Tracing dissolved organic matter in aquatic environments using a new approach to fluorescence spectroscopy

Dissolved organic matter (DOM) is a complex and poorly understood mixture of organic polymers that plays an influential role in aquatic ecosystems. In this study we have successfully characterised the fluorescent fraction of DOM in the catchment of a Danish estuary using fluorescence excitation–emission spectroscopy and parallel factor analysis (PARAFAC). PARAFAC aids the characterisation of fluorescent DOM by decomposing the fluorescence matrices into different independent fluorescent components. The results reveal that at least five different fluorescent DOM fractions present (in significant amounts) in the catchment and that the relative composition is dependent on the source (e.g. agricultural runoff, forest soil, aquatic production). Four different allochthonous fluorescent groups and one autochthonous fluorescent group were identified. The ability to trace the different fractions of the DOM pool using this relatively cheap and fast technique represents a significant advance within the fields of aquatic ecology and chemistry, and will prove to be useful for catchment management.     

Photobleaching of fluorescence and the organic carbon concentration in a coastal environment

In order to investigate the photobleaching potential of estuarine waters from different depths and redox conditions and with varying degree of biological activity, filtered, unfiltered and chloroform-poisoned water samples from the Baltic Sea were exposed to ambient sunlight. Fluorescence, at excitation 350 nm and emission 450 nm, was used as an indication of humic substance concentration. Fluorescence and organic carbon concentration were measured at regular time intervals during light exposure. We found that the decrease in humic substance fluorescence can be fitted to an exponential decay function. The fluorescence half-lives were within the range 0.4 – 4.6 days in different water masses, with fluorescence decreasing to between 20% and 60% of initial concentration, respectively. Results from the curve fitting procedure indicate a rest concentration of humic substance fluorescence, similar among the sampled sites, that is resistant to further photochemical degradation. The largest relative decreases in fluorescence were found in deep waters, but samples from deep waters also had a higher fluorescence rest concentration than samples from surface waters. Biological activity was reduced by filtering the samples through 0.2μm pore size filters or adding chloroform. No statistically significant differences were found after 3 days of irradiation between samples with and without treatment to reduce biological activity. The highest initial fluorescence values and the largest fluorescence decrease were found in the anoxic waters of the Gotland Deep. The organic carbon concentrations decreased 3–7% at all stations. The shortest half-life of humic substance, and the largest decrease in organic carbon concentrations, were found in samples from the northern basins of the Baltic Sea.     

Relative molar mass distributions of chromophoric colloidal organic matter in coastal seawater determined by Flow Field-Flow Fractionation with UV absorbance and fluorescence detection

A new method for the characterization of chromophoric colloidal organic matter in seawater has been applied to samples from the Baltic Sea, Kattegatt and Skagerrak seas. Size fractionation of the sample by Flow Field-Flow Fractionation and measurement of the fluorescent and UV absorbing properties of the individual size fractions result in a relative molar mass distribution (RMM) of the optical properties. The RMM distributions have been used to estimate number and weight average relative molar masses, and polydispersity indices. At least two sources of coloured organic matter were identified from the ratio of fluorescence to UV: the Baltic surface water and the Skagerrak deep water. The dominating processes were mixing and dilution, but processes such as photo bleaching of fluorescence are also believed to be important. The RMM distribution derived from UV detection (1150–1300 Dalton) increased with increasing salinity while that derived for fluorescence (1500–1250 Dalton) decreased with increasing salinity. The specific UV absorbance taken as a proxy of the aromaticity of the chromophoric organic material showed decreasing trend with both increasing salinity and increasing UV derived weight average relative molar mass. Increasing polydispersity of the colloidal material was also observed as a function of salinity.     

A method to measure the isotopic (C) composition of dissolved organic carbon using a high temperature combustion instrument

The stable isotopes of dissolved organic carbon (DOC) are a powerful tool for distinguishing sources and inputs of organic matter in aquatic systems. While several methods exist to perform these analyses, no labs routinely utilize a high temperature combustion (HTC) instrument. Advantages of HTC instruments include rapid analysis, small sample volumes and minimal sample preparation, making them the favored devices for most routine oceanic DOC concentration measurements. We developed a stable carbon DOC method based around an HTC system. This method has the benefit of a simple setup, requiring neither vacuum nor high pressures. The main drawback of the method is a significant blank, requiring careful accounting of all blank sources for accurate isotopic and concentration values. We present here a series of experiments to determine the magnitude, source and isotopic composition of the HTC blank. Over time, the blank is very stable at  20 ng of carbon with a δ¹³C of − 18.1‰ vs. VPDB. The similarity of the isotopic composition of the blank and seawater samples makes corrections relatively minor. The precision of the method was determined by oxidizing organic standards with a wide isotopic and concentration range (− 9‰ to − 39‰; 18 μM to 124 μM). Analysis of seawater samples demonstrates the accuracy for low concentration, high salinity samples. The overall error on the measurement is approximately ± 0.8‰.     

错流超滤技术在海水胶体态铀、钍、镭同位素和有机碳研究中的应用

建立了由预过滤装置、蠕动泵、中空纤维超滤膜(AmiconH10P10-20,标称截留分子量10KDa)和连接管组成的错流超滤系统,利用荧光标记的40KDa葡聚糖和已知放射性活度的234Th示踪剂评估了超滤膜的截留和吸附性质,探讨了234Th在超滤过程中的渗透行为,考查了该系统用于实际海水样品时铀、钍、镭同位素和有机碳的质量平衡状况.结果表明,10Kda中空纤维超滤膜对40Kda葡聚糖具有良好的截留效率(85%),而吸附损失率为18%.铀、钍、镭同位素和有机碳在超滤过程中均达到极佳的质量平衡,回收率R=95%～98%,优于大多数文献报道的值.234Th在超滤过程中的渗透行为可以很好地用渗透模型加以描述.研究组分胶体态含量占“溶解”态含量的份额大小顺序如下:钍同位素、有机碳、镭同位素约等于铀同位素,这与钍为强颗粒活性元素、铀和镭为水溶性元素的地球化学性质相吻合.     

Characterization of chromophoric dissolved organic matter (CDOM) in the Baltic Sea by excitation emission matrix fluorescence spectroscopy

Chromophoric dissolved organic matter (CDOM) is the major light absorber in the Baltic Sea. In this study, excitation emission matrix (EEM) fluorescence spectra and UV–visible absorption spectra of CDOM are reported as a function of salinity. Samples from different locations and over different seasons were collected during four cruises in 2002 and 2003 in the Baltic Sea in both Pomeranian Bay and the Gulf of Gdansk. Absorption by CDOM decreased with increased distance from the riverine source and reached a relatively stable absorption background in the open sea. Regression analysis showed that fluorescence intensity was linearly related to absorption by CDOM at 375 nm and a_CDOM(375) absorption coefficients were inversely related to salinity. Analysis of CDOM-EEM spectra indicated that a change in composition of CDOM occurred along the salinity gradient in the Baltic Sea. Analysis of percent contribution of respective fluorophore groups to the total intensity of EEM spectra indicated that the fluorescence peaks associated with terrestrial humic components of the CDOM and total integrated fluorescence decreased with decreasing CDOM absorption. In contrast, the protein-like fraction of CDOM decreased to a lesser degree than the others. Analysis of the percent contribution of fluorescence peak intensities to the total fluorescence along the salinity gradient showed that the contribution of protein-like fluorophores increased from 2.6% to 5.1% in the high-salinity region of the transect. Fluorescence and absorption changes observed in the Baltic Sea were similar to those observed in similar transects that have been sampled elsewhere, e.g. in European estuaries, Gulf of Mexico, Mid-Atlantic Bight and the Cape Fear River plume in the South Atlantic Bight, although the changes in the Baltic Sea occurred over a much smaller salinity gradient.     

Characterization of dissolved organic matter fluorescence in the South Atlantic Bight with use of PARAFAC model: Interannual variability

Systematic water sampling for characterization of chromophoric dissolved organic matter (CDOM) in the coastal South Atlantic Bight, was conducted as part of the long term Coastal Ocean Research and Monitoring Program (CORMP). Water samples were collected during a 3.5 year period, from October 2001 until March 2005, in the vicinity of the Cape Fear River (CFR) outlet and in adjacent Onslow Bay (OB). During this study there were two divergent hydrological and meteorological conditions in the CFR drainage area: a severe drought in 2002, followed by the very wet year of 2003. CDOM was characterized optically by the absorption coefficient at 350 nm, the spectral slope coefficient (S), and by Excitation Emission Matrix (EEM) fluorescence. Parallel Factor Analysis (PARAFAC) was used to assess CDOM composition from EEM spectra and six components were identified: three terrestrial humic-like components, one marine humic-like component and two protein-like components. Terrestrial humic-like components contributed most to dissolved organic matter (DOM) fluorescence in the low salinity plume of the CFR. The contribution of terrestrial humic-like components to DOM fluorescence in OB was much smaller than in the CFR plume area. Protein-like components contributed significantly to DOM fluorescence in the coastal ocean of OB and they dominated DOM fluorescence in the Gulf Stream waters. Hydrological conditions during the observation period significantly impacted both concentration and composition of CDOM found in the estuary and coastal ocean. In the CFR plume, there was an order of magnitude difference in CDOM absorption and fluorescence intensity between samples collected during the drought compared to the wet period. During the drought, CDOM in the CFR plume was composed of equal proportions of terrestrial humic-like components (ca. 60% of the total fluorescence intensity) with a significant contribution of proteinaceous substances (ca. 20% of the total fluorescence). During high river flow, CDOM was composed mostly of humic substances (nearly 75% of total fluorescence) with minor contributions by proteinaceous substances. The impact of changes in fresh water discharge patterns on CDOM concentration and composition was also observed in OB, though to a lesser degree.     

Application of fluorescence spectroscopic techniques and probes to the detection of biopolymer degradation in natural environments

The activities and substrate specificities of extracellular enzymes in natural systems are not well understood, despite their critical role in microbial remineralization of organic carbon. These enzymes initiate organic carbon degradation by selectively hydrolyzing high molecular weight substrates to lower molecular weight products which can be transported into cells. A set of single- and dual-labeled fluorescent polysaccharides was synthesized and characterized to explore a variety of approaches for measuring enzymatic hydrolysis of biopolymers via photophysical techniques, focusing particularly on rapid and robust optical techniques which are amenable to field measurements in remote locales. A shotgun-labeling approach yielded dual-labeled probes that exhibited substantial donor fluorophore quenching. The photophysical response of these probes to hydrolysis via purified enzymes was investigated in the lab, and fluorescence polarization proved to be a rapid and reliable technique for monitoring probe hydrolysis. Initial field results were also obtained from hydrolysis experiments in sediment porewaters. Because polarization measurements are rapid and simple, this approach could be used to follow the extracellular enzymatic hydrolysis of a wide range of biopolymers which fuel microbial metabolism.     

Optical properties of low molecular weight and colloidal organic matter: Application of the ultrafiltration permeation model to DOM absorption and fluorescence

Cross-flow ultrafiltration (CFF) is often used to obtain separation and concentration of colloids from bulk natural water samples. Application of the ultrafiltration permeation model allows the quantitative determination of the low molecular weight material (LMW, < 1 kDa) and colloids in bulk dissolved organic matter (DOM) from measurements of time series permeate samples obtained from CFF. Detailed analysis of a Yukon River water sample shows that DOM absorption coefficient and fluorescence follow the permeation model and that the complex spectral optical properties of LMW DOM can be reconstructed from CFF data. A combination of measured and modeled data indicates that the LMW contribution to bulk DOM optical properties obtained from CFF can be grossly underestimated by the use of a low concentration factor (CF, the ratio of initial sample volume to retentate volume). Even at a relatively high CF of 19, optical properties of LMW DOM calculated from measurements of the retentate or integrated permeate would underestimate true values by 5–36%. In the Yukon River sample, LMW dissolved organic carbon represented 26% of the bulk concentration, but only 3–14% of the colored DOM was in the LMW fraction while 31–33% of bulk DOM florescence was due to LMW DOM. The contrasting optical properties of LMW and colloidal DOM support the concept that analysis of bulk DOM absorption and fluorescence properties reveals information about DOM molecular weight.     

Modulation of the particulate organic carbon flux to the ocean by a macrotidal estuary: Evidence from measurements of carbon isotopes in organic matter from the Gironde system

The isotopic composition of the organic carbon of the suspended particulate matter in the Gironde estuary and the Biscay shelf has been measured on a seasonal basis from 1977 to 1982. The δ¹³C values show a progressive change along the estuary and permit an estimate of the proportion of terrestrial carbon in each sample. It is estimated from these data that up to 80% of the continental POC is mineralized in the estuary and 3–16% of the riverborne flux is exported to the shelf.     

Fluxes of organic carbon in a fjord on the west coast of Ireland

The supply of particulate organic carbon (POC) via freshwater influx, and its exchange with the open sea, were estimated for a fjord-like coastal inlet (Killary Harbour, Ireland) during 1981. Biological production and destruction of organic matter within the inlet were also studied. POC levels in rivers feeding the system were correlated with rainfall. C : N ratios suggested an increased proportion of POC of terrestrial origin at stations influenced by freshwater inflow, and throughout the fjord in the non-productive season.A tentative annual carbon budget is presented.     

Primary production and fluxes of organic carbon to the seabed in the Russian Arctic seas as a response to the recent warming

The primary production and fluxes of organic matter to the seabed and their variations were estimated in the Norwegian, Greenland, Barents, Kara, Laptev, East Siberian, and Chukchi seas in 2003–2008 on the basis of satellite and field data. When counting the open water area with the assumptions made for the assessment of the primary production in the regions hidden under clouds, the reliable trends of its variability (increasing) were revealed only in the Greenland, Barents, and Kara seas.     

Use of the Comet assay to assess DNA damage in hemocytes and digestive gland cells of mussels and clams exposed to water contaminated with petroleum hydrocarbons

Oil spills can result in the deposition of large quantities of petroleum hydrocarbons into intertidal and shallow waters seriously impacting bivalve populations. Petroleum hydrocarbons are enriched in polycyclic aromatic hydrocarbons (PAH) and PAH analogs many of which may have potential to damage DNA. The Comet assay is useful for assessing DNA damage and has been used to a limited degree with aquatic organisms, but mostly with studies in vitro. We have carried out studies with the Comet assay to assess the DNA damaging potential of complex mixtures of petroleum hydrocarbons for bivalves. Experiments were carried out with mussels (Mytilus edulis) and clams (Mya arenaria) with dispersions and water soluble fractions of an Arabian crude oil which was also chemically characterized in detail by GC-MS. Pilot studies were first conducted to evaluate test performance and reproducibility. An interindividual coefficient of variation ranging from 17 to 30% was established for the assay with hemocytes and digestive gland cells of both species. Exposure to hydrocarbon fractions had no significant impact on clams. However, an increase in DNA damage was observed at P < 0.1 with digestive gland cells of mussels exposed to aqueous fractions of a light crude oil. These studies have demonstrated a potential for DNA damage in bivalves exposed to oil spills in inshore waters as well as potential for interspecies sensitivity.     

A mechanistic study of the high-temperature oxidation of organic matter in a carbon analyzer

This study examines the mechanism of oxidation of dissolved organic matter (DOM) in a high-temperature combustion (HTC) carbon analyzer. The HTC analyzer, which is commonly used to measure the concentration of dissolved organic carbon (DOC) in seawater, is still largely empirical in its operation, and little information is yet available on the chemical and physical mechanisms responsible for oxidation of DOM. To examine the role of water, which has been hypothesized to be a source of reactive oxidants in HTC analyzers, several experiments were conducted using gaseous hexane samples and several wet/dry carrier gases. Because the quantity of O₂(g) needed to oxidize an injected sample of DOM is quite small, a substantial effort has been made to exclude O₂(g) from the combustion furnace of the HTC instrument. In this modified, “air-tight” instrument, the efficiency of conversion of hexane into CO₂ was greatest in dry O₂ and slightly lower in moist O₂. Hexane was only slightly oxidized in dry N₂, but it was largely converted into CO₂ when moist N₂ was used as the carrier gas. These experiments confirm that water provides reactive oxidizing species (perhaps hydroxyl radical) that rapidly convert hexane into CO₂ in the combustion tube of a carbon analyzer. Additional experiments with aqueous solutions of potassium hydrogen phthalate and a variety of carrier gases support this basic hypothesis.     

Stable carbon isotope distribution of particulate organic matter in the ocean: a model study

The stable carbon isotopic composition of particulate organic matter in the ocean, δ¹³C_POC, shows characteristic spatial variations with high values in low latitudes and low values in high latitudes. The lowest δ¹³C_POC values (−32‰ to −35‰) have been reported in the Southern Ocean, whereas in arctic and subarctic regions δ¹³C_POC values do not drop below −27‰. This interhemispheric asymmetry is still unexplained. Global gradients in δ¹³C_POC are much greater than in δ¹³C_DIC, suggesting that variations in isotopic fractionation during organic matter production are primarily responsible for the observed range in δ¹³C_POC. Understanding the factors that control isotope variability is a prerequisite when applying δ¹³C_POC to the study of marine carbon biogeochemistry. The present model study attempts to reproduce the δ¹³C_POC distribution pattern in the ocean. The three-dimensional (3D) Hamburg Model of the Oceanic Carbon Cycle version 3.1 (HAMOCC3.1) was combined with two different parametrizations of the biological fractionation of stable carbon isotopes. In the first parametrization, it is assumed that the isotopic fractionation between CO₂ in seawater and the organic material produced by algae, _P, is a function of the ambient CO₂ concentration. The two parameters of this function are derived from observations and are not based on an assumption of any specific mechanism. Thus, this parametrization is purely empirical. The second parametrization is based on fractionation models for microalgae. It is supported by several laboratory experiments. Here the fractionation, _P, depends on the CO₂ concentration in seawater and on the (instantaneous) growth rates, μ_i, of the phytoplankton. In the Atlantic Ocean, where most field data are available, both parametrizations reproduce the latitudinal variability of the mean δ¹³C_POC distribution. The interhemispheric asymmetry of δ¹³C_POC can mostly be attributed to the interhemispheric asymmetry of CO₂ concentration in the water. However, the strong seasonal variations of δ¹³C_POC as reported by several authors, can only be explained by a growth rate-dependent fractionation, which reflects variations in the cellular carbon demand.     

Design and application of a semi-automated filtration system to study the distribution of particulate organic carbon in the water column of a coastal upwelling system

The processes controlling the production, transport and ultimate fate of particulate organic carbon (POC) in marine systems play a major role in the global carbon cycle. Direct measurements of POC concentrations in oceanic waters have been used extensively to investigate these processes, but because of the time-intensive effort associated with manual collection and filtration of water samples, this approach often results in limited temporal or spatial coverage. To increase the frequency of POC measurements, we designed and built a semi-automated filtration system (SAFS) that can be linked to a ship's underway surface water flow system and/or a towed profiling vehicle with pumping capabilities. Our results show that, compared to manual filtration, SAFS provides the ability to easily perform filtrations at high enough frequency to measure POC concentrations at spatial and temporal resolutions that are closer to those possible with in situ sensors. When combined with optical measurements, SAFS allows for the characterization of several distinct pools of particles within different geographical and depth regions of the water column off the Oregon Coast. These initial applications yielded data that provide considerable insight into the sources and character of particles in the water column and prove that this approach can be used to gain valuable information on the biogeochemistry of this and other similar ocean margins.     

The cycling and oxidation pathways of organic carbon in a shallow estuary along the Texas Gulf Coast

The cycling and oxidation pathways of organic carbon were investigated at a single shallow water estuarine site in Trinity Bay, Texas, the uppermost lobe of Galveston Bay, during November 2000. Radio-isotopes were used to estimate sediment mixing and accumulation rates, and benthic chamber and pore water measurements were used to determine sediment-water exchange fluxes of oxygen, nutrients and metals, and infer carbon oxidation rates. Using ⁷Be and ²³⁴Th_XS, the sediment-mixing coefficient (D_b) was 4.3 ± 1.8 cm² y⁻¹, a value that lies at the lower limit for marine environments, indicating that mixing was not important in these sediments at this time. Sediment accumulation rates (S_a), estimated using ¹³⁷Cs and ²¹⁰Pb_XS, were 0.16 ± 0.02 g cm⁻² y⁻¹. The supply rate of organic carbon to the sediment-water interface was 30 ± 3.9 mmol C m⁻² d⁻¹, of which ∼10% or 2.9 ± 0.44 mmol C m⁻² d⁻¹was lost from the system through burial below the 1-cm thick surface mixed layer. Measured fluxes of O₂ were 26 ± 3.8 mmol m⁻² d⁻¹ and equated to a carbon oxidation rate of 20 ± 3.3 mmol C m⁻² d⁻¹, which is an upper limit due to the potential for oxidation of additional reduced species. Using organic carbon gradients in the surface mixed layer, carbon oxidation was estimated at 2.6 ± 1.1 mmol C m⁻² d⁻¹. Independent estimates made using pore water concentration gradients of ammonium and C:N stoichiometry, equaled 2.8 ± 0.46 mmol C m⁻² d⁻¹. The flux of DOC out of the sediments (DOC_efflux) was 5.6 ± 1.3 mmol C m⁻² d⁻¹. In general, while mass balance was achieved indicating the sediments were at steady state during this time, changes in environmental conditions within the bay and the surrounding area, mean this conclusion might not always hold. These results show that the majority of carbon oxidation occurred at the sediment-water interface, via O₂ reduction. This likely results from the high frequency of sediment resuspension events combined with the shallow sediment mixing zone, leaving anaerobic oxidants responsible for only ∼10–15% of the carbon oxidized in these sediments.     

Size distribution of colloidal trace metals and organic carbon during a coastal bloom in the Baltic Sea

The physico-chemical speciation of organic carbon and selected metals was measured during a coastal bloom in Ekhagen Bay, Baltic Sea, using ultrafiltration.One important objective with the study was to see if any depletion of trace metals could be measured in the directly bioavailable fraction (<1000 Da, the soluble low molecular weight fraction, LMW) during a plankton bloom. Filters with five different cut-offs were used (1 kD (1000 Da), 5 kD, 10 kD, 100 kD and 0.22 μm) in order to delineate the size distribution of colloidal organic carbon (COC) and trace metals.During the bloom in May, LMW Al, Co, Cu, Mn and Ni concentrations decreased although the colloidal and particulate concentrations were relatively high. Data show that desorption of colloidal and particulate bound trace metals to the LMW fraction was slower than the process depleting the LMW fraction.Estimates of the maximum active uptake of Cu, Ni and Mn by the phytoplankton, and the loss of non-bioactive Al from the LMW fraction, indicate that processes other than active uptake by phytoplankton must contribute to the observed depletion of trace metals in the LMW fraction. Hence, in order to estimate the bioavailable pool of trace metals for plankton during bloom conditions, these other processes must be understood and quantified.Transparent Exopolymeric Particles (TEP, reflecting sugar-rich phytoplankton exudates) increased around eight times during the plankton bloom. We hypothesize that the formation of TEP is a process that might be important for the transfer of trace metals from the LMW to the particulate fraction during the phytoplankton bloom, but the significance of TEP for this depletion in Baltic Sea surface water remains to be shown.     

Vertical distribution of dissolved organic carbon (DOC) in the Western Mediterranean Sea in relation to the hydrological characteristics

Vertical profiles of dissolved organic carbon (DOC) from eight hydrological stations in the Tyrrhenian Sea, Sardinia Channel and Algerian Sea, are reported. DOC exhibits concentrations ranging from 58 to 88 μM in surface water, 43–57 μM in the intermediate layer and 49–63 μM in deep waters. The assessment of the hydrological characteristics allows different water masses in the study area to be identified; moreover, different hydrological processes are observed in the Tyrrhenian and Algerian basins. DOC exhibits different values in the different water masses. The lowest DOC concentrations (43–46 μM) were found in the Tyrrhenian Levantine Intermediate Water (LIW). Correlations between DOC and apparent oxygen utilization (AOU), investigated within each water mass, exhibit different behaviors in the intermediate and deep waters, suggesting the occurrence of different processes of oxygen consumption in the different water masses.     

北冰洋楚科奇海陆架到陆坡表层沉积物有机碳载荷的变化

沉积物单位表面积上吸附的有机碳被广泛用于示踪有机碳载荷的变化。本文研究了北冰洋典型边缘海——楚科奇海表层沉积物的有机碳载荷。研究发现陆架区沉积物的有机碳载荷高于陆坡区。相比于已报道的东西伯利亚海和马更些河,楚科奇海陆坡区沉积物的有机碳载荷也较低。这种有机碳载荷的变化可能和陆坡区的初级生产力较低,以及沉积物在传输过程中经历的氧化降解有关。沉积物的有机碳含量和比表面积呈线性相关,在有机碳轴上有正截距,表明一部分有机碳来自于岩石的贡献。此外,陆架区低有机碳载荷的沉积物含有的岩石有机碳更高。本研究的数据有助于深刻理解楚科奇海区域的碳循环问题。