Comprehensive lipidomic analysis of the genus Cutibacterium.

Cutibacterium are part of the human skin microbiota and are opportunistic microorganisms that become pathogenic in immunodeficient states. These lipophilic bacteria willingly inhabit areas of the skin where sebaceous glands are abundant; hence, there is a need to thoroughly understand their metabolism. Lipids are no longer considered only structural elements but also serve as signaling molecules and may have antigenic properties. Lipidomics remains a major research challenge, mainly due to the diverse physicochemical properties of lipids. Therefore, this study aimed to perform a large comparative lipidomic analysis of eight representatives of the Cutibacterium genus, including four phylotypes of C. acnes and two strains of C. granulosum, C. avidum, and C. namnetense. Lipidomic analysis was performed by liquid chromatography‒mass spectrometry (LC-MS) in both positive and negative ion modes, allowing the detection of the widest range of metabolites. Fatty acid analysis by gas chromatography‒mass spectrometry (GC-MS) corroborated the lipidomic data. As a result, 128 lipids were identified, among which it was possible to select marker compounds, some of which were characteristic even of individual C. acnes phylotypes. These include phosphatidylcholine PC 30:0, sphingomyelins (SM 33:1, SM 35:1), and phosphatidylglycerol with an alkyl ether substituent PG O-32:0. Moreover, cardiolipins and fatty acid amides were identified in Cutibacterium spp. for the first time. This comparative characterization of the cutibacterial lipidome with the search for specific molecular markers reveals its diagnostic potential for clinical microbiology. IMPORTANCE: Cutibacterium (previously Propionibacterium) represents an important part of the human skin microbiota, and its role in clinical microbiology is growing due to opportunistic infections. Lipidomics, apart from protein profiling, has the potential to prove to be a useful tool for defining the cellular fingerprint, allowing for precise differentiation of microorganisms. In this work, we presented a comparative analysis of lipids found in eight strains of the genus Cutibacterium, including a few C. acnes phylotypes. Our results are one of the first large-scale comprehensive studies regarding the bacterial lipidome, which also enabled the selection of C. acnes phylotype-specific lipid markers. The increased role of lipids not only as structural components but also as diagnostic markers or potential antigens has led to new lipid markers that can be used as diagnostic tools for clinical microbiology. We believe that the findings in our paper will appeal to a wide range of researchers.

Acceptor-Reactivity-Controlled Stereoconvergent Synthesis and Immunological Activity of a Unique Pentasaccharide from the Cell Wall Polysaccharide of Cutibacterium acnes C7.

Bacterial cell-surface polysaccharides are involved in various biological processes and have attracted widespread attention as potential targets for developing carbohydrate-based drugs. However, the accessibility to structurally well-defined polysaccharide or related active oligosaccharide domains remains challenging. Herein, we describe an efficiently stereocontrolled approach for the first total synthesis of a unique pentasaccharide repeating unit containing four difficult-to-construct 1,2-cis-glycosidic linkages from the cell wall polysaccharide of Cutibacterium acnes C7. The features of our approach include: 1) acceptor-reactivity-controlled glycosylation to stereoselectively construct two challenging rare 1,2-cis-ManA2,3(NAc)2 (ß-2,3-diacetamido-2,3-dideoxymannuronic acid) linkages, 2) combination use of 6-O-tert-butyldiphenylsilyl (6-O-TBDPS)-mediated steric shielding effect and ether solvent effect to stereoselectively install a 1,2-cis-glucosidic linkage, 3) bulky 4,6-di-O-tert-butylsilylene (DTBS)-directed glycosylation to stereospecifically construct a 1,2-cis-galactosidic linkage, 4) stereoconvergent [2+2+1] and one-pot chemoselective glycosylation to rapidly assemble the target pentasaccharide. Immunological activity tests suggest that the pentasaccharide can induce the production of proinflammatory cytokine TNF-α in a dose-dependent manner.

Characterization of a Cutibacterium acnes Camp Factor 1-Related Peptide as a New TLR-2 Modulator in In Vitro and Ex Vivo Models of Inflammation.

Cutibacterium acnes (C. acnes) has been implicated in inflammatory acne where highly mutated Christie-Atkins-Munch-Petersen factor (CAMP)1 displays strong toll like receptor (TLR)-2 binding activity. Using specific antibodies, we showed that CAMP1 production was independent of C. acnes phylotype and involved in the induction of inflammation. We confirmed that TLR-2 bound both mutated and non-mutated recombinant CAMP1, and peptide array analysis showed that seven peptides (A14, A15, B1, B2, B3, C1 and C3) were involved in TLR-2 binding, located on the same side of the three-dimensional structure of CAMP1. Both mutated and non-mutated recombinant CAMP1 proteins induced the production of C-X-C motif chemokine ligand interleukin (CXCL)8/(IL)-8 in vitro in keratinocytes and that of granulocyte macrophage-colony stimulating factor (GM-CSF), tumor necrosis factor (TNF)-α, IL-1ß and IL-10 in ex vivo human skin explants. Only A14, B1 and B2 inhibited the production of CXCL8/IL-8 by keratinocytes and that of (GM-CSF), TNF-α, IL-1ß and IL-10 in human skin explants stimulated with rCAMP1 and C. acnes. Following pretreatment with B2, RNA sequencing on skin explants identified the 10 genes displaying the strongest differential expression as IL6, TNF, CXCL1, CXCL2, CXCL3, CXCL8, IL-1ß, chemokine ligand (CCL)2, CCL4 and colony stimulating factor (CSF)2. We, thus, identified a new CAMP1-derived peptide as a TLR-2 modulator likely to be a good candidate for clinical evaluation.

Increased immunogenicity and protection of recombinant Sm14 antigens by heat-killed Cutibacterium acnes in BALB/c mice infected with Schistosoma mansoni.

After many years of the excessive use of praziquantel against Schistosoma mansoni (S. mansoni), it has already led to the development of drug resistance. While schistosomiasis is still affecting millions of people every year, vaccination may be one realistic alternative way to control the disease. Currently, S. mansoni 14-kDa fatty acid-binding protein (Sm14) has shown promising results as a vaccine antigen. Yet, the use of an adjuvant may be necessary to further increase the effectiveness of the vaccine. Herein, we investigated the potential of using heat-killed Cutibacterium acnes (C. acnes) as an adjuvant for recombinant Sm14 (rSm14). Immunization of mice with C. acnes-adjuvanted rSm14 showed increased humoral immune responses, compared with mice immunized with rSm14 alone. Additionally, C. acnes-adjuvanted rSm14 vaccination provided higher protection to mice against S. mansoni infection and liver injuries. These results suggest that C. acnes increases the immunogenicity of rSm14, which leads to better protection against S. mansoni infection. Therefore, heat-killed C. acnes may be a promising adjuvant to use with rSm14.

Antibiofilm activity of Cutibacterium acnes cell-free conditioned media against Staphylococcus spp.

Staphylococcus spp. and Cutibacterium acnes are members of the skin microbiome but can also act as pathogens. Particularly, Staphylococcus species are known to cause medical devices-associated infections, and biofilm production is one of their main virulence factors. Biofilms allow bacteria to adhere and persist on surfaces, protecting them from antimicrobials and host defenses. Since both bacteria are found in the human skin, potentially competing for niches, we aimed to investigate if C. acnes produces molecules that affect Staphylococcus spp. biofilm formation and dispersal. Thus, we evaluated the impact of C. acnes cell-free conditioned media (CFCM) on S. aureus, S. epidermidis, S. hominis, and S. lugdunensis biofilm formation. S. lugdunensis and S. hominis biofilm formation was significantly reduced with C. acnes CFCM without impact on their planktonic growth. C. acnes CFCM also significantly disrupted S. hominis established biofilms. The active molecules against S. lugdunensis and S. hominis biofilms appeared to be distinct since initial characterization points to different sizes and sensitivity to sodium metaperiodate, although the activity is highly resistant to heat in both cases. Mass spectrometry analysis of the fractions active against S. hominis revealed several potential candidates. Investigating how species present in the same environment interact, affecting the dynamics of biofilm formation, may reveal clinically useful compounds as well as molecular aspects of interspecies interactions.

Propionibacterium/Cutibacterium species-related positive samples, identification, clinical and resistance features: a 10-year survey in a French hospital.

A 10-year retrospective study of Propionibacterium/Cutibacterium-positive samples gathered from hospitalized patients was conducted at Nantes University hospital. A total of 2728 Propionibacterium/Cutibacterium-positive samples analyzed between 2007 and 2016 were included. Due to the implementation of MALDI-TOF identification in 2013, most non-Cutibacterium acnes isolates were identified a second time using this technology. Over that period, Cutibacterium acnes remained the most predominant species accounting for 91.5% (2497/2728) of the isolates, followed by Cutibacterium avidum (4.2%, 115/2728) and Cutibacterium granulosum (2.4%, 64/2728). Regarding the origin of samples, the orthopaedic department was the main Cutibacterium sample provider representing 51.9% (1415/2728) of all samples followed by the dermatology department (11.5%, 315/2728). Samples were recovered from various tissue locations: 31.5% (858/2728) from surgery-related samples such as shoulder, spine or hip replacement devices and 19.1% (520/2728) from skin samples. MALDI-TOF method revealed misidentification before 2013. Cutibacterium avidum was falsely identified as C. granulosum (n = 33). Consequently, MALDI-TOF technology using up-to-date databases should be preferred to biochemical identification in order to avoid biased species identification. Regarding antibiotic resistance, 14.7% (20/136) of C. acnes was resistant to erythromycin. 4.1% (41/1005) of C. acnes strains, 17.9% (12/67) of C. avidum strains and 3.6% (1/28) of C. granulosum strains were found resistant to clindamycin.

Microlunatus terrae sp. nov., a bacterium isolated from soil.

Strain BS6(T), a Gram-positive non-motile bacterium, was isolated from soil in South Korea and characterized to determine its taxonomic position. Phylogenetic analyses based on the 16S rRNA gene sequence revealed that strain BS6T belonged to the family Propionibacteriaceae in the class Actinobacteria. Strain BS6(T) showed the highest 16S rRNA gene sequence similarity with Microlunatus soli CC-012602(T) (98.6%) and high sequence similarities with Microlunatus species (94.5-98.6%). Chemotaxonomic data revealed that the predominant fatty acids were anteiso-C(17:0), anteiso-C(15:0), summed feature 8 (C(18:1) ω7c/ω6c), and iso-C(16:0). The cell wall peptidoglycan contained (LL)-diaminopimelic acid, and the major polar lipids were diphosphatidylglycerol, and phosphatidylglycerol. Based on these data, BS6(T) (=KCTC 19858(T) =JCM 17661(T) =CCARM 9244(T) =KEMC 9004-079(T)) should be classified as a type strain of a novel species, for which the name Microlunatus terrae sp. nov. is proposed.

Microlunatus parietis sp. nov., isolated from an indoor wall.

A Gram-positive, coccoid, non-endospore-forming actinobacterium (strain 12-Be-011(T)) was isolated from indoor wall material. Based on 16S rRNA gene sequence comparisons, strain 12-Be-011(T) was clearly shown to belong to the genus Microlunatus and was most closely related to Microlunatus panaciterrae Gsoil 954(T) (95.7 %), Microlunatus soli CC-12602(T) (94.9 %), Microlunatus ginsengisoli Gsoil 633(T) (94.8 %), Microlunatus aurantiacus YIM 45721(T) (95.5 %) and Microlunatus phosphovorus DSM 10555(T) (94.7 %). The cell-wall peptidoglycan contained LL-diaminopimelic acid as the diagnostic diamino acid. Mycolic acids were absent. The major menaquinone was MK-9(H4). The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, two unknown phospholipids and one unknown glycolipid. The major fatty acids of iso-C15:0, anteiso-C15:0 and iso-C16:0 supported the affiliation of strain 12-Be-011(T) to the genus Microlunatus. Physiological and biochemical test results allowed a clear phenotypic differentiation of strain 12-Be-011(T) from all other species of the genus Microlunatus. Hence, strain 12-Be-011(T) can be regarded as a representative of a novel species, for which the name Microlunatus parietis sp. nov. is proposed, with the type strain 12-Be-011(T) (=DSM 22083(T)=CCM 7636(T)).

Microlunatus soli sp. nov., isolated from soil.

A Gram-stain-positive, coccoid, non-endospore-forming actinobacterium (strain CC-12602(T)) was isolated from a spawn used for growing the edible mushroom Agaricus brasiliensis in the laboratory. On the basis of 16S rRNA gene sequence analysis, strain CC-12602(T) was shown to belong to the genus Microlunatus and was related most closely to the type strains of Microlunatus ginsengisoli (96.1 % similarity), M. phosphovorus (95.9 %), M. panaciterrae (95.8 %) and M. aurantiacus (95.5 %). The quinone system comprised menaquinone MK-9(H4) as the major component and the polyamine pattern consisted of spermidine and spermine as major compounds. The predominant polar lipids were phosphatidylglycerol and unknown phospholipid PL3. Moderate amounts of diphosphatidylglycerol, an unknown glycolipid and three unknown phospholipids and minor amounts of an unknown phospholipid and a polar lipid were detected. The peptidoglycan type was A3gamma', based on LL-2,6-diaminopimelic acid with an interpeptide bridge consisting of a single glycine residue and a second glycine residue at position 1 of the peptide subunit. Peptidoglycan structure and major fatty acids (anteiso-C(15 : 0), iso-C(16 : 0) and iso-C(15 : 0)) supported the affiliation of strain CC-12602(T) to the genus Microlunatus. The results of physiological and biochemical tests allowed strain CC-12602(T) to be differentiated phenotypically from recognized Microlunatus species. Strain CC-12602(T) is therefore considered to represent a novel species of the genus Microlunatus, for which the name Microlunatus soli sp. nov. is proposed. The type strain is CC-12602(T) (=DSM 21800(T) =CCM 7685( T)).

Microlunatus panaciterrae sp. nov., a beta-glucosidase-producing bacterium isolated from soil in a ginseng field.

A novel bacterial strain, designated Gsoil 954(T), showing beta-glucosidase activity was isolated from a soil sample from a ginseng field in Pocheon Province, South Korea. The isolate was a Gram-positive, aerobic, motile, coccus-shaped, non-endospore-forming bacterium. Comparative analysis of 16S rRNA gene sequences revealed that strain Gsoil 954(T) belonged to the family Propionibacteriaceae, the highest levels of sequence similarity being found with the type strains of Microlunatus ginsengisoli (96.4 %) and Microlunatus phosphovorus (96.2 %). The strain showed <95.9 % similarity with respect to other species within the family Propionibacteriaceae. In addition, the presence of ll-2,6-diaminopimelic acid in the cell-wall peptidoglycan, MK-9(H(4)) as the major menaquinone and anteiso-C(15 : 0) and iso-C(15 : 0) as the major cellular fatty acids suggested its affiliation with the genus Microlunatus. The G+C content of the genomic DNA was 65.1 mol%. On the basis of its phenotypic characteristics and phylogenetic distinctiveness, strain Gsoil 954(T) represents a novel species within the genus Microlunatus, for which the name Microlunatus panaciterrae sp. nov. is proposed. The type strain is Gsoil 954(T) (=KCTC 13058(T)=DSM 18662(T)).

Ponticoccus gilvus gen. nov., sp. nov., a novel member of the family Propionibacteriaceae from seawater.

A novel actinobacterium, designated strain MSW-19(T), was isolated from a seawater sample in Republic of Korea. Cells were aerobic, Gram-positive, non-endospore-forming, and non-motile cocci. Colonies were circular, convex, opaque, and vivid yellow in colour. A phylogenetic tree based on 16S rRNA gene sequences exhibited that the organism formed a distinct clade within the radius encompassing representatives of the family Propionibacteriaceae. The phylogenetic neighbors were the type strains of the genera Friedmanniella, Microlunatus, Micropruina, Propionicicella, and Propionicimonas. Levels of 16S rRNA gene sequence similarity between the isolate and members of the family were less than 95.3%. The cell wall peptidoglycan of the organism contained LL-diaminopimelic acid as the diagnostic diamino acid. The isolate contained MK-9(H(4)) as the predominant menaquinone, ai-C(15:0) as the major fatty acid and polar lipids including phosphatidylglycerol, phosphatidylethanolamine, and an unknown phospholipid. The G+C content of the DNA was 69.6 mol%. On the basis of the phenotypic and phylogenetic data presented here, the isolate is considered to represent a novel genus and species in the family Propionibacteriaceae, for which the name Ponticoccus gilvus gen. nov., sp. nov. is proposed. The type strain is strain MSW-19(T) (= KCTC 19476(T)= DSM 21351(T)).

Actinopolymorpha rutila sp. nov., isolated from a forest soil.

A Gram-positive, aerobic actinomycete, designated strain YIM 45725(T), was isolated from a forest soil sample in Yunnan Province, China. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate was a member of the family Nocardioidaceae and was related most closely to Actinopolymorpha singaporensis IM 7744(T) (99.4 % 16S rRNA gene sequence similarity). Chemotaxonomic data, including cell-wall components, menaquinones and polar lipids, supported the affiliation of strain YIM 45725(T) to the genus Actinopolymorpha. The results of physiological and biochemical investigations and DNA-DNA hybridization experiments demonstrated that strain YIM 45725(T) differed from the single recognized species of the genus Actinopolymorpha. Strain YIM 45725(T) is therefore considered to represent a novel species of the genus Actinopolymorpha, for which the name Actinopolymorpha rutila sp. nov. is proposed. The type strain is YIM 45725(T) (=CCTCC AA 206004(T)=DSM 18448(T)).

Aestuariimicrobium kwangyangense gen. nov., sp. nov., an LL-diaminopimelic acid-containing bacterium isolated from tidal flat sediment.

Four Gram-positive, catalase-positive, short rod- or coccoid-shaped bacterial strains, R27(T), R44, R45 and R47, were isolated from an enrichment culture with diesel oil-degradation activity and their taxonomic positions were investigated using a polyphasic approach. Phenotypic, phylogenetic and genetic similarities indicated that strains R27(T), R44, R45 and R47 belong to the same species. Phylogenetic analysis based on 16S rRNA gene sequences showed that the four strains form a distinct evolutionary lineage within the family Propionibacteriaceae. The novel four strains had cell-wall peptidoglycan based on LL-diaminopimelic acid, MK-9(H(4)) as the predominant menaquinone and anteiso-C(15 : 0) as the major cellular fatty acid. The DNA G+C contents were 68.8-69.2 mol%. These chemotaxonomic properties, together with phylogenetic distinctiveness, distinguish the four novel strains from recognized members of the family Propionibacteriaceae. On the basis of phenotypic, chemotaxonomic, phylogenetic and genetic data, strains R27(T), R44, R45 and R47 are classified as representatives of a new genus and novel species, Aestuariimicrobium kwangyangense gen. nov., sp. nov., within the family Propionibacteriaceae. The type strain of Aestuariimicrobium kwangyangense sp. nov. is R27(T) (=KCTC 19182(T)=JCM 14204(T)).

Nocardioides alkalitolerans sp. nov., isolated from an alkaline serpentinite soil in Korea.

Four Gram-positive, rod- or coccus-shaped bacterial strains, KSL-1(T), KSL-9, KSL-10 and KSL-12, were isolated from an alkaline serpentinite soil in Korea, and their taxonomic positions were investigated in a polyphasic study. The four strains exhibited no difference in their 16S rRNA gene sequences. Phylogenetic analyses based on 16S rRNA gene sequences showed that the four strains were phylogenetically affiliated to the genus Nocardioides. The four strains had cell-wall peptidoglycan based on ll-diaminopimelic acid as the diamino acid, indicating wall chemotype I. The predominant menaquinone detected in the four strains was MK-8(H(4)). The major fatty acid components were iso-C(16 : 0), 10-methyl-C(18 : 0), C(18 : 1)omega9c and C(17 : 1)omega6c. The DNA G+C contents were 72.4-73.6 mol%. The four strains exhibited 16S rRNA gene sequence similarity levels of 94.0-96.3 % to the type strains of Nocardioides species with validly published names. DNA-DNA relatedness levels between the four strains were 85-91 %. On the basis of phenotypic properties, phylogenetic distinctiveness and genotypic relatedness, strains KSL-1(T), KSL-9, KSL-10 and KSL-12 were classified in the genus Nocardioides as members of a novel species, Nocardioides alkalitolerans sp. nov. The type strain is strain KSL-1(T) (=KCTC 19037(T)=DSM 16699(T)).

Propionimicrobium gen. nov., a new genus to accommodate Propionibacterium lymphophilum (Torrey 1916) Johnson and Cummins 1972, 1057AL as Propionimicrobium lymphophilum comb. nov.

Based upon significant differences in chemotaxonomic properties, i.e., amino acid composition of peptidoglycan, fatty acids and base composition of DNA, and supported by the phylogenetic position of the 165 rDNA sequence the species Propionibacterium lymphophilum was reclassified as Propionimicrobium lymphophilum comb. nov.

Friedmanniella spumicola sp. nov. and Friedmanniella capsulata sp. nov. from activated sludge foam: gram-positive cocci that grow in aggregates of repeating groups of cocci.

Two Gram-positive, non-motile, non-spore-forming, strictly aerobic, pigmented cocci, strains Ben 107T and Ben 108T, growing in aggregates were isolated from activated sludge samples by micromanipulation. Both possessed the rare type A3 gamma' peptidoglycan. Major menaquinones of strain Ben 107T were MK-9(H4) and MK-7(H2), and the main cellular fatty acid was 12-methyltetradecanoic acid (ai-C15:0). In strain Ben 108T, MK-9(H4), MK-9(H2) and MK-7(H4) were the menaquinones and again the main fatty acid was 12-methyltetradecanoic acid (ai-C15:0). Polar lipids in both strains consisted of phosphatidyl inositol, phosphatidyl glycerol and diphosphatidyl glycerol with two other unidentified glycolipids and phospholipids also present in both. These data, together with the 16S rDNA sequence data, suggest that strain Ben 107T belongs to the genus Friedmanniella which presently includes a single recently described species, Friedmanniella antarctica. Although the taxonomic status of strain Ben 108T is far less certain, on the basis of its 16S rRNA sequence it is also adjudged to be best placed in the genus Friedmanniella. The chemotaxonomic characteristics and DNA-DNA hybridization data support the view that Ben 107T and Ben 108T are novel species of the genus Friedmanniella. Hence, it is proposed that strain Ben 107T (= ACM 5121T) is named as Friedmanniella spumicola sp. nov. and strain Ben 108T (= ACM 5120T) as Friedmanniella capsulata sp. nov.

Nocardioides pyridinolyticus sp. nov., a pyridine-degrading bacterium isolated from the oxic zone of an oil shale column.

A bacterial strain which is able to degrade pyridine was previously isolated from the oxic zone of an oil shale column and described as Pimelobacter sp. strain OS4T. However, Pimelobacter species have been transferred to the genera Nocardioides and Terrabacter. Strain OS4T was identified as a member of the genus Nocardioides on the basis of chemotaxonomic analysis and phylogenetic inference based on 16S ribosomal DNA (rDNA) sequence analysis. The G+C content of strain OS4T is 72.5 mol%. The cell wall peptidoglycan contains LL-diaminopimelic acid as the diamino acid. The predominant menaquinone is MK-8(H4). The cellular fatty acid profile of strain OS4T is similar to that of the genus Nocardioides. The 16S rDNA similarity of strain OS4T with previously described Nocardioides species is 94.5% +/- 0.7%, and a phylogenetic tree based on 16S rDNA sequences revealed a distinct lineage for strain OS4T within the evolutionary radiation enclosed by the genus Nocardioides. Therefore, on the basis of our data, we propose that strain OS4T should be placed in the genus Nocardioides as a member of a new species, Nocardioides pyridinolyticus. The type strain of the new species is strain OS4 (= KCTC 0074BP).

Description of Nocardiopsis synnemataformans sp. nov., elevation of Nocardiopsis alba subsp. prasina to Nocardiopsis prasina comb. nov., and designation of Nocardiopsis antarctica and Nocardiopsis alborubida as later subjective synonyms of Nocardiopsis dassonvillei.

Data from chemotaxonomic and 16S ribosomal DNA sequence analyses of an isolate obtained from the sputum of a kidney transplant patient identified the isolate as a member of the genus Nocardiopsis. DNA-DNA hybridization data, as well as physiological characteristics, indicated that the isolate represents a new species of the genus Nocardiopsis, designated Nocardiopsis synnemataformans; the type strain is strain IMMIB D-1215 (= DSM 44143). In addition, DNA-DNA hybridization data, as well as the results of biochemical tests, indicated that Nocardiopsis alborubida DSM 40465T, Nocardiopsis antarctica DSM 43884T, and Nocardiopsis dassonvillei DSM 43111T represent a single species designated N. dassonvillei. We also found that Nocardiopsis alba subsp. alba DSM 43377T and N. alba subsp. prasina DSM 43845T are genetically different and therefore propose that N. alba subsp. prasina be elevated to species status as Nocardiopsis prasina comb. nov., whose type strain is strain DSM 43845.