Of Moraxella Osloensis

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Of Moraxella Osloensis INTERNATIONAL JOURNAL OF SYSTEMATICBACTERIOLOGY, July 1993, p. 474-481 Vol. 43, No. 3 0020-7713/93/030474-08$02.00/0 Copyright 0 1993, International Union of Microbiological Societies Moracxella ZincoZnii sp. nov., Isolated from the Human Respiratory Tract, and Reevaluation of the Taxonomic Position of Moraxella osloensis P. VANDAMME,'.,* M. GILLIS,l M. VANCA"EYT,l B. HOSTE,' K. KERSTERS,' AND E. FALSEN3 Laboratorium voor Microbiologie, University of Ghent, K L. Ledeganchtraat 35, B-9000 Ghent, and Department of Microbiology, University Hospital, Antwerp, Belgium, and Culture Collection, Department of Clinical Bacteriology, University of Gotebolg, S-413 46 Gotebolg, Sweden3 A polyphasic taxonomic study was performed to determine the relationships of 10 Muraellu-like strains isolated mainly from the human respiratory tract in Sweden. Two of the strains formed a separate subgroup on the basis of both their protein contents and their fatty acid contents. However, the overall protein and fatty acid profiles revealed that all 10 strains were highly related. Representative strains of the two subgroups exhibited high DNA binding values (98%) with each other and had an identical DNA base ratio (44 mol% G+C), DNA-rRNA hybridizations revealed that this taxon can be included in the genus Muraxelh, which is only distantly related to phenotypically similar genera, such as the genera Neisseriu and Kingelh, The results of an extensive phenotypic analysis indicated that the general biochemical profile of the 10 strains conforms with the description of the genus Muraxelh given in Bergey 3 Manual of Systematic Bacteriology, We therefore consider these organisms members of a new Muraxelh species, for which the name Moraxellu lincolnii is proposed, Furthermore, we also conclude that MumelZu osloensis belongs, genotypically as well as phenotyp- ically, to the genus Muraxelh. Between 1980 and 1989, 10 Moraxella-like strains were of special peptone (catalog no. L72; Oxoid), 5 g of Lab isolated in Sweden, mainly from the human respiratory tract. Lemco powder (catalog no. L29; Oxoid), 5 g of yeast extract In October 1980, immunotyping (Ouchterlony immunodiffu- (catalog no. L21; Oxoid), 5 g of sodium chloride, 2 g of sion and immunoelectrophoresis) was performed on the first sodium succinate hexahydrate, 2 g of sodium L-glutamate three isolates, and the results revealed that the strains monohydrate, 1 g of magnesium chloride hexahydrate, and represented a distinct taxon which was provisionally named 15 g of agar no. 3 (catalog no. L13; Oxoid); the final pH is "Moraxella lincolnii." Subsequent isolates were rapidly 7.0. This medium was supplemented with 5% (vol/vol) horse identified. In the present study we used a polyphasic ap- blood and is referred to below as standard medium. Optimal proach to determine the phenotypic and genotypic affiliation growth results were obtained at 28°C in a microaerobic of these organisms. We found that the 10 strains constitute a atmosphere containing 5% O,, 10% CO,, and 85% N,. M. new species belonging to the genus Moraxella, and we osloensis strains and reference strains belonging to other propose the name Moraxella Zincolnii sp. nov. This name is Moraxella species were cultivated aerobically at 28°C on used below. In view of the new data and recently published heart infusion agar (Difco). results (9), we also reevaluated the taxonomic position of Phenotypic tests. Phenotypic tests were performed on all Moraxella osloensis as the arguments used by Rossau et al. M. lincolnii strains and on Moraxella reference strains. The to remove this species from the genus Moraxella (18) were morphology of cells was evaluated by Gram staining. Motil- not conclusive. ity was observed in young cultures by examining wet mounts in broth by phase-contrast microscopy. The following tests were performed as described previously (5): presence of MATERIALS AND METHODS oxidase, catalase, p-galactosidase, DNase, and urease; pro- duction of indole; reduction of nitrate and nitrite; denitrifi- Bacterial strains and growth conditions. All of the strains cation; growth under anaerobic conditions; growth on Dri- used are listed in Table 1. We obtained weak growth of M. galski agar; growth at different temperatures; growth in the Lincolnii strains on several blood agar bases, including 5% presence of different concentrations of NaCl; oxidation or (vol/vol) horse blood in Mueller-Hinton agar (catalog no. fermentation of D-glucose; liquefaction of gelatin; proteoly- CM337; Oxoid, Ltd., Basingstoke, United Kingdom), nutri- sis on Loffler slants; hydrolysis of Tween 80; tolerance of ent agar no. 2 (catalog no. CM67; Oxoid), Columbia agar penicillin (lO-Fg discs); hemolysis and odor on horse blood (catalog no. CM331; Oxoid), and heart infusion agar (catalog agar; and formation of pigment on nutrient agar. Acid no. 0038-01-5; Difco Laboratories, Detroit, Mich.). Growth production from D-glucose, maltose, D-fructose, and sucrose was not markedly enhanced on chocolate agar plates. The was evaluated in a medium containing 15 g of Proteose best growth results were obtained on a medium routinely Peptone no. 3 (catalog no. 0122; Difco), 4.5 g of NaCl, 3.0 g used in our department for the cultivation of fastidious of Na,HPO, . 2H,O, 1.5 g of sodium alginate, and 17 g of Campylobacter strains. This medium contains (per liter) 10 g Bacto Agar (catalog no. 0140; Difco) in 1,000 ml of placenta extract (the pH was adjusted to 7.7). This basal medium was supplemented with 2.5 ml of phenol red (0.5% solution), 5 ml * Corresponding author. of hemin (0.2% solution), 90 ml of ascitic fluid, and 10 ml of 474 VOL.43, 1993 MORAXELLA LINCOLNII SP. NOV. 475 TABLE 1. Strains used Received Source, if known Taxon Strain" from": M. lincolnii CCUG 9405' (= LMG 5127') Lincoln Human nasopharynx, 6-mo-old female with fever (1980, Sweden) M. lincolnii CCUG 9406 (= LMG 10487) Lincoln Human throat, 1.5-yr-old male, respiratory tract infection (1980, Sweden) M. lincolnii CCUG 9511 (= LMG 10488) Lange Human nasopharynx, 1.5-yr-old female (1980, Sweden) M. lincolnii CCUG 11973 (= LMG 10489) Dennford Human nasopharynx, 2-yr-old child, whooping cough specimen (1982, Sweden) M. lincolnii CCUG 17467 (= LMG 10490) PHL Human nasopharynx, 21-yr-old female (1985, Sweden) M. lincolnii CCUG 18789 (= LMG 10491) Dennford Human eye, 5-mo-old male (1986, Sweden) M. lincolnii CCUG 20072 (= LMG 10492) Bronnestam Human respiratory tract, 11-mo-old male (1987, Sweden) M. Iincolnii CCUG 20073 (= LMG 10493) Bronnestam Human nasopharynx, 1-yr-old male (1987, Sweden) M. lincolnii CCUG 21674 (= LMG 10494) PHL Human nasopharynx, 6.5-yr-old female (1987, Sweden) M. lincolnii CCUG 24769 (= LMG 10495) PHL Human blood, bursitis of the left knee (1989, Sweden) M. osloensis LMG 987 (= ATCC 10973) TRS M. osloensis LMG 1043 (= ATCC 19961 = TRS Human urine (1951, Norway) NCTC 10756) M. osloensis LMG 1044 (= ATCC 19963 = TRS Human nose (1951, Norway) NCTC 10749) M. osloensis CCUG 63 (= LMG 6916) Lincoln Human urine, 10-yr-old girl, suspected pyelonephritis (1968, Sweden) M. osloensis CCUG 350T (= LMG 5131T = B~vre Human cerebrospinal fluid ATCC 19976') M. osloensis CCUG 11977 (= LMG 6917) PHL Human blood, 82-yr-old male with pneumonia (1982, Sweden) M. osloensis CCUG 13305 (= LMG 6918) PHL Human abdomen, 15-yr-old female, pus from appendix (1983, Sweden) M. osloensis CCUG 13369 (= LMG 6919) PHL Human blood, 8-yr-old male (1983, Sweden) M. osloensis CCUG 13705 (= LMG 6920) PHL Human cerebrospinal fluid, 24-yr-old female, meningitis (1983, Sweden) M. atlantae CCUG 6415T (= LMG 5133= = Bgvre Human blood (United States) ATCC 29525') M. bovis CCUG 2133T (= LMG 986T = Bmre Cattle, pinkeye ATCC lO9OOT) M. canis CCUG 8415AT (= LMG 11194T) PHL Human wound, 29-yr-old woman, dog bite (1979, Sweden) M. catarrhalis CCUG 353T (= LMG 11192T = Bgvre ATCC 25238T) M. caviae CCUG 35ST (= LMG 5129T = B0vre Pharynx, guinea pig (United States) ATCC 14659T = CCUG 2132T) M. cuniculi CCUG 2154' (= LMG 8382T = Berger Rabbit oral mucosa ATCC 14688') M. lacunata CCUG 4441* (= LMG 5301T = CIP Subacute conjunctivitis CIP A182T) M. lacunata LMG 1009 (= ATCC 17952 = TRS CCUG 4862 = NCTC 7919 M. nonlique- CCUG 348T (= ATCC 19975 = Bmre Nose swab faciens NCTC 10464T) M. nonlique- LMG 1011 (= ATCC 17954) TRS Human throat swab (1950, Canada) faciens M. ovis CCUG 354' (= LMG 8381T = Bgvre mine eye, conjunctivitis ATCC 33078T) ~ ~ ~ ~~ ATCC, American weCulture Collection, Rockville, Md.; Berger, U. Berger, Heidelberg, Germany; Bronnestam, R. Bronnestam, Skovde, Sweden; B0vre, K. Mwe, Oslo, Norway; CCUG, Culture Collection, University of Goteborg, Goteborg, Sweden; CIP, Collection de I'Institut Pasteur, Paris, France; Dennford, J. Dennford, Bor As, Sweden; Lange, S. Lange, Gijteborg, Sweden; LMG, Culture Collection of the Laboratorium voor Microbiologie, University of Ghent, Ghent, Belgium; Lincoln, K. Lincoln, Goteborg, Sweden; NCTC, National Collection of Type Cultures, London, United Kingdom; PHL, Public Health Laboratory Service, Giiteborg, Sweden; TRS, Torry Research Station, Aberdeen, Scotland. a 30% carbohydrate solution. API ZYM tests were per- amidase, lysine arylamidase, proline arylamidase, cr-glut- formed according to the recommendations of the manufac- amyl-a-glutamic acid arylamidase, glycyl-phenylalanine turer (MI bioM&rieux, La Balme-les-Grottes, Montalieu- arylamidase, prolyl-arginine arylamidase, seryl-methionine Vercieu, France). arylamidase, and alanyl-phenylalanyl-prolyl-alaninearyla- The following enzymes belonging to a noncommercial
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