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The bacterial diversity in sediments from the northern Bering Sea was investigated by culture-independent approaches. Community fingerprint analysis by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) revealed that sediment at two deep stations (DBSE and DBS1, >400 m in depth) harbored a bacterial community distinct from the sediments collected at shallow stations (<150 m in depth) on the continental shelf. Three 16S rRNA gene clone libraries for sediments collected from shallow to deep water stations (NEC5, DBSE and DBS1, respectively) were established. Sediment collected at the deepest station DBS1 showed the highest diversity index value. Sequences fell into 19 major lineages of the domain Bacteria: Alpha-, Beta-, Gamma-, Delta- and Epsilonproteobacteria, Bacteroidetes, Acidobacteria, Actinobacteria, Firmicutes, Planctomycetes, Nitrospirae, Verrucomicrobia, Chloroflexi, Chlorobi, Spirochaetes, Cyanobacteria (or chloroplasts), and candidate divisions OP8, TM6, and WS3. A small fraction of retrieved sequences (1.8%) did not fall into any taxonomic division. Deltaproteobacteria (30%) was the dominant phylum in the three libraries, followed by Gammaproteobacteria (21%) and Acidobacteria (16%). The percentages of cloned sequences with the highest similarity to reported sequences below 97 and 93% were 48.1 and 24.3%, respectively. A large number of phylotypes affiliated with bacteria that play important roles in the carbon, sulfur, and nitrogen cycles suggest an important link of bacteria to the matter cycling in these subarctic sediments. [ABSTRACT FROM AUTHOR]

Oxidation of Pu(III) in 1 bp solution to Pu(IV) was studied using the salt-free oxidant NO. It was proved that the reductants N,N-dimethylhydroxylamine (DMHAN) and monomethyl-hydrazine (MMH) present in 1 bp solution of CIAE-APOR process can be oxidized and removed from the solutions also by NO before the oxidation of Pu(III). The effects of the acidity, the temperature and the amount of NO added on the oxidation of DMHAN and MMH were studied. [ABSTRACT FROM AUTHOR]

Microbial eukaryote diversity was assessed in Arctic Kongsfjorden (Svalbard), by constructing SSU rDNA clone libraries. Samples were collected from different depths at the outer basin in summer (2006), plus an additional one glacial and one sediment sample. The libraries displayed diversity based on 284 full-length sequences. Four main phyla, namely, Alveolates, Stramenopiles, Cercozoans, and Metazoans were often screened in this fjord. Alveolate occupied the highest percentage of taxa in the library of surface sea water, besides the Metazoan-related clones. Moreover, dinoflagellates, diatoms, and pico-Prasinophytes were detected as prevalent phytoplankton through the analysis of libraries. Questions related to the quantity of these phytoplankton and their roles in the microbial food loop arose from an ecological viewpoint. [ABSTRACT FROM AUTHOR]

Phylogenetic diversity of the marine bacterioplankton in Kongsfjorden (Spitsbergen) was investigated by 16S rRNA gene analysis. Community fingerprint analysis by PCR-denaturing gradient gel electrophoresis revealed that there was no apparent difference of bacterioplankton community composition between sampling locations in the fjord. A higher biodiversity was observed in bottom water of station 3 in the central part of the fjord. By 16S rRNA gene clone library analysis, sequences detected both in surface and bottom water of station 3 fell into eight putative divisions, including Proteobacteria (Alpha, Beta, Gamma and Delta), Bacteroidetes, Actinobacteria, Verrucomicrobia and unidentified bacteria, in addition to chloroplasts of algae. Sequences representing Planctomycetes were only observed in bottom water. Compared to the preponderance of clones representing Gammaproteobacteria (36.5%) and Alphaproteobacteria (29.4%) in bottom water, Alphaproteobacteria (43.6%) and algae (27.7%) constituted two dominant fractions in surface water. Cloned sequences showed 82.1–100% similarity to those described sequences. It suggests that, attributing to the influence of ocean currents as well as freshwater input in the summer, the bacterial community in Kongsfjorden may consist of a mixture of cosmopolitan and uniquely endemic phylotypes. [ABSTRACT FROM AUTHOR]

A total of 338 aerobic heterotrophic bacterial strains were isolated from Arctic sea ice, Canada Basin (77°30′N–80°12′N). The capability of the isolates to produce protease, lipase, amylase, chitinase, β-galactosidase, cellulase and/or agarase was investigated. Isolates that were able to degrade tributyrin, skim milk, starch, lactose and chitin accounted for 71.6, 65.7, 38.5, 31.6 and 16.9% of sea ice strains, respectively. Lipase producers and/or protease producers were phylogenetically widespread among the isolated strains. Starch and/or lactose hydrolytic strains were mainly distributed among Colwellia, Marinomonas, Pseudoalteromonas, Pseudomonas and Shewanella isolates. Pseudoalteromonas tetraodonis, Pseudoalteromonas elyakovii, Bacillus firmus and Janibacter melonis isolates all have the ability to degrade chitin. Only some strains belonging to Pseudoalteromonas genus scored positive for agarase (6) and cellulose (9). The temperature dependences for lipase activities were determined for five psychrophilic and six psychrotolerant bacteria. At low temperatures, the psychrophilic bacterial lipase activity was not significantly higher than psychrotolerant bacterial lipase, though all lipases showed remarkably high activity with 10–36% residual activity at 0°C. [ABSTRACT FROM AUTHOR]

In order to assess bacterial diversity within four surface sediment samples (0–5 cm) collected from the Pacific Arctic Ocean, 16S ribosomal DNA clone library analysis was performed. Near full length 16S rDNA sequences were obtained for 463 clones from four libraries and 13 distinct major lineages of Bacteria were identified (α, β, γ, δ and ε-Proteobacteria, Acidobacteria, Bacteroidetes, Chloroflexi, Actinobacteria, Firmicutes, Planctomycetes, Spirochetes, and Verrucomicrobia). α, γ, and δ-Proteobacteria, Acidobacteria, Bacteroidetes, Actinobacteria were common phylogenetic groups from all the sediments. The γ-Proteobacteria were the dominant bacterial lineage, representing near or over 50% of the clones. Over 35% of γ-Proteobacteria clones of four clone library were closely related to cultured bacterial isolates with similarity values ranging from 94 to 100%. The community composition was different among sampling sites, which potentially was related to geochemical differences. [ABSTRACT FROM AUTHOR]

We studied the microbial diversity in the sediment from the Kongsfjorden, Svalbard, Arctic, in the summer of 2005 based on the analysis of 16S rRNA and 18S rRNA gene clone libraries. The sequences of the cloned 16S rRNA and 18S rRNA gene inserts were used to determine the species identity or closest relatives by comparison with sequences of known species. Compared to the other samples acquired in Arctic and Antarctic, which are different from that of ours, the microbial diversity in our sediment is much higher. The bacterial sequences were grouped into 11 major lineages of the domain Bacteria: Proteobacteria (include α-, β-, γ-, δ-, and ε-Proteobacteria); Bacteroidetes; Fusobacteria; Firmicutes; Chloroflexi; Chlamydiae; Acidobacteria; Actinobacteria; Planctomycetes; Verrucomicrobiae and Lentisphaerae. Crenarchaeota were dominant in the archaeal clones containing inserts. In addition, six groups from eukaryotes including Cercozoa, Fungi, Telonema, Stramenopiles, Alveolata, and Metazoa were identified. Remarkably, the novel group Lentisphaerae was reported in Arctic sediment at the first time. Our study suggested that Arctic sediment as a unique habitat may contain substantial microbial diversity and novel species will be discovered. [ABSTRACT FROM AUTHOR]

The gene encoding purine nucleoside phosphorylase (PNP) from the cold-adapted marine bacterium Pseudoalteromonas sp. Bsi590 was identified, cloned and expressed in Escherichia coli. The gene encodes a polypeptide of 233 amino acids with a calculated molecular weight of 25,018 Da. Pseudoalteromonas sp. Bsi590 PNP ( PiPNP) shares 60% amino sequence identity and conservation of amino acid residues involved in catalysis with mesophilic Escherichia coli deoD-encoded purine nucleoside phosphorylase ( EcPNP). N-terminal his-tagged PiPNP and EcPNP were purified to apparent homogeneity using Ni2+-chelating column. Compared with EcPNP, PiPNP possessed a lower temperature optimum and thermal stability. As for PNP enzymes in general, PiPNP and EcPNP displayed complicated kinetic properties; PiPNP possessed higher K m and catalytic efficiency ( k cat/ K m ) compared to EcPNP at 37°C. Substrate specificity results showed PiPNP catalyzed the phosphorolytic cleavage of 6-oxopurine and 6-aminopurine nucleosides (or 2-deoxynucleosides), and to a lesser extent purine arabinosides. PiPNP showed a better activity with inosine while no activity toward pyrimidine nucleosides. The protein conformation was analyzed by temperature perturbation difference spectrum. Results showed that PiPNP had lower conformation transition point temperature than EcPNP; phosphate buffer and KCl had significant influence on PiPNP protein conformation stability and thermostability. [ABSTRACT FROM AUTHOR]

To choose a suitable restriction endonuclease for quick assessment of bacterial diversity in polar environments by ARDRA, we investigated the effect of restriction enzymes on ARDRA patterns of cultivable marine planktonic bacteria isolated from polar region. Thirty-three isolates were analyzed by ARDRA using five enzymes ( HinfI, HaeIII, AluI, and the mix AfaI/ MspI), respectively, resulting in different groups, each group corresponding to a particular genotype. A comparison of the ARDRA patterns was carried out, and phylogenetic position of all thirty-three bacteria was obtained by 16S rDNA sequencing. Consistent with phylogenetic analysis, ARDRA pattern comparison revealed that AluI, being sensitive and reliable enough to generate species-specific patterns, was a suitable restriction enzyme used for evaluating bacterial diversity, suggesting a combination of ARDRA with AluI and 16S rDNA sequencing can provide a simple, fast and reliable means for bacterial identification and diversity assessment in polar environments. [ABSTRACT FROM AUTHOR]