The relationship between soft-bottom macrobenthic communities and environmental variables off Ningjin,eastern Shandong Peninsula

Macrobenthic infauna and associated environmental factors influencing the benthic community in the eastern coastal region of Shandong Peninsula were analyzed in four seasonal surveys from January 2007 to October 2007 （30 stations in winter,20 stations in other three seasons）,in order to understand the community structure and the factors unfluencing the benthic distribution.PRIMER 6.0 and SPSS 15.0 software packages were adopted to analyze the environmental and macrobenthic data.The results show that there were 260 macrobenthic species in total collected from the research region.The composition of species is： Polychaeta （94 species）,Crustacea （75）,Mollusca （56） and Echinoderm （12）,among which,only 23 species were common species in the cruises of every season.The dominant species varied from season to season;however,the polychaete species Paralacydonia paradoxa Fauvel and Echinoderm species Amphioplus japonicus （Matsumoto） were always present year-round.The abundance and biomass of the macrobenthos in the research region were variable from season to season.The results of CLUSTER and MDS analysis show that the similarities of macrobenthic structures between the stations were low;most of the similarities were at about 30% of similarity value,only two stations were up to 70%.In accordance with the similarity values of the macrobenthic structures,we divided the 20 stations into five groups by arbitrary similarity level of 30%.The ABC curve indicates that the marcofauna communities in the research region had not been disturbed massively,except two stations,SB1 and SB3.Ecologically,benthos were controlled by a combination of factors such as salinity,phytoplankton,zooplankton,SiO 3Si and temperature,and no single factor could be considered as an ecological master factor.

Nitrogen Biological Cycle Characteristics of Seepweed(Suaeda salsa) Wetland in Intertidal Zone of Huanghe(Yellow) River Estuary

From April 2008 to November 2009, the nitrogen （N） cycle of plant-soil system in seepweed （Suaeda salsa） wetland in the intertidal zone of the Huanghe （Yellow） River estuary was studied. Results showed that soil N had sig- nificant seasonal fluctuations and vertical distribution, and the net N mineralization rates in topsoil were significantly different in growing season （p 〈 0.01）. The N/P ratio （9.87 ±1.23） of S. salsa was less than 14, indicating that plant growth was limited by N. The N accumulated in S. salsa litter at all times during decomposition, which was ascribed to the N immobilization by microbes from the environment. Soil organic N was the main N stock of plant-soil system, accounting for 97.35% of the total N stock. The N absorption and utilization coefficients of S. salsa were very low （0.0145 and 0.3844, respectively）, while the N cycle coefficient was high （0.7108）. The results of the N turnovers among compartments of S. salsa wetland showed that the N uptake amount of aboveground part and root were 7.764 g/m2and 4.332 g/m2, respectively. The N translocation amounts from aboveground part to root and from root to soil were 3.881 g/m2 and 0.626 g/m2, respectively. The N translocation amount from aboveground living body to litter was 3.883 g/m2, the annual N return amount from litter to soil was more than 0.125（-） g/m2 （minus represented immobili- zation）, and the net N mineralization amount in topsoil （0-15 cm） in growing season was 1.190 g/m2. The assessment of N biological cycle status orS. salsa wetland indicated that N was a very important limiting factor and the ecosystem was situated in unstable and vulnerable status. The S. salsa was seemingly well adapted to the low-nutrient status and vulnerable habitat, and the N quantitative relationships determined in the compartment model might provide scientific base for us to reveal the special adaptive strategy orS. salsa to the vulnerable habitat in the following studies.

Anthropogenic impacts on hyperbenthos in the coastal waters of Sishili Bay,Yellow Sea

A survey on the hyperbenthic communities was carried out in the coastal waters of Sishili Bay in the Yellow Sea in July 2009,to investigate the impact of putative anthropogenic activities related to the presence of a sewage outfall,a harbor and an aquaculture site on the benthic ecosystem.An Agassiz net trawl was used to collect hyperbenthos at 10 sampling stations.Species,diversity,abundance,and biomass were analyzed against water sample data and historical data obtained from records from nearby Yantai City.Fifty-two species were identified in the region,of which Crustacea were the most abundant taxon,followed by Echinodermata.Dominant species included five crustaceans,two echinoderms,one mollusk and one fish species.The results of a BIOENV analysis show that the concentrations of NO3-N and Chl-a were slightly positively correlated with hyperbenthic community structure,while other factors were negatively correlated,including sediment grain size and the percentage of TN and TOC in sediment.Abundance- Biomass Comparison(ABC method) curves indicate that the hyperbenthos in Sishili Bay had been disturbed by putative sources of human activities.Eight out of 10 stations were classified as "moderately perturbed" to "perturbed".

Molecular cloning, characterization and expression profiles of thioredoxin 1 and thioredoxin 2 genes in Mytilus galloprovincialis

Thioredoxin （Trx） proteins are involved in many biological processes especially the regulation of cellular redox homeostasis. In this study, two Trx cDNAs were cloned from the mussel Mytilus galloprovincialis using rapid amplification of cDNA ends-polymerase chain reaction （RACE-PCR）. The two cDNAs were named MgTrxl and MgTrx2, respectively. The open reading frames of MgTrxl and MgTrx2 were 318 and 507 base pairs （bp） and they encoded proteins of 105 and 168 amino acids with estimated molecular masses of 11.45 and 18.93 kDa, respectively. Sequence analysis revealed that both proteins possessed the conserved active site dithiol motif Cys-Gly-Pro-Cys. In addition, MgTrx2 also possessed a putative mitochondrial targeting signal suggesting that it is located in the mitochondria. Quantitative real-time polymerase chain reaction （qPCR） revealed that both MgTrxl and MgTrx2 were constitutively expressed in all tissues examined. The MgTrxl transcript was most abundant in hemocytes and gills, whereas the MgTrx2 transcript was most abundant in gonad, hepatopancreas, gill and hemocytes. Following Vibrio anguillarum challenge, the expression of MgTrxl was up-regulated and reached its peak, at a value 10-fold the initial value, at 24 h. Subsequently, expression returned back to the original level. In contrast, the expression level of MgTrx2 was down-regulated following bacterial stimulation, with one fifth of the control level evident at 12 h post challenge. These results suggest that MgTrxl and MgTrx2 may play important roles in the response ofM. galloprovincialis to bacterial challenge.

Application of Stable Isotope Techniques in Studies of Carbon and Nitrogen Biogeochemical Cycles of Ecosystem

Stable isotope techniques have been proved useful as tools for studying the carbon (C) and nitrogen (N) biogeochemical cycles of ecosystem. This paper firstly introduced the basic principles and the distribution characteristics of stable isotope, then reviewed the recent advances and applications of stable isotope in the C and N biogeochemical cycles of ecosystem. By applying the 13 C natural abundance technique, ecologists are able to understand the photosynthetic path and CO 2 fixation of plants, the CO 2 exchange and C balance status of ecosystem, the composition, distribution and turnover of soil organic C and the sources of organic matter in food webs, while by using the 13 C labeled technique, the effects of elevated CO 2 on the C processes of ecosystem and the sources and fate of organic matter in ecosystem can be revealed in detail. Differently, by applying the 15 N natural abundance technique, ecologists are able to analyze the biological N 2 -fixation, the N sources of ecosystem, the N transformation processes of ecosystem and the N trophic status in food webs, while by using the 15 N labeled technique, the sources, transformation and fate of N in ecosystem and the effects of N input on the ecosystem can be investigated in depth. The applications of both C and N isotope natural abundance and labeled techniques, combined with the elemental, other isotope ( 34 S) and molecular biomarker information, will be more propitious to the investigation of C and N cycle mechanisms. Finally, this paper concluded the problems existed in current researches, and put forward the perspective of stable isotope techniques in the studies on C and N biogeochemical cycles of ecosystem in the future.

The influence of salinity on toxicological effects of arsenic in digestive gland of clam Ruditapes philippinarum using metabolomics

Ruditapes philippinarum,a clam that thrives in intertidal zones of various salinities,is a useful biomonitor to marine contaminants.We investigated the influence of dilution to 75% and 50% of normal seawater salinity(31.1) on the responses of the digestive gland of R.philippinarum to arsenic exposure(20 μg/L),using nuclear magnetic resonance(NMR)-based metabolomics.After acute arsenic exposure for 48 h,salinity-dependent differential metabolic responses were detected.In normal seawater,arsenic exposure increased the concentrations of branched-chain amino acids,and of threonine,proline,phosphocholine and adenosine,and it decreased the levels of alanine,hypotaurine,glucose,glycogen and ATP in the digestive glands.Differential changes in metabolic biomarkers observed at lower salinity(～23.3) included elevation of succinate,taurine and ATP,and depletion of branched-chain amino acids,threonine and glutamine.Unique effects of arsenic at the lowest salinity(～15.6) included down-regulation of glutamate,succinate and ADP,and up-regulation of phosphocholine.We conclude that salinity influences the metabolic responses of this clam to arsenic.

Temporal and spatial variations in the distribution of macroalgal communities along the Yantai coast, China

To explore the impact of environmental variables on macroalgal communities, their temporal and spatial distributions were examined along the Yantai coast, China between April 2010 and March 2011. Macroalgae sampling was conducted monthly at four sites along the coast: Jiahe River estuary, Zhifu Island, Fisherman's Wharf, and Yangma Island. The species composition and abundance, and their relationship with environmental variables were assessed. Along the Yantai coast, 35 macroalgae species were identifi ed, including 24 Rhodophyta spp., 7 Chlorophyta, and 4 Phaeophyta spp. Highest species numbers were recorded in the summer at all sampling sites, except in the Jiahe River estuary. Macroalgae biomass was the greatest in the summer. Year-round, the highest species number and dry biomass recorded at Fisherman's Wharf and Yangma Island was attributed to the substrate type. In summer, Ulva pertusa Kjellman was the dominant species identifi ed along the Yantai coast, which indicates a risk of macroalgae blooms. Our results show that seawater temperature and nutrients appear to signifi cantly affect the temporal and spatial patterns of macroalgal abundance along the Yantai coast. The effects of environmental variables on the macroalgae on the Yantai coast need further study.

Glutathione S-transferase(GST) gene expression profiles in two marine bivalves exposed to BDE-47 and their potential molecular mechanisms

Glutathione S-transferases （GSTs） are phase II enzymes that facilitate the detoxification of xenobioties and play important roles in antioxidant defense. We investigated the expression patterns of seven Venerupis philippinarum GSTs （VpGSTs） and four Mytilus galloprovincialis GSTs （MgGSTs） following exposure to BDE-47. Differential expressions of the seven VpGSTs and four MgGSTs transcripts were observed, with differences between the hepatopancreas and gills. Among these GSTs, the sigma classes （VpGSTS1, VpGSTS2, VpGSTS3, MgGST1, and MgGST3） were highly expressed in response to BDE-47 exposure, demonstrating their potential as molecular biomarkers for environmental biomonitoring studies. We obtained the three-dimensional crystal structures of VpGSTs and MgGSTs by homologous modeling. A model to elucidate the binding interactions between the ligands and receptors was defined by molecular docking, Hydrophobic and n were the most often observed interactions between BDE-47 and the GSTs.

Effect of cadmium on the defense response of Pacific oyster Crassostrea gigas to Listonella anguillarum challenge

Heavy metal pollution can affect the immune capability of organisms. We evaluated the effect of cadmium （Cd） on the defense responses of the Pacific oyster Crassostrea gigas to Listonella anguillarum challenge. The activities of several important defensive enzymes, including superoxide dismutase （SOD）, glutathione peroxidase （GPx）, acid phosphatase （ACP）, Na＋, K＋-ATPase in gills and hepatopancreas, and phenoloxidase-like （POL） enzyme in hemolymph were assayed. In addition, the expression levels of several genes, including heat shock protein 90 （IrtSP9~））, metallothionein （MT）, and bactericidal/permeability increasing （BPI） protein were quantified by fluorescent quantitative PCR. The enzyme activities of SOD, ACP, POL, and GPx in hepatopancreas, and the expression of HSP90 were down-regulated, whereas GPx activity in the gill, Na＋, K＋-ATPase activities in both tissues, and MT expression was increased in Cd- exposed oysters post L. anguillarum challenge. However, BPI expression was not significantly altered by co-stress of L. anguillarum infection and cadmium exposure. Our results suggest that cadmium exposure alters the oysters＇ immune responses and energy metabolism following vibrio infection.

Runoff variation affected by precipitation change and human activity in Hailiutu River basin,China

For recent years,runoff generation and hydrological processes in Hailiutu River basin have been greatly changed by climate change and human activity,especially water and soil conservation construction.In this study,the trends in precipitation,evapotranspiration(ET)and river runoff as well as the effects of precipitation change and human activity on runoff variation have been studied.The results showed that during 1960-2000,annual precipitation and river runoff,monthly precipitation and ET in September and October as well as monthly runoff in all months showed a significant decrease.In addition,peak flow and base flow had a large decrease.Under the joint influence of precipitation change and human activity,the mean annual runoff decreased by 35 million m3 from the baseline period(1960-1985)to the change period(1986-2000),which accounted for 60.9%and 39.1%of the total runoff decrease,respectively.Precipitation change played a primary role in the decrease of annual runoff whereas human activity,particularly water and soil conservation construction,also had remarkable impacts on runoff variation.

Homologous cloning, characterization and expression of a new halophyte phytochelatin synthase gene in Suaeda salsa

The halophyte Suaeda salsa can grow in heavy metal-polluted areas along intertidal zones having high salinity.Since phytochelatins can effectively chelate heavy metals,it was hypothesized that S.salsa possessed a phytochelatin synthase(PCS) gene.In the present study,the cDNA of PCS was obtained from S.salsa(designated as SsPCS) using homologous cloning and the rapid amplification of cDNA ends(RACE).A sequence analysis revealed that SsPCS consisted of 1 916 bp nucleotides,encoding a polypeptide of 492 amino acids with one phytochelatin domain and one phytochelatin C domain.A similarity analysis suggested that SsPCS shared up to a 58.6%identity with other PCS proteins and clustered with PCS proteins from eudicots.There was a new kind of metal ion sensor motif in its C-terminal domain.The SsPCS transcript was more highly expressed in elongated and fibered roots and stems(P<0.05) than in leaves.Lead and mercury exposure significantly enhanced the mRNA expression of SsPCS(P<0.05).To the best of our knowledge,SsPCS is the second PCS gene cloned from a halophyte,and it might contain a different metal sensing capability than the first PCS from Thellungiella halophila.This study provided a new view of halophyte PCS genes in heavy metal tolerance.

Molecular cloning and differential expression patterns of sigma and omega glutathione S-transferases from Venerupis philippinarum to heavy metals and benzo[a]pyrene exposure

Glutathione S-transferases (GSTs) are a class of enzymes that facilitate the detoxification of xenobiotics, and also play important roles in antioxidant defense. We identified two glutathione S-transferase isoforms (VpGSTS, sigma GST; VpGSTO, omega GST) from Venerupis philippinarum by RACE approaches. The open reading frames of VpGSTS and VpGSTO were of 612 bp and 729 bp, encoding 203 and 242 amino acids with an estimated molecular mass of 22.88 and 27.94 kDa, respectively. The expression profiles of VpGSTS and VpGSTO responded to heavy metals and benzo[a]pyrene (B[a]P) exposure were investigated by quantitative real-time RT-PCR. The expression of VpGSTS and VpGSTO were both rapidly up-regulated, however, they showed differential expression patterns to different toxicants. Cd displayed stronger induction of VpGSTS expression with an approximately 12-fold increase than that of VpGSTO with a maximum 6.4-fold rise. Cu exposure resulted in similar expression patterns for both VpGSTS and VpGSTO. For B[a]P exposure, the maximum induction of VpGSTO was approximately two times higher than that of VpGSTS. Altogether, these findings implied the involvement of VpGSTS and VpGSTO in host antioxidant responses, and highlighted their potential as a biomarker to Cd and B[a]P exposure.

Macroalgae Blooms and their Effects on Seagrass Ecosystems

Seagrass decline caused by the macroalgae blooms is becoming a common phenomenon throughout temperate and tropical regions. We summarized the incidence of macroalgae blooms throughout the world and their impact on seagrass beds by direct and indirect ways. The competition for living space and using resources is the most direct effect on seagrass beds when macroalgae are blooming in an aquatic ecosystem. The consequence of macroalgae blooms(e.g., light reduction, hypoxia, and decomposition) can produce significant indirect effects on seagrass beds. Light reduction by the macroalgae can decrease the growth and recruitment of seagrasses, and decomposition of macroalgae mats can increase the anoxic and eutrophic conditions, which can further constrict the seagrass growth. Meanwhile, the presence of seagrass shoots can provide substrate for the macroalgae blooms. Controlling nutrient sources from the land to coastal waters is a general efficient way for coastal management. Researching into the synergistical effect of climate change and anthropognic nutrient loads on the interaction between searsasses and macroalgae can provide valuable information to decrease the negative effects of macroalgae blooms on seagrasses in eutrophic areas.

Manila clam Venerupis philippinarum as a biomonitor to metal pollution

The Manila clam Venerupis philippinarum is a good biomonitor/bioindicator to marine metal pollution and is frequently used in aquatic toxicology. Two dominant pedigrees (white and zebra) of clam are distributed in the Bohai Sea; however, little attention has been paid to potential biological differences between these two pedigrees. In this study, we tested the sensitivity of both pedigrees to marine metal (cadmium and zinc) pollution biomonitoring and marine environmental toxicology. Results demonstrate significant biological differences in gills of white and zebra clams based on metabolic profiles and antioxidant enzyme activities. In addition, we found that hypotaurine, malonate and homarine were relatively high in white clam gills, while alanine, arginine, glutamate, succinate, 4-aminobutyrate, taurine and betaine were high in zebra clam gills. Zebra clam gills were also more sensitive to a mixture of Cd and Zn, as shown by antioxidant enzyme activities and metabolic profiles, but white clam gills could accumulate more Zn. Therefore, we suggest that the white pedigree can be used as a biomonitor to marine Zn pollution, whereas the zebra pedigree can be used for toxicology studies on Cd and Zn mixed pollution.

Proteomic responses induced by metal pollutions in oysters Crassostrea sikamea

There exist severe metal pollutions along the Jiulongjiang estuary in South China. In order to unravel the biological eff ects caused by metal pollutions, proteomic responses were investigated by two-dimensional electrophoresis-based proteomics in oysters Crassostrea sikamea from metal pollution sites, Jinshan (JS) and Baijiao (BJ), and a relatively clean site, Jiuzhen (JZ), along the Jiulongjiang estuary. Results indicated that metal pollutions mainly induced cellular injuries, oxidative and immune stresses, and disturbed ion homeostasis in oysters C . sikamea from both JS and BJ sites via diff erential pathways. Furthermore, metal pollution enhanced transcriptional initiation in oysters from JS site. In addition, the Cu and Fe pollution might be indicated by the 78 kDa glucose regulated protein and ferritin GF1 in oysters C . sikamea , respectively. The study confirms that proteomics is a promising approach to characterize the underlying mechanisms of responses to metal pollution in oysters.

Wave steepness retrieved from scatterometer data in a genetic algorithm

Wave steepness is an important characteristic of a high sea state, and is widely applied on wave propagations at ports, ships, offshore platforms, and CO2 circulation in the ocean. Obtaining wave steepness is a difficult task that depends heavily on theoretical research on wavelength distribution and direct observations. Development of remote-sensing techniques provides new opportunities to study wave steepness. At present, two formulas are proposed to estimate wave steepness from QuikSCAT and ERS-1/2 scatterometer data. We found that wave steepness retrieving is not affected by radar band, and polarization method, and that relationship of wave steepness with radar backscattering cross section is similar to that with wind. Therefore, we adopted and modified a genetic algorithm for relating wave steepness with radar backscattering cross section. Results show that the root-mean-square error of the wave steepness retrieved is 0.005 in two cases from ERS-1/2 scatterometer data and from QuikSCAT scatterometer data.

Characteristics of the Bohai Sea oil spill and its impact on the Bohai Sea ecosystem

In this paper, ENVISAT ASAR data and the Estuary, Coastal and Ocean Model was used to analyze and compare characteristics of the Bohai Sea oil spill. The oil slicks have spread from the point of the oil spill to the east and north-western Bohai Sea. We make a comparison between the changes caused by the oil spill on the chlorophyll concentration and the sea surface temperature using MODIS data, which can be used to analyze the effect of the oil spill on the Bohai Sea ecosystem. We found that the Bohai Sea oil spill caused abnormal chlorophyll concentration distributions and red tide nearby area of oil spill.

Nitrogen cycle of a typical Suaeda salsa marsh ecosystem in the Yellow River estuary

The nitrogen（N） biological cycle of the Suaeda salsa marsh ecosystem in the Yellow River estuary was studied during 2008 to 2009.Results showed that soil N had significant seasonal fluctuations and vertical distribution.The N/P ratio（15.73±1.77） of S.salsa was less than 16,indicating that plant growth was limited by both N and P.The N absorption coefficient of S.salsa was very low（0.007）,while the N utilization and cycle coefficients were high（0.824 and 0.331,respectively）.The N turnover among compartments of S.salsa marsh showed that N uptake from aboveground parts and roots were 2.539 and 0.622 g/m2,respectively.The N translocation from aboveground parts to roots and from roots to soil were 2.042 and 0.076 g/m2,respectively.The N translocation from aboveground living bodies to litter was 0.497 g/m2,the annual N return from litter to soil was far less than 0.368 g/m2,and the net N mineralization in topsoil during the growing season was 0.033 g/m2.N was an important limiting factor in S.salsa marsh,and the ecosystem was classified as unstable and vulnerable.S.salsa was seemingly well adapted to the low-nutrient status and vulnerable habitat,and the nutrient enrichment due to N import from the Yellow River estuary would be a potential threat to the S.salsa marsh.Excessive nutrient loading might favor invasive species and induce severe long-term degradation of the ecosystem if human intervention measures were not taken.The N quantitative relationships determined in our study might provide a scientific basis for the establishment of effective measures.

Potentiometric sensor for sensitive and selective detection of heparin

A polymeric membrane ion-selective electrode for determination of heparin is described in this paper.Protamine is incorporated into the organic membrane phase and functions as sensing element for selective recognition of heparin.The proposed membrane electrode exhibits high selectivity for heparin over lipophilic anions such as thiocyanide and salicylate.The potentiometric response to the concentration of heparin is Unear in the range of 0.01-0.4 U/mL and a lower detection limit of 0.005 U/mL can be achieved.