Extracellular enzymatic activities of cold-adapted bacteria from polar oceans and effect of temperature and salinity on cell growth

The potential of 324 bacteria isolated from different habitats in polar oceans to produce a variety of extracellular enzymatic activities at low temperature was investigated. By plate assay, lipase, protease, amylase, gelatinase, agarase, chitinase or cellulase were detected. Lipases were generally present by bacteria living in polar oceans. Protease-producing bacteria held the second highest proportion in culturable isolates. Strains producing amylase kept a relative stable proportion of around 30% in different polar marine habitats. All 50 Arctic sea-ice bacteria producing proteases were cold-adapted strains, however, only 20% were psychrophilic. 98% of them could grow at 3% NaCl, and 56% could grow without NaCl. On the other hand, 98% of these sea-ice bacteria produced extracellular proteases with optimum temperature at or higher than 35℃, well above the upper temperature limit of cell growth. Extracellular enzymes including amylase, agarase, cellulase and lipase released by bacteria from seawater or sediment in polar oceans, most expressed maximum activities between 25 and 35℃. Among extracellular enzymes released by bacterial strain BSw20308, protease expressed maximum activity at 40℃, higher than 35℃ of polysaccharide hydrolases and 25℃ of lipase.

Phylogenetic diversity and phenotypic characterization of cultivable bacterioplankton isolated from polar oceans

A set of 27 marine planktonic bacteria isolated from the polar regions was characterized by 16S rDNA sequencing and physiological and biochemical testing. More than half of these bacteria were positive for caseinase, gelatinase and β-glucosidase, and could utilize glucose, maltose or malic acid as carbon source for cell growth. Twelve isolates expressed nitrate reduction activities. Except for one antarctic isolate BSw10175 belonging to Actinobacteria phylum, these isolates were classified as γ-Proteobacteria, suggesting that γ-Proteobacteria dominated in cultivable marine bacterioplankton at both poles. Genus Pseudoalteromonas was the predominant group in the Chukchi Sea and the Bering Sea, and genus Shewanella dominated in cultivable bacterioplankton in the Prydz Bay. With sequence similarities above 97%, genus Psychrobacter was found at both poles. These 27 isolates were psychrotolerant, and significant 16S rDNA sequence similarities were found not only between arctic and antarctic marine bacteria (>99%), but also between polar marine bacteria and bacteria from other aquatic environments (≥98.8%), including temperate ocean, deep sea, pond and lake, suggesting that in the polar oceans less temperature-sensitive bacteria may be cosmopolitan and have a bipolar, even global, distribution at the species level.

Sensitivity of the Arctic sea ice concentration forecasts to different atmospheric forcing: a case study

A regional Arctic configuration of the Massachusetts Institute of Technology general circulation model(MITgcm) is used as the coupled ice-ocean model for forecasting sea ice conditions in the Arctic Ocean at the National Marine Environmental Forecasting Center of China(NMEFC), and the numerical weather prediction from the National Center for Environmental Prediction Global Forecast System(NCEP GFS) is used as the atmospheric forcing. To improve the sea ice forecasting, a recently developed Polar Weather Research and Forecasting model(Polar WRF) model prediction is also tested as the atmospheric forcing. Their forecasting performances are evaluated with two different satellite-derived sea ice concentration products as initializations:(1) the Special Sensor Microwave Imager/Sounder(SSMIS) and(2) the Advanced Microwave Scanning Radiometer for EOS(AMSR-E). Three synoptic cases, which represent the typical atmospheric circulations over the Arctic Ocean in summer 2010, are selected to carry out the Arctic sea ice numerical forecasting experiments. The evaluations suggest that the forecasts of sea ice concentrations using the Polar WRF atmospheric forcing show some improvements as compared with that of the NCEP GFS.

Flow cytometry investigation of picoplankton across latitudes and along the circum Antarctic Ocean

Using a flow cytometer (FCM) onboard the R/V Xuelong during the 24th Chinese Antarctic cruise, picoplankton community structure and biomass in the surface water were examined along the latitude and around the Antarctic Ocean. Salinity and temperature were automatically recorded and total Chl a was determined. Along the cruise, the abundance of Synechococcus, Prochlorococcus, pico-eukaryotes and heterotrophic bacteria ranged in 0.001-1.855 × 10 8 ind./L, 0.000-2.778 × 10 8 ind./L, 0.002-1.060 × 10 8 ind./L and 0.132-27.073 × 10 8 ind./L, respectively. Major oceanic distribution of Synechococcus and Prochlorococcus appeared between latitudes 30 N and 30 S. Prochlorococcus was mainly influenced by water temperature, water mass combination and freshwater inflow. Meanwhile, Synechococcus distribution was significantly associated with landing freshwater inflow. Pico-eukaryotes and heterotrophic bacteria were distributed all over the oceans, but with a relatively low abundance in the high latitudes of the Antarctic Ocean. Principal Component Analysis showed that at same latitude of Atlantic Ocean and Indian Ocean, picoplankton distribution and constitution were totally different, geographical location and different water masses combination would be main reasons.

Phylogenetic diversity of dimethylsulfoniopropionate-dependent demethylase gene dmdA in distantly related bacteria isolated from Arctic and Antarctic marine environments

Dimethylsulfoniopropionate(DMSP) is mainly produced by marine phytoplankton as an osmolyte, antioxidant,predator deterrent, or cryoprotectant. DMSP is also an important carbon and sulfur source for marine bacteria.Bacteria may metabolize DMSP via the demethylation pathway involving the DMSP demethylase gene(dmdA) or the cleavage pathway involving several different DMSP lyase genes. Most DMSP released into seawater is degraded by bacteria via demethylation. To test a hypothesis that the high gene frequency of dmdA among major marine taxa results in part from horizontal gene transfer(HGT) events, a total of thirty-one bacterial strains were isolated from Arctic Kongsfjorden seawater in this study. Analysis of 16S rRNA gene sequences showed that,except for strains BSw22118, BSw22131 and BSw22132 belonging to the genera Colwellia, Pseudomonas and Glaciecola, respectively, all bacteria fell into the genus Pseudoalteromonas. DmdA genes were detected in five distantly related bacterial strains, including four Arctic strains(Pseudoalteromonas sp. BSw22112, Colwellia sp.BSw22118, Pseudomonas sp. BSw22131 and Glaciecola sp. BSw22132) and one Antarctic strain(Roseicitreum antarcticum ZS2–28). Their dmdA genes showed significant similarities(97.7%–98.3%) to that of Ruegeria pomeroyi DSS–3, which was originally isolated from temperate coastal seawater. In addition, the sequence of the gene transfer agent(GTA) capsid protein gene(g5) detected in Antarctic strain ZS2–28 exhibited a genetically closely related to that of Ruegeria pomeroyi DSS–3. Among the five tested strains, only Pseudomonas sp. BSw22131 could grow using DMSP as the sole carbon source. The results of this study support the hypothesis of HGT for dmdA among taxonomically heterogeneous bacterioplankton, and suggest a wide distribution of functional gene(i.e., dmdA) in global marine environments.

Dome Argus: Ideal site for deep ice drilling

Located on the centre of ice drainage range, the highest Dome Argus (Dome A) of East Antarctic Ice Sheet (EAIS), could be represented as an ideal site for deep ice cores drilling containing oldest paleo-climate records. To select a suitable drilling site for deep ice core, it needs gather all information pertaining to the local meteorology, ice sheet landforms, ice thickness, subglacial topography of bed rocks, ice velocity, internal structures of ice sheet, etc. Based on the International Partnerships in Ice Core Sciences (IPICS), we present recent achievement of glaciological research and its perspective at Dome A in this paper. We system-atically discussed the merits and possible ventures of potential drilling sites around Dome A. Among all the candidates, we find that the Chinese Antarctic Kunlun Station is the best site for carrying out the first deep ice core drilling campaign. We emphasize and assess further the possibility to obtain a replicate core for studying dynamics and evolution of climate change.

Diversity of the aerobic anoxygenic phototrophy gene pufM in Arctic and Antarctic coastal seawaters

Aerobic anoxygenic phototrophic(AAP) bacteria serve important functions in marine carbon and energy cycling because of their capability to utilize dissolved organic substrates and harvest light energy.AAP bacteria are widely distributed in marine environments,and their diversity has been examined in marine habitats.However,information about AAP bacteria at high latitudes remains insufficient to date.Therefore,this study determined the summer AAP bacterial diversity in Arctic Kongsfjorden and in the Antarctic coastal seawater of King George Island on the basis of puf M,a gene that encodes a pigment-binding protein subunit of the reaction center complex.Four puf M clone libraries were constructed,and 674 positive clones were obtained from four investigated stations(two in Kongsfjorden and two in the Antarctic Maxwell Bay).Arctic clones were clustered within the Alphaproteobacteria,whereas Antarctic clones were classified into the Alphaproteobacteria and Betaproteobacteria classes.Rhodobacteraceae-like puf M genes dominated in all samples.In addition,sequences closely related to puf M encoded on a plasmid in Sulfitobacter guttiformis were predominant in both Arctic and Antarctic samples.This result indicates the transpolar or even global distribution of puf M genes in marine environments.Meanwhile,differences between the Arctic and Antarctic sequences may prove polar endemism.These results indicate the important role of Rhodobacteraceae as AAP bacteria in bipolar coastal waters.

Reduction of hydrocarbon contamination on viability of Acartia pacifica benthic resting eggs

The potential effect of hydrocarbon contamination on the hatching success of benthic resting eggs of Acartia pacifica in Xiamen Bay was investigated experimentally. The number of nauplii emerging from the sediment samples decreased with increasing Fuel Oil #0 concentration. The estimated rate of mortality increased markedly with the increase of Fuel Oil #0 concentration. Successive fuel Oil #0 concentrations from 50 mg/kg to 5000 mg/kg reduced the number of hatched nauplii by 3.8%-100%. The mortality of A. pacifica resting eggs due to Fuel Oil #0 contamination did not significantly increase as time progressed at each concentration level. The LC50 values of resting eggs, changing from 237.12 to 279.59 mg/kg, remained at an almost stable level in two months. The number of A. pacifica nauplii that hatched from the sediment at 10°C was higher than those from the sediment at 30°C, which indicates that the toxicity of Fuel Oil #0 on A. pacifica resting eggs increases with increasing temperature.

Phylogenetic diversity of planktonic bacteria in the Chukchi Borderland region in summer

Planktonic bacteria are abundant in the Chukchi Borderland region. However, little is known about their diversity and the roles of various bacteria in the ocean. Seawater samples were collected from two stations K2S and K4S where sea ice was melting obviously. The analysis of water samples with fluorescence in situ hybridization (FISH) showed that DMSP-degrading bacteria accounted for 13% of the total bacteria at the station K2S. No aerobic anoxygenic phototrophic (AAP) bacteria were detected in both samples. The bacterial communities were characterized by two 16S rRNA gene clone libraries. Sequences fell into four major lineages of the domain Bacteria, including Proteobacteria (Alpha, Beta and Gamma subclasses), Bacteroidetes, Actinobacteria and Firmicutes. No significant difference was found between the two clone libraries. SAR11 and Rhodobacteraceae clades of Alphaproteobacteria and Pseudoalteromonas of Gammapro-teobacteria constituted three dominant fractions in the clone libraries. A total of 191 heterotrophic bacterial strains were isolated and 76% showed extracellular proteolytic activity. Phylogenetic analysis reveals that the isolates fell into Gammaproteobacteria, Bacteroidetes, Actinobacteria and Firmicutes. The most common genus in both the bacterial isolates and protease-producing bacteria was Pseudoalteromonas. UniFrac data showed suggestive differences in bacterial communities between the Chukchi Borderland and the northern Bering Sea.

Preliminary investigations on Arctic microalgae by jointapplication of fluorescent instruments

In vivo fluorescence has a wide application in analyzing microalgae, including assessing phytoplankton biomass, ratesof primary production and physiological status. This study describes a preliminary investigation on the joint application of the threekinds of fluorescence analysis in the physiological study of microalgae. Flow cytometry and fluorescence spectrometry were usedto obtain the in vivo static fluorescence information of pigments, and a Pulsed-Amplitude-Modulation chlorophyll fluorometer wasused to detect the dynamic fluorescence of chlorophyll. The validity of the joint application was proved by analyzing two labora-tory cultured Arctic microalgae, Pseudo-nitzschia delicatissima (Bacillariophyceae) and Thalassiosira sp. The higher value ofminimum fluorescence yield in dark-adapted state (Fo), actual photochemical efficiency of PSII (ΦPSII), and electron transportrate (ETR) exhibited positive results in a higher cell abundance and chlorophyll a content of P. delicatissima; whereas higherβ-carotene content of Thalassiosira sp. played an important role in the protection of photosynthesis.

Population dynamics of Acartia pacifica (Copepoda:Calanoida):the importance of benthic-pelagic coupling

The seasonal occurrence of Acartia pacifica (Copepoda: Calanoida) and their resting eggs in the sediment of Xiamen Bay were documented between October 2002 and September 2003. The number of viable eggs in the sediment increased from January to May with the increase in the number of planktonic females. When the population of A. pacifica disappeared from the water column, the number of eggs in the sediment began to decrease and reached a low value due to lack of input. The peak of nauplii abundance occurred when the hatching potential of eggs from the sediment was high under the natural environment from February to June. The hatching of resting eggs of A. pacifica was essentially temperature-dependent after suspension, while photoperiod regimes had no significant effect on the hatching. The mean density of subitaneous eggs was 1.122 0 g/cm 3 with a standard deviation (SD) of 0.000 2 g/cm 3 . The mean density of diapause eggs was 1.151 2 g/cm 3 with a SD of 0.000 1 g/cm 3 . The sinking rates of subitaneous eggs ranged from 19.55 to 26.17 m/d, while those of diapause eggs ranged from 30.29 to 31.28 m/d. The comparison of the egg deposition time and egg hatching time suggested that in most cases virtually all subitaneous eggs of A. pacifica would settle to the bottom before their hatching even though the eggs have high potential to hatch. The evidence was provided that the seasonal dynamics of A. pacifica is accompanied by benthic-pelagic coupling.

Simulation of arctic surface radiation and energy budget during the summertime using the single-column model

The surface heat budget of the Arctic Ocean (SHEBA) project has shown that the study of the surface heat budget characteristics is crucial to understanding the interface process and environmental change in the polar region. An arctic single-column model (ARCSCM) of Colorado University is used to simulate the arctic surface radiation and energy budget during the summertime. The simulation results are analyzed and compared with the SHEBA measurements. Sensitivity analyses are performed to test microphysical and radiative parameterizations in this model. The results show that the ARCSCM model is able to simulate the surface radiation and energy budget in the arctic during the summertime,and the different parameterizations have a significant influence on the results. The combination of cloud microphysics and RRTM parameterizations can fairly derive the surface solar shortwave radiation and downwelling longwave radiation flux. But this cloud microphysics parameterization scheme deviates notably from the simulation of surface sensible and latent heat flux. Further improvement for the parameterization scheme applied to the Arctic Regions is necessary.

Relationships between two Pseudoalteromonas strains isolated from the Canada Basin and the Southern Ocean using a polyphasic approach

The evolutionary relationships and taxonomic position of two marine planktonic bacterial strains BSw20211 and BSw10014,isolated from the Canada Basin and from the Southern Ocean,respectively,were determined using a polyphasic taxonomic approach.There was a close phylogenetic relationship between the two strains and most phenotypic properties were shared.Nonetheless,they were found to belong to different species of the genus Pseudoalteromonas on the basis of genotypic analyses.Findings were consistent with the suggestion that gyrB gene sequence comparison and DNA-DNA relatedness might better define phylogenetic relationships of bacteria at the species level.However,a cut-off value of 90% gyrB gene sequence similarity was not reliable for the differentiation of species within the genus Pseudoalteromonas.

Abundance,biomass and composition of spring ice algal and phytoplankton communities of the Laptev Sea(Arctic)

Abundance, biomass and composition of the ice algal and phytoplankton communities were investigated in the southeastern Laptev Sea in spring 1999.Diatoms dominated the algal communities and pennate diatoms dominated the diatom population. 12 dominant algal species occurred within sea ice and underlying water column, including Fragilariopsis oceanica, F. cylindrus, Nitzschia frigida , N. promare, Achnanthes taeniata , Nitzschia neofrigida , Navicula pelagica , N. vanhoef fenii, N. septentrionalis, Melosira arctica , Clindrotheca closterium and Pyramimonas sp. The algal abundance of bottom 10 cm sea ice varied between 14.6 and 1562.2 × 104 cells l-1 with an average of 639.0 × 104cells l-1 , and the algal biomass ranged from 7.89 to 2093.5 μg C l-1 with an average of 886.9 μg C l-1 , which were generally one order of magnitude higher than those of sub-bottom ice and two orders of magnitude higher than those of underlying surface water. The integrated algal abundance and biomass of lowermost 20 cm ice column were averagely 7.7 and 12.2 times as those of upper 20 m water column, respectively, suggesting that the ice algae might play an important role in maintaining the coastal marine ecosystem before the thawing of sea ice. Ice algae influenced the phytoplankton community of the underlying water column.However, the "seeding" of ice algae for phytoplankton bloom was negligible because of the low phytoplankton biomass within the underlying water column.

Isolation of novel psychrophilic bacteria from Arctic sea ice

The phylogenetic diversity of culturable psychrophilic bacteria associ ated with sea ice from the high latitude regions of Canadian Basin and Chukchi S ea,Arctic,was investigated.A total of 34 psychropilic strains were isolated u sing three methods of(Ⅰ)dilution plating(at 4 ℃),(Ⅱ)bath culturing(at-1 ℃)and dilution plating,and(Ⅲ)cold shock(-20 ℃ for 24 h),bath culturin g and dilution plating under aerobic conditions.Sea-ice samples were exposed to-20 ℃ for 24 h that might reduce the number of common microorganisms and encou rag e outgrowth of psychrophilic strains.This process might be able to be introduce d to isolation psychrophilic bacteria from other environmental samples in future study.16S rDNA nearly full-length sequence analysis revealed that psychrophil i c strains felled in two phylogenetic divisions,γ-proteobacteria(in the gen era Colwellia、Marinobacter、Shewanella、Glaciecola、Marinomonas and Pseudoalt eromon as) and Cytophaga-Flexibacter-Bacteroides(Flavobacterium and Psychrof lexus).Fi fteen of bacterial isolates quite likely represented novel species(16S rDNA seq uence similarity below 98%).One of strains(BSi20002)from Canadian Basin showe d 100% sequence similarity to that of Marinobacter sp.ANT8277 isolated from the Antarctic Weddell sea ice,suggesting bacteria may have a bipolar distribution at the species level.

Determination of trace level of perchlorate in Antarctic snow and ice by ion chromatography coupled with tandem mass spectrometry using an automated sample on-line preconcentration method

A new ion chromatography coupled with tandem mass spectrometry(IC-ESI-MS/MS) method,with automated sampling and on-line preconcentration,has been developed for the determination of perchlorate in Antarctic snow and ice at low part-per-trillion(ng/L) levels.To the best of our knowledge, this is the first time that an analytical method is used for the determination of perchlorate in Antarctic snow and ice.The IC-ESI-MS/MS instrumentation consisted of an ICS2000 ion chromatography(IC) system coupled to an API3200 electrospray tandem mass spectrometer(ESI-MS/MS).On-line preconcentration was realized through a six-port injector valve,a TAC-ULP1 concentrator column and an AS auto-sampler.Multiple reaction monitoring(MRM) mode was used to quantify the perchlorate anion.The transition of ^(35)Cl^(16)O_4^-(m/z 98.9) into ^(35)Cl^(16)O_3^-(m/z 82.9) was monitored for quantifying the main analyte,and the transition of ^(37)Cl^(16)O_4^-(m/z 100.9) into ^(37)Cl^(16)O_3^-(m/z 84.9) was monitored for examining a proper isotopic abundance ratio of ^(35)Cl to ^(37)Cl,which was used as a confirmation tool.The limit of detection(LOD) and limit of quantitation(LOQ) for the method was 0.2 ng/ L and 0.5 ng/L,respectively.And this new method exhibited acceptable accuracy and precision for samples at ng/L levels.All the tested snow and ice samples were found to contain measurable amount of perchlorate,ranging from 10 ng/L to 340 ng/L.

Denaturing gradient gel electrophoresis fingerprinting of soil bacteria in the vicinity of the Chinese Great Wall Station, King George Island, Antarctica

Bacterial diversity was investigated in soil samples collected from 13 sites around the Great Wall Station, Fildes Peninsula, King George Island, Antarctica, using denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes. The classes α-, β-, and γ-Proteobacteria, as well as the phylum Actinobacteria, were found to be the dominant bacteria in the soils around the Great Wall Station. Although the selected samples were not contaminated by oil, a relationship between soil parameters, microbial biodiversity, and human impact was still seen. Sample sites in human impacted areas showed lower bacterial biodiversity (average H′= 2.65) when compared to nonimpacted sites (average H′= 3.05). There was no statistically significant correlation between soil bacterial diversity and total organic carbon (TOC), total nitrogen, or total phosphorus contents of the soil. Canonical correlation analysis showed that TOC content was the most important factor determining bacterial community profiles among the measured soil parameters. In conclusion, microbial biodiversity and community characteristics within relatively small scales (1.5 km) were determined as a function of local environment parameters and anthropogenic impact.