Lignin in marine environment and its analysis—A review

Lignin is a group of phenolic polymers which is abundant in the woody tissues of vascular plants,and is essentially absent from all other living organisms.It has therefore been accepted as a tracer for terrestrial organic carbon(TOC) in marine environment since the 1970s.Lignin polymers are not amenable to direct chemical analysis without prior isolation.This review focused on the methods of chemical decomposition,extraction,derivatization and detection of lignin in marine environment.We described and compared several chemical decomposition methods,including nitrobenzene oxidation,alkaline cupric oxide(CuO) oxidation and thermochemolysis,and detection methods such as gas chromatography(GC),gas chromatography-mass spectrometry(GC-MS),high performance liquid chromatography(HPLC) and so on.Possible improvement of lignin analysis and the application prospects of this tracer were also discussed.     

Effect of co-culture of Chinese shrimp (<Emphasis Type="Italic">Fenneropenaeus chinensis</Emphasis>) and sea cucumber (<Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka) on pond environment

Monoculture of sea cucumber (pond S) and polyculture of shrimp with sea cucumber (pond SS) were established to evaluate the effect of shrimp on the environmental conditions of sea cucumber farming pond. Contributions of sediment organic matter (SOM2) resuspended from benthic sediment and the suspended particulate organic matter (SPOM) deposited from the water column to the precipitated organic matter (SOM1) collected with sediment traps were estimated with carbon stable isotope analysis. The results showed that the levels of SPOM and SOM2 in pond SS significantly decreased in comparison with those in pond S at the end of experiment (P < 0.05), indicating that co-culturing shrimp in sea cucumber farming pond could purify the farming water. Carbon stable isotope analysis showed that the proportion of SOM2 in SOM1 in pond SS (84.97% ± 0.38%) was significantly lower than that in pond S (95.20% ± 0.30%) (P < 0.05), suggesting that the resuspension of organic matter from benthic sediment into overlying water was reduced in polyculture pond. In contrast, the proportion of SPOM in SOM1 in pond SS (15.03% ± 0.38%) was significantly higher than that in pond S (4.80% ± 0.30%) (P < 0.05), indicating that the sedimentation of SPOM from water column was enhanced in pond SS owing to the biodeposition effect of shrimp.     

Optimization of ultrasonic extraction and clean-up protocol for the determination of polycyclic aromatic hydrocarbons in marine sediments by high-performance liquid chromatography coupled with fluorescence detection

The procedures of ultrasonic extraction and clean-up were optimized for the determination of polycyclic aromatic hydrocarbons (PAHs) in marine sediments. Samples were ultrasonically extracted, and the extracts were purified with a miniaturized silica gel chromatographic column and analyzed with high performance liquid chromatography (HPLC) with a fluorescence detector. Ultrasonication with methanol-dichloromethane (2:1, v/v) mixture gave higher extraction efficiency than that with dichloromethane. Among the three elution solvents used in clean-up step, dichloromethane-hexane (2:3, v/v) mixture was the most satisfactory. Under the optimized conditions, the recoveries in the range of 54.82% to 94.70% with RSDs of 3.02% to 23.22% for a spiked blank, and in the range of 61.20% to 127.08% with RSDs of 7.61% to 26.93% for a spiked matrix, were obtained for the 15 PAHs studied, while the recoveries for a NIST standard reference SRM 1941b were in the range of 50.79% to 83.78% with RSDs of 5.24% to 21.38%. The detection limits were between 0.75 ng L-1 and 10.99 ng L-1for different PAHs. A sample from the Jiaozhou Bay area was examined to test the established methods.     

Studies on the community structures of meiofauna and marine nematode at six stations in the Southern Yellow Sea,China

1Introduction As part of a comprehensive investigation of ecosys-tem dynamics in the Yellow Sea and East China Sea,astudy has been made of meiofauna,defined here asmetazoans passing a0.5mm sieve but retained by a0.031mm sieve.Meiofauna is an important energeticgroup in benthic small food web due to their smallsize,high abundance and fast turnover rates.Theproduction of meiofauna is equal to or higher than thatof macrofauna in estuaries,shallow waters and deepseas(Gerlach,1971;Platt and Warw…     

Antifouling potential of bacteria isolated from a marine biofilm

Marine microorganisms are a new source of natural antifouling compounds. In this study, two bacterial strains, Kytococcus sedentarius QDG-B506 and Bacillus cereus QDG-B509, were isolated from a marine biofilm and identified. The bacteria fermentation broth could exert inhibitory effects on the growth of Skeletonema costatum and barnacle larvae. A procedure was employed to extract and identify the antifouling compounds. Firstly, a toxicity test was conducted by graduated pH and liquid-liquid extraction to determine the optimal extraction conditions. The best extraction conditions were found to be pH 2 and 100% petroleum ether. The EC50 value of the crude extract of K. sedentarius against the test microalgae was 236.7 ± 14.08 μg mL-1, and that of B. cereus was 290.6 ± 27.11 μg mL-1. Secondly, HLB SPE columns were used to purify the two crude extracts. After purification, the antifouling activities of the two extracts significantly increased: the EC50 of the K. sedentarius extract against the test microalgae was 86.4 ± 3.71 μg mL-1, and that of B. cereus was 92.6 ± 1.47 μg mL-1. These results suggest that the metabolites produced by the two bacterial strains are with high antifouling activities and they should be fatty acid compounds. Lastly, GC-MS was used for the structural elucidation of the compounds. The results show that the antifouling compounds produced by the two bacterial strains are myristic, palmitic and octadecanoic acids.     

TRANSFERABLE PHOSPHORUS IN SEDIMENTS OF THE HUANGHE RIVER ESTUARY ＇S ADJACENT WATERS

Based on a new idea for research on cycling of marine biogenic elements, this study showed that only the leachable form phosphorus in natural grain sizes manne sediments constitutes the transferable phosphorous in the sediments. The transferable phosphorus content in the natural grain sizes surface sediments in the Huanghe River estuary adjacent waters ranges from 58.5-69.8 μg/g, accounting for only 9.1%-11.0% of the total phosphorus content, whereas the leachable form (“transferable” )phosphorus content in the sediment after it was totally ground into powder was found to be 454.8-529.2μg/g, accounting for 73.4%-89.1% of the total phosphorus. Analysis of the correlation between thebill,ass of benthos and the leachable form (“transferable” ) phosphorus showed that most of the leachableform (“transferable”) phosphorus in the totally ground sediment did not participate in the marine biogeo chemical cycling. Furthermore, a synchronous survey on benthos showed that the biornass of meio-and maero-benthos exhibited good positive correlation with the leachable form of phosphorus in the naturalgrain sizes sediment, but peorer correlation with the leachable form (“transferable”) phosphorus in the totally ground sediment, indicating that transferable phosphorus in marine sediment is the leachable form of phosphorus in the natural grain sizes sediments, and is not the previously known leachable form(“transferable”) phosphorus obtained from the totally ground sediment.     

A new method for the estimation of fine-sediment resuspension ratios in estuaries-taking the turbidity maximum zone of the Changjiang(Yangtze) estuary as an example

Based on the principle of conservative matter removal in estuary,a new method is proposed for estimating the ratio of sediment resuspension in estuaries with fine suspended sediments in the turbidity maximum zone(TMZ) of the Changjiang(Yangtze) estuary during 2005.Results show that there was a range of 18.7%±27.9% to 73.9%±22.5% per annum of total suspended particulate matter(SPM),with an average of 49.2%.Nearly half of the particulate matter in the TMZ originates from sediment resuspension.This indicates that sediment resuspension is one of the major mechanisms involved in formation of the TMZ.Compared with traditional method for calculating these ratios in the estuary,this new method evaluates the dynamic variation of SPM content carried by river runoff from the river mouth to the ocean.The new method produced more reliable results than the traditional one and could produce a better estimation of resuspension flux for particulate matter in estuaries.     

Lipid and fatty acid compositions of cod (<Emphasis Type="Italic">Gadus morhua</Emphasis>), haddock (<Emphasis Type="Italic">Melanogrammus aeglefinus</Emphasis>) and halibut (<Emphasis Type="Italic">Hippoglossus hippoglossus</Emphasis>)

This study was conducted to compare lipid and fatty acid composition of cod, haddock and halibut. Three groups of cod (276 g ± 61 g), haddock (538 g ± 83 g) and halibut (3704 g ± 221 g) were maintained with commercial feeds mainly based on fish meal and marine fish oil for 12 weeks prior to sampling. The fatty acid compositions of muscle and liver were determined by GC/FID after derivatization of extracted lipids into fatty acid methyl esters (FAME). Lipids were also fractionated into neutral and polar lipids using Waters silica Sep-Pak?. The phospholipid fraction was further separated by high-performance thin-layer chromatography (HPTLC) and the FAME profile was obtained. Results of the present study showed that cod and haddock were lean fish and their total muscle lipid contents were 0.8% and 0.7%, respectively, with phospholipid constituting 83.6% and 87.5% of the total muscle lipid, respectively. Halibut was a medium-fat fish and its muscle lipid content was 8%, with 84% of the total muscle lipid being neutral lipid. Total liver lipid contents of cod, haddock and halibut were 36.9%, 67.2% and 30.7%, respectively, of which the neutral lipids accounted for the major fraction (88.1%–97.1%). Polyunsaturated fatty acids were the most abundant in cod and haddock muscle neutral lipid. Monounsaturated fatty acid level was the highest in halibut muscle neutral lipid. Fatty acid compositions of phospholipid were relatively constant. In summary, the liver of cod and haddock as lean fish was the main lipid reserve organ, and structural phospholipid is the major lipid form in flesh. However, as a medium-fat fish, halibut stored lipid in both their liver and muscle.     

Microanalysis and preliminary pharmacokinetic studies of a sulfated polysaccharide from <Emphasis Type="Italic">Laminaria japonica</Emphasis>

A rapid, sensitive and reproducible high performance liquid chromatography (HPLC) method with post-column fluorescence derivatization has been developed to determine the amount of low-molecular-weight sulfated polysaccharide (GFS) in vivo. The metabolism of GFS has been shown to fit a two component model following its administration by intravenous injection, and its pharmacokinetic parameters were determined to be as follows: half-time of distribution phase (t _1/2α)=11.24±2.93 min, half-time of elimination phase (t _1/2β)=98.20±25.78 min, maximum concentration (C _max)=110.53 μg/mL and peak time (T _max)=5 min. The pharmacokinetic behavior of GFS was also investigated following intragastric administration. However, the concentration of GFS found in serum was too low for detection, and GFS could only be detected for up to 2 h after intragastric administration (200 mg/kg body weight). Thus, the bioavailability of GFS was low following intragastric administration because of the metabolism of GFS. In conclusion, HPLC with postcolumn derivatization could be used for quantitative microanalysis and pharmacokinetic studies to determine the presence of polysaccharides in the serum following intravenous injection.     

Production of polyhydroxybutyrate by the marine photosynthetic bacterium Rhodovulum sulfidophilum P5

The effects of different NaCl concentrations, nitrogen sources, carbon sources, and carbon to nitrogen molar ratios on biomass accumulation and polyhydroxybutyrate (PHB) production were studied in batch cultures of the marine photosynthetic bacterium Rhodovulum sulfidophilum P5 under aerobic-dark conditions. The results show that the accumulation of PHB in strain P5 is a growth-associated process. Strain P5 had maximum biomass and PHB accumulation at 2%-3% NaCl, suggesting that the bacterium can maintain growth and potentially produce PHB at natural seawater salinity. In the nitrogen source test, the maximum biomass accumulation (8.10±0.09 g/L) and PHB production (1.11±0.13 g/L and 14.62%±2.25% of the cell dry weight) were observed when peptone and ammonium chloride were used as the sole nitrogen source. NH 4 + -N was better for PHB production than other nitrogen sources. In the carbon source test, the maximum biomass concentration (7.65±0.05 g/L) was obtained with malic acid as the sole carbon source, whereas the maximum yield of PHB (5.03±0.18 g/L and 66.93%±1.69% of the cell dry weight) was obtained with sodium pyruvate as the sole carbon source. In the carbon to nitrogen ratios test, sodium pyruvate and ammonium chloride were selected as the carbon and nitrogen sources, respectively. The best carbon to nitrogen molar ratio for biomass accumulation (8.77±0.58 g/L) and PHB production (6.07±0.25 g/L and 69.25%±2.05% of the cell dry weight) was 25. The results provide valuable data on the production of PHB by R. sulfidophilum P5 and further studies are on-going for best cell growth and PHB yield.     

An approach to determination of phenolic compounds in seawater using SPME-GC-MS based on SWCNTs coating

Phenolic compounds have become one kind of the important pollutants of the marine environment. Single-walled Carbon nanotubes, as one-dimensional nano materials, have light weight and perfect hexagonal structure of connections, with many unusual mechanical, chemical and electrical properties. In recent years, with the research of carbon nanotubes and other nano materials, the application prospect is also constantly discussed. In this paper, homemade single-walled carbon nanotubes(SWCNTs) coating was used for establishing an analytical approach to the determination of five kinds of phenolic compounds in seawater using SPME-GC-MS. Optimal conditions: After saturation was conducted with Na Cl, and p H was adjusted to 2.0 with H_2SO_4, the extract was immersed in a water bath at 40 for GC℃-MS determination through 40-min agitating extraction at 500 rmin~(-1) and 3-min desorption at 280℃. The liniearities ranged between 0.01-100 μg L~(-1), and the determination limits ranged between 1.5-10 ngL~(-1). The relative standard deviation(RSD, n = 5) was less than 6.5%. For the phenolic compounds obtained from the spiked recovery test for actual seawater samples, the rates of recovery were 87.5%-101.7%, and the RSDs were less than 8.8%, which met the requirements of determination. Due to its simplicity, high efficiency and low consumption, this approach is suitable for the analysis of trace amounts of phenolic compounds in marine waters.     

Biodiversity of free-living marine nematodes in the southern Yellow Sea,China

Biodiversity patterns of free-living marine nematodes were studied using specific,taxonomic and phylogenetic diversity measures in the southern Yellow Sea,China.The results showed that the average of Shannon-Wiener diversity index(H′) in the study area was 3.17.The higher values were distributed in the east part of Shandong coastal waters and north part of Jiangsu coastal waters,while the lower values were distributed in the southern Yellow Sea Cold Water Mass(YSCWM).The average of taxonomic diversity(Δ) was 62.09 in the study region.The higher values were distributed in the transitional areas between the coastal areas and the southern YSCWM,while the lower values were distributed near the north part of Jiangsu coastal waters and the YSCWM.Results of correlation analysis of species diversity and taxonomic diversity showed that some of the two kinds of diversity index were independent,which suggested that combining the two kinds of diversity indices can reflect the ecological characteristics better.A test for 95% probability funnels of average taxonomic distinctness and variation in taxonomic distinctness suggested that Station 8794(in the YSCWM) was outside of the 95% probability funnels,which may be due to the environmental stress.Results of correlation analysis between marine nematodes biodiversity and environmental variables showed that the sediment characteristics(Md? and Silt-clay fraction) and phaeophorbide a(Pha-a) were the most important factors to determine the biodiversity patterns of marine nematodes.     

Effects of <Emphasis Type="Italic">Urechis unicinctus</Emphasis> juveniles on chemical characteristics of organically contaminated coastal sediment

Biological activities of marine benthos such as burrowing and feeding may change sediment characteristics. We conducted three experiments to examine the potential of using juveniles of a spoon worm Urechis unicinctus to improve the quality of organically contaminated coastal sediment. Sediment samples were collected from a site that was heavily contaminated with organic matter (Seonso) and two sites that were clean (Myo-do, Dolsan-do). Urechis juveniles, obtained by artificial fertilization and cultured in the laboratory, were introduced to the sediment (weight 3 kg, depth 10 cm) at a density of 500 individuals per aquarium (length 50 cm, width 35 cm, height 30 cm) (Experiment 1), or at densities ranging from 100 to 900 individuals per beaker (Experiment 2). To examine how sediment contamination can be modified by the effects of Urechis, 500 individuals (per aquarium) were exposed to the Seonso contaminated sediment that had been mixed with 0–100% clean sand (Experiment 3). Each experiment lasted two months and sediment samples were collected every 15 d to determine the several indexes of sediment quality, which included acid volatile sulfide (AVS), chemical oxygen demand (COD) and total ignition loss (TIL). In Experiment 1, the existence of Urechis did not result in significant changes in quality indexes in the sediments collected from Myo-do, Dolsan-do. However, AVS, COD and TIL of the Seonso sediment all decreased significantly after co-incubation with Urechis juveniles for 30 to 45 d. Experiment 2 showed that a density of at least 300 juveniles per beaker was necessary to significantly reduce all three quality indexes, and the magnitude of reduction was positively correlated with juvenile density. Experiment 3 revealed that Urechis juveniles were effective in reducing the AVS, COD and TIL of the Seonso sediment that had been mixed with 60%, 80%, and 80% of clean sand, respectively. The results of the present study therefore indicated that juveniles of this spoon worm have the potential to be used to improve the quality of organically contaminated sediment in coastal waters.     

Short- and long-term response of phytoplankton to ENSO in Prydz Bay,Antarctica: Evidences from field measurements,remote sensing data and stratigraphic biomarker records

The study provides one of the first lines of evidence showing linkages between Antarctic phytoplankton abundance and composition in response to ENSO, based on historical reconstruction of sediment biomarkers. In addition to sediment biomarkers, field measured and remote sensing data of phytoplankton abundance were also recorded from Prydz Bay, Eastern Antarctica. Com-munity structure of field measured phytoplankton showed significant El Ni？o/La Ni？a-related succession during 1990 to 2002. In general, the number of algae species decreased during El Ni？o and La Ni？a years compared to normal years. Austral summer monthly variation of remotely sensed chlorophyll-a （Chl-a）, particulate organic carbon （POC）, and sea surface temperature （SST） indicated that ENSO impacted the timing of phytoplankton blooms during 2007 to 2011. Phytoplankton blooms （indicated by Chl-a and POC） preceded the increases in SST during El Ni？o years, and lagged behind the SST increases during La Ni？a years. Stratigraphic record of marine sedimentary lipid （brassicasterol, dinosterol and alkenones） biomarkers inferred that the proportions of different algae （diatoms, dinoflagellates and haptophytes） changed significantly between El Ni？o and La Ni？a events. The relative proportion of diatoms increased, with that of dinoflagellates being decreased during El Ni？o years, while it was reversed during La Ni？a years.     

澳门附近海域沉积物污染状况及特征分析

掌握澳门附近海域沉积物的污染状况及特征,对于合理安排澳门海洋海岸工程疏浚泥的海洋倾倒,防止造成海洋污染有十分重要的意义。根据1997—1999年连续三年对澳门附近主要区域的海洋沉积物采样测定结果,通过分析认为澳门附近海域沉积物污染具有下列特征:1)沉积物中重金属和油类含量普遍较高,除了总Hg外均高于珠江口海区的其他港口;2)沉积物中污染物的分布受人类活动的影响明显,澳门内港受船只排污、码头作业、陆地工业和生活污水排放的影响,是澳门附近海域沉积物污染最为严重区域;3)受水质交换的影响明显,澳门外港、路环岛东北等区域受珠江径流影响水质交换较快,沉积物中污染物的含量较低,而内港由于水质交换较差,沉积物中污染物的含量相对较高;4)与沉积物的成分组成(吸附)有关,大部分污染物与沉积物中粉砂有较好的相关性,表明粉砂对污染物质有较广泛性的吸附能力,尤其是Cu、Zn、Cd、油类、有机质。     

ANALYSIS AND DISTRIBUTION FEATURES OF BENZOTHIOPHENE IN SEDIMENTS OF NANSHA ISLANDS SEA AREA

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Geochemical analysis of marine sediments using fused glass disc by X-ray fluorescence spectrometry

A method was developed for content determination of Na, Mg, A1, Si, P,S, C1, K, Ca, Ti, V, Cr, Mn, Fe, Ni, Cu, Zn, Ga, Nb, Zr, Y, Sr, Rb, Ba, La and Ce etc. covering 26 major, minor, and trace elements in marine sediment samples using fused glass disc by X-ray Fluorescence spectrometry. Calibration was made using marine sediment certified reference materials and the synthetic standard samples prepared by mixing several marine sediments with stream sediment and carbonate standard samples in different proportions. The matrix effect was corrected using theoretical alpha coefficients, experience coefficients and the scattered radiation as the internal standard （for the trace elements）. The accuracy of the method was evaluated by analysis of certified reference materials GBW07314, GBW07334 and GSMS6. The results are in good agreement with the certified values of the standards with RSD less than 2.60%, except for Y, Cr, Ga, Ce, La, Nb, Rb, and V with RSD less than 9.0% （n=12）.     

Response to selection for fast growth in the second generation of Pacific oyster (Crassostrea gigas)

Mass selection for fast growth was conducted in three Pacific oyster (Crassostrea gigas) stocks from China, Japan and Korea using previously established lines (CS1, JS1, and KS1). To determine whether continuous progress can be achieved by selection for growth, the progeny of three second-generation Pacific oyster lines was selected for shell height and evaluated via a 400-day farming experiment. When harvested at the end of the experiment, the selected crosses of CS2, JS2, and KS2 lines grew by 9.2%, 10.2% and 9.6% larger than the control crosses, respectively. During grow-out stage, the genetic gain of three selected lines was (10.2 ± 1.4)%, (10.4 ± 0.3)%, and (8.4 ± 1.6)%, respectively; and the corresponding realized heritability was 0.457 ± 0.143, 0.312 ± 0.071 and 0.332 ± 0.009, respectively. These results indicated that the selection for fast growth achieved steady progress in the second generation of oyster. Our work provides supportive evidence for the continuity of the Pacific oyster selective breeding program.     

Size-fractionated Chlorophyll <Emphasis Type="Italic">a</Emphasis> biomass in the northern South China Sea in summer 2014

Spatial distribution of phaeopigment and size-fractionated chlorophyll a (Chl a) concentrations were examined in relation to hydrographic conditions in the northern South China Sea (NSCS) during a survey from 20 August to 12 September, 2014. The total Chl a concentration varied from 0.006 to 1.488 µg/L with a mean value of 0.259±0.247 (mean±standard deviation) µg/L. Chl a concentration was generally higher in shallow water (<200 m) than in deep water (>200 m), with mean values of 0.364±0.311 µg/L and 0.206±0.192 µg/L respectively. Vertically, the maximum total Chl a concentration appeared at depths of 30–50 m and gradually decreased below 100 m. The size-fractionated Chl a concentrations of grid stations and time-series stations (SEATS and J4) were determined, with values of pico- (0.7–2 µm), nano- (2–20 µm) and micro-plankton (20–200 µm) ranging from 0.001–0.287 (0.093±0.071 µg/L), 0.004–1.149 (0.148±0.192 µg/L) and 0.001–0.208 (0.023±0.036 µg/L), respectively. Phaeopigment concentrations were determined at specific depths at ten stations, except for at station A9, and varied from 0.007 to 0.572 (0.127±0.164) µg/L. Nano-and pico-plankton were the major contributors to total phytoplankton biomass, accounting for 50.99%±15.01% and 39.30%±15.41%, respectively, whereas microplankton only accounted for 9.39%±8.66%. The results indicate that the contributions of microplankton to total Chl a biomass were less important than picoplankton or nanoplankton in the surveyed NSCS. Different sized-Chl a had similar spatial patterns, with peak values all observed in subsurface waters (30–50 m). The summer monsoon, Kuroshio waters, Zhujiang (Pearl) River plume, and hydrological conditions are speculated to be the factors controlling the abundance and spatial heterogeneity of Chl a biomass in the NSCS.     

THE EVOLUTION OF STENOLS AND STANOLS IN MARINE SEDIMENT CORE FROM BEIBU BAY, THE SOUTH CHINA SEA

Ten stenols and stanols were identified by TLC , GC and GC-MS-DS in a marine sediment core from Beibu Bay , the South China Sea .C27, C28 and C29 A5 stenols and 5a- stanols mainly come from zoopiankton, algae and phytoplankton in marine environment. C29 24-ethylcholest- 5 - en- 3B- ol is thought to indicate input of terrigenous organic matter . The increase of C27 . C28 and C29 5a-stanol /5 stenol ratios with sediment depth shows the evolution of stenols and stanols in the marine sediment core .