Cutibacterium acnes-derived extracellular vesicles promote tumor growth in renal cell carcinoma.

Bacterial flora are present in various parts of the human body, including the intestine, and are thought to be involved in the etiology of various diseases such as multiple sclerosis, intestinal diseases, cancer, and uterine diseases. In recent years, the presence of bacterial 16S rRNA genes has been revealed in blood, which was previously thought to be a sterile environment, and characteristic blood microbiomes have been detected in various diseases. However, the mechanism and the origin of the bacterial information are unknown. In this study, we performed 16S rRNA metagenomic analysis of bacterial DNA in serum extracellular vesicles from five healthy donors and seven patients with renal cell carcinoma and detected Cutibacterium acnes DNA as a characteristic bacterial DNA in the serum extracellular vesicles of patients with renal cell carcinoma. In addition, C. acnes DNA was significantly reduced in postoperative serum extracellular vesicles from patients with renal cell carcinoma compared with that in preoperative serum extracellular vesicles from these patients and was also detected in tumor tissue and extracellular vesicles from tumor tissue-associated microbiota, suggesting an association between C. acnes extracellular vesicles and renal cell carcinoma. C. acnes extracellular vesicles were taken up by renal carcinoma cells to enhance their proliferative potential. C. acnes extracellular vesicles also exhibited tumor-promoting activity in a mouse model of renal cancer allografts with enhanced angiogenesis. These results suggest that extracellular vesicles released by C. acnes localized in renal cell carcinoma tissues act in a tumor-promoting manner.

Cutibacterium acnes biofilm formation is influenced by bone microenvironment, implant surfaces and bacterial internalization.

BACKGROUND: The bacterial persistence, responsible for therapeutic failures, can arise from the biofilm formation, which possesses a high tolerance to antibiotics. This threat often occurs when a bone and joint infection is diagnosed after a prosthesis implantation. Understanding the biofilm mechanism is pivotal to enhance prosthesis joint infection (PJI) treatment and prevention. However, little is known on the characteristics of Cutibacterium acnes biofilm formation, whereas this species is frequently involved in prosthesis infections. METHODS: In this study, we compared the biofilm formation of C. acnes PJI-related strains and non-PJI-related strains on plastic support and textured titanium alloy by (i) counting adherent and viable bacteria, (ii) confocal scanning electronic microscopy observations after biofilm matrix labeling and (iii) RT-qPCR experiments. RESULTS: We highlighted material- and strain-dependent modifications of C. acnes biofilm. Non-PJI-related strains formed aggregates on both types of support but with different matrix compositions. While the proportion of polysaccharides signal was higher on plastic, the proportions of polysaccharides and proteins signals were more similar on titanium. The changes in biofilm composition for PJI-related strains was less noticeable. For all tested strains, biofilm formation-related genes were more expressed in biofilm formed on plastic that one formed on titanium. Moreover, the impact of C. acnes internalization in osteoblasts prior to biofilm development was also investigated. After internalization, one of the non-PJI-related strains biofilm characteristics were affected: (i) a lower quantity of adhered bacteria (80.3-fold decrease), (ii) an increase of polysaccharides signal in biofilm and (iii) an activation of biofilm gene expressions on textured titanium disk. CONCLUSION: Taken together, these results evidenced the versatility of C. acnes biofilm, depending on the support used, the bone environment and the strain.

MALDI-TOF mass spectrometry misidentification of Cutibacterium namnetense and Cutibacterium modestum: Implications for multiplex PCR phylotyping of Cutibacterium acnes.

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can misidentify Cutibacterium namnetense and Cutibacterium modestum as Cutibacterium acnes. We now describe how such MALDI-TOF MS misidentification explains previous reports of C. acnes isolates that could not be characterised using a multiplex PCR phylotyping assay.

A New Pathway for Forming Acetate and Synthesizing ATP during Fermentation in Bacteria.

Many bacteria and other organisms carry out fermentations forming acetate. These fermentations have broad importance for foods, agriculture, and industry. They also are important for bacteria themselves because they often generate ATP. Here, we found a biochemical pathway for forming acetate and synthesizing ATP that was unknown in fermentative bacteria. We found that the bacterium Cutibacterium granulosum formed acetate during fermentation of glucose. It did not use phosphotransacetylase or acetate kinase, enzymes found in nearly all acetate-forming bacteria. Instead, it used a pathway involving two different enzymes. The first enzyme, succinyl coenzyme A (succinyl-CoA):acetate CoA-transferase (SCACT), forms acetate from acetyl-CoA. The second enzyme, succinyl-CoA synthetase (SCS), synthesizes ATP. We identified the genes encoding these enzymes, and they were homologs of SCACT and SCS genes found in other bacteria. The pathway resembles one described in eukaryotes, but it uses bacterial, not eukaryotic, gene homologs. To find other instances of the pathway, we analyzed sequences of all biochemically characterized homologs of SCACT and SCS (103 enzymes from 64 publications). Homologs with similar enzymatic activity had similar sequences, enabling a large-scale search for them in genomes. We searched nearly 600 genomes of bacteria known to form acetate, and we found that 6% encoded homologs with SCACT and SCS activity. This included >30 species belonging to 5 different phyla, showing that a diverse range of bacteria encode the SCACT/SCS pathway. This work suggests the SCACT/SCS pathway is important for acetate formation in many branches of the tree of life. IMPORTANCE Pathways for forming acetate during fermentation have been studied for over 80 years. In that time, several pathways in a range of organisms, from bacteria to animals, have been described. However, one pathway (involving succinyl-CoA:acetate CoA-transferase and succinyl-CoA synthetase) has not been reported in prokaryotes. Here, we discovered enzymes for this pathway in the fermentative bacterium Cutibacterium granulosum. We also found >30 other fermentative bacteria that encode this pathway, demonstrating that it could be common. This pathway represents a new way for bacteria to form acetate from acetyl-CoA and synthesize ATP via substrate-level phosphorylation. It could be a target for controlling yield of acetate during fermentation, with relevance for foods, agriculture, and industry.

Aestuariimicrobium ganziense sp. nov., a new Gram-positive bacterium isolated from soil in the Ganzi Tibetan autonomous prefecture, China.

A novel Gram-stain positive, oval-shaped, and non-flagellated bacterium, designated YIM S02566T, was isolated from alpine soil in Shadui Towns, Ganzi County, Ganzi Tibetan Autonomous Prefecture, Sichuan Province, PR China. Growth occurred at 23-35 °C (optimum, 30 °C) in the presence of 0.5-4% (w/v) NaCl (optimum, 1%) and at pH 7.0-8.0 (optimum, pH 7.0). The phylogenetic analysis based on 16S rRNA gene sequence revealed that strain YIM S02566T was most closely related to the genus Aestuariimicrobium, with Aestuariimicrobium kwangyangense R27T and Aestuariimicrobium soli D6T as its closest relative (sequence similarities were 96.3% and 95.4%, respectively). YIM S02566T contained LL-diaminopimelic acid in the cell wall. MK-9(H4) was the predominant menaquinone. The major fatty acid patterns were anteiso-C15:0 (60.0%). The major polar lipid was DPG. The genome size of strain YIM S02566T was 3.1 Mb, comprising 3078 predicted genes with a DNA G + C content of 69.0 mol%. Based on these genotypic, chemotaxonomic and phenotypic evidences, strain YIM S02566T was identified as a novel species in the genus Aestuariimicrobium, for which the name Aestuariimicrobium ganziense sp. nov. is proposed. The type strain is YIM S02566T (= CGMCC 1.18751 T = KCTC 49,477 T).

Unravelling the eco-specificity and pathophysiological properties of Cutibacterium species in the light of recent taxonomic changes.

In 2016, a new species name Cutibacterium acnes was coined for the well-documented species, Propionibacterium acnes, one of the most successful and clinically important skin commensals. The nomenclatural changes were brought about through creation of the genus Cutibacterium, when a group of propionibacteria isolates from the skin were transferred from the genus Propionibacterium and placed in the phylum Actinobacteria. Almost simultaneously, the discovery of two novel species of Cutibacterium occurred and the proposal of three subspecies of C. acnes were reported. These dramatic changes that occurred in a long-established taxon made it challenging for the non-specialist to correlate the huge volume of hitherto published work with current findings. In this review, we aim to correlate the eco-specificity and pathophysiological properties of these newly circumscribed taxa. We envisage that this information will shed light on the pathogenic potential of new isolates and enable better assessment of their clinical importance in the foreseeable future. Currently, five species are recognized within the genus: Cutibacterium acnes, Cutibacterium avidum, Cutibacterium granulosum, Cutibacterium modestum (previously, "Propionibacterium humerusii"), and Cutibacterium namnetense. These reside in different niches reflecting their uniqueness in their genetic makeup. Their pathogenicity includes acne inflammation, sarcoidosis, progressive macular hypomelanosis, prostate cancer, and infections (bone, lumbar disc, and heart). This is also the case for the three newly described subspecies of C. acnes, which are C. acnes subspecies acnes (C. acnes type I), subspecies defendens (C. acnes type II), and subspecies elongatum (C. acnes type III). C. acnes subspecies acnes is related to inflamed acne and sarcoidosis, while subspecies defendens to prostate cancer and subspecies elongatum to progressive macular hypomelanosis. Because the current nomenclature is based upon polyphasic analyses of the biochemical and pathogenic characteristics and comparative genomics, it provides a sound basis studying the pathophysiological roles of these species.

Bacterial pericarditis and empyema caused by Cutibacterium acnes in a patient with metastatic lung cancer.

Bacterial pericarditis and empyema due to Cutibacterium acnes has rarely been reported. C.acnes, a normal component of human skin flora, is often considered a contaminant when isolated from body fluids and thus cases may be underreported. We report the first case of concurrent purulent pericarditis and empyema caused by C. acnes in a patient with newly diagnosed metastatic lung cancer. Our patient underwent pericardial window creation and placement of pericardial and bilateral chest tubes and was successfully treated with culture directed antibiotic therapy.

Nisin J, a Novel Natural Nisin Variant, Is Produced by Staphylococcus capitis Sourced from the Human Skin Microbiota.

The skin microbiota is thought to play a key role in host protection from infection. Nisin J is a novel nisin variant produced by Staphylococcus capitis APC 2923, a strain isolated from the toe web space area in a screening study performed on the human skin microbiota. Whole-genome sequencing and mass spectrometry of the purified peptide confirmed that S. capitis APC 2923 produces a 3,458-Da bacteriocin, designated nisin J, which exhibited antimicrobial activity against a range of Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus (MRSA) and Cutibacterium acnes The gene order in the nisin J gene cluster (nsjFEGBTCJP) differs from that of other nisin variants in that it is lacking the nisin regulatory genes, nisRK, as well as the nisin immunity gene nisI Nisin J has 9 amino acid changes compared to prototypical nisin A, with 8 amino acid substitutions, 6 of which are not present in other nisin variants (Ile4Lys, Met17Gln, Gly18Thr, Asn20Phe, Met21Ala, Ile30Gly, Val33His, and Lys34Thr), and an extra amino acid close to the C terminus, rendering nisin J the only nisin variant to contain 35 amino acids. This is the first report of a nisin variant produced by a Staphylococcus species and the first nisin producer isolated from human skin.IMPORTANCE This study describes the characterization of nisin J, the first example of a natural nisin variant, produced by a human skin isolate of staphylococcal origin. Nisin J displays inhibitory activity against a wide range of bacterial targets, including MRSA. This work demonstrates the potential of human commensals as a source for novel antimicrobials that could form part of the solution to antibiotic resistance across a broad range of bacterial pathogens.

Transferable Multidrug-Resistance Plasmid Carrying a Novel Macrolide-Clindamycin Resistance Gene, erm(50), in Cutibacterium acnes.

Antimicrobial-resistant Cutibacterium acnes strains have emerged and disseminated throughout the world. The 23S rRNA mutation and erm(X) gene are known as the major resistance determinants of macrolides and clindamycin in C. acnes We isolated eight high-level macrolide-clindamycin-resistant C. acnes strains with no known resistance determinants, such as 23S rRNA mutation and erm(X), from different acne patients in 2008 between 2013 and 2015. The aim of this study was to identify the novel mechanisms of resistance in C. acnes Whole-genome sequencing revealed the existence of a plasmid DNA, denoted pTZC1 (length, 31,440 bp), carrying the novel macrolide-clindamycin resistance gene erm(50) and tetracycline resistance gene tet(W). pTZC1 was detected in all C. acnes isolates (eight strains) exhibiting high-level macrolide-clindamycin resistance, with no known resistance determinants (MIC of clarithromycin, ≥256 µg/ml; clindamycin, ≥256 µg/ml). Transconjugation experiments demonstrated that the pTZC1 was horizontally transferred among C. acnes strains and conferred resistance to macrolides, clindamycin, and tetracyclines. Our data showed, for the first time, the existence of a transferable multidrug-resistant plasmid in C. acnes Increased prevalence of this plasmid will be a great threat to antimicrobial therapy for acne vulgaris.

Cutibacterium acnes Isolates from Deep Tissue Specimens Retrieved during Revision Shoulder Arthroplasty: Similar Colony Morphology Does Not Indicate Clonality.

Cutibacterium acnes is the most common bacterium associated with periprosthetic shoulder infections. Sequencing of C. acnes has been proposed as a potential rapid diagnostic tool and a method of determining subtypes associated with pathogenicity and antibiotic resistance patterns. When multiple deep samples from the same surgery are culture positive for the same species and the isolates show the same culture phenotype, it is typically assumed that these isolates are clonal. However, it is well-known that C. acnes is not clonal on the skin of most individuals. We hypothesized that the C. acnes bacteria recovered at the time of revision shoulder arthroplasty would often represent more than one subtype, and we tested this hypothesis in this work. For patients undergoing revision shoulder arthroplasty, multiple samples from the surgical field were taken. For those patients with multiple samples that were culture positive for C. acnes, isolates from each sample were subjected to full genome sequencing. Of 11 patients, 5 (45%) had different subtypes of C. acnes within the deep tissues even though the colony morphology was similar. One patient had four subtypes in the deep tissues, while four patients had two different subtypes. Up to four different subtypes of C. acnes were observed in the deep tissues of a single patient. Clonality of C. acnes isolates from deep specimens from a potential periprosthetic shoulder infection cannot be assumed. Sequence-based characterization of virulence and antibiotic resistance may require testing of multiple deep specimens.

Microlunatus speluncae sp. nov., a novel actinobacterium isolated from a Karstic subterranean environment sample.

A novel actinobacterial strain, designated SYSU K12189T, was isolated from a soil sample collected from a Karst cave in Xingyi county, Guizhou province, south-western China. The taxonomic position of the strain was investigated using a polyphasic approach. Cells of the strain were observed to be aerobic and Gram-stain positive. On the basis of 16S rRNA gene sequence similarities and phylogenetic analysis, strain SYSU K12189T is closely related to the type strains of the genus Microlunatus, Microlunatus parietis 12-Be-011T (98.5% sequence similarity), Microlunatus nigridraconis CPCC 203993T (98.4%) and Microlunatus cavernae YIM C01117T (96.6%), and is therefore considered to represent a member of the genus Microlunatus. DNA-DNA hybridization values between strain SYSU K12189T and related type strains of the genus Microlunatus were < 70%. In addition, LL-diaminopimelic acid was found to be the diagnostic diamino acid in the cell wall peptidoglycan. The major isoprenoid quinone was identified as MK-9(H4), while the major fatty acids (> 10%) were found to be anteiso-C15:0, iso-C15:0, iso-C16:0 and iso-C14:0. The polar lipids were found to contain diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, three glycolipids and two unidentified lipids. The genomic DNA G+C content of strain SYSU K12189T was determined to be 69.4 mol%. On the basis of phenotypic, genotypic and phylogenetic data, strain SYSU K12189T is concluded to represent a novel species of the genus Microlunatus, for which the name Microlunatus speluncae sp. nov. is proposed. The type strain is SYSU K12189T (= KCTC 39847T = DSM 103947T).

Microbiological Profile of Sarecycline, a Novel Targeted Spectrum Tetracycline for the Treatment of Acne Vulgaris.

Sarecycline is the first narrow-spectrum tetracycline-class antibiotic being developed for acne treatment. In addition to exhibiting activity against important skin/soft tissue pathogens, sarecycline exhibits targeted antibacterial activity against clinical isolates of Cutibacterium acnes In the current study, sarecycline was 16- to 32-fold less active than broad-spectrum tetracyclines-such as minocycline and doxycycline-against aerobic Gram-negative bacilli associated with the normal human intestinal microbiome. Also, reduced activity against Escherichia coli was observed in vivo in a murine septicemia model, with the 50% protective doses, or the doses required to achieve 50% survival, being >40 mg/kg of body weight and 5.72 mg/kg for sarecycline and doxycycline, respectively. Sarecycline was also 4- to 8-fold less active than doxycycline against representative anaerobic bacteria that also comprise the normal human intestinal microbiome. Additionally, C. acnes strains displayed a low propensity for the development of resistance to sarecycline, with spontaneous mutation frequencies being 10-10 at 4 to 8 times the MIC, similar to those for minocycline and vancomycin. When tested against Gram-positive pathogens with defined tetracycline resistance mechanisms, sarecycline was more active than tetracycline against tet(K) and tet(M) strains, with MICs ranging from 0.125 to 1.0 µl/ml and 8 µl/ml, respectively, compared with MICs of 16 to 64 µl/ml and 64 µl/ml for tetracycline, respectively. However, sarecycline activity against the tet(K) and tet(M) strains was decreased compared to that against the wild type, which demonstrated MICs ranging from 0.06 to 0.25 µl/ml, though the decrease in the activity of sarecycline against the tet(K) and tet(M) strains was not as pronounced as that of tetracycline. These findings support sarecycline as a narrow-spectrum tetracycline-class antibiotic that is effective for the treatment of acne, and further investigation into the potential reduced effects on the gut microbiome compared with those of other agents is warranted.

Classification of Cutibacterium acnes at phylotype level by MALDI-MS proteotyping.

Cutibacterium acnes is a major commensal human skin bacteria. It is a producer of propionic acids that maintain skin acidic pH to inhibit the growth of pathogens. On the other hand, it is also associated with diseases such as acne vulgaris and sarcoidosis. C. acnes strains have been classified into six phylotypes using DNA-based approaches. Because several characteristic features of C. acnes vary according to the phylotype, the development of a practical method to identify these phylotypes is needed. For rapid identification of phylotypes for C. acnes strains, a matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) fingerprinting technique has been applied; however, some phylotypes have not been discriminated. We developed a high-throughput protein purification method to detect biomarker proteins by ultrafiltration. MALDI-MS proteotyping using profiling of identified biomarker peaks was applied for the classification of 24 strains of C. acnes, and these were successfully classified into the correct phylotypes. This is a promising method that allows the discrimination of C. acnes phylotypes independent of a DNA-based approach.

Intra-peritoneal abscess after an abdominal hysterectomy involving Cutibacterium avidum (former Propionibacterium avidum) highly resistant to clindamycin.

Cutibacterium avidum is a gram-positive anaerobic rod belonging to the cutaneous group of human bacteria with preferential colonization of sweat glands in moist areas. The microorganism rarely cause disease, generally delayed prosthetic joint infections (PJIs). We describe the second case of intraperitoneal abscess by C. avidum after an abdominal surgery in an obese female patient and the first case after a non-prosthetic abdominal surgery due to a highly clindamycin resistant strain in a patient with underling conditions. The patient was successfully treated with surgical drainage and beta-lactam antibiotics. Although rare and apparently non-pathogenic, C. avidum may be involved in infections, especially in some high-risk patients with obesity who have undergone surgical incision involving deep folder of the skin. The microorganism was identified by phenotypic methods, MALDI-TOF MS and 16S rRNA gene sequencing. Susceptibility test should be performed in C. avidum because high level resistance to clindamycin could be present. We present a literature review of C. avidum infections.

Aestuariimicrobium soli sp. nov., isolated from farmland soil, and emended description of the genus Aestuariimicrobium.

A Gram-reaction-positive, catalase-positive, non-spore-forming and short rod- or oval-shaped bacterial strain, designated D6T, was isolated from farmland soil in Xuancheng, Anhui Province, China. Growth occurred at 4-37 °C (optimum, 30 °C), at pH 6.5-8.5 (optimum, 7.0) and with 0-7 % (w/v) NaCl (optimum, 0.5 % NaCl). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain D6T was most closely related to Aestuariimicrobium kwangyangense DSM 21549T (98.47 %), followed by Tessaracoccus rhinocerotis YIM 101269T (94.46 %). Strain D6T had a cell-wall peptidoglycan based on ll-diaminopimelic acid. MK-9(H4) was the predominant menaquinone. The major fatty acids of strain D6T were anteiso-C15 : 0, iso-C15 : 0 and summed feature 4 (iso-C17 : 1 I and/or anteiso-C17 : 1 B). The major polar lipids were a lipid, glycolipid and phospholipid. The DNA G+C content was 69.2 mol% and strain D6T showed low DNA-DNA relatedness to A. kwangyangense DSM 21549T (36.45±0.42 %). Based on these genotypic and phenotypic data, strain D6T represents a novel species in the genus Aestuariimicrobium, for which the name Aestuariimicrobium soli sp. nov. is proposed. The type strain is D6T (=KCTC 39995T=DSM 105824T). An emended description of the genus Aestuariimicrobium is presented.

Tessaracoccus aquimaris sp. nov., isolated from the intestine of a Korean rockfish, Sebastes schlegelii, from a marine aquaculture pond.

A novel coccus-shaped, Gram-stain-positive, non-motile and aerobic bacterium, designated strain NSG39T, was isolated from the intestine of a Korean rockfish, Sebastes schlegelii. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the newly isolated strain NSG39T was closely related to Tessaracoccus flavus RP1T (98.0 %). The isolate grew at 15-37 °C, pH 7-9 and 0-4 % (w/v) salinity, with optimal growth at 30 °C, pH 8 and 0 % (w/v) salinity. The cell wall of the organism contained ll-diaminopimelic acid as a diagnostic diamino acid, and ribose, mannose, glucose and galactose as diagnostic sugars. The polar lipid comprised diphosphatidylglycerol, phosphatidylglycerol, three glycolipids and four unidentified polar lipids. The major cellular fatty acid was anteiso-C15 : 0 (47.2 %). The major menaquinone was MK-9 (H4). The DNA G+C content of the isolate was 68.8 mol%. The genome-based orthologous average nucleotide identity value for strain NSG39T and T. flavus RP1T was 76.6 %. Based on the phylogenetic analysis and its biological characteristics, strain NSG39T is considered to represent a novel species of the genus Tessaracoccus, for which the name Tessaracoccus aquimaris is proposed. The type strain is NSG39T (=KACC 17540T=JCM 19289T).

Tessaracoccus terricola sp. nov., isolated from oil-contaminated soil.

A Gram-stain-positive, non-motile, yellow and rod-shaped actinobacterium, designated strain Brt-AT, isolated from oil-contaminated soil, grew at 15-40 °C, at pH 5.5-10.0 and at 0-2 % (w/v) NaCl concentration. This strain was characterized by a polyphasic approach. The 16S rRNA gene sequence analysis showed that strain Brt-AT belonged to the genus Tessaracoccus and is closely related to Tessaracoccus rhinocerotis YIM 101269T, Tessaracoccus flavescens SST-39T, Tessaracoccus defluvii LNB-140T and Tessaracoccus flavus RP1T (99.03, 97.00, 96.88, and 96.46 % gene sequence similarity, respectively). The predominant respiratory quinone was MK-9(H4); the major polar lipids were phosphatidylglycerol and diphosphatidylglycerol; the predominant polyamines were spermine and spermidine; and the major fatty acids were anteiso-C15 : 0 and iso-C15 : 0. The cell-wall peptidoglycan contained ll-diaminopimelic acid; and glucose and ribose were detected as diagnostic sugars from whole-cell hydrolysates. The DNA G+C content was 68.1 mol%. The DNA-DNA relatedness between strain Brt-AT and its closely related species of the genus Tessaracoccus were between 55.0-44.0 %, which fall below the threshold value of 70 % for the strain to be considered as novel. The morphological, physiological, chemotaxonomic and phylogenetic analyses clearly distinguished this strain from its closest phylogenetic neighbours. Thus, strain Brt-AT represents a novel species of the genus Tessaracoccus, for which the name Tessaracoccus terricola sp. nov. is proposed. The type strain is Brt-AT (=KEMB 9005-690T=KACC 19391T=JCM 32157T).

Pseudopropionibacterium rubrum sp. nov., a novel red-pigmented species isolated from human gingival sulcus.

In this study, Strain [corrected] SK-1(T), a novel gram-positive, pleomorphic, rod-shaped, non-spore forming, non-motile organism, designated SK-1T , was isolated from human gingival sulcus and found to produce acetic acid, propionic acid, lactic acid, and succinic acid as end products of glucose fermentation. Strain SK-1T is most closely related to Pseudopropionibacterium (Propionibacterium) propionicum with sequence homologies of the 16S rRNA and RNA polymerase ß subunit (rpoB) genes of 96.6% and 93.1%, respectively. The genomic DNA G + C content of the isolate was 61.8 mol%. On the basis of the sequence data of the 16S rRNA and housekeeping (rpoB) genes, a novel taxon is here proposed, Pseudopropionibacterium rubrum sp. nov. (type strain SK-1T = JCM 31317T = DSM 100122T ). The 16S rRNA and rpoB gene sequences of strain SK-1T have been deposited in the DDBJ under the accession numbers LC002971 and LC102236, respectively.

Kribbella sindirgiensis sp. nov. isolated from soil.

A Kribbella strain FSN23T was isolated from soil sample which was collected from Caygoren Dam lakeside located in Sindirgi, Turkey. The isolate was investigated using a polyphasic approach consisting of numeric, chemotaxonomic and molecular analysis. The isolate indicated chemotaxonomic, morphological and phylogenetic properties associated with members of the genus Kribbella. Phylogenetic analysis based on the 16S rRNA sequence of the strain demonstrated that the strain forms a subclade with K. aluminosa HKI 0478T and K. jejuensis HD9T. The organism formed an extensively branched substrate and aerial hyphae which generated spiral chains of spores with smooth surfaces. The cell wall contained LL-diaminopimelic acid, and the whole cell sugars were glucose and ribose along with trace amounts of mannose. The polar lipids were identified as phosphatidylglycerol, diphosphatidylglycerol, four unidentified lipids and five unidentified polar lipids. The predominant menaquinone was MK-9(H4). The major cellular fatty acids were anteiso-C15:0 and iso-C16:0. Polyphasic taxonomy properties confirm that strain FSN23T represents a novel Kribbella taxon distinguished from closely related type strains. Hence, strain FSN23T (=KCTC 29220T = DSM 27082T) is proposed as the type strain of a novel species with the name Kribbella sindirgiensis sp. nov.

Description of Naumannella cuiyingiana sp. nov., isolated from a ca. 1500-year-old mural painting, and emended description of the genus Naumannella.

Strain AFT2T was isolated from a mural painting sample from a ca. 1500-year-old tomb located in Shanxi Province, China. The isolate was a Gram-stain-positive, non-motile, non-spore-forming, aerobic and oval to short-rod-shaped bacterium that formed white-pigmented colonies. Phylogenetic analyses based on 16S rRNA gene sequence revealed that strain AFT2T was most closely (97.01 %) correlated and formed a monophyletic clade with Naumannella halotolerans WS4616T (=DSM 24323T). The G+C content of the genomic DNA was 71.97 mol%, and the strain showed 37.27 % DNA-DNA relatedness to N. halotolerans DSM 24323T. The major cellular fatty acid was anteiso-C15 : 0 (55.32 %), and MK-9(H4) was the only respiratory quinone. The polar lipids comprised phosphatidylglycerol, diphosphatidylglycerol, two unknown phospholipids and five unknown glycolipids. ll-Diaminopimelic acid was detected in the cell-wall peptidoglycan (type A3γ), and the whole-cell sugars consisted of ribose, mannose, arabinose and galactose. On the basis of its phenotypic and phylogenetic characteristics, it is proposed that strain AFT2T should be classified as a representative of a novel species of the genus Naumannella, for which the name Naumannella cuiyingiana sp. nov. is proposed. The type strain is AFT2T (=CCTCC AB 2015428T=DSM 103164T).