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Plasmid-determined resistance to erythromycin: comparison of strains of streptococcus faecalis and streptococcus pyogenes with regard to plasmid hmology and resistance inducibility.

Streptococcus faecalis strains DS-5 and Streptococcus pyogenes strain AC-1 both have a 17 million dalton plasmid that determines resistance to erythromycin, lincomycin, and vernamycin B(alpha). The results of deoxyribonucleic acid-deoxyribonucleic acid hybridization experiments indicate that the two plasmids are about 95% homologous. It was also shown that erythromycin resistance is inducible in AC-1 and constitutive in DS-5.

Drug Resistance, Microbial↗

Comparison of the post-antibiotic effect of Streptococcus faecalis and Streptococcus faecium with ampicillin alone or combined with streptomycin: studies on a novel type of antimicrobial interaction.

Determination of post-antibiotic effect (PAE) and time-kill studies were made with twelve strains of Streptococcus faecalis and nine strains of Streptococcus faecium, comparing the effect of ampicillin alone with a combination of ampicillin and streptomycin at achievable serum concentrations. Bactericidal synergism (greater than or equal to one log10 decrease in viable counts) and prolongation in PAE (greater than or equal to 0.5 h) were demonstrated in all streptomycin-susceptible strains (Minimum inhibitory Concentration less than 2000 mcg/ml), whereas only one of five highly streptomycin-resistant strains exhibited a synergistic effect. A significant correlation between the magnitude of increased killing and the increase in recovery period by the combinations of ampicillin and streptomycin was demonstrated.

Ampicillin↗

[Typing of Streptococcus faecalis and Streptococcus faecium strains with bacteriophages].

With a set of 17 bacteriophages specific for Streptococcus faecalis and a set of 7 bacteriophages specific for Streptococcus faecium 441 strains of S. faecalis and 53 strains of S. faecium were typed. The species identification was done by means of the API 20 STREP system. For S. faecalis 22 and for S. faecium 12 lysotypes were found. Among S. faecalis strains lysotype V6b was most common (27%), followed by the lysotypes XV (9,5%), XIII1 (7%), II1 (5,7%), and I1 (4,3%). These 5 lysotypes comprised 54% of all 441 strains. 21% of the strains of S. faecalis and 6 from 53 strains of S. faecium were untypable. Strains of S. faecium were most frequently isolated from faeces and in rare cases from urine. To the contrary 50% of the S. faecalis strains were isolated from urine. There was no correlation between special lysotypes and age of the patients or the source of specimen. The results of phage typing of several strains from one patient, and especially of strains from mothers and their newborns suggest the usefulness of the typing system to clarify epidemiological questions.

Adolescent↗

Activities of arginine dihydrolase and phosphatase in Streptococcus faecalis and Streptococcus faecium.

Strains of Streptococcus faecalis and S. faecium are known to produce ammonia from arginine, but only S. faecalis couples the adenosine triphosphate (ATP) produced through the arginine dihydrolase pathway to growth processes. The specific activities of the arginine dihydrolase enzymes were found to be much lower in S. faecium (0.01 to 0.10) than in S. faecalis (0.24 to 1.60). Phosphatase activities in both strains were similar (up to 0.11), but equaled or exceeded the activities of the arginine dihydrolase enzymes in S. faecium. The failure of S. faecium to show increased growth in arginine media is explained on the basis of low activities of the arginine dihydrolase enzymes coupled with sufficient phosphatase activity to negate any benefit from ATP formed.

Adenosine Triphosphatases↗

Streptococcus faecalis and Streptococcus faecium in infected dental root canals at filling and their susceptibility to azidocillin and some comparable antibiotics.

The study was performed on 612 routine cultures of material obtained from root canals of teeth at the time of filling (r-cultures) by students at the Department of Endodontics during a continuous period of one year. Twenty-nine isolates from 27 (29.3%) of the 92 positive cultures filled the criteria of enterococci (Sherman, 1937) and had demonstrable group D-antigen. With a set of tests these isolates were identified as follows: Strep. faecalis subsp. faecalis (10), Strep. faecalis subsp. zymogenes (3), Strep. faecalis subsp. liquefaciens (8), atypical variants of Strep. faecalis (6), Strep. faecium var. faecium (1) and Strep. faecium var. durans (1). Five tests in the present study clearly differentiated Strep. faecalis from Strep. faecium i.e. fermentation of sorbitol, glycerol (anaerobic) and melezitose, tolerance to potassium tellurite (0.1%) (positive for Strep. faecalis) and production of hydrogen peroxide (positive for Strep. faecium). In the inocula 10(3) or more colony forming units of enterococci were found more often of other identified microorganisms. This means that enterococci are of special interest in studies on the influence of infection at the time of filling of root canals on the prognosis of root canal therapy. The isolates were also tested for susceptibility to azidocillin, ampicillin, penicillin-G, penicillin-V and erythromycin with the paper disc method. All the isolates were susceptible to azidocillin and ampicillin (sensitivity group I), while the majority of the isolates showed a lower susceptibility to the other three antibiotics (sensitivity group II). The significance of these findings in the choice of prophylactic antibiotic to prevent bacterial endocarditis in patients with a history of rheumatic or congenital heart disease are discussed, when bacteremia from dental procedures may be expected.

Ampicillin↗

Isolation and structure of the bacterial sex pheromone, cAD1, that induces plasmid transfer in Streptococcus faecalis.

The Streptococcus faecalis sex pheromone cAD1, which is involved in the conjugative transfer of the hemolysin plasmid pAD1, has been isolated and its structure determined. Its Mr is 818 and its amino acid sequence is H-Leu-Phe-Ser-Leu-Val-Leu-Ala-Gly-OH. A replicate of the pheromone synthesized by the liquid-phase method showed the same biological activity and chromatographic behavior as the isolated cAD1. Pheromone activity was detectable at a concentration of approximately 5 X 10(-11) M.

Amino Acid Sequence↗

Characterization of two tetracycline resistance determinants in Streptococcus faecalis JH1.

Streptococcus faecalis strain JH1 harbors two conjugative plasmids: pJH1, an R plasmid mediating resistance to kanamycin, streptomycin, gentamicin, erythromycin, and tetracycline, and pJH2, a hemolysin-bacteriocin plasmid. Studies of plasmid-cured derivatives of strain JH1 and of transconjugates obtained after mixed incubation of JH1 with the plasmid-free S. faecalis strain JH2-2 revealed the presence of two tetracycline resistance determinants in strain JH1. One determinant mediated constitutive resistance to 40 micrograms of tetracycline per ml and was associated with plasmid pJH1. The second determinant, either on the chromosome of strain JH1 or on an undetectable plasmid, was inducible by tetracycline and enabled the host strain, in the absence of pJH1, to grow in the presence of 80 micrograms of tetracycline per ml. One transconjugant, strain DL172, was resistant to 80 micrograms of tetracycline per ml, sensitive to kanamycin, streptomycin, and erythromycin, and hemolytic in the presence, but not in the absence, of tetracycline. A single plasmid, pDL172, from this strain consisted of plasmid pJH2 and a 17.8-kilobase segment of DNA homologous to total cell DNA from strain JH1 but did not contain plasmid pJH1. Whether the addition of heterologous DNA to plasmid pJH2 occurred by translocation of a 17.8-kilobase tetracycline resistance transposon or by classical recombination with pJH2 has not been determined.

Base Sequence↗

Studies on Streptococcus faecalis enterotoxin.

Streptococcus faecalis has been reported to cause food poisoning. Six strains of S. faecalis were tested for Sherman's criteria. These strains were non-hemolytic, DNase+ and Ent+. The enterotoxin was purified on Sephadex G-200 column and maximum activity was observed at 37 C and pH 7.0. Enterotoxin treated with trypsin and papain elicited very poor response to fluid accumulation. The sensitivity of all the strains against different antibiotics was determined. Strain 53 M was treated with acridine orange and ethidium bromide and a total of 44 Amps Strr and 3 Amps Strs mutants were tested for toxin production. Out of these only 4 were toxin negative, amongst which 3 were also DNase negative and 1 showed partial DNase activity.

Acridine Orange↗

Autolytic enzyme system of Streptococcus faecalis. V. Nature of the autolysin-cell wall complex and its relationship to properties of the autolytic enzyme of Streptococcus faecalis.

Cell walls from exponential-phase cultures of Streptococcus faecalis ATCC 9790 contain an autolysin (a beta-N-acetylmuramide glycanhydrolase, E.C. 3.2.1.17) which has been isolated from trypsin-speeded wall autolysates. The autolysin, which was excluded from Bio-Gel P-60, was further fractionated by diethylaminoethyl (DEAE)-cellulose chromatography or filtration on Bio-Gel P-200. After DEAE-cellulose chromatography, which removed most of the wall polysaccharide, autolysin activity was extremely labile and was rapidly lost at -20 C, even in the presence of albumin. The P-60-excluded enzyme was rapidly bound by walls at both 37 C (50% bound in about 1 min) and 0 C (50% bound in less than 4 min). Wall-bound autolysin could not be removed by 1.0 m ammonium acetate (pH 6.9). Autolysin was also bound by walls that had been extracted with 10% trichloroacetic acid or treated with 0.01 n periodate, suggesting that the nonpeptidoglycan wall polymers are not important for binding. Wall-bound autolysin was more stable than the soluble enzyme to proteinase digestion, acetone (40%), 8 m urea (at 0 C), or to inactivation at 56 C. Two bacterial neutral proteinases (which do not hydrolyze ester bonds) activated latent wall-bound autolysin, suggesting that activation results from the cleavage of one or more peptide bonds. The group A streptococcal proteinase activated latent autolysin but differed from the other proteinases in that it did not inactivate soluble autolysin. The results suggest that the autolysin is not covalently linked to the wall. The high affinity of the walls for the autolysin appears to be responsible for the firm, not easily reversed binding.

Autolysis↗

Glycerophosphoryl phosphatidyl kojibiosyl diacylglycerol, a novel phosphoglucolipid from Streptococcus faecalis.

1. From Streptococcus faecalis a third novel phosphoglucolipid was isolated. It contained D-glucose, glycerol, acyl groups and phosphorus in a molar ratio of approx. 2 : 3 : 4 : 2. 2. The structure was shown to be 3(1)-O-[6 inches-(sn-glycerol-1-phosphoryl), 6 foot-(1,2 diacyl-sn-glycerol-3-phosphoryl)-2 foot-O-(alpha-D-glucopyranosyl)-alpha-D-glucopyranosyl]-1(3),2-diacylglycerol. The novel lipid is related to the earlier described phosphatidyl and glycerophosphoryl kojibiosyl diacylglycerol from Streptococcus faecalis by combininb their substituents in its structure. 3. The structure of the deacylated core was accomplished by analyses of the breakdown products obtained on Smith degradation and strong alkaline hydrolysis. The location of the acyl groups was achieved by degradation of the intact lipid with 60% HF (w/v) which resulted in the formation of inorganic phosphate, glycerol, diacylglycerol and diglucosyl diacylglycerol. The sn-glycerol-1-phosphoryl and 3-sn-phosphatidyl substituents were located by hydrolysis of the lipid with 98% acetic acid (v/v) and subsequent analysis of the resultant phosphatidyl diglucosyl diacylglycerol. 4. The composition and positional distribution of the constituent fatty acids at the two diacylglycerol portions was studied by combination of chemical and enzymatic degradations. The fatty acids are the same as with the other polar lipids of S. faecalis, they are present in nearly the same proportions and display the same non-random distribution pattern. 5. A possible relationship between sn-glycerol 1-phosphate containing phosphoglucolipids and lipoteichoic acid will be discussed.

Chromatography, Gas↗

Differentiation of Streptococcus faecalis Andrewes and Horder and Streptococcus faecium Orla-Jensen based on the amino acid composition of their murein.

The amino acid composition of the murein (peptidoglycan) of 75 strains of enterococci was investigated. All strains designated Streptococcus faecalis according to their physiological properties contained the lysine-alanine type of murein; all strains classified as S. faecium contained the lysine-aspartic acid type of murein.

Amino Acids↗

Synthesis of catalase by "Streptococcus faecalis subsp. zymogenes".

"Streptococcus faecalis subsp. zymogenes" was grown aerobically and anaerobically with glycerol as the source of carbon, and in the presence and absence of haematin. Catalase activity was found only during aerobic growth in the presence of haematin. The rate of appearance of catalase activity was measured on (a) addition of haematin to haematin-less aerobic cultures, and (b) aeration of haematin-containing anaerobic cultures in the presence or absence of chloramphenicol. These experiments suggested that apocatalase synthesis was induced by aeration and was not dependent on the presence of haematin in the growth medium. The binding of haem to apocatalase was oxygen dependent.

Aerobiosis↗

Factors influencing the frequency of mesosomes observed in fixed and unfixed cells of Streptococcus faecalis.

Mesosomes of Streptococcus faecalis (American Type Culture Collection 9790) were seen about 92% less frequently in freeze fractures of unfixed cells than in freeze fractures and sections of fixed cells. This difference in frequency was not related to any period of unbalanced macromolecular synthesis induced by chemical fixation. All measured synthetic processes (DNA, RNA, and protein synthesis, and glycerol incorporation) were halted with either osmium tetroxide (OS) or glutaraldehyde fixation. That fewer mesosomes were seen in freeze fractures of unfixed cells was probably due to the difficulty of observing cross-fractured mesosomes in this organism in the unfixed state. Unfortunately, mesosomes probably preferentially cross fracture in the unfixed state and therefore are usually only observed, infrequently, in those cases where the freeze fracture follows the surface layer of a mesosomal membrane. However, the addition of glycerol to unfixed cells, especially in the chilled state, greatly increased the frequency of observation of cytoplasmic mesosomes in freeze fractures. It is thought that glycerol, like chemical fixation, increases the number of surface-fractured mesosomes, which in turn increases the frequency of mesosome observation. It was also observed that cellular autolysis occurring during OS fixation seemingly reduced the number of mesosomes observed in thin sections and freeze fractures of OS-fixed cells.

Autolysis↗