Solar Saltern Research Articles

Marinobacter orientalis sp. nov., a thiosulfate-oxidizing bacterium isolated from a marine solar saltern.

A facultatively anaerobic bacterium, strain W62T, was isolated from the marine solar saltern in Weihai, China. Cells of the novel strain were Gram-stain negative, non-flagellated, non-gliding, rod-shaped and around 0.3-0.5 × 2.5-3.9µm in size. Optimum growth occurred at 33-37°C, with 3-5% (w/v) NaCl and at pH 7.0-7.5. On the basis of phylogenetic analysis of the 16S rRNA gene sequence, strain W62T had close relationship with Marinobacter vulgaris F01T (98.6%), Marinobacter confluentis KCTC 42705T (98.4%) and Marinobacter halotolerans NBRC 110910T (97.7%). Genome sequencing revealed a genome size of 4,050,555bp, a G+C content of 57.3% and a complete sox system related to thiosulfate oxidization. Strain W62T had ubiquinone-9 as the sole respiratory quinone and possessed Summed Features 3 (C16:1 ω7c/C16:1 ω6c), C16:0 and C18:1 ω9c as the major fatty acids. The major polar lipids of strain W62T were identified as aminophospholipid, phosphatidylglycerol and phosphatidylethanolamine. According to the results of the phenotypic, chemotaxonomic characterization, phylogenetic properties and genome analysis, strain W62T should represent a novel specie of the genus Marinobacter, for which the name Marinobacter orientalis sp. nov. is proposed. The type strain is W62T (= MCCC 1H00317T = KCTC 62593T).

Ecological and Metabolic Thresholds in the Bacterial, Protist, and Fungal Microbiome of Ephemeral Saline Lakes (Monegros Desert, Spain).

We studied the 16S and 18S rRNA genes of the bacterial, protist, and fungal microbiomes of 131 samples collected in 14 ephemeral small inland lakes located in the endorheic area of the Monegros Desert (NE Spain). The sampling covered different temporal flooding/desiccation cycles that created natural salinity gradients between 0.1% (w/v) and salt saturation. We aimed to test the hypothesis of a lack of competitive advantage for microorganisms using the "salt-in" strategy in highly fluctuating hypersaline environments where temperature and salinity transitions widely vary within short time periods, as in ephemeral inland lakes. Overall, 5653 bacterial zOTUs and 2658 eukaryal zOTUs were detected heterogeneously distributed with significant variations on taxonomy and general energy-yielding metabolisms and trophic strategies along the gradient. We observed a more diverse bacterial assembly than initially expected at extreme salinities and a lack of dominance of a few "salt-in" organisms. Microbial thresholds were unveiled for these highly fluctuating hypersaline environments with high selective pressures. We conclude that the extremely high dynamism observed in the ephemeral lakes of Monegros may have given a competitive advantage for more versatile ("salt-out") organisms compared to those better adapted to stable high salinities usually more common in solar salterns. Ephemeral inland saline lakes offered a well-suited natural framework for highly detailed evolutionary and ecological studies.

Rhodohalobacter mucosus sp. nov., isolated from a marine solar saltern.

A Gram-stain-negative, non-motile, aerobic, rod-shaped and reddish-pigmented bacterium, designated 8A47T, was isolated from a marine solar saltern located in Wendeng, PR China. The novel strain 8A47T grows at 20-42°C, pH 7.0-9.0, and in the presence of 2.0-14.0% (w/v) NaCl. Optimal growth was observed at 37-40°C, pH 7.5-8.0, and with 4.0-6.0% (w/v) NaCl. The phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 8A47T formed an evolutionary lineage with members of the genus Rhodohalobacter. Strain 8A47T exhibited high level of similarity to Rhodohalobacter barlenses MCCC 1K03442T (94.7%) and Rhodohalobacter halophilus JZ3C29T (93.5%). The major fatty acids were summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), C16:0 and iso-C15:0. The sole respiratory quinone was MK-7. The polar lipids of the new isolate consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, one unidentified lipid, two unidentified phospholipids and three unidentified glycolipids and. The genomic DNA G + C content of the strain 8A47T was 47.7mol%. Based on its phenotypic, chemotaxonomic, genotypic and genomic characteristics presented in this study, strain 8A47T is considered to represent a novel species of the genus Rhodohalobacter, for which the name Rhodohalobacter mucosus sp. nov. is proposed. The type strain is 8A47T (= KCTC 62603T = MCCC 1H00329T).

ÇEŞİTLİ SUCUL HİPERSALİN HABİTATLARDAKİ (TÜRKİYE) PROKARYOTİK TOPLULUKLARIN VE NANOHALOARKEAL SOYLARIN ÇEŞİTLİLİĞİNİN YÜKSEK-VERİMLİ DİZİLEME VE KLONLAMA KULLANILARAK DEĞERLENDİRİLMESİ

IIn this study, distribution of the prokaryotic groups in Tuz Lake, Ayvalık and Tuzlagözü solar salterns was investigated using 16S rRNA gene targeted approaches. The relative abundance of prokaryotic taxa in the samples was detected by using high-throughput sequencing. Operational taxonomic units (OTUs) associated with Haloquadratum were the most abundant in MiSeq reads. Nanohaloarchaeota-related OTUs were rare (<1%) in Ayvalık and Tuzlagözü solar salterns, and around 5% in Tuz Lake. Other OTUs frequently found and shared in the samples were associated with Halorubrum, Halonotius and Salinibacter genera. Nanohaloarchaeota lineages, whose phylotypes have been frequently detected in diverse hypersaline environments, were examined in more detail by 16S rRNA gene cloning using group-specific primer. Some of the highly represented nanohaloarchaeal phylotypes in the clone libraries showed low similarity to any sequence in the database, generating two distinct clades. One of the novel lineages was found to be prominent in the clone library constructed from Ayvalık sample. Phylotypes showing 95-97% sequence similarity to Ca. Nanopetramus were also highly represented in Ayvalık. Phylotypes frequent in the clone libraries of Tuz Lake and Tuzlagözü samples were associated with a novel lineage, as well as Ca. Nanosalina and its relatives.

Mesobacillus aurantius sp. nov., isolated from an orange-colored pond near a solar saltern.

An endospore producing, strict aerobic, Gram-stain-positive, orange-colored colony forming bacterium designated as strain JC1013T was isolated from an orange pond near a solar saltern of Tamil Nadu, India. Phylogenetic analysis of the 16S rRNA gene sequences indicated that strain was affiliated to the family Bacillaceae of the phylum Firmicutes. Strain showed highest 16S rRNA gene sequence identity of 98.7% with Mesobacillus selenatarsenatis SF-1T and below 98.3% with other members of the genus Mesobacillus. Strain JC1013T produced carotenoid pigments and indole compounds. Major cellular fatty acids of strain JC1013T were iso-C15:0, anteiso-C15:0, C16:0 3-OH, iso-C17:0ω10c and summed feature 4 (iso-C17:1 I/ anteisoB). Polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unidentified aminolipids and four unidentified phospholipids. Strain JC1013T constituted m-diaminopimelic acid as diagnostic cell wall amino acids. MK-7 is the predominant menaquinone of strain JC1013T. The genome size of strain JC1013T was 4.6 Mbp and its G + C content was 42.7mol%. For the affirmation of strain's taxonomic status, a detailed phylogenomic study was done. Based on the phylogenetic analyses, low ANI (84.6%), AAI (88.5%) values, in-silico DDH (< 29%) value, morphological, physiological and chemo-taxonomical characteristics, strain JC1013T was clearly distinguished from the nearest phylogenetic neighbor, Mesobacillus selenatarsenatis SF-1T to conclude that it is a new species of the genus Mesobacillus. We propose the name as Mesobacillus aurantius with type strain JC1013T (= NBRC 114146T = KACC 21451T).

Kushneria phosphatilytica sp. nov., a phosphate-solubilizing bacterium isolated from a solar saltern.

A Gram-stain-negative, motile, rod-shaped, non-endospore-forming, aerobic and halophilic bacterium, designated strain YCWA18T, was isolated from the sediment of Jimo-Daqiao saltern in China. This strain was able to grow at NaCl concentrations in the range 0.5-20 % (w/v) with optimum growth at 6 % (w/v) NaCl. Growth occurred at temperatures of 4-40 °C (optimum 28 °C) and pH 4.0-9.0 (optimum 7.0). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YCWA18T belonged to the genus Kushneria and shared the highest sequence similarity of 98.7 % with Kushneria sinocarnis DSM 23229T. Moreover, the phylogenetic analysis based on the 23S rRNA gene sequence also confirmed the phylogenetic position of this novel strain. The predominant fatty acids were C16 : 0, C17 : 0 cyclo and C12 : 0 3-OH. The major isoprenoid quinone was Q-9 (94.2 %) and the polar lipids were diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), an unidentified aminolipid (AL), an unidentified phospholipids (PL) and two unidentified lipids (L). The complete genome of strain YCWA18T consisted of a single, circular chromosome of 3 624 619 bp, with an average G+C content of 59.1 mol%. A genome-based phylogenetic tree constructed using an up-to-date bacterial core gene set (UBCG) showed that strain YCWA18T formed a clade with K. sinocarnis DSM 23229T. However, the level of the ANI and dDDH values between strain YCWA18T and K. sinocarnis DSM 23229T were 82.3 and 24.6 %, respectively, which were low enough to distinguish strain YCWA18T from K. sinocarnis DSM 23229T. Overall, based on the phenotypic, chemotaxonomic, phylogenetic and genomic analyses, strain YCWA18T represents a novel species of genus Kushneria. The name Kushneria phosphatilytica sp. nov. is proposed, with the type strain YCWA18T (=CGMCC 1.9149T=NCCB 100306T).

Shift in benthic diatom community structure and salinity thresholds in a hypersaline environment of solar saltern, Korea

The community dynamics of benthic diatoms in the hypersaline environment are investigated to advance our understanding how salinity impacts marine life. Diatoms were sampled in the two salterns encompassing salt Ponds, ditches, and seawater reservoirs (n = 11), along the salinity gradient (max = 324 psu), and nearby tidal flats (n = 2). The floral assemblages and distributions across sites and stations showed great variations, with a total of 169 identified taxa. First, not surprisingly, higher diversity of benthic diatoms was found at natural tidal flats than salterns. The saltern diatoms generally showed salinity dependent distributions with distinct spatial changes in species composition and dominant taxa. Biota-environment and principal component analysis confirmed that salinity, mud content, and total nitrogen were key factors influencing the overall benthic community structure. Some dominant species, e.g., Nitzschia scalpelliformis and Achnanthes sp. 1, showed salinity tolerance / preference. The number of diatom species at salinity of >100 psu reduced over half and no diatoms were found at maximum salinity of 324 psu. The highest salinity for the observed live diatoms was 205 psu, however, a simple regression indicated a theoretical salinity threshold of ~300 psu on the survival. Finally, the indicator species were identified along the salinity gradient in salterns as well as natural tidal flats. Overall, high species numbers, varying taxa, and euryhaline distributions of saltern diatoms collectively reflected a dynamic saltern ecosystem. The present study would provide backgrounds for biodiversity monitoring of ecologically important microalgal producers in some unique hypersaline environment, and elsewhere. Key words: benthic diatom; hypersaline; indicator species; salinity gradient; saltern

Distinct ecotypes within a natural haloarchaeal population enable adaptation to changing environmental conditions without causing population sweeps.

Microbial communities thriving in hypersaline brines of solar salterns are highly resistant and resilient to environmental changes, and salinity is a major factor that deterministically influences community structure. Here, we demonstrate that this resilience occurs even after rapid osmotic shocks caused by a threefold change in salinity (a reduction from 34 to 12% salts) leading to massive amounts of archaeal cell lysis. Specifically, our temporal metagenomic datasets identified two co-occurring ecotypes within the most dominant archaeal population of the brines Haloquadratum walsbyi that exhibited different salt concentration preferences. The dominant ecotype was generally more abundant and occurred in high-salt conditions (34%); the low abundance ecotype always co-occurred but was enriched at salinities around 20% or lower and carried unique gene content related to solute transport and gene regulation. Despite their apparent distinct ecological preferences, the ecotypes did not outcompete each other presumably due to weak functional differentiation between them. Further, the osmotic shock selected for a temporal increase in taxonomic and functional diversity at both the Hqr. walsbyi population and whole-community levels supporting the specialization-disturbance hypothesis, that is, the expectation that disturbance favors generalists. Altogether, our results provide new insights into how intraspecies diversity is maintained in light of substantial gene-content differences and major environmental perturbations.

Isolation and Taxonomic Characterization of Novel Haloarchaeal Isolates From Indian Solar Saltern: A Brief Review on Distribution of Bacteriorhodopsins and V-Type ATPases in Haloarchaea.

Haloarchaea inhabit high salinity environments worldwide. They are a potentially rich source of crucial biomolecules like carotenoids and industrially useful proteins. However, diversity in haloarchaea present in Indian high salinity environments is poorly studied. In the present study, we isolated 12 haloarchaeal strains from hypersaline Kottakuppam, Tamil Nadu solar saltern in India. 16S rRNA based taxonomic characterization of these isolates suggested that nine of them are novel strains that belong to genera Haloarcula, Halomicrobium, and Haloferax. Transmission electron microscopy suggests the polymorphic nature of these haloarchaeal isolates. Most of the haloarchaeal species are known to be high producers of carotenoids. We were able to isolate carotenoids from all these 12 isolates. The UV-Vis spectroscopy-based analysis suggests that bacterioruberin and lycopene are the major carotenoids produced by these isolates. Based on the visual inspection of the purified carotenoids, the isolates were classified into two broad categories i.e., yellow and orange, attributed to the differences in the ratio of bacterioruberin and lycopene as confirmed by the UV-Vis spectral analysis. Using a PCR-based screening assay, we were able to detect the presence of the bacteriorhodopsin gene (bop) in 11 isolates. We performed whole-genome sequencing for three bop positive and one bop negative haloarchaeal isolates. Whole-genome sequencing, followed by pan-genome analysis identified multiple unique genes involved in various biological functions. We also successfully cloned, expressed, and purified functional recombinant bacteriorhodopsin (BR) from one of the isolates using Escherichia coli as an expression host. BR has light-driven proton pumping activity resulting in the proton gradient across the membrane, which is utilized by V-Type ATPases to produce ATP. We analyzed the distribution of bop and other accessory genes involved in functional BR expression and ATP synthesis in all the representative haloarchaeal species. Our bioinformatics-based analysis of all the sequenced members of genus Haloarcula suggests that bop, if present, is usually inserted between the genes coding for B and D subunits of the V-type ATPases operon. This study provides new insights into the genomic variations in haloarchaea and reports expression of new BR variant having good expression in functional form in E. coli.

Depthwise microbiome and isotopic profiling of a moderately saline microbial mat in a solar saltern

The solar salterns in Tuticorin, India, are man-made, saline to hypersaline systems hosting some uniquely adapted populations of microorganisms and eukaryotic algae that have not been fully characterized. Two visually different microbial mats (termed ‘white’ and ‘green’) developing on the reservoir ponds (53 PSU) were isolated from the salterns. Firstly, archaeal and bacterial diversity in different vertical layers of the mats were analyzed. Culture-independent 16S rRNA gene analysis revealed that both bacteria and archaea were rich in their diversity. The top layers had a higher representation of halophilic archaea Halobacteriaceae, phylum Chloroflexi, and classes Anaerolineae, Delta- and Gamma- Proteobacteria than the deeper sections, indicating that a salinity gradient exists within the mats. Limited presence of Cyanobacteria and detection of algae-associated bacteria, such as Phycisphaerae, Phaeodactylibacter and Oceanicaulis likely implied that eukaryotic algae and other phototrophs could be the primary producers within the mat ecosystem. Secondly, predictive metabolic pathway analysis using the 16S rRNA gene data revealed that in addition to the regulatory microbial functions, methane and nitrogen metabolisms were prevalent. Finally, stable carbon and nitrogen isotopic compositions determined from both mat samples showed that the δ13Corg and δ15Norg values increased slightly with depth, ranging from − 16.42 to − 14.73‰, and 11.17 to 13.55‰, respectively. The isotopic signature along the microbial mat profile followed a pattern that is distinctive to the community composition and net metabolic activities, and comparable to saline mats in other salterns. The results and discussions presented here by merging culture-independent studies, predictive metabolic analyses and isotopic characterization, provide a collective strategy to understand the compositional and functional characteristics of microbial mats in saline environments.

Production and characterization of a bioactive extracellular homopolysaccharide produced by Haloferax sp. BKW301.

The production of extracellular polysaccharides (EPS) by haloarchaeal members, with novel and unusual physicochemical properties,is of special importance and has the potentialfor extensive biotechnological exploitation. An extremely halophilic archaeon, Haloferax sp. BKW301 (GenBank Accession No. KT240044) isolated from a solar saltern of Baksal, West Bengal, India has been optimized for the production of EPS under batch culture. It produced a considerable amount (5.95 g/L) of EPS in themedium for halophiles with 15% NaCl, 3% glucose, 0.5% yeast extract, and 6% inoculum under shake flask culture at 120 rpm. The purified EPS, a homopolymer of galactose as revealed by chromatographic methods and Fourier-transform infrared spectroscopy, is noncrystalline (CIxrd , 0.82), amorphous, and could emulsify hydrocarbons like kerosene, petrol, xylene, and so forth. Moreover, the polymer is highly thermostable (up to 420°C) and displayed pseudoplastic rheology. Biologically, the EPS was able to scavenge DPPH (2,2-diphenyl-1-picrylhydrazyl) radical efficiently and inhibit the proliferation of the Huh-7 cell line at an IC50 value of 6.25 µg/ml with a Hill coefficient of 0.844. Large-scale production of this thermostable, pseudoplastic homopolysaccharide, therefore, could find suitable applications in industry and biotechnology.

A long-term study on Coolia monotis distribution from the south-east Mediterranean Sea

The potentially toxic species Coolia monotis is widespread along the Mediterranean coasts. The Gulf of Gabès is one of the affected areas in the south-eastern part of the Mediterranean Sea. Distribution patterns of C. monotis in relation to temperature and salinity were investigated at 24 monitoring sites along the Gulf of Gabès coasts during 22 years (1997–2018). The analysis of C. monotis occurrence in the water column revealed a seasonal trend with high abundances (≥105 cells l−1) recorded during winter. C. monotis was widely distributed in Gabès Gulf, it was most abundant in the southern part, particularly at the Cotusal Channel site, which is surrounded by marine solar salterns, where the highest concentration (2.54 × 106 cells l−1), the maximum mean abundance (3.74 ± 51.87 × 104 cells l−1) and the highest number of observations (75) were recorded over the study period. C. monotis was observed within a wide range of temperature (9–31.6 °C) and salinity (31–59) with high abundances (>105 cells l−1) occurring at water salinity and temperature of 41.9–45 and 15–17.3 °C, respectively. A positive correlation was found between the abundance of C. monotis and salinity (p < 0.05), whereas no correlation was found as regards temperature. Culture experiments were conducted under varying salinity regimes (10–50) with C. monotis strain (Com.10) isolated from the Cotusal Channel. Growth occurred at salinities between 20 and 50 and the maximum growth rate of 0.23 day−1was recorded at a salinity of 40. Results suggest that seawater salinity plays a central role in the C. monotis blooms development.

Resistance of a Halobacterium salinarum isolate from a solar saltern to cadmium, lead, nickel, zinc, and copper.

The current study focuses on the tolerance of a strain of Halobacterium salinarum isolated from Sfax solar saltern (Tunisia) towards cadmium (Cd), lead (Pb), nickel (Ni), zinc (Zn), and copper (Cu) by using agar dilution methods in complex and minimal media. The results showed the least inhibitory metals based on Minimum Inhibitory Concentrations (MICs) were lead (MIC = 4.5mM), cadmium (MIC = 4mM), and nickel (MIC = 2.5mM) in complex medium. The MICs of these metals were more inhibitory (MIC < 2mM) in the other tested media. The archaeal strain revealed a high sensitivity for copper and zinc, with MICs below 0.5mM for both metals. Growth kinetics in complex and minimal media showed the strain to be more sensitive to the metals in liquid media than in solid media. The growth kinetic assays indicated the presence of selected heavy metals resulted in a lower growth rate and lower total cell mass relative to the control. Despite that cadmium and lead are nonessential and have no nutrient value, they were the most tolerated metals by H. salinarum strain. In addition, pigment intensity in the strain was inhibited by the presence of the heavy metals relative to the control.

Poorly known microbial taxa dominate the microbiome of hypersaline Sambhar Lake salterns in India.

Inland athalassohaline solar salterns provide unique opportunity to study microbial successions along salinity gradients that resemble transition in natural hypersaline lakes. We analyzed for the first time 16S rRNA gene amplicon sequences of bacteria (V1-V2) and archaea (V4-V5) in saltern brines of India's largest inland hypersaline Sambhar Lake. Brines of the salterns (S1-S4) are alkaline (pH 9.5-10.5) with salinities of 130, 170, 280 and 350 gL-1 respectively. 16S rRNA gene copy-number of archaea outnumbered that of bacteria in all salterns. Their diversity also increased along S1 through S4, while that of bacteria decreased. Brines of S3 and S4 were dominated by specialized extreme halophilic bacterial (Halanaerobiales, Rhodothermaceae) and archaeal (Halobacteriales, Haloferacales) members with recognized salt-in strategy for osmoadaptation. Microbial assemblages positively correlated to saltern pH, total salinity, and ionic composition. Archaea in S1 and S2 were unprecedentedly represented by poorly known as-yet uncultivated groups, Woesearchaeota (90.35-93.51%) and Nanohaloarchaeota that belong to the newly proposed nano-sized superphylum DPANN. In fact, these taxa were identified in archaeal datasets of other athalassohaline salterns after re-analysis using latest RDP database. Thus, microbial compositions in hypersaline lakes are complex and need revisit particularly for their archaeal diversity to understand their hitherto unknown ecological function in extreme environments.

Biochemical and Taxonomic Characterization of Novel Haloarchaeal Strains and Purification of the Recombinant Halotolerant α-Amylase Discovered in the Isolate.

Haloarchaea are salt-loving archaea and potential source of industrially relevant halotolerant enzymes. In the present study, three reddish-pink, extremely halophilic archaeal strains, namely wsp1 (wsp-water sample Pondicherry), wsp3, and wsp4, were isolated from the Indian Solar saltern. The phylogenetic analysis based on 16S rRNA gene sequences suggests that both wsp3 and wsp4 strains belong to Halogeometricum borinquense while wsp1 is closely related to Haloferax volcanii species. The comparative genomics revealed an open pangenome for both genera investigated here. Whole-genome sequence analysis revealed that these isolates have multiple copies of industrially/biotechnologically important unique genes and enzymes. Among these unique enzymes, for recombinant expression and purification, we selected four putative α-amylases identified in these three isolates. We successfully purified functional halotolerant recombinant Amy2, from wsp1 using pelB signal sequence-based secretion strategy using Escherichia coli as an expression host. This method may prove useful to produce functional haloarchaeal secretory recombinant proteins suitable for commercial or research applications. Biochemical analysis of Amy2 suggests the halotolerant nature of the enzyme having maximum enzymatic activity observed at 1 M NaCl. We also report the isolation and characterization of carotenoids purified from these isolates. This study highlights the presence of several industrially important enzymes in the haloarchaeal strains which may potentially have improved features like stability and salt tolerance suitable for industrial applications.

Phylogenetic characterization and screening of halophilic bacteria from Algerian salt lake for the production of biosurfactant and enzymes

Environments containing significant concentration of NaCl such as salt lakes harbor extremophiles microorganisms which have a great biotechnology interest. To explore the diversity of Bacteria in Chott Tinsilt (Algeria), an isolation program was performed. Water samples were collected from the saltern during the pre-salt harvesting phase. This Chott is high in salt (22.47% (w/v). Seven halophiles Bacteria were selected for further characterization. The isolated strains were able to grow optimally in media with 10–25% (w/v) total salts. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. It showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Planococcus and Halobacillus genera with less than 98% of similarity with their closest phylogenetic relative. The halophilic bacterial isolates were also characterized for the production of biosurfactant and industrially important enzymes. Most isolates produced hydrolases and biosurfactants at high salt concentration. In fact, this is the first report on bacterial strains (A4 and B4) which were a good biosurfactant and coagulase producer at 20% and 25% ((w/v)) NaCl. In addition, the biosurfactant produced by the strain B4 at high salinity (25%) was also stable at high temperature (30-100°C) and high alkalinity (pH 11).

Transient Dynamics of Archaea and Bacteria in Sediments and Brine Across a Salinity Gradient in a Solar Saltern of Goa, India.

The microbial fluctuations along an increasing salinity gradient during two different salt production phases – initial salt harvesting (ISH) phase and peak salt harvesting (PSH) phase of Siridao solar salterns in Goa, India were examined through high-throughput sequencing of 16S rRNA genes on Illumina MiSeq platform. Elemental analysis of the brine samples showed high concentration of sodium (Na+) and chloride (Cl–) ions thereby indicating its thalassohaline nature. Comparison of relative abundance of sequences revealed that Archaea transited from sediment to brine while Bacteria transited from brine to sediment with increasing salinity. Frequency of Archaea was found to be significantly enriched even in low and moderate salinity sediments with their relative sequence abundance reaching as high as 85%. Euryarchaeota was found to be the dominant archaeal phylum containing 19 and 17 genera in sediments and brine, respectively. Phylotypes belonging to Halorubrum, Haloarcula, Halorhabdus, and Haloplanus were common in both sediments and brine. Occurence of Halobacterium and Natronomonas were exclusive to sediments while Halonotius was exclusive to brine. Among sediments, relative sequence frequency of Halorubrum, and Halorhabdus decreased while Haloarcula, Haloplanus, and Natronomonas increased with increasing salinity. Similarly, the relative abundance of Haloarcula and Halorubrum increased with increasing salinity in brine. Sediments and brine samples harbored about 20 and 17 bacterial phyla, respectively. Bacteroidetes, Proteobacteria, and Chloroflexi were the common bacterial phyla in both sediments and brine while Firmicutes were dominant albeit in sediments alone. Further, Gammaproteobacteria, Alphaproteobacteria, and Deltaproteobacteria were observed to be the abundant class within the Proteobacteria. Among the bacterial genera, phylotypes belonging to Rubricoccus and Halomonas were widely detected in both brine and sediment while Thioalkalispira, Desulfovermiculus, and Marinobacter were selectively present in sediments. This study suggests that Bacteria are more susceptible to salinity fluctuations than Archaea, with many bacterial genera being compartment and phase-specific. Our study further indicated that Archaea rather than Bacteria could withstand the wide salinity fluctuation and attain a stable community structure within a short time-frame.

Symbiosis between nanohaloarchaeon and haloarchaeon is based on utilization of different polysaccharides

Nano-sized archaeota, with their small genomes and limited metabolic capabilities, are known to associate with other microbes, thereby compensating for their own auxotrophies. These diminutive and yet ubiquitous organisms thrive in hypersaline habitats that they share with haloarchaea. Here, we reveal the genetic and physiological nature of a nanohaloarchaeon-haloarchaeon association, with both microbes obtained from a solar saltern and reproducibly cultivated together in vitro. The nanohaloarchaeon Candidatus Nanohalobium constans LC1Nh is an aerotolerant, sugar-fermenting anaerobe, lacking key anabolic machinery and respiratory complexes. The nanohaloarchaeon cells are found physically connected to the chitinolytic haloarchaeon Halomicrobium sp. LC1Hm. Our experiments revealed that this haloarchaeon can hydrolyze chitin outside the cell (to produce the monosaccharide N-acetylglucosamine), using this beta-glucan to obtain carbon and energy for growth. However, LC1Hm could not metabolize either glycogen or starch (both alpha-glucans) or other polysaccharides tested. Remarkably, the nanohaloarchaeon's ability to hydrolyze glycogen and starch to glucose enabled growth of Halomicrobium sp. LC1Hm in the absence of a chitin. These findings indicated that the nanohaloarchaeon-haloarchaeon association is both mutualistic and symbiotic; in this case, each microbe relies on its partner's ability to degrade different polysaccharides. This suggests, in turn, that other nano-sized archaeota may also be beneficial for their hosts. Given that availability of carbon substrates can vary both spatially and temporarily, the susceptibility of Halomicrobium to colonization by Ca Nanohalobium can be interpreted as a strategy to maximize the long-term fitness of the host.

Habitat Ecology and Biological Characteristics of a Hypersaline Ciliate, Fabrea salina from Solar Salterns of Mumbai Coast, India

Habitat Ecology and Biological Characteristics of a Hypersaline Ciliate, Fabrea salina from Solar Salterns of Mumbai Coast, India

LEVAN PRODUCTION POTENTIALS FROM DIFFERENT HYPERSALINE ENVIRONMENTS IN TURKEY

12 halophilic strains from different hypersaline environments such as solar salterns in Tuzlagözü (Sivas), Fadlum (Sivas), Kemah (Erzincan), a hypersaline spring water in Pülümür (Tunceli) and a saline lake in Delice (Kırıkkale) belonging to Turkey, were investigated in terms of levan production. After incubation and ethyl alcohol treatment, dialysis process was operated for partial purification. Levan amounts in our samples after hydrolysis were calculated based on the amount of sugar obtained by acid hydrolysis of standard levan. Sugar amount of samples were determined using by high performance liquid chromatography system (HPLC). 1H-Nuclear Magnetic Resonance (1H-NMR) spectra of the levan sample and standard were recorded. The results obtained by HPLC analysis showed that Chromohalobacter canadensis strain 85B had highest production potential as 234.67 mg levan/g biomass. The chemical shifts of 1H-NMR spectrum of the extracted levan also showed high similarity to those of pure levan isolated from Erwinia herbicola. Furthermore, Marinobacter sp. 163Y strain also is also capable with regard to levan production. In this study, this strain could yield 230.80 mg/g levan and this potential was first reported in the literature.