DNA-DNA Hybridization Data Research Articles

Aeromonas aquariorum sp. nov., isolated from aquaria of ornamental fish

During a survey to determine the prevalence of Aeromonas strains in water and skin of imported ornamental fish, 48 strains presumptively identified as Aeromonas were isolated but they could not be identified as members of any previously described Aeromonas species. These strains were subjected to a polyphasic approach including phylogenetic analysis derived from gyrB, rpoD and 16S rRNA gene sequencing, DNA-DNA hybridization, MALDI-TOF MS analysis, genotyping by RAPD and extensive biochemical and antibiotic susceptibility tests in order to determine their taxonomic position. Based on the results of the phylogenetic analyses and DNA-DNA hybridization data, we describe a novel species of the genus Aeromonas, for which the name Aeromonas aquariorum sp. nov. is proposed, with strain MDC47T (=DSM 18362T =CECT 7289T) as the type strain. This is the first Aeromonas species description based on isolations from ornamental fish.

Aeromicrobium ponti sp. nov., isolated from seawater

A novel actinomycete strain, designated HSW-1T, was isolated from seawater collected from Hwasun Beach on the coast of Jeju Island, Republic of Korea. The cells were aerobic, Gram-positive, oxidase-negative, catalase-positive, non-motile rods. The colonies were circular, smooth, convex and yellow in colour. The cell-wall peptidoglycan of this organism contained ll-diaminopimelic acid as the diagnostic diamino acid. The polar lipids included phosphatidylinositol, diphosphatidylglycerol, phosphatidylglycerol and three unknown phospholipids. The predominant menaquinone was MK-9(H4). The major fatty acids were C18 : 1 omega 9c, C16 : 0, C16 : 0 2-OH and 10-methyl C18 : 0. The DNA G+C content was 74.0 mol%. A neighbour-joining tree based on 16S rRNA gene sequences showed that this organism falls within the radiation of the genus Aeromicrobium. The closest phylogenetic neighbours were the type strains of Aeromicrobium erythreum (98.2 % 16S rRNA gene sequence similarity), Aeromicrobium alkaliterrae (97.9 %), Aeromicrobium marinum (97.3 %) and Aeromicrobium fastidiosum (97.0 %). The DNA-DNA relatedness values between the novel isolate and its closest relative, A. erythreum DSM 8599T, ranged between 32 and 36 %. On the basis of the phenotypic and DNA-DNA hybridization data, strain HSW-1T represents a novel species of the genus Aeromicrobium, for which the name Aeromicrobium ponti sp. nov. is proposed. The type strain is HSW-1T (=DSM 19178T=KACC 20565T).

Nocardiopsis quinghaiensis sp. nov., isolated from saline soil in China

A previously unknown Gram-positive, obligately aerobic actinomycete, YIM 28A4(T), was isolated from a sample of saline soil collected from the Qaidam Basin in Qinghai Province, north-west China, and was investigated using a polyphasic taxonomic approach. The strain grew well on most of the media tested, producing white to pale-yellow substrate mycelium, white aerial mycelium and straight to flexuous hyphae. The substrate mycelium was well developed and fragmented with age; the aerial mycelium produced long, straight spore chains. The spore chains were composed of non-motile, smooth-surfaced, rod-shaped spores. No diffusible pigments were produced on any of the media tested. The strain grew in the presence of 0-10 % (w/v) NaCl and at pH 6.0-8.0, with optimum growth occurring at 3 % (w/v) NaCl and pH 7.0. It grew at 10-37 degrees C, the optimum growth temperature being 28 degrees C. Whole-cell hydrolysates of strain YIM 28A4(T) contained meso-diaminopimelic acid and no diagnostic sugars. The predominant phospholipids were phosphatidylcholine, phosphatidylglycerol and diphosphatidylglycerol. The predominant menaquinones were MK-10, MK-10(H(2)), MK-11 and MK-11(H(2)). The major cellular fatty acids were iso-C(16 : 0), anteiso-C(15 : 0) and anteiso-C(17 : 0). The DNA G+C content was 67.1 mol%. The morphological and chemotaxonomic characteristics of the isolate matched those described for Nocardiopsis species. A phylogenetic analysis based on 16S rRNA gene sequence comparisons confirmed that strain YIM 28A4(T) was a member of the genus Nocardiopsis and most closely related to the type strains Nocardiopsis aegyptia DSM 44442(T) and Nocardiopsis halotolerans DSM 44410(T), showing 98.1 and 97.8 % 16S rRNA gene sequence similarity, respectively. Strain YIM 28A4(T) can be differentiated from these type strains by using phenotypic, phylogenetic and DNA-DNA hybridization data. On the basis of the polyphasic evidence, strain YIM 28A4(T) represents a novel species of the genus Nocardiopsis, for which the name Nocardiopsis quinghaiensis sp. nov. is proposed. The type strain is YIM 28A4(T) (=DSM 44739(T) =CGMCC 4.3494(T)).

Virgibacillus kekensis sp. nov., a moderately halophilic bacterium isolated from a salt lake in China

A Gram-positive, moderately halophilic, motile, strictly aerobic, endospore-forming, oxidase- and catalase-positive, rod-shaped bacterium, strain YIM kkny16(T), was isolated from a saline mud sample collected from the Keke salt lake in the Qaidam Basin, north-west China. This isolate grew in the presence of 0-25 % (w/v) NaCl and at pH 6.0-10.0 and 10-50 degrees C; optimum growth was observed with 10 % (w/v) NaCl and at pH 7.0 and 37 degrees C. Strain YIM kkny16(T) had meso-diaminopimelic acid as the diagnostic diamino acid, MK-7 as the predominant respiratory quinone, with a significant amount of MK-6, and anteiso-C(15 : 0), iso-C(14 : 0) and C(16 : 1)omega7c alcohol as major fatty acids. Major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The DNA G+C content was 41.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences confirmed that strain YIM kkny16(T) was a member of the genus Virgibacillus, exhibiting sequence similarities of 94.9-97.3 % to the type strains of recognized Virgibacillus species. Strain YIM kkny16(T) could be differentiated from recognized Virgibacillus species based on phenotypic characteristics, chemotaxonomic differences, phylogenetic analysis and DNA-DNA hybridization data. On the basis of evidence from this polyphasic study, strain YIM kkny16(T) is considered to represent a novel species of the genus Virgibacillus, for which the name Virgibacillus kekensis sp. nov. is proposed. The type strain is YIM kkny16(T) (=DSM 17056(T)=CGMCC 1.6298(T)).

Classification of 'Streptomyces tenebrarius' Higgins and Kastner as Streptoalloteichus tenebrarius nom. rev., comb. nov., and emended description of the genus Streptoalloteichus

Phylogenetic analysis of 'Streptomyces tenebrarius' NBRC 16177 on the basis of the 16S rRNA gene sequence revealed that this organism was related to Streptoalloteichus hindustanus. It possessed glutamic acid, alanine and meso-diaminopimelic acid as cell-wall amino acids, galactose, mannose and glucose as whole-cell sugars and the menaquinones MK-10(H(6)), MK-10(H(4)), MK-9(H(6)) and MK-9(H(4)). The chemotaxonomic characteristics of this strain were consistent with those of the genus Streptoalloteichus. DNA-DNA hybridization, physiological, biochemical and chemotaxonomic data revealed that 'Streptomyces tenebrarius' can be easily differentiated from the other member of the genus Streptoalloteichus and that it merits separate species status. The phenotypic and genetic evidence reveals that 'Streptomyces tenebrarius' represents a novel species of the genus Streptoalloteichus; the name Streptoalloteichus tenebrarius (ex Higgins and Kastner) nom. rev., comb. nov. is proposed for this species. The type strain is NBRC 16177(T) (=ATCC 17920(T) =DSM 40477(T) =JCM 4838(T) =NRRL B-12390(T) =ISP 5477(T)).

Classification of halo(alkali)philic and halo(alkali)tolerant methanotrophs provisionally assigned to the genera Methylomicrobium and Methylobacter and emended description of the genus Methylomicrobium

The taxonomic positions of four aerobic, obligately halo(alkali)philic/-tolerant, methanotrophic bacteria previously affiliated with the genera Methylobacter ('Methylobacter alcaliphilus' strains 20Z and 5Z) and Methylomicrobium (Methylomicrobium strains AMO1 and NI) were investigated. Phylogenetic analysis of 16S rRNA gene sequences indicated that the strains form a separate branch within the type I methanotrophic bacteria and are closely related to Methylomicrobium pelagicum. DNA-DNA hybridization data revealed relatively low levels of relatedness of Methylomicrobium sp. AMO1 and Methylomicrobium sp. N1 with each other and with previously described species of the genera Methylomicrobium and Methylobacter (<55 %), indicating that they may represent novel species. Based on the results presented here and on previously reported morphological and physiological characteristics, we classify these halotolerant and halophilic methanotrophic strains as representing novel species within the genus Methylomicrobium: Methylomicrobium alcaliphilum sp. nov. (type strain 20Z(T) =VKM B-2133(T) =NCIMB 14124(T); reference strain 5Z =VKM B-2180), Methylomicrobium japanense sp. nov. (type strain NI(T) =VKM B-2462(T) =FERM BP-5633(T) =NBRC 103677(T)) and Methylomicrobium kenyense sp. nov. (type strain AMO1(T) =NCCB 97157(T) =NCIMB 13566(T) =VKM B-2464(T)). The genus Methylomicrobium has been emended in its description.

Lysobacter capsici sp. nov., with antimicrobial activity, isolated from the rhizosphere of pepper, and emended description of the genus Lysobacter

The taxonomic position of a novel bacterial strain, YC5194(T), with antimicrobial activity, isolated from the rhizosphere of pepper in Jinju, South Korea, was studied using a polyphasic approach. Cells of the strain were Gram-negative, rod-shaped, facultative anaerobes. It grew at a temperature of 15-37 degrees C (optimum 28 degrees C). Growth of the strain occurred between pH 5.5 and 8.5, with an optimum of pH 7.0-7.5. The strain inhibited mycelial growth of Pythium ultimum, Colletotrichum gloeosporioides, Fusarium oxysporum, Botrytis cinerea, Rhizoctonia solani and Botryosphaeria dothidea and growth of Bacillus subtilis. The G+C content of the total DNA was 65.4 mol%. The 16S rRNA gene sequence of the strain was most closely related to species of the genus Lysobacter (<94.0 to >99.0 % sequence similarity). Chemotaxonomic data (major quinone, Q-8; major polar lipids, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and phosphatidyl-N-methylethanolamine; major fatty acids, iso-C(15 : 0), summed feature 3, C(16 : 0), iso-C(17 : 1)omega9c and C(18 : 1)omega7c) supported the affiliation of strain YC5194(T) to the genus Lysobacter. Phylogenetic analysis based on 16S rRNA gene sequences, DNA-DNA hybridization data and biochemical and physiological characteristics strongly supported the genotypic and phenotypic differentiation of strain YC5194(T) from species of Lysobacter with validly published names. Strain YC5194(T) therefore represents a novel species, for which the name Lysobacter capsici sp. nov. is proposed. The type strain is YC5194(T) (=KCTC 22007(T) =DSM 19286(T)).

Lysobacter spongiicola sp. nov., isolated from a deep-sea sponge

An aerobic, Gram-negative bacterium, strain KMM 329(T), was isolated from a deep-sea sponge specimen from the Philippine Sea and subjected to a polyphasic taxonomic investigation. Comparative 16S rRNA gene sequence analysis showed that strain KMM 329(T) clustered with the species of the genus Lysobacter. The highest level of 16S rRNA gene sequence similarity (97.0 %) was found with respect to Lysobacter concretionis KCTC 12205(T); lower values (96.4-95.2 %) were obtained with respect to the other recognized Lysobacter species. The value for DNA-DNA relatedness between strain KMM 329(T) and L. concretionis KCTC 12205(T) was 47 %. Branched fatty acids 16 : 0 iso, 15 : 0 iso, 11 : 0 iso 3-OH and 17 : 1 iso were found to be predominant. Strain KMM 329(T) had a DNA G+C content of 69.0 mol%. On the basis of the phenotypic, chemotaxonomic, DNA-DNA hybridization and phylogenetic data, strain KMM 329(T) represents a novel species of the genus Lysobacter, for which the name Lysobacter spongiicola sp. nov. is proposed. The type strain is KMM 329(T) (=NRIC 0728(T) =JCM 14760(T)).

Paracoccus marinus sp. nov., an adonixanthin diglucoside-producing bacterium isolated from coastal seawater in Tokyo Bay

Two novel marine, Gram-negative, non-motile, catalase- and oxidase-positive, aerobic bacteria were isolated from coastal seawater in Tokyo Bay. Analysis of almost-complete 16S rRNA gene sequences showed that the two isolates are members of the genus Paracoccus, sharing highest 16S rRNA gene sequence similarity (96.5 %) with Paracoccus aminophilus NBRC 16710(T). The DNA-DNA reassociation values between P. aminophilus NBRC 16710(T) and these isolates were only 10-20 %, in contrast to the high DNA relatedness between the two isolates (89 %). At least 1 % (w/v) NaCl was required for growth. Cellular fatty acid profiles revealed C(18 : 1)omega7c as the major component and C(10 : 0) 3-OH as the major hydroxy fatty acid. Ubiquinone-10 was detected as the major respiratory quinone. The G+C content of the genomic DNA of both strains was 69 mol%. On the basis of DNA-DNA hybridization data and physiological and chemotaxonomic characteristics, it is proposed that these strains should be placed in a novel species, Paracoccus marinus sp. nov. The type strain is KKL-A5(T) (=NBRC 100637(T) =CIP 108500(T)); KKL-B9 (=NBRC 100640) is a reference strain.

Arthrobacter oryzae sp. nov. and Arthrobacter humicola sp. nov.

Two novel bacterial strains were isolated from a paddy soil sample collected in Japan using GPM agar plates supplemented with superoxide dismutase and/or catalase. The strains were Gram-positive, catalase-positive and motile, with lysine as the peptidoglycan diagnostic diamino acid and acetyl as the peptidoglycan acyl type. The major menaquinone was MK-9(H(2)). Mycolic acids were not detected. The G+C content of the DNA was 66-68 mol%. On the basis of phenotypic analysis, 16S rRNA gene sequence comparisons and DNA-DNA hybridization data, it is proposed that these strains represent two novel species, Arthrobacter oryzae sp. nov. (type strain is KV-651(T)=NRRL B-24478(T)=NBRC 102055(T)) and Arthrobacter humicola sp. nov. (type strain is KV-653(T)=NRRL B-24479(T)=NBRC 102056(T)), respectively.

Salinicoccus salitudinis sp. nov., a new moderately halophilic bacterium isolated from a saline soil sample

A novel pale-yellow-pigmented, moderately halophilic, facultatively alkaliphilic, non-motile, non-spore-forming, catalase- and oxidase-positive, obligately aerobic Gram-positive coccus, strain YIM-C678T was isolated from a saline soil sample collected from a hypersaline habitat in the Qaidam basin, northwest China. The organism grew at 4-37 degrees C and pH 6.0-11.0, with optimum growth at 25 degrees C and pH 8.0. Strain YIM-C678T grew optimally in the presence of 10-12% (w/v) NaCl and growth was observed in 1-25% (w/v) NaCl. The cell wall murein type was L: -Lys-Gly5. Major cellular fatty acids were anteiso-C15:0, iso-C15:0, iso-C16:0 and C16:0. Menaquinone 6 (MK-6) was the major respiratory quinone. The DNA G + C content was 46.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strain YIM-C678T belonged to the family Staphylococcaceae and was most closely related to the eight described species of the genus Salinicoccus with sequence similarities from 92.2 (S. luteus YIM 70202T) to 97.5% (S. kunmingensis YIM Y15T). The DNA-DNA relatedness between strain YIM-C678T and S. kunmingensis YIM Y15T was 35.4%. Chemotaxonomic data and 16S rRNA gene sequence analysis supported the affiliation of strain YIM-C678T with the genus Salinicoccus. The combination of phylogenetic analysis, phenotypic characteristics, chemotaxonomic differences and DNA-DNA hybridization data supported the view that the bacterium represents a novel species of the genus Salinicoccus, for which the name Salinicoccus salitudinis sp. nov. is proposed, with YIM-C678T (=DSM 17846 = CGMCC 1.6299) as the type strain.

Reduction of chromate, selenite, tellurite, and iron (III) by the moderately thermophilic bacterium Bacillus thermoamylovorans SKC1

A moderately thermophilic, facultatively anaerobic bacterium capable of reducing Cr(VI) (strain SKC1) was isolated from municipal sewage. Based on the analysis of the 16S rRNA gene nucleotide sequence and DNA-DNA hybridization data, strain SKC1 was identified as a representative of the species Bacillus thermoamylovorans. B. thermoamylovorans SKC1 is capable of reducing chromate with L-arabinose as an electron donor with an optimum at 50 degrees C and neutral pH. The culture is able to reduce Cr(VI) at its initial concentration in the medium of up to 150 mg/l. In addition to chromate, strain SKC1 is capable of reducing selenite and tellurite, as well as soluble forms of Fe(III). It was shown that Cr(VI), Te(IV), and Se(IV) exert a bacteriostatic effect on strain SKC1, and the reduction of these anions performs the detoxification function. This is the first communication on the reduction of chromate, selenite, tellurite, and soluble Fe(III) species by a culture of thermophilic bacilli.

Taxonomic recommendations for British birds: Fourth report

Taxonomic recommendations for British birds: Fourth report

Topology testing of phylogenies using least squares methods

BackgroundThe least squares (LS) method for constructing confidence sets of trees is closely related to LS tree building methods, in which the goodness of fit of the distances measured on the tree (patristic distances) to the observed distances between taxa is the criterion used for selecting the best topology. The generalized LS (GLS) method for topology testing is often frustrated by the computational difficulties in calculating the covariance matrix and its inverse, which in practice requires approximations. The weighted LS (WLS) allows for a more efficient albeit approximate calculation of the test statistic by ignoring the covariances between the distances.ResultsThe goal of this paper is to assess the applicability of the LS approach for constructing confidence sets of trees. We show that the approximations inherent to the WLS method did not affect negatively the accuracy and reliability of the test both in the analysis of biological sequences and DNA-DNA hybridization data (for which character-based testing methods cannot be used). On the other hand, we report several problems for the GLS method, at least for the available implementation. For many data sets of biological sequences, the GLS statistic could not be calculated. For some data sets for which it could, the GLS method included all the possible trees in the confidence set despite a strong phylogenetic signal in the data. Finally, contrary to WLS, for simulated sequences GLS showed undercoverage (frequent non-inclusion of the true tree in the confidence set).ConclusionThe WLS method provides a computationally efficient approximation to the GLS useful especially in exploratory analyses of confidence sets of trees, when assessing the phylogenetic signal in the data, and when other methods are not available.

Fluviols, bicyclic nitrogen-rich antibiotics produced by Pseudomonas fluorescens.

Fifteen fluorescent pseudomonads, isolated from the rhizosphere of agricultural plants, were similar in both their phenotypic properties and the chemical nature of produced pigments, to the previously described Pseudomonas fluorescens var. pseudoiodinum. DNA-DNA hybridisation data showed their genetic similarity (but not identity) to different biovars of P. fluorescens. A family of antibiotics-fluviols belonging to pyrazolo-[4,3-e]as-triazine derivatives was isolated from studied strains; isolation, properties, antimicrobial and antitumour activity of fluviols are described.

Microorganisms Involved in the Fermentation Process of Panettone Sourdough Maintaining in Japan

Samples of panettone sourdoughs manufactured in an industrial bakery located in Japan were investigated. The sourdough starter called mother sponge was imported from Italy and propagated continuously in Japan. According to traditional practice in Italy, panettone sourdoughs were prepared. Microflora was composed of Yeast (106-107cfu/g) and LAB (107-108cfu/g). All the fermentation process, predominant yeasts has the feature of no maltose fermentation and no growth at 37°C. Predominant LAB fermented maltose and has requirement of Tween 80 for growth. DNA-DNA hybridization data exhibited they were Candida milleri and Lactobacillus sanfranciscensis. Furthermore L. sanfranciscensis forming L-lactate was isolated from panettone sourdough. And lactate dehydrogenases took part in isomer of lactate produced by them. It was confirmed that the native microflora of the panettone mother sponge was successfully preserved in Japan.

Reclassification of Pediococcus urinaeequi (ex Mees 1934) Garvie 1988 as Aerococcus urinaeequi comb. nov.

The taxonomic status of Pediococcus urinaeequi is described, and the transfer of the species to the genus Aerococcus with the name Aerococcus urinaeequi comb. nov. is proposed, on the basis of the analysis of 16S rRNA gene sequence and DNA-DNA hybridization data.

Initial Subgingival Colonization of ‘Pristine’ Pockets

The treatment of periodontitis/peri-implantitis involves the reduction/eradication of periopathogens. After therapy, beneficial and pathogenic species recolonize the subgingival area. The dynamics of recolonization and especially the role of the supragingival environment in this process are still not well-understood. This prospective, split-mouth study followed the early colonization of 'pristine' pockets created during implant surgery (16 partially edentulous patients), to record the time needed before a complex subgingival flora could be established with the supragingival area as the single source. Four subgingival plaque samples were taken from shallow and medium pockets around implants (test), and neighboring teeth (undisturbed microbiota as reference) 1, 2, and 4 wks after abutment connection. Checkerboard DNA-DNA hybridization and culture data revealed a complex microbiota (including several pathogenic species) in the pristine pockets within a wk, with a minimal increase in counts up to 4 wks. Analysis of these data demonstrated that, even with the supragingival environment as the single source for colonizing bacteria, a complex subgingival microbiota can develop within 1 wk.

Emended description of porcine [Pasteurella] aerogenes, [Pasteurella] mairii and [Actinobacillus] rossii

The aim of this study was to improve the definition and identification of a group of veterinarily important bacteria referred to as the [Pasteurella] aerogenes-[Pasteurella] mairii-[Actinobacillus] rossii complex. These organisms have mainly been isolated from the reproductive and intestinal tracts of pigs and in most cases have been considered as opportunistic pathogens. A collection of 87 strains were characterized by phenotypic analysis from which 41 strains were selected for 16S rRNA gene sequence comparison, out of which 23 have been sequenced in the present study. One group of 21 strains phenotyped as biovars 1, 3-5, 9-11, 19 and 25-27, including the type strain of [P.] aerogenes, showed 16S rRNA gene sequence similarities of 99.6 % or higher; another group of 18 strains including biovars 2, 6-8, 12-15, 21, 23, 24 and 26A and the type strain of [A.] rossii showed 97.8 % or higher 16S rRNA gene sequence similarity. Between the two groups, 93.8-95.7 % 16S rRNA gene sequence similarity was observed. Strains of [P.] mairii showed 99.5 % similarity, with 95.5-97.2 and 93.8-95.5 % similarity to strains of [P.] aerogenes and [A.] rossii, respectively. Four strains could not be classified with any of these groups and belonged to other members of Pasteurellaceae. Comparisons were also made to DNA-DNA hybridization data. Biovars 1, 9, 10, 11 and 19, including the type strain of [P.] aerogenes, linked at 70 % DNA reassociation, whereas strains identified as biovars 2, 6, 7, 8, 12, 15 and 21 of [P.] aerogenes linked at 81 %. The latter group most likely represents [A.] rossii based on the 16S rRNA gene sequence comparisons. DNA reassociation between the [P.] aerogenes and [A.] rossii groups was at most 37 %, whereas 47 % was the highest DNA reassociation found between [P.] aerogenes and [P.] mairii. The study showed that [P.] aerogenes, [P.] mairii and [A.] rossii can not be easily separated and may consequently be misidentified based on current knowledge of their phenotypic characteristics. In addition, these taxa are difficult to separate from other taxa of the Pasteurellaceae. A revised scheme for separation based upon phenotypic characters is suggested for the three species [P.] aerogenes emend., [P.] mairii emend. and [A.] rossii emend., with the respective type strains ATCC 27883T, NCTC 10699T and ATCC 27072T.

Nicoletella semolina gen. nov., sp. nov., a New Member of Pasteurellaceae Isolated from Horses with Airway Disease

Gram-negative, nonmotile bacteria that are catalase, oxidase, and urease positive are regularly isolated from the airways of horses with clinical signs of respiratory disease. On the basis of the findings by a polyphasic approach, we propose that these strains be classified as Nicoletella semolina gen. nov, sp. nov., a new member of the family Pasteurellaceae. N. semolina reduces nitrate to nitrite but is otherwise biochemically inert; this includes the lack of an ability to ferment glucose and other sugars. Growth is fastidious, and the isolates have a distinctive colony morphology, with the colonies being dry and waxy and looking like a semolina particle that can be moved around on an agar plate without losing their shape. DNA-DNA hybridization data and multilocus phylogenetic analysis, including 16S rRNA gene (rDNA), rpoB, and infB sequencing, clearly placed N. semolina as a new genus in the family Pasteurellaceae. In all the phylogenetic trees constructed, N. semolina is on a distinct branch displaying approximately 5% 16S rDNA, approximately 16% rpoB, and approximately 20% infB sequence divergence from its nearest relative within the family Pasteurellaceae. High degrees of conservation of the 16S rDNA (99.8%), rpoB (99.6%), and infB (99.7%) sequences exist within the species, indicating that N. semolina isolates not only are phenotypically homogeneous but also are genetically homogeneous. The type strain of N. semolina is CCUG43639(T) (DSM16380(T)).