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PubMed2007

Denitrification in a binary culture and thiocyanate metabolism in Thiohalophilus thiocyanoxidans gen. nov. sp. nov. - a moderately halophilic chemolithoautotrophic sulfur-oxidizing Gammaproteobacterium from hypersaline lakes.

Anaerobic enrichment culture with thiocyanate as electron donor and nitrate as electron acceptor at 2 M NaCl inoculated with a mixture of sediments from hypersaline lakes in Kulunda Steppe (Altai, Russia) resulted in a selection of a binary consortium of moderately halophilic, obligately chemolithoautotrophic sulfur-oxidizing bacteria (SOB) capable of complete denitrification of nitrate with thiosulfate as the electron donor. One consortium member, strain HRhD 3sp, was isolated into pure culture with nitrate and thiosulfate using a density gradient. This strain was responsible for the reduction of nitrate to nitrite in the consortium, while a second strain, HRhD 2, isolated under microoxic conditions with thiosulfate as substrate, was capable of anaerobic growth with nitrite and thiosulfate. Nitrite, either as substrate or as product, was already toxic at very low concentrations for both strains. As a result, optimal growth under anaerobic conditions could only be achieved within the consortium. On the basis of phylogenetic analysis, both organisms were identified as new lineages within the Gammaproteobacteria. As well as thiosulfate, strain HRhD 2 can also use thiocyanate as electron donor, representing a first halophilic SOB capable of growth with thiocyanate at 2-4 M NaCl. Product and enzymatic analysis identified the "carbonyl sulfide (COS) pathway" of primary thiocyanate degradation in this new species. On the basis of phenotypic and genetic analysis, strain HRhD 2 is proposed to be assigned to a new genus and species Thiohalophilus thiocyanoxidans.

Aerobiosis↗

PubMed2002

Effect of oxygen concentration on photosynthesis and respiration in two hypersaline microbial mats.

The effects of oxygen concentration on photosynthesis and respiration in two hypersaline cyanobacterial mats were investigated. Experiments were carried out on mats from Eilat, Israel, with moderate photosynthetic activity, and mats from Mallorca, Spain, with high photosynthetic activity. The oxygen concentration in the overlying water above the mats was increased stepwise from 0% to 100% O2. Subsequent changes in oxygen concentration, gross photosynthetic rates, and pH values inside the mats were measured with microelectrodes. According to published reports on the regulation of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), the key enzyme in the CO2-fixation pathway of phototrophs, we expected photosynthetic activity to decrease with increasing oxygen concentration. Gross photosynthetic and total respiration rates in both mats were highest when the O2 concentration was at 0% in the overlying water. Net oxygen production rates under these conditions were the same as under air saturation (21% O2), while gross photosynthetic and respiration rates were lowest at air saturation. In both mats, gross photosynthetic and respiration rates increased upon gradually increasing the oxygen concentration in the overlying water from 21% to 100%. These results contradict the expectation that photosynthesis decreases with increasing oxygen concentration. Increased photosynthetic rates at oxygen concentrations above 21% were probably caused by enhanced oxidation of organic matter and concomitant CO2 production due to the increased oxygen availability. The cause of the high respiration rates at 0% O2 in the overlying water was presumably the enhanced excretion of photosynthetic products during increased photosynthesis. We conclude that the effect of the O2/CO2 concentration ratio on the activity of Rubisco as demonstrated in vitro on enzyme extracts cannot be extrapolated to the situation in intact microbial mats, because the close coupling of the activity of primary producers and heterotrophic bacteria plays a major role in this ecosystem.

Algorithms↗

PubMed2004

Vertical distribution of nitrogen-fixing phylotypes in a meromictic, hypersaline lake.

We investigated the diversity of nitrogenase genes in the alkaline, moderately hypersaline Mono Lake, California to determine (1) whether nitrogen-fixing (diazotrophic) populations were similar to those in other aquatic environments and (2) if there was a pattern of distribution of phylotypes that reflected redox conditions, as well as (3) to identify populations that could be important in N dynamics in this nitrogen-limited lake. Mono Lake has been meromictic for almost a decade and has steep gradients in oxygen and reduced compounds that provide a wide range of aerobic and anaerobic habitats. We amplified a fragment of the nitrogenase gene (nifH) from planktonic DNA samples collected at three depths representing oxygenated surface waters, the oxycline, and anoxic, ammonium-rich deep waters. Forty-three percent of the 90 sequences grouped in nifH Cluster I. The majority of clones (57%) grouped in Cluster III, which contains many known anaerobic bacteria. Cluster I and Cluster III sequences were retrieved at every depth indicating little vertical zonation in sequence types related to the prominent gradients in oxygen and ammonia. One group in Cluster I was found most often at every depth and accounted for 29% of all the clones. These sequences formed a subcluster that contained other environmental clones, but no cultivated representatives. No significant nitrogen fixation was detected by the 15N2 method after 48 h of incubation of surface, oxycline, or deep waters, suggesting that pelagic diazotrophs were contributing little to nitrogen fluxes in the lake. The failure to measure any significant nitrogen fixation, despite the detection of diverse and novel nitrogenase genes throughout the water column, raises interesting questions about the ecological controls on diazotrophy in Mono Lake and the distribution of functional genes in the environment.

Bacteria↗

PubMed2004

Abundance, distribution, and diversity of viruses in alkaline, hypersaline Mono Lake, California.

Mono Lake is a large (180 km2), alkaline (pH approximately 10), moderately hypersaline (70-85 g kg(-1)) lake lying at the western edge of the Great Basin. An episode of persistent chemical stratification (meromixis) was initiated in 1995 and has resulted in depletion of oxygen and accumulation of ammonia and sulfide beneath the chemocline. Although previous studies have documented high bacterial abundances and marked seasonal changes in phytoplankton abundance and community composition, there have been no previous reports on the occurrence of viruses in this unique lake. Based on the high concentrations and diversity of microbial life in this lake, we hypothesized that planktonic viruses are also abundant and diverse. To examine the abundance and distribution of viruses and bacteria, water samples were collected from four stations along 5 to 15 vertical depths at each station. Viral abundance ranged from 1 x 10(8) to 1 x 10(9) mL(-1), among the highest observed in any natural aquatic system examined so far. Increases (p < 0.1) in viral densities were observed in the anoxic bottom water at multiple stations. However, regression analysis indicated that viral abundance could not be predicted by any single environmental parameter. Pulsed field gel electrophoresis revealed a diverse viral community in Mono Lake with genome sizes ranging from approximately 14 to >400 kb with most of the DNA in the 30 to 60 kb size range. Cluster analysis grouped the anoxic bottom-water viral community into a unique cluster differentiating it from surface and mid-water viral communities. A hybridization study using an indigenous viral isolate as a probe revealed an episodic pattern of temporal phage distribution with strong niche stratification between oxic and anoxic waters.

Ammonia↗

PubMed2004

Degradation of 2,4-dichlorophenoxyacetic acid (2,4-d) by a hypersaline microbial mat and related functional changes in the mat community.

Microbial mats possibly possess degradation capacities for haloorganic pollutants because of their wide range of different functional groups of microorganisms combined with extreme diurnal changes in pH, oxygen, and sulfide gradients. In this study, 20 mg/l of the chlorinated herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) was applied to a pristine hypersaline cyanobacterial mat from Guerrero Negro, Mexico, under a light regime of 12 h dark/12 h light (600 mumol photons/m(2)s). The loss of 2,4-D was followed by chemical GC analysis; functional changes within the mat were determined with microelectrodes for oxygen, photosynthesis, pH, and sulfide. The depletion of 2,4-D due to photooxidation or sorption processes was checked in control experiments. Within 13 days, the light/dark incubated mats degraded 97% of the herbicide, while in permanent darkness only 35% were degraded. Adsorption of 2,4-D to the mat material, agar, or glass walls was negligible (4.6%), whereas 21% of the herbicide was degraded photochemically. The 2,4-D removal rate in the light/dark incubations was comparable to values reported for soils. The phototrophic community of the mat was permanently inhibited by the 2,4-D addition by 17% on average. The sulfate reduction in the entire mat and the respiration in the photic zone were inhibited more strongly but returned to original levels. Since at the end of the experiment the photosynthetic and respiratory activity of the mats were almost as high as in the beginning and 2,4-D almost completely disappeared, we conclude that the examined mats represent a robust and effective system for the degradation of the herbicide where probably the aerobic heterotrophic population is a major player in the degradation process.

2,4-Dichlorophenoxyacetic Acid↗

PubMed2005

Aerobic organic carbon mineralization by sulfate-reducing bacteria in the oxygen-saturated photic zone of a hypersaline microbial mat.

The sulfate-reducing bacterium strain SRB D2 isolated from the photic zone of a hypersaline microbial mat, from Lake Chiprana, NE Spain, respired pyruvate, alanine, and alpha-ketoglutarate but not formate, lactate, malate, succinate, and serine at significant rates under fully oxic conditions. Dehydrogenase enzymes of only the former substrates are likely oxygen-tolerant as all substrates supported anaerobic sulfate reduction. No indications were found, however, that aerobic respiration supported growth. Although strain SRB D2 appeared phylogenetically closely related to the oxygen-tolerant sulfate-reducing bacterium Desulfovibrio oxyclinae, substrate spectra were markedly different. Most-probable-number (MPN) estimates of sulfate-reducing bacteria and aerobic heterotrophic bacteria indicated that the latter were numerically dominant in both the photic and aphotic zones of the mat. Moreover, substrate spectra of representative isolates showed that the aerobic heterotrophic bacteria are metabolically more diverse. These findings indicate that sulfate-reducing bacteria in the fully oxic photic zone of mats have to compete with aerobic heterotrophic bacteria for organic substrates. Porewater analysis revealed that total carbohydrates and low-molecular-weight carbon compounds (LMWC) made up substantial fractions of the total dissolved organic carbon (DOC) pool and that nighttime degradation of the former was concomitant with increased concentration of the latter. Our findings indicate that aerobic respiration by sulfate-reducing bacteria contributes to organic carbon mineralization in the oxic zone of microbial mats as daytime porewater LMWC concentrations are above typical half-saturation constants.

Aerobiosis↗

PubMed2006

Screening and identification of endomannanase-producing microfungi from hypersaline environments.

A culture-dependent enrichment technique was used to isolate endo-1,4-beta-mannanase-producing fungi from a hypersaline environment. Galactomannan was used as carbon source and resulted in isolation of strains of Scopulariopsis brevicaulis, S. candida, and Verticillium dahliae. The Scopulariopsis isolates were found to be more dominant and could be isolated from consecutive evaporation ponds, whereas Verticillium was only isolated from one pond. The Scopulariopsis strains exhibited only endomannanase activity, whereas Verticillium displayed broad-activity spectrum by secreting endoxylanases and cellulases in addition to endomannanases. S. candida LMK004 and LMK008 produced 7,420 and 14,750 nkat g(-1) biomass, respectively. Endomannanase production in these strains increased with an increase in NaCl concentration up to 10% (w/v), after which both growth and enzyme production was decreased. V. dahliae LMK006 grew and produced up to 5,000 nkat g(-1) biomass endomannanase in the absence of NaCl. Increased NaCl concentration had a negative effect on this strain. S. brevicaulis LMK002 showed poor endomannanase production but a similar growth trend as the other Scopulariopsis strains. In general, the Scopulariopsis strains exhibited better halotolerance than V. dahliae and could grow in the presence of 20% NaCl on solid medium.

Ascomycota↗

PubMed2003

Photosynthetic performance and resource utilization of two mangrove species coexisting in a hypersaline scrub forest.

In a hypersaline mangrove scrub forest in northern Florida, coexisting trees of Laguncularia racemosa and Avicennia germinans were either fertilized with nitrogen or phosphorus, or not fertilized (controls). We aimed to test whether nutrient additions differentially altered photosynthetic performance and resource utilization in these two species. In control trees, photosynthetic rates were higher in L. racemosa than A. germinans. However, leaf nitrogen concentrations were higher in A. germinans than L. racemosa. Avicennia germinans responded to fertilization with nitrogen by increasing leaf nitrogen concentrations and rates of photosynthesis such that they were equivalent to photosynthesis in L. racemosa. Laguncularia racemosa did not show a response to nitrogen additions. Neither species showed strong responses to phosphorus fertilization. Avicennia germinans had high photosynthetic water-use efficiency (photosynthesis/transpiration), but low photosynthetic nitrogen-use efficiency (photosynthesis/leaf nitrogen). In contrast, L. racemosa had comparatively low photosynthetic water use efficiency and high photosynthetic nitrogen use efficiency. Leaf level characteristics lead us to hypothesize that coexistence of A. germinans and L. racemosa should occur where nitrogen levels are low and salinity is moderate, or at least moderate for some period of the year.

Avicennia↗

PubMed2004

Thialkalivibrio halophilus sp. nov., a novel obligately chemolithoautotrophic, facultatively alkaliphilic, and extremely salt-tolerant, sulfur-oxidizing bacterium from a hypersaline alkaline lake.

A new chemolithoautotrophic, facultatively alkaliphilic, extremely salt-tolerant, sulfur-oxidizing bacterium was isolated from an alkaline hypersaline lake in the Altai Steppe (Siberia, Russia). According to 16S rDNA analysis and DNA-DNA hybridization, strain HL 17T was identified as a new species of the genus Thialkalivibrio belonging to the gamma subdivision of the Proteobacteria for which the name Thialkalivibrio halophilus is proposed. Strain HL 17T is an extremely salt-tolerant bacterium growing at sodium concentrations between 0.2 and 5 M, with an optimum of 2 M Na+. It grew at high concentrations of NaCl and of Na2CO3/NaHCO3 (soda). Strain HL 17T is a facultative alkaliphile growing at pH range 7.5-9.8, with a broad optimum between pH 8.0 and 9.0. It used reduced inorganic sulfur compounds (thiosulfate, sulfide, polysulfide, elemental sulfur, and tetrathionate) as energy sources and electron donors. In continuous culture under energy limitation, thiosulfate was stoichiometrically oxidized to sulfate. In sodium carbonate medium under alkaline conditions, the maximum growth rate was similar, while the biomass yield was lower as compared with the NaCl-grown culture. The maximum sulfur-oxidizing capacity measured in washed cells was higher in the soda buffer independent of the growth conditions. The compatible solute content of the biomass was higher in the sodium chloride-grown culture than in the sodium carbonate/bicarbonate-grown culture. The data suggest that the osmotic pressure differences between soda and NaCl solutions might be responsible for the difference observed in compatible solutes production. This may have important implications in overall energetic metabolism of high salt adaptation.

Chromatiaceae↗

PubMed2005

Oxidation of thiosulfate to tetrathionate by an haloarchaeon isolated from hypersaline habitat.

A novel, extremely halophilic, neutrophilic archaeon was isolated from a mixed sediment sample from different hypersaline lakes in Kulunda steppe (Altai, Russia) at 4 M NaCl with acetate and thiosulfate as substrates. The enrichment culture developed in two phases. During the first phase, a rapid growth of heterotrophic, red-colored, polymorphic rods occurred with the concomitant oxidation of thiosulfate to tetrathionate. The latter was subsequently oxidized to sulfate during a second, slower phase by extremely halophilic, chemolithoautotrophic bacteria belonging to the gamma subdivision of the Proteobacteria. The archaeal strain HG 1 was isolated from the first phase of the enrichment culture using acetate as substrate. It was able to oxidize thiosulfate to tetrathionate during heterotrophic growth with acetate-a property not yet demonstrated for any of the known haloarchaea. The presence of tetrathionate synthase, the enzyme responsible for thiosulfate oxidation, was detected in strain HG 1. The activity was associated with membranes and depended specifically on Cl-, in contrast to the similar activity in extremely halophilic sulfur-oxidizing Gammaproteobacteria from the same enrichment, which was soluble and demanded both Na+ and Cl- . Strain HG 1 was identified as a member of the genus Natronorubrum.

Archaea↗

PubMed2006

Influence of recirculation on the performance of anaerobic sequencing batch biofilm reactor (AnSBBR) treating hypersaline composite chemical wastewater.

Influence of recirculation on the performance of anaerobic sequencing batch biofilm reactor (AnSBBR) was studied in the process of treating hypersaline (total dissolved inorganic solids (TDIS) approximately 26 g/l) and low biodegradable (BOD/COD approximately 0.3) composite chemical wastewater. Significant enhancement in the substrate removal efficiency and biogas yield was observed after introducing the recirculation to the system. Maximum efficiency (COD removal efficiency - 51%; SDR - 3.14 kg COD/cum-day) was observed at recirculation to feed (R/F) ratio of 2 (OLR - 6.15 kg C OD/cum-day; HLR - 2.30 cum (liquid)/cum day; UFV(A) - 0.023 m/h). Subsequent increase of R/F to 3 (OLR - 6.15 kg COD/cum-day; HLR - 3.07cum (liquid)/cum-day; UFV(A) - 0.035 m/h) resulted in reduction in COD removal efficiency (32%; SDR - 1.97 kg COD/cum-day). The enhanced performance of the system due to the introduction of recirculation was attributed to the improvement in the mass transfer between the substrate present in the bulk liquid and the attached biofilm. The hydrodynamic behavior due to recirculation mode of operation reduced the concentration gradient (substrate inhibition) of substrate and reaction by-products (VFA) resulting in mixed flow conditions.

Bacteria, Anaerobic↗

PubMed2005

Microbial enzymes mined from the Urania deep-sea hypersaline anoxic basin.

We created a metagenome expression library from the brine:seawater interface of the Urania hypersaline basin, screened it for esterases, and characterized five of these. Two had no significant sequence homology to known esterases, hydrolyzed both carboxylesters and thioesters, and exhibited unusual, habitat-specific characteristics (preference for high hydrostatic pressure and salinity). One has an unusual structural signature incorporating three catalytic active centers mediating distinct hydrolytic activities and an adaptive tertiary-quaternary structure that alters between three molecular states, according to the prevailing physicochemical conditions. Some of the esterases have high activities, specificities, enantioselectivities, and exceptional stability in polar solvents, and they are therefore potentially useful for industrial biotransformations. One possesses the highest enantioselectivity toward an ester of the important chiral synthon solketal (E: 126[S]; 98%ee).

Bacteria, Anaerobic↗

PubMed2005

Mathematical simulation of the diel O, S, and C biogeochemistry of a hypersaline microbial mat.

The creation of a mathematical simulation model of photosynthetic microbial mats is important to our understanding of key biogeochemical cycles that may have altered the atmospheres and lithospheres of early Earth. A model is presented here as a tool to integrate empirical results from research on hypersaline mats from Baja California Sur (BCS), Mexico into a computational system that can be used to simulate biospheric inputs of trace gases to the atmosphere. The first version of our model, presented here, calculates fluxes and cycling of O(2), sulfide, and dissolved inorganic carbon (DIC) via abiotic components and via four major microbial guilds: cyanobacteria (CYA), sulfate reducing bacteria (SRB), purple sulfur bacteria (PSB) and colorless sulfur bacteria (CSB). We used generalized Monod-type equations that incorporate substrate and energy limits upon maximum rates of metabolic processes such as photosynthesis and sulfate reduction. We ran a simulation using temperature and irradiance inputs from data collected from a microbial mat in Guerrero Negro in BCS (Mexico). Model O(2), sulfide, and DIC concentration profiles and fluxes compared well with data collected in the field mats. There were some model-predicted features of biogeochemical cycling not observed in our actual measurements. For instance, large influxes and effluxes of DIC across the MBGC mat boundary may reveal previously unrecognized, but real, in situ limits on rates of biogeochemical processes. Some of the short-term variation in field-collected mat O(2) was not predicted by MBGC. This suggests a need both for more model sensitivity to small environmental fluctuations for the incorporation of a photorespiration function into the model.

Carbon↗

PubMed2005

Aerobic biodegradation of benzene and toluene under hypersaline conditions at the Great Salt Plains, Oklahoma.

The Great Salt Plains is a 65-km(2) hypersaline habitat of geological origin located in north-central Oklahoma. Contamination of such ecosystems by petroleum compounds is expected from non-point sources and due to increased human activities. Little information exists about the ability of halophilic and halotolerant bacteria present in such ancient and uncontaminated environments to degrade aromatic hydrocarbons. An enrichment culture was established from soil samples obtained from the salt flats using benzene as the sole carbon and energy source. The enrichment degraded benzene at varied salt concentrations ranging from 0 to 4M. Studies showed that roughly 33% of the (14)C-benzene was converted to (14)CO(2), indicating the mineralization capacity of native bacteria. Bacterial community structure analysis using denaturing gradient gel electrophoresis showed that different phylotypes were dominant at different salt concentrations.

Aerobiosis↗

PubMed2006

Alkalibacillus halophilus sp. nov., a new halophilic species isolated from hypersaline soil in Xin-Jiang province, China.

A halophilic, Gram-positive, spore-forming motile Bacillus-like strain YIM 012(T), was isolated from one of the hypersaline soil samples collected in Xin-jiang province, China. Its optimum growth occurred at 10-20% of NaCl concentration (w/v), pH 7.0-8.0. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YIM 012(T) is a member of the genus of Alkalibacillus, which is well supported by its chemotaxonomic and molecular characteristics. Based on its phenotypic evidence and genotypic data, Alkalibacillus halophilus sp. nov. was proposed and strain YIM 012(T) (=DSM 17369(T)=KCTC 3990(T)) was assigned as the type strain of the novel species.

Bacillus↗

PubMed2005

Microbial lithification in marine stromatolites and hypersaline mats.

Lithification in microbial ecosystems occurs when precipitation of minerals outweighs dissolution. Although the formation of various minerals can result from microbial metabolism, carbonate precipitation is possibly the most important process that impacts global carbon cycling. Recent investigations have produced models for stromatolite formation in open marine environments and lithification in shallow hypersaline lakes, which could be highly relevant for interpreting the rock record and searching for extraterrestrial life. Two factors that are controlled by microbial processes and physicochemical characteristics determine precipitation: exopolymeric substances and the saturation index, the latter being determined by the pH, {Ca(2+)} and {CO(3)(2-)}. Here, we evaluate community metabolism in microbial mats and hypothesize why these organosedimentary biofilms sometimes lithify and sometimes do not.

Bacteria↗

PubMed2005

SH1: A novel, spherical halovirus isolated from an Australian hypersaline lake.

A novel halovirus, SH1, with a spherical morphology is described. Isolated from a hypersaline lake, SH1 is divalent, producing clear plaques on Haloarcula hispanica and a natural Halorubrum isolate. Single-step growth curves gave a latent period of 5-6 h and a burst size of around 200 PFU/cell. The host can differentiate to form tight clusters of thick cell-walled forms, and these were shown to be resistant to infection. Purified virions had no visible tail, were about 70 nm in diameter, and displayed a fragile outer capsid layer, possibly with an underlying membrane component. The structural proteins of the virion were analyzed by SDS-PAGE and several were found to be cross-linked, forming protein complexes. The genome was linear, dsDNA, of approximately 30 kb in length. This morphology and linear genome are features not observed in any other euryarchaeal viruses, but have properties similar to the bacterial virus PRD1.

Archaeal Viruses↗

PubMed2002

Mercury and lead tolerance in hypersaline sulfate-reducing bacteria.

Sulfate-reducing bacteria (SRB) HSR1, HSR4, and HSR14 isolated from the salt pans of Goa grew best at 90-100/1000 salinity on substrates like formate, acetate, lactate, butyrate, ethanol and benzoate. They were gram negative, non-sporulating, non-motile rods lacking in desulfoviridin and cytochromes. Examination of these isolates for heavy metal tolerance and response studies in terms of growth and sulfate-reducing activity (SRA) were carried out using HgCl2 and Pb(NO3)2 at final concentration of 50, 100, and 200 and 100, 200 and 500 microg ml(-1) respectively. With Hg, HSR1 showed approximately 80% of the control's growth at 100 and 200 microg ml(-1) but SRA reached only 60% of the control values at the end of 14 days. HSR14 could reach >100% of the control's growth at 200 microg ml(-1) but the SRA reached only up to 60% of the control without metal at 100 microg ml(-1). Though the concentration of Pb was double that of Hg, HSR4 could grow and respire better than the control, the growth being stimulated by 160% and respiration by 170% in the presence of 500 microg ml(-1) of Pb(NO3)2. It is probable that some hypersaline SRB are more tolerant to heavy metals than the mesohaline counterparts and could be more effectively used for precipitating these metals in bioremediatory measures. Further examination of their responses to varied concentration of metals under different salinities would indicate their range of applicability.

Adaptation, Physiological↗