Isolation and Characterization of Halophilic and Halotolerant Bacteria from Urmia Lake after the Recent Drought Disaster in 2015

2020 ◽  
Vol 9 (2) ◽  
pp. 111-119
Author(s):  
Elham Kazemi ◽  
Vahideh Tarhriz ◽  
Mohammad S. Hejazi ◽  
Mohammad A. Amoozegar

Background: Due to the last two decades of drought disaster, which resulted in the loss of the main part of Urmia Lake water and changed the natural conditions of an environment, especially ionic strength. Objective: We aimed to isolate and characterize halophilic and halotolerant bacteria in Urmia Lake, Iran, 2015. Urmia Lake is a permanent and salty inland lake located in the Azerbaijan region in northwestern Iran. Methods: Sampling was carried out in multiple water-filled locations of the lake. Liquid basal media for the enrichment of bacteria was successively applied and colonies were isolated by the plating method. Isolates were then distinguished based on differences in colony, Gram staining, microscopic shape, and biochemical properties. Results: One chemolithotrophic isolate belonging to Thiobacillus thioparus and 41 heterotrophic isolates were obtained. The 16S rRNA gene sequences showed that all 42 isolates belong to the genera Kocuria (21.42%), Marinobacter (11.90%), Micrococcus (11.90%), Thalassobacillus (11.90%), Bacillus (11.90%), Halomonas (7.14%) and Thiobacillus (2.38%). Conclusion: Based on 16S rRNA similarity, 5 of 41 isolates showed the potential to be introduced as new species. The dominant genera with abounded frequency were found to be Kocuria, Bacillus and Thalassobacillus genera.

Author(s):  
Priscilla Mercy Anitha D ◽  
Periyar Selvam Sellamuthu ◽  
Seema A Kulkarni

Objective: The aim of this study was to isolate, characterize and identify the potential lactic acid bacteria from fermented ragi millet (Eleusine coracana).Methods: A total of 177 isolates were isolated from the fermented ragi porridge. All the isolates were subjected to preliminary screening (Gram staining and catalase test). The physiological features of the probiotic isolates such as tolerance to pH (2.0, 4.0), phenol (0.3%, 0.5%) and bile salt (0.3%) were carried out. Antibiotic susceptibility test was performed using disc diffusion method. Free radical scavenging assay was conducted by 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. The isolates were further characterized for their cholesterol-lowering property. Based on their survival ability and biochemical tests, the selected potent isolates were identified by 16S rRNA gene sequence by PCR and phylogenetic analysis. The 16S rRNA gene sequencing confirmed that isolates were Lactococcus lactis RS01, Lactobacillus plantarum RS09, Lactobacillus plantarum RS16 and Lactobacillus plantarum RS23.Results: Twenty five isolates were screened from the 177 total isolates based on their morphological and biochemical characteristics (Gram staining and catalase test). All the isolates showed tolerance against acid, phenol and bile salts. However, RS09 and RS23 exhibited maximum viable count and demonstrated good tolerance at the end of 3 h. Highest antioxidant activity was recorded in RS09 ranging from 32-85%. Among the tested strains, the degradation rate of supernatants, RS09 (61.9%), RS23 (60%) and RS01 (58.97%) and RS16 (56.2%) showed the highest cholesterol assimilation rate. In addition, they also found to be resistant and sensitive to few antibiotics.Conclusion: Based on the obtained results, these isolates are ideal in vitro probiotic strains and can be used further for in vivo evaluation.


2014 ◽  
Vol 64 (Pt_11) ◽  
pp. 3862-3866 ◽  
Author(s):  
Shi Peng ◽  
Dong Dan Hong ◽  
Yang Bing Xin ◽  
Li Ming Jun ◽  
Wei Ge Hong

A Gram-staining-negative, non-motile, catalase- and oxidase-positive strain, designated CCNWSP36-1T, was isolated from the nodule surface of soybean [Glycine max (L.) Merrill] cultivar Zhonghuang 13. The 16S rRNA gene sequence analysis clearly showed that the isolate represented a member of the genus Sphingobacterium . On the basis of pairwise comparisons of 16S rRNA gene sequences, strain CCNWSP36-1T showed 96.8 % similarity to Sphingobacterium nematocida CCTCC AB 2010390T and less than 95.2 % similarity to other members of the genus Sphingobacterium . Growth of strain CCNWSP36-1T occurred at 10–40 °C and at pH 5.0–9.0. The NaCl range (w/v) for growth was 0–4 %. The predominant isoprenoid quinone was MK-7. The polar lipids were phosphatidylethanolamine and several unidentified polar lipids. Sphingolipid was present. The major fatty acids were iso-C15 : 0 and summed feature 3 (comprising C16 : 1ω6c and/or C16 : 1ω7c). The G+C content of the genomic DNA was 41.1 mol%. As the physiological and biochemical characteristics of strain CCNWSP36-1T and the type strains of its closest phylogenetic neighbours showed clear differences, a novel species, Sphingobacterium yanglingense, is proposed. The type strain is CCNWSP36-1T ( = ACCC 19328T = JCM 30166T).


2011 ◽  
Vol 61 (12) ◽  
pp. 2974-2978 ◽  
Author(s):  
Jinxing Zhu ◽  
Xiaoli Liu ◽  
Xiuzhu Dong

Two mesophilic methanogenic strains, designated TS-2T and GHT, were isolated from sediments of Tuosu lake and Gahai lake, respectively, in the Qaidam basin, Qinghai province, China. Cells of both isolates were rods (about 0.3–0.5×2–5 µm) with blunt rounded ends and Gram-staining-positive. Strain TS-2T was motile with one or two polar flagella and used only H2/CO2 for growth and methanogenesis. Strain GHT was non-motile, used both H2/CO2 and formate and displayed a variable cell arrangement depending on the substrate: long chains when growing in formate (50 mM) or under high pressure H2 and single cells under low pressure H2. Phylogenetic analysis based on 16S rRNA gene sequences placed the two isolates in the genus Methanobacterium. Strain TS-2T was most closely related to Methanobacterium alcaliphilum NBRC 105226T (96 % 16S rRNA gene sequence similarity). Phylogenetic analysis based on the alpha subunit of methyl-coenzyme M reductase also supported the affiliation of the two isolates with the genus Methanobacterium. DNA–DNA relatedness between the isolates and M. alcaliphilum DSM 3387T was 39–53 %. Hence we propose two novel species, Methanobacterium movens sp. nov. (type strain TS-2T = AS 1.5093T = JCM 15415T) and Methanobacterium flexile sp. nov. (type strain GHT = AS 1.5092T = JCM 15416T).


2011 ◽  
Vol 61 (6) ◽  
pp. 1413-1417 ◽  
Author(s):  
Yochan Joung ◽  
Kiseong Joh

A Gram-staining-negative, non-motile, yellow-pigmented bacterial strain, designated HMD1043T, was isolated from a mesotrophic artificial lake in Korea. The major fatty acids were anteiso-C15 : 0 (28.3 %), iso-C15 : 0 (22.9 %), summed feature 9 (comprising iso-C17 : 1ω9c and/or 10-methyl C16 : 0; 8.8 %) and iso-C13 : 0 (5.3 %). The DNA G+C content was 31.3 mol%. A phylogenetic tree based on 16S rRNA gene sequences showed that strain HMD1043T formed a lineage within the genus Chryseobacterium and was most closely related to Chryseobacterium antarcticum AT1013T (96.8 % 16S rRNA gene sequence similarity) and Chryseobacterium jeonii AT1047T (96.4 %). On the basis of the evidence presented in this study, strain HMD1043T is described as belonging to a novel species of the genus Chryseobacterium, for which the name Chryseobacterium yonginense sp. nov. is proposed. The type strain is HMD1043T ( = KCTC 22744T  = CECT 7547T).


2020 ◽  
Vol 10 (13) ◽  
pp. 4593 ◽  
Author(s):  
Mahshid Heidarrezaei ◽  
Hoofar Shokravi ◽  
Fahrul Huyop ◽  
Seyed Saeid Rahimian Koloor ◽  
Michal Petrů

Halogenated compounds are an important class of environmental pollutants that are widely used in industrial chemicals such as solvents, herbicides, and pesticides. Many studies have been carried out to explore the biodegradation of these chemicals. Trichloroacetic acid (TCA) is one of the main halogenated compounds that are carcinogenic to humans and animals. The bacterium was isolated from the northern coastline of Johor Strait. In this study, the ability of strain MH2 to biodegrade TCA was evaluated by a growth experiment and dehalogenase enzyme assay. The growth profile of the isolated strain was examined. The doubling time for L. boronitolerans MH2 was found to be 32 h. The release of chloride ion in the degradation process was measured at 0.33 × 10−3 ± 0.03 mol∙L−1 after 96 h when the growth curve had reached its maximum within the late bacterial exponential phase. The results showed that the strain had a promising ability to degrade TCA by producing dehalogenase enzyme when cell-free extracts were prepared from growth on TCA as the sole carbon source with enzyme-specific activity, 1.1 ± 0.05 µmolCl−min−1∙mg−1 protein. Furthermore, the morphological, and biochemical aspects of the isolated bacterium were studied to identify and characterize the strain. The morphological observation of the isolated bacterium was seen to be a rod-shaped, Gram-positive, motile, heterotrophic, and spore-forming bacterium. The amplification of the 16S rRNA and gene analysis results indicated that the isolated bacterium had 98% similarity to Lysinibacillus boronitolerans. The morphological and biochemical tests supported the 16S rRNA gene amplification. To the best of the authors’ knowledge, this is the first reported case of this genus of bacteria to degrade this type of halogenated compound.


2015 ◽  
Vol 77 (31) ◽  
Author(s):  
Suganthi Thevarajoo ◽  
Chitra Selvaratnam ◽  
Kian Mau Goh ◽  
Fazilah Abd. Manan ◽  
Zaharah Ibrahim ◽  
...  

Marine environment remained as largely unexplored source for researchers to discover marine microorganisms with novel properties. This study aims to isolate marine bacteria from the seashore of Desaru, Malaysia. Totally, six bacterial strains were successfully obtained and were identified by complete 16S rRNA sequencing. The characterizations of bacterial strains were performed based on morphological tests, Gram-staining, biochemical tests, and antibiotic sensitivity. The 16S rRNA sequence of D-2, D-4, D-7, D-15, D-31, and D-33 revealed a high identity of 97 to 99% with taxa belong to genera of Pseudomonas, Marinomonas, Exiquobacterium, Micrococcus, Pseudoalteromonas, and Shewanella respectively. Strain D-31 exhibited higher tolerance towards antibiotics kanamycin, ampicillin, and erythromycin while the growth of other strains were retarded by at least two of these antibiotics. We further characterized strain D-4 and D-31 that belonged to Marinomonas sp. and Pseudoalteromonas sp.. Both genera are interesting as earlier researchers have discovered new antibacterial substances, industrial enzymes and unique secondary metabolites.


2005 ◽  
Vol 71 (10) ◽  
pp. 5908-5919 ◽  
Author(s):  
Frederic Gich ◽  
Karin Schubert ◽  
Alke Bruns ◽  
Herbert Hoffelner ◽  
Jörg Overmann

ABSTRACT High-throughput cultivation was combined with rapid and group-specific phylogenetic fingerprinting in order to recover representatives of three freshwater bacterioplankton communities. A total of 570 bacterial cultures were obtained by employing the most probable number and MicroDrop techniques. The majority of the cultured bacteria were closely related to previously uncultured bacteria and grouped with the α-Proteobacteria, β-Proteobacteria, Actinobacteria, Firmicutes, or Flavobacteria-Cytophaga lineage. Correspondingly, the natural bacterioplankton community was analyzed by high-resolution phylogenetic fingerprinting of these five bacterial lineages. 16S rRNA gene fragments were generated for each lineage and subsequently separated by denaturing gradient gel electrophoresis. By the combination of five group-specific PCR protocols, the total number of 16S rRNA gene fingerprints generated from the natural communities was increased sixfold compared to conventional (eubacterial) fingerprinting. Four of the environmental α-Proteobacteria 16S rRNA gene sequences obtained from the natural community were found to be identical to those of bacterial isolates. One of these phylotypes was detected in 14 different cultures and hence represented the most frequently cultured bacterium. Three of these 14 strains were characterized in detail. Their complete 16S rRNA gene sequences showed only 93% similarity to that of Sandaracinobacter sibiricus, the closest relative described so far. The novel phylotype of bacterium is a strict aerobe capable of using numerous organic carbon substrates and contains bacteriochlorophyll a bound to two different photosynthetic light-harvesting complexes. Dot blot hybridization revealed that the strains occur in lakes of different trophic status and constitute up to 2% of the microbial community.


2014 ◽  
Vol 64 (Pt_11) ◽  
pp. 3768-3774 ◽  
Author(s):  
Naysim LO ◽  
Hyo Jung Kang ◽  
Che Ok Jeon

A Gram-staining-negative, facultatively aerobic bacterium, designated SM-2T, was isolated from a sea-tidal flat of Yellow Sea, South Korea. Cells were catalase- and oxidase-positive motile rods with a single polar flagellum. Growth of strain SM-2T was observed at 10–37 °C (optimum, 25–30 °C), at pH 5.5–8.5 (optimum, pH 7.0–7.5) and in the presence of 0–11 % (w/v) NaCl (optimum, 2 %). Strain SM-2T contained ubiquinone-8 (Q-8) as the sole isoprenoid quinone and C17 : 1ω8c, summed feature 3 (comprising C16 : 1ω7c and/or iso-C15 : 0 2-OH), C17 : 0 and C18 : 1ω7c as the major fatty acids. Phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unidentified lipid were identified as the major cellular polar lipids. The G+C content of the genomic DNA was 52.2 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain SM-2T formed a tight phyletic lineage with Zhongshania antarctica ZS5-23T, Zhongshania guokunii ZS6-22T and Spongiibacter borealis CL-AS9T, but that S. borealis CL-AS9T was distinct from other species of the genus Spongiibacter . Based on 16S rRNA gene sequence similarities, strain SM-2T was most closely related to S. borealis CL-AS9T, Z. antarctica ZS5-23T and Z. guokunii ZS6-22T, with similarities of 99.5 %, 98.9 % and 98.7 %, respectively, but the DNA–DNA hybridization values among these species were clearly lower than 70 %. On the basis of chemotaxonomic data and molecular properties, we propose strain SM-2T represents a novel species of the genus Zhongshania with the name Zhongshania aliphaticivorans sp. nov. (type strain SM-2T = KACC 18120T = JCM 30138T). We also propose the transfer of Spongiibacter borealis Jang et al. 2011 to the genus Zhongshania as Zhongshania borealis comb. nov. (type strain CL-AS9T = KCCM 90094T = JCM 17304T).


2010 ◽  
Vol 60 (2) ◽  
pp. 382-386 ◽  
Author(s):  
Byoung-Jun Yoon ◽  
Dong-Heon Lee ◽  
Bong-Jo Kang ◽  
Hyung-Yeel Kahng ◽  
You-Sung Oh ◽  
...  

A novel marine, Gram-staining-negative, yellow-pigmented, rod-shaped bacterial strain, designated CNU004T, was isolated from a seawater sample collected on the coastline of Jeju Island, South Korea. The strain was strictly aerobic, non-flagellated, non-gliding and oxidase- and catalase-positive. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CNU004T belongs to a distinct lineage in the family Flavobacteriaceae. Strain CNU004T exhibited levels of 16S rRNA gene sequence similarity of 93.8–93.9 % to its nearest phylogenetic neighbours, members of the genera Gaetbulibacter, Yeosuana and Algibacter. The new isolate required sea salts or artificial seawater for growth. The optimum ranges of temperature and pH for growth were 30–35 °C and pH 7.0–8.0. The DNA G+C content of strain CNU004T was 37.7 mol%. The major fatty acids were iso-C15 : 0, iso-C15 : 1 G, iso-C17 : 0 3-OH and iso-C15 : 0 3-OH. Menaquinone-6 was the major respiratory quinone. Zeaxanthin was the major carotenoid pigment produced, and flexirubin-type pigments were not produced. Strain CNU004T was able to degrade starch and agar. Based on its phenotypic and genotypic characteristics and on the phylogenetic evidence presented, strain CNU004T is considered to represent a novel species of a new genus in the family Flavobacteriaceae, for which the name Hyunsoonleella jejuensis gen. nov., sp. nov. is proposed. The type strain of Hyunsoonleella jejuensis sp. nov. is CNU004T (=KCTC 22242T =DSM 21035T).


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