scholarly journals From Metagenomics to Pure Culture: Isolation and Characterization of the Moderately Halophilic Bacterium Spiribacter salinus gen. nov., sp. nov.

2014 ◽  
Vol 80 (13) ◽  
pp. 3850-3857 ◽  
Author(s):  
María José León ◽  
Ana B. Fernández ◽  
Rohit Ghai ◽  
Cristina Sánchez-Porro ◽  
Francisco Rodriguez-Valera ◽  
...  

ABSTRACTRecent metagenomic studies on saltern ponds with intermediate salinities have determined that their microbial communities are dominated by bothEuryarchaeotaand halophilic bacteria, with a gammaproteobacterium closely related to the generaAlkalilimnicolaandArhodomonasbeing one of the most predominant microorganisms, making up to 15% of the total prokaryotic population. Here we used several strategies and culture media in order to isolate this organism in pure culture. We report the isolation and taxonomic characterization of this new, never before cultured microorganism, designated M19-40T, isolated from a saltern located in Isla Cristina, Spain, using a medium with a mixture of 15% salts, yeast extract, and pyruvic acid as the carbon source. Morphologically small curved cells (young cultures) with a tendency to form long spiral cells in older cultures were observed in pure cultures. The organism is a Gram-negative, nonmotile bacterium that is strictly aerobic, non-endospore forming, heterotrophic, and moderately halophilic, and it is able to grow at 10 to 25% (wt/vol) NaCl, with optimal growth occurring at 15% (wt/vol) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence comparison showed that strain M19-40Thas a low similarity with other previously described bacteria and shows the closest phylogenetic similarity with species of the generaAlkalilimnicola(94.9 to 94.5%),Alkalispirillum(94.3%), andArhodomonas(93.9%) within the familyEctothiorhodospiraceae. The phenotypic, genotypic, and chemotaxonomic features of this new bacterium showed that it constitutes a new genus and species, for which the nameSpiribacter salinusgen. nov., sp. nov., is proposed, with strain M19-40T(= CECT 8282T= IBRC-M 10768T= LMG 27464T) being the type strain.

2013 ◽  
Vol 63 (Pt_3) ◽  
pp. 844-848 ◽  
Author(s):  
Maryam Bagheri ◽  
Mohammad Ali Amoozegar ◽  
Peter Schumann ◽  
Maryam Didari ◽  
Malihe Mehrshad ◽  
...  

A novel Gram-stain-positive, moderately halophilic bacterium, designated strain G8BT, was isolated from water of the hypersaline lake Aran-Bidgol in Iran and characterized taxonomically using a polyphasic approach. Cells of strain G8BT were rod-shaped, motile and produced oval endospores at a terminal position in swollen sporangia. Strain G8BT was strictly aerobic, catalase-positive and oxidase-negative. The strain was able to grow at NaCl concentrations of 0.5–12.5 % (w/v), with optimum growth occurring at 5–7.5 % (w/v) NaCl. The optimum temperature and pH for growth were 35–40 °C and pH 7.5–8.0, respectively. On the basis of 16S rRNA gene sequence analysis, strain G8BT was shown to belong to the genus Ornithinibacillus within the phylum Firmicutes and showed closest phylogenetic similarity with Ornithinibacillus bavariensis WSBC 24001T (97.6 %). The DNA G+C content of strain G8BT was 36.9 mol%. The major cellular fatty acids of strain G8BT were anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0 and iso-C16 : 0, and its polar lipid pattern consisted of phosphatidylglycerol, diphosphatidylglycerol, four unknown phospholipids and an unknown aminolipid. The isoprenoid quinones were MK-7 (98 %) and MK-8 (2 %). Strain G8BT contained a peptidoglycan of type A4β, l-Orn–d-Asp. All these features confirmed the placement of isolate G8BT within the genus Ornithinibacillus . DNA–DNA hybridization experiments revealed a low level of relatedness (6 %) between strain G8BT and Ornithinibacillus bavariensis DSM 15681T. On the basis of evidence from this study, a novel species of the genus Ornithinibacillus , Ornithinibacillus halophilus sp. nov., is proposed, with strain G8BT ( = IBRC-M 10683T = KCTC 13822T) as the type strain.


2015 ◽  
Vol 65 (Pt_4) ◽  
pp. 1354-1359 ◽  
Author(s):  
Carmen Infante-Domínguez ◽  
Cristina Sánchez-Porro ◽  
Antonio Ventosa

A yellow-pigmented, motile, Gram-stain-negative, moderately halophilic and strictly aerobic bacterium, designated BA42AL-1T, was isolated from water of a saltern of Santa Pola, Alicante, Spain. Strain BA42AL-1T grew in media containing 5–20 % (w/v) salts (optimum 7.5 % salts). It grew between pH 6.0 and 9.0 (optimally at pH 7.5) and at 15–45 °C (optimally at 37 °C). Phylogenetic analysis based on the comparison of 16S rRNA gene sequences revealed that strain BA42AL-1T is a member of the genus Aquisalimonas . The closest relatives to this strain were Aquisalimonas halophila YIM 95345T and Aquisalimonas asiatica CG12T with sequence similarities of 99.4 % and 97.0 %, respectively. DNA–DNA hybridization between the novel isolate and Aquisalimonas halophila YIM 95345T revealed a relatedness of 54 %. The major fatty acids of strain BA42AL-1T were C18 : 1ω6c/C18 : 1ω7c, C19 : 0 cyclo ω8c and C16 : 0, and lower contents of C12 : 0 and C18 : 0. The polar lipid pattern of strain BA42AL-1T consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmethylethanolamine, phosphatidylcholine, two glycolipids, a lipid and four unknown phospholipids. The G+C content of the genomic DNA of this strain was 65.0 mol%. Based on the DNA–DNA hybridization, phenotypic, chemotaxonomic and phylogenetic data presented in this study, strain BA42AL-1T is proposed as a novel species of the genus Aquisalimonas , for which the name Aquisalimonas lutea sp. nov. is suggested. The type strain is BA42AL-1T ( = CCM 8472T = CECT 8326T = LMG 27614T).


2014 ◽  
Vol 64 (Pt_7) ◽  
pp. 2210-2216 ◽  
Author(s):  
Yun-Jiao Zhang ◽  
Man Jia ◽  
Yong-Chun Ma ◽  
Kai-Yang Lu ◽  
Fei Tian ◽  
...  

A Gram-negative, moderately halophilic, strictly aerobic strain, designated YIM 95345T, was isolated from a soil sample of a hypersaline mine in Yunnan province, PR China, and subjected to a polyphasic taxonomic study. Strain YIM 95345T grew at 15–45 °C (optimum 30–35 °C), 3.0–23.0 % (w/v) NaCl (optimum 10.0–11.0 %, w/v) and pH 6.0–9.0 (optimum pH 7.0–8.0). Phylogenetic analyses based on 16S rRNA gene sequences revealed that the organism belongs to the genus Aquisalimonas and exhibited sequence similarity of 96.6 % to the sole type strain Aquisalimonas asiatica CG12T. The predominant isoprenoid quinone was Q-8 and the major fatty acids were C16 : 0, C19 : 0 cyclo ω8c and C18 : 1ω7c. The polar lipids consisted of diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, three aminolipids and three unidentified phospholipids. The G+C content of the genomic DNA was 59.4 mol%. Based on the results of our comparative phylogenetic, chemotaxonomic and physiological analyses, the new isolate is assigned to a novel species of the genus Aquisalimonas , for which the name Aquisalimonas halophila sp. nov. is proposed, with the type strain YIM 95345T ( = DSM 25902T = CCTCC AB 2012043T).


2012 ◽  
Vol 62 (Pt_8) ◽  
pp. 1857-1863 ◽  
Author(s):  
Pankyung Kim ◽  
Jae-Chan Lee ◽  
Dong-Jin Park ◽  
Kee-Sun Shin ◽  
Jeong-Yoon Kim ◽  
...  

A Gram-staining-positive, moderately halophilic bacterium, designated strain BH097T, was isolated from solar saltern soil of Bigeum Island in south-west Korea. Cells were motile rods, producing spherical endospores at a terminal position in swollen sporangia. Strain BH097T was strictly aerobic, grew at pH 5.5–9.5 (optimum, pH 8.0), at 10–52 °C (optimum, 37 °C) and at salinities of 1–22 % (w/v) NaCl (optimum, 7 % NaCl). On the basis of 16S rRNA gene sequence analysis, strain BH097T was shown to belong to the genus Gracilibacillus within the phylum Firmicutes , and showed closest sequence similarity to Gracilibacillus saliphilus DSM 19802T (95.8 %), Gracilibacillus thailandensis TP2-8T (95.6 %), Gracilibacillus boraciitolerans DSM 17256T (95.5 %), ‘Gracilibacillus quinghaiensis’ DSM 17858 (95.4 %) and Gracilibacillus halophilus DSM 17856T (95.2 %). The DNA G+C content of this novel isolate was 37.9 mol%. The major cellular fatty acids of strain BH097T were anteiso-C15 : 0, iso-C15 : 0 and C16 : 0, and its polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol two unknown phospholipids and a glycolipid. The isoprenoid quinone was MK-7, and the peptidoglycan type was A1γ, with meso-diaminopimelic acid as the diagnostic diamino acid. On the basis of polyphasic evidence from this study, strain BH097T represents a novel species of the genus Gracilibacillus for which the name Gracilibacillus bigeumensis sp. nov. is proposed. The type strain is BH097T ( = KCTC 13130T = DSM 19028T).


2020 ◽  
Vol 70 (4) ◽  
pp. 2254-2260 ◽  
Author(s):  
Shehzad Abid Khan ◽  
Sepideh Zununi Vahed ◽  
Haleh Forouhandeh ◽  
Vahideh Tarhriz ◽  
Nader Chaparzadeh ◽  
...  

In the course of screening halophilic bacteria in Urmia Lake in Iran, which is being threatened by dryness, a novel Gram-negative, moderately halophilic, heterotrophic and short rod-shaped bacteria was isolated and characterized. The bacterium was isolated from a water specimen and designated as TBZ3T. Colonies were found to be creamy yellow, with catalase- and oxidase-positive activities. The growth of strain TBZ3T was observed to be at 10–45 °C (optimum, 30 °C), at pH 6.0–9.0 (optimum, pH 7.0) and in the presence of 0.5–20 % (w/v) NaCl (optimum, 7.5 %). Strain TBZ3T contained C16 : 0, cyclo-C19 : 0  ω8c, summed feature 3 (comprising C16 : 1  ω7c and/or C16 : 1  ω6c) and summed feature 8 (comprising C18 : 1  ω7c and/or C18 : 1  ω6c) as major fatty acids and ubiquinone-9 as the only respiratory isoprenoid quinone. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, glycolipid, unidentified phospholipid and unidentified polar lipids were detected as the major polar lipids. Strain TBZ3T was found to be most closely related to Halomonas saccharevitans AJ275T , Halomonas denitrificans M29T and Halomonas sediminicola CPS11T with the 16S rRNA gene sequence similarities of 98.93, 98.15 and 97.60 % respectively and in phylogenetic analysis strain TBZ3T grouped with Halomonas saccharevitans AJ275T contained within a large cluster within the genus Halomonas . Based on phenotypic, chemotaxonomic and molecular properties, strain TBZ3T represents a novel species of the Halomonas genus, for which the name Halomonas urmiana sp. nov. is proposed. The type strain is TBZ3T (=DSM 22871T=LMG 25416T).


2012 ◽  
Vol 62 (Pt_9) ◽  
pp. 2174-2179 ◽  
Author(s):  
Yun-Jiao Zhang ◽  
Shu-Kun Tang ◽  
Rong Shi ◽  
Hans-Peter Klenk ◽  
Chao Chen ◽  
...  

A Gram-stain-negative, moderately halophilic, strictly aerobic bacterium, designated YIM 95161T, was isolated from brine of a salt well in Yunnan province, China, and subjected to a taxonomic study using a polyphasic approach. Cells of strain YIM 95161T were short rods, approximately 0.9–1.4 µm long and 0.4–0.6 µm wide. Strain YIM 95161T grew at 15–40 °C (optimum, 25–30 °C), 6–29 % (w/v) NaCl (optimum, 14–19 %) and at pH 5.0–8.0 (optimum, pH 7.0). The predominant isoprenoid quinone was Q-8. The major fatty acids (>10 %) were summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c) and C14 : 0. The polar lipids consisted of diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, one unknown phosphoglycolipid and two unknown lipids. The DNA G+C content was 69.5 %. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 95161T was a member of the genus Salinisphaera and exhibited sequence similarities of 96.7 %, 95.6 % and 95.4 % to Salinisphaera shabanensis E1L3AT, Salinisphaera dokdonensis CL-ES53T and Salinisphaera hydrothermalis EPR70T, respectively. On the basis of phylogenetic, physiological and chemotaxonomic analysis, strain YIM 95161T represents a novel species of the genus Salinisphaera , for which the name Salinisphaera halophila sp. nov. is proposed. The type strain is YIM 95161T ( = CCTCC AB 2011132T = JCM 17431T).


Author(s):  
Auttaporn Booncharoen ◽  
Wonnop Visessanguan ◽  
Nattakorn Kuncharoen ◽  
Supalurk Yiamsombut ◽  
Pannita Santiyanont ◽  
...  

An aerobic, Gram-stain-positive, endospore-forming, rod-shaped and moderately halophilic strain SKP4-6T, was isolated from shrimp paste (Ka-pi) collected from Samut Sakhon Province, Thailand. Phylogenetic analysis revealed that strain SKP4-6T belonged to the genus Halobacillus and was most closely related to Halobacillus salinus JCM 11546T (98.6 %), Halobacillus locisalis KCTC 3788T (98.6 %) and Halobacillus yeomjeoni KCTC 3957T (98.6 %) based on 16S rRNA gene sequence similarity. The digital DNA–DNA hybridization (dDDH) and average nucleotide identity (ANI) values between strain SKP4-6T and its related species were 18.2–19.3 % and 69.84–84.51 %, respectively, which were lower than the threshold recommended for species delineation. The strain grew optimally at 30–40 °C, at pH 7.0 and with 10–15 % (w/v) NaCl. It contained l-Orn–d-Asp in the cell wall peptidoglycan. The DNA G+C content was 44.8 mol%. The major fatty acids were iso-C15 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The predominant isoprenoid quinone was MK-7. Phosphatidylglycerol and diphosphatidylglycerol were present as major polar lipids. Based on this polyphasic approach, digital DNA–DNA relatedness and ANI values, strain SKP4-6T represents a novel species of the genus Halobacillus , for which the name Halobacillus fulvus sp. nov. is proposed. The type strain is SKP4-6T (=JCM 32624T=TISTR 2595T).


2013 ◽  
Vol 63 (Pt_10) ◽  
pp. 3710-3714 ◽  
Author(s):  
Ying-Yi Huo ◽  
Lin Xu ◽  
Chun-Sheng Wang ◽  
Jun-Yi Yang ◽  
Hong You ◽  
...  

A Gram-stain-negative, aerobic, moderately halophilic bacterium, strain DY53T, was isolated from a deep-seawater sample collected from the eastern Pacific Ocean. This isolate grew in the presence of 0.5–10.0 % (w/v) NaCl, at pH 6.5–8.5 and at 15–40 °C. The optimum NaCl concentration for growth of DY53T was 2 % (w/v) at 35 °C. Chemotaxonomic analysis showed MK-7 as the predominant menaquinone and iso-C15 : 0, summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1ω7c), iso-C15 : 1 G, iso-C15 : 0 3-OH and iso-C17 : 0 3-OH as major cellular fatty acids. The genomic DNA G+C content was 40.8 mol%. Phylogenetic trees based on 16S rRNA gene sequences revealed that Fabibacter halotolerans UST030701-097T was the closest neighbour, with 96.7 % sequence similarity. Based on phylogenetic, chemotaxonomic and phenotypic data, we propose that strain DY53T represents a novel species of the genus Fabibacter , for which the name Fabibacter pacificus sp. nov. is proposed. The type strain is DY53T( = CGMCC 1.12402T = JCM 18885T).


2015 ◽  
Vol 65 (Pt_12) ◽  
pp. 4434-4440 ◽  
Author(s):  
Su-Jin Kim ◽  
Jae-Chan Lee ◽  
Song-Ih Han ◽  
Kyung-Sook Whang

A Gram-staining-positive, moderately halophilic bacterium, designated strain NGS-2T, was isolated from sediment of a solar saltern pond located in Shinan, Korea. Strain NGS-2T was a strictly aerobic, non-motile rod that grew at pH 5.0–10.0 (optimum, pH 8.0), at 10–30 °C (optimum, 28 °C) and in the presence of 1–20 % (w/v) NaCl (optimum, 10 % NaCl). Phylogenetic analyses based on 16S rRNA gene sequences indicated that strain NGS-2T belonged to the genus Halobacillus, with sequence similarity of 98.4–95.8 % to existing type strains, showing the highest sequence similarity to Halobacillus dabanensis D-8T (98.4 %), H. litoralis SL-4T (98.4 %), H. trueperi SL-5T (98.2 %), H. faecis IGA7-4T (98.2 %), H. profundi IS-Hb4T (98.1 %) and H. mangrovi MS10T (98.0 %). The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, phosphatidyl-N-methylethanolamine and an unknown glycolipid. The cell-wall peptidoglycan was based on l-Orn–d-Asp, the predominant isoprenoid quinone was menaquinone 7 (MK-7) and the major fatty acids were anteiso-C15: 0 and anteiso-C17: 0. The DNA G+C content of the novel isolate was 45.0 mol%. Levels of DNA–DNA relatedness between strain NGS-2T and the type strains of 12 other species of the genus ranged from 32 to 3 %. On the basis of the polyphasic analysis conducted in this study, strain NGS-2T represents a novel species of the genus Halobacillus, for which the name Halobacillus sediminis sp. nov. is proposed. The type strain is NGS-2T ( = KACC 18263T = NBRC 110639T).


Author(s):  
Kuppusamy Pandiyan ◽  
Prity Kushwaha ◽  
Samadhan Y. Bagul ◽  
Hillol Chakdar ◽  
Munusamy Madhaiyan ◽  
...  

A moderately halophilic, Gram-stain-negative, aerobic bacterium, strain D1-1T, belonging to the genus Halomonas , was isolated from soil sampled at Pentha beach, Odisha, India. Phylogenetic trees reconstructed based on 16S rRNA genes and multilocus sequence analysis of gyrB and rpoD genes revealed that strain D1-1T belonged to the genus Halomonas and was most closely related to Halomonas alimentaria YKJ-16T (98.1 %) followed by Halomonas ventosae Al12T (97.5 %), Halomonas sediminicola CPS11T (97.5 %), Halomonas fontilapidosi 5CRT (97.4 %) and Halomonas halodenitrificans DSM 735T (97.2 %) on the basis of 16S rRNA gene sequence similarity. Sequence identities with other species within the genus were lower than 97.0 %. The digital DNA–DNA hybridization (dDDH) and average nucleotide identity (ANI) values of 22.4–30 % and 79.5–85.4 % with close relatives of H. halodenitrificans DSM 735T, H . alimentaria YKJ-16T, H. ventosae Al12T and H. fontilapidosi 5CRT were lower than the threshold recommended for species delineation (70 % and 95–96 % for dDDH and ANI, respectively). Further, strain D1-1T formed yellow-coloured colonies; cells were rod-shaped, motile with optimum growth at 30 °C (range, 4–45 °C) and 2–8 % NaCl (w/v; grew up to 24 % NaCl). The major fatty acids were summed feature 8 (C18 : 1  ω7c/C18 : 1  ω6c), summed feature 3 (C16 : 1  ω7c/C16 : 1  ω6c) and C16 : 0 and the main respiratory quinone was ubiquinone Q-9 in line with description of the genus. Based on its chemotaxonomic and phylogenetic characteristics and genome uniqueness, strain D1-1T represents a novel species in the genus Halomonas , for which we propose the name Halomonas icarae sp. nov., within the family Halomonadaceae . The type strain is D1-1T (=JCM 33602T=KACC 21317T=NAIMCC-B-2254T).


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