Search

Background/aims: Microorganisms of Peptostreptococcus micros are asaccharolytic, anaerobic gram-positive cocci that are frequently isolated from human oral sites such as periodontal pockets. Preliminary study showed that several amino acids, including serine, enhanced slightly the growth of P. micros. Therefore, we investigated the degradation of serine and serine-containing oligopeptides., Methods: Metabolic end products were determined with high-performance liquid chromatography. The related enzymatic activities in cell-free extract were also assayed., Results: Washed P. micros degraded serine-tripeptides (Ser-Ser-Ser), and produced formate, pyruvate, acetate, and ammonia. They also degraded serinyl-tyrosine (Ser-Tyr) to the same products. Related enzymatic activities, such as serine dehydratase, pyruvate formate-lyase, formate dehydrogenase, pyruvate oxidoreductase, phosphate acetyltransferase, and acetate kinase, were detected in the cell-free extract, indicating that the organisms produced ATP in the serine metabolism., Conclusion: P. micros utilized serine-containing oligopeptides as exogenous metabolic substrates rather than serine itself, and degraded Ser-Ser-Ser and Ser-Tyr to formate, pyruvate, acetate, and ammonia with ATP generation.

Peptostreptococcus micros is a gram-positive bacterium that has been associated with periodontitis and endodontic infections. In this study, we hypothesized that P. micros binds the immunomodulating component lipopolysaccharide derived from gram-negative bacteria to increase its capacity to stimulate cytokine production by host cells. The ability of P. micros to bind Actinobacillus actinomycetemcomitans lipopolysaccharide was demonstrated by an enzyme-linked immunosorbent assay and by immunoelectron microscopy. Pretreatment of P. micros cells with A. actinomycetemcomitans lipopolysaccharide was associated with a 49-fold increase in tumor necrosis factor alpha production by human monocytic cells U937 differentiated into adherent macrophages, compared to the stimulation with untreated P. micros. This effect was suppressed by incorporating polymyxin B, a lipid A-binding substance, during treatment of macrophage-like cells with lipopolysaccharide-coated P. micros cells. This is the first study reporting a binding interaction between lipopolysaccharide and a gram-positive bacterium. This interaction represents a new mechanism that could promote the inflammatory response during periodontitis.

Peptostreptococcus micros, an anaerobic gram-positive coccus, has been associated with periodontal and endodontic lesions, including those refractory to treatment, as well as many human polymicrobial infections in other body locations. A selective and differential medium for the primary isolation of P. micros was developed and evaluated. Columbia CNA agar, a selective medium for gram-positive cocci, was supplemented with glutathione and lead acetate (P. micros medium: PMM). P. micros has a characteristic of rapidly utilizing the reduced form of glutathione to form hydrogen sulfide, which reacts with lead acetate producing a black precipitate in the medium. When grown on PMM, P. micros can be easily identified by its typical colonial morphology and the presence of a black precipitate directly under the colony. PMM was compared for the growth of P. micros with phenylethyl alcohol agar (PEA) and Columbia base medium (CBM) with 80 strains of P. micros and 30 strains of other gram-positive cocci. All P. micros isolates tested grew and showed the typical morphology of P. micros on PMM. Using colony counts on CBM as controls, there was an average 81.8% recovery in the number of P. micros colonies on PMM, in contrast to an average 6.1% recovery on PEA. Subgingival plaque and tongue samples from 12 adult periodontitis and 6 early-onset periodontitis patients were cultured onto PMM for the isolation of P. micros. P. micros was isolated on PMM and identified biochemically and enzymatically from both adult periodontitis and early-onset periodontitis patients with higher percentages isolated from the diseased periodontal pockets of adult periodontitis patients; furthermore, this is the first isolation of P. micros from tongue samples taken from periodontally diseased patients. This medium in cultural studies will further our understanding and assist future investigations of P. micros involved in disease processes.

There are high amounts of hydrogen sulfide in deep periodontal pockets. This volatile sulfur compound may be formed from L-cysteine, but only low levels of this amino acid can be expected to be present in periodontal pockets. Glutathione, L-gamma-glutamyl-L-cysteinylglycine, is in high concentration in most tissue cells, and this tripeptide may be more readily available as a source of hydrogen sulfide formation in the pockets. The ability of 37 different species of oral bacteria to utilize glutathione in hydrogen sulfide formation was studied. Of these species, only 2 species of Peptostreptococcus and 5 species of Fusobacterium formed high amounts of hydrogen sulfide from glutathione within 24 h. Since the initial rate of hydrogen sulfide formation was more than 5 times higher in Peptostreptococcus micros than in any of the other bacterial species, the kinetics of sulfide formation from glutathione by P. micros was further elucidated. The formation of sulfide followed quite closely hyperbolic Michaelis-Menten kinetics. The maximal initial rate of sulfide formation (Vmax) was 163 +/- 2 nmol sulfide per minute per milligram of cellular protein. Half maximal initial rate (Km) was obtained at 7.4 +/- 0.8 microM glutathione. The initial rate of sulfide formation from L-cysteine was much slower and was almost proportional to L-cysteine concentration. This difference in kinetics of sulfide formation between glutathione and L-cysteine strongly suggested that glutathione was actively transported into the cell, whereas the transport of L-cysteine was more or less controlled by diffusion. The sulfide formation from the dipeptide L-cysteinylglycine also followed quite closely hyperbolic Michaelis-Menten kinetics.(ABSTRACT TRUNCATED AT 250 WORDS)

Lactobacilli from women with and without bacterial vaginosis (BV) were tested for H2O2 production. Thirty-seven (79%) of the 47 strains of lactobacilli isolated from the women without BV produced H2O2, while only nine (23%) of the 39 strains of lactobacilli obtained from women with BV did so. Five of 20 H2O2-producing and two of 26 non-producing strains of Lactobacillus exhibited antibiosis against four of 12 strains of peptostreptococci and two of 10 strains of Mobiluncus spp. None of a further 41 different anaerobic and facultative anaerobic bacterial strains were inhibited by any of the isolates of lactobacilli tested. Some strains of Gardnerella vaginalis, Bacteriodes spp., Mobiluncus spp. and Peptostreptococcus spp. inhibited the growth of three strains of lactobacilli belonging to different species. When the pH of the culture medium was increased from 6.0 to 6.5 this led to a decrease in the number of strains inhibited and/or the size of the growth-inhibitory zones. Different concentrations of H2O2 did not inhibit any of the strains tested. The growth-inhibitory effect of lactobacilli could not be related to their bacteriocin production. Increasing the iron content of the medium by adding FeCl3 (0.01 mM-1 mM) decreased or completely abolished the antibiosis.

Water-soluble intestinal mucins were investigated in germfree (GF), exgermfree (EXG) and conventional (CONV) rats and in GF and CONV mice. After agar gel electrophoresis, all GF animals had similar specific band patterns demonstrated by PAS and Toluidine Blue. These patterns, never seen in CONV animals, disappeared in GF animals infected either with intestinal contents from CONV rats or mono-infected with a mucin converting microorganism, labelled Peptostreptococcus N. The intestinal microflora seem to have a profound influence on the water-soluble mucins, and specific microorganisms appear to be involved in the conversion of these substances. Any CONV animal with a GF mucin pattern in the faeces must be considered to have a disturbance of the normal intestinal microflora.

A mucin-degrading microorganism was isolated from the intestinal tract by serial sectioning from the serosal side of the caecum wall from a conventional rat. The ability of degrading the intestinal water-soluble mucin was present both in vivo after monocontamination of germ-free rats and in vitro, when adding the microbe to Mucin medium. The morphology, Gram-positive cocci single or in short chains and the very weak biochemical activities allow us to place this strain in the species Peptostreptococcus micros.