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Serologic studies of Streptococcus intermedius, Streptococcus constellatus, and Streptococcus morbillorum by crossed immunoelectrophoresis.

A reference antigen-antibody system for Streptococcus intermedius, Streptococcus constellatus, and Streptococcus morbillorum was established with crossed immunoelectrophoresis. A comparison of S. intermedius, S. constellatus, and S. morbillorum with crossed immunoelectrophoresis and crossed immunoelectrophoresis with intermediate gel indicated that S. intermedius and S. constellatus are closely related antigenically with as many as six common cytoplasmic antigens. S. morbillorum was antigenically more distinct; antiserum of one strain of S. morbillorum was monospecific, indicating that specific serogroups of S. morbillorum exist. Crossed immunoelectrophoresis and tandem crossed immunoelectrophoresis revealed that S. intermedius, S. constellatus, and S. morbillorum also share some common antigens with Streptococcus sanguis and Streptococcus mitis, but S. intermedius, S. constellatus, and S. morbillorum are antigenically distinct from Streptococcus mutans and Streptococcus bovis.

Antibodies, Bacterial↗

Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus ("Streptococcus milleri group") are of different clinical importance and are not equally associated with abscess.

Difficulties in distinguishing organisms of the "Streptococcus milleri group" (SMG; Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus), have caused ambiguity in determining their pathogenic potential. We reviewed 118 cases in which SMG isolates had been identified using 16S rDNA sequence. S. constellatus and S. anginosus were isolated far more frequently than was S. intermedius. Nearly all isolates of S. intermedius and most isolates of S. constellatus, but only 19% of those of S. anginosus, were associated with abscess. Our findings suggest that speciation of the SMG may guide diagnostic evaluation, give insight into the possible role of coinfecting organisms, and help assess the need to search for occult abscess.

Abscess↗

Phenotypic differentiation of Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus strains within the "Streptococcus milleri group".

A biochemical scheme was developed by which strains of Streptococcus constellatus, Streptococcus intermedius, and Streptococcus anginosus can reliably be distinguished from within the "Streptococcus milleri group." Strains identified as S. intermedius were differentiated by the ability to produce detectable levels of alpha-glucosidase, beta-galactosidase, beta-D-fucosidase, beta-N-acetylgalactosaminidase, beta-N-acetylglucosaminidase, and sialidase with 4-methylumbelliferyl-linked fluorogenic substrates in microdilution trays after 3 h of incubation at 37 degrees C, together with the production of hyaluronidase. Strains of S. constellatus and S. anginosus were differentiated by the production of alpha-glucosidase and hyaluronidase by the former and the production of beta-glucosidase by the latter. The majority of strains of the S. milleri group obtained from dental plaque were identified as S. intermedius, as were most strains isolated from abscesses of the brain and liver. Strains of S. constellatus and S. anginosus were from a wider variety of infections, both oral and nonoral, than were strains of S. intermedius, with the majority of strains from urogenital infections being identified as S. anginosus.

Dental Plaque↗

Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus (the Streptococcus milleri group): association with different body sites and clinical infections.

The associations of Streptococcus intermedius, S. constellatus, and S. anginosus (the three species of the S. milleri group) with clinical infections and sites of isolation were investigated by using a simple biochemical scheme to identify a collection of 153 clinical isolates. S. intermedius was associated with abscesses of the brain and liver, while both S. anginosus and S. constellatus were isolated from a wider range of sites and infections. S. anginosus strains predominated in both genitourinary and gastrointestinal sources and exhibited a wider range of phenotypes, particularly in the ability to ferment mannitol and/or raffinose.

Central Nervous System Diseases↗

Effect on polymorphonuclear cell function of a human-specific cytotoxin, intermedilysin, expressed by Streptococcus intermedius.

Streptococcus intermedius is a member of the normal flora of the mouth but is also an opportunistic pathogen associated with purulent infections at oral and nonoral sites. Intermedilysin (ILY) has been shown to be a cytolysin capable of generating pores in the cell membrane of erythrocytes demonstrable by electron microscopy. This effect has been shown to be specific for human cells. Since polymorphonuclear cells (PMNs) are the main cell involved in innate immunity we investigated the effect of purified intermedilysin from Streptococcus intermedius on PMN function. Active ILY at a concentration of 40 ng/microl caused a significant decrease in the number of intact PMNs after 60 min. The active cytolysin, when compared with heat-inactivated ILY, did not appear to be chemotactic for the PMNs but did cause an increase in intracellular calcium, with increased cell surface CD11b expression, metabolic burst, and phagocytosis of Staphylococcus aureus. These findings may have implications for the role of ILY in deep-seated abscesses.

Bacterial Proteins↗

Emended descriptions and recognition of Streptococcus constellatus, Streptococcus intermedius, and Streptococcus anginosus as distinct species.

Strains currently classified as Streptococcus anginosus include strains previously identified as Streptococcus constellatus (Prevot 1924) Holdeman and Moore 1974, Streptococcus intermedius (Prevot 1925), and "Streptococcus milleri" (Guthof 1956) because these specific epithets were argued to be later synonyms of Streptococcus anginosus (Andrewes and Horder 1906) Smith and Sherman 1938 by Coykendall et al. (Int. J. Syst. Bacteriol. 37:222-228, 1987). However, recent data from DNA-DNA hybridization experiments, whole-cell-derived polypeptide patterns determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and data from phenotypic testing have demonstrated that Streptococcus anginosus strains represent three readily identifiable taxa to which the previously assigned type strains of Streptococcus constellatus (strain NCDO 2226 [= ATCC 27823], Streptococcus intermedius (strain NCDO 2227 [= ATCC 27335], and Streptococcus anginosus (strain NCTC 10713 [= ATCC 33397] have been shown to belong. Therefore, we propose recognition of Streptococcus constellatus (emend.) (type strain NCDO 2226 [= ATCC 27823]), Streptococcus intermedius (emend.) (type strain NCDO 2227 [= ATCC 27335]), and Streptococcus anginosus (emend.) (type strain NCTC 10713 [= ATCC 33397]) as distinct species and propose an emended description of each of these taxa.

Base Composition↗

Degradation and utilisation of chondroitin sulphate by Streptococcus intermedius.

Streptococcus intermedius, part of the 'Streptococcus milleri group', has the ability to produce glycosaminoglycan depolymerising enzymes (hyaluronidase and chrondroitin sulphate depolymerase) which is unique amongst the viridans streptococci and may contribute to their virulence in brain and liver abscesses. The growth of S. intermedius strain UNS 35 was studied in basal medium supplemented with chondroitin sulphate A (CS-A, sulphated at position 4 of the N-acetylgalactosamine moiety) or chondroitin sulphate C (CS-C, sulphated at position 6 of the N-acetylgalactosamine moiety) as the major carbohydrate source. CS-A but not CS-C supported the growth of S. intermedius. Extracellular degradation of CS-A resulted in the initial accumulation of 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-delta-enepyranosyluronic acid)-D-galactose (deltaUA GalNAc-0S), and low levels of 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-delta-enepyranosyl uronic acid)-4-O-sulpho-D-galactose (deltaUA GalNAc-4S) in the medium with GalNAc-0S being subsequently utilised during bacterial growth. Metabolic end-products included formate and ethanol but not lactate, indicating that growth was probably carbon-limited. The CS-A contained 30% CS-C, which was also depolymerised resulting in the formation of 2-acetamido-2-deoxy-3-O-(beta-D-gluco-4-delta-enepyranosyluronic acid)-6-O-sulpho-D-galactose (deltaUA GalNAc-6S) in the culture supernate, but this unsaturated disaccharide was apparently not utilised during growth. The results indicate that S. intermedius produced CS-AC depolymerase, which was inducible and extracellular, and sulphatase activity. Experiments with authentic deltaUA GalNAc-4S and deltaUA GalNAc-6S demonstrated that deltaUA GalNAc4S rather than deltaUA GalNAc-6S was the preferred substrate for the sulphatase. Therefore, it is suggested that the CS-AC depolymerase of S. intermedius may play a role in the destruction of CS in host tissues, facilitating bacterial spread, and also in bacterial nutrition by the liberation of nutrients at the site of infection.

Carbohydrate Sequence↗

Intermedilysin is essential for the invasion of hepatoma HepG2 cells by Streptococcus intermedius.

Streptococcus intermedius causes endogenous infections leading to abscesses. This species produces intermedilysin (ILY), a human-specific cytolysin. Because of the significant correlation between higher ILY production levels by S. intermedius and deep-seated abscesses, we constructed ily knockout mutant UNS38 B3 and complementation strain UNS38 B3R1 in order to investigate the role of ILY in deep-seated infections. Strain UNS38 reduced the viability of human liver cell line HepG2 at infection but not of rat liver cell line BRL3A. Isogenic mutant strain UNS38 B3 was not cytotoxic in either cell line. Quantification of S. intermedius revealed that in infected HepG2 cells UNS38 but not UNS38 B3 increased intracellularly concomitantly with increasing cell damage. This difference between UNS38 and UNS38 B3 was not observed with UNS38 B3R1. Invasion and proliferation in BRL3A cells was not observed. Masking UNS38 or UNS38 B3R1 with ILY antibody drastically decreased adherence and invasion of HepG2. Moreover, coating strain UNS38 B3 with ILY partially restored adherence to HepG2 but without subsequent bacterial growth. At 1 day post-infection, many intact UNS38 were detected in the damaged phagosomes of HepG2 with bacterial proliferation observed in the cytoplasm of dead HepG2 after an additional 2 day incubation. These results indicate that surface-bound ILY on S. intermedius is an important factor for invasion of human cells by this bacterium and that secretion of ILY within host cells is essential for subsequent host cell death. These data strongly implicate ILY as an important factor in the pathogenesis of abscesses in vivo by this streptococcus.

Animals↗

Isolation and some properties of fimbriae of oral Streptococcus intermedius.

Streptococcus intermedius 1208-1 carried linear fiber-like fimbriae that extended radially from the cell surface. The fimbriae were isolated by pipetting and sonication and were purified by ammonium sulfate precipitation followed by a column chromatography series. Heat treatment in the presence of sodium dodecyl sulfate resulted in the dissociation into smaller molecules. Rabbit antiserum raised against the purified protein reacted with fimbriae on the surface of bacteria under immunogold staining. Serotype g or g-related strains produced the fimbriae and aggregated in human saliva. The aggregation was inhibited by the anti-fimbriae immunoglobulin Fab fragment or the purified fimbriae.

Antibodies, Bacterial↗

Iliac osteomyelitis and gluteal muscle abscess caused by Streptococcus intermedius.

Streptococcus intermedius, included in the 'milleri group', is a commensal of the mouth and upper respiratory tract but it has often been associated with various pyogenic infections, such as endocarditis, pneumonia, abdominal or cerebral abscess, rarely with osteomyelitis, and exceptionally with muscular abscess. The first observed case of iliac osteomyelitis with gluteal muscle abscess caused by S. intermedius is reported. It is essential to recognise members of the 'milleri group' as possible agents of bone and muscle pyogenic infection because its management requires a timely diagnosis and prolonged antimicrobial treatment to achieve complete clinical and radiological recovery.

Abscess↗

Human transferrin as a source of iron for Streptococcus intermedius.

Streptococcus intermedius is well known to produce severe infections in various areas of the body. In this study, we evaluated the ability of S. intermedius to utilise human transferrin as a source of iron and investigated the mechanism by which iron can be obtained from this plasma protein. Adding either ferrous sulfate or holotransferrin to an iron-deficient culture medium allowed growth of S. intermedius. Cultivation of S. intermedius under an iron-poor condition was associated with the over expression of a 58 kDa cell surface protein. Neither siderophore activity nor reductase activity could be detected. Moreover, cells of S. intermedius did not show transferrin-binding activity or proteolytic activity toward transferrin. It was found that S. intermedius could rapidly decrease the pH of the medium during cell growth, resulting in a release of iron from holotransferrin. When the buffering capacity of the culture medium was significantly increased, the holotransferrin could not support growth of S. intermedius. It is suggested that under certain circumstances, S. intermedius may migrate from its normal niche (oral cavity), reach a particular site and create a localised environment where the pH can be lowered with the subsequent release of iron from transferrin. This would allow bacterial growth and initiation of the infectious process.

Bacterial Proteins↗

Cloning and expression of hyaluronate lyase genes of Streptococcus intermedius and Streptococcus constellatus subsp. constellatus(1).

Hyaluronate lyase (HAase) genes of Streptococcus intermedius and Streptococcus constellatus subsp. constellatus were isolated. In S. constellatus subsp. constellatus, the deduced amino acid sequence of HAase was most similar to that of S. intermedius (68%), whereas the enzyme of S. intermedius was most similar to that of S. pneumoniae (72%). Upstream of the HAase gene on the opposite strands, an open reading frame of a putative glutathione peroxidase started in S. intermedius, and this arrangement was similar to that in S. pneumoniae but unlike that in S. constellatus subsp. constellatus. Cell lysates of Escherichia coli carrying each streptococcal gene showed HAase activity, demonstrating that each cloned gene actually coded for HAase.

Amino Acid Sequence↗

[Cariogenicity of Propionibacterium acnes, Streptococcus intermedius and Streptococcus mutans in germ-free rats].

A comparative study has been made on the cariogenic potentials of Propionibacterium acnes ATCC 11828, Streptococcus intermedius OE-1 and Streptococcus mutans OZ-1. The strains of P. acnes and S. intermedius have chondroitinase activity. As metabolic end-products, the P. acnes strain produces mainly a weak acid, propionic acid, whereas strong lactic acid is the main product of the strains of S. intermedius and S. mutans. Five germ-free rats were used for infection with each bacterial strain. The rats were fed with a 25% sucrose diet for 100 days and then sacrificed. The upper and lower molars were used for the determination of the smooth surface caries score. The right upper and lower jaws were used for the preparation of sections which were employed for the determination of the fissure caries score and microradiography. The left upper and lower jaws were decalcified and used for the preparation of thin sections for staining with Gram's, hematoxilin eosin, Mallory's, alcian blue-PAS, silver and toluidine blue stain. The results obtained were as follows: 1. The recoveries of infected cells in the specimens obtained by swabbing the oral cavities were in the order of 10(4) CFU/mg wet weight regardless of the strains injected. 2. Macroscopically, the P. acnes strain decalcified to a lesser extent some of the enamel surfaces of the lower molars. However, the S. intermedius strain produced mainly the fissure caries of the lower molars. The S. mutans strain destroyed the enamel extensively, resulting in the induction of rampant carious lesions. 3. The caries scores of smooth surface determined by Keyes's method were 19.4 +/- 7.8, 46.8 +/- 5.8 and 118.4 +/- 15.4 in the rats infected with the strains of P. acnes, S. intermedius and S. mutans, respectively. The fissure caries score determined by the modified König's method were 8.5, 22.3 and 36.5 in the sections prepared from the rats infected with the strains of P. acnes, S. intermedius and S. mutans, respectively. 4. Microradiography revealed no translucent area in the enamel and dentine of the P. acnes-infected molars, while in the S. intermedius-infected molars translucent areas were found at the dentino-enamel junction neighboring the bottoms of some fissures. The S. mutans-infected molars showed extensive translucent areas over the enamel and dentine of all of the molars. Even defects of the enamel and dentinal areas were observed in some of the molars. 5. The decalcified tooth sections were stained in a similar manner irrespective of the type of organisms used to infect them.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Functional variation of the antigen I/II surface protein in Streptococcus mutans and Streptococcus intermedius.

Although Streptococcus intermedius and Streptococcus mutans are regarded as members of the commensal microflora of the body, S. intermedius is often associated with deep-seated purulent infections, whereas S. mutans is frequently associated with dental caries. In this study, we investigated the roles of the S. mutans and S. intermedius antigen I/II proteins in adhesion and modulation of cell surface characteristics. By using isogenic mutants, we show that the antigen I/II in S. mutans, but not in S. intermedius, was involved in adhesion to a salivary film under flowing conditions, as well as in binding to rat collagen type I. Binding to human fibronectin was a common function associated with the S. mutans and S. intermedius antigen I/II. Adhesion of S. mutans or S. intermedius to human collagen types I or IV was negligible. Hydrophobicity, as measured by water contact angles, and zeta potentials were unaltered in the S. intermedius mutant. The S. mutans isogenic mutants, on the other hand, exhibited more positive zeta potentials at physiological pH values than did the wild type. The results indicate common and species-specific roles for the antigen I/II in mediating the attachment of S. mutans and S. intermedius to host components and in determining cell surface properties.

Adhesins, Bacterial↗

Fatal Streptococcus MG-intermedius (Streptococcus milleri) meningitis in an adult.

A case of purulent meningitis in an adult caused by Streptococcus MG-intermedius, also known as Streptococcus milleri, is described. The intriguing taxonomical history of this organism and its association with mycoplasma pneumoniae pulmonary infections is reviewed. The incidence of central nervous system infections due to this organism is also discussed.

Cross Reactions↗

Brain abscess due to Streptococcus MG-intermedius (Streptococcus milleri).

Three cases of brain abscesses due to Streptococcus MG-intermedius are reported, and the literature pertaining to this subject is reviewed. The importance of careful and complete identification of these etiological agents of infection is stressed. The clinical presentation, the origin of S. MG-intermedius producing brain abscess, and its relation to hepatic abscesses and endocarditis are discussed.

Adolescent↗