast, labeled cell wall segregated predominantly nonrandomly.

The same tendency that causes soap bubbles to achieve a minimum surface area for the volume enclosed seems to account for many of the features of growth and division of bacteria, including both bacilli and cocci. It is only necessary to assume that growth takes place in zones and that only in these zones does the tension caused by hydrostatic pressure create the strain that forces the cell to increase the wall area. The stress developed by osmotic pressure creates strains that significantly lower the free energy of bond splitting by hydrolysis or transfer. We believe this is sufficient to make growing wall have some of the properties ordinarily associated with surface tension. The feature common to all bacterial cell wall growth is that peptidoglycan is inserted under strain-free conditions. Only after the covalent links have been formed are the intervening stressed peptide bonds cleaved so that the new unit supports the stress due to hydrostatic pressure. The present paper analyses the growth of Streptococcus faecium in these terms. This is a particularly simple case and detailed data concerning morphology are available. The best fit to the data is achieved by assuming that growth takes place in a narrow region near the splitting septum and that the septal material is already under tension as it is externalized and is twice as thick as the external wall throughout the development of the nascent poles. Constancy of the ratio of hydrostatic pressure to the effective surface tension, P/T, is also consistent with electron microscopic observations.

Contact angle measurements on layers of bacteria were used to determine the bacterial surface free energy (gamma b) of a variety of oral streptococcal strains, both without and after being coated with human whole saliva. At least four isolates of each species, either freshly isolated or laboratory strains, were used. The species Streptococcus mutans, S. sanguis, and S. salivarius were homogeneous, having high surface free energies, and were not affected by saliva treatment (gamma b = 106 +/- 12 and 107 +/- 10 erg X cm-2 in the absence and presence of saliva coating, respectively; n = 20). S. mitis had a very low surface free energy (46 +/- 15; n = 5), which was significantly increased after salivary adsorption (71 +/- 14 erg X cm-2; p less than 0.002). The species S. milleri contained strains with both high and low gamma b. Calculation of the interfacial free energy of adhesion (delta F adh) for bacteria from a saliva suspension to solid surfaces with various arbitrary surface free energies (gamma s) showed that, theoretically, most strains will encounter thermodynamically favorable conditions for adhesion to surfaces with a gamma s above 62 erg X cm-2. However, S. mitis strains not coated with saliva would only be able to adhere to surfaces with gamma s lower than this value. Saliva-coating reverses the calculated relationship with gamma s for these strains. The results indicate that an enamel surface with a low gamma s value would be thermodynamically unfavorable for adhesion of most oral streptococci.

Some, but not all strains of Streptococcus salivarius were demonstrated to occur in smooth (S) and rough (R) variants, growing in distinctly different colonies on sucrose-containing agar plates. Sucrose-derived extracellular polysaccharides (EP) of NCTC 8606, ATCC 9759 S and R, ATCC 13419 S and R, 1A S and R, and Tove S and R were isolated, purified, and chemically studied. Extracellular enzymes of R variants yielded more water-insoluble than soluble material, while the opposite was true for S variants. The insoluble material consisted mainly of glucan, the soluble mainly of levan. Enzymatic hydrolysis suggested a predominance of alpha-1,3-linkages in the water-insoluble glucan. Cell-associated enzymes gave rise to cell-associated and -free EP. The cell-associated EP of S variants was insoluble, while that of R variants contained water-insoluble glucan and water-soluble fructan. Cells coated with cell-associated EP flocculated due to interaction of the EP, mainly by hydrogen bonding, in part by divalent cation bridging. The sucrose-derived EP gave rise to plaque deposit formation in sucrose broth cultures, S variant deposits being thin and firmly adherent to glass, R variant deposits being thick, rough, coherent, but only weakly adherent. The variant types were not altered by the curing agents ethidium bromide and acridine orange.

The microflora from the Armenian yoghurt was studied during its growth on the optimal for the medium (skim milk) using objectives of the Peshkoff system. It was shown that certain organisms of Lactobacterium jogorti tended to branch at the early stages of their growth, and the cells elongated and formed complex coils and helices by the third-fourth day of growth. Compact aggregates of cocci having angular outlines appeared in the culture of Streptococcus thermophilus by the end of the second day of growth. Lysis of individual organisms, and occasionally zones of lysis, were found in the both cultures at certain stages of their growth. Cultures grown for 4-5 days were characterized by wide polymorphism.

The buoyant density of whole cells of Streptococcus faecium varies with growth rate and during the cell cycle. Two possible explanations for this were explored: (i) the density of cell walls may vary, and (ii) the proportions of wall and cytoplasm may vary. We tested the first possibility by isolating walls from chilled, unfixed populations of S. faecium cells and fractionating them on Percoll density gradients. Mean cell wall density averaged 4% less than whole-cell density and did not vary significantly with growth rate. In addition, walls isolated from heavy and light fractions of a population of cells did not differ significantly in density. Thus, variation in the density of isolated cell walls could not account for the observed variation in whole-cell density within or between populations. Using previously published measurements of the physical dimensions of S. faecium cells, we also found that the relative proportions of wall and cytoplasm (see the second possibility above) could not account for the observed changes in whole-cell buoyant density.

Novel bacterial structures have been demonstrated in lysed blood filtrates placed in special culture media from patients with idiopathic hematuria. These structures converted rapidly to gram-positive coccal (streptococcal and staphylococcal-like) coccobacillary and filamentous, bacterial forms in vitro from 96 per cent of the patients studied. Blood cultured conventionally yielded negative findings. Although structures (dense bodies) were demonstrated in normal control blood specimens (albeit in lesser numbers) few converted to classical bacteria in vitro (7 per cent). Erythromycin therapy appeared to correlate with disappearance of hematuria and inability to revert rapidly the variant froms to classical bacteria in vitro. It is suggested that continual bombardment of the blood by bacteria entering from the mouth or other sites may lead to the development of variant bacterial parasitism. In an effort to survive the deleterious host effects the organisms may convert to persisting osmotically stable variant bacterial forms (dense bodies). Development of a disease state may be conditioned by some existing or developing abnormality in the host (immunologic, physiologic and/or biochemical). Furthermore, changes (genetic?) that might take place in the organisms per se during their transition to variant forms and adaptation to life in vivo may not allow certain host environments to adapt to these new forms, possibly leading to a pathogenetic role in renal diseases whose etiologies have long been enigmas.

Changes in the cell wall and various stages of lysis become manifested on the 1-2 day of development of the yogort bacteria. Heteromorphous individuals appear both in the culture of streptococci and rods. The nuclear apparatus of the yogort bacteria is exteremely polymorphous whereas in the streptococci it undergoes changes from compact nucleoids which are either round or crescent shaped or take the form of massive super-nuclei in heteromorphous individuals. A certain dynamics of fat and metachromatin (volutin) was noted in the process of the development of the culture. The presence of correlation was shown among the maximal fermentative activity of the culture, the beginning of lysis, the appearance of heteromorphous forms and volutin accumulation.

One of the most important questions in ecology is how to explain the co-existence of the variety of physiologically related organisms in the same habitat. A model is presented for the co-existence of Streptococcus species and Actinomyces species in dental plaque. The hypothesis is that these organisms co-exist because they simultaneously utilize several carbon and energy substrates. The hypothesis follows from the observation that the growth yield of oral streptococci and actinomyces in saliva is limited by carbohydrate. Preliminary experiments were undertaken to test the hypothesis using mixed chemostat cultures and gnotobiotic rats. Competition between S. mutans K1R and A. viscosus Ut2 in mixed chemostat cultures on glucose and asparagine was hampered by the early appearance of high-glucose-affinity variants of A. viscosus. From the physiological characteristics of S. sanguis and S. milleri, it might be predicted that simultaneous utilization of carbohydrate and arginine would enable these organisms to co-exist with S. mutans in an ecosystem. To test this mechanism under natural conditions, germ-free rats were inoculated with a combination of S. mutans K1R and S. sanguis P4A7 or the combination S. mutans K1R and S. milleri B448. The rats were fed on three different diets: (1) 58% cornstarch; (2) 48% cornstarch and 10% sucrose; and (3) 53% cornstarch and 5% arginine. The results of this experiment demonstrated that dietary arginine caused a significant decrease of the ratios K1R/P4A7 and K1R/B448 in dental plaque.(ABSTRACT TRUNCATED AT 250 WORDS)

The author summarizes research performed by himself and his colleagues to study chronic erosive arthritis by means of injecting fragments of streptococcal cell walls into rats. By study of rat strains resistant and susceptible to various stages of the disease, as well as the effects of therapeutic drugs, information has been obtained on the pathogenesis of the synovitis and on the role of cellular and humoral factors in regulating inflammatory disease.

Isolation of colonies with altered colonial morphology was frequently observed following infection of specific pathogen-free rats fed a caries-inducing high sucrose diet with two human strains of Streptococcus salivarius. These isolates produced unique rough colonies on mitis salivarius agar, in contrast to the smooth, mucoid colonial morphology of the parental strain. The rough variants were characterized by a marked increase in the synthesis of polysaccharide from sucrose, especially cell-associated water-insoluble glucan, whereas the smooth parental strains mainly produced cell-free fructan. Glucose-grown cells of rough variants but not smooth parental strains were aggregated upon addition of sucrose. Such variants were not detected during laboratory transfers of the human strains of S. salivarius nor by mutagenesis of the cultures.

The incidence of S. sanguis, S. salivarius, S. mutans, total streptococci and lactobacilli was examined in highly caries active 13-14-year-old schoolchildren participating in a prophylactic program. After 1 year of trial, professional toothcleaning once every second week markedly reduced the frequency of gingivitis and the caries increment. Bimonthly topical applications of a 0.5% chlorhexidine gel with or without subsequent rinsing with 2% NFP had no effect on plaque score and gingivitis but tended to reduce the caries activity. No significant changes were found in the groups with regard to the salivary number of total streptococci and S. sanguis. A reduction of the population of S. salivarius, S. mutans and lactobacilli was observed in the chlorhexidine group. In the group where chlorhexidine was combined with MFP, only S. mutans was reduced.

The regrowth of 27 temperature-sensitive division mutants of Streptococcus faecium ATCC 9790 was examined after various periods of incubation at the nonpermissive temperature. Several of the mutants blocked at various stages of septum formation or of daughter-cell separation divided in a partially or completely synchronous way after a short incubation at the nonpermissive temperature. All four lytic mutants blocked early in the cell division cycle divided at a normal rate after a brief lag.

Glucose or sucrose grown cells of Streptococcus mitis ATCC 903 bind spontaneously to the surface of each other producing visible microbial aggregates upon incubation in 10 mM phosphate, citrate-phosphate or tris-maleate buffers. Aggregation was delayed and proceeded at a slower rate when bacteria grown in a culture medium with a low carbohydrate/nitrogen ratio were used. Growth in this culture medium resulted in carbohydrate limitation. The aggregation was highly reproducible and was unaffected by pH in the range of 4.4-7.0 but was decreased at pH 8.0 and completely inhibited at pH 9.0. No inhibition of the reaction was observed when a series of simple and complex carbohydrates were added. There was no significant difference in the rate of aggregation at 20, 30 and 37 degrees C. Aggregation occurred at a demonstrable rate of 0 degrees. Chloramphenicol did not inhibit aggregation. Since inhibition of aggregation was obtained by treatment of bacteria with trypsin or heat it appears that protein of glycoprotein components on the bacterial surface were involved in the reaction.

A motile Streptococcus was isolated and its chemotactic behavior toward sugars and amino acids was studied. Motility was optimal in the presence of an exogenous energy source and a nonionic detergent, e.g., Tween 80 or Brij-36. Both glucose and pyruvate could serve as energy source. Chemotaxis toward leucine was optimal at pH 7 to 8.5 and a temperature between 30 and 37 C. The Streptococcus showed a chemotactic response toward a variety of sugars. All commonly occurring L-amino acids, except alanine, asparagine, aspartate, glutamate, arginine, and lysine, were attractants. From concentration response curves the thresholds, peak concentrations, and optimal responses were determined.

There is a high morbidity and mortality associated with brain abscesses in children with congenital cyanotic heart disease. A case is reported here which implicated an endodontically treated primary molar in the etiology of a brain abscess in a boy with congenital cyanotic heart disease.

Bacteria of the Lactococcus (formerly lactic Streptococcus) genus are described to emphasize certain less well-known facts of importance in their successful use as dairy starters. Other streptococci (non-rod forms) are briefly characterized to emphasize their usefulness in industrial milk fermentations.

A sensitive, in vitro method has been developed for the study of bacterial adsorption to dental crown and bridge cements. The method employs heavily radiolabelled cells which adhere to the wall in wells made from the cements. Loss of radioactivity from the bacterial suspension during incubation is used to measure the degree of bacterial adherence to the materials. Strains of Streptococcus sanguis and Streptococcus mutans adhered in greater numbers than a strain of Streptococcus salivarius to both a zinc phosphate cement and a zinc carboxylate cement. The zinc phosphate cement supported adherence to a greater degree than did the zinc carboxylate cement. Pretreatment of the cement surfaces with human saliva tended to reduce bacterial adherence. Pretreatment of the bacteria with saliva slightly enhanced sorption of S. sanguis and S. salivarius, but tended to impair sorption of S. mutans to both types of cements. When both bacteria and cements had been pretreated with saliva, a strong inhibition of the adherence of S. mutans was observed.

The metabolism of aflatoxin B1 to other fluorescent compounds was studied in rumen organisms. No fluorescent metabolites were found by chromatographic procedures which separate aflatoxin B1 from them. Rumen bacteria (mixtures and Streptococcus bovis) incubated with aflatoxin B1 grew in long chains whereas cells in control cultures were mainly doublets with some short chains. Growth of mixed rumen bacteria and Streptococcus bovis cultures was inhibited by aflatoxin B1 as measured by change in optical density and by incorporation of [carbon-14] adenine into nucleic acids.

A method is developed for cultivating and observing bacteria in an early phase of their growth, when microcolonies were formed. The morphology of the microcolonies, including form, structure, characteristics of its periphery and center, as well as the mode of arrangement of bacteria in it and to each other proved to be typical for a species and often permitted its differentiation from the other species. Photographs are presented of typical microcolonies of S. aureus and of E. coli. A series of photographs is presented also as an illustration of the possibility to differentiate some species in the genus Streptococcus. The microcolonies observation is made 3 hours after material inoculation, that may permit a rapid bacteriological diagnosis. It is believed also, that the microcolony technique could be useful in the characterisation and identification of the species in the general bacteriology and taxonomy.

Experimentally induced plaque seemed to originate by direct contact inoculation from the vestibular mucosa and saliva. During the next seven days, this plaque developed its own characteristics. Populations of Streptococcus mutans usually less than 2% of total streptococci population in plaques that were less than three days old, increased between days 3 and 7. Proportions of S sanguis, high in early samples, decreased after day 3. Populations of S salivarius, which usually outnumbered other streptococci, fluctuated widely through day 3, and then increased in proportion in subjects who were more productive of experimental caries, but decreased in subjects who were less productive. Proportions of plaque flora comprising lactobacilli paralleled those of S salivarius.

This study was designed to determine whether poly-HEMA root canal filling material can support bacterial growth. Eight different microorganisms were tested against the polymerized material, as well as against the powder and paste components of the unpolymerized material. The results establish that this root canal filling material does not support bacterial growth.

A prophylaxis followed by three topical applications of an iodine-potassium iodide solution significantly reduced the levels of Streptococcus mutans in fissure and approximal plaques and in saliva. Reductions persisted 20--24 weeks after treatment in salivary and approximal samples. A prophylaxis alone exerted a small and temporary reduction of S. mutans in occlusal fissure plaque, but did not reduce the levels of this organism in approximal plaque or in saliva. A significant relationship existed between the levels of S. mutans in saliva and the proportions of this organism in plaque. The dorsum of the tongue does not appear to constitute a significant reservoir for S. mutans following disinfecting procedures.