INTERNATIONALJOURNAL OF SYSTEMATICBACTERIOLOGY, Oct. 1990, p. 426-433 Vol. 40, No. 4 0020-7713/90/040426-08$02.00/0 Copyright 0 1990, International Union of Microbiological Societies

Transfer of Kingella indologenes (Snell and Lapage 1976) to the Genus Suttonella gen. nov. as Suttonella indologenes comb. nov. ; Transfer of Bacteroides nodosus (Beveridge 1941) to the Genus Dichelobacter gen. nov. as comb. nov. ; and Assignment of the Genera , Dichelobacter, and Suttonella to fam. nov. in the Gamma Division of on the Basis of 16s rRNA Sequence Comparisons

FLOYD E. DEWHIRST,l* BRUCE J. PASTER,, SHARON LA FONTAINE,3 AND JULIAN I. ROOD3 Departments of Pharmacology' and Microbiology,2 Forsyth Dental Center, Boston, Massachusetts 021 15, and Department of Microbiology, Monash University, Clayton 31 68, Australia3

The 16s rRNA sequences of Kingella indologenes, Cardiobacterium horninis, and Bacteroides nodosus were determined by direct RNA sequencing, using a modified Sanger method. Sequence comparisons indicated that these three species represent a novel family in the gamma division of Proteobacteria. On the basis of these data, K. indologenes and B. nodosus cannot retain their current generic status as they are not closely related to other members of their assigned genera. Therefore, we propose transfer of K, indologenes to the new genus Suttonella as Suttonella indologenes and transfer of B. nodosus to the new genus Dichelobacter as Dichelobacter nodosus and assign the genera Cardiobacterium, Suttonella, and Dichelobacter to a new family, Cardiobacteriaceae, in the gamma division of Proteobacteria.

Comparison of 16s rRNA sequences has proven to be goats, and cattle (33). Infected hooves are the only known extremely useful for determining phylogenetic relationships habitat of Bacteroides nodosus. Previous reports have also among eucaryotic and procaryotic organisms (46). Previ- indicated that Bacteroides nodosus is not closely related to ously, we sequenced the 16s rRNAs of a number of micro- other species of the genus Bacteroides and is generically organisms, including several species in the family Neisseri- misnamed (16, 19, 31, 37). Below we use brackets in refer- aceae, to determine the phylogeny of the beta division of ring to [Kingella] indologenes and [Bacteroides] nodosus to Proteobacteria (10). Sequence comparisons indicated that indicate that they do not belong in these genera. two of the microorganisms which we examined, Kingella indologenes and , are not closely MATERIALS AND METHODS related to the , but rather are related to one Bacterial strains and culture conditions. The sources of the another as members of a novel family in the gamma division strains which we examined in this study are shown in Table of Proteobacteria. A subsequent comparison of a partial 1. Cardiobacterium hominis and [Kingella] indologenes sequence of the cloned 16s rRNA gene of Bacteroides were cultured aerobically at 37°C in commercially available nodosus (19) with the sequence of Kingella indologenes and Todd-Hewitt broth (BBL Microbiology Systems, Cockeys- Cardiobacterium hominis indicated that Bacteroides no- ville, Md.). [Bacteroides] nodosus was cultured on TAS dosus also belongs to this novel family. We determined (Trypticase-arginine-serine) agar at 37°C in an 80% N,-10% virtually the complete rRNA sequence of Bacteroides no- CO,-lO% H, atmosphere as described by Skerman (32). dosus in order to have a comparable number of bases for Isolation and purification of rRNA. rRNA was isolated and sequence comparisons with other species in our database. partially purified by using a modification of the procedure of The sequences of Cardiobacterium hominis, Kingella in- Pace et al. (24), as previously described (25). dologenes, and Bacteroides nodosus were compared with 16s rRNA sequencing. rRNA was sequenced by using a the sequences of nine beta division proteobacteria and nine modified Sanger dideoxy chain termination technique in gamma division proteobacteria in order to determine the which primers complementary to conserved regions were phylogenetic relationships among these . elongated by using reverse transcriptase (20). The details of Cardiobacterium hominis is an occasional resident of our protocol have been described previously (10, 25). human respiratory tracts and has been recovered from blood Data analysis. A program set for data entry, editing, samples of humans with endocarditis (18, 34, 45, 48). Kin- sequence alignment, secondary-structure comparison, simi- gella indologenes has been recovered from human eye larity matrix generation, and dendrogram construction for infections and from the blood of a patient with endocarditis 16s rRNA data was written in Microsoft QuickBASIC for (15, 39, 42). It has been reported previously that Kingella use on IBM PC-AT and compatible computers (25). RNA indologenes is not closely related to other Kingella species sequences were entered and aligned as previously described (10, 28) and thus is generically misnamed. Bacteroides (25). Our sequence database contains approximately 200 nodosus is the essential causative agent of footrot in sheep, sequences, including sequences determined in our labora- tory, previously published sequences, and unpublished se- quences provided to us by other scientists. Nine beta and * Corresponding author. nine gamma division proteobacteria were chosen for com-

426 VOL.40, 1990 CARDIOBACTERIACEAE FAM. NOV. 427

TABLE 1. Sources and accession numbers of the strains studied tree was calculated by using a modified unweighted pair group method in which differing branch lengths reflected Accession Refer- Organism StrainU no.‘ ence“ differing numbers of base changes relative to the other species in the tree. The root of the tree was established by Sequenced organisms performing several analyses, using multiple alpha or beta Cardiobacterium hominis ATCC 16826T M35014 division proteobacteria or more distantly related eubacteria [Kingella]indologenes ATCC 25869T M35015 as outgroups. Cardiobacterium hominis, [Kingella] indolo- [Bacteroides]nodosus 198A M35016 Reference organisms: gamma genes, and [Bacteroides] nodosus formed a cluster with an division average level of similarity of 93%; we refer to this cluster rrnB cistron 501695 4 below as the Cardiobacteriaceae cluster. This cluster Monteil 501874 6 branched deeply within the gamma division of the Proteo- Actinobacillus lignieresii ATCC 19393T M35017 A bacteria. The position of the tree root was somewhat sensi- NCTC 10322T M35018 A tive to selection of the outgroup. The Cardiobacteriaceae ATCC 33391T M35019 A cluster occasionally branched as a deep member of the beta Ruminobacter amylophilus DSM 1361T NA 22 division of the Proteohacteria. An analysis of positions Oceanospirillum linum ATCC 11336T M22365 B which normally differentiate the beta and gamma divisions of ATCC 25330 M34133 B Chromatium vinosum M. Madigan M26629 B the Proteobacteria indicated that the organisms included in Reference organisms: beta the Cardiobacteriaceae cluster possessed 8 gamma division division signatures and 4 beta division signatures, whereas the ATCC 23834T M22512 10 slightly less deeply branching organism Chromatium vino- Neisseria denitrijkans ATCC 14686T M35020 A sum and other gamma division organisms possessed 12 of 12 Kingella denitrijicnns ATCC 33394T M22516 10 gamma division signatures and the 9 beta division species NCTC 8375T X07714 27 contained 12 of 12 beta division signatures. Thus, the results ATCC 23330T M22517 10 of the signature analysis supported placement of the Cardio- Spirillum volutans ATCC 19554T M34131 B bacteriaceae cluster as a deep branch of the gamma division Alcaligenes faecalis ATCC 8750T M22508 10 Rhodocyclus pupura M. Madigan M34132 B of the Proteobacteria. [Pseudomonas]cepacia ATCC 25416T M22518 10 DISCUSSION a ATCC, American Type Culture Collection, Rockville, Md.; DSM, Deut- sche Sammlung von Mikroorganismen, Braunschweig, Federal Republic of Germany; NCTC, National Collection of Type Cultures, London, United Until recently, the phylogenetic position of Cardiobacte- Kingdom. rium horninis relative to other genera was unknown, as Sequences are available for electronic retrieval from GenBank under the demonstrated by the placement of this organism in “other accession numbers given. These sequences should also be available from the genera of facultatively anaerobic gram-negative rods” in European and Japanese collections. NA, Not available from GenBank. A, Unpublished sequences deposited in GenBank by F. E. Dewhirst and Bergey ’s Manual of Systematic Bacteriology (45). There was B. J. Paster; B, unpublished sequences deposited in GenBank by C. Woese. suspicion that Cardiobacterium hominis was related to the family Neisseriaceae, although no DNA-DNA hybridization (40) or genetic transformation (41) was observed. While our parison with the three sequenced organisms. The reference work was in progress, rRNA cistron analysis placed Cardio- strains, their GenBank accession numbers, and literature bacterium hominis between rRNA superfamilies I and I1 and references are shown in Table 1. Phylogenetic trees were rRNA superfamily 111 (9, 28), which is equivalent to our constructed by using the modified unweighted pair group placement of this organism at the branching point between method of Li (21). the beta and gamma divisions of the Proteobacteria. Pheno- typic traits of Cardiobacterium hominis are shown in Table 3. RESULTS The finding that [Kingella] indologenes is not closely By using seven primers, 1,463,1,466and 1,529 bases were related to other Kingella species but rather is a deeply determined for [Kingella] indologenes, Cardiobncterium branching member of the gamma division of the Proteobac- hominis, and [Bacteroides] nodosus, respectively. The se- teria which is related to Cardiobacterium hominis (Fig. 1) is quences were aligned with and numbered relative to the consistent with the results of rRNA cistron analysis (28, 29). Escherichia coli sequence (4) (Fig. 1). The sequence given The admonition by Snell that “the genus Kingella is a for [Bacteroides] nodosus is a consensus sequence based grouping of convenience and further taxonomic study is upon the previously published partial sequence from cloned needed to test its robustness” was well founded (35). [Kin- DNA (19) and the virtually complete sequence derived from gella] indologenes is clearly not closely related to Kingella Sanger sequencing of the 16s rRNA. The sequence shown in kingae, the type species of the genus, or to Kingella deni- Fig. 1 is in agreement with the previously published partial trijicans, which are members of the family Neisseriaceae in sequence (19); however, the new sequence provides infor- the beta division of Proteobacteria. While Kingella kingae mation for positions 2 to 608 and 1009 to 1017. The positions and Kingella denitrijicans are relatively closely related, they where the three organisms have unique or highly unusual fall into different clusters within the family Neisseriaceae single base signatures are indicated on Fig. 1, as are the base and may eventually have to be placed in separate genera (10, positions which usually differentiate between the beta and 28; unpublished data). [Kingella] indologenes is phenotypi- gamma divisions of Proteobacteria. Sequence similarity cally very similar to Cardiobacterium hominis (Table 3). matrices for [Kingella] indologenes, Cardiobacterium hom- However, the guanine-plus-cytosine (G+C) contents of the inis, [Bacteroides] nodosus, and reference beta and gamma DNAs of these species differ by 11 mol%, and their 16s division proteobacteria are shown in Table 2. rRNA sequences differ by 896, which suggests that they Figure 2 is a dendrogram which shows the phylogenetic should be placed in sister genera rather than in the same relationships of the organisms included in this study. The genus. Therefore, we propose that [Kingella] indologenes a r Ch ....A~GAGWUgallllCUGGCUCAGAWGMCGCUGGCGGCAUGC~MCACAUGCMWCtMCGG~CGA--UGGAGCUUGCUCCA--GGCWCGAGUGGCGMCGGWGAGUAA Ch Si WWACUWACAWUIGAI*ICUGGCUCAGAWtMCGCUGGCGGCAUGCUUMCACAUGCMWCGMCGAGGGA----AGCAGCWGCUGCn----CACCUffiUGGCGGACGGWGAWAA Si Dn ...MCUWACAWU1CAurCUGGCUCAGAWWACGCUGGCGGCAUGCUu~CACAUGCMWCGMCGGG~A---UWAGCUUGCUAUG---UMCCUAGUGGCGGACGGGUGAGUAA Dn Ec MAWWACACUUUWUCAUGGCUCA~WWACGCUGGCGGCAGGCCUMCACAUGCMCUCGMCGGUMCAGWAGMGCWGCWCUGCU~CGAGUGGCGGACGGWGACUAA Ec 10 20 30 40 50 60 70 80 90 100 110 120

Ch CGCAUGGWAUCUGCCUUlUGCUGGGGGAUMCGUAGGGAMCUUACGCUMUACCGCAUMCACCUMGGGU~GCGGGGGACCGAA-AGCCUCGCGGCAAGAGAUGAGCCCAUGW Ch Si CWAUAGWAUCUACCUuCGGWGGGgGAUAACGUAUGCAMCGUACCCUAAUACCACAUMCACWACGAGUC~GCGGGGGAUCWCGGACCUCGCGCCCWAGAUGAGCCUAUAW Si Dn UAUAUAGWAUCUGCCWAUGGUGGGGWCAACGUAUGGAMCGUACGCUMUACCGCAUMCAWWAGAAU~GCGGGGGCUCGAAAGACCUCGCCCCWMGAUGAGCCUAUAUC Dn Ec UWCUGGWMCUGCCUCAUGGAGGGGGAUMCUACUGGAMCGGUAGCUMUACCGCAUMCWCGCMGACCAMCAGGGGGACCWCGGGCCUCWGCCAUCGCAUGUGCCCAGAUG Ec 130 140 150 160 170 180 190 200 210 220 230 240

Ch GCAWAGCUAGUJGWGGGWMAgGCCUACCMGGCCACCAUCCAUAGCUGGUCUCACAGCAUCAUCAGC~CACUGGCACUCAGACACGGCCCaCAtUCC~CGGGAGGCAGCAWGGCh Si GCAWACCUACUlCWRGtAMGGCCUACCMGGC~CCAUCCAUAGCUG~~~GCAUCAUCAGCCACAUCGGCACUGACACACGGCCC~CUCcUACGGCAGGCAGCAWGGSi Dn CCAWAGEUACUlGWGGGCMCAGC~UACCMGGCCACCAUCCWAGCUG~GACAtMUCAUCAGCCACAUCGGCACUCACACACGGCCC~CUCCUACGGGAGGCAGCAWGGOn Ec GCAWACEUAWAGWGGGWMCGGCUCACCUAGGCCACCAUCCCUAGCUGWCUWCAGCAUCACCAGCCACACUGWACUWCACACGWCCACACUCCUACGGGAGGCAGCAWGG Ec 250 260 270 280 290 300 310 320 330 340 350 360

Ch GWAUAWGGaCnAUGGGGGGMCCCUCAUCCAGCMUGCCGCWWGUGMCMGGCCWCGGGUJWAMGCACUUCAWAGGGAGGAMGWGCWA~AAUACCUGCGCM-W Ch Si GWAUAUIGGnCnAUGGGGGCMCCCUCAUCCAGCMUGCCGCGUWWtMGMGGCCUJffiGGUJWAMGCA~CWUAGUGMGAMG~AUGGWAAUACCUGUWAU-W Si Dn GWAUaWlCCACnAUGGGGGGMCCCUCAUCCAGCMUGCCGCGUWWGAAGAAGGCCUJCGGMWAMGCACAWA~GAAGAACGWGCAUG~AAUACCCAUGCAA-UlDfl Ec GWAUAWGCACMUGGGCGCAAGCCUCAUGCAGCCAUGCCGCWWAUGAAGAAGGCCWCGGGWWAAAGUACUCAGCGGGGAGGAAGGGAWMA~AAUACCUUUGCUCAUl Ec 370 380 390 coo 410 420 430 440 450 460 470 480 r Ch GACGUJACCUACAWAGMGCACCGGCUMCUCCGUGCCAGCAGCCGCGGUAAUACGGAGGGUGCCAGCGWAWCGGMWACUGGGCGUAMGCGCACGCAGGCG~GCCCAAGUCA Ch Si CACAWAGCUaMWAWAGCACCGGCUaACuCCWGCCAGCAGCCGCGGUMUACGGAGG~GCnAGCCAWcGGAAUGACUGGGCGUAMGCGCACGCAGGCGGWAUWMWCA Si Dn CAUWAGCUaAGWAAMGCACCGGCUnACUCCWGCCAGCAGCCGCGWaAUACGCAGGWGCMGC~AWCGGMUCACUGGGCGUAMGCGCACGCAGWG~AUAAGUCADn Ec GACGUJACCCGCACMWAGCACCGGCUAACUCC~GCCAGCAGCCGCGWMUACGGAGGWGCMGCGWAAUCGGAAWACUGGGC~AAAGCGCACGCAGGCGGWUCUlAAGUCA Ec 490 500 510 520 530 540 550 560 570 580 590 600 8 Ch CAUWWMGCCCCGGGCMCCUGGCMCUGCA~~CUGGGCGAC~ACA~UWMGAGWMGCGWAUCCAWWAGCAWGAAAUGCWACAUAWGGAAGGAACACCCh Si CAUWWMGCCCCGGGCUJMCCUGGGnAWGCAU~CUGGWMtUACAWAUWCACAGWAGGCGtMUCCAWWAGCAWGAMUGCWACAGAWGGAAGGAACACC Si Dn GWWCMAUCCCUGGGCUCAACCUAGWAWGCAU~CCWAAGACUAGAWAUWCAGAGCAAGGCGGAA~CCAWWAGCAGUGAAAUGCWAGAUA~GGAAGGAACACCDn Ec tAUWWMUCCCCGGGCUCAACCUGGtMCUGCAUCUCAUACUGGCAAGCUJ~~UCWAGAGGGGGWAtMWCCAG~WAGCG~GAAAUGCWAGAGAUCUGGAGGAAUACCEc 610 620 630 640 650 660 670 680 690 700 71 0 720 8 8 Ch CAUGGCWAGGCAGCUIUtUGGWCCAUACUGACGCUCallWGCGaAAGCWGGGGAGCAMCAGGAWAGAUACCCUGWAGUCCACGCCCUAAACGAUWCAACUAGGCGUCGGG~GCh Si CAUGGCGMGGCAGCCWCUGGGGCMUACUCACGCUCAUWGCGMAGCGUGGGUAGCAMCAGCAWAGAUACCCUGWAWCCACGCCCUAAACGAUWCMCUAGGUMGGGCAC Si Dn CAUGGCWAGGCAGCCUJ~GGGGCMUACUGACACUCAUWGCWAAGCGUGGGUAGCAAACAGCAWAWUACCCUGWAWCCACGCCCUAAACGAUWCAACUAGWGWGGGUA-Dn Ec GWCGCWAGGCGGCCCCCUGGACGAACACUCACGCUCAGWGCGAAAGCWGGGGAGCAMCAGCAWAGAUACCCUGWAWCCACGCCGUAAACGAUWCGACWGGAGGWGUG~C EC 730 740 ?50 760 770 780 790 800 810 820 830 840 r 8 IT Ch -UleMtACUCGWGCnCCAGCMCGC~CUlCACCGCCUGGG~WACGGCCGCMGWUWMCUCAMWMWCACGGGGACCCGCnCMGCGWGGAGCAUWG~AAUCh Si -WMWGtllCGWACCGCAG~MCGCAWM~CACCGCCUGGGCAWACGGCCGCMGWU~CUCMAWMWGACGGGGACCCGCACAAGCGWGGAGCAUGUG~AAUSi Dn -WM-UAtUCGWACCCMGCUMCGCUMCUlCACCGCCUGGGCAWACGGCCGCGCUl~CUCAM~WGACGGGGACCCGCACAAGCGWGGAGCAUWGGUAAU Dn Ec CUlCA6GCWGGCUJCCGGAGCUAACGCCUlMWC~CCGC~GGGGAWACGGCCGCGGUJ~CUCAMUGMW~CGGGGGCCCGCACAAGCGWGGAGCAUGUGG~AAUEC 85 0 860 a70 880 890 900 910 920 930 940 950 960 a Ch UCGAUGcMCGCGAA~CCWACCAGGCCUJCACAUCCUAGGMCMGGCAGACAUGCCWGWGCCWCGGCAACCUAGAGACAGWMGCAUGGCUWCGUCAgCUCWWCGUGA Ch Si UCCAUGcMCGCGAACMCCUIACCAGGCCWGACAUCCA~tM~ACCAGAGAUGWU1CWGCCUJCGGGAACUCUGAGACAGWG~GCAUGGCUWCWCAGCUCWGUCGUGASi Dn UCGAUtCMCGCGAAGAACCUIACCAGGCUUGACAUCCAGAGAAWCUACAGAGAUWGGGAWGCCUJCGGGAAtUCUGAGACACWGWGCAUGGCUWCGUCAGCUCGUGUCGUGA Dn Ec UCGAUGCMCGCGAAGAACCUJACCUGWCWGACAUCCACGGAACUUlUCAGAGAUGAGAAUWGCCUJCGGGAACCWGAGACAGGUGCUGCAUGGCUWCGUCAGCUCGUGUUGUGA EC 970 980 990 1000 1010 1020 1030 1040 1050 1060 1070 1080

Ch GAUWUGGGUJMWCCCGCMCCAGCGCMCCCWAUCCWA~GCCAGCAC~GG-WGGtMCUWMGCAGACUGCCGWGACAAACCGGAGGAAGWGGtGAUGACGUCAAGUCCh Si CAUCUlGCCUlMWCCCGCMCGACCCCMCCCUIAUC~ACUJGCCAGCACMCGG-WGGtMCUAUMGGAGACUGCCGWGACAAACCGGAGCAAGGUCGGGAUGACGUCAAGUC Si Dn GAUCUlCCGUJAAWCCCGCAACGAGCGCMCCCUlAUC~CUJGCCAGCACW~GWGGtMCUAUAAGGAGACUGCCGWtACAAACCGGAGGAAGGUGGGGAUGACGUCAAGUC On Ec AAUCUlG~GUUAAWCCCGCAACWGCGCAACCCUJAUCCUUGUJGCCAGCGWCCGG-CCGGCAACUCAAAGGACACUGCCAW~UAAACUGGAGGAAGWGGGGAUGACGUCAAGUC EC lop0 1100 1110 1120 1130 1140 1150 1160 1170 1180 1190 1200 IT Ch AUCAUGGCCCWACGGCCUGGGCUACACACWGCUACMUGWCGWACAGACGWAGCWAGCCGCCAGWGGAGCCMUCUWGAAAGCCGAUCWAGUCCGGAWCCAGUCUGCAAC Ch Si AUCAUGGCCCWACGGCCUGGGCUACACAC~G~ACM~~~CG~ACAGACG~GCCMCCCGCCAGGGGGAGCUAAUCUGAGAAAGCCGAUCGUAGUCCGGAWGCACUCUGCAACSi Dn AUCAUGCCCCUJACAGCCUGGGCUACACACWGCUACAAUGGGCG~ACAGACGWACCCAACCCGCGAGGGGCAGCCAAUCUtAGAGAAAACCtUUCGUAWCCGGAWGCACUCUGCAAC Dn Ec AUCAUGGCCCUJACGACCAGGGCUACACAC~UGCUACAAUGGCGCAUA~AAAGA~AAGCGACCUCGCGAGAGCAAGCGGACCUCAUAAAGUGCGUCGUAWCCGGAUUCGACUCUGCAACEc 1210 1220 1230 1260 1250 1260 1270 1280 1290 1300 1310 1320 a r Ch U~CACUGCAUGAAWCGGAAUCGCuAWAAucGCGAAuc~~~A~~~~~~~GA~~Ac~CccGGWCwWACACACUGCCCGUCACACCAUGGGA~GWGCaCCAGAAGCAGCUCh Si UCGAWGCAUGAAWCGGAAUCGCUAWAAUCGCGAAUCACMCWC~C~WCAAUACCU~CCCGGWCUJWACACACUGCCCGUCACACCAUGGGAWGGGWGCACCAGAAGUAGCU Si Dn UCWCUGCAUWAWCGCMUCGCUAWAAUCGCWAUCAGCACWCGCGWCAAUACWUCCCGGW~WACACACUGCCCWCACACCAUGGGA~~GCACCAGAAWAGGUDn Ec UC~CUCCAU~AWCG~AUCGCUAWAAUCWG~UCA~UCCCACGWGAAUAC~CCCGGGCCUJ~ACACACCGCCCGUCACACCAUGGGAWGGGWGCAAAACAA~UAGGUEc 1330 1340 1350 1m 1370 1380 1390 1400 1410 1420 1430 1460

Ch AGCUJMC .WaG- GAGGGCGCWGCCACGWWGGCCCAu ...... Ch Si AGCUJA- .-UCAG- ..-GGCGGWACCACGWWtWCA~aCUAGGG...... si Dn AG~MCCWAAGGAGGGCGCUlACCACGWWGGCC~UGACUGGGWGAAWCWAACAAGWAACCWAGGGGAACCUGCGGWGGAUCACCUCC~Dn Ec ~~~AACCUJCGGGAGGGCGCWACCACUWGAWCAUtACUG~GWGAAWCWAACAAGWAACCWACGGGAACCUGCGG~GGAUCACCUCCWAEc 1450 1460 1470 14sO 1490 1500 1510 1520 1530 lSC0 FIG. 1. Aligned sequences of members of the Cardiohacteriaceae. The sequences of Cardiobacteriurn horninis (Ch), Suttonella indologenes (Si), and Dichefobacter nodosus (Dn) are aligned with the sequence of Escherichia coli (Ec). The numbering system is the Escherichia coli numbering system (4). A and a, Adenine; C and c, cytosine; G and g, guanine; U and u, uracil; n, base not determined. Lower-case letters indicate some uncertainty in base identity. Dashes indicate gaps that were inserted for alignment of sequences, and dots indicate regions that were not sequenced. The Greek letters above the aligned sequences indicate a beta division Proteobacteria signature (p), a gamma division Proteohacteria signature (r),or a rare or unique signature for the Cardiobacteriaceae (a).

428 VOL. 40, 1990 CARDIOBACTERIACEAE FAM. NOV. 429

TABLE 2. Similarity matrix"

Ec Pv AL Pfn Hi RI 01 Pa Cv Ch Si On Ek Nd Kd Ng Kk Sv Rp Pc Af EC - 94.0 89.7 88.8 88.9 88.0 85.7 86.1 85.7 85.8 85.1 84.3 83.6 83.7 83.7 82.7 82.9 82.8 82.4 82.7 83.2 PV 6.3 - 89.2 87.4 87.8 87.0 86.3 85.1 84.9 84.7 84.5 83.5 83.4 83.7 83.7 83.3 83.6 84.0 83.0 82.5 82.7 AL 11.0 11.7 - 94.8 94.7 86.4 84.8 83.7 84.7 85.6 84.3 83.4 82.7 82.7 82.7 81.3 82.1 82.7 82.3 82.0 82.3 Pm 12.1 13.8 5.4 - 95.7 86.4 84.7 83.7 84.3 85.3 83.8 82.9 82.0 82.7 82.7 81.2 82.0 82.8 82.5 81.9 82.0 Hi 12.0 13.3 5.5 4.4 - 87.1 64.5 86.3 84.5 66.2 84.8 64.2 82.7 82.8 83.4 81.3 81.9 83.1 82.8 82.0 81.9 Ra 13.1 16.2 15.1 15.0 14.1 - 65.7 86.3 85.0 85.8 84.1 84.3 82.2 81.6 82.4 82.0 82.3 83.4 82.6 81.7 82.7 01 15.9 15.2 17.0 17.2 17.6 15.8 - 89.3 86.2 85.9 85.2 83.3 82.7 63.1 83.6 82.7 82.7 84.9 83.3 84.8 83.6 Pa 15.3 16.6 18.4 18.4 17.7 15.2 11.6 - 88.0 87.4 86.8 85.6 84.7 85.0 84.9 83.7 83.5 84.8 84.2 04.4 84.6 CV 15.8 16.9 17.1 17.6 17.4 16.8 15.5 15.0 - 88.7 66.4 84.8 85.3 64.9 85.7 84.4 84.7 66.1 84.9 85.7 85.7 Ch 15.7 17.2 15.7 16.4 15.3 15.7 15.6 13.8 12.2 - 92.9 92.0 85.6 85.8 66.0 85.3 84.2 85.0 85.6 84.9 84.4 Si 16.6 17.4 17.6 18.2 17.0 17.9 16.4 14.5 15.0 7.4 - 94.1 85.9 65.8 66.5 85.9 85.2 85.1 84.5 84.7 84.2 On 17.6 18.6 18.7 19.4 17.7 17.7 18.8 16:O 17.0 8.5 6.1 - 84.7 64.3 84.7 84.6 83.7 84.7 84.0 84.0 83.7 Ec 18.5 18.7 19.7 20.6 19.6 20.3 19.7 17.1 16.4 16.0 15.6 17.2 - 97.5 97.6 95.3 95.3 88.2 88.0 90.2 90.0 Nd 16.3 18.3 19.7 19.7 19.5 21.1 19.1 16.8 16.9 15.7 15.8 17.7 2.6 - 97.1 95.9 95.5 88.5 88.7 89.9 90.0 Kd 18.3 18.4 19.6 19.6 18.7 20.0 18.5 16.9 15.9 15.5 14.9 17.1 2.4 2.9 - 95.6 95.8 88.4 87.9 89.5 89.5 Ng 19.6 18.8 21.6 21.7 21.6 20.5 19.7 18.3 17.5 16.4 15.7 17.3 4.9 4.2 4.5 - 94.8 87.6 88.0 69.4 88.4 Kk 19.4 18.5 20.4 20.5 20.7 20.2 19.6 18.6 17.2 17.7 16.5 18.4 4.9 4.6 4.3 5.4 - 88.5 88.1 89.8 88.9 Sv 19.5 17.9 19.6 19.5 19.1 18.7 16.9 17.0 15.3 16.7 16.6 17.1 12.8 12.5 12.6 13.6 12.5 - 90.3 90.4 89.0 Rp 20.0 19.3 20.1 19.9 19.5 19.8 18.9 17.7 16.9 16.0 17.4 17.9 13.1 12.3 13.2 13.0 12.9 10.4 - 90.7 89.3 PC 19.7 19.9 20.5 20.7 20.6 20.9 17.0 17.5 15.9 16.9 17.1 17.9 10.5 10.8 11.3 11.4 10.9 10.3 9.9 - 90.7 Af 19.0 19.6 20.2 20.6 20.7 19.6 18.5 17.3 15.9 17.5 17.7 18.3 10.8 10.8 11.3 12.6 12.0 11.9 11.6 9.9 -

a Abbreviations: Ec, Escherichia coli; Pv, Proteus vulgaris; Al, Actinobacillus lignieresii; Pm, Pasteurella multocida; Hi, Haemophilus injluenzae; Ra, Ruminobacter amylophilus; 01, Oceanospirillum linum; Pa, Pseudomonas aeruginosa; Cv, Chromatium vinosum; Ch, Cardiobacterium hominis; Si, Suttonella indologenes; Dn, Dichelobacter nodosus; Ec, Eikenella corrodens; Nd, Neisseria denitrijicans; Kd, Kingella denitrificans; Ng, Neisseria gonorrhoeae; Kk, Kingella kingue; Sv, Spirillum volutans; Rp, Rhodocyclus purpura; Pc, [Pseudomonas] cepacia; Af, Alcaligenes fuecalis. The numbers above the diagnoal are uncorrected percentages of similarity. The numbers below the diagonal are percentages of difference corrected for multiple base changes by the method of Jukes and Cantor (17). should be transferred to the new genus Suttonella, which is diameter and 2 to 3 p,m long and have rounded ends. Cells named after the Australian microbiologist R. G. A. Sutton, occasionally occur in pairs, clusters, or chains. Gram-nega- who was one of the first workers to describe a strain of this tive, but there is a tendency to resist Gram decolorization. species. Nonmotile as determined by normal tests, but may be Description of the genus Suttonella. Suttonella (Sut. fimbriated (pileated) and exhibit twitching motility. Small ton.el'la. L. dim. ending ella; N.L. fem. n. Suttonella, translucent colonies grow on blood agar after 48 h of named after R. G. A. Sutton). The description of the genus incubation at 37°C. Pitting of media and spreading edges of Suttonella below is based on the description of Snell and colonies may be present with freshly isolated strains, but Lapage (36), supplemented with information from Bovre et may be lost. Aerobic. Oxidase positive (when tested with al. (3) and Bruun et al. (5). Straight rods that are 1.0 p,m in tetramethyl-p-phenylene diamine). Catalase negative. Indole

0.05 Escherichia coli Proteus vulgaris ,-. ,-. Actfnobacillus lignieresii Pas t eur el la mul t oci da Haemopbi lus in f luenrae ffuminobacter amylophilus Oceanospirillum lfnum Pseudomonas aeruginosa Cbromat ium vinosum Cardiobacterium bominf s Suttonella indologenes Dfcbelobacter nodosus €ikenella corrodens vNefsseria deni tri ficans Kingella deni trf ficans Nefsseria gonorrhoeae Kingella kfngae Spiri llum volutans ffhodocyc I us purpura /Pseudomonas] cepac i a A lcal igenes f aeca l i s FIG. 2. Phylogenetic tree for the beta and gamma divisions of Proteobacteria. The scale bar represents a 5% difference in nucleotide sequences as determined by measuring the lengths of the horizontal lines connecting two species. 430 DEWHIRST ET AL. INT. J. SYST.BACTERIOL.

TABLE 3. Phenotypic characteristics of members of by the triple sugar iron method). No growth occurs on the family Cardiobacteriaceae MacConkey agar. The G+C content of the DNA is 49 mol%. Cardio- Suttonella Dichelo- The genus contains a single species, Suttonella indologenes. Characteristic bacterium indolo- bacter Description of Suttonella indologenes (Snell and Lapage hominis genes nodosus 1976). Suttonella indologenes (in.do1.o’ge.nes. N.L. n. in- dolum, indole; N. L. suff. genes, producing; N.L. adj. Rods +a + + Cell width (pm) 0.5-0.75 1 1-1.7 indologenes, indole producing) (synonyms, ‘‘Bijsterveldl Cell length (pm) 1-3 2-3 3-6 Sutton” strains van Bijsterveld 1970, Sutton, O’Keefe, Gram negative but resists decol- + + + Bundock, Jeboult, and Tester 1972, Kingella indologenes orization Snell and Lapage 1976). The description below is based on Pili (N-methylphenylalanine ND + + the description of Snell and Lapage (36), supplemented with type) information from Bruun et al. (5) and Wallace et al. (43). Crystalline surface layer + ND + Straight rods that are 1.0 pm in diameter and 2 to 3 pm long Intrusive membranes + ND + and have rounded ends. Cells occasionally occur in pairs, Twitching motility + + + rosette clusters, or chains. Gram negative, but there is a Colonies with spreading edges + + + tendency to resist Gram decolorization. Nonmotile as deter- Pitting and corrosion of agar ND + + Aerobic + + - mined by normal tests, but may be fimbriated (pileated) and Catalase activity - - - exhibit twitching motility. Aerobic. Aerobic growth is en- Oxidase activity + + - hanced by high humidity and CO,, which makes the organ- Indole production + + - isms appear to be facultatively anaerobic, Oxidase positive Alkaline phosphatase activity - + + (W) (when tested with tetramethyl-p-phenylene diamine). Cata- Urease activity - - - lase negative. Indole positive (activity may be weak). Urease DNase activity - - - - negative. DNase negative. Ornithine and lysine decarboxyl- Trypsin + + ase negative, Alkaline phosphatase positive. Trypsin (N-ben- Phosphohydrolase activity + - ND Lysine decarboxylase activity - - - zoyl-~~-arginine-2-naphthylamide)negative. Phosphohydro- Arginine decarboxylase activity - - - lase (naphthyl-AS-B1-phosphate) negative. Chemoorgano- Ornithine decarboxylase activity - - + trophic, having a strictly fermentative type of metabolism. p-Galactosidase activity - - ND Acid but not gas is produced from fructose, glucose, malt- y-Glutamyltransferase acitivity + + ND ose, mannose, and sucrose. The maltose reaction is weak Reduction of and delayed (positive at 28 days) but is negative as deter- Nitrate - - - mined by rapid (4- to 24-h) protocols. No acid is produced Nitrite d + ND from adonitol, arabinose, cellobiose, dulcitol, ethanol, ga- Resazurin + + ND lactose, inositol, lactose, melibiose, raffinose, rhamnose, Selenite ND ND + Acid produced from: salicin, trehalose, or xylose. Nitrates are not reduced. Ni- Glucose, sucrose, maltose, + + - trites are reduced by some strains. Resazurin is reduced. mannose, or fructose H,S is produced (as determined by the lead acetate method, Sorbitol or mannitol + - - but not by the triple sugar iron method). No growth occurs Arabinose, xylose, rhamnose, - - - on MacConkey agar. The major cellular fatty acids are as galactose, lactose, trehalose follows: 18:l (o~c),27%; 16:0, 26%; 16:l (o~c),19%; 14:0, melibiose, cellobiose, ra%- 11%; 12:0,5%; and 3-OH 12:0,2%. The G+C content of the nose, salicin, adonitol, dulci- DNA is 49 mol%. The type strain is strain ATCC 25869. The to1 inositol, or ethanol description of the type strain does not differ from the Ammonia produced from: Arginine , asparagine, serine , ND ND + description of the species. or threonine Evidence that [Bacteroides]nodosus is not a member of Phenylalanine, cystine, citrul- ND ND - the genus Bacteroides has been accumulating over the past line, or ornithine 12 years. Stewart reported in 1977 (37) that the lipopolysac- Hydrolysis of charide of [Bacteroides] nodosus contains 2-keto-3-deoxy- Gelatin - - + octonic acid and heptose, whereas 2-keto-3-deoxyoctonic Casein - + + acid and heptose are absent from other true Bacteroides Albumin ND ND + species (13). [Bacteroides]nodosus does not possess mena- Meat ND ND + quinones which are found in other Bacteroides species (30). Tween 20 - + ND Tween 40 - + ND Johnson and Harich (16) demonstrated by using RNA-DNA Tween 80 - - + homology studies that [Bacteroides]nodosus has an average H,S production (lead acetate + + + level of homology of less than 20% with other species of paper) Bacteroides. Finally, partial sequencing of the 16s rRNA G+C content (mol%) 59-60 49 45 gene (19) indicated that [Bacteroides]nodosus is a member of the Proteobacteria division rather than the Flavobacte- a +, Positive; -, negative; w, weak; d, different reactions in different strains; ND, not determined. rium-Bacteroides division of eubacteria as defined by Woese (46) and Woese et al. (47) and therefore must be transferred to a different genus. Because [Bacteroides] nodosus is phe- positive (activity may be weak). Urease negative. DNase notypically and genetically distinct from the genera Cardio- negative. Ornithine and lysine decarboxylase negative. bacterium and Suttonella, we propose that [Bacteroides] Chemoorganotrophic, having a strictly fermentative type of nodosus should be transferred to a new genus, Dichelobac- metabolism. Acid but not gas is produced from glucose and ter, as Dichelobacter nodosus. a limited number of sugars. Nitrates are not reduced. H,S is Description of the genus Dichelobacter. Dichelobacter produced (as determined by the lead acetate method, but not (Di.che’1o.bac.ter. Gr. adj. Dichelos, cloven hoofed; N. L. VOL. 40, 1990 CARDIOBACTERIACEAE FAM. NOV. 431

n. bacter, a rod; N. L. masc. n. Dichelobacter, cloven- Scientific and Industrial Research Organization, McMaster hoofed rod, because this organism is the rod-shaped bacte- Laboratory, New South Wales, Australia, has been used as rium that causes footrot in sheep, goats, and cattle). The the reference strain for the organism (1, 8, 11, 12, 37,38), we description below is based on data published previously by formally propose that this strain be designated a reference Beveridge (2), Cat0 et al. (7), Holdeman et al. (14), and strain. Strain 198A was isolated by J. R. Egerton from a Skerman (33). Large straight or slightly curved rods that are sheep at Wollogorang Station, Goulburn, New South Wales, 1.0 to 1.7 pm in diameter and 3 to 6 pm long and have Australia, in the late 1960s (J. R. Egerton, personal commu- rounded ends. The cells often have terminal swellings, nication). Strain 198A (=VPI 5731-1) was sent to L. V. although this feature is less pronounced in subcultured Holdeman at Virginia Polytechnic Institute and State Uni- organisms. Gram negative, but there is a tendency to resist versity, Blacksburg, who deposited it with the American decolorization. Organisms stained with Loeffler methylene Type Culture Collection, Rockville, Md., as ATCC strain blue have prominent polychromic granules toward the poles 27521. Type strain ATCC 25549 (=VPI 2340) was also and at intermediate sites within the cells. The cells have deposited by L. V. Holdeman, who obtained it from L. DS. large numbers of pili (N-methylphenylalanine-typepilins) Smith (L. DS. Smith strain 11342), who obtained it from which vary in number according to changes in colony W. I. B. Beveridge (7). The description of the origin of strain morphology. Cells exhibit twitching motility, and spreading ATCC 25549T in Bergey ’s Manual of Systematic Bacteriol- of colonies occurs. Ammonia is produced from arginine, ogy appears to be in error as it suggests that strain ATCC asparagine, serine, and threonine. Positive for ornithine 25549T was derived from strain 198A (14). decarboxylase phosphatase (weak), H,S production, selen- The level of 16s rRNA similarity among the genera ite reduction, and proteolytic activity on gelatin, casein, and Cardiobacterium, Suttonella, and Dichelobacter (93%) sug- albumin. Negative for acid or gas production from carbohy- gests that these taxa should be placed in a single family. drates, ammonia production from phenylalanine, cysteine, However, Cardiobacterium hominis and Suttonella indolo- citrulline, and ornithine, starch hydrolysis, esculin hydroly- genes are aerobic and saccharolytic, while Dichelobacter sis, indole production, nitrate reduction, growth in 0.1% nodosus is anaerobic and asaccharolytic. These differences bile, hemolysis, arginine decarboxylase, catalase , oxidase, pose the diacult taxonomic problem of what range of urease, DNase, coagulase, lipase, lecithinase, and hyalu- phenotypic difference can be tolerated within a genus, ronidase. The G+C content of the DNA is 45 mol%. The family, or larger taxonomic unit. This problem is being genus contains a single species, Dichelobacter nodosus. encountered with increasing frequency as ribosomal cistron Description of Dichelobacter nodosus comb. nov. (Beveridge analysis and 16s rRNA sequencing studies show that organ- 1941). Dichelobacter nodosus (no.do’sus. L. adj. nodosus, isms with major phenotypic differences are closely related full of knots, referring to the shape of the cells) (synonyms, genomically. For example, the facultatively anaerobic or- “organism K” Beveridge 1938, Fusiformis nodosus Bever- ganism Eikenella corrodens is clearly a member of the idge 1941, Ristella nodosus (Beveridge 1941) PrCvot 1948 aerobic family Neisseriaceae (10, 28). In addressing this [26], Bacteroides nodosus (Beveridge 1941) MrAz 1963 [23]). problem, we were guided by the advice presented by Wayne The description below is based on data published previously et al. (44). While Wayne et al. offer general guidelines, they by Beveridge (2), Cat0 et al. (7), Holdeman et al. (14), and do not give any specific guidance for resolving the problem Skerman (33). Large straight or slightly curved rods that are raised by our findings. Members of the Ad Hoc Committee 1.0 to 1.7 pm in diameter and 3 to 6 pm long and have on Reconciliation of Approaches to Bacterial Systematics rounded ends. The cells often have terminal swellings, raised the concern that any phylogenetically based taxo- although this feature is less pronounced after repeated nomic scheme should show phenotypic consistency (44). We transfers. Gram negative, but there is a tendency to resist also believe that genera should be phenotypically coherent. decolorization. Organisms stained with Loeffler methylene We suggest that regardless of how close two species are blue have prominent polychromic granules toward the poles genetically, if there are major phenotypic differences, then and at intermediate sites within the cells. Cells have large the species should be placed in separate genera. The func- numbers of pili (N-methylphenylalanine-typepilins) which tioning of clinical microbiology demands and deserves phe- vary in number according to changes in colony morphology. notypic consistency at the genus level. However, if nomen- Cells exhibit twitching motility, and spreading of colonies clature “is to reflect the genomic relationships to the greatest occurs. Ammonia is produced from arginine, asparagine, extent possible” (44), then at some point genomically related serine, and threonine. Positive for ornithine decarboxylase, but phenotypically disparate organisms must be placed phosphatase (weak), H,S production, selenite reduction, within a single taxonomic unit. We suggest that the family is and proteolytic activity on gelatin, casein, and albumin. a sufficiently robust taxonomic unit for this purpose. Our Negative for acid or gas production from carbohydrates, approach to the description of a family with phenotypically ammonia production from phenylalanine, cysteine, citrul- disparate genera is to base the description on the phenotyp- line, and ornithine, starch hydrolysis, esculin hydrolysis, ically most robust genera. In general, major phenotypic indole production, nitrate reduction, growth in 0.1% bile, traits, such as aerotolerance, motility, photosynthesis, and hemolysis, arginine decarboxylase, catalase, oxidase, ure- saccharolytic ability, are lost more easily (occasionally ase, DNase, coagulase, lipase, lecithinase, and hyaluroni- through a single mutation) than they are acquired by evolu- dase. The G+C content of the DNA of Skerman strain 10 is tion or horizontal transfer. We hope that the description of 45 mol%. The type strain is strain ATCC 25549. The descrip- the new family Cardiobacteriaceae below demonstrates a tion of type strain ATCC 25549 (7) is similar to the descrip- successful reconciliation of phenotypic and phylogenetic tion of the species. The growth of most strains on an approaches to bacterial systematics. appropriate medium, such as TAS medium, is more luxuri- Description of Cardiobacteriaceaefam. nov. Cardiobacteri- ant than the growth described for strain ATCC 25549T (T = aceae (Car.di.o.bac. te.ri.a’ce.ae. N. L. n. Cardiobacterium, type strain). bacterium of the heart; aceae, ending to denote a family; Because in essentially all previously published work on N.L. fem. pl. n. Cardiobacteriaceae, the family of cardio- [Bacteroides] nodosus strain 198A from Commonwealth bacteria). Straight rods that are 0.5 to 1.7 pm in diameter and 432 DEWHIRST ET AL. INT. J. SYST. BACTERIOL.

TABLE 4. Traits that differentiate the genera of the Cardiobacteriaceae from phenotypically similar taxa

Oxidase Catalase Indole Fermentation of Nitrate y-Glutamyltrans- Taxon activity activity production Glucose Sorbitol reduction ferase activity

Cardiobacterium + Suttonella + Dichelobacter ND Kingella - Eikenella corrodens - Neisseria D Actinobacillus actinomycetemcomitans + Haemophilus aphrophilus + Pasteurella D

a +, Positive; -, negative; D, variable within the taxon; ND, not determined.

1 to 6 pm long and have rounded ends. Cells occasionally Lautrop, and J. J. S. Snell. 1974. Studies on a collection of occur in pairs, rosette clusters, or chains. Gram negative, gram-negative bacterial strains showing resemblance to mor- but there is a tendency to resist Gram decolorization. axellae: examination by conventional bacteriological methods. Nonmotile as determined by normal tests, but may exhibit Int. J. Syst. Bacteriol. 24:438-446. 4. Brosius, J., M. L. Palmer, P. J. Kennedy, and H. F. Noller. 1978. twitching motility and possess N-me t hylphen ylalanine- type Complete nucleotide sequence of a 16s ribosomal RNA gene pilins. Aerobic and oxidase positive except for members of from Escherichia coli. Proc. Natl. Acad. Sci. USA 75: the anaerobic genus Dichelobacter. Catalase negative. Ure- 4801-4805. ase negative. Fermentative type of chemoorganotrophic 5. Bruun, B., Y. Ying, E. Kirkegaard, and W. Frederiksen. 1984. metabolism except for members of the genus Dichelobacter, Phenotypic differentiation of Cardiobacterium hominis, Kin- which are arginine chemoorganotrophs. Acid but not gas is gella indologenes and CDC group EF-4. Eur. J. Clin. Microbiol. produced from glucose, sucrose, maltose, mannose, and 3:230-235. fructose in fermentative species. Nitrates are not reduced. 6. Carbon, P., J. P. Ebel, and C. Ehresmann. 1981. The sequence H,S is produced (as determined by the lead acetate method). of the ribosomal 16s RNA from Proteus vulgaris. Sequence comparison with E. coli 16s RNA and its use in secondary No growth occurs on MacConkey agar. The C+C content of structure model building. Nucleic Acids Res. 9:2325-2333. the DNA is 45 to 60 mol%. Species in the family possess the 7. Cato, E. P., L. V. Holdeman, and W. E. C. Moore. 1979. following rare or unique signatures in their 16s rRNAs: Proposal of neotype strains for seven non-saccharolytic Bac- uracil at positions 18, 1051, and 1400 and adenine at position teroides species. Int. J. Syst. Bacteriol. 29:427-434. 917 (E. coli numbering). The family comprises the following 8. Claxton, P. D., L. A. Ribeiro, and J. R. Egerton. 1983. Classi- three genera: Cardiobacterium (the type genus), Suttonella, fication of Bacteroides nodosus by agglutination tests. Aust. and Dichelobacter. Vet. J. 60:331-334. The major phenotypic traits which distinguish the genera 9. De Vos, P., A. Van Landschoot, P. Segers, R. Tytgat, M. Gillis, of the Cardiobacteriaceae from each other and from pheno- M. Bauwens, R. Rossau, M. Goor, M. Pot, K. Kersters, P. typically related species and genera are shown in Table 4. Lizzaraga, and J. De Ley. 1989. Genotypic relationships and taxonomic localization of unclassified Pseudomonas and Pseu- Cardiobacteriurn hominis and Suttonella indologenes are domonas-like strains by deoxyribonucleic acid-ribosomal ribo- distinguished most easily by the following traits: Cardiobac- nucleic acid hybridizations. Int. J. Syst. Bacteriol. 39:35-49. terium hominis is positive for trypsin, phosphohydrolase, 10. Dewhirst, F. E., B. J. Paster, and P. L. Bright. 1989. Chromo- sorbitol, and mannitol acidification, whereas Suttonellu in- bacterium, Eikenella, Kingella, Neisseria, Simonsiella, and dologenes is positive for alkaline phosphatase and Tween 20 Vitreoscilla species comprise a major branch of the beta group and Tween 40 hydrolysis. Proteobacteria by 16s ribosomal ribonucleic acid sequence comparison: transfer of Eikenella and Simonsiella to the family ACKNOWLEDGMENTS Neisseriaceae (emend.). Int. J. Syst. Bacteriol. 39:258-266. 11. Elleman, T. C., and P. A. Hoyne. 1984. Nucleotide sequence of We acknowledge Robert Cromey , Virginia Commonwealth Uni- the gene encoding pilin of Bacteroides nodosus, the causal versity, and Thomas 0. MacAdoo, Virginia Polytechnic Institute organism of ovine footrot. J. Bacteriol. 160:1184-1187. and State University, for their assistance in naming the organisms. 12. Every, D., and T. M. Skerman. 1980. Ultrastructure of the We also thank John R. Egerton, University of Sydney, for helpful Bacteroides nodosus cell envelope layers and surface. J. Bac- discussions. teriol. 141:845-857. This work was supported by Public Health Service grants DE- 13. Hofstad, T. 1974. The distribution of heptose and 2-keto-3- 02847, DE-04881, and DE-08303 from the National Institutes of deoxy-octonate in Bacteroidaceae. J. Gen. Microbiol. 85: Health and by a grant from the Australian Wool Corporation. S.L. 314-3 20. is the recipient of an Australian Wool Corporation Postgraduate 14. Holdeman, L. V., R. W. Kelley, and W. E. C. Moore. 1984. Scholarship. Genus 1. Bacteroides Castellani and Chalmers 1919, 959*? p. 604-631. In N. R. Krieg and J. G. Holt (ed.), Bergey's manual LITERATURE CITED of systematic bacteriology, vol. 1. The Williams & Wilkins Co., Anderson, B. J., C. L. Kristo, J. R. Egerton, and J. S. Mattick. Baltimore. 1986. Variation in the structural subunit and basal protein 15. Jenny, D. B., P. W. Letendre, and G. Iverson. 1987. Endocardi- antigens of Bacteroides nodosus fimbriae. J. Bacteriol. 166: tis caused by Kingella indologenes. Rev. Infect. Dis. 9:787-789. 453-460. 16. Johnson, J. L., and B. Harich. 1986. Ribosomal ribonucleic acid Beveridge, W. I. B. 1941. Foot-rot in sheep: a transmissible homology among species of the genus Bacreroides. Int. J. Syst. disease due to infection with Fusiformis nodosus (n.sp.). Bull. Bacteriol. 36:71-79. Sci. Ind. Res. (Melbourne) 14O:l-53. 17. Jukes, T. H., and C. R. Cantor. 1969. Evolution of protein Bovre, K., J. E. Fuglesang, S. D. Henriksen, S. P. 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