INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY logo - Volume 64, Issue Pt_2, 2014

Detection, identification and classification of yeasts have undergone a major transformation in the past decade and a half following application of gene sequence analyses and genome comparisons. Development of a database (barcode) of easily determined gene sequences from domains 1 and 2 (D1/D2) of large subunit rRNA and from the internal transcribed spacer (ITS) now permits many laboratories to identify species accurately and this has led to a doubling in the number of known species of yeasts over the past decade. Phylogenetic analysis of gene sequences has resulted in major revision of yeast systematics, resulting in redefinition of nearly all genera. Future work calls for application of genomics to refine our understanding of the species concept and to provide a better understanding of the boundaries of genera and higher levels of classification. This increased understanding of phylogeny is expected to allow prediction of the genetic potential of various clades and species for biotechnological applications and adaptation to environmental changes.

First-generation Sanger DNA sequencing revolutionized science over the past three decades and the current next-generation sequencing (NGS) technology has opened the doors to the next phase in the sequencing revolution. Using NGS, scientists are able to sequence entire genomes and to generate extensive transcriptome data from diverse photosynthetic eukaryotes in a timely and cost-effective manner. Genome data in particular shed light on the complicated evolutionary history of algae that form the basis of the food chain in many environments. In the Eukaryotic Tree of Life, the fact that photosynthetic lineages are positioned in four supergroups has important evolutionary consequences. We now know that the story of eukaryotic photosynthesis unfolds with a primary endosymbiosis between an ancestral heterotrophic protist and a captured cyanobacterium that gave rise to the glaucophytes, red algae and Viridiplantae (green algae and land plants). These primary plastids were then transferred to other eukaryotic groups through secondary endosymbiosis. A red alga was captured by the ancestor(s) of the stramenopiles, alveolates (dinoflagellates, apicomplexa, chromeridae), cryptophytes and haptophytes, whereas green algae were captured independently by the common ancestors of the euglenophytes and chlorarachniophytes. A separate case of primary endosymbiosis is found in the filose amoeba Paulinella chromatophora, which has at least nine heterotrophic sister species. Paulinella genome data provide detailed insights into the early stages of plastid establishment. Therefore, genome data produced by NGS have provided many novel insights into the taxonomy, phylogeny and evolutionary history of photosynthetic eukaryotes.

Among available genome relatedness indices, average nucleotide identity (ANI) is one of the most robust measurements of genomic relatedness between strains, and has great potential in the taxonomy of bacteria and archaea as a substitute for the labour-intensive DNA–DNA hybridization (DDH) technique. An ANI threshold range (95–96 %) for species demarcation had previously been suggested based on comparative investigation between DDH and ANI values, albeit with rather limited datasets. Furthermore, its generality was not tested on all lineages of prokaryotes. Here, we investigated the overall distribution of ANI values generated by pairwise comparison of 6787 genomes of prokaryotes belonging to 22 phyla to see whether the suggested range can be applied to all species. There was an apparent distinction in the overall ANI distribution between intra- and interspecies relationships at around 95–96 % ANI. We went on to determine which level of 16S rRNA gene sequence similarity corresponds to the currently accepted ANI threshold for species demarcation using over one million comparisons. A twofold cross-validation statistical test revealed that 98.65 % 16S rRNA gene sequence similarity can be used as the threshold for differentiating two species, which is consistent with previous suggestions (98.2–99.0 %) derived from comparative studies between DDH and 16S rRNA gene sequence similarity. Our findings should be useful in accelerating the use of genomic sequence data in the taxonomy of bacteria and archaea.

The G+C content of a genome is frequently used in taxonomic descriptions of species and genera. In the past it has been determined using conventional, indirect methods, but it is nowadays reasonable to calculate the DNA G+C content directly from the increasingly available and affordable genome sequences. The expected increase in accuracy, however, might alter the way in which the G+C content is used for drawing taxonomic conclusions. We here re-estimate the literature assumption that the G+C content can vary up to 3–5 % within species using genomic datasets. The resulting G+C content differences are compared with DNA–DNA hybridization (DDH) similarities calculated in silico using the GGDC web server, with 70 % similarity as the gold standard threshold for species boundaries. The results indicate that the G+C content, if computed from genome sequences, varies no more than 1 % within species. Statistical models based on larger differences alone can reject the hypothesis that two strains belong to the same species. Because DDH similarities between two non-type strains occur in the genomic datasets, we also examine to what extent and under which conditions such a similarity could be <70 % even though the similarity of either strain to a type strain was ≥70 %. In theory, their similarity could be as low as 50 %, whereas empirical data suggest a boundary closer (but not identical) to 70 %. However, it is shown that using a 50 % boundary would not affect the conclusions regarding the DNA G+C content. Hence, we suggest that discrepancies between G+C content data provided in species descriptions on the one hand and those recalculated after genome sequencing on the other hand ≥1 % are due to significant inaccuracies of the applied conventional methods and accordingly call for emendations of species descriptions.

Vibrios are ubiquitous in the aquatic environment and can be found in association with animal or plant hosts. The range of ecological relationships includes pathogenic and mutualistic associations. To gain a better understanding of the ecology of these microbes, it is important to determine their phenotypic features. However, the traditional phenotypic characterization of vibrios has been expensive, time-consuming and restricted in scope to a limited number of features. In addition, most of the commercial systems applied for phenotypic characterization cannot characterize the broad spectrum of environmental strains. A reliable and possible alternative is to obtain phenotypic information directly from whole genome sequences. The aim of the present study was to evaluate the usefulness of whole genome sequences as a source of phenotypic information. We performed a comparison of the vibrio phenotypes obtained from the literature with the phenotypes obtained from whole genome sequences. We observed a significant correlation between the previously published phenotypic data and the phenotypic data retrieved from whole genome sequences of vibrios. Analysis of 26 vibrio genomes revealed that all genes coding for the specific proteins involved in the metabolic pathways responsible for positive phenotypes of the 14 diagnostic features (Voges–Proskauer reaction, indole production, arginine dihydrolase, ornithine decarboxylase, utilization of myo-inositol, sucrose and l-leucine, and fermentation of d-mannitol, d-sorbitol, l-arabinose, trehalose, cellobiose, d-mannose and d-galactose) were found in the majority of the vibrios genomes. Vibrio species that were negative for a given phenotype revealed the absence of all or several genes involved in the respective biochemical pathways, indicating the utility of this approach to characterize the phenotypes of vibrios. The absence of the global regulation and regulatory proteins in the Vibrio parahaemolyticus genome indicated a non-vibrio phenotype. Whole genome sequences represent an important source for the phenotypic identification of vibrios.

Genome sequences are enabling applications of different approaches to more clearly understand microbial phylogeny and systematics. Two of these approaches involve identification of conserved signature indels (CSIs) and conserved signature proteins (CSPs) that are specific for different lineages. These molecular markers provide novel and more definitive means for demarcation of prokaryotic taxa and for identification of species from these groups. Genome sequences are also enabling determination of phylogenetic relationships among species based upon sequences for multiple proteins. In this work, we have used all of these approaches for studying the phytopathogenic bacteria belonging to the genera Dickeya , Pectobacterium and Brenneria . Members of these genera, which cause numerous diseases in important food crops and ornamental plants, are presently distinguished mainly on the basis of their branching in phylogenetic trees. No biochemical or molecular characteristic is known that is uniquely shared by species from these genera. Hence, detailed studies using the above approaches were carried out on proteins from the genomes of these bacteria to identify molecular markers that are specific for them. In phylogenetic trees based upon concatenated sequences for 23 conserved proteins, members of the genera Dickeya , Pectobacterium and Brenneria formed a strongly supported clade within the other Enterobacteriales . Comparative analysis of protein sequences from the Dickeya , Pectobacterium and Brenneria genomes has identified 10 CSIs and five CSPs that are either uniquely or largely found in all genome-sequenced species from these genera, but not present in any other bacteria in the database. In addition, our analyses have identified 10 CSIs and 17 CSPs that are specifically present in either all or most sequenced Dickeya species/strains, and six CSIs and 19 CSPs that are uniquely found in the sequenced Pectobacterium genomes. Finally, our analysis also identified three CSIs and one CSP that are specifically shared by members of the genera Pectobacterium and Brenneria , but absent in species of the genus Dickeya , indicating that the former two genera shared a common ancestor exclusive of Dickeya . The identified CSIs and CSPs provide novel tools for identification of members of the genera Dickeya and Pectobacterium and for delimiting these taxa in molecular terms. Descriptions of the genera Dickeya and Pectobacterium have been revised to provide information for these molecular markers. Biochemical studies on these CSIs and CSPs, which are specific for these genera, may lead to discovery of novel properties that are unique to these bacteria and which could be targeted to develop antibacterial agents that are specific for these plant-pathogenic bacteria.

Currently, bacterial taxonomy relies on a polyphasic approach based on the combination of phenotypic and genotypic characteristics. However, the current situation is paradoxical in that the genetic criteria that are used, including DNA–DNA hybridization, 16S rRNA gene sequence nucleotide similarity and phylogeny, and DNA G+C content, have significant limitations, but genome sequences that contain the whole genetic information of bacterial strains are not used for taxonomic purposes, despite the decreasing costs of sequencing and the increasing number of available genomes. Recently, we diversified bacterial culture conditions with the aim of isolating uncultivated bacteria. To classify the putative novel species that we cultivated, we used a polyphasic strategy that included phenotypic as well as genomic criteria (genome characteristics as well as genomic sequence similarity). Herein, we review the pros and cons of genome sequencing for taxonomy and propose that the incorporation of genome sequences in taxonomic studies has the advantage of using reliable and reproducible data. This strategy, which we name taxono-genomics, may contribute to the taxonomic classification of bacteria.

A psychrotolerant methanogenic strain, X-18T, was isolated from the soil of the Madoi wetland at Qinghai, Tibetan plateau, China. Cells were wavy rods (11–62 μm long) with blunt tapered ends and Gram-stain-negative. Strain X-18T grew strictly anaerobically and produced methane exclusively from H2/CO2. Growth occurred in the temperature range of 4–32 °C and optimally at 25 °C. Growth pH ranged from 6.5 to 8.0 and the optimum was 7.0. The G+C content of the genomic DNA of strain X-18T was 44.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequences and the alpha subunit of methyl-coenzyme M reductase indicated that strain X-18T was affiliated to the genus Methanospirillum and was most closely related to Methanospirillum lacunae Ki8-1T, with 96.3 % 16S rRNA gene sequence similarity. However, strain X-18T could be distinguished from the existing species of the genus Methanospirillum by its lower growth temperature and obligate hydrogenotrophic methanogenesis. On the basis of phenotypic characteristics and phylogenetic analysis, strain X-18T represents a novel species of the genus Methanospirillum , for which the name Methanospirillum psychrodurum sp. nov. is proposed and strain X-18T is assigned as the type strain ( = CGMCC 1.5186T = JCM 19216T).

A Gram-stain-positive, flagellate, rod-shaped, catalase- and oxidase-positive bacterium, designated DCY72T, was isolated from the soil of a Gynostemma pentaphyllum field. Growth occurred at 4–34 °C (optimum 30 °C), at pH 4–10 (optimum pH 7), and with 0–5 % NaCl (w/v). The major menaquinones of strain DCY72T were MK-9(H2) (81.0 %) and MK-10(H2) (12.2 %). The major amino acid present in the cell-wall peptidoglycan was l-lysine. The major fatty acids were anteiso-C15 : 0 and anteiso-C17 : 0. The genomic DNA G+C content was 64.5 mol%. 16S rRNA gene sequence analysis revealed that strain DCY72T belonged to the family Micrococcaceae and was most closely related to Arthrobacter ramosus CCM 1646T (98.2 % similarity). The DNA–DNA relatedness between strain DCY72T and A. ramosus KACC 14391T (98.2 % 16S rRNA gene sequence similarity), Arthrobacter nitroguajacolicus KACC 14581T (97.6 %), Arthrobacter nicotinovorans KACC 20508T (97.3 %) and Arthrobacter aurescens KACC 20528T (97.3 %). was 12.9 %±0.3, 25.6 %±0.3, 26.6 %±0.5 and 23.2 %±0.9, respectively. On the basis of the phenotypic characteristics, genotypic analysis and physiological characteristics, strain DCY72T represents a novel species of the genus Arthrobacter , for which the name Arthrobacter gyeryo ngensis sp. nov. is proposed. The type strain is DCY72T ( = KCTC 33072T = JCM 18514T).

Gram-staining-positive anaerobic rods were isolated from the faeces of a wild lowland gorilla (Gorilla gorilla gorilla) in Moukalaba-Doudou National Park, Gabon, and strain GG01T was taxonomically investigated. Based on phylogenetic analyses and specific phenotypic characteristics, the strain belonged to the genus Bifidobacterium . Phylogenetic analysis of its 16S rRNA gene sequence revealed that strain GG01T formed a single monophyletic cluster and had a distinct line of descent. Based on 16S rRNA gene sequence similarity, the type strains of Bifidobacterium catenulatum JCM 1194T (98.3 %) and Bifidobacterium pseudocatenulatum (98.1 %) JCM 1200T were the most closely related to this novel strain, although it was clear that they belonged to different species. hsp60 sequences also supported these relationships. The DNA G+C content of this novel strain was 60.1 mol%. Bifidobacterium moukalabense sp. nov. (type strain GG01T = JCM 18751T = DSM 27321T) is proposed.

A novel, Gram-staining-positive, facultatively anaerobic, non-motile and coccus-shaped bacterium, strain WL80T, was isolated from the gut of an abalone, Haliotis discus hannai, collected from the northern coast of Jeju in Korea. Optimal growth occurred at 30 °C, pH 7–8 and with 1 % (w/v) NaCl. Phylogenetic analyses based on the 16S rRNA gene sequence revealed that strain WL80T fell within the cluster of the genus Actinomyces , with highest sequence similarity to the type strains of Actinomyces radicidentis (98.8 % similarity) and Actinomyces urogenitalis (97.0 % similarity). The major cellular fatty acids were C18 : 1ω9c and C16 : 0. Menaquinone-10 (H4) was the major respiratory quinone. The genomic DNA G+C content of the isolate was 70.4 mol%. DNA–DNA hybridization values with closely related strains indicated less than 7.6 % genomic relatedness. The results of physiological, biochemical, chemotaxonomic and genotypic analyses indicated that strain WL80T represents a novel species of the genus Actinomyces , for which the name Actinomyces haliotis sp. nov. is proposed. The type strain is WL80T ( = KACC 17211T = JCM 18848T).

A novel actinomycete, designated strain NEAU-ycm2T, was isolated from edible Chinese black ants (Polyrhachis vicina Roger) and characterized using a polyphasic approach. The organism was found to have morphological and chemotaxonomic characteristics typical of the genus Micromonospora . The 16S rRNA gene sequence of strain NEAU-ycm2T showed highest similarity to those of Micromonospora sonneratiae 274745T (99.12 %), Micromonospora pattaloongensis TJ2-2T (98.85 %), Micromonospora pisi GUI 15T (98.76 %), Polymorphospora rubra TT 97-42T (98.42 %) and Micromonospora eburnea LK2-10T (98.21 %). Phylogenetic analysis based on the 16S rRNA gene and gyrB gene demonstrated that strain NEAU-ycm2T is a member of the genus Micromonospora and supported the close phylogenetic relationship to M. sonneratiae 274745T, M. pattaloongensis JCM 12833T and M. pisi GUI 15T. Furthermore, a combination of DNA–DNA hybridization and some physiological and biochemical properties indicated that the novel strain could be readily distinguished from its closest phylogenetic relatives. Therefore, it is proposed that NEAU-ycm2T represents a novel species of the genus of Micromonospora , for which the name Micromonospora polyrhachis sp. nov. is proposed. The type strain is NEAU-ycm2T ( = CGMCC 4.7100T = DSM 45886T).

A Gram-stain-positive, novel actinobacterium, designated strain JL-6T, was isolated from the litter of a bamboo (Sasa borealis) forest in Damyang, Korea. Strain JL-6T had white‐grey, smooth, cylindrical spores that were borne in straight, long spore-chains. The novel strain grew aerobically at 15–28 °C (optimum, 28 °C), pH 4.0–8.0 (optimum, pH 5.5) and with 0–1.5 % (w/v) NaCl. The cell-wall peptidoglycan contained ll-diaminopimelic acid, glutamic acid, alanine and glycine. The predominant menaquinones were MK-9(H6) and MK-9(H8). Whole-cell hydrolysates mainly contained glucose and ribose. Phosphatidylinositol and phosphatidylcholine were the diagnostic phospholipids. The G+C content of the genomic DNA was 72.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JL-6T belonged to the genus Streptomyces with sequence similarities ranging from 97.3 % to 98.3 %. However, DNA–DNA hybridization between JL-6T and the closest related strain, Streptomyces turgidiscabies , ATCC 700248T and other closely related species in the genus Streptomyces showed <50 % relatedness. Based on these observations, strain JL-6T is proposed to represent a novel species of the genus Streptomyces , for which the name Streptomyces graminilatus sp. nov. is proposed. The type strain is JL-6T ( = KACC 16470T = NBRC 108882T).

Bacterial strains KIS82-1T and GRS42T were isolated from soil and from sap of Acer mono, respectively, in the Republic of Korea. Both strains were aerobic, Gram-stain-positive, mesophilic, rod-shaped and motile. Phylogenetically, both strains belonged to the family Microbacteriaceae of the phylum Actinobacteria . The 16S rRNA gene sequence of strain KIS82-1T showed the highest similarity to those of Frondihabitans peucedani RS-15T (97.6 %), Frigoribacterium mesophilum MSL-08T (97.2 %) and Labedella gwakjiensis KSW2-17T (97.0 %), while strain GRS42T showed the highest 16S rRNA gene sequence similarity to Frondihabitans peucedani RS-15T (98.7 %), Frondihabitans cladoniiphilus CafT13T (98.4 %), Frondihabitans australicus E1HC-02T (98.2 %) and Frigoribacterium faeni 801T (97.3 %). The 16S rRNA gene sequence similarity between GRS42T and KIS82-1T was 97.0 %. Phylogenetic trees indicated that strain GRS42T was firmly grouped into the genus Frondihabitans , while strain KIS82-1T did not show a clear affiliation to any genus within the family Microbacteriaceae . Strain KIS82-1T showed type B1β peptidoglycan with 2,4-diamino-l-butyric acid as the diamino acid. It had MK-11, MK-10 and MK-12 as respiratory quinones, anteiso-C15 : 0, iso-C16 : 0 and iso-C14 : 0 as major cellular fatty acids and diphosphatidylglycerol, phosphatidylglycerol and an unknown glycolipid as predominant polar lipids. The peptidoglycan of strain GRS42T was of type B2β with d-ornithine as the diamino acid. The strain contained MK-8, MK-9 and MK-7 as respiratory quinones, summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c) as major cellular fatty acid and diphosphatidylglycerol, phosphatidylglycerol and three unknown glycolipids as predominant polar lipids. Strain GRS42T revealed low DNA–DNA hybridization (<50 % relatedness) with closely related strains. Based on the data obtained in the present polyphasic taxonomic study, we propose that strain KIS82-1T represents a novel genus and species and that strain GRS42T represents a novel species in the family Microbacteriaceae . The genus Galbitalea gen. nov. is proposed, with strain KIS82-1T ( = KACC 15520T = NBRC 108727T) as the type strain of the type species, Galbitalea soli sp. nov. Strain GRS42T ( = KACC 15521T = NBRC 108728T) is proposed as the type strain of Frondihabitans sucicola sp. nov.

A Gram-stain-positive, aerobic, non-spore-forming, short-rod-shaped bacterium, designated strain MP203T, was isolated from ice water of Midui Glacier in Tibet Autonomous Region, China. The strain was psychrotolerant, growing at 0–25 °C. 16S rRNA gene sequence analysis showed that strain MP203T was most similar to Frigoribacterium faeni NBRC 103066T, Compostimonas suwonensis KACC 13354T, Frigoribacterium mesophilum KCTC 19311T, Marisediminicola antarctica CCTCC AB 209077T and Alpinimonas psychrophila JCM 18951T, with similarities of 97.4, 97.2, 97.2, 97.1 and 97.1 %, respectively. The maximum-likelihood phylogenetic tree indicated that strain MP203T clustered with nine genera of the family Microbacteriaceae , namely Frigoribacterium , Compostimonas , Marisediminicola , Alpinimonas , Frondihabitans , Clavibacter , Subtercola , Klugiella and Agreia . However, bootstrap analysis showed that there was no significance in the branching pattern of the linage comprising strain MP203T and any existing generic lineage of the family Microbacteriaceae . DNA–DNA hybridization results indicated levels of relatedness between strain MP203T and Marisediminicola antarctica CCTCC AB 209077T, Frigoribacterium faeni NBRC 103066T, Frigoribacterium mesophilum KCTC 19311T, Compostimonas suwonensis KACC 13354T and Alpinimonas psychrophila JCM 18951T were 25.8±7.3, 29.6±7.6, 19.7±6.7, 16.0±4.2 and 12.4±5.1 % (mean±sd), respectively. The G+C content of the genomic DNA was 64.1 mol%. Analysis of the cell-wall peptidoglycan revealed that the peptidoglycan structure of strain MP203T was B10 type with Gly[l-Hse]–d-Glu–d-DAB, containing 2, 4-diaminobutyric acid (DAB) as a diagnostic amino acid. The cell-wall sugars were rhamnose, ribose, mannose and glucose. The major fatty acids were anteiso-C15 : 0, iso-C16 : 0 and anteiso A-C15 : 1. An unusual compound identified as anteiso-C15 : 0-DMA (1, 1-dimethoxy-anteiso-pentadecane) was also present in strain MP203T. The predominant menaquinone was MK-10. Diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), one unknown glycolipid and four unknown lipids were detected in the polar lipid extracts. As strain MP203T was distinguishable from phylogenetically related genera in the family Microbacteriaceae in terms of its physiological and chemotaxonomic characteristics and phylogenetic position, it was considered to represent a novel species of a new genus. Thus, the name Glaciihabitans tibetensis gen. nov., sp. nov. is proposed. The type strain of Glaciihabitans tibetensis is MP203T ( = CGMCC 1.12484T = KCTC 29148T).

In 2000, an actinomycete strain that showed strong antibacterial activity in culture extracts was isolated from a soil sample. The antibiotic activity corresponds to a lipopeptide complex that was named friulimycin, as the producing micro-organism was isolated from a soil sample from the region of Friaul in Italy. Taxonomic investigations showed that the producer strain belonged to a novel species of the genus Actinoplanes , for which the name Actinoplanes friuliensis was proposed. During further taxonomic studies, another antibiotic-producing isolate belonging to the genus Actinoplanes , FH 2241T, was characterized; in a patent, the name ‘Actinoplanes nipponensis’ was proposed for this strain. This organism was shown to be related to A. friuliensis . ‘A. nipponensis’ was never described in detail and the name was never validly published. Here we present a complete description of Actinoplanes nipponensis sp. Nov. (type strain FH 2241T = ATCC 31145T = DSM 43867T) and an emended description of Actinoplanes friuliensis (type strain HAG 010964T = DSM 45797T = CCUG 63250T).

Two strains, designated K2814T and K282, were isolated from a compost pile in Japan. These strains were Gram-stain-variable, aerobic, motile and endospore-forming rods. The strains produced a characteristic brown non-diffusible pigment. The 16S rRNA gene sequences of the strains were 100 % identical and had high similarity to that of Brevibacillus levickii LMG 22481T (97.3 %). Phylogenetic analyses based on 16S rRNA gene sequences revealed that these strains belong to the genus Brevibacillus . Strains K2814T and K282 contained meso-diaminopimelic acid in their cell walls. Strains K2814T and K282 contained MK-7 (96.0 and 97.2 %, respectively) and MK-8 (4.0 and 2.8 %, respectively) as the major and minor menaquinones, respectively. Their major cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0 and iso-C17 : 0. The DNA G+C contents of strains K2814T and K282 were 48.8 and 49.8 mol%, respectively. Polar lipids of strain K2814T were composed of phosphatidyl-N-methylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, an unidentified phospholipid, three unidentified polar lipids, an unidentified aminophospholipid and an unidentified aminolipid. The level of DNA–DNA relatedness between strains K2814T and K282 was 99 or 100 %, and levels between strain K2814T and the type strains of seven related species of the genus Brevibacillus , including Brevibacillus levickii LMG 22481T, were below 59 %. From the chemotaxonomic and physiological data and the levels of DNA–DNA relatedness, these two strains should be classified as representing a novel species of the genus Brevibacillus , for which the name Brevibacillus fulvus sp. nov. (type strain K2814T = JCM 18162T = ATCC BAA-2417T = DSM 25523T) is proposed.