scholarly journals Nocardiopsis aegyptia sp. nov., isolated from marine sediment

2004 ◽  
Vol 54 (2) ◽  
pp. 453-456 ◽  
Author(s):  
Soraya A. Sabry ◽  
Nevine B. Ghanem ◽  
Gehan A. Abu-Ella ◽  
Peter Schumann ◽  
Erko Stackebrandt ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Marine Sediment ◽  
Sequence Similarity ◽  
Aerial Mycelium ◽  
Diaminopimelic Acid ◽  
Rrna Gene ◽  
16S Rdna Sequence Analysis ◽  
Light Yellow ◽  
Dna Reassociation ◽  
Abu Qir Bay

An actinomycete, strain SNG49T, was isolated from marine sediment of Abu Qir Bay, on the western seashore of Alexandria, Egypt. The bacterium was aerobic and Gram-positive. It produced beige to light-yellow aerial mycelium, brown substrate mycelium and straight to flexuous hyphae, but no specific spore chains. 16S rDNA sequence analysis and chemotaxonomic markers were consistent with classification of strain SNG49T in the genus Nocardiopsis, i.e. meso-diaminopimelic acid; no diagnostic sugars; phosphatidylcholine, phosphatidylmethylethanolamine, phosphatidylinositol, phosphatidylglycerol and diphosphatidylglycerol as polar lipids; menaquinones of the MK-10 series from MK-10(H0) to MK-10(H8); and iso/anteiso-branched and 10-methyl-branched fatty acids, the principal fatty acids being anteiso-17 : 0 and tuberculostearic acid. Nocardiopsis lucentensis and Nocardiopsis alba are the phylogenetic neighbours of strain SNG49T, respectively showing 98·8 and 98·7 % 16S rRNA gene sequence similarity; however, moderate DNA–DNA reassociation values between these two species and strain SNG49T (44 and 60 %, respectively) showed that strain SNG49T could be clearly separated from them. These data, together with distinct physiological traits, led to the conclusion that this isolate represents a novel species within the genus Nocardiopsis, for which the name Nocardiopsis aegyptia is proposed. The type strain is SNG49T (=DSM 44442T=NRRL B-24244T).

scholarly journals Actinokineospora guangxiensis sp. nov., isolated from soil

2015 ◽  
Vol 65 (Pt_12) ◽  
pp. 4650-4654 ◽  
Author(s):  
Hao Wu ◽  
Bin Liu
Keyword(s):  
Fatty Acids ◽  
Sequence Similarity ◽  
Aerial Mycelium ◽  
Diaminopimelic Acid ◽  
Rrna Gene ◽  
Gene Sequence Analysis ◽  
Hydroxylated Fatty Acids ◽  
Pr China ◽  
Diamino Acid ◽  
Unknown Phospholipid

A novel actinomycete, designated strain GK-6T, was isolated from a soil sample from Nanning, Guangxi province, PR China. The strain grew at 20–40 °C, pH 6.0–11.0 and with 0–7.0 % NaCl. It formed well-developed aerial and vegetative mycelia. The aerial mycelium was white and the vegetative mycelium was yellow. The long branching aerial mycelia yielded rod-shaped arthrospores, the spores had smooth surfaces and were non-motile. Strain Gk-6T contained meso-diaminopimelic acid as the diagnostic diamino acid, the whole-cell sugars were galactose, glucose and arabinose. Major fatty acids were iso-C16 : 0, iso-C15 : 0 and C17 : 0. MK-9(H4) was the predominant menaquinone. The polar phospholipids were phosphatidylethanolamine, phosphatidylethanolamine-containing hydroxylated fatty acids, diphosphatidylglycerol, ninhydrin-positive glycophospholipid and an unknown phospholipid. The G+C content of the genomic DNA was 73.4 mol%. The 16S rRNA gene sequence analysis indicated that the organism was a member of the genus Actinokineospora and its closest relative among recognized species was Actinokineospora soli JCM 17695T (97.7 % sequence similarity). But the phenotypic characteristics of strain Gk-6T were significantly different from those of A. soli JCM 17695T, and DNA–DNA hybridization showed low relatedness (22.6–28.3 %) between strain Gk-6T and JCM 17695T. On the basis of the phenotypic and phylogenetic data, strain Gk-6T represents a novel species of the genus Actinokineospora, and the name Actinokineospora guangxiensis sp. nov. is proposed. The type strain is Gk-6T ( = DSM 46779 T = CGMCC 4.7154T).

scholarly journals Thermoactinomyces guangxiensis sp. nov., a thermophilic actinomycete isolated from mushroom compost

2015 ◽  
Vol 65 (Pt_9) ◽  
pp. 2859-2864 ◽  
Author(s):  
Hao Wu ◽  
Bin Liu ◽  
Shangli Pan
Keyword(s):  
Fatty Acids ◽  
Sequence Similarity ◽  
Aerial Mycelium ◽  
Diaminopimelic Acid ◽  
Rrna Gene ◽  
Mushroom Compost ◽  
Dna Relatedness ◽  
Thermophilic Actinomycete ◽  
Thermoactinomyces Vulgaris ◽  
Laceyella Putida

A novel thermophilic actinomycete, designated strain CD-1T, was isolated from mushroom compost in Nanning, Guangxi province, China. The strain grew at 37–55 °C (optimum 45–50 °C), pH 6.0–11.0 (optimum pH 7.0–9.0) and with 0–2.0  % NaCl (optimum 0–1.0  %), formed well-developed white aerial mycelium and pale-yellow vegetative mycelium, and single endospores (0.8–1.0 μm diameter) were borne on long sporophores (2–3 μm length). The endospores were spherical-polyhedron in shape with smooth surface. Based on its phenotypic and phylogenetic characteristics, strain CD-1T is affiliated to the genus Thermoactinomyces. It contained meso-diaminopimelic acid as the diagnostic diamino acid; the whole-cell sugars were ribose and glucose. Major fatty acids were iso-C15  :  0, C16  :  0, anteiso-C15  :  0 and iso-C17  :  0. MK-7 was the predominant menaquinone. The polar phospholipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylethanolamine containing hydroxylated fatty acids, ninhydrin-positive glycophospholipid, an unknown phospholipid and glycolipids. The G+C content of the genomic DNA was 48.8  %. 16S rRNA gene sequence analysis showed that the organism was closely related to Lihuaxuella thermophila YIM 77831T (95.69  % sequence similarity), Thermoactinomyces daqus H-18T (95.49  %), Laceyella putida KCTC 3666T (95.05  %), Thermoactinomyces vulgaris KCTC 9076T (95.01  %) and Thermoactinomyces intermedius JCM 3312T (94.55  %). Levels of DNA–DNA relatedness between strain CD-1T and Lihuaxuella thermophila JCM 18059T, Thermoactinomyces daqus DSM 45914T, Laceyella putida JCM 8091T, Thermoactinomyces vulgaris JCM 3162T and Thermoactinomyces intermedius JCM 3312T were low (22.8, 33.3, 24.7, 29.4 and 30.0  %, respectively). A battery of phenotypic, genotypic and DNA–DNA relatedness data indicated that strain CD-1T represented a novel species of the genus Thermoactinomyces, for which the name Thermoactinomyces guangxiensis sp. nov. is proposed. The type strain is CD-1T ( = ATCC BAA-2630T = CGMCC 4.7156T).

scholarly journals Nocardiopsis dassonvillei subsp. crassaminis subsp. nov., isolated from freshwater sediment, and reappraisal of Nocardiopsis alborubida Grund and Kroppenstedt 1990 emend. Nouioui et al. 2018

2020 ◽  
Vol 70 (12) ◽  
pp. 6172-6179
Author(s):  
Miladis Isabel Camacho Pozo ◽  
Anneleen D. Wieme ◽  
Suyén Rodríguez Pérez ◽  
Gabriel Llauradó Maury ◽  
Cindy Snauwaert ◽  
...  
Keyword(s):  
Type Species ◽  
Sequence Similarity ◽  
Aerial Mycelium ◽  
Diaminopimelic Acid ◽  
Freshwater Sediment ◽  
Rrna Gene ◽  
Content Type ◽  
Link Type ◽  
Light Yellow ◽  
Actinomycete Strain

An actinomycete, strain D1T, was isolated from a freshwater sediment sample collected from the San Pablo river in the La Risueña community, Santiago de Cuba province, Cuba. The strain was identified as a member of the genus Nocardiopsis by means of a polyphasic taxonomic study. It produced a light yellow non-fragmented substrate mycelium, a white well-developed aerial mycelium and straight to flexuous hyphae. No specific spore chains were observed. Strain D1T contained meso-diaminopimelic acid, no diagnostic sugars, and MK-10(H2), MK-10(H4), MK-10 and MK-10(H6) as predominant menaquinones, but not phosphatidylcholine as diagnostic polar lipid of the genus Nocardiopsis . The predominant fatty acids were iso-C16 : 0, 10-methyl-C18 : 0 and anteiso-C17 : 0. Strain D1T showed the highest degree of 16S rRNA gene sequence similarity to Nocardiopsis synnematoformans DSM 44143T (99.8 %), Nocardiopsis dassonvillei subsp. albirubida NBRC 13392T (99.8 %) and Nocardiopsis dassonvillei subsp. dassonvillei DSM 43111T (99.6 %). A genomic OrthoANIu value between D1T and N. dassonvillei subsp. dassonvillei DSM 43111T of 97.63 % and a dDDH value of 78.9 % indicated that strain D1T should be classified in N. dassonvillei . However, phenotypic characteristics distinguished strain D1T from its nearest neighbour taxon. On basis of these results we propose to classify strain D1T (=LMG 30468T=CECT 30033T) as a representative of a novel subspecies of the genus Nocardiopsis , for which the name Nocardiopsis dassonvillei subsp. crassaminis subsp. nov. is proposed. In addition, the genomic distance between N. dassonvillei subsp. albirubida NBRC 13392T and N. dassonvillei subsp. dassonvillei DSM 43111T as determined through OrthoANIu (93.64 %) and dDDH (53.40 %), along with considerable phenotypic and chemotaxonomic differences reported in earlier studies, indicated that the classification of this taxon as Nocardiopsis alborubida Grund and Kroppenstedt 1990 is to be preferred over its classification as N. dassonvillei subsp. albirubida Evtushenko et al. 2000.

scholarly journals Streptomyces pharmamarensis sp. nov. isolated from a marine sediment

2012 ◽  
Vol 62 (Pt_5) ◽  
pp. 1165-1170 ◽  
Author(s):  
Lorena Carro ◽  
Paz Zúñiga ◽  
Fernando de la Calle ◽  
Martha E. Trujillo
Keyword(s):  
Fatty Acids ◽  
Marine Sediment ◽  
Type Species ◽  
Gene Sequence ◽  
Sequence Similarity ◽  
Diaminopimelic Acid ◽  
Physiological Data ◽  
Rrna Gene ◽  
Content Type ◽  
Link Type

A Gram-stain-positive actinobacterium, strain PM267T, was isolated from a marine sediment sample in the Mediterranean Sea. The novel strain produced extensively branched substrate and aerial hyphae that carried spiral spore chains. Substrate and aerial mycelia were cream–white and white, respectively. Diffusible pigments were not observed. 16S rRNA gene sequence analysis revealed that strain PM267T belonged to the genus Streptomyces and shared a gene sequence similarity of 97.1 % with Streptomyces artemisiae YIM 63135T and Streptomyces armeniacus JCM 3070T. Values <97 % were obtained with other sequences representing members of the genus Streptomyces . The cell wall peptidoglycan contained ll -diaminopimelic acid. MK-9(H8) was the major menaquinone. The phospholipid pattern included phosphatidylethanolamine as diagnostic lipid (type II). Major fatty acids found were iso- and anteiso- fatty acids. The G+C content of the DNA was 71.2 mol%. The strain was halotolerant and was able to grow in the presence of 9 % (w/v) NaCl (with an optimum of 2 %). On the basis of these results and additional physiological data obtained in the present study, strain PM267T represents a novel species within the genus Streptomyces for which the name Streptomyces pharmamarensis sp. nov. is proposed (type strain PM267T  = CECT 7841T  = DSM 42032T).

Longimycelium tulufanense gen. nov., sp. nov., a filamentous actinomycete of the family Pseudonocardiaceae

2013 ◽  
Vol 63 (Pt_8) ◽  
pp. 2813-2818 ◽  
Author(s):  
Zhan-Feng Xia ◽  
Tong-Wei Guan ◽  
Ji-Sheng Ruan ◽  
Ying Huang ◽  
Li-Li Zhang
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Sequence Similarity ◽  
Aerial Mycelium ◽  
Diaminopimelic Acid ◽  
Rrna Gene ◽  
Content Type ◽  
North West ◽  
Link Type ◽  
The Family

A novel filamentous actinomycete strain, designated TRM 46004T, was isolated from sediment of Aiding Lake in Tulufan Basin (42° 64′ N 89° 26′ E), north-west China. The isolate was characterized using a polyphasic approach. The isolate formed abundant aerial mycelium with few branches and vegetative mycelium, occasionally twisted and coiled; spherical sporangia containing one to several spherical spores developed at the ends of short sporangiophores on aerial mycelium. The G+C content of the DNA was 65.2 mol%. The isolate contained meso-diaminopimelic acid as the diagnostic diamino acid and xylose, galactose and ribose as the major whole-cell sugars. The diagnostic phospholipids were phosphatidylethanolamine, phosphatidylcholine and phosphatidylglycerol. The predominant menaquinones were MK-9(H4), MK-9(H6) and MK-9(H10). The major fatty acids were iso-C16 : 0 and anteiso-C17 : 0. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain TRM 46004T formed a distinct lineage within the family Pseudonocardiaceae and showed 91.7–96.1 % 16S rRNA gene sequence similarity with members of the family Pseudonocardiaceae . On the basis of the evidence from this polyphasic study, a novel genus and species, Longimycelium tulufanense gen. nov., sp. nov., are proposed. The type strain of Longimycelium tulufanense is TRM 46004T ( = CGMCC 4.5737T = NBRC 107726T).

scholarly journals Streptomyces fenghuangensis sp. nov., isolated from seawater

2011 ◽  
Vol 61 (12) ◽  
pp. 2811-2815 ◽  
Author(s):  
Honghui Zhu ◽  
Shumei Jiang ◽  
Qing Yao ◽  
Yonghong Wang ◽  
Meibiao Chen ◽  
...  
Keyword(s):  
Gene Sequence ◽  
Sequence Similarity ◽  
Aerial Mycelium ◽  
Diaminopimelic Acid ◽  
Rrna Gene ◽  
Substrate Mycelium ◽  
Physiological And Biochemical Characteristics ◽  
Type Strains ◽  
Physiological And Biochemical ◽  
The 16S Rrna Gene

An actinomycete, designated strain GIMN4.003T, was isolated from seawater collected in Sanya, China. It produced white aerial mycelium and yellow substrate mycelium on Gause’s synthetic agar medium no. 1. The substrate mycelium colour was not sensitive to pH. Scanning electron microscopy observations revealed that GIMN4.003T produced straight to flexuous spore chains of rough to warty spores. ll-Diaminopimelic acid was present in the cell-wall hydrolysate. Based on chemotaxonomy and morphological features, strain GIMN4.003T was identified as a member of the genus Streptomyces. Melanin was not produced. No antimicrobial activity was detected against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Penicillium citrinum or Candida albicans. Analysis of the 16S rRNA gene sequence revealed that the highest sequence similarity was to Streptomyces radiopugnans R97T (99.0 %). However, DNA relatedness between GIMN4.003T and S. radiopugnans DSM 41901T was low (41.24±1.47 %). Furthermore, the morphological, physiological and biochemical characteristics of strain GIMN4.003T were different from those of S. radiopugnans DSM 41901T and the type strains of other closely related Streptomyces species. On the basis of its physiological and molecular properties, it is evident that strain GIMN4.003T ( = CCTCCM 208215T  = NRRL B-24801T) represents the type strain of a novel species within the genus Streptomyces, for which the name Streptomyces fenghuangensis sp. nov. is proposed.

scholarly journals Nocardiopsis arvandica sp. nov., isolated from sandy soil

2011 ◽  
Vol 61 (5) ◽  
pp. 1189-1194 ◽  
Author(s):  
Javad Hamedi ◽  
Fatemeh Mohammadipanah ◽  
Gabriele Pötter ◽  
Cathrin Spröer ◽  
Peter Schumann ◽  
...  
Keyword(s):  
Sandy Soil ◽  
Novel Species ◽  
Sequence Similarity ◽  
Phylogenetic Analyses ◽  
Aerial Mycelium ◽  
Diaminopimelic Acid ◽  
Rrna Gene ◽  
Hybridization Data ◽  
Actinomycete Strain ◽  
Taxonomic Approach

An actinomycete strain, HM7T, was isolated from sandy soil from the banks of the Arvand River, Khoramshahr, Iran. In a study conducted using a polyphasic taxonomic approach, it was found that this strain represented a novel species in the genus Nocardiopsis. Strain HM7T had long, branched substrate mycelia and the aerial mycelium completely fragmented to long chains of rod-shaped spores. The cell wall of strain HM7T contained meso-diaminopimelic acid, galactose and ribose. The predominant phospholipids were phosphatidylglycerol, phosphatidylethanolamine diphosphatidylglycerol, and phosphatidylcholine. The main menaquinones were MK-10(H2), MK-10(H4), MK-10(H0) and MK-9(H2). Strain HM7T showed the highest degree of 16S rRNA gene sequence similarity with Nocardiopsis sinuspersici UTMC 00102T (99.8 %) and Nocardiopsis quinghaiensis YIM 28A4T (99.3 %), but differed significantly from these type strains based on DNA–DNA hybridization data (37.6 and 38.9 % relatedness, respectively). Physiological, biochemical and phenotypic characteristics allowed strain HM7T to be differentiated from related species. On the basis of results from phenotypic and phylogenetic analyses, strain HM7T is considered to represent a novel species of the genus Nocardiopsis, for which the name Nocardiopsis arvandica sp. nov. is proposed; the type strain of this species is HM7T ( = UTMC 00103T = DSM 45278T = CCUG 58831T).

scholarly journals Nocardiopsis nikkonensis sp. nov., isolated from a compost sample

2010 ◽  
Vol 60 (12) ◽  
pp. 2967-2971 ◽  
Author(s):  
Hideki Yamamura ◽  
Shin-ya Ohkubo ◽  
Yuumi Ishida ◽  
Misa Otoguro ◽  
Tomohiko Tamura ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Sequence Similarity ◽  
Aerial Mycelium ◽  
Morphological Characterization ◽  
Diaminopimelic Acid ◽  
Rrna Gene ◽  
Compost Sample ◽  
Actinomycete Strain ◽  
Biochemical Analyses

An actinomycete strain, designated YU1183-22T, was isolated from a compost sample collected in Nikko, Japan. The isolate formed white aerial mycelium with relatively long aerial hyphae showing chains of arthrospores. Strain YU1183-22T grew with 0–10 % (w/v) NaCl, at pH 6–11 and at 10–37 °C (optimum 30 °C). Strain YU1183-22T contained meso-diaminopimelic acid and no diagnostic sugars. The predominant menaquinones were MK-10(H10) and MK-10(H8). The polar lipids were phosphatidylcholine and phosphatidylglycerol. The major cellular fatty acids were iso-C16 : 0, anteiso-C17 : 0 and tuberculostearic acid. The G+C content of the DNA was 72.3 mol%. Chemotaxonomic and morphological characterization clearly demonstrated that strain YU1183-22T belonged to the genus Nocardiopsis. Phylogenetic analysis using 16S rRNA gene sequences showed that the isolate was closely related to Nocardiopsis salina YIM 90010T (98.0 % 16S rRNA gene sequence similarity), Nocardiopsis xinjiangensis YIM 90004T (97.9 %) and Nocardiopsis kunsanensis HA-9T (97.3 %). However, DNA–DNA relatedness as well as physiological and biochemical analyses showed that strain YU1183-22T could be differentiated from its closest phylogenetic relatives. It is proposed that this strain be classified as a representative of a novel species of the genus Nocardiopsis, with the name Nocardiopsis nikkonensis sp. nov. The type strain is YU1183-22T (=NBRC 102170T =KCTC 19666T).

scholarly journals Streptomyces lopnurensis sp. nov., an actinomycete isolated from soil

2014 ◽  
Vol 64 (Pt_12) ◽  
pp. 4179-4183 ◽  
Author(s):  
Bei Zheng ◽  
Xiao-Xue Han ◽  
Zhan-Feng Xia ◽  
Chuan-Xing Wan ◽  
Li-Li Zhang
Keyword(s):  
Sequence Similarity ◽  
Aerial Mycelium ◽  
Diaminopimelic Acid ◽  
Rrna Gene ◽  
Content Type ◽  
Link Type ◽  
Gram Staining ◽  
Lop Nur ◽  
Nacl Concentration ◽  
Diamino Acid

A novel actinomycete, designated strain TRM 49590T, was isolated from a soil sample from Lop Nur in Xinjiang Province, China. Strain TRM 49590T was aerobic, Gram-staining-positive, with an optimum NaCl concentration for growth of 1.5 % (w/v) and an optimum temperature for growth of 28–37 °C. The aerial mycelium was sparse, cylindrical and smooth-surfaced with irregular branches on ISP medium 4. The whole-cell sugars of strain TRM 49590T were ribose and glucose. The diagnostic diamino acid contained ll-diaminopimelic acid. The predominant menaquinones were MK-9(H6) and MK-9(H8), with MK-9(H4) and MK-10(H6) present in smaller amounts. The major fatty acids were iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The G+C content of the genomic DNA was 62.2 mol%. The major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and phosphatidylinositol mannoside. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain TRM 49590T belongs to the genus Streptomyces with a sequence similarity of 97.16 % with the most closely related species Streptomyces sodiiphilus . Based on these observations, strain TRM 49590T is proposed to represent a novel species of the genus Streptomyces for which the name Streptomyces lopnurensis sp. nov. is suggested. The type strain is TRM 49590T ( = CCTCC AA 2013018T = NRRL B59109T).

Nocardioides szechwanensis sp. nov. and Nocardioides psychrotolerans sp. nov., isolated from a glacier

2013 ◽  
Vol 63 (Pt_1) ◽  
pp. 129-133 ◽  
Author(s):  
Qing Liu ◽  
Yu-hua Xin ◽  
Hong-can Liu ◽  
Yu-guang Zhou ◽  
Ying Wen
Keyword(s):  
Fatty Acids ◽  
Type Strain ◽  
Sequence Similarity ◽  
Diaminopimelic Acid ◽  
Rrna Gene ◽  
Content Type ◽  
Cellular Fatty Acids ◽  
Link Type ◽  
Pr China ◽  
Spore Forming Bacteria

Two Gram-positive, rod-shaped, non-spore-forming bacteria (strains RHLT1-17T and RHLT2-1T) were isolated from Hailuogou glacier in Szechwan province, PR China. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strains belonged to the genus Nocardioides and shared 97.8 % sequence similarity with each other and 97.6 and 98.4 % sequence similarity, respectively, with Nocardioides kribbensis KSL-2T. Strain RHLT1-17T grew at 0–35 °C and strain RHLT2-1T grew at 0–25 °C. The major cellular fatty acids of strain RHLT1-17T were C17 : 1ω8c (32.69 %) and iso-C16 : 0 (21.74 %). The major cellular fatty acids of strain RHLT2-1T were C18 : 1ω9c (28.72 %), summed feature 3 (17.14 %; comprising C16 : 1ω7c and/or C16 : 1ω6c), iso-C16 : 0 (14.35 %), C16 : 0 (9.96 %) and iso-C14 : 0 (8.34 %). Both strains contained ll-2,6-diaminopimelic acid in the cell-wall peptidoglycan and MK-8(H4) as the predominant menaquinone. On the basis of data obtained using a polyphasic approach, two novel species, Nocardioides szechwanensis sp. nov. (type strain RHLT1-17T  = CGMCC 1.11147T  = NBRC 108562T) and Nocardioides psychrotolerans sp. nov. (type strain RHLT2-1T  = CGMCC 1.11156T  = NBRC 108563T), are proposed.

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