lacking the enzyme catalase, Oral streptococci are clustered into four groups (see Table 3.3) and will now be described. ‘Matans group (mutans streptococci) ‘There is great interest in the mutans streptococci because of their potential role in the aetiology of dental caries. Streptococcus mutans was originally iso- lated from carious human teeth by Clarke in 1924 and shortly afterwards, was recovered from a case of infec- tive endocarditis (growth of bacteria on damaged heart valves; Chapter 11). Lite attention was paid to this species until the 1960s when it was demonstrated that caries could be experimentally-induced and transmit- ted in animals artificially infected with strains resem- bling S. mutans. The name of this species derives from the fact that cells can lose their coccal morphology and often appear as short rods or as coccobacilli. Nine serotypes have been recognised (a-h, and k), based on serological specificity of carbohydrate antigens located. in the cell wall, although some serotypes are found only in animals. Subsequent work showed that suffi- cient differences existed between clusters of these serotypes to warrant subdivision into seven distinct species (Table 3.3); these species are described col- lectively as mutans streptococci. Mutans streptococci are recovered almost exclusively from hard, non- shedding surfaces in the mouth, such as teeth or den- tures, and can act as opportunistic pathogens, being isolated from cases of infective endocarditis (Chapter 11). Mutans streptococci are regularly isolated from dental plaque at carious sites, but their prevalence is ow on sound enamel, The specific epithet, S. mutans, is now limited to Jhuman isolates previously belonging to serotypes ¢,¢, fand k. This is the most commonly isolated species of ‘mutans streptococci, and epidemiological studies have implicated S. mutans as one of the main causative organisms in the aetiology of enamel and root surface caries (see Chapter 6). The next most commonly iso- lated species of the mutans streptococci group is S. sobrinus (previously, S. mutans serotypes d and g) and is also associated with human dental caries. Less is mown about the role of S. sobrinus in disease because some studies do not attempt to distinguish between these species, and some commonly used selective agar media for the isolation of mutans streptococci contain 8. The Resident Oral Microbiota = 35 bacitracin, which can be inhibitory to the growth of both S. sobrinus and S. criceti (formerly 5. cricets, and previously termed S. mutans serotype a). Some people harbour more than one species of mutans streptococci in their mouth. The antigenic structure of mutans streptococci has been studied in detail to establish serological typing schemes and for the development of a prospective caries vaccine (see Chapter 6). Mutans streptococci possess cell wall carbohydrate antigens, lipoteichoic acid, lipoproteins and cell wall or cell wall-associated proteins. Antigen VI (also termed antigen B, SpaP or Pac) has generated considerable interest because it is (a) a major adhesin involved in the initial adherence of S. mutans to the tooth surface (See Chapter 5) and (b) a possible component of a subunit caries vaccine (ee Chapter 6). Some strains of S. mutans carry a collagen-binding gene, and these strains have been isolated from patients with cerebral microbleeds. Mutans streptococci make extracellular soluble and {insoluble polysaccharides (glucan, mutan and fructan) from sucrose that are associated with dental plaque ‘maturation (see Chapters 4 and 5) and cariogenicity ee Chapter 6). The glucans and fructans are pro- duced by glucosyltransferases and. fructosyltrans- ferases, respectively: Mutan isa highly insoluble glucan. that is only produced by mutans streptococci, whereas the fructan is unusual in having an inulin-like struc- ture. These polymers contribute to the characteristic colonial morphology of mutans streptococci when growing on sucrose-containing agar plates (Fig. 3. B), Mutans streptococci can also synthesise intracel- lular polysaccharides when there is excess sugar, and. these can act as carbohydrate reserves and be con- verted to acid during periods when dietary carbohy- ddrates are in limited supply. Mutans streptococci can scavenge dietary sugars very efficiently and rapidly convert them to acidic fermentation products (mainly lactate), Significantly, mutans streptococci aré able to grow and survive under the acidic conditions they generate by the induction of specific molecular stress responses (see Chapter 4 and Fig. 2.4). Mutans strep- tococci can communicate with other mutans strepto- cocci by the release of diffusible signalling molecules that can induce genetic competence (an ability to take uup extracellular DNA) and acid tolerance in neigh- bouring cells36 Marsh and Martin's Oral Microbiology elctron microscopy (SEM). (B) The colony morphology of Streptococcus mutans growing on sucrose-contaning agar. (C) Gram stain of Actinomyces israeli. (0) Gram stan of Eubacterium yuri. Grand kage were Kindly SEM was kindly provide by Wendy Rowe, Cra University Salivarius group This group comprises S. salivarius and S. vestibulars Strains of S. salivarius are commonly isolated from ‘most areas of the mouth, although they preferentially colonise mucosal surfaces, especially the tongue Streptococcus salivarius produces large quantities of an ‘unusual extracellular fructan (polymer of fructose with a levan structure) from sucrose (see Chapter 4), as well as a levanase that can degrade this type of fructan. This levan gives rise to characteristically large mucoid colonies when S. salivarius is. grown on sucrose-containing agar. Streptococcus salivarius also produces extracellular soluble and insoluble glucans from sucrose; some strains have urease activity. Strep tococcus salivariu is isolated only rarely from diseased ded by Owain Dayo Thoras, Carat and Vale UHB, sites and is not considered a significant opportunistic thogen. Streptococcus vestibularis is isolated mainly from the vestibular mucosa of the human mouth. These bacte- ria do not produce extracellular polysaccharides from sucrose, but do produce a urease (which can generate ammonia and hence raise the local pH) and hydrogen peroxide (which can contribute to the sialoperoxidas system [see Chapter 2], and inhibit the growth of competing bacteria) Anginosus group Representative species of this group, Streptococcus constellatus (subspecies constellatus, subspecies vibor gensis and subspecies pharyngis), S. intermedius andS. anginosus (subspecies anginosus and. subspecies whileyi), are readily isolated from dental plaque and from mucosal surfaces and are important causes of serious, purulent disease in humans, including maxil- lofacial infections. Members of the anginosus group are commonly found in abscesses of internal organs, especially of the brain and liver, and have also been recovered from cases of appendicitis, peritonitis, men- ingitis and endocarditis. Streptococcus intermedius is isolated mainly from liver and brain abscesses, whereas S. anginosus and S. constellatus are derived from puru- lent infections from a wider range of sites. Streptococ- cusintermedius can produce acytotoxin, intermedilysin, which may also interfere with neutrophil function and. enable the cell to evade the host defences during, abscess formation. This group does not make extracel- Tular polysaccharides from sucrose Mitis group The application of molecular phylogenetic techniques (involving the determination of 165 rRNA gene sequences) has resolved many of the previous anoma- lies in the classification of this group, resulting in the identification of new species. ‘Streptococcus sanguinis and S. gordonii are early colo- nisers of the tooth surface, and both produce extracel- lular soluble and insoluble glucans (see Chapter 4) from sucrose that contribute to biofilm formation Both species can generate ammonia from arginine. S. sanguinis produces a protease that can cleave slgA (IgA protease), whereas S. gordonii can bind salivary -amylase enabling these organisms to break down starch. Amylase-binding may also mask bacterial anti- gens and allow these bacteria to avoid recognition by the host defences (host mimicry). Both species are composed of several biotypes. ‘wo of the most common streptococcal species in the mouth are S. mitis and S, oralis. Strains of S. oralis produce neuraminidase (an enzyme that removes sialic acid from oligosaccharide side chains of salivary mucins) and an IgA protease, but cannot bind d-amylase (Fig. 3.5, A). Streptococcus mitis is subdivided into two biotypes, and these show different distribution patterns in the mouth. Strains of these two species are able to take up extracellular DNA (Le., they are genetically competent), and this process is facilitated in biofilms like dental plaque where bacteria are in close 2 Meredoaincton a7 | Proximity to one another. Consequently, it is perhaps hot surprising that there is considerable genetic and phenotypic heterogeneity when the properties of lange numbers of S. mitis and S. oralis strains are compared. Some, but not all, strains of these two species are able to produce extracellular glucan from sucrose. Other members of this group include S. parasan- uinis (formerly S. parasanguis) that has been isolated from clinical specimens (throat, blood, urine). Strains can hydrolyse arginine but not urea, and can bind salivary g-amylase, but cannot produce extracellular polysaccharides from sucrose. Streptococcus cristatus is characterised by the presence of tufts of fibrils on their cell surface. Newer species have been described including 5, oligofermentans, S. sinensis, S, australis, S infantis, S. peroris, S. tigurinus and S. dentisani. The significance of some of these species to the ecology of the mouth has yet to be determined, but species such as S. tigurinus and S. sinensis can act as opportunistic pathogens, having been isolated from cases of infective endocarditis, Members of the mitis group are opportunistic pathogens, particularly in infective endocarditis (see Chapter 11). Streptococcus pneumoniae can be isolated from the nasopharynx and is a significant opportun- istic pathogen, which can acquire and transfer antibi- otic resistance genes amongst other members of the itis group. OTHER GRAM-POSITIVE COCCI Surains that were originally described as being nutritionally variant. streptococci (NVS) have been isolated from the mouth when appropriate isolation ‘media are used. These have been reclassified as Granu- licatelta adiacens (previously S. adiacens and Abiotrophia adiacens) and Abiotrophia defectiva (previously S. defeci- vyus). Granulicatella adiacens is common in the mouth and is an early coloniser of the tooth surface, although it is overlooked in most studies because of the need for isolation media to be supplemented with growth factors such as cysteine or pyridoxal. These bacteria often exhibit satellitism, seen as an enhanced growth pattern around colonies of certain other bacteria that produce these cofactors. Other Gram-positive cocci include Gemella species (G. haemolysans and G. morbil- lorum), although cells sometimes appear Gram-negative on staining,38 Marsh and Martin's Oral Mirebioloay Anaerobic Gram-positive cocci are_ commonly recovered from dental biofilms, especially fom carious dentine, infected pulp chambers and root canals (see Chapter 6), advanced forms of periodontal disease Gee Chapter 6) and from dental abscesses (see Chapter 7), These bacteria are also recovered from deep-seated abscesses elsewhere in the body and are ‘usually isolated in mixed culture (polymicrobialinfec- tions). The taxonomy of this group of organisms has been clarified. Originally, strains were placed in the genus, Peplostreptococcus, and representative species Included P micros, magnus and Panaerobius. However, P micros and P magnus have been moved to new genera and are now called Parvimonas micra and Finegoldia ‘magna, respectively, whereas oral strains of P anaero- bius are now designated Peptstrepracoccus stomats Enterococci have been recovered in low numbers from several oral sites wien appropriate selective media have been used; the most frequently isolated species is Enterococcus faecalis, Enterococci can be iso- lated from the mouths of immuno-compromised and edically-compromised patients, and have been iso- lated from periodontal pockets that fil to respond to therapy and from infected root canals. Lancefield group A streptococet (S. pyogenes) are not usually i0- lated from the mouth of healthy individuals, although, they can ofien be cultured from the saliva of people suffering from streptococcal sore throats and may be associated with a particularly acute form of gingivitis (ee Chapter 6). Staphylococci and micrococci are also not com- monly isolated in lange numbers from the oral cavity, although the former are found in denture plaque, as ‘well as in immunocompromised patients and indi- viduals suffering from a variety of oral infections (see Chapters 7 and 11). These bacteria are not usually considered to be members of the resident oral micto- biota, but they may be present transiently, and they have been isolated from some sites with root surface caries and from some periodontal pockets that fail 0 respond to conventional therapy. This isin sharp con- trast (0 other surfaces of the human body in close proximity to the mouth, such as the skin surface and the mucous membranes of the nose, where they are among the predominant components of the microbi- ota, This finding emphasises the major differences that ‘must exist in the ecology of these particular habitats Microorganisms fom the mucosal surfaces of the skin and nose must be passed consistently into the mouth and yet these organisms are normally "unable to colonise or compete against the resident oral rierobiota GRAM-POSITIVE RODS AND FILAMENTS ACTINOMYCES ‘Actinomyces species form a major portion of the mnicto- biota of dental plaque, particulary at approximal sites and the gingival crevice, These bacteria have been associated with root surface caries, and their mumbers increase during gingivitis (See Chapter 6). Cells of ‘Actinomyces species appear as short rods but are often pleomorphic in shape; some cells show a true branch- ing morphology, whereas these of A. israeli can some- times appear to be filamentous (Fig, 3.5, C). Some species (particularly A. naeslundi) are heavily fimbri- ated (cell surface structures involved in attachment), whist others have relatively smooth surfaces. Some newly described species have been identified in a variety of clinical specimens (A. radingae, A. neui, A Johnson, A. europacus, A. graevenitei, A, funkei, A. den- talis and A. turicensis), including infective endocarditis and abscesses, but the source and habitat of these species is not yet fully understood. Actinomyces radici- dentis has been isolated from endodontic infections. The most common Gram-positive bacillus in plaque is Actinomyces naeslundii, which was originally subdivided into two genospecies (4. naeslundit geno- species 1 and genospecies 2). Actinomyces naeslundi _genospecies 2 is now classified as A, oris. Some strains of A. nacslundit produce an extracellular slime and a fructan from stctose witha levan-like structure as well as enzymes that can hydrolyse fructans, Some strains also produce urease (this enzyme may have a role in ‘modulating pH in plaque) and neuraminidase (can ‘modify receptors in the enamel acquired pellicle). Two types of fimbriae can be found on the surface of cells of A, naeshindi. Each type serves a specific function, and are implicated either in cell-to-cell contact (coag. sgregation) or in cell-to-surface interaction (see Chapter 5). Actinomyces viscosus is a closely related species found in animals Actinomyces israeli cam act as an opportunistic pathogen causing a chronic inflammatory condition