Achromobacter Xylosoxidans (Yabuuchi and Yano) and Proposal of Achromobacter Ruhlandii (Packer and Vishniac) Comb

Microbiol. Immunol., 42(6), 429-438, 1998

Emendation of Genus Achromobacter and Achromobacter xylosoxidans (Yabuuchi and Yano) and Proposal of Achromobacter ruhlandii (Packer and Vishniac) Comb. Nov., Achromobacter piechaudii (Kiredjian et al.) Comb. Nov., and Achromobacter xylosoxidans Subsp. denitrificans (Roger and Tan) Comb. Nov.

Eiko Yabuuchi*,', Yoshiaki Kawamura2, Yoshimasa Kosako3, and Takayuki Ezaki2

'Departmentof Microbiologyand Immunology , AichiMedical University,Faculty of Medicine,Aichi 480-1195, Japan, 'Depart- ment of Microbiology, Gi;fu UniversityMedical School, Gifu, Gifu 500-8705, Japan, and'Institute of Physical and Chemical Research, RIKEN, Wako,Saitama 351-0198, Japan

ReceivedMarch 9, 1998. AcceptedMarch 25, 1998

Abstract: Based on the results of GC content determination and 16S rRNA sequence analysis among the type strains of Achromobacter xylosoxidans, 4 Alcaligenes species, 5 Bordetella species, and 12 species of 4 other genera, the separation of genus Achromobacter Yabuuchi and Yano 1981, with the type species Achro- mobacter xylosoxidans, is confirmed. Alcaligenes ruhlandii (Packer and Vishniac) Aragno and Schlegel 1992 is a distinct species and not a senior synonym of Achromobacter xylosoxidans. Alcaligenes ruhlandii and Alcaligenes piechaudii Kiredjian et al 1986 are transferred to genus Achromobacter. Thus 2 new combinations, Achromobacter ruhlandii (Packer and Vishniac) and Achromobacter piechaudii (Kiredjian et al) are proposed; their type strains are ATCC 15749 and ATCC 43552, respectively. Alcaligenes denitrificans Riiger and Tan 1983 is also transferred to genus Achromobacter and ranked down to the subspecies of Achro- mobacter xylosoxidans. Thus a new subspecies name, Achromobacter xylosoxidans subsp. denitrificans (Riiger and Tan) is proposed. The type strain of the subspecies is ATCC 15173. This proposal automatically creates type subspecies, Achromobacter xylosoxidans subsp. xylosoxidans, with type strain ATCC 27061. An emended description of genus Achromobacter and of type species Achromobacter xylosoxidans are given.

Key words: Achromobacter xylosoxidans, Achromobacter ruhlandii, Achromobacter piechaudii, Achro- mobacter xylosoxidans subsp. denitrificans

The species name Achromobacter xylosoxidans was from human ear discharge. More than 70 characters of first proposed in 1971 (24) for the 7 strains isolated 55 strains of A. xylosoxidans, including type strain for the

*Address correspondence to Dr. Eiko Yabuuchi, Omiya 4-19- Japan; GC, Guanine plus cytosine; GIFU, Department of Micro- 18, Asahi-ku, Osaka, Osaka 535-0002, Japan. E-mail: Achei- biology, Gifu University School of Medicine, Gifu, Japan; IAM, 08 @fa2.so-net.or. jp Institute of Applied Microbiology, University of Tokyo, Tokyo, Japan; JCM, Japan Collection of Microorganisms, RIKEN, Saita- Abbreviations: ATCC, American Type Culture Collection, ma, Japan; LMG, Laboratorium voor Microbiologie, Universiteit Maryland, USA; CCM, Czech Collection of Microorganisms, Gent, Gent, Belgium; NAD, nicotinamide adenine dinucleotide; Masaryk University, Brno, Tvrdeho, Czech Republic; CDC, Cen- ND, no datum; NCIB, National Collection of Industrial Bacteria, ters for Disease Control and Prevention, Georgia, USA; comb. Aberdeen, Scotland; NCTC, National Collection of Type Cul- nov., combinatio nova=new combination; Cu, copper; DSM, tures, London, England; NJ, neighbor-joining; OF, oxidation-fer- Deutsche Sammlung von Mikroorganismen and Zellkulturen, mentation; RH, Rudolph Hugh, Department of Microbiology, Braunschweig, Germany; EY, Eiko Yabuuchi, Department of George Washington University School of Medicine, Washing- Microbiology and Immunology, Aichi Medical University, Aichi, ton, D.C., USA; SSC, saline-sodium citrate; subsp., subspecies.

429 430 E. YABUUCHI ET AL species (ATCC 27061), were described in 1974 (25). ferred the species to genus Alcaligenes and proposed a Because the Judicial Commission has been asked by new combination Alcaligenes ruhlandii (Packer and Hendrie et al (10) to reject the genus name Achro- Vishniac) for the peritrichous hydrogen bacterium. They mobacter on the basis that no type strain of the type later described that an NAD-specific soluble hydroge- species Achromobacter liquefaciens exists, the com- nase, not accompanied by a membrane-bound hydro- mission agreed (11) at a meeting on 3 September 1978 to genase (2), was encountered in both A. ruhlandii and A. omit the name from the Approved Lists of Bacterial xylosoxidans. They referred to Alcaligenes ruhlandii and Names (20). Thus the genus name Achromobacter was Alcaligenes denitrificans as the synonyms of Alcali- not included in the Approved Lists and lost its standpoint genes xylosoxidans Kiredjian et al (vide p. 370). Kersters in nomenclature. However, the name was not included and De Ley (13) also treated A. ruhlandii as a synonym in the list of rejected generic names. In accordance of "Alcaligenes denitrificans subsp. xylosoxidans." If A. with Rule 28a and Provisional Rule B3 of the Bacterio- xylosoxidans and A. ruhlandii are synonymous, the spe- logical Code (1975 Revision) (7), Achromobacter cific epithet of the former species must be ruhlandii, xylosoxidans (ex Yabuuchi and Ohyama 1971) was which precedes over xylosoxidans. revived for the same organism to which the name was Based on phenotypic characteristics, DNA base com- originally applied, and the new name Achromobacter, not position, DNA-DNA similarity, and 16S rRNA sequence "Achromobacter ," Bergey et al 1923 was proposed in analysis, the emendation of Achromobacter as a separate which A. xylosoxidans was placed as the type species. genus and its type species Achromobacter xylosoxidans The strain ATCC 27061 was designated as the type are described, and three new combinations, Achro- strain of A. xylosoxidans (26). mobacter ruhlandii, Achromobacter piechaudii, and Alcaligenes denitrificans (Jensen) Monias 1928 was Achromobacter xylosoxidans subsp. denitrificans, are listed in the Index Bergeyana as a validly published proposed. legitimate name (3). Although strain NCTC 8582 was designated as the type for the species, Alcaligenes de- Materials and Methods nitrificans Leifson and Hugh 1954 (15) was illegitimate because the specific epithet denitrificans is the later Bacterial strains. Histories and corresponding num- homonym within the same genus (Rule 25d). Hendrie, bers of the 6 type strains for 6 species of 3 genera are list- Holding, and Shewan in 1974 (10) erroneously consid- ed in Table 1. The strains were inoculated on Bacto-heart ered A. denitrificans Leifson and Hugh as a subjective infusion (HI) agar and incubated at 37 C unless otherwise synonym of Alcaligenes faecalis Castellani and Chalmers stated. (5), and they requested the Judicial Commission to issue Base composition of DNA. DNA base composition of an Opinion to reject the name A. denitrificans. A. deni- Alcaligenes ruhlandii EY 391 8Twas determined by high trificans was excluded from the Approved Lists. On the performance liquid chromatography as described previ- basis of DNA base composition, DNA homology, and ously (9). The calculation of the mol% GC was based on nitrate reduction, Ruger and Tan revived the name the Escherichia coli K-12 strain DNA as a standard Alcaligenes denitrificans in 1983 as a separate new (51.1 mol% GC). species (18). DNA-DNA hybridization. DNA-DNA similarities In 1984, Kersters and De Ley transferred Achro- among the type strains of Achromobacter xylosoxidans, mobacter xylosoxidans to genus Alcaligenes and pro- Alcaligenes denitrificans, Alcaligenes ruhlandii, Alcali- posed two new subspecies, A. denitrificans subsp. deni- genes piechaudii, and Bordetella bronchiseptica were trificans and A. denitrificans subsp. xylosoxidans (13). determined by quantitative microplate DNA-DNA Because the epithet xylosoxidans has precedence over hybridization as described previously (8). DNAs from 6 denitrificans, the latter subspecific name was announced type strains were reassociated with biotin-labeled DNAs as illegitimate (16). In 1986, Kiredjian et al (14) cor- from the type strains of Achromobacter xylosoxidans rected these subspecies names as Alcaligenes xylosoxi- and Alcaligenes ruhlandii at 51 C (stringent condition) by dans subsp. xylosoxidans and Alcaligenes xylosoxidans using 2 X SSC and 50% formamide. subsp. denitrificans. The 16S rRNA gene sequences and their phylogenet- Ruhland (19) described hydrogen-oxidizing autotroph- ic analyses. The 16S rRNA gene sequences of the type ic bacteria and named them as Knallgasbakterien. In strains of Alcaligenes ruhlandii and Alcaligenes 1955, Packer and Vishniac (17) proposed the new name piechaudii from positions 45 to 1380 (E. coli numbering) Hydrogenomonas ruhlandii for a facultatively autotroph- were determined by the method described previously ic hydrogen-oxidizing (Knallgas) bacterium, without (12), and the results were deposited into the DNA Data designating type strain. Aragno and Schlegel (1) trans- Bank of Japan under accession numbers AB010840 and GENUS ACHROMOBACTER, EMENDATION 431

Table 1. Histories and corresponding number of 6 type strains of Achromobacter, Alcaligenes, and Bordetella species

ATCC, American Type Culture Collection, Maryland, USA; DSM, Deutsche Sammlung von Mikroorganismen and Zellkulturen, Braun- schweig, Germany; EY, Eiko Yabuuchi, Department of Microbiology and Immunology, Aichi Medical University, Aichi, Japan; GIFU, Department of Microbiology, Gifu University School of Medicine, Gifu, Japan; IAM, Institute of Applied Microbiology, University of Tokyo, Tokyo, Japan; JCM, Japan Collection of Microorganisms, RIKEN, Saitama, Japan; LMG, Laboratorium voor Micro- biologic Universiteit Gent, Gent, Belgium; NCIB, National Collection of Industrial Bacteria, Aberdeen, Scotland; NCTC, National Col- lection of Type Cultures, London, England; RH, Rudolph Hugh, Department of Microbiology, George Washington University School of Medicine, Washington, D.C., USA. "' Harumi Takeda , Clinical Microbiology Laboratory, Kansai Medical University, Moriguchi, Osaka, Japan.

Table 2. Sources of 16S rRNA nucleotide sequence data

* Weyant et al , Validation List No. 54. Int. J. Syst. Bacteriol. 45: 619, 1995. ** Spring et al , Validation List No. 61. Int. J. Syst. Bacteriol. 47: 601, 1996.

AB010841, respectively. The 16S rRNA gene sequences logical and biochemical characterization of the type of the other 20 species of 7 genera were obtained from strains of Achromobacter xylosoxidans, 4 Alcaligenes the DNA Data Bank of Japan (DDBJ), GenBank, and species, and Bordetella bronchiseptica were performed EMBL data base (Table 2). by the methods described previously (25). The CLUSTAL-W software originally described by The assimilation of glucose, L-arabinose, D-mannitol, Thompson et al (23) was used to align the sequences, and N-acetyl-D-glucosamine, maltose, potassium gluconate, the phylogenetic distance was calculated by using the n-capric acid, adipic acid, dl-malic acid, sodium citrate, neighbor joining (NJ) method. The phylogenetic tree and phenylacetate as carbon and energy sources were was drawn by using Tree View software. tested by using API 20NE (bioMerieux). The results Phenotypic characterization. Morphological physio- were read after 24-hr and 48-hr incubations at 30 C. 432 E. YABUUCHI ET AL

Table 3. Characteristics of six type strains of Achromobacter, Alcaligenes, and Bordetella species

T=type strain . (+) Positive reaction after 3 days or more. ND= No datum. °' Reference 24 , 1 Reference 4, " Reference 23, °' Reference 6, " Present study, 11 Reference 14, x) Kerster et al, IJSB 34: 63, 1984. GENUS ACHROMOBACTER, EMENDATION 433

Table 4. DNA-DNA reassociation rates among the six type strains of Achromobacter, Alcaligenes, and Bordetella species

* Each value represents the average of three experiments .

values among Alcaligenes faecalis, Achromobacter Results xylosoxidans, and 3 other Alcaligenes species are less than 94.3%. Mol% GC Content of DNA and Respiratory Quinone System(Table 3) Phenotypic Characteristics The guanine-plus-cytosine content of DNA from The morphological, physiological, and biochemical Alcaligenes ruhlandii EY 3918Twas 67.7±0.1 mol% features of the type strains of Achromobacter xylosoxi- (averageof three experiments).As shown in Table4, the dans, 4 Alcaligenes species, and Bordetella bronchisep- GC contents of type strain DNA of Achromobacter tica are shown in Table 3. All 6 organisms are nonfer- xylosoxidans(23), Alcaligenes denitrificans (6), Alcali- mentative Gram-negative asporogenous rods and are genes ruhlandii, and Alcaligenes piechaudii are 65.0- motile with peritrichous flagella. The combination of 67.7 mol% (14), whereas that of Alcaligenesfaecalis is abilities to produce oxidative acidity from D-arabinose, 55.7 (14). L-arabinose, 3% ethanol, galactose, glucose, glycerol, D- The respiratory quinone systems of Achromobacter ribose, and D-xylose will be useful to differentiate these xylosoxidans(24), Alcaligenes denitrificans (4), Alcali- organisms. Positive urease reaction is characteristic for genesfaecalis (4), and B. bronchiseptica (4) are report- Bordetella bronchiseptica among these peritrichous ed as ubiquinone 8. species. As shown in Table 3, the type strains of Achro- DNA-DNAHybridization (Table 4) mobacter xylosoxidans and Alcaligenes ruhlandii assim- The reassociation rate of Alcaligenes denitrificans ilated 7 similar substrates, including glucose. Although EY 3839' to labeled DNA of Achromobacterxylosoxi- the remaining 3 type strains of Alcaligenes species uti- dans EY 543' was 63.4%. The reassociation rates of lized each of 4 substrates as carbon and energy sources, cold DNA of Alcaligenes ruhlandii EY 3918' to labeled their assimilation profiles were different. DNA of EY 543' and cold DNA of EY 543' to labeled DNA of EY 3918' were 31.7% and 32.5%,respectively. Discussion The type strains of Achromobacter xylosoxidans and Alcaligenes ruhlandii showed 2.3 to 3.7% similarity Because of the results of phylogenetic analyses of values to the type strain of Alcaligenesfaecalis. 16S rRNA nucleotide sequences and a difference of more than 10% in GC content of DNA, it was confirmed 16S rRNAAnalysis that Achromobacter xylosoxidans, a type species of A phylogenetic tree (Fig. 1) reveals that Alcaligenes genus Achromobacter, and Alcaligenes faecalis, a type denitrificans, Alcaligenes ruhlandii, and Alcaligenes species of genus Alcaligenes, belong to the 2 distinct piechaudii are closely related to Achromobacterxylosox- genera Achromobacter Yabuuchi and Yano 1981 and idans, making a clearly distant cluster from Alcaligenes Alcaligenes Castellani and Charmers 1919, respectively. faecalis. The tree indicatesthat genus Achromobacteris Based on the DNA-DNA similarity values among type appropriately a constituent element of family Alcalige- strains of Achromobacter xylosoxidans, 4 Alcaligenes naceae De Ley et al 1986 (6), together with Alcaligenes species, and Bordetella bronchiseptica, Alcaligenes ruh- and Bordetella. landii and Alcaligenes piechaudii are transferred to As shown in Table5, the 16SrRNA homologyvalues genus Achromobacter as two separate species. Thus among Achromobacterxylosoxidans and 3 Alcaligenes Achromobacter ruhlandii (Packer and Vishniac) comb. species, with the exception of Alcaligenesfaecalis, are nov. and Achromobacter piechaudii (Kiredjian et al) more than 98.4% each others. In contrast,the homology comb. nov. are herein proposed. The synonymy of A. 434 E. YABUUCHI ET AL

Fig. 1. Phylogenetic relationship of 22 species of Achromobacter and six other genera. Distances were calculated by the neighbor-joining (NJ) method. The 16S rRNA sequence accession number of type or reference strains of 22 species of seven genera are shown in Table 4. Alcaligenes denitrificans, Alcaligenes ruhlandii, and Alcaligenes piechaudii constitute a closely related cluster with Achromobacter xylosoxidans. This cluster corresponds to the emended genus Achromobacter. Three genera, Achromobacter, Alcaligenes, and Borde- tella, are regarded to constitute the family Alcaligenaceae. xylosoxidans and A. ruhlandii is denied by the DNA Gram-negative, nonsporeforming, straight rods of 0.8 similarity value of 31.7% between the type strains of the to 1.2 by 2.5 to 3.0.tm with rounded ends. Motile with two species. Vandamme et al (28) reevaluated whole-cell sheathed flagella arranged peritrichously. The number of protein patterns, cellular fatty acid profile, and bio- flagella is 1 to 20 per cell. Strictly aerobic and nonfer- chemical characteristics, and considered A. xylosoxidans mentative. Strains of some species are able to grow and A. denitrificans are distinct species. However, anaerobically with nitrate as electron acceptor. They because of 63.4% reassociation rate and results of perform nitrate respiration combined with nitrite and sequence analysis, Alcaligenes denitrificans Ruger and nitrous oxide respiration (29). Two distinct azurins, Tan (18) is transferred to genus Achromobacter and bacterial blue copper protein probably playing a role of ranked down to subspecies of A. xylosoxidans. Thus electron transfer, are isolated from strains of Achro- Achromobacter xylosoxidans subsp. denitrificans (Ruger mobacter xylosoxidans (27). Nitrite reductases from and Tan) comb. nov. is proposed. This proposal auto- the same strains have been reported to contain both type 1 matically creates the type subspecies of species, Cu and type 2 Cu (22). Urease and DNase are negative. Achromobacter xylosoxidans subsp. xylosoxidans. Strains are also recognized as facultatively lithoautotrophic hydrogen-oxidizers (Knallgas bacteria). The Emendation of Description of the Genus Achromobacter enzyme is an NAD-specific, soluble hydrogenase not Yabuuchi and Yano 1981 (26) accompanied by a membrane-bound hydrogenase (Arag- Based on the comparative study of DNA and 16S no and Schlegel). Some species are also known to form rRNA of the type strains of Achromobacter xylosoxidans dimethyl disulfide (DMDS) from both DL-methionine and 4 Alcaligenes species, the following emended and S-methyl-L-cysteine (4). Nitrate is reduced either to description of the genus Achromobacter is proposed. nitrogen gas or to nitrite. GENUS ACHROMOBACTER, EMENDATION 435 436 E. YABUUCHI ET AL

Catalase and oxidase are produced. Nonhalophilic, Descriptionof the TaxaAssigned to Genus Achromobac- nonhemolytic, and nonpigmented either water-soluble or ter Yabuuchiand Yano1981 -insoluble . Acid is produced oxidatively from some Achromobacterruhlandii (Packer and Vishniac1955) carbohydrates in oxidation-fermentation (OF) medium. comb. nov. Flagellar morphology and hydrogen-oxi- The acid production is generally not strong and takes 3 or dizing ability are as described previously (1, 17, 19). more days for a positive result. Urease, DNase, phenyl- Strain ATCC 15947Tare gram-negative asporogenous alanine deaminase, lysine and ornithine decarboxylase, rod. Nonfermentative,nonhemolytic, and nonhalophilic. arginine dihydrolase, and gelatinase are negative. The Able to grow on MacConkeyagar, at the presenceof 5% assimilation of dl-malic acid, adipic acid, and pheny- sodium chloride,and at 41 C. Catalaseand oxidase pos- lacetate is uniformly positive in 3 species, whereas L-ara- itive. Alkalinize Simmons' citrate medium within 2 binose, D-mannitol, D-mannose, maltose, and N-acetyl-D- days and malonate broth after 3 days of incubation. glucosamine are uniformly negative in 3 species. The Reduce nitrateto nitrite. Does not produce gas in nitrate assimilation is negative for sodium citrate by A. piechaudii, broth. Oxidative acidity is produced in OF medium potassium gluconate by A. xylosoxidans subsp. denitrif- base from D-arabinosewithin 48 hr, from L-arabinose, icans, and n-capric acid is negative by both organisms. glucose, D-ribose,and D-xyloseafter 72-hr incubation. Although soil and water are regarded as natural habi- Acid was not produced from fructose, galactose, cel- tats, little ecological information is available. Sometimes lobiose, lactose, maltose, melibiose, sucrose, trehalose, isolated from a hospital environment and from human raffinose, melezitose,glycerol, adonitol, dulcitol, inosi- clinical specimens with pathological significance or just tol, mannitol,sorbitol, salicine, and inulin. Acylamidase as a contaminant. The mol% GC of the DNA is 65-67.7. test becomespositive after 3 days of incubation. Urease, Genus Achromobacter is regarded as a member of phenylalanine deaminase, and DNase are negative. the family Alcaligenaceae De Ley et al 1986 (6), belong- Esculin,gelatin, starch, and Tween80 are not hydrolyzed. ing to the Proteobacteria beta group (21). The present Lysine and ornithine decarboxylase and arginine dihy- members of genus Achromobacter are A. xylosoxidans, drolase are negative. Neither water-soluble nor water- A. xylosoxidans subsp. denitrificans, A. ruhlandii, and A. insoluble pigments are produced. Assimilates glucose, piechaudii. Type species: Achromobacter xylosoxidans adipic acid, capric acid, dl-malic acid, citrate gluconate, Yabuuchi and Yano 1981 477. and phenyl acetate. The assimilationprofile 'of 12 organic substrates is similar to that of A. xylosoxidans. The Emended Description of Achromobacter xylosoxidans natural habitat is known so far to be soil (16). (ex Yabuuchi and Ohyama 1971) Yabuuchi and Yano The mol% GC of DNA is 67.7. The DNA-DNA 1981477 similarityvalue between the type strains of this species The morphological and physiological characteristics and Achromobacterxylosoxidans is 32.5%. are as described for the genus. Colonies on heart infusion Type strain is ATCC 15947. agar are 1 mm in diameter, low convex with entire mar- Achromobacter piechaudii (Kiredjian et al 1986) gin, moist, and with a glistening surface. Grayish-white, comb. nov. Morphological,physiological, biochemical semitranslucent. Able to grow anaerobically in the pres- characteristics,cellular protein electrophoresispatterns, ence of nitrate or nitrite by denitrification. Among 12 and mol% GC of DNA from the type strain are as substrates incorporated in API 20NE, glucose, gluconate, reported previously (14). Acid was not produced in citrate, adipic acid, capric acid, dl-malic acid, and phenyl- OF medium base supplementedwith each of 25 carbo- acetate are assimilated as carbon and energy sources. hydratesincluding glucose. Among the 12 organic sub- The assimilation profile of 12 organic substrates is sim- strates,dl-malic acid, adipic acid, gluconate,and phenyl- ilar to that of A. ruhlandii. Phenotypic characteristics, acetate are assimilated. Glucose and citrate are not uti- cellular fatty acid composition, antibiotic susceptibility, lized. The mol% GC of DNA from 7 strains is 64-65 source of isolation including human clinical specimens, (14). unsterilized "distilled water," and chlorhexidine solutions Type strain is ATCC 15173. are as described previously (23, 24). Achromobacter xylosoxidans subsp. denitrificans Native dissimilatory nitrite reductases contain type 1 (Roger and Tan 1983) comb. nov. Morphological, Cu (blue copper) and type 2 Cu (nonblue copper) (22). physiological, and biochemical characteristics are as Facultatively lithoautotrophic hydrogen-oxidizer (Knall- described previously (18). Able to grow at 41 C. Acid gas bacteria). The respiratory quinone system is is producedin OF mediumfrom 3% ethanolwithin 48 hr ubiquinone 8. and from glycerol after 72 hr. Among the 12 organic The type strain is ATCC 27061. The mol% GC of substrates,dl-malic acid,adipic acid, citrate, and phenyl- DNA ranges from 66.9 to 69.8 (23). acetate are assimilated. The respiratoryquinone system GENUS ACHROMOBACTER, EMENDATION 437 is ubiquinone 8 (4). The mol% GC of DNA from the tion in microdilutionwells as an alternative to membrane fil- type strain is 68.5 (6). The DNA-DNA similarity value ter hybridization in which radioisotopes are used to determine between type strains of the species to Achromobacter genetic relatedness among bacterial strains. Int. J. Syst. xylosoxidans is 63.4%, subspecies level similarity. B acteriol. 39: 224-229. 9) Ezaki, T., Saidi, S.M., Liu, S.-L., Hashimoto, Y., Yama- Type strain is ATCC 15173. moto, H., and Yabuuchi,E. 1990.Rapid procedureto determine Achromobacter xylosoxidans subsp. xylosoxidans the DNA base composition from small amounts of gram-pos- (Yabuuchi and Yano 1981) comb. nov. Type subspecies itive bacteria. FEMS Microbiol. Lett. 67: 127-130. of A. xylosoxidans. Description of the subspecies corre- 10) Hendrie, M.S., Holding, A.J., and Shewan, J.M. 1974. sponds with those of species A. xylosoxidans. Type strain Emended descriptions of the genus Alcaligenes and of Alcali- for the subspecies is ATCC 27061, same with species A. genes faecalis and proposal that the generic name Achro- xylosoxidans. mobacter be rejected: status of the named species of Alcali- genes and Achromobacter. Request for an Opinion. Int. J. Syst. Bacteriol. 24: 534-550. A part of this work was supportedby the Grantin Aid in 1997 11) Holt, J.G. 1979. International Committee on Systematic to E.Y.from the Ministryof Healthand Welfare,Japan. Bacteriology, Judicial Commission, Minutes of the Meeting, One of the authors(E.Y.) is gratefulto Professor Shinnichiro 3 September 1978, Minute 24, Rejection of the genus name Suzuki,Graduate School of Science,Osaka University,for his Achromobacter. Int. J. Syst. Bacteriol. 29: 267-269. informationon nitrite reductaseand critical readingof the man- 12) Kawamura, Y., Hou, X.G., Todome, Y., Sultana, F., Hirose, uscript. K., Shu, S., Ezaki, T., and Ohkuni, H. 1998. Streptococcus peroris sp. nov. and Streptococcus infantis sp. nov.; new References members of the Streptococcus mitis group, isolated from human clinical specimens. Int. J. Syst. Bacteriol. 48: in 1) Aragno,M., and Schlegel,H.G. 1977.Alcaligenes ruhlandii press. (Packerand Vishniac)comb. nov., a peritrichoushydrogen 13) Kersters, K., and De Ley, J. 1984. Genus Alcaligenes Castel- bacteriumpreviously assigned to Pseudomonas.Alcaligenes lani and Chalmers 1919, 936', p. 365. In Krieg, N.R., and ruhlandii(Packer and Vishniac1955) Aragno and Schlegel Holt, V. (eds), Bergey's manual of systematic bacteriology, 1977AL. Int. J. Syst. Bacteriol.27: 279-281. Vol. 1, Williams & Wilkins, Baltimore/London. 2) Aragno,M., and Schlegel,H.G. 1992. The mesophilichydro- 14) Kiredjian, M., Holmes, B., Kersters, K., Guilvout, I., and De gen-oxidizing(knallgas) bacteria, p. 344-384.In Balows, A., Ley, J. 1986. 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