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1.
Two bacterial strains, P0211T and P0213T, were isolated from a sea cucumber culture pond in China. The strains were able to resist high copper levels. These two strains were characterized at the phenotypic, chemotaxonomic, and genomic level. They were completely different colors, but the 16S rRNA genes showed 99.30% similarity. Phylogenetic analysis based on the sequences of the 16S rRNA gene and five housekeeping genes (dnaK, sucC, rpoB, gyrB, and rpoD) supported the inclusion of these strains within the genus Alteromonas, and the two isolated strains formed a group separated from the closest species Alteromonas aestuariivivens KCTC 52655T. Genomic analyses, including average nucleotide identity (ANIb and ANIm), DNA–DNA hybridization (DDH), and the percentage of conserved proteins (POCP), clearly separated strains P0211T and P0213T from the other species within the genus Alteromonas with values below the thresholds for species delineation. The chemotaxonomic features (including fatty acid and polar lipid analysis) of strains P0211T and P0213T also confirmed their differentiation from the related taxa.The results demonstrated that strains P0211T and P0213T represented two novel species in the genus Alteromonas, for which we propose the names Alteromonas flava sp. nov., type strain P0211T (= KCTC 62078T = MCCC 1H00242T), and Alteromonas facilis sp. nov., type strain P0213T (= KCTC 62079T = MCCC 1H00243T).  相似文献   

2.
A pale yellow-colored, moderately halophilic, Gram-negative, catalase- and oxidase-positive, non-sporulating, rod-shaped, motile, aerobic bacterium, designated strain JSM 073008T, was isolated from a sea anemone (Anthopleura xanthogrammica) collected from Naozhou Island, Leizhou Bay, South China Sea. The organism was able to grow with 1–20% (w/v) total salts (optimum, 5–10%), at pH 6.0–10.0 (optimum, pH 7.5) and 10–40°C (optimum, 25–30°C). The major cellular fatty acids were C16:0, C16:1 ω7c/iso-C15:0 2-OH and C18:1 ω7c. The polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol and an unidentified phospholipid. The predominant respiratory quinone was Q-8 and the genomic DNA G + C content was 47.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain JSM 073008T should be assigned to the genus Alteromonas, being most closely related to Alteromonas hispanica F-32T (sequence similarity 96.9%), followed by Alteromonas genovensis LMG 24078T (96.6%) and Alteromonas litorea TF-22T (96.4%). The sequence similarities between the novel isolate and the type strains of other recognized Alteromonas species ranged from 95.9% (with Alteromonas stellipolaris ANT 69aT) to 94.5% (with Alteromonas simiduii BCRC 17572T). The combination of phylogenetic analysis, phenotypic characteristics and chemotaxonomic data supported the view that strain JSM 073008T represents a new species of the genus Alteromonas, for which the name Alteromonas halophila sp. nov. is proposed. The type strain is JSM 073008T (=CCTCC AA 207035T = KCTC 22164T). The authors Yi-Guang Chen and Huai-Dong Xiao have contributed equally to this work.  相似文献   

3.
Exploration of unexplored habitats for novel actinobacteria with high bioactivity potential holds great promise in the search for novel entities. During the course of isolation of actinobacteria from desert soils, four actinobacteria, designated as 5K548T, 7K502T, 16K309T and 16K404T, were isolated from the Karakum Desert and their bioactivity potential as well as taxonomic provenances were revealed by comprehensive genome analyses. Pairwise sequence analyses of the 16S rRNA genes indicated that the four strains are representatives of putatively novel taxa within the prolific actinobacterial genus Saccharopolyspora. The strains have typical chemotaxonomic characteristics of the genus Saccharopolyspora by having meso-diaminopimelic acid as diagnostic diaminoacid, arabinose, galactose and ribose as whole-cell sugars. Consistent with this assignment, all of the isolates contained phosphatidylcholine in their polar lipid profiles and MK-9(H4) as the predominant menaquinone. The sizes of the genomes of the isolates ranged from 6.0 to 10.2 Mb and the associated G + C contents from 69.6 to 69.7 %. Polyphasic characterizations including determination of overall genome relatedness indices revealed that the strains are representatives of four novel species in the genus Saccharopolyspora. Consequently, isolates 5K548T, 7K502T, 16K404T and 16K309T are proposed as novel Saccharopolyspora species for which the names of Saccharopolyspora karakumensis sp. nov., Saccharopolyspora elongata sp. nov., Saccharopolyspora aridisoli sp. nov. and Saccharopolyspora terrae sp. nov. are proposed, respectively. Comprehensive genome analysis for biosynthetic gene clusters showed that the strains have high potential for novel secondary metabolites. Moreover, the strains harbour many antimicrobial resistance genes providing more evidence for their potentiality for bioactive metabolites.  相似文献   

4.
A non-pigmented, motile, Gram-negative bacterium designated H 17T was isolated from a seawater sample collected in Port Phillip Bay (the Tasman Sea, Pacific Ocean). The new organism displayed optimal growth between 4 and 37 °C, was found to be neutrophilic and slightly halophilic, tolerating salt water environments up to 10 % NaCl. Strain H 17T was found to be able to degrade starch and Tween 80 but unable to degrade gelatin or agar. Phosphatidylglycerol (27.7 %) and phosphatidylethanolamine (72.3 %) were found to be the only associated phospholipids. The major fatty acids identified are typical for the genus Alteromonas and include C16:0, C16:1ω7, C17:1ω8 and C18:1ω7. The G+C content of the DNA was found to be 43.4 mol%. A phylogenetic study, based on the 16S rRNA gene sequence analysis and Multilocus Phylogenetic Analysis, clearly indicated that strain H 17T belongs to the genus Alteromonas. The DNA?DNA relatedness between strain H 17T and the validly named Alteromonas species was between 30.7 and 46.4 mol%. Based on these results, a new species, Alteromonas australica, is proposed. The type strain is H 17T (= KMM 6016T = CIP 109921T).  相似文献   

5.
Two moderately halophilic and psychrotolerant new Mycoplasma species were isolated from common cephalopods. Three strains were isolated in pure culture from two individual European flying squid (Todarodes sagittatus), and two individual octopuses (Octopus vulgaris). The strains showed optimal growth at 25 °C and a salinity of 3% (w/v) NaCl. Molecular analyses revealed that the isolates belonged to two new, but phylogenetically related species, divergent from all previously described Mollicutes, representing the first marine isolates of the class, and also the first Mycoplasma strains for which NaCl requirement has been demonstrated. A genome search against all available marine metagenomes and 16S rRNA gene databases indicated that these two species represent a novel non-free-living marine lineage of Mollicutes, specifically associated with marine animals. Morphology and physiology were compatible with other members of this group, and genomic and phenotypic analyses demonstrated that these organisms represent two novel species of the genus Mycoplasma, for which the names Mycoplasma marinum sp. nov. and Mycoplasma todarodis sp. nov. are proposed; the type strains are PET (DSM 105487T, CIP 111404T) and 5HT (DSM 105,488T, CIP 111405T), respectively.  相似文献   

6.
Nine bacterial strains designated MT3-5-12T, MT3-5-27, MTV1-9, S-DT1-15T, S-DT1-34, MTV5-3T, MTV4-17, MTV5-12 and MTV5-13 were isolated from the upper layer (1–5 cm in depth) of tidal flat sediment in Quanzhou Bay, China. The 16S rRNA gene of these strains shared maximum sequence similarities with Aestuariivivens insulae KCTC 42350T of 94.9–97.1%. Phylogenetic analyses based on 16S rRNA gene sequences and 120 conserved concatenated proteins placed these strains in three novel phylogenetic clades affiliated to the genus Aestuariivivens of the family Flavobacteriaceae. Strains MT3-5-12T, MT3-5-27 and MTV1-9 were phylogenetically close to A. insulae KCTC 42350T. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strains MT3-5-12Tand MTV1-9 and A. insulae KCTC 42350T were estimated to be 78.5-78.7% and 22.5%, respectively. Strains S-DT1-15T and S-DT1-34 formed a distinctly separated clade from A. insulae KCTC 42350T. The ANI and dDDH values between strains S-DT1-15T and S-DT1-34 and A. insulae KCTC 42350T were 76.3–76.4% and 20.4–20.5%, respectively. The other four strains MTV5-3T, MTV4-17, MTV5-12 and MTV5-13, formed a third novel clade, distinctly separated from A. insulae KCTC 42350T. The ANI and dDDH values between strains MTV5-3T and MTV4-17 and A. insulae KCTC 42350T were 74.7% and 19.1–19.2%, respectively. The phylogenetic analyses and whole genomic comparisons, combined with phenotypic and chemotaxonomic features, strongly supported the nine strains could be classified as three novel species within the genus Aestuariivivens, for which the names Aestuariivivens marinum sp. nov. MT3-5-12T, Aestuariivivens sediminis sp. nov. S-DT1-15T, and Aestuariivivens sediminicola sp. nov. MTV5-3T are proposed.  相似文献   

7.
Spectra of five isolates (LMG 28358T, LMG 29879T, LMG 29880T, LMG 28359T and R-53705) obtained from gut samples of wild bumblebees of Bombus pascuorum, Bombus lapidarius and Bombus terrestris were grouped into four MALDI-TOF MS clusters. RAPD analysis revealed an identical DNA fingerprint for LMG 28359T and R-53705 which also grouped in the same MALDI-TOF MS cluster, while different DNA fingerprints were obtained for the other isolates.Comparative 16S rRNA gene sequence analysis of the four different strains identified Gilliamella apicola NCIMB 14804T as nearest neighbour species. Average nucleotide identity values of draft genome sequences of the four isolates and of G. apicola NCIMB 14804T were below the 96% threshold value for species delineation and all four strains and G. apicola NCIMB 14804T were phenotypically distinct. Together, the draft genome sequences and phylogenetic and phenotypic data indicate that the four strains represent four novel Gilliamella species for which we propose the names Gilliamella intestini sp. nov., with LMG 28358T as the type strain, Gilliamella bombicola sp. nov., with LMG 28359T as the type strain, Gilliamella bombi sp. nov., with LMG 29879T as the type strain and Gilliamella mensalis sp. nov., with LMG 29880T as the type strain.  相似文献   

8.
A Gram-negative, non-motile, non-endospore-forming bacterial strain, designated DPSR-4T, was isolated from a tidal flat sediment on the southern coast of Korea. Strain DPSR-4T grew optimally at 25–30°C, at pH 7.0–7.5 and in the presence of 2% (w/v) NaCl. A Neighbour-Joining phylogenetic tree based on 16S rRNA gene sequences revealed that strain DPSR-4T clustered with Salinimonas chungwhensis BH030046T by a high bootstrap resampling value of 99.7%. Strain DPSR-4T exhibited 96.2% 16S rRNA gene sequence similarity to that of S. chungwhensis BH030046T and 93.7–96.6% sequence similarity to the sequences of type strains of Alteromonas species. Strain DPSR-4T contained Q-8 as the predominant ubiquinone and iso-C15:0 2-OH and/or C16:1 ω7c, C16:0 and C18:1 ω7c as the major fatty acids. The major polar lipids detected in strain DPSR-4T and S. chungwhensis KCTC 12239T were phosphatidylglycerol, phosphatidylethanolamine and an unidentified phospholipid. The DNA G+C content was 53.4 mol%. Differential phenotypic properties and phylogenetic distinctiveness of strain DPSR-4T demonstrated that this strain is distinguishable from the sole recognized species of the genus Salinimonas, S. chungwhensis. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain DPSR-4T is considered to represent a novel species of the genus Salinimonas, for which the name Salinimonas lutimaris sp. nov. is proposed. The type strain is DPSR-4T (KCTC 23464T, CCUG 60743T).  相似文献   

9.
Three strains, H01100409BT, H01100413B, and H27100402HT, were isolated from several internal organs of diseased redbanded seabream (Pagrus auriga) reared in Andalusia (Southern Spain). All strains were studied by phenotypic, including chemotaxonomy, and genomic characteristics. Phylogenetic analysis based on concatenated sequences of six housekeeping genes (gyrB, ftsZ, topA, mreB, gapA, and 16S rRNA) supported the inclusion of the strains within the clade Phosphoreum of the genus Photobacterium, and two of the strains (H27100402HT and H01100409BT) formed a tight group separated from the closest species P. aquimaris. Genomic analyses, including average nucleotide identity (ANIb and ANIm) and DNA–DNA hybridization (DDH), clearly separated strains H27100402HT and H01100409BT from the other species within the clade Phosphoreum with values below the thresholds for species delineation. The chemotaxonomic features (including FAME analysis and MALDI-TOF-MS) of H27100402HT and H01100409BT strains confirmed their differentiation from the related taxa. The results demonstrated that strain H01100413B was classified as P. aquimaris and the strains H27100402HT and H01100409BT represented a new species each in the genus Photobacterium, for which we propose the names Photobacterium malacitanum sp. nov., type strain H27100402HT (=CECT 9190T = LMG 29992T), and Photobacterium andalusiense sp. nov., type strain H01100409BT (=CECT 9192T = LMG 29994T).  相似文献   

10.
Two filamentous actinomycetes isolated from a hay meadow soil were provisionally assigned to the genus Streptomyces based on morphological features. The isolates were found to have chemical and morphological properties typical of the genus Streptomyces and formed distinct phyletic lines in the 16S rRNA gene tree. Isolate I36T was most closely related to Streptomyces glauciniger NBRC 100913T and isolate I37T to Streptomyces mirabilis NBRC 13450T. Low DNA:DNA relatedness values were recorded between each of the isolates and their closest phylogenetic neighbour. The isolates were also distinguished from their nearest phylogenetic neighbour, and from one another, using a combination of phenotypic properties. These data indicate that the isolates should be recognised as new species in the genus Streptomyces. The names proposed for these new taxa are Streptomyces erringtonii sp. nov. and Streptomyces kaempferi sp. nov. with isolate I36T (=CGMCC 4.7016T = KACC 15424T) and isolate I37T (=CGMCC 4.7020T = KACC 15428T) as the respective type strains.  相似文献   

11.
Bacteria in the family Geobacteraceae have been proven to fill important niches in a diversity of anaerobic environments and global biogeochemical processes. Here, three bacterial strains in this family, designated Red875T, Red259T, and Red421T were isolated from river sediment and paddy soils in Japan. All of them are Gram-staining-negative, strictly anaerobic, motile, flagellum-harboring cells that form red colonies on agar plates and are capable of utilizing Fe(III)-NTA, Fe(III) citrate, ferrihydrite, MnO2, fumarate, and nitrate as electron acceptors with acetate, propionate, pyruvate, and glucose as electron donors. Phylogenetic analysis based on the 16S rRNA gene and 92 concatenated core proteins sequences revealed that strains Red259T and Red421T clustered with the type strains of Geomonas species, whereas strain Red875T formed an independent lineage within the family Geobacteraceae. Genome comparison based on  average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values clearly distinguished these three strains from other Geobacteraceae members, with lower values than the thresholds for species delineation. Moreover, strain Red875T also shared low average amino acid identity (AAI) and percentage of conserved proteins (POCP) values with the type species of the family Geobacteraceae. Based on these physiological, chemotaxonomic, and phylogenetic distinctions, we propose that strain Red875T (=NBRC 114290T = MCCC 1K04407T) represents a novel genus in the family Geobacteraceae, namely, Geomesophilobacter sediminis gen. nov., sp. nov., and strains Red259T (=NBRC 114288T = MCCC 1K05016T) and Red421T (=NBRC 114289T = MCCC 1K06216T) represent two novel independent species in the genus Geomonas, namely, Geomonas propionica sp. nov. and Geomonas anaerohicana sp. nov., respectively.  相似文献   

12.
13.
Two bacterial strains, KIS66-7T and 5GH26-15T, were isolated from soil samples collected in the South Korean cities of Tongyong and Gongju, respectively. Both strains were aerobic, Gram-stain-positive, mesophilic, flagellated, and rodshaped. A phylogenetic analysis revealed that both strains belonged to the family Microbacteriaceae of the phylum Actinobacteria. The 16S rRNA gene sequence of strain KIS66-7T had the highest similarities with those of Labedella gwakjiensis KSW2-17T (97.3%), Cryobacterium psychrophilum DSM 4854T (97.2%), Leifsonia lichenia 2SbT (97.2%), Leifsonia naganoensis JCM 10592T (97.0%), and Cryobacterium mesophilum MSL-15T (97.0%). Strain 5GH26-15T showed the highest sequence similarities with Leifsonia psychrotolerans LI1T (97.4%) and Schumannella luteola KHIAT (97.1%). The 16S rRNA gene sequence from KIS66-7T exhibited 96.4% similarity with that from 5GH26-15T. Strain KIS66-7T contained a B2γ type peptidoglycan structure with D-DAB as the diamino acid; MK-13, MK-12, and MK-14 as the respiratory quinones; ai-C15:0, ai-C17:0, and i-C16:0 as the major cellular fatty acids; and diphosphatidylglycerol, phatidylglycerol, and glycolipids as the predominant polar lipids. Strain 5GH26-15T had a B2β type peptidoglycan structure with D-DAB as the diamino acid; MK-14 and MK-13 as the respiratory quinones; ai-C15:0, i-C16:0, and ai-C{vn17:0} as the major cellular fatty acids; and diphosphatidylglycerol, phatidylglycerol, and glycolipids as the predominant polar lipids. Both strains had low DNA-DNA hybridization values (<40%) with closely related taxa. Based on our polyphasic taxonomic characterization, we propose that strains KIS66-7T and 5GH26-15T represent novel genera and species, for which we propose the names Diaminobutyricibacter tongyongensis gen. nov., sp. nov. (type strain KIS66-7T=KACC 15515T=NBRC 108724T) and Homoserinibacter gongjuensis gen. nov., sp. nov. (type strain 5GH26-15T=KACC 15524T=NBRC 108755T) within the family Microbacteriaceae.  相似文献   

14.
Current -omics methods allow the collection of a large amount of information that helps in describing the microbial diversity in nature. Here, and as a result of a culturomic approach that rendered the collection of thousands of isolates from 5 different hypersaline sites (in Spain, USA and New Zealand), we obtained 21 strains that represent two new Salinibacter species. For these species we propose the names Salinibacter pepae sp. nov. and Salinibacter grassmerensis sp. nov. (showing average nucleotide identity (ANI) values < 95.09% and 87.08% with Sal. ruber M31T, respectively). Metabolomics revealed species-specific discriminative profiles. Sal. ruber strains were distinguished by a higher percentage of polyunsaturated fatty acids and specific N-functionalized fatty acids; and Sal. altiplanensis was distinguished by an increased number of glycosylated molecules. Based on sequence characteristics and inferred phenotype of metagenome-assembled genomes (MAGs), we describe two new members of the genus Salinibacter. These species dominated in different sites and always coexisted with Sal. ruber and Sal. pepae. Based on the MAGs from three Argentinian lakes in the Pampa region of Argentina and the MAG of the Romanian lake Fără Fund, we describe the species Salinibacter pampae sp. nov. and Salinibacter abyssi sp. nov. respectively (showing ANI values 90.94% and 91.48% with Sal. ruber M31T, respectively). Sal. grassmerensis sp. nov. name was formed according to the rules of the International Code for Nomenclature of Prokaryotes (ICNP), and Sal. pepae, Sal. pampae sp. nov. and Sal. abyssi sp. nov. are proposed following the rules of the newly published Code of Nomenclature of Prokaryotes Described from Sequence Data (SeqCode). This work constitutes an example on how classification under ICNP and SeqCode can coexist, and how the official naming a cultivated organism for which the deposit in public repositories is difficult finds an intermediate solution.  相似文献   

15.
The taxonomic positions of five Gram-negative, non-spore-forming and non-motile bacterial strains isolated from the rhizosphere of sand dune plants were examined using a polyphasic approach. The analysis of the 16S rRNA gene sequence indicated that all of the isolates fell into four distinct phylogenetic clusters belonging to the genus Chryseobacterium of the family Flavobacteriaceae. The 16S rRNA gene sequence similarities of isolates to mostly related type strains of Chryseobacterium ranged from 97.5% to 98.5%. All strains contained MK-6 as the predominant menaquinone, and iso-C15:0, iso-C17:0 3-OH and a summed feature of iso-C15:0 2-OH and/or C16:1 ω7c as the dominant fatty acids. Combined phenotypic, genotypic and chemotaxonomic data supported that they represented four novel species in the genus Chryseobacterium, for which the names Chryseobacterium hagamense sp. nov. (type strain RHA2-9T=KCTC 22545T=NBRC 105253T), Chryseobacterium elymi sp. nov. (type strain RHA3-1T=KCTC 22547T=NBRC 105251T), Chryseobacterium lathyri sp. nov. (type strain RBA2-6T=KCTC 22544T=NBRC 105250T), and Chryseobacterium rhizosphaerae sp. nov. (type strain RSB3-1T=KCTC 22548T=NBRC 105248T) are proposed.  相似文献   

16.
17.
Bartonella is one of the noncore bacterial genera in the honey bee (Apis mellifera) gut. So far, only one species, Bartonella apis, has been described from the honey bee gut. Previous analyses based on the genomic information of isolates and metagenome-assembled genomes suggested the existence of multiple Bartonella species in the bee guts. Here, 10 strains were isolated and characterized from the gut of A. mellifera from Jilin Province, China. New isolates shared >95% 16S rRNA gene sequence similarity with other species of the genus Bartonella. Phylogenetic analysis revealed that new isolates clustered with other type strains of Bartonella, and the bee gut Bartonella could be classified into three clades. The in silico DDH and average nucleotide identity values between strains of different clusters from the honey bee gut are 29.1–32.5% and 87.6–89.3%, all below the recommended 70.0% and 95% cutoff points. Cells are Gram-staining-negative rods and can grow on the surface of Brain Heart Infusion agar plates supplemented with defibrinated sheep blood in an aerobic environment with 5% CO2 at 35–37 °C. Strains from different species varied in both phenotypic and chemotaxonomic characterizations. Comparative genomic analysis indicated that B. choladocola had unique sets of genes encoding invasin, representing the potential for this species to both live as a gut symbiont and also as an erythrocytic pathogen. Thus, we propose two novel species Bartonella choladocola sp. nov. whose type strain is W8125T(=JCM 35030T = ACCC 62057T), and Bartonella apihabitans sp. nov. whose type strain is W8097T(=JCM 35029T = ACCC 62056T).  相似文献   

18.
The phenotypic and genotypic characteristics of fourteen human clinical Achromobacter strains representing four genogroups which were delineated by sequence analysis of nusA, eno, rpoB, gltB, lepA, nuoL and nrdA loci, demonstrated that they represent four novel Achromobacter species. The present study also characterized and provided two additional reference strains for Achromobacter ruhlandii and Achromobacter marplatensis, species for which, thus far, only single strains are publicly available, and further validated the use of 2.1% concatenated nusA, eno, rpoB, gltB, lepA, nuoL and nrdA sequence divergence as a threshold value for species delineation in this genus. Finally, although most Achromobacter species can be distinguished by biochemical characteristics, the present study also highlighted considerable phenotypic intraspecies variability and demonstrated that the type strains may be phenotypically poor representatives of the species. We propose to classify the fourteen human clinical strains as Achromobacter mucicolens sp. nov. (with strain LMG 26685T [=CCUG 61961T] as the type strain), Achromobacter animicus sp. nov. (with strain LMG 26690T [=CCUG 61966T] as the type strain), Achromobacter spiritinus sp. nov. (with strain LMG 26692T [=CCUG 61968T] as the type strain), and Achromobacter pulmonis sp. nov. (with strain LMG 26696T [=CCUG 61972T] as the type strain).  相似文献   

19.
Twelve Mycoplasma (M.) strains isolated from the nose, the trachea, and the lung of ostriches (Struthio camelus) displaying respiratory disease were investigated. Analysis of 16S rRNA gene sequences placed five of these strains within the M. synoviae cluster, and seven strains within the M. hominis cluster of genus Mycoplasma, which was further confirmed by analyses of the 16S-23S rRNA intergenic spacer region, and partial rpoB gene and amino acid sequences. Genomic information as well as phenotypic features obtained by matrix-assisted laser desorption ionization time of flight (MALDI-ToF) mass spectrometry analysis and serological reactions indicated that the strains examined are representatives of two hitherto unclassified species of genus Mycoplasma, for which the names Mycoplasma nasistruthionis sp. nov., with type strain 2F1AT (= ATCC BAA-1893T = DSM 22456T), and Mycoplasma struthionis sp. nov., with type strain 237IAT (= ATCC BAA-1890T = DSM 22453T), are proposed.  相似文献   

20.
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