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The Sansha Yongle Blue Hole is the world’s deepest (301 m) underwater cave and has a sharp redox gradient, with oligotrophic, anoxic, and sulfidic bottom seawater. In order to discover the microbial communities and their special biogeochemical pathways in the blue hole, we analyzed the 16S ribosomal RNA amplicons and metagenomes of microbials from seawater depths with prominent physical, chemical, and biological features. Redundancy analysis showed that dissolved oxygen was the most important factor affecting the microbial assemblages of the blue hole and surrounding open sea waters, and significantly explained 44.7% of the total variation, followed by silicate, temperature, sulfide, ammonium, methane, nitrous oxide, nitrate, dissolved organic carbon, salinity, particulate organic carbon, and chlorophyll a. We identified a bloom of Alteromonas (34.9%) at the primary nitrite maximum occurring in close proximity to the chlorophyll a peak in the blue hole. Genomic potential for nitrate reduction of Alteromonas might contribute to this maximum under oxygen decrease. Genes that would allow for aerobic ammonium oxidation, complete denitrification, and sulfur-oxidization were enriched at nitrate/nitrite-sulfide transition zone (90 and 100 m) of the blue hole, but not anammox pathways. Moreover, γ-Proteobacterial clade SUP05, ε-Proteobacterial genera Sulfurimonas and Arcobacter, and Chlorobi harbored genes for sulfur-driven denitrification process that mediated nitrogen loss and sulfide removal. In the anoxic bottom seawater (100-300 m), high levels of sulfate reducers and dissimilatory sulfite reductase gene (dsrA) potentially created a sulfidic zone of ~200 m thickness. Our findings suggest that in the oligotrophic Sansha Yongle Blue Hole, O2 deficiency promotes nitrogen- and sulfur-cycling processes mediated by metabolically versatile microbials.

Abstract: Chitosan is derived from chitin polysaccharide, the main component of crustacean shells. Chitosan is a biocompatible, nontoxic, and biodegradable polymer soluble in acidic solutions. It is widely used in the medical and pharmaceutical fields. Antimicrobial activities of chitosan against different bacterial, fungal, and viral pathogens have been considered one of its attractive properties, making chitosan valuable for biological applications, including textile, food, tissue engineering, agriculture, and environmental protection. Additionally, chitosan has beneficial effects on livestock, poultry, fish, and crustaceans, which can enhance immunity, improve feed conversion, and promote growth. However, the water solubility of chitosan influences antimicrobial capabilities, limiting its application. In the present work, we reviewed the preparation, factors affecting antimicrobial activity, morphological structure, antimicrobial mechanism, and application of chitosan derivatives, and the problems and prospects were pointed out. Collectively, this review provided an update on the application of chitosan derivatives and their potential for further advanced applications in the antimicrobial field.

Mucosal tissue constitutes the largest interface between the body and the external environment, regulating the entry of pathogens, particles, and molecules. Mucosal immunization is the most effective way to trigger a protective mucosal immune response. However, the majority of the currently licensed vaccines are recommended to be administered by intramuscular injection, which has obvious shortcomings, such as high production costs, low patient compliance, and lack of mucosal immune response. Strategies for eliciting mucosal and systemic immune responses are being developed, including appropriate vaccine adjuvant, delivery system, and bacterial or viral vectors. Biodegradable mucoadhesive nanoparticles (NPs) are the most promising candidate for vaccine delivery systems due to their inherent immune adjuvant property and the ability to protect the antigen from degradation, sustain the release of loaded antigen, and increase the residence time of antigen at the administration site. The current review outlined the complex structure of mucosa, the mechanism of interaction between NPs and mucosa, factors affecting the mucoadhesion of NPs, and the application of the delivery system based on mucoadhesive NPs in the field of vaccines. Moreover, this review demonstrated that the biodegradable and mucoadhesive NP-based delivery system has the potential for mucosal administration of vaccines.

Colwellia sp. Arc7-635, a psychrophilic denitrifying bacterium isolated from Arctic seawater, uses NO3− or NH4+ as the sole nitrogen source to grow at low temperatures. In this article, we describe the complete genome of Colwellia sp. Arc7-635. The genome has one circular chromosome of 4,741,350 bp (38.41 mol% G+C content), consisting of 3841 coding genes, 91 tRNA genes, as well as seven rRNA operons of 16S-23S-5S rRNA, and one operon of 16S-23S-5S-5S rRNA. According to the genomic annotation results, strain Colwellia sp. Arc7-635 encodes a complete denitrifying pathway consisting of genes affiliated with nitrate reductase, nitrite reductase, nitric oxide reductase, and nitrous oxide reductase. Genes affiliated with nitrate reduction to ammonia including nitrate reductases (NapA and NapB) and nitrite reductases (NirA, NirB, and NirD) were also identified. The whole genome sequences of Arc7-635 provide information that is useful for further clarifying nitrogen metabolisms and facilitate its potential applications in the bioremediation of nitrogen pollutions.

Silver nanoparticles (AgNPs) have attracted wide interest due to its broad range of applications. This study aims to describe the biosynthesis of AgNPs using an Arctic anti-oxidative bacterium Arc7-R13 and to study its characteristics and antibacterial activity. The biosynthesis of AgNPs was verified using UV-Vis spectrum with the maximum absorption at 416 nm. The morphology of the silver nanoparticles was characterized by TEM and its characterization were investigated by EDX and FTIR. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain Arc7-R13 was affiliated with genus Paracoccus. TEM analysis revealed that the AgNPs synthesized by strain Arc7-R13 were spherical and ellipsoidal in shape with size ranging from 2 to 25 nm. The optimal concentration of AgNO3 and temperature for the biosynthesis were 4 mmol/L and 37 °C, respectively. EDX analysis verified the presence of the element silver in the biosynthesized AgNPs. FTIR analysis revealed that the specific functional groups, OH, CH3 and C≡N, might be responsible for reduction and stabilization of AgNPs. Antimicrobial test showed that the AgNPs had strong antimicrobial activity against all kinds of strains investigated, including Gram-positive Bacillus subtilis, Staphylococcus aureus and Gram-negative Pseudomonas aeruginosa, Escherichia coli.

To expand investigations and insights into the phylogenetic diversity of bacteria inhibiting seafloor biosphere, six Arctic Ocean sediments neighboring the Bering Strait were sampled and their bacterial diversities were investigated by pyrosequencing of 16S rRNA genes. A total of 157,454 trimed sequences were obtained, resulting in 9413 OTUs at the 97% sequence identity (OTU3%). This pyrosequencing allowed detection of higher than 85% of richness estimator Chao1 and Ace at the OTU3% level. Higher coverage (≥0.97) and much less of rare types (singletons, only accounting for 24.5% of all OTU3%) indicated that this pyrosequencing recovered most of bacteria inhabiting these biospheres. At the phylum level, the high relative sequence abundance (42.0% to 63.3%) showed that Proteobacteria was the dominant member at all these sampling sites. At the class level, Deltaproteobacteria, Gammaproteobacteria, and Flavobacteriia composed the majority of bacterial communities, and the relative abundance of Cyanobacteria and Bacilli varied significantly among the six samples. At the genus level, abundant OTUs related with sulfate reduction, including Desulfobulbus and Desulforhopalus, were identified. Shared and unique OTUs analysis revealed that, at the OTU3% level, 508 OTUs were shared by all the six samples, and the number of unique OTUs ranged from 98 (R02) to 195 (NB04). Principal coordinates analysis PCoA analysis revealed that samples C04 and NB04 had the similar communities and were distinct from the others. Canonical correspondence analysis (CCA) revealed that temperature was the most significant factors that correlated with the bacterial community composition. The differences in bacterial compositions and diversities indicate that the similar sediment habitats contain a large variation in microbial biodiversity.

In this study, an endolytic alginate lyase, named Al163, was identified, cloned, and characterized from the Antarctic bacterium Pseudoalteromonas sp. NJ-21. Comparative sequence analysis showed that the predicted amino acid sequence encoded by al163 belongs to the polysaccharide lyase 6 (PL-6) family and has a molecular mass of about 80 kDa. Recombinant enzyme was purified by Ni-Sepharose affinity chromatography. Recombinant Al163 exhibited maximum activity (258 U/mg) at pH 7.0 and 40℃, and thermal stability assays showed retention of almost 90% activity after incubation at 30℃ for 30 min. Al163 activity was stimulated by Cd2+, Ca2+, Fe 3+, and Mn2+, but inhibited by Cu2+, Si2+, Fe2+, and Ni2+. Thin-layer chromatographic analysis indicated that Al163 degraded sodium alginate, polyM, and polyG, generating disaccharides and trisaccharides as the final products. Only a few bacterial strains that produce a bifunctional alginate lyase have been reported. Our results indicate that recombinant Al163 exhibits broad substrate specificity and its products exhibit low degrees of polymerization. Both properties imply high potential for use of the enzyme in several industrial fields, including cosmetics and pharmaceuticals, based on the high demand for biologically active oligosaccharides.

The general features and genome characteristics of the denitrifying bacterium Marinobacter sp. Arc7-DN-1, isolated from Arctic Ocean sediment, are described. Marinobacter sp. Arc7-DN-1 uses NO3− or NH4+ as the sole nitrogen source to grow at low temperatures. The strain can grow at a wide range of temperatures (0–30 °C) and NaCl concentration (15–90‰). The genome has one circular chromosome of 4,300,456 bp (57.64 mol% G + C content), consisting of 4012 coding genes, including 50 tRNAs and three rRNA operons as 16S-23S-5S rRNA. On the basis of the KEGG analysis, strain Arc7-DN-1 encodes 43 proteins related to nitrogen metabolism, including a complete denitrifying pathway and an assimilatory nitrate reduction pathway.

Colwellia sp. Arc7-D, a psychrophilic H2O2-resisitant bacterium, was isolated from Arctic Ocean sediment. Here we describe the complete genome of Colwellia sp. Arc7-D. The genome has one circular chromosome of 4,305,442 bp (37.67 mol% G + C content), consisting of 3526 coding genes, 77 tRNA genes, as well as five rRNA operons as 16S–23S-5S rRNA and one rRNA operon as 16S-23S-5S-5S. According to KEGG analysis, strain Arc7-D encodes 23 genes related with antioxidant activity including superoxide dismutase, glutathione peroxidase, glutathione reductase and catalase. However, many additional genes affiliated with anti-oxidative stress were also identified, such as aconitase, thioredoxin and ascorbic acid.

This study was to investigate bacterial and archaeal community structure of pan-Arctic Ocean sediments by pyrosequencing. In total, investigation of three marine sediments revealed 15 002 bacterial and 4 362 archaeal operational taxonomic units (OTUs) at the 97% similarity level. Analysis of community structure indicated that these three samples had high bacterial and archaeal diversity. The most relatively abundant bacterial group in Samples CC1 and R05 was Proteobacteria, while Firmicutes was dominant in Sample BL03. Thaumarchaeota was the most relatively abundant archaeal phylum in Samples CC1 and R05, and the relative abundance of Thaumarchaeota was almost as high as that of Euryarchaeota in Sample BL03. These two phyla accounted for nearly 100% of the archaeal OTUs. δ-Proteobacteria and γ-Proteobacteria were the two most relatively abundant classes at Proteobacterial class level, and their relative abundance was more than 60% in Samples CC1 and R05. There were also differences in the top 10 relatively abundant bacterial and archaeal OTUs among the three samples at the 97% similarity, and only 12 core bacterial OTUs were detected. Overall, this study indicated that there were distinct microbial communities and many unique OTUs in these three samples.

The key functional genes involved in temperature adaption of the Antarctic psychrotrophic bacterium Psychrobacter sp. G. were identified by transcriptomic sequencing. We analyzed the global transcriptional profile of Psychrobacter sp. G under cold stress (0°C) and heat stress (30°C), with the optimal growth temperature 20°C as the control. There were large alterations of the transcriptome profile, including significant upregulation of 11 and 12 transcripts as well as significant downregulation of 47 and 42 transcripts in the cold and heat stress groups, respectively, compared to the control. The expression of various genes encoding enzymes and transcriptional regulators, including PfpI and TetR family transcriptional regulators under heat stress, as well as the expression of DEAD/DEAH box helicase and the IclR family of transcriptional regulators under cold stress, were upregulated significantly. The expression of several genes, most affiliated with TonB-dependent receptor and siderophore receptor, was downregulated significantly under both heat and cold stress. Many of the genes associated with the metabolism of fatty acid and ABC transporters were regulated differentially under different temperature stress. The results of this survey of transcriptome and temperature stress-relevant genes contribute to our understanding of the stress-resistant mechanism in Antarctic bacteria.

Oxidative stress is one of the major challenges faced by Arctic marine bacteria due to the high oxygen concentration of seawater, low temperatures and UV radiations. Transcriptome sequencing was performed to obtain the key functional genes involved in the adaptation to oxidative stress induced by hydrogen peroxide in the Arctic bacterium Pseudoalteromonas sp. A2. Exposure to 1 mmol/L H2O2 resulted in large alterations of the transcriptome profile, including significant up-regulation of 109 genes and significant down-regulation of 174 genes. COG functional classification revealed that among the significantly regulated genes with known function categories, more genes belonging to posttranslational modification, protein turnover and chaperones were significantly up-regulated, and more genes affiliated with chaperones and amino acid transport and metabolism were significantly down-regulated. It was notable that the expressions of eighteen genes affiliated with flagella and four genes affiliated with heat shock proteins were significantly up-regulated. Meanwhile, the expression of nine genes belonging to cytochrome and cytochrome oxidase, and five genes belonging to TonB-dependent receptor, were significantly down-regulated. Among the eighteen genes with antioxidant activity categorized by GO analysis, the expression of one gene was significantly up-regulated; however, the expressions of two genes were significantly down-regulated. Briefly, RNA-Seq indicated that, except for the classical anti-oxidative genes and stress proteins, genes affiliated with flagella and function unknown played important roles in coping with oxidative stress in Pseudoalteromonas sp. A2. This overall survey of transcriptome and oxidative stress-relevant genes can contribute to understand the adaptive mechanism of Arctic bacteria.

This study aimed to determine the microbial community structure of seawater in (ICE-1) and out (FUBIAO) of the pack ice zone in the Arctic region. Approximate 10 L seawater was filtrated by 0.2 μm Whatman nuclepore filters and the environmental genomic DNA was extracted. We conducted a detailed census of microbial communities by pyrosequencing. Analysis of the microbial community structures indicated that these two samples had high bacterial, archaeal and eukaryotic diversity. Proteobacteria and Bacteroidetes were the two dominant members of the bacterioplankton community in both samples, and their relative abundance were 51.29% and 35.39%, 72.95% and 23.21%, respectively. Euryarchaeota was the most abundant archaeal phylum, and the relative abundance was nearly up to 100% in FUBIAO and 60% in ICE-1. As for the eukaryotes, no_rank_Eukaryota, Arthropoda and no_rank_Metazoa were the most abundant groups in Sample FUBIAO, accounting for 85.29% of the total reads. The relative abundance of the most abundant phylum in Sample ICE-1, no_rank_Eukaryota and no_rank_Metazoa, was up to 90.69% of the total reads. Alphaproteobacteria, Flavobacteria and Gammaproteobacteria were the top three abundant classes in the two samples at the bacterial class level. There were also differences in the top ten abundant bacterial, archaeal and eukaryotic OTUs at the level of 97% similarity between the two samples.

The general features and genome characteristics of the alginolytic Psychroserpens sp. NJDZ02 isolated from macroalgae collected from King George Island, Antarctica, are described. The sequenced genome is composed of a 4,319,537 bp circular chromosome containing 3699 coding genes including 49 tRNAs and 6 rRNA operons as 16S-23S-5S rRNA. The NJDZ02 genome contains several alginate lyases, which correlate with the high capacity of the strain to degrade sodium alginate. Genome sequencing of NJDZ02 revealed gene candidates of potential interest for use in enzymatic industries or applications in the field of polysaccharide degradation.

Paracoccus sp. Arc7-R13, a silver nanoparticles (AgNPs) synthesizing bacterium, was isolated from Arctic Ocean sediment. Here we describe the complete genome of Paracoccus sp. Arc7-R13. The complete genome contains 4,040,012 bp with 66.66 mol% G + C content, including one circular chromosome of 3,231,929 bp (67.45 mol% G + C content), and eight plasmids with length ranging from 24,536 bp to 199,685 bp. The genome contains 3835 protein-coding genes (CDSs), 49 tRNA genes, as well as 3 rRNA operons as 16S-23S-5S rRNA. Based on the gene annotation and Swiss-Prot analysis, a total of 15 genes belonging to 11 kinds, including silver exporting P-type ATPase (SilP), alkaline phosphatase, nitroreductase, thioredoxin reductase, NADPH dehydrogenase and glutathione peroxidase, might be related to the synthesis of AgNPs. Meanwhile, many additional genes associated with synthesis of AgNPs such as protein-disulfide isomerase, c-type cytochrome, glutathione synthase and dehydrogenase reductase were also identified.

Preoperative acute hypervolemic hemodilution (AHHD) is a technique used in anesthesia to reduce the number of blood cells lost during intraoperative bleeding. The aim of the present study was to evaluate the effect of the hypervolemic hemodilution of 6% hydroxyethyl starch 130/0.4 on the EC50 of propofol at two clinical endpoints. A total of 20 patients undergoing AHHD following epidural anesthesia were studied, and 20 patients who did not receive hemodilution were used as a control group. All patients were American Society of Anesthesiologists grade I, aged 20-40 years and undergoing hip arthroplasty surgery. In the AHHD group, 10 ml/kg lactated Ringer's solution was infused over 20 min at the same time as the epidural test dose. The infusion was followed by the infusion of 6% hydroxyethyl starch 130/0.4 over 30 min. Patients in the control group received 10 ml/kg Ringer's solution over 50 min. Propofol was then delivered by a Diprifusor target-controlled infusion. The predicted blood and effect-site propofol concentrations were recorded at loss of consciousness (LOC) and return of consciousness (ROC). Probit analysis was used to estimate the values for predicted blood and effect-site concentrations at the two clinical endpoints. The results showed that the potency of propofol was decreased during AHHD. Compared with the controls, the predicted blood and effect-site concentrations of propofol at LOC were higher in patients of the hemodilution group, resulting in higher EC50 values (P=0.001 and 0.025, respectively). At ROC, the effect-site EC50 was 2.9 µg/ml [95% confidence interval (CI), 2.8-3.0] in hemodilution patients and 2.5 µg/ml (95% CI, 2.2-2.6) in control patients (P=0.001). With AHHD, the LOC time was significantly longer and the propofol dose was higher, while ROC times were comparable. In conclusion, AHHD increases the requirement for propofol at LOC and prolongs LOC time. Patients with AHHD recovered consciousness at higher effect-site concentrations of propofol. Thus, the induction dose of propofol should be increased during AHHD.

Genome-wide characterization of the Pohlia nutans transcriptome is essential for clarifying the role of stress-relevant genes in Antarctic moss adapting to the extreme polar environment. High-throughput Illumina sequencing was used to analyze the gene expression profile of P. nutans after cold treatment. A total of 93,488 unigenes, with an average length of 405 bp, were obtained. Gene annotation showed that 16,781 unigenes had significant similarity to known functional protein-coding genes, most of which were annotated using the GO, KOG and KEGG pathway databases. Global profiling of the differentially expressed genes revealed that 3,796 unigenes were significantly upregulated after cold treatment, while 1,405 unigenes were significantly downregulated. In addition, 816 receptor-like kinases and 1,309 transcription factors were identified from P. nutans. This overall survey of transcripts and stress-relevant genes can contribute to understanding the stress-resistance mechanism of Antarctic moss and will accelerate the practical exploitation of the genetic resources for this organism.

Psychrobacter has been regarded as an important genus for bacterial cold adaptation studies. However, members of this genus are highly varied in terms of both cold adaptability and genome content. To get an understanding of the diversity of members of this genus, five Psychrobacter strains (G, K5, 273-4, PAMC21119 and PRwf-1), with publicly available complete/draft genome, were selected and comprehensive comparative genomics analyses were performed among them. The closest phylogenetic relationship, highest average nucleotide identity (96.78%) and best sequence synteny were identified between strains G and K5. These findings suggest they belong to the same species, despite the long geographic distance between them (Antarctic and Siberia). 4542 gene clusters in total were identified from the five genomes, and of which 1424 were shared by all of them. The number of genes unique to strains G, K5, 273-4, PAMC21119 and PRwf-1 are 183, 188, 300, 637 and 665, respectively. COG assignment revealed their differences in gene content related to stress response. The extensive sequence rearrangements and the large number of genes unique to strain PAMC21119 and PRwf-1 suggest they may have experienced a high level of gene exchanges in the permafrost soil and the surface of fish skin.

An OmpR gene, named OmpR503, was cloned from the Antarctic psychrotrophic bacterium Psychrobacter sp. G according to its genomic draft. The deduced amino acid sequences of OmpR503 were highly conserved with other known protein members of OmpR family. qRT-PCR analysis showed that the expression of OmpR503 gene was significantly enhanced by high salinity (90, 120). The expression of OmpR503 gene was also significantly increased at low temperature (0, 10 °C), whereas depressed at high temperature (30 °C). When the strain was subjected to combined stress (0 °C with a salinity of 90), the expression of OmpR503 gene was increased significantly, which was up to 3.0-fold. In Antarctica, freezing tolerance of psychrotrophic bacteria is often accompanied by tolerance to osmotic stress caused by a lack of free water, thus the cold inducibility of OmpR503 gene might help the strain adapt to the harsh environment more efficiently.

Low temperature is one of the major environmental challenges that Antarctic bacteria must face. Detailed studies of cold shock responses of cold-adapted microorganisms are still insufficient. Here, we cloned three cold shock protein (CSP) genes (Csp1137, Csp2039, and Csp2531) in the Antarctic bacterium Psychrobacter sp. G and their regulatory sequences were identified. The three CSPs were highly conserved with other known CspAs. qRT-PCR was performed to evaluate their expression characteristics under stress conditions, and the potential influence of regulatory sequences also was analyzed. The expression of Csp1137 was enhanced both by low (0, 10 °C) and high temperature (30 °C). The expression of Csp2039 was enhanced by low temperature (0 °C), but was lower than that of Csp1137. This can be explained by the absence in Csp2039 of the AT-rich UP element. Different from Csp1137, the expression of Csp2531 was inhibited by low temperature (0 °C), even with the presence of AT-rich UP element, and it was not sensitive to high temperature (30 °C). The expression of Csp1137 was enhanced by high salinity (90, 120), whereas that of Csp2531was enhanced by low salinity (0, 15). At 0 °C and a salinity of 15, the expression of Csp1137 was repressed initially, but then it increased greatly during the next 10 h. The expressions of Csp2039 and Csp2531 were repressed significantly under four different combinations of stress conditions. Our results showed that the role of the upstream regulation sequences were much more complex than previously thought. Also, gene expressions were also affected by the environmental salinity. These are helpful in further clarification of the adaptation mechanism of Psychrobacter sp. G.

A total of 69 strains of thermophilic bacteria were isolated from water, soil and sediment samples from three Indonesia’s hot spring areas (Pantai cermin, Kalianda and Banyu wedang) by using Minimal Synthetic Medium (MSM). The extreme thermophile Brevibacillus sp. PLI-1 was found to produce extracellular thermophilic alkaline protease with optimal activity at 70° and pH 8.0–9.0. The molecular weight of the protease was estimated to be around 56 kD by SDS-PAGE. The maximum activity of the protease was 26.54 U mL−1. The protease activity did not decrease after 30 min and still retained more than 70% of relative activity after 60 min at 70°C and pH 8.0. The ion Mg2+ was found to promote protease activity at both low and high concentrations, whereas Cu2+ and Zn2+ could almost completely inhibit the activity. Divalent cation chelator EDTA inhibited the enzyme activity by 55.06% ± 0.27%, while the inhibition caused by PMSF, Leupeptin, Pepstain A and Benzamidine were 66.78% ± 3.25%, 52.37% ± 0.25%, 62.47% ± 2.96% and 50.99% ± 0.24%, respectively. Based on these observations, the enzyme activity was conspicuously sensitive to the serine and cysteine protease inhibitors. All these results indicated that the protease isolated from the strain PLI-1 was a thermophilic protease and had a high-temperature stability and a pH stability.

Phytoplankton size structure plays a significant role in controlling the carbon flux of marine pelagic ecosystems. The mesoscale distribution and seasonal variation of total and size-fractionated phytoplankton biomass in surface waters. as measured by chlorophyll a (Chl a), was studied in the Southern Yellow Sea using data from four cruises during 2006-2007. The distribution of Chl a showed a high degree of spatial and temporal variation in the study area. Chl a concentrations were relatively high in the summer and autumn, with a mean of 142 and 1.27 mg m(-3), respectively. Conversely, in the winter and spring. the average Chl a levels were only 098 and 0.99 mg m(-3) Total Chl a showed a clear decreasing gradient from coastal areas to the open sea in the summer, autumn and winter cruises. Patches of high Chl a were observed in the central part of the Southern Yellow Sea in the spring due to the onset of the phytoplankton bloom. The eutrophic coastal waters contributed at least 68% of the total phytoplankton biomass in the surface layer. Picophytoplankton showed a consistent and absolute dominance in the central region of the Southern Yellow Sea (>40%) in all of the cruises, while the proportion of microphytoplankton was the highest in coastal waters The relative proportions of pico- and nanophytoplankton decreased with total biomass, whereas the proportion of the micro-fraction increased with total biomass. Relationships between phytoplankton biomass and environmental factors were also analysed. The results showed that the onset of the spring bloom was highly dependent on water column stability. Phytoplankton growth was limited by nutrient availability in the summer due to the strong thermocline. The combined effects of P-limitation and vertical mixing in the autumn restrained the further increase of phytoplankton biomass in the Surface layer. The low phytoplankton biomass in winter was caused by vertical dispersion due to intense mixing. Compared with the availability of nutrients. temperature did not seem to cause direct effects on phytoplankton biomass and its size structure. Although interactions of many different environmental factors affected phytoplankton distributions. hydrodynamic conditions seemed to be the dominant factor. Phytoplankton size structure was determined mainly by the size-differential capacity in acquiring resource. Short time scale events, such as the spring bloom and the extension of Yangtze River plume, can have substantial influences, both on the total Chl a concentration and on the size structure of the phytoplankton. (C) 2009 Elsevier Ltd. All rights reserved.

We compared four esterifiable enzymes. The lipase Novozym 435 possessed the highest activity for the conjugated linoleic acid esterification during the synthesis of triglycerides. The triglycerides were synthesized by esterification of glycerol and conjugated linoleic acid (CLA) in a no-solvent system using lipase catalysis. We investigated the effects of temperature, enzyme concentration, water content, and time on esterification. Enzyme and water concentrations of up to 1% of the total reaction volume and a system temperature of 60°C proved optimal for esterification. Similarly, when the esterification was carried out for 24 h, the reaction ratio improved to 94.11%. The esterification rate of the rotating screen basket remained high (87.28%) when the enzyme was re-used for the 5th time. We evaluated the substrate selectivity of lipase (NOVO 435) and determined that this lipase prefers the 10,12-octadacadienoic acid to the 9,11-octadecadienoic acid.

Here we report the draft genome sequence of Pseudoalteromonas strain A2, isolated from Arctic seawater in the pack-ice zone, which has high antioxidative activity against H 2 O 2 . The genomics information of this strain will facilitate the study of antioxidative mechanisms, cold adaptation properties, and evolution of this genus.

Preoperative acute hypervolemic hemodilution (AHHD) is a technique used in anesthesia to reduce the number of blood cells lost during intraoperative bleeding. The aim of the present study was to evaluate the effect of the hypervolemic hemodilution of 6% hydroxyethyl starch 130/0.4 on the EC50 of propofol at two clinical endpoints. A total of 20 patients undergoing AHHD following epidural anesthesia were studied, and 20 patients who did not receive hemodilution were used as a control group. All patients were American Society of Anesthesiologists grade I, aged 20–40 years and undergoing hip arthroplasty surgery. In the AHHD group, 10 ml/kg lactated Ringer's solution was infused over 20 min at the same time as the epidural test dose. The infusion was followed by the infusion of 6% hydroxyethyl starch 130/0.4 over 30 min. Patients in the control group received 10 ml/kg Ringer's solution over 50 min. Propofol was then delivered by a Diprifusor target-controlled infusion. The predicted blood and effect-site propofol concentrations were recorded at loss of consciousness (LOC) and return of consciousness (ROC). Probit analysis was used to estimate the values for predicted blood and effect-site concentrations at the two clinical endpoints. The results showed that the potency of propofol was decreased during AHHD. Compared with the controls, the predicted blood and effect-site concentrations of propofol at LOC were higher in patients of the hemodilution group, resulting in higher EC50 values (P=0.001 and 0.025, respectively). At ROC, the effect-site EC50 was 2.9 µg/ml [95% confidence interval (CI), 2.8–3.0] in hemodilution patients and 2.5 µg/ml (95% CI, 2.2–2.6) in control patients (P=0.001). With AHHD, the LOC time was significantly longer and the propofol dose was higher, while ROC times were comparable. In conclusion, AHHD increases the requirement for propofol at LOC and prolongs LOC time. Patients with AHHD recovered consciousness at higher effect-site concentrations of propofol. Thus, the induction dose of propofol should be increased during AHHD.

海面養殖の拡大により，多くの海産外来種が，社会経済あるいは生態系への影響を検討されることなく中国国内に持ち込まれている。そのうち，養殖用として 14 種を超える生物が日本から導入され，それらの大部分は現在では中国国内で定着していると考えられている。しかし，それらが中国の海洋生態系の多様性や在来種の生存に及ぼす影響についての知見は少ない。そのため，様々な外来種，とりわけ，今後導入される可能性があるかあるいは分布域を拡大する恐れのある外来種に関してのリスク評価並びに管理体制が求められている。（浜口昌巳 訳）

Here, we report the complete genome sequence of Psychrobacter sp. strain G, isolated from King George Island, Antarctica, which can produce lipolytic enzymes at low temperatures. The genomics information of this strain will facilitate the study of the physiology, cold adaptation properties, and evolution of this genus.

Objective: The purpose of the present study was to investigate the pharmacokinetics and efficacy of ropivacaine in Chinese patients by intra-articular administration after arthroscopic knee surgery, in order to assess the safety and efficacy. Patients and methods: 21 ASA I-II patients received a single-dose of ropivacaine 150 mg in a 20 ml intra-articular injection at the end of surgery. Plasma samples were collected prior to and after ropivacaine administration. Plasma concentrations of ropivacaine were measured by HPLC. Pharmacokinetic parameters were calculated using noncompartmental analysis. Population pharmacokinetic modeling was performed to yield estimates of clearance, volume of distribution, and absorption rate constant. An analysis of covariates on the pharmacokinetic parameters was also carried out. Pain assessments were made using a verbal rating scale at intervals of 2, 4, 8, 12, 24, 36, 48 and 72 hours after surgery. Results: The results show that the peak plasma concentrations occurred at an average of 0.93 ± 0.56 h (0.25 - 2 h), with a mean of 0.91 ± 0.4 mg/l (range 0.35 - 1.54 mg/l). The peak plasma concentrations and the times to reach the peak plasma concentration exhibited a marked variability among the subjects. All concentrations were well below the estimated toxic threshold (2.2 mg/l). No patient experienced adverse events that may have been related to ropivacaine administration. The intra-articular use of ropivacaine provided excellent control of pain after knee arthroscopy. Conclusion: Ropivacaine 150 mg provided satisfactory postoperative pain relief and can be safely administered by intraarticular injection in Chinese patients after arthroscopic knee surgery and the pharmacokinetic profiles of ropivacaine exhibited marked variability among the subjects. The high variability of pharmacokinetic profiles in this study may be caused by gender and body weight.

Statistical experimental designs were applied to optimize cold-active lipase production by the psychrophilic bacterium Moritella sp. 2-5-10-1. First, a Plackett–Burmen design (PBD) was used to evaluate the significant effects of various fermentation parameters. The results indicated that soybean meal, temperature, and Tween-80 had significant influences on lipase production. The levels of these variables were optimized subsequently using central composite design (CCD). A quadratic regression model of cold-active lipase production was built, and verification experiments confirmed its validity. On subsequent scale-up in a 10-L bioreactor using optimized conditions, cold-active lipase production (30.56 U/mL) was obtained. The results clearly indicated that the model was adequate even on a large scale. To our knowledge, this is the first report of statistical optimization of cold-active lipase production by a psychrophilic bacterium.

Temperature and salinity fluctuations are two of the most important factors affecting the growth of polar bacteria. In an attempt to better understand the function of heat-shock proteins (HSPs) in the adaptive mechanisms of the Antarctic psychrotrophic bacterium Psychrobacter sp. G to such conditions, genes Hsp845, Hsp2538, Hsp2666, and Hsp2667 were cloned on the basis of the draft genome. The expression characteristics of these HSP genes under different stress conditions were analyzed by the qRT-PCR method. Expression of Hsp845 and Hsp2667 was inhibited significantly by low temperature (0 and 10 °C, respectively). There was no difference of expression when Hsp2538 and Hsp2666 were exposed to 0 °C but the expression of Hsp2666 was inhibited when exposed to 10 °C. Expression of Hsp2538 and Hsp2667 was not sensitive but expression of Hsp845 and Hsp2666 was increased at low salinity (0 and 15, respectively). Expression of the four HSP genes was enhanced at high salinity (90 and 120) and at high temperature independent of salinity. By contrast, low temperature had no significant effect independent of salinity.

The ability of Antarctic ice algae, Chlamydomonas sp. ICE-L, to survive and proliferate at low temperature and high salinity implies that they have overcome key barriers inherent in Antarctic environments. A full-length complementary DNA (cDNA) sequence of omega-3 fatty acid desaturase, designated CiFAD3, was isolated via reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends methods. The full-length of CiFAD3 cDNA contained an open reading frame of 1,302 bp with 5′-terminal untranslated region (UTR) of 36 bp and 3′-terminal UTR of 507 bp encoding a fatty acid desaturase protein of 434 amino acids. Sequence alignment and phylogenetic analysis showed that the gene was homologous to known chloroplastic omega-3 fatty acid desaturase. Meanwhile, CiFAD3 sequence showed typical features of membrane-bound desaturase such as three conserved histidine boxes along with four membrane spanning regions that were universally present among plant desaturases. Under different stress conditions, messenger RNA (mRNA) expression levels of CiFAD3 were measured by quantitative RT-PCR. The results showed that both temperature and salinity could motivate the upregulation of CiFAD3 expression. The mRNA accumulation of CiFAD3 increased 2.6-fold at 0°C and 1.8-fold at 12°C compared to the algae at 6°C. Similarly, mRNA expression levels of CiFAD3 increased 3.8-fold after 62‰ NaCl treatment for 2 h. However, CiFAD3 mRNA expression levels were partially decreased after UV radiation. These data suggest that CiFAD3 is the enzyme responsible for the omega-3 fatty acid desaturation involved in ice algae Chlamydomonas sp. ICE-L acclimatizing to cold temperature and high salinity in Antarctic environment.

A cDNA encoding heat shock protein 70 of Antarctic ice algae Chlamydomonas sp. ICE-L (designated as CiHsp70) was identified by RT-PCR and rapid amplification of cDNA ends approaches. The full-length cDNA of CiHsp70 was 2,232 bp, consisting of a 5'-terminal untranslated region (UTR) of 76 bp, a 3'-terminal UTR of 203 bp with a poly (A) tail, and an open reading frame of 1,953 bp. The CiHsp70 cDNA encoded a polypeptide of 651 amino acids with an ATPase domain of 388 amino acids, the substrate peptide binding domain of 246 amino acids and a C-terminus domain of 17 amino acids. The inducible CiHsp70 cDNA was highly homologous to other plant cytosolic Hsp70 genes and clustered together with green algae and higher plant rather than brown algae, diatom and Cryptophyta. Antarctic ice algae were treated with different stress conditions and messenger RNA (mRNA) expression levels of CiHsp70 were quantified by quantitative RT-PCR. The results showed that both cold and heat shock treatments could stimulate CiHsp70 mRNA expression. Meanwhile, CiHsp70 mRNA expression level increased 2.9-fold in response to UV-B radiation for 6 h, while the expression levels of CiHsp70 were remarkably increased after removing the UV-B radiation and immediately providing additional 6 h visible light. Furthermore, treating with 62 or 93 per thousand NaCl for 2 h, CiHsp70 mRNA expression level increased 3.0- and 2.1-fold, respectively. Together, our observations revealed that CiHsp70 as a molecular chaperone might play an important role in Antarctic ice algae Chlamydomonas sp. ICE-L acclimatizing to polar environment.

A psychrotrophic petroleum-degrading bacterium Pseudoalteromonas sp. P29 was isolated from marine sediment, which was collected during 2nd Chinese Arctic Scientific Expedition. The phenotypic character and biodegradation efficiency on mixed oil or vacuum oil were tested at low temperature. The strain Pseudoalteromonas sp. P29 grew in a range of temperature from 5 to 35 degrees C and the optimum temperature was 25 degrees C. Gas chromatography analysis indicated that the strain might preferentially metabolize shorter-chain alkanes. The biodegradation efficiency were nearly 90 and 80%, respectively, after incubation at 5 degrees C for 28 days in the mineral medium supplement with mixed oil or vacuum oil as the sole carbon and energy source. The results showed a possible exploitation of the strain in future biotechnological processes especially in cold contaminated environments.

Calcium-activated potassium channels on plasma membrane enable potassium influx into the cell with ensuing changes in plasma membrane potential and consequent effects on cellular metabolic functions. Recently, this potassium channel was reported to regulate the cellular responses of mammalian immune cells. We have postulated the presence of such a channel in fish immune cells and its potential role in immunoregulation in fish. Employing specific primers and RNA template, we cloned a segment of a novel gene from turbot blood sample and subsequently obtained a full cDNA sequence using RACE approaches. Bioinformatic analysis revealed structural and phylogenetic characteristics of a novel small conductance calcium-activated potassium channel gene, we called TSKCa, which exhibits homologous domains to other species particularly in the transmembrane regions. Full-length TSKCa cDNA is 1698 bp with a 1632 bp open reading frame encoding a protein of 544 amino acids. TSKCa gene is expressed in majority of the tested organs and tissues of turbot. To assess the postulated immune function of TSKCa, we infected turbot with the pathogen Vibrio anguillarum. Here, semi-quantitative RT-PCR analysis demonstrated increased mRNA expression of TSKCa in head kidney, spleen and blood, indicating an important role of TSKCa in these organ tissues that mediate the immune defense response of turbot. In contrast, there was less change in expression in the turbot intestines and liver which were less implicated in the immune response in present study.

A gene (lipP, 837 bp in length) coding for a cold-adapted lipase of psychrophilic bacterium Moritella sp. 2-5-10-1 isolated from Antarctic region was cloned and sequenced in this study. The deduced amino acid sequence revealed a protein of 278 amino acid residues with a molecular mass of 30,521. The primary structure of the lipase deduced from the nucleotide sequence showed consensus pentapeptide containing the active serine [Gly-Trp-Ser-Leu-Gly] and a conserved His-Gly dipeptide in the N-terminal part of the enzyme. These sequences were involved in the lipase active site conformation. Structure factors that would allow proper enzyme flexibility at low temperatures were discussed. It was suggested that the changes in the primary structure of the psychrophilic lipases compared to the thermophilic ones could account for their ability to catalyze lipolysis at temperatures close to 0 degrees C. For expression, the sequence corresponding to the cold-adapted lipase of strain 2-5-10-1 was subcloned into the pET-28a expression vector to construct a recombinant lipase protein. Expression of the lipase by Escherichia coli BL21 (DE3) cells was observed as clear halos on 1% (vol/vol) tributyrin upon induction with IPTG at 25 degrees C.