Three strictly anaerobic, methanogenic strains JH1 T , JH4 and JH8 were isolated from rumen of the Korean native cattle (HanWoo; Bos taurus coreanae) in South Korea. The colonies were circular, opaque, and slightly yellowish. Phylogenetic analyses of 16S rRNA gene and mcrA (encoding a subunit of methyl-coenzyme M reductase) sequences confirmed the affiliation of the novel strains with the Methanobacteriales, and Methanobrevibacter wolinii SH T was the most closely related species. The 16S rRNA gene and mcrA sequence similarities between strains JH1 T , JH4 and JH8 and M. wolinii SH T were 96.2 and 89.0 % respectively, and DNA-DNA hybridization of the isolates and M. wolinii DSM 11976 T showed a 20 % reassociation. Strain JH1 T exhibited 92 % DNA-DNA relatedness with strains JH4 and JH8, and their 16S rRNA gene and mcrA sequences were identical. Cells stained Gram-positive and were non-motile rods, 1.5-1.8 mm long and 0.6 mm wide. The strains were able to use H 2 /CO 2 and formate. The optimum temperature and pH ranges for growth were 37-40 6C and pH 6.5-7.0. The DNA G+C content of strain JH1 T was 28 mol%. Based on data from this study using a polyphasic approach, the three strains represent a novel species of genus Methanobrevibacter, for which the name Methanobrevibacter boviskoreani sp. nov. is proposed. The type strain is JH1 T (5KCTC 4102 T 5JCM 18376 T ).
A strictly anaerobic, Gram-negative, non-spore-forming bacterium, designated GH1T , was isolated from the rumen of Korean native cattle (HanWoo). Cells were straight to slightly curved rods (2.0-4.5 mm long) and were motile by peritrichous flagella. The isolate grew at 30-45 6C (optimum 40 6C), at pH 5.5-6.5 (optimum pH 6.0) and with up to 3.5 % (w/v) NaCl. Strain GH1T produced acid from D-glucose, D-ribose and D-xylose, with butyric acid being the major end product. The genomic DNA G+C content was 54.6 mol%. Based on comparative 16S rRNA gene sequence analysis, strain GH1 T was most closely related to Oscillibacter valericigenes Diverse and competitive micro-organisms inhabit the rumen and belong to three domains: Bacteria, Archaea (methanogens) and Eucarya (protozoa and fungi) (Miron et al., 2001;Pers-Kamczyc et al., 2011). This complex community plays a very important role in the degradation of cellulosic feeds and the supply of nutrients to the host in the form of volatile fatty acids (Kamra, 2005;Van Soest, 1994). The structure and ecology of the whole microbial community in rumen should be more completely understood in order to manipulate rumen fermentation or control the level of bio-emission of methane from ruminants. A number of micro-organisms in rumen are still non-culturable but their individual function could be critical to the ruminal ecosystem (Pers-Kamczyc et al., 2011). Therefore, isolation and characterization of novel rumen micro-organisms should be performed for complete understanding of ruminal ecology.Korean native cattle, 'HanWoo' (Bos taurus coreanae), inhabit in the region of Korea peninsula and produce good-quality meat. Understanding their physiology, nutrition, metabolism and reproduction is of prime importance for the Korean native cattle industry as it is a significant contributor to farming households, the national economy and the security of food stuffs in Korea (Hwang et al., 2000).In our study, we tried to isolate diverse bacteria from the rumen of HanWoo using strictly anaerobic cultivation technique. Fresh rumen fluid (900 ml) was collected 4 h after morning feeding from a ruminally cannulated 1-yearold HanWoo steer fed 4 kg concentrate and 0.5 kg rice straw (dry matter) twice daily at a research farm of Seoul Abbreviation: DMA, dimethylacetal.
Two novel bacterial strains, GH2-4 T and GH2-5, were isolated from mangrove soil near the seashore of Weno island in Chuuk state, Micronesia, and were characterized by a polyphasic approach. The two strains were strictly aerobic, Gram-staining-positive, motile, endospore-forming rods that were catalase-and oxidase-positive. Colonies were circular, convex, stringy and transparent yellowish (GH2-4 T ) or opaque whitish (GH2-5). The 16S rRNA gene sequences of the two isolates were identical. The most closely related strains in terms of 16S rRNA gene sequence similarity were Bacillus kochii WCC 4582 T , B. horneckiae DSM 23495 T , B. azotoformans LMG 9581 T , B. cohnii DSM 6307 T and B. halmapalus DSM 8723 T (95.6, 95.4, 95.4, 95.2 and 95.2 % similarity, respectively). The partial groEL sequence of strain GH2-4 T was identical to that of strain GH2-5 and showed ,85 % similarity to those of the most closely related strains. The isolates grew at pH 5-12 (optimal growth at pH 9), at 10-40 6C (optimum 30-35 6C) and at 0-9 % (w/v) NaCl (optimum 1-3 % NaCl). The cell-wall peptidoglycan of strains GH2-4 T and GH2-5 contained meso-diaminopimelic acid and cell-wall hydrolysates contained ribose as a major sugar. The DNA G+C content was 36 mol%, and DNA-DNA relatedness between the isolates and five related reference strains was 20-24 %. Strain GH2-4 T exhibited 81 % DNA-DNA relatedness with strain GH2-5. The major cellular fatty acids of both strains were iso-C 15 : 0 , iso-C 16 : 0 , iso-C 14 : 0 and anteiso-C 15 : 0 and the predominant menaquinone was MK-7. On the basis of the evidence from this polyphasic study, strains GH2-4 T and GH2-5 (5KCTC 331435JCM 189955DSM 27084) represent a novel species of the genus Bacillus, for which the name Bacillus solimangrovi sp. nov. is proposed; the type strain is GH2-4 T (5KCTC 33142 T 5JCM 18994 T 5DSM 27083 T ).Mangroves are woody plants located at the interface between land and sea. They represent a typical ecosystem in tropical and subtropical coastal biomes, and they play significant roles as feeding, breeding and refuge areas for diverse organisms and maintain a considerable food web based on their detritus (Holguin et al., 2001;Kathiresan & Bingham, 2001). The mangrove ecosystem is distinguished from others because of periodic tidal flooding and changeable environmental factors, such as salinity and nutrient availability (Alongi, 1988;Holguin et al., 2006). There have been many reports regarding the isolation of novel micro-organisms from mangrove-related environments. At the time of writing, 63 genera and 113 species had been newly described as novel micro-organisms isolated from mangrove environments, based on searches of PubMed (http://www.ncbi.nlm.nih. gov/). They include species of the yeast genus Candida The genus Bacillus, in the phylum Firmicutes, is a large group of rod-shaped, endospore-forming bacteria that includes more than 200 species (Euzéby, 2010). Since the genus Bacillus was described by Cohn in 1872, the number of species in the genus has increased considerabl...
A novel facultatively anaerobic strain DH1(T) was isolated from deep sub-seafloor sediment at a depth of 900 m below the seafloor off Seo-do (the west part of Dokdo Island) in the East Sea of the Republic of Korea. The new strain was characterized using polyphasic approaches. The isolate was Gram-stain-negative, motile by gliding, non-spore-forming rods, oxidase-negative, and catalase-positive; and formed colonies of orange-red color. The NaCl range for growth was 0.5-7.0% (w/v) and no growth was observed in the absence of NaCl. The isolate grew optimally at 30°C, with 2% (w/v) NaCl and at pH 7. The cell-wall hydrolysates contained ribose as a major sugar. The DNA G+C content was 40.8 mol%. The closest related strains are Sunxiuqinia faeciviva JAM-BA0302(T) and Sunxiuqinia elliptica DQHS-4(T) (97.9 and 96.3% sequence similarity, respectively). The level of DNA-DNA relatedness between strain DH1(T) and S. faeciviva JAM-BA0302(T) was around 41% (but only 6% between DH1T and S. elliptica DQHS-4(T)). The major cellular fatty acids of the isolate were contained iso-C15:0 (25.9%), anteiso-C15:0 (16.7%), and summed feature 9 (comprising C16:0 3-OH and/or unknown fatty acid of dimethylacetal ECL 17.157; 13.2%). The predominant menaquinone was MK-7. On the basis of polyphasic evidence from this study, the isolate was considered to represent a novel species of the genus Sunxiuqinia, for which the name Sunxiuqinia dokdonensis sp. nov. is proposed; the type strain is DH1(T) (=KCTC 32503(T) =CGMCC 1.12676(T) =JCM 19380(T)).
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