Bacteroides

Haroun N Shah, PHLS Central Public Health Laboratory, London, UK Saheer E Gharbia, University of East London, London, UK
The Bacteroides are a genus of small nonmotile, nonsporing, Gram-negative rods; they are obligate anaerobes and form a major part of the bacterial flora of the human intestinal tract.

Secondary article
Article Contents
. Introduction . History and Current Taxonomic Position . Description of the Genus and Species . Normal Habitat and Physiological Adaptation . Cell Envelope Composition and Antigenic Properties . Disease Association and Pathogenicity

Introduction
Organisms of the genus Bacteroides constitute a major proportion of the bacterial ora of the human intestinal tract. The mechanisms involved in their transition from being a component of the normal ora to becoming a pathogen is one of the challenges of modern microbiology and while many pathogenic determinants have been described, some of these factors are not present in strains that cause infections. It is clear, therefore, that virulence is multifactorial and depends on the physiological status of the host and the genetic diversity of the species.

. Antibiotic Resistance . Conclusion

Description of the Genus and Species

The genus Bacteroides comprises small nonmotile, nonsporing, Gram-negative rods that are moderately pleomorphic. They are obligate anaerobes and grow well at 378C for 2448 h on most blood agar media. Colonies are smooth, shiny, 13 mm in diameter with an entire edge, circular, low-convex and translucent to greyish in appearance. Most strains do not produce haemolysis. All species will grow on a relatively simple medium that contains glucose, haemin, menadione and a few mineral salts (Shah, 1991). They are generally catalase-negative but some strains of B. fragilis, B. thetaiotaomicron, B. vulgatus and B. ovatus possess catalase and superoxide dismutase activities. More recently, the structural gene encoding the superoxide dismutase in B. fragilis was localized to a 581 bp encoding an iron-binding enzyme of 194 amino acids. A good correlation exists between superoxide dismutase activity and oxygen tolerance but no consistent relationship has been found between catalase activity and oxygen tolerance. The mol % G 1 C content of the DNA is within the range 4048 and 16S ribosomal RNA analysis revealed over 93% similarity among species, which places them within the Flexibacter Cytophaga Bacteroides phylum. All species ferment a range of carbohydrates, including glucose, with the production of acid and gas. The major volatile fatty acid products of metabolism are acetic and succinic acids; n-butyric acid is not produced except by B. splanchnicus. The growth of most strains is stimulated by 20% bile but inhibited by sodium deoxycholate. Strains are resistant to penicillin and high concentrations (1 mg per disc) of neomycin and kanamycin. They decarboxylate glutamic acid but nitrate is not reduced and urease is not produced. Most strains hydrolyse aesculin rapidly and produce acid from xylose. Members of the genus are identied by a small number of variable characteristics that include indole production, aesculin hydrolysis and the fermentation of lactose, sucrose, rhamnose, trehalose, mannitol, salicin and arabinose (Table 1).
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History and Current Taxonomic Position

Historically anaerobic Gram-negative, nonsporing rods were assigned to the genus Bacteroides. Initially isolates were mainly from the human alimentary tract but as the ora of dairy, poultry and marine animals were studied, the number of species that conformed to this denition grew immensely. It was evident that heterogeneity existed among species described in various editions of Bergeys Manual but the classication remained unchanged until the 1980s. During this period a large number of techniques were applied and further reinforced the considerable interand intraspecies diversity of this genus. DNA base composition, considered the most reliable criterion for circumscribing the limits of a genus (c. 1012 mol % G 1 C), provided the impetus to restructure the genus, which then comprised species that possessed mol % G 1 C contents of between 28 and 61. Consequently the genus Bacteroides was redened to encompass species that formed a cluster that was phenotypically and genetically akin to the type species, Bacteroides fragilis (Shah and Collins, 1989). Subsequently, two other large groups of species, Prevotella (moderately saccharolytic) and Porphyromonas (nonfermentative), were proposed together with several monospecic genera (see review by Shah, 1991).

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Arabinose Salicin

Bacteroides fragilis
This is the type species of the genus and is the most commonly isolated species of the genus from infections related to the large intestine. It comprises less than 10% of the Bacteroides in normal human faeces but its numbers increase signicantly in infections. Fresh isolates usually form a capsule but are otherwise similar in morphology to other members of the genus. B. fragilis hydrolyses aesculin and produces acid from glucose, lactose, sucrose, maltose and usually xylose, but not from rhamnose, trehalose, mannitol, salicine or arabinose. Indole is not formed and is a useful phenotypic marker for the genus (see Table 1). Virulence is undoubtedly multifactorial and may be attributable to the possession of a polysaccharide capsule or to the action of one or more of the extracellular or membrane-associated enzymes such as proteinases, collagenase, brinolysin, neuraminidase, phosphatase, deoxyribonuclease, hyaluronidase, chondroitin sulfatase and heparinase (Rudek and Haque, 1976). The cell wall of B. fragilis contains a lipopolysaccharide with weak endotoxic activity (Poxton and Edmond, 1995). A species-specic protein component of the outer membrane has been identied and may be used for identication of B. fragilis. The pattern of proteins in the outer membrane shown by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) is also speciesspecic. External to the outer membrane is a thick polysaccharide capsule 1.52 times the thickness of the cell wall. It is composed of a high-molecular weight polysaccharide (7.5 103 kDa), has antiphagocytic properties and also protects the cell from complement-mediated lysis by antibodies against cell wall antigens. The capsular antigen is species-specic. Clinical isolates of B. fragilis are capsulated and may be host-induced (Tzianabos et al., 1994), but are often lost on repeated subculture.
+ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +

Table 1 Physiological profiles of species that currently comprise the genus Bacteroides

Indole production Aesculin hydrolysis

Fermentation of: Glucose Lactose

Sucrose

Rhamnose

Trehalose Mannitol

Xylose

Bacteroides vulgatus
+ + + + + + + + + + +

B. fragilis B. vulgatus B. distasonis B. merdae B. caccae B. ovatus B. thetaiotaomicron B. eggerthii B. variabilis B. uniformis B. stercoris B. splanchnicus

This species was rst isolated from human faeces by Eggerth and Gagnon in 1933. It is the most common Bacteroides species in normal human faeces but is only occasionally implicated in infections. Indole is not produced but charcoalgelatin discs are digested within a few days. Unlike other members of the genus, about 50% of B. vulgatus strains fail to hydrolyse aesculin and others do so only weakly.

+ + + + + + +

Bacteroides distasonis
This species was also described by Eggerth and Gagnon in 1933 in their studies of the faecal ora and named after the Romanian bacteriologist Distaso. Like B. vulgatus, it is a common member of the normal human faecal ora but appears seldom to cause clinical infections. Most strains ferment rhamnose but a signicant minority do not. Indole

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is not produced and charcoalgelatin discs are digested slowly or not at all.

guishes B. eggerthii from other indole-positive species of the genus. Isolates are able to digest charcoalgelatin discs.

Bacteroides merdae
Described by Johnson, Moore and Moore in 1986 in a study of human faecal Bacteroides, this species shares the general characters of the genus. Indole is not produced, charcoalgelatin discs may be digested slowly or not at all, and it is distinguished from other members of the genus by the ability to produce acid from trehalose and salicin, but not from rhamnose or arabinose.

Bacteroides variabilis
This species was described by Distaso in 1912 and is similar to B. eggerthii and to other indole-positive members of the genus, but was included in the species B. thetaiotaomicron in the eighth and ninth editions of Bergeys Manual. It is a common commensal in normal human faeces and isolated from a small proportion of infections in which Bacteroides spp. are implicated. Some strains are inhibited by sodium taurocholate but grow in 20% bile broth. No acid is produced from mannitol or trehalose. Charcoalgelatin discs are usually digested within a few days.

Bacteroides caccae
B. caccae was described in the same study as B. merdae by Johnson, Moore and Moore in 1986, from which it is distinguished by the ability to produce acid from rhamnose but not from salicin.

Bacteroides uniformis
This species was described by Eggerth and Gagnon in 1933. The organisms are normally found in human faeces but rarely implicated in clinical infections. In many studies, strains of B. uniformis have been included with other indole-positive strains as B. thetaiotaomicron. Like B. variabilis, some strains that grow well in 20% broth are inhibited by sodium taurocholate. Charcoalgelatin discs are usually digested within a few days.

Bacteroides ovatus
B. ovatus is one of the less commonly encountered species of the genus. It is not a major component of the normal faecal ora and is isolated only occasionally from clinical specimens. When present in an infection it is usually in large numbers and appears to be a signicant pathogen. It produces acid from a wider range of carbohydrates than other members of the genus. The ability to produce acid from salicin and mannitol is used to identify B. ovatus. Strains produce indole and digest charcoalgelatin discs within a few days.

Bacteroides stercoris
B. stercoris was described by Johnson, Moore and Moore in 1986 in a study of human faecal Bacteroides. Acid is produced from sucrose and rhamnose but not from trehalose, arabinose or salicin and this distinguishes it from other indole-positive species of the Bacteroides.

Bacteroides thetaiotaomicron
B. thetaiotaomicron was named by Distaso in 1912 as a combination of the Greek letters, theta, iota and omicron. It is a common commensal in normal human faeces and the second most common species of the genus isolated from clinical infections, where it appears to have a signicant pathogenic role. Acid is not formed from mannitol by this species and a variable reaction is obtained with the charcoalgelatin disc; some strains digest it readily but others do so only weakly or not at all. In some studies all indole-positive members of the genus that were not B. ovatus were assigned to B. thetaiotaomicron. However, this should be avoided as other indole-positive subgroups or species are known.

Bacteroides splanchnicus
B. splanchnicus has been isolated from normal human faeces and from several infections related to the lower gastrointestinal tract. It shares many characters with other members of the Bacteroides but diers from them in forming signicant levels of n-butyric acid as a metabolic end product. A minority of strains grows well in 20% bile broth but are inhibited by sodium taurocholate. Some strains digest charcoalgelatin discs but others do not. This species shares many chemotaxonomic properties with members of the genus Porphyromonas and the result of 16S rRNA sequence analysis suggests that it is closely related to this lineage.

Bacteroides eggerthii
B. eggerthii was named after the American bacteriologist A. H. Eggerth and described by Holdeman and Moore in 1974 following their studies on the human faecal ora. Acid formation from rhamnose but not sucrose distinENCYCLOPEDIA OF LIFE SCIENCES / & 2001 Nature Publishing Group / www.els.net

Bacteroides

Normal Habitat and Physiological Adaptation

Bacteroides species are numerically one of the most important groups of microorganisms to colonize the human colon and account for about 30% of faecal isolates, with estimates of 109 1010 per gram faeces, but vary with the physiological state of the host (Holdeman et al., 1976). They are metabolically versatile and carry out a wide range of biotransformations that often yield toxic waste products, some of which are potential carcinogens. Their ability to thrive successfully in the colon is due in part to their physiological adaptation to this environment, growth being sustained by the milieu of the colon which contains a complex array of undigested biopolymers. Polysaccharides such as xylans, noncellulosic glucans, pectins, galactomannans, arabinogalactans, mucopolysaccharides, mucin and glycoproteins are actively catabolized by Bacteroides species. Species such as B. fragilis, B. vulgatus, B. ovatus possess several hydrolytic and reductive enzymes such as bglucuronidase, b-glucosidase, arylsulfatase, nitroreductases (McBain and MacFarlane, 1998), which aid in the degradation of these polymers and may be inducible. However, it is now apparent that utilization of polysaccharides does not proceed via breakdown by extracellular polysaccharide-degrading enzymes, but that polysaccharides are initially bound to a putative outer membrane receptor complex and then translocated into the periplasm where the hydrolytic enzymes are located. A cascade of ve genes (susC, D, E, F and G) encode several outer membrane proteins that bind starch and possess amylase and hydrolase activities (DElia and Salyers, 1996). Unlike species from the oral cavity (previously classied as Bacteroides), they can grow in 20% bile and degrade both bile acids and sterols. Most strains of the Bacteroides can deconjugate bile acids and are equally active whether the bile acid is conjugated with glycine or with taurine. Bacteroides thetaiotaomicron converts lithocholic acid to its ethyl ester while Bacteroides fragilis hydrolyses the conjugated metabolites of benzpyrene. In general a wide range of carbohydrates, nitrogenous substrates and various intermediates are catabolized to short-chain fatty acids. They possess enzymes such as fructose-1,6-phosphate aldolase and thus it is assumed that the EmbdenMeyerhofParnas pathway exists (Macy, 1979). However, all species possess enzymes of the hexose monophosphate shunt/pentose phosphate pathway, suggesting that there may be an alternative mechanism for producing phosphoenolpyruvate (PEP) and pyruvate. The former xes carbon dioxide by means of an ADP or GDPdependent PEP carboxykinase in B. fragilis or B. thetaiotaomicron, respectively, to produce oxaloacetate. Pyruvate is reduced to malate by an NAD-dependent malate dehydrogenase. The resulting fumarate is a key intermediate of this pathway as it acts as an electron sink in
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accepting reducing equivalents from various electron donors such as NADH via at least two known electron carriers, protohaem and vitamin K analogues. Both are required by Bacteroides species, protohaem being used for the prosthetic group of cytochromes (generally, b, c or o) while the latter is a precursor for menaquinone biosynthesis. All Bacteroides species possess unsaturated menaquinones to which a polyprenyl side-chain is synthesized in the 3-position of the 1,4-naphthoquinone ring. With the exception of B. splanchnicus and B. distasonis, which produce menaquinones 9 and 10, respectively, all Bacteroides produce signicant levels of both menaquinones 10 and 11. Signicant progress has been made in investigating the mechanisms of polysaccharide utilization among Bacteroides species. The role of outer membrane-binding proteins and hydrolytic enzymes in starch utilization and breakdown of mucopolysaccharides, chondroitin sulfate and heparin has been reported. Evidence from these studies and from physiological analysis of the anaerobic ora of the human digestive tract suggests that Bacteroides species are not capable of utilizing amino acids as nitrogen sources. Therefore, ammonia assimilation is critical for the growth and establishment of these bacteria in the large intestine. Furthermore, the expression of the proteolytic enzymes produced by these species is subject to nitrogen regulation. It was suggested by Yamamoto et al. (1987) that ammonia is assimilated in B. fragilis via the glutamate dehydrogenase (GDH) pathway through the conversion of 2oxoglutarate to glutamate. The 2-oxoglutarate is synthesized via the reductive carboxylation of succinate. Succinate fermentation yields propionate which provides the cell with additional ATP; therefore GDH activity may regulate energy-yielding processes and carbon inux through the regulation of 2-oxoglutarate levels in the cell. Two distinct GDHs are known in both B. fragilis and B. thetaiotaomicron. Dierences in cofactor specicity and mechanism of induction are associated with the two enzymes. The rst enzyme, encoded by the gene gdhA, is a dual cofactor that is NAD(P)H-dependent; it is regulated by the level of ammonia in the culture medium in B. fragilis and its activity is aected by trans-acting genes downstream of gdhA in B. thetaiotaomicron. The second GDH is encoded by a 1355 bp structural gene (gdhB). It is NADHdependent and is induced by the addition of peptides. Sequence homology and comparison of the catalytic active site with other GDHs indicate that both enzymes belong to the family I type hexameric GDH proteins.

Cell Envelope Composition and Antigenic Properties

Cell membranes of Bacteroides species have been shown to contain high levels of sphingolipids or free ceramides. The

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main ester-linked fatty acids are branched-chain anteisoand iso-pentadecanoic acids and the amide-linked fatty acids are all 3-hydroxy fatty acids, also found in lipopolysaccharide (LPS). The ne structure of the cell wall resembles that of other Gram-negative bacteria. All species possess a directly crosslinked peptidoglycan based on meso-diaminopimelic acid. The complex of antigens associated with the outer layer of the cell wall includes the LPS and the outer membrane. The LPS has endotoxic activity, and its polysaccharide is responsible for Oantigenic specicity although it has been shown that puried LPS extracted from dierent strains of B. fragilis had identical immunochemical specicity (see Poxton and Edmond, 1995). Until recently, it was assumed that the LPS of B. fragilis diers from the LPS of aerobic Gramnegative bacilli in that it does not contain either 2-keto-3deoxyoctonate (KDO) or heptoses. The LPS has weak endotoxic activity; it is not lethal for 11-day-old chick embryos when given intravenously and does not induce a local Shwartzman reaction. It gives a positive reaction in the Limulus lysate test, but only at a much higher concentration than the LPS of E. coli. The specicity of O antigens in Bacteroides is determined by the distribution of oligosaccharides, as repeating units in the polysaccharide chain. As with the enterobacteria, O-antigen preparations can be made by boiling suspensions of whole cells. Several workers have attempted to produce a serological typing system for Bacteroides spp. on the basis of agglutination reactions with the O antigens. The genus was later shown to comprise 21 serogroups and 45 serological patterns amongst 98 test strains. Surface-associated components of bacterial cells are now being mapped by the novel technique of matrixassisted laser desorption/ionization time-of-ight mass spectrometry (MALDI-TOF-MS). In this technique, the sample is placed directly on the sample grid and a matrix solution, which facilitates ionization of the molecules, is added and allowed to dry. The sample grid is then placed in a mass spectrometer and a low energy laser is red at the sample. Ions, produced almost instantaneously, travel down a ight tube at a rate that is inversely proportional to their molecular masses and arrive at the detector. The result is revealed as a mass spectral ngerprint of the sample and from their corresponding masses, individual molecules may be deduced. Figure 1 shows the prole of peaks of three isolates of Bacteroides fragilis while a larger number of strains are compared using a dendrogram (Figure 2) to demonstrate the immense strain variation in the surface components of this species.

Figure 1 MALDI-TOF-MS profiles of three strains of Bacteroides fragilis within the mass range 500 1600 Da (a) and 1600 2700 Da (b); strain 12411, a blood culture isolate; 12963, from an abdominal infection; and NCTC 11295, which possesses the enterotoxin gene. A large number of surface components are common to the three isolates but a number of peaks can be seen that distinguish the isolates.

Disease Association and Pathogenicity

Bacterial attributes
Bacteroides species may cause infections anywhere in the body; the most common types are pleuropulmonary, intraabdominal, female genital tract and skin, soft tissue and bone infections. Enzymes such as collagenase, neuraminidase, deoxyribonuclease, heparinase and proteinases are produced by some species and may play a role in pathogenesis. Tissue necrosis and poor blood supply lower the oxidationreduction potential and create favourable conditions for their growth. Therefore, vascular disease, cold, shock, trauma, surgery, foreign bodies, cancer, oedema and gas production by bacteria may signicantly predispose individuals to infection with Bacteroides, as may prior infection with aerobic or facultative bacteria. Infections involving Gram-negative anaerobic bacilli are often characterized by abscess formation and tissue destruction.
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Dissimilarity 12176 12459 12608 13055 NCTC 9343 12366 12190 12411 12174 12294 12293 12185 12653 12410 12075 12700 NCTC 11295 13002 13097 F1 NCTC 8560 12347 12963 12458 12947 12191 12902 12984 13098 12642 12263 12594 12251 12606 0 20 40 60 80 100

Figure 2 A dendrogram showing the relationship of 34 isolates of Bacteroides fragilis based on their MALDI-TOF-MS mass ions. The results show the immense heterogeneity of strains within this species. Interestingly strain F1, from a healthy individual, shows a lower affinity with other strains.

Bacteroides strains isolated from clinical specimens have higher neuraminidase and hyaluronidase activity compared to those isolated from stools. Many species produce phosphatases while the lipopolysaccharides of B. fragilis and B. vulgatus activate the Hageman factor and thereby initiate the intrinsic pathway of coagulation. Strains of B. fragilis vary in their ability to produce an enterotoxin that is implicated as the cause of diarrhoea in calves, piglets, rabbits and more recently humans. Culture
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supernatants from enterotoxigenic B. fragilis strains cause membrane blebbing and reorganization of the F-actin structure of enterocytes. The enterotoxin is a small polypeptide, which is encoded by an open reading frame of 1191 bp encoding a 397-residue holotoxin (Salyers and Shoemaker, 1995). The pre-protein is processed to yield a biologically active toxin of 186 residues with a molecular mass of 20.7 kDa which is secreted into the culture medium. The toxin (known as fragilysin) is a metallopro-

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teinase with a zinc-binding motif. Three distinct subtypes are known among isolates of B. fragilis and share 80% similarity in nucleic acid sequence. Metalloproteinases are important virulence factors among pathogenic bacteria and have been shown to induce metastasis of malignant cells, activate protoxins, cytotoxins and contribute to bacterial invasion and translocation. The presence of this toxin in some of the strains may contribute to the dierence in host response to colonization by B. fragilis.

Host defences against invading Bacteroides spp.

Phagocytic killing by polymorphonuclear leucocytes is mediated by both the oxygen-dependent and oxygenindependent microbicidal systems. Experimental studies indicate that Bacteroides strains are killed under anaerobic and aerobic conditions. Random migration of polymorphonuclear leucocytes is not aected by oxygen tension. Chemotaxis in response to factors generated by bacteria in plasma is markedly depressed under anaerobic conditions. B. fragilis is more resistant to the normal bactericidal activity of serum than are other Bacteroides species. Under anaerobic conditions, B. thetaiotaomicron and B. fragilis are phagocytosed and killed intracellularly by human polymorphonuclear leucocytes only in the presence of normal human serum. There is evidence that surface components interfere with the phagocytosis of capsulated strains. Immunoglobulin and components of the classic and alternative complement pathways participate in chemotaxis, bacteriolysis and opsonization of bacterial strains. Antibody to the capsular polysaccharide of B. fragilis can be induced in animals by infection with encapsulated strains or by implantation of the capsular material itself along with outer membrane components that stimulate an antibody response. Such immunization of animals confers signicant protection against subsequent abscess development from B. fragilis strains. Furthermore, a study of women with acute pelvic inammatory disease demonstrated antibody to the capsular antigen of B. fragilis in women whose infecting ora contained B. fragilis (Paavonen et al., 1981). Immunodiusion analysis of trichloroacetic acid extracts from B. fragilis detected several serotypes.

Antibiotic Resistance
Aminoglycosides such as gentamicin and amikacin are inactive against most anaerobes, as are trimethoprim/ sulfamethoxazole and uoroquinolones. Most penicillins and cephalosporins are less active than penicillin G. Ampicillin, carbenicillin and penicillin V are very eective against 80 to 95% of B. fragilis strains. Over the years

antibiotic resistance has been increasingly reported. Bacteroides spp. have been shown to acquire resistance rapidly to most b-lactam antibiotics (minimal inhibitory concentration usually exceeds 16 mg L 2 1) and to date 93% of resistance strains produce b-lactamases. Both chromosomally mediated and plasmid mediated penicillin resistance have been described, with the production of at least six dierent b-lactamase types. Resistance to chloramphenicol is mediated by either a nitroreductase or an acetyltransferase. Although both mechanisms have been observed among Bacteroides spp., the acetyltransferase activity is transferable among Bacteroides spp. However, the rate of transmission in vivo is apparently low since chloramphenicol resistance is detected in less than 1% of isolates. Clindamycin is one of the most widely used antimicrobial drugs in the treatment of anaerobic infections. Recent studies suggest that resistance to clindamycin in Bacteroides is 520%. Resistance is mediated by an RNA methylase that alters the 50S subunit of ribosomal RNA and modies the site of action of the drug. Resistance is due to the transfer of transposable elements such as Tn4351 and Tn4400. Drugs active against essentially all Gramnegative anaerobes are metronidazole, imipenem, chloramphenicol and combinations of b-lactam drugs plus a blactamase inhibitor. Recent metronidazole resistance in Bacteroides spp. is associated with plasmids that have been shown to transfer to other species via conjugation. This suggests the presence of conjugative elements. Despite the low incidence of resistance to metronidazole, the fact that such resistance is transferable suggests the possibility of increased resistance in the future. Tetracycline resistance in Bacteroides isolates was uncommon, but today all clinical isolates of Bacteroides spp. are resistant to tetracycline due to the spread of conjugative elements throughout Bacteroides spp. Such resistance is mediated by chromosomally expressed ribosomal protection mechanisms. Resistance to macrolides is usually plasmid or transposon mediated and is related to licosamide resistance and results in the dimethylation of the 50S ribosomal subunit. The most frequently detected genes responsible for such activity in B. fragilis, B. ovatus and B. vulgatus are ermF and ermFS. Many Bacteroides strains have cryptic plasmids (36 kb) that are mobilized by conjugative elements. However, clindamycin and erythromycin determinants are harboured on plasmids isolated from B. ovatus. Tetracycline resistance determinants were isolated downstream from these genes. However, the gene products of these tetracycline resistance elements are functional only under aerobic conditions. Conjugative transposons are elements that integrate within the bacterial chromosome but, unlike transposons, do not duplicate the target site. Furthermore, a circular intermediate structure is formed during excision and
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transfer. Bacteroides conjugative transposons usually carry the tetracycline determinant tetQ. Some conjugative transposons carry erythromycin resistance genes that also confer resistance to clindamycin. However, cryptic conjugative elements were detected. Interestingly, strains expressing multiple resistance carry conjugative transposons, which may explain the rapid and widespread occurrence of such resistance among members of the genus Bacteroides. In many cases, conjugative transposons can also mediate the excision and mobilization of co-resident plasmids (Kling et al., 1997). The transfer is stimulated by the exposure of donors to low concentrations of tetracycline prior to conjugation. This was shown to increase the frequency of transfer 1000 10 000-fold. Clindamycin was also shown to stimulate the rate of transfer. Bacteroides conjugative elements have been shown to transfer resistance to Prevotella and Porphyromonas spp. in the presence of tetracycline. Conjugative transposons are extremely stable and, unlike antibiotic resistance plasmids, are maintained in Bacteroides strains in the absence of selection through exposure to antibiotics.

Conclusion
Recognition of members of the genus Bacteroides as a coherent group of species has enabled more focused studies to be undertaken. New species such as Bacteroides aerofaciens are apparent and more will be identied as more in-depth analyses of the bacterial ora of the colon are undertaken. These organisms are indigenous to the intestinal tract and are ideally adapted to this ecosystem but they have the capacity to cause infections at very distant sites. Their remarkable resistance to antibiotics and ability to transform other organisms means that care must be taken in using appropriate treatment. Virulence is multifactorial and specic determinants may be associated with particular genetic variants. Bacteroides fragilis is clinically the most important species but within this and other species a continuum of variants are recognized. Elucidating this diversity in relation to their ecology and pathogenicity may be the key to understanding the nature of pathogenicity of these microorganisms.

Holdeman LV, Good IJ and Moore WEC (1976) Human fecal ora: variation in bacterial composition within individuals and a possible eect of emotional stress. Applied and Environmental Microbiology 31: 359375. Kling JJ, Wright RL, Moncrief JS and Wilkins TD (1997) Cloning and characterization of the gene for the metalloprotease enterotoxin of Bacteroides fragilis. FEMS Microbiology Letters 146: 279284. Macy JM (1979) The biology of gastrointestinal Bacteroides. Annual Review of Microbiology 33: 561594. McBain AJ and MacFarlane GT (1998) Ecological and physiological studies on large intestinal bacteria in relation to production of hydrolytic and reductive enzymes involved in formation of genotoxic metabolites. Journal of Medical Microbiology 47: 407416. Paavonen J, Valtonen VV, Kasper DL et al. (1981) Serological evidence for the role of Bacteroides fragilis and Enterobacteriaceae in the pathogenesis of acute pelvic inammatory disease. Lancet 1(8215): 293295. Poxton IR and Edmond DM (1995) Biological activity of Bacteroides lipopolysaccharide reappraisal. Clinical Infectious Diseases 20 (suppl. 2): S149153. Rudek W and Haque RU (1976) Extracellular enzymes of the genus Bacteroides. Journal of Clinical Microbiology 4: 458460. Salyers AA and Shoemaker NB (1995) Conjugative transposons: the force behind the spread of antibiotic resistance genes. Anaerobe 1: 143150. Shah HN (1991) The genus Bacteroides and related taxa. In: Balows A, Tru per HG, Dworkin M, Harder W and Schleifer KH (eds) The Prokaryotes, 2nd edn, pp. 35933607. New York: Springer-Verlag. Shah HN and Collins MD (1989) Proposal to restrict the genus Bacteroides (Castellani and Chalmers) to Bacteroides fragilis and closely related species. International Journal of Systematic Bacteriology 39: 8597. Tzianabos AO, Onderdonk AB, Smith RS and Kasper DL (1994) Structurefunction relationships for polysaccharide induced intraabdominal abscesses. Infection and Immunity 62: 35903593. Yamamoto I, Saito H and Ishimoto M (1987) Regulation of synthesis and reversible inactivation in vivo of dual coenzyme-specic glutamate dehydrogenase in Bacteroides fragilis. Journal of General Microbiology 133: 27732780.

Further Reading
Duerden BI and Drasar BS (1991) Anaerobes in Human Disease. London: Edward Arnold. Finegold SM and Lance George W (1989) Anaerobic Infections in Humans. New York: Academic Press. Gharbia SE, Williams JC, Andrews DMA and Shah HN (1995) Genomic clusters and codon usage in relation to gene expression in Gram-negative anaerobes. Anaerobe 1: 239262. Salyers AA (1984) Bacteroides of the human lower intestinal tract. Annual Review of Microbiology 38: 293313. Shah HN, Gharbia SE and Duerden BI (1998) The genera Porphyromonas, Prevotella and Bacteroides. In: Topley and Wilsons Microbiology and Microbial Infections, vol. 2, 9th edn, chap. 58, pp. 1305 1330. London: Arnold.

References
DElia JN and Salyers AA (1996) Contribution of a neopullulanase, a pullulanase, and an alpha-glucosidase to growth of Bacteroides thetaiotaomicron on starch. Journal of Bacteriology 178: 71737179.

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