Metabolic Dysfunction-Associated Fatty Liver Disease on Distinct Microbial Communities at the Bacterial Phylum Level.

INTRODUCTION: Limited data are available on the correlation between microbial communities and metabolic dysfunction-associated fatty liver disease (MAFLD). This study aimed to evaluate the influence of MAFLD on diverse microbial communities. METHODS: We recruited 43 patients with a nonviral liver disease. Enrolled patients were divided into two groups according to MAFLD criteria. The fecal microbial composition was evaluated using the variable V3-V4 region of the 16S ribosomal RNA region, which was amplified using polymerase chain reaction. First, we assessed the influence of MAFLD on distinct microbial communities at the bacterial phylum level. Next, the correlation between the microbial communities and diversity in patients with MAFLD was evaluated. RESULTS: Among the enrolled participants, the non-MAFLD and MAFLD groups consisted of 21 and 22 patients, respectively. Sequences were distributed among ten bacterial phyla. The relative abundance of Firmicutes was significantly higher in the MAFLD group than in the non-MAFLD group (p = 0.014). The microbial diversity was not significantly influenced by the presence of MAFLD (Chao-1 index: p = 0.215 and Shannon index: p = 0.174, respectively); nonetheless, the correlation coefficient between the abundances of Firmicutes and microbial diversity was higher in the non-MAFLD group than in the MAFLD group. CONCLUSION: The presence of MAFLD increased the relative abundances of Firmicutes at the bacterial phylum level, which may cause the discrepancy between the abundances of Firmicutes and diversity in patients with MAFLD.

Rhizohabitans arisaemae gen. nov., sp. nov., a novel actinomycete of the family Streptosporangiaceae.

An actinomycete strain K14-0274T was isolated from the root of Arisaema thunbergii Blume subsp. urashima (H. Hara) H. Ohashi et J. Murata collected in Japan. The results of phylogenetic analysis based on the 16S rRNA gene sequence indicated thatK14-0274T could be distinguished from the members of all known genera, although it represented a member of the family Streptosporangiaceae. K14-0274T produced sporangium-like spherical vesicles with spores on white aerial mycelia. MK-9 (H4) and MK-9 (H6) were the major menaquinones. The whole-cell hydrolysates contained madurose, glucose, mannose, rhamnose and ribose. The cell-wall amino acids comprise l-alanine, d-alanine, d-glutamic acid and meso-diaminopimelic acid. The N-acyl type of muramic acid was acetyl. Mycolic acids were not detected. Phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol and phosphatidylinositolmannoside were detected. The predominant fatty acids were iso-C16 : 0, 10-methyl-C18 : 0 and C16 : 0. The G+C content of the genomic DNA was 69.7 mol%. On the basis of morphological, phylogenetic and chemotaxonomic characteristics, strain K14-0427T represents a novel genus in the family Streptosporangiaceae, for which the name Rhizohabitans arisaemae gen. nov., sp. nov. is proposed. The type strain is K14-0247T (=NBRC 114594T =TBRC 12948T).

Amycolatopsis iheyensis sp. nov., isolated from soil on Iheya Island, Japan.

A novel actinomycete, designated strain OK19-0408T, was isolated from soil collected on Iheya island, Okinawa prefecture, Japan. Using the polyphasic taxonomic approach, comparing 16S rRNA gene sequences, the new isolate was found to be most closely related to Amycolatopsis vancoresmycina JCM12675T (98.71 %). Phylogenetic analyses using 16S rRNA sequences indicated that strain OK19-0408T was clustered with Amycolatopsis australiensis JCM15587T. However, digital DNA-DNA hybridization analyses indicated a low relatedness, in the range of 33.9-34.7 %, between strain OK19-0408T and these closely related strains. Strain OK19-0408T contained meso-diaminopimelic acid and whole-cell sugars consisting of arabinose and galactose. The acyl type of the peptidoglycan was acetyl and mycolic acids were absent in strain OK19-0408T. The major menaquinone was MK-9(H4) and hydroxy-phosphatidylethanolamine was detected as the predominant phospholipid. The predominant cellular fatty acid was iso-C16 : 0. The DNA G+C content of the genomic DNA was 71.5 mol%. Based on the polyphasic approach, strain OK19-0408T represents a novel species of the genus Amycolatopsis, for which the name Amycolatopsis iheyensis sp. nov. is proposed. The type strain of the type species is OK19-0408T (=NBRC115671T=TBRC16040T).

Impact of liver fibrosis on the relative abundance of a urease-positive Streptococcus salivarius group from saliva in patients with chronic liver disease.

AIM: We performed genomic analysis to study the relative abundance of a urease-positive Streptococcus salivarius group isolated from the saliva of patients with chronic liver disease. METHODS: Male and female patients with chronic liver disease aged over 20 years were included. First, we assessed the frequency and type of the S. salivarius group isolated from oral saliva using molecular biology techniques based on 16S rRNA and dephospho-coenzyme A kinase gene sequencing. Next, we assessed the correlation between the urease positivity rate in the S. salivarius group isolated from oral saliva and liver fibrosis based on chronic liver disease. Urease-positive strains were identified by the urease test using urea broth (Difco, Franklin Lakes, NJ, USA). Liver fibrosis was evaluated by the liver stiffness measurement value based on magnetic resonance elastography. RESULTS: A total of 45 patients identified using the multiplex polymerase chain reaction for the 16S rRNA gene were tested using the multiplex polymerase chain reaction for the dephospho-coenzyme A kinase gene. Confirming the strains detected in each of the 45 patients, urease-positive S. salivarius was detected in 28 patients (62%), urease-negative S. salivarius in 25 patients (56%), and urease-positive Streptococcus vestibularis in 12 patients (27%). There was no patient with urease-negative S. vestibularis. The urease-positive rate of the S. salivarius group in the cirrhosis and non-cirrhosis groups were 82.2% and 39.2%, respectively. The liver cirrhosis group had a higher urease positivity rate than the non-cirrhotic group (p < 0.001). CONCLUSIONS: Liver fibrosis influences the frequency of a urease-positive S. salivarius group isolated from oral saliva.

Mycelial differentiation linked avermectin production in Streptomyces avermitilis studied with Raman imaging.

Streptomyces avermitilis is a gram-positive bacterium that undergoes complex physiological and morphological differentiation during its life cycle, which has implications in secondary metabolite production. Avermectin, produced by S. avermitilis, is widely used as an anthelmintic and insecticidal agent. In this study, we have applied Raman microspectroscopic imaging to elucidate the correlation between production of avermectin and the morphological differentiation in S. avermitilis. We demonstrate distinctive variations in the localization of secondary metabolites at various stages of morphological differentiation. Under solid culture, avermectin was detected in the mycelia formed at the later stages of morphological differentiation (e.g., spore-bearing mycelium and spiral spore chains), but not in the early-stage substrate mycelium. On the contrary, under liquid culture condition, avermectin was found concentrated in the mycelial pellet formed at the early MII stage of differentiation. Furthermore, the chemical profiles of the mycelia were substantially different depending on the culture condition. Raman spectra corresponding to proteins, lipids, and cytochrome were observed in the mycelia irrespective of the stage of morphological differentiation, however, carotenoid was observed under solid culture condition particularly in spore-bearing mycelium and spiral spore chains. KEY POINTS: • Avermectin production is regulated during mycelial differentiation • Liquid and solid culture conditions affects mycelial differentiation • Raman microspectroscopic analysis reveals localization profiles of avermectin.

The change rate in serum nitric oxide may affect lenvatinib therapy in hepatocellular carcinoma.

BACKGROUND: Lenvatinib is appropriate for reducing the production of nitric oxide (NO) and facilitating as block angiogenesis. However, to our knowledge, there are no data that support the correlation between NO and clinical response in patients who received lenvatinib therapy for HCC. Therefore, we investigated the correlation between the change rate of NO levels and clinical responses including adverse events (AEs) after lenvatinib therapy for unresectable hepatocellular carcinoma (HCC). METHODS: This study was conducted using previously collected data from another study. We enrolled 70 patients who received lenvatinib for advanced or unresectable HCC. NO was measured by converting nitrate (NO3-) to nitrite (NO2-) with nitrate reductase, followed by quantitation of NO2- based on Griess reagent. To determine whether lenvatinib influences NO in unresectable HCC, we evaluated the influence of the change rate of NO from baseline after administration of lenvatinib on maximal therapeutic response and SAE. RESULTS: After lenvatinib administration, a change rate in the NO from 0.27 to 4.16 was observed. There was no difference between the clinical response to lenvatinib and the change rate of NO (p = 0.632). However, the change rate of NO was significantly lower in patients with AEs than in those without AEs (p = 0.030). When a reduction in NO rate of < 0.8 was defined as a clinically significant reduction of NO (CSRN), the CSRN group had significantly worse progression-free survival (PFS) and overall survival (OS) than the non-CSRN group (p = 0.029 and p = 0.005, respectively). CONCLUSION: Decreased NO levels were associated with the occurrence of AEs and worse prognosis after lenvatinib administration. Change rate in serum NO can be used as predictive markers in patients receiving lenvatinib therapy for HCC.

Antibiotic-resistant status and pathogenic clonal complex of canine Streptococcus canis-associated deep pyoderma.

BACKGROUND: Streptococcus canis causes deep pyoderma in canines, which raises concerns about the risk of isolates from lesions acquiring an antibiotic-resistant phenotype. It is necessary to identify effective antibiotics and the characteristics of the pathogenic cluster for S. canis-associated deep pyoderma. RESULTS: The signalment, molecular typing, and antibiotic-resistant status of S. canis isolated from deep pyoderma lesions (27 strains) and oral cavities (26 strains) were analyzed. Older dogs tended to have S. canis-associated deep pyoderma (15 of 27 dogs over 10 years old). Veterinarians chose quinolones for 10/16 cases (63%), even though the rate of quinolone-resistant strains of S. canis is 38-59%. Although 70% of the strains showed resistance to three or more antibiotic classes (37/53), 94% (50/53) strains showed sensitivity for penicillins. We also identified ß-lactamase activity among penicillin-resistant strains of S. canis. Clonal complex 13 (CC13) was detected only in lesions and formed independent clusters in the phylogenetic tree. One strain of CC13 was resistant to the anti-methicillin-resistant Staphylococcus aureus drugs, vancomycin and linezolid. CONCLUSION: Although antibiotic-resistant strains of S. canis are isolated at a high rate, they can currently be treated with ß-lactamase-inhibiting penicillins. CC13 may be a pathogenic cluster with high levels of antibiotics resistance.

Evaluation of Skeletal Muscle Mass in Patients with Chronic Liver Disease Shows Different Results Based on Bioelectric Impedance Analysis and Computed Tomography.

OBJECTIVE: We aimed to evaluate the difference between computed tomography (CT)-based and bioelectrical impedance analysis (BIA)-based assessment of sarcopenia in patients with chronic liver disease (CLD). METHODS: We enrolled a total of 257 patients who were evaluated with or without sarcopenia. Sarcopenia was defined as a low skeletal muscle mass index (SMI) with low muscular strength by the Japan Society of Hepatology. To evaluate whether or not the different methods influence the diagnosis of sarcopenia for patients with CLD, we assessed the number and characteristics of mismatches between the low SMI using BIA and CT. We also compared the overall survival (OS) in patients with and without sarcopenia based on CT and BIA to evaluate the appropriate methods. RESULTS: The numbers of patients with low SMI using BIA or CT were 53 (20.6%) and 114 (44.3%) patients, respectively. Multivariate analysis revealed that hepatic ascites and body weight were independent factors of mismatch between SMI using BIA versus CT (hazard ratio [HR] 3.232, p < 0.001; HR 2.347, p = 0.005, respectively). The median OS in patients with sarcopenia based on CT was significantly lower than that in patients without sarcopenia (p = 0.006). In contrast, there was no difference between patients with sarcopenia based on BIA (p = 0.217). CONCLUSION: Muscle mass in patients with CLD may be overestimated by the BIA method compared to CT when assessing sarcopenia, especially in cases of fluid retention.

Fecal microbiota transplantation as therapy for recurrent Clostridioides difficile infection is associated with amelioration of delirium and accompanied by changes in fecal microbiota and the metabolome.

Recurrent Clostridioides difficile infection (rCDI) is a frustrating condition that may affect a person's quality of life for months. Microbiome-based therapy such as fecal microbiota transplantation (FMT) has been effective for the treatment of rCDI by correcting the imbalance of the gut microbiota. Appropriate antibiotic treatment is recommended for at least two recurrences before offering FMT. Here, we report the case of a 92-year-old woman who experienced five recurrences of Clostridioides difficile infection (CDI) (six episodes in total) complicated by dementia and delirium, both of which were dramatically improved by FMT, which was associated with alterations in fecal microbiota and the metabolome. Analyses of whole microbial communities and metabolomic analyses were performed on stool specimens collected from the patient on the first episode, the third episode, the day of FMT (before FMT), and 2, 8, and 23 weeks after the FMT and from the donor. The patient had various fecal dysbioses on the first and third episodes and on the day of FMT. Two weeks after FMT, diversity of the gut bacteriome as well as the virome increased dramatically and was reflected in a positive clinical outcome for this patient. Metabolomic analysis revealed that short-chain fatty acids, which have been reported to be associated with improved memory function, were increased after FMT.

Micromonospora pelagivivens sp. nov., a new species of the genus Micromonospora isolated from deep-sea sediment in Japan.

A novel marine actinomycete, designated strain KJ-029T, was isolated from a marine sediment sample (water depth of 226 m) in Kagoshima, Japan. 16S rRNA gene sequence analysis revealed that the new isolate was most closely related to Micromonospora craniellae LHW 63014T (99.3 % similarity). Phylogenetic analyses of the genus Micromonospora based on 16S rRNA gene sequences showed that strain KJ-029T was clustered with Micromonospora craniellae LHW 63014T and Micromonospora endophytica 202201T. However, digital DNA-DNA hybridization analyses presented low levels of relatedness in the range of 24.8-32.9 % between strain KJ-029T and the above closely related strains. The novel strain contained meso-diaminopimelic acid and 3-OH-diaminopimelic acid, d-glutamic acid, glycine and d-alanine in the cell-wall peptidoglycan. The acyl type of the peptidoglycan was glycolyl and mycolic acids were absent. The major menaquinone was MK-9(H4). The whole-cell sugars consisted of glucose, mannose, xylose and ribose. Phosphatidylethanolamine was detected as the major phospholipid and corresponded to phospholipid type II. The predominant cellular fatty acid was iso-C16 : 0. The DNA G+C content of the genomic DNA was 71.5 mol%. Based on the present polyphasic study, strain KJ-029T represents a novel species of the genus Micromonospora, for which the name Micromonospora pelagivivens sp. nov. is proposed. The type strain is KJ-029T (=NBRC 113519T=TBRC 9233T).

Detection of Penicillin G Produced by Penicillium chrysogenum with Raman Microspectroscopy and Multivariate Curve Resolution-Alternating Least-Squares Methods.

Raman microspectroscopy is a minimally invasive technique that can identify molecules without labeling. In this study, we demonstrate the detection of penicillin G inside Penicillium chrysogenum KF425 fungal cells. Raman spectra acquired from the fungal cells had highly overlapped spectroscopic signatures and hence were analyzed with multivariate curve resolution by alternating least-squares (MCR-ALS) to extract the spectra of individual molecular constituents. In addition to detecting spatial distribution of multiple constituents such as proteins and lipids inside the fungal body, we could also observe the subcellular localization of penicillin G. This methodology has the potential to be employed in screening the production of bioactive compounds by microorganisms.

Analysis of a plasmid encoding botulinum neurotoxin type G gene in Clostridium argentinense.

Clostridium argentinense produces botulinum neurotoxin type G (BoNT/G). We sequenced and analyzed the plasmid harboring the bont/G gene, designated pCAG, in C. argentinense strain 2740. The pCAG consisted of 140,070 bp containing the bont/G gene cluster. Although this gene cluster showed high similarities in its DNA sequence and ORF arrangement to those of other bont gene clusters, the other regions of the plasmid did not. A phylogenetic study suggested that pCAG had a unique evolutionary history compared with other clostridial bont-harboring plasmids. This suggests that pCAG is possibly a novel type of plasmid expressing the bont/G gene in C. argentinense.

Saccharopolyspora rhizosphaerae sp. nov., an actinomycete isolated from rhizosphere soil in Thailand.

A novel actinomycete, designated as strain H219T, was isolated from rhizosphere soil collected under an Elephant ear plant (Colocasiaesculenta) in Bangkok, Thailand. Strain H219T was characterised using a polyphasic approach. Phylogenetic analysis of the 16S rRNA gene sequences revealed that this isolate was most closely related to Saccharopolyspora tripterygii JCM 32123T (97.6 %), Saccharopolyspora dendranthemae NBRC 108675T (97.5 %) and Saccharopolyspora flava NBRC 16345T (97.5 %). However, DNA-DNA hybridization analyses showed a low relatedness in the range of 39-48 % between the novel isolate and the above closely related strains. The cell-wall peptidoglycan of strain H219T contained meso-diaminopimelic acid. The diagnostic whole-cell sugars consisted of arabinose and galactose. The cellular fatty acid profile mainly comprised iso-C16 : 0, anteiso-C17 : 0, iso-C15 : 0, and 10-methyl C17 : 0. The major menaquinone was MK-9(H4). The detected phospholipids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylethanolamine-containing hydroxylated fatty acids and an unknown phospholipid. The DNA G+C content was 70.6 mol%. Strain H219T represented chemotaxonomic and morphological characteristics that were consistent with members of the genus Saccharopolyspora. However, strain H219T could be distinguished from closely related strains by several phenotypic properties. Based on the data from the polyphasic studies, we propose that strain H219T is a novel species within the genus Saccharopolyspora, Saccharopolysporarhizosphaerae sp. nov. The type strain is H219T (=TBRC 8564T=NBRC 113388T).

Streptomyces boninensis sp. nov., isolated from soil from a limestone cave in the Ogasawara Islands.

Actinomycete strain K11-0400T was isolated from a soil sample collected in the Ogasawara Islands (also known as the Bonin Islands), Tokyo, Japan. Mature spore chains of strain K11-0400T had more than 20 spores per chain. The strain contained ll-diaminopimelic acid as the diamino acid in whole-cell hydrolysates, and MK-9(H6) and MK-9(H4) were the predominant menaquinones. The polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylinositol, and no diagnostic whole-cell sugar was detected. The G+C content of the genomic DNA was 72 mol%. These morphological and chemical features of strain K11-0400T indicated that it belonged to the genus Streptomyces. Strain K11-0400T showed the highest 16S rRNA gene sequence similarity to Streptomyces naganishii NBRC 12892T (97.58 %). However, the DNA-DNA relatedness value between strain K11-0400T and the related strain was below 70 %. Based on morphological, cultural and physiological characteristics, and DNA-DNA relatedness data, strain K11-0400T should be classified as a new species of the genus Streptomyces, for which the name Streptomyces boninensis sp. nov. is proposed. The type strain of S. boninensis is K11-0400T (=NBRC 113073T, TBRC 7755T).

Actinomadura barringtoniae sp. nov., an endophytic actinomycete isolated from the roots of Barringtonia acutangula (L.) Gaertn.

A novel actinomycete strain, designated GKU 128T, isolated from the roots of an Indian oak tree [Barringtonia acutangula (L.) Gaertn.] at Khao Khitchakut district, Chantaburi province, Thailand, was characterized by using a polyphasic approach. The strain formed a branched substrate and aerial mycelia which differentiated into straight to flexuous chains of smooth-ornamented spores. Analysis of the cell wall revealed the presence of meso-diaminopimelic acid and N-acetylmuramic acid in the peptidoglycan. The whole-cell sugars were glucose, madurose, mannose, rhamnose and ribose. Mycolic acids were absent. The major phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositolmannoside. The predominant menaquinones were MK-9(H6), MK-9(H8), MK-9(H0) and MK-9(H4). The major fatty acids were C16 : 0, C18 : 1ω9c and 10-methyl C18 : 0 (tuberculostearic acid). The genomic DNA G+C content was 70.5 mol%. Based on 16S rRNA gene sequence analysis, strain GKU 128T was closely related to the type strains of Actinomadura nitritigenes NBRC 15918T (99.2 % sequence similarity) and Actinomadura fibrosa JCM 9371T (98.7 %). The levels of DNA-DNA relatedness between strain GKU 128T and the closely related type species were less than 19 %. On the basis of phenotypic and genotypic characteristics, strain GKU 128T could be distinguished from its closely related type strains and represents a novel species of the genus Actinomadura, for which the name Actinomadura barringtoniae sp. nov. (=TBRC 7225T=NBRC 113074T) is proposed.

Saccharomonospora colocasiae sp. nov., an actinomycete isolated from the rhizosphere of Colocasia esculenta.

A non-Streptomyces actinomycete, designated as strain S265T, was isolated from rhizosphere collected under an elephant ear plant (Colocasia esculenta) in Bangkok, Thailand. The taxonomic position of this strain was determined by a polyphasic approach. Strain S265T formed single globose spores on long, branching, aerial hyphae. It produced abundant aerial mycelium with green colour. The cell wall contained meso-diaminopimelic acid, and diagnostic whole-cell sugars were arabinose and galactose. Phosphatidylethanolamine and diphosphatidylglycerol were detected predominantly as polar lipids, whereas mycolic acids were not found. The major menaquinone was MK-9(H4), and principal cellular fatty acids were C15 : 1 B, iso-C16 : 1 H, anteiso-C15 : 0 and C15 : 0 2-OH. The DNA G+C content was 69 mol%. According to phylogenetic analysis, strain S265T was clustered with Saccharomonospora glauca K62T (98.1 %) and Saccharomonosporaviridis DSM 43017T (97.1 %) despite its 16S rRNA gene sequence showing the highest similarity value to that of Saccharomonosporaazurea NA-128T (98.6 %). DNA-DNA relatedness values between strain S265T and the closely related strains were in the range of 7-50 %, thus strengthening the evidence derived from the polyphasic study that strain S265T represents a novel species within the genus Saccharomonospora, for which the name Saccharomonosporacolocasiae sp. nov. is proposed. The type strain is S265T (=TBRC 7235T=NBRC 112945T).

Dictyobacter aurantiacus gen. nov., sp. nov., a member of the family Ktedonobacteraceae, isolated from soil, and emended description of the genus Thermosporothrix.

A mesophilic, Gram-stain-positive, spore-forming bacterium that formed branched mycelia was isolated from paddy soil in Gunung Salak (Mount Salak), West Java, Indonesia. This strain, designated S-27T, grew at temperatures between 20 and 37 °C; the optimum growth temperature was 25 to 30 °C, and no growth was observed at 15 or 45 °C. The pH range for growth was pH 3.5 to 8.6; the optimum pH was 6.0, and no growth was observed at pH 3.0 or 9.2. Strain S-27T was able to hydrolyse polysaccharides such as starch, cellulose and xylan. The G+C content of the DNA of strain S-27T was 55.7 mol%. The major fatty acids were iso-C17 : 0 and C16 : 1 2-OH, and the major menaquinone was MK-9 (H2). The cell wall of strain S-27T contained d-glutamic acid, glycine, l-alanine, d-alanine, l-ornithine and ß-alanine in a molar ratio of 1.0 : 1.6 : 1.4 : 0.6 : 0.9 : 1.1. The polar lipids consisted of phosphatidylglycerol, phosphatidylinositol and two glycolipids. The major cell-wall sugar was arabinose. Detailed phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S-27T belongs to the order Ktedonobacterales and is most closely related to Ktedonobacter racemifer SOSP1-21T (89.6 % sequence identity). On the basis of its chemotaxonomic and phenotypic features and phylogenetic position, we concluded that strain S-27T represents a novel genus and species, for which we propose the name Dictyobacter aurantiacus gen. nov., sp. nov. The type strain of Dictyobacter aurantiacus is strain S-27T (=NBRC 109595T=InaCC B312T). Emendation of the description of the genus Thermosporothrix is also provided.

Proposal of Sphaerimonospora cavernae gen. nov., sp. nov. and transfer of Microbispora mesophila (Zhang et al., 1998) to Sphaerimonospora mesophila comb. nov. and Microbispora thailandensis (Duangmal et al., 2012) to Sphaerimonospora thailandensis comb. nov.

The actinomycete strain N74T, isolated from cave soil, was studied using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain N74T formed a stable, distinct lineage cluster together with Microbispora thailandensis NN276T (99.3% similarity) and Microbispora mesophila JCM 3151T (97.5%). Strain N74T was observed to produce single spherical spores on aerial mycelium as reported for M. mesophila JCM 3151T and M. thailandensis NN276T but different from other known species of the genus Microbispora, which are characterized by pairs of spores on aerial hyphae. Multilocus sequence analyses based on concatenated partial gyrB, rpoB, atpD, recA and 16S rRNA gene sequences showed a clear distinction of strain N74T, M. mesophila JCM 3151T and M. thailandensis NN276T from other members of the genus Microbispora, although the chemotaxonomic characteristics of strain N74T were similar to the genus Microbispora; the cell wall contained meso-diaminopimelic acid and the whole-cell hydrolysate contained madurose as the diagnostic sugar. The major menaquinone was MK-9(H4). The fatty acid profile contained iso-C16:0. On the basis of morphological, chemotaxonomic and phylogenetic evidence, strain N74T is assigned to a novel species of a new genus, for which the name Sphaerimonospora cavernae gen. nov., sp. nov. is proposed. The type strain of Sphaerimonospora cavernae is N74T (=BCC 77604T=NBRC 111481T). It is also proposed that M. mesophila and M. thailandensis be transferred to this genus as Sphaerimonospora mesophila comb. nov. (type strain JCM 3151T=NBRC 14179T=DSM 43048T) and Sphaerimonospora thailandensis comb. nov. (type strain NN276T=BCC 41490T=NBRC 107569T), respectively.

Allokutzneria oryzae sp. nov., isolated from rhizospheric soil of Oryza sativa L.

The taxonomic status of a rhizospheric soil actinomycete, designated R8-39(T), was established using a polyphasic approach. The organism had phenotypic and morphological characteristics consistent with its classification in the genus Allokutzneria. Phylogenetic analysis based on an almost complete 16S rRNA gene sequence showed that the strain formed a monophyletic clade with the type strains of members of the genus Allokutzneria. Strain R8-39(T) displayed the highest levels of 16S rRNA gene sequence similarity to Allokutzneria albata DSM 44149(T) (98.8%) and Allokutzneria multivorans YIM 120521(T) (98.3%). However, the DNA-DNA hybridization values between strain R8-39(T) and A. albata and A. multivorans were clearly below the 70% threshold. The organism was found to have chemical characteristics consistent with its classification in the genus Allokutzneria. Whole-cell hydrolysates contained meso-diaminopimelic acid, arabinose, galactose, glucose, mannose, rhamnose and ribose. The main menaquinone was MK-9(H4). No mycolic acid was detected. The G+C content of the genomic DNA was 71.8 mol%. In addition, strain R8-39(T) had a phenotypic profile that readily distinguished it from recognized representatives of the genus Allokutzneria. It is evident from the combined genotypic and phenotypic properties that strain R8-39(T) represents a novel species of the genus Allokutzneria. The proposed name for this species is Allokutzneria oryzae sp. nov.; the type strain is R8-39(T) ( = BCC 60399(T) = NBRC 109649(T)).