A non-traditional fuzzy quantification method is presented in the modeling of an extreme significant wave height. First, a set of parametric models are selected to fit time series data for the significant wave height and the extrapolation for extremes are obtained based on high quantile estimations. The quality of these results is compared and discussed. Then, the proposed fuzzy model, which combines Poisson process and generalized Pareto distribution (GPD) model, is applied to characterizing the wave extremes in the time series data. The estimations for a long-term return value are considered as time-varying as a threshold is regarded as non-stationary. The estimated intervals coupled with the fuzzy theory are then introduced to construct the probability bounds for the return values. This nontraditional model is analyzed in comparison with the traditional model in the degree of conservatism for the long-term estimate. The impact on the fuzzy bounds of extreme estimations from the non stationary effect in the proposed model is also investigated.

The major and trace elements in 110 surface sediment samples collected from the middle of the Bay of Bengal (mid-Bay of Bengal) are analyzed to investigate provenance. Si levels are highest, followed by Al, and the distributions of these two elements are identical. The average CIA^* (chemical index of alteration) value is 72.07, indicating that the degree of weathering of the sediments in the study area is intermediate between those of sediments of the Himalayan and Indian rivers. Factor analyses and discrimination function analyses imply that the two main provenances are the Himalayan and the Indian continent. The inverse model calculation of the Ti-normalized element ratios of the Bay of Bengal sediments indicate an estimated average contribution of 83.5% and 16.5% from the Himalayan and peninsular Indian rivers to the study area, respectively. The Himalayan source contributes more sediment to the eastern part of the study area, whereas the western part receives more sediment from the Indian Peninsula than did the eastern part. The primary mechanisms for deposition of sediments in the study area are the transport of Himalayan matter by turbidity currents and river-diluted water and the transport of Indian matter to the study area by a surface circulation in the Bay of Bengal, particularly the East India Coastal Current.

Quantitative evaluation of the copepod feeding process is critical for understanding the functioning of marine food webs, as this provides a major link between primary producers and higher trophic levels. In this study, a molecular protocol based on quantitative polymerase chain reaction (qPCR) targeting 18S rDNA was developed and used to investigate the feeding and digestion rates of the copepod Acartia erythraea in a laboratory experiment using microalgae Thalassiosira weissflogii, Prorocentrum shikokuense, and Alexandrium catenella as prey. Although offered an equal encounter rate based on biovolume, prey uptake varied substantially among the three algal species, with the ingestion rate (IR) and digestion rate (DR) of A. erythraea differing significantly (P< 0.001) based on both cell counting and qPCR detection.Acartia erythraea showed the highest IR (2.79 × 10⁴ cells/(ind.·h)) and DR (2.43 × 10⁴ cells/(ind.·h)) on T. weissflogii, and the lowest amounts of ingested P. shikokuense were detected. The highest assimilation rate (~90.64%, IR/DR) was observed in copepods fed with P. shikokuense. The qPCR method used here can help determine the digestion rate and assimilation rate of copepods by detecting cells remaining in the gut hence providing the possibility to examine trophic links involving key species in the marine ecosystem. Our results indicate that A. erythraea has diet-specific feeding performance in different processes, and a quantitative assessment of copepod feeding is needed to accurately determine its functional role in the energy and matter uptake from marine food webs.

The distribution of nitrate,nitrite,ammonia,phosphate and silicate in pore-water and their exchange between sediments and overlying waters (benthic flux) were determined at nine locations on the shelve of Huanghai and East China Seas.On the basis of the redox potential of sediments and nutrients distributions in the pore-waters,it is found that the benthic sediments are being in a suboxic to anoxic environment in the Huanghai and East China Seas.The nutrients distribution in the pore-waters is mainly controlled by the sediment environment,and ammonia is the major inorganic nitrogen in the pore-waters.On the basis of benthic fluxes of untrients calculated using Fick's first law,there is remarkable efflux of ammonia,dissolved inorganic nitrogen(DIN),phosphate and silicate from the sediments to the overlying waters in the study area,and their benthic fluxes are 299.3~2 214.8,404.4~2 159.5,5.5~18.8 and 541.3~1 781.6 μmol/(m²·d) respectively,and perhaps they are the major source of dissolved inorganic nitrogen,phosphate and cate for the overlying water.At most stations,the nitrate flux was from the overlying waters to the sediments,which suggests that suboxic organic matter decomposition via denitrification is dominated in the most area of Huanghai and East China Seas.High benthic fluxes appearing in the coastal area and relatively low benthic fluxes occurring in the shelf area are found and are consistent with primary productivity zoning in the study area.On the other hand,the ammonia flux displays an exponential decrease with water depth increase and an exponential increase with the bottom water temperature.However,others do not display this trend.

The quantification of transparent exopolymer particles (TEP) by colorimetric method is of large error and low repeatability, one major reason of which is related to the absence of clear definition and evaluation for part steps of the original method. It is obscure that the 80% sulfuric acid solution, acted as the extraction solution in the determination of TEP, is prepared based on a volume ratio or mass ratio. Furthermore, the change of solubility of recently available Gum Xanthan (GX) from the market means that the original protocol is no longer applicable, and the grinding of GX stock solution with a tissue grinder is replaced by shaking with a rotating shaker in the study to prevent the excessive dissolution of GX. We found that different preparation techniques could result in the varied concentrations of 80% H₂SO₄. The duration of shaking during the preparation of standard solution significantly affected the slope of the calibration curve, which caused different correction results of TEP. The impacts of different extraction solution concentrations and shaking time of GX solution on the quantification of TEP were investigated based on the field sampling and laboratory analysis. The extraction capacities of H₂SO₄ with different concentrations for Alcian Blue were distinct, but had limited effect on the final measuring result of TEP. The change of the standard curve slope came along with the variation of shaking time, which markedly altered the detection limit and calibration result, and the extended shaking time was in favor of the determination of low-concentration TEP. It was suggested that the extraction solution concentration, shaking time and filtration volume of standard solution are required to be well controlled and selected to obtain more accurate results for TEP with different concentrations.

As an important land-ocean interaction process, submarine groundwater discharge (SGD) is composed of multiple dynamical processes at different scales and plays an important role in the study of coastal ocean geochemical budgets. However, most of the existing studies focus on the quantification of the total groundwater discharge, few studies are about the differentiation and quantification of groundwater discharge processes at different scales (i.e., short-scale SGD and long-scale SGD). As a world-class river, the Huanghe River is highly turbid and heavily regulated by humans. These natural and anthropogenic factors have a significant impact on groundwater discharge processes in the Huanghe River Estuary (HRE). In this study, the distribution patterns of the natural geochemical tracer radium isotopes (²²⁴Ra and ²²³Ra) and other hydrological parameters in the HRE were investigated during four cruises. By solving the mass balance of ²²⁴Ra and ²²³Ra in the HRE, the long-scale SGD flux was quantified as 0.01−0.19 m/d, and the short-scale SGD flux was 0.03−0.04 m/d. The rate of short-scale SGD remained essentially constant among seasons, while the rate of long-scale SGD varied considerably at different periods and showed a synchronous trend with the variation of river discharge. The results of this study are significant for understanding the SGD dynamics in the HRE and the contribution of SGD to the ocean geochemical budgets.

Metallothionein(MT) is a low molecular weight (MW)and heatstable metal-binding prorein.It has been proved that the synthesis of MT in marine animals can be induced by various heavy metals at genetic level.Consequently, high level of MT can be used as sign of heavy metal pollution.
In this article, red scorponifish MT was purified by Sephadex G-75 gel filtration and DEAE-Sepharose CL-6B ion exchange chromatography.The results of amino acids composition analysis showed that the protein contains 35%cysteine (Cys),but no aromatic residues.MW was ahout 6.7 kD, and pI is 4.0 and 4.2,respectively.The antibody was raised in white rabbits by injection by MT conjugated with BSA, and then purified on Protein A beads and labeled with horseradish peroxidase.Quantification of MT by ELISA indicated that the concentration of MT from induced and noninduced fish liver extraction were 450 and 20 μg/cm³,respectively.

The ecosystem-based management of nearshore waters requires integrated assessment of ocean health and scientific guidance on artificial regulations to promote sustainable development. Quantitative approaches were developed in this paper to assess present and near-term ocean health based on ecosystem services. Results of the case study in the Laizhou Bay of China showed that the index score of ocean health was 0.7856 out of 1.0 at present and was expected to range from 0.5551 to 0.8041 in the near-term future depending on different intensities of artificial regulation of negative pressures. Specifically, the results of ocean health at present mainly indicated that cultural services and provisioning services performed essentially perfectly while supporting services and regulating services functioned less well. It can be concluded that this nearshore ecosystem would partially lose supporting and regulating services in the near-term future if the increasing pressures were not wellregulated but that all of these categories of ecosystem services could be slightly improved if the negative pressures were fully controlled. Additionally, it is recommended that publicity and education on ecosystem services especially on cultural services and regulating services should be further strengthened. The analytical process and resulting quantification provide flexible tools to guide future development of regulations so as to facilitate ecosystem-based management in the coastal zone.

The study was to find out the effect of cadmium and mercury on levels of hemolymph glucose, crustacean hyperglycemic hormone (CHH) and hepatopancreas glycogen in the blue swimmer crab Portunus pelagicus. The experiments were performed in both intact and eyestalk ablated crabs. Quantification of CHH was done by the indirect ELISA with the aid of primary anti-Carcinus maenas-CHH antibody. Higher glucose concentration was observed on exposure to 8×10^-6 of cadmium ((825.6±5.42) μg/mL) and 6×10^-6 of mercury ((90.5±6.25) μg/mL) after 48 h and 24 h respectively. Higher level of hemolymph glucose was observed in eyestalk intact crabs on exposure to cadmium and mercury than eyestalk ablated crabs. Decrease in the levels of CHH was observed in both eyestalk intact and ablated crabs on heavy metal exposure. Decline of the hepatopancreas glycogen level was also witnessed with the exposure to heavy metal, which validated its utilization in the production of glucose. Thus this study brings to light, the variations in hemolymph glucose, CHH and hepatopancreas glycogen on heavy metal stress. These carbohydrate metabolites can be used as biomarkers in assessing heavy metal contamination in water bodies.

Edwardsiella tarda is one of the most important emerging pathogens in the global aquaculture industries. As such, an accurate diagnosis and quantitative analytical methods are urgently needed for this bacterium. In this study, primers and a TaqMan probe specific to the conservative sequences of the 16S rRNA gene of E. tarda were designed. The concentration of primers and TaqMan probe were optimized to 200 nmol/L and 120 nmol/L, respectively. The detection sensitivity of the FQPCR assay was determined to be as low as five copies of the target sequence per reaction using the pGEM-16S rDNA recombinant plasmid as a template, which was 100 times more sensitive than conventional PCR. A standard curve by plotting the threshold cycle values (y) against the common logarithmic copies (log₁₀n_c as x; n_c is copy number) of pGEM-16S rDNA was generated. The results of intra-and inter-assay variability tests demonstrate that the established FQ-PCR method was highly reproducible. The assay was specific for E. tarda as it showed that there was no cross-reactivity to eight additional bacterial pathogen strains in aquaculture. Thus, the FQ-PCR assay has the potential for diagnostic purposes and for other applications, especially for the rapid detection and quantification of low-grade E. tarda infections.