A new method for the recovery of ammonium from natural waters for measurement of 15N composition in isotope dilution experiments

Efforts to measure the rate of nitrogen recycling in natural waters have depended on estimates of ammonium production using the ¹⁵N isotope dilution method. This technique requires the removal of dissolved ammonium from the water sample for analysis of the ¹⁵N content. Steam distillation or micro-diffusion methods have been most commonly used, but these techniques are labor intensive and time consuming. We have attempted to simplify the measurement of the planktonic regeneration of ammonium by presenting a mercury precipitation technique which is simple, practical, and easily used at sea.     

天然海水硝酸盐氮同位素组成的蒸馏法测定

建立了测定天然海水中硝酸盐氮同位素组成的蒸馏法,该方法主要是在碱性条件下利用戴氏合金将海水中的硝酸盐还原为氨,后利用稀盐酸吸收生产的氨,将得到的氨吸收液浓缩后干燥结晶,利用同位素比值质谱仪测定所得晶体的氮同位素组成。研究中开展了戴氏合金添加量及氨吸收溶液在不同条件下干燥结晶对氮同位素测值的影响研究。结果表明,戴氏合金添加量为3.0 g及60 ℃下直接干燥结晶为最佳的实验条件。所建立的氨蒸馏法氮空白值仅为(0.90±0.19) μmol,低于此前文献报道的氮空白值;氮同位素组成(δ15N)空白值为(-14.7±4.1)×10-3。运用所建立的氨蒸馏法实测得到的硝酸盐δ15N值与氨扩散法、硝酸盐直接测定法得到的数值非常吻合,进一步证明所建立氨蒸馏法的可靠性。改进后的氨蒸馏法适用于硝酸盐浓度在2~50 μmol/dm3内的天然海水硝酸盐氮同位素组成的测定,方法的标准偏差为±0.3×10-3。     

Growth of marine bacteria and ammonium regeneration from substrates in different C: N ratios

Natural assemblages of marine bacteria were chosen in a batch culture experiments. The impact of varying nitrogen substrate concentrations and the substrate C:N ratios (C:N_S) on the bacterial C:N ratio (C:N_B), the bacterial growth efficiency (BGE) and ammonium regeneration was mainly examined. The C:N_S ratios varied from 5:1 (carbon limitation) to 40:1 (nitrogen limitation) with varying combinations of glucose and NO₃^-. The C:N_B ratio had positive relationship with the C:N_S ratio (r=0.93, n=8), whose value was 3.77 when the C:N_S ratio was 5:1 but increased to 6.47 when the C:NS ratio was 40:1. These results indicate that the C:N_B ratio is a potential diagnostic tool for determining the bacterial growth in natural waters controlled by either, carbon or nitrogen. BGE decreased with the declining nitrate concentration and negatively related to C:N_s (r=-0.51, n=8). The average value of BGE was 0.20. This value was a little lower than other reports, which could be induced by the nitrogen source used in our experiments. Finally, regeneration time of ammonium delayed with the increasing C:N_S ratio, which indicates that there were different metabolism mechanisms when bacterial growth was limited by carbon source and nitrogen source.     

Recovery of ammonium nitrogen by solvent extraction for the determination of relative 15N abundance in regeneration experiments

A new sample preparation method for ¹⁵N tracer measurement of ammonium regeneration in seawater is described. Ammonium nitrogen is incorporated into indophenol, extracted into dichloromethane and concentrated by evaporation. The isotopic ratio of the indophenol nitrogen is determined by emission spectroscopy. A small amount (0.2 μmol) of unlabeled nitrogen is introduced by reagents used during sample preparation. For samples of equal nitrogen content, the coefficient of variation is constant and the precision of the isotopic ratio determination is inversely related to the relative ¹⁵N abundance. Differences of 0.19 atom% ¹⁵N can be measured in 5.0 μM NH₄⁺ samples of 5.0 atom% ¹⁵N (SD = 0.07, n = 3, p = 0.05). Interface from forms of dissolved organic nitrogen prevalent in marine environments are negligible.     

Practical considerations for the segmented-flow analysis of nitrate and ammonium in seawater and the avoidance of matrix effects

In this study we describe measures taken in our laboratory to improve the long-term precision of nitrate and ammonia analysis in seawater using a microflow segmented-flow analyzer. To improve the nitrate reduction efficiency using a flow-through open tube cadmium reactor (OTCR), we compared alternative buffer formulations and regeneration procedures for an OTCR. We improved long-term stability for nitrate with a modified flow scheme and color reagent formulation and for ammonia by isolating samples from the ambient air and purifying the air used for bubble segmentation. We demonstrate the importance of taking into consideration the residual nutrient content of the artificial seawater used for the preparation of calibration standards. We describe how an operating procedure to eliminate errors from that source as well as from the refractive index of the matrix itself can be modified to include the minimization of dynamic refractive index effects resulting from differences between the matrix of the samples, the calibrants, and the wash solution. We compare the data for long-term measurements of certified reference material under two different conditions, using ultrapure water (UPW) and artificial seawater (ASW) for the sampler wash.     

Statistical analysis of estuarine profiles: III. Application to nitrate,nitrite and ammonium in the Tamar estuary

Estuarine profiles of nitrate, nitrite and ammonium concentration taken over a period of six years were analysed by a statistical procedure. While nitrate profiles indicate conservative mixing, those of nitrite and ammonium exhibit maxima indicative of an estuarine input. Calculations using an advective analogue suggest that production of nitrite by oxidation throughout the water column would require unduly high populations of nitrifying bacteria (>10⁶ cells l⁻¹). Sedimentary production rates required to sustain the observed nitrite maxima are compatible with combined nitrification and denitrification rates observed elsewhere (1–2μmol m⁻² d⁻¹). The relative displacement of the nitrite and ammonium maxima and the frequent presence of a turbidity maximum in the Tamar estuary suggest that nitrite production in the sediment is probably augmented by water column nitrification in the region of the freshwater/brackish water interface. Simulations on the advective analogue provide circumstantial support for this suggestion.     

国产沸石的改性处理及其在海水硝酸盐氮同位素预处理中的应用

使用多种改性方法对几种国产天然沸石进行改性处理,提高其铵氮吸附率,制备符合海水硝酸盐氮同位素预处理要求的沸石。发现重力筛选可提高沸石铵氮吸附率16%以上,钠改性及酸改性后钠改性可提高沸石铵氮吸附率80%以上,微波改性和超声波改性均可进一步提高沸石铵氮吸附率。改性处理后,几种沸石在酸性条件下对低浓度铵氮吸附率达90%以上,其氮同位素分馏较美国UOP沸石分馏系数更小,且更稳定。改性后的国产沸石更适于海水硝酸盐氮同位素预处理。应用改性后沸石对长江口海域硝酸盐水样进行了分析,结果表明,改性后沸石可以应用于海水中溶解态硝酸盐的氮同位素分析,为海水中溶解态氮的来源问题及循环机理研究等提供了有效信息。     

氮磷比对浙江近岸浮游植物群落结构影响的实验研究

通过室内受控实验方法研究不同氮磷比(N/P)(1:1,4:1,8:1,16:1,32:1,64:1,128:1和256:1)对浙江近岸秋季浮游植物群落结构的影响。结果表明,不同N/P对浮游植物群落的多样性指数、物种组成及演替过程均有显著影响。实验前期(6~12 d),低N/P组(1:1)H',Ma,J和D_r值均偏小,实验结束时(30 d)8:1和16:1组H',J和D_r值小于其他各组。实验初期(6 d),高N/P组(128:1和256:1)对硅藻生长有明显促进作用,实验结束时8:1和16:1组更适合硅藻生长。此外,各组中浮游植物群落都呈硅藻→甲藻的基本演替规律,外界环境条件的改变较之群落初始物种组成对浮游植物群落的演替顺序更具决定性作用。     

Source regions of ammonium nitrate,ammonium sulfate,and natural silicates in the surface aerosols of Moscow oblast

Maps of potential source regions of ammonium nitrate, ammonium sulfate, and natural silicates in the surface aerosols in 2002–2005, 2010, and 2012–2015 are obtained based on measurements of the optical depth of 1–2 μm surface aerosol samples and the analysis results of backward trajectories of air parcels at the Zvenigorod scientific station (ZSS) of the A. M. Obukhov Institute of Atmospheric Physics, Russian Academy of Sciences. The most likely potential source regions of ammonium nitrate are in western and central Europe; those of ammonium sulfate are in southern Ukraine, southern Russia, and northwest Kazakhstan; and those of natural silicates are in the Caspian and Aral regions. The maps of potential source regions are consistent with EMEP fields of surface concentrations of nitrates and sulfates and natural aerosol concentration in PM2.5.     

Identification of Pb sources in Yellow Sea sediments using stable Pb isotope ratios

Rapid economic developments in East Asian countries have inevitably resulted in environmental degradation in the surrounding seas, and concern for both the environment and protection from pollutants is increasing. Identification of sources of contaminants is essential to environmental pollution management. In this study, the provenance of anthropogenic lead (Pb), a major pollutant of Yellow Sea sediments, was determined for river mouth sediments, including those of the Changjiang, Huanghe, Han, and Geum Rivers, and for age-determined shelf core sediments through the measurement of Pb isotope ratios in the HCl-leached fraction using multi-collector inductively coupled plasma-mass spectrometry (MC ICP/MS). Anthropogenic Pb has accumulated in shelf core sediments since 1910, and its isotope ratios were estimated as 0.863–0.866 and 2.119–2.125 for ²⁰⁷Pb/²⁰⁶Pb and ²⁰⁸Pb/²⁰⁶Pb, respectively, from the mixing relationships of the two endmembers. River mouth sediments exhibited enough distinction in anthropogenic Pb isotope ratios to be discriminated: 0.874 (2.144) in the Huanghe, 0.856 (2.129) in the Han, 0.857 (2.122) in the Geum, and 0.854 (2.101) in the Changjiang for ²⁰⁷Pb/²⁰⁶Pb (²⁰⁸Pb/²⁰⁶Pb), respectively. Although isotope ratios of geogenic Pb in sediments dating before 1910 showed narrow ranges (0.842–0.845 and 2.088–2.100 for ²⁰⁷Pb/²⁰⁶Pb and ²⁰⁸Pb/²⁰⁶Pb, respectively), distinct isotope ratios in each core permitted source identification of sediments in the Yellow Sea based on geographic locations and the geogenic Pb of each river. By comparing the isotope ratios of the estimated anthropogenic Pb to source-related materials, the provenances of anthropogenic Pb in Chinese river sediments were presumed to be Chinese coal or ore, which is also a major source of atmospheric particulate Pb. The anthropogenic Pb in the shelf core sediments in the northern Yellow Sea originated from northern Chinese cities such as Beijing and Tianjin through atmospheric pathways. Pb isotope ratios indicated that Pb in Korean river sediments was characteristic of local Korean ores.     

Method for determining stable isotope ratios of dissolved organic carbon in interstitial and other natural marine waters

A procedure is described for the analysis of the stable carbon isotopic composition of dissolved organic carbon (DOC) in natural waters from marine and higher-salinity environments. Rapid (less than 5 min) and complete oxidation of DOC is achieved using a modification of previous photochemical oxidation techniques. The CO₂ evolved from DOC oxidation can be collected in less than 10 min for isotopic analysis. The procedure is at present suitable for oxidation and collection of 1–5 μmol of carbon and has an associated blank of 0.1–0.2 μmol of carbon.Complete photochemical oxidation of DOC standards was demonstrated by quantitative recovery of CO₂ as measured manometrically. Isotopic analyses of standards by photochemical and high-temperature sealed-tube combustion methods agreed to within 0.3.. Photochemical oxidation of DOC in a representative sediment pore-water sample was also quantitative, as shown by the excellent agreement between the photochemical and sealed-tube methods. The δ¹³C values obtained for pore-water DOC using the two methods of oxidation were identical, suggesting that the modified photochemical method is adequate for the isotopically non-fractionated oxidation of pore-water DOC.The procedure was evaluated through an analysis of DOC in pond and pore waters from a hypersaline microbial mat environment. Concentrations of DOC in the water column over the mat displayed a diel pattern, but the isotopic composition of this DOC remained relatively constant (average δ¹³C = −12.4.). Pore-water DOC exhibited a distinct concentration maximum in the mat surface layer, and δ¹³C of pore-water DOC was nearly 8. lighter at 1.5–2.0-cm depth than in the mat surface layer (0–0.5-cm depth). These results demonstrate the effectiveness of the method in elucidating differences in DOC concentration and δ¹³C over biogeochemically relevant spatial and temporal scales. Carbon isotopic analysis of DOC in natural waters, especially pore waters, should be a useful probe of biogeochemical processes in recent environments.     

Simplified procedure for the manual analysis of nitrate in seawater

The use of small volumes of sample and reagents permits the adaption of a copper-coated cadmium column to the manual analysis of nitrate in seawater. The apparatus used for this modification is described.     

Riverine dominance of a nearshore marine demersal food web: evidence from stable isotope and C/N ratio analysis

The Thukela Bank, KwaZulu-Natal, supports a diverse ecosystem and South Africa’s only prawn fishery. Oceanographic studies suggest riverine input is not important for the biology of this system, whereas biological studies suggest the contrary, with prawn catches increasing with increased fluvial run-off. The aim of this study was to determine (i) the importance of riverine and marine organic matter for the Thukela Bank food web; and (ii) whether there are seasonal changes in the Thukela River stable isotope values, and, if so, whether these are reflected in the isotope values of demersal organisms. Estuarine organic matter, sediments and demersal organisms were collected from several sites across the bank in the wet and dry seasons of 2008, 2009 and 2010. Marine particulate organic matter was also collected in 2010 and analysed for δ13C and δ15N, as well as C/N ratios. There were strong seasonal changes in isotopic values of organic matter and fauna, especially faunal δ13C. There was an apparent time-lag in organisms assimilating riverine organic matter isotopic values, with the isotopic signature of demersal organisms reflecting that of riverine organic matter from the previous season, which is likely the result of tissue turnover time. In 2010, Thukela Bank sediment organic matter was of riverine origin and this maintained the demersal food web. We conclude that Thukela River organic matter is an important input to the food web of the Thukela Bank, indicating that any future damming of the catchment area could have serious consequences for this ecosystem.     

Distinctive stable isotope ratios in important zooplankton species in relation to estuarine salinity gradients: Potential tracer of fish migration

To assess the potential of stable isotope ratios as an indicator of fish migration within estuaries, stable isotope ratios in important zooplankton species were analyzed in relation to estuarine salinity gradients. Gut contents from migratory juveniles of the euryhaline marine fish Lateolabrax japonicus were examined along the Chikugo River estuary of the Ariake Sea, which has the most developed estuarine turbidity maximum (ETM) in Japan. Early juveniles in March and April preyed primarily on two copepod species; Sinocalanus sinensis at lower salinities and Acartia omorii at higher salinities. Late juveniles (standard length > 40 mm) at lower salinities preyed exclusively on the mysid Acanthomysis longirostris until July and complementarily on the decapod Acetes japonicus in August. These prey species were collected along the estuary during the spring–summer seasons of 2003 and 2004, and their carbon and nitrogen stable isotope ratios (δ¹³C and δ¹⁵N) were evaluated. The δ¹³C values of prey species were distinct from each other and were primarily depleted within and in close proximity to the ETM (salinity < 10); S. sinensis (−26.6‰) < Acanthomysis longirostris (−23.3‰) < Acartia omorii (−21.1‰) < Acetes japonicus (−18.5‰). The overall gradient of δ¹³C with salinity occurred for all prey species and showed minor temporal fluctuations, while it was not directly influenced by the δ¹³C values in particulate organic matter along the estuary. In contrast to δ¹³C, the δ¹⁵N values of prey species did not exhibit any clear relationship with salinity. The present study demonstrated that δ¹³C has the potential for application as a tracer of fish migration into lower salinity areas including the ETM.     

模拟添加氮对海水溶解无机碳体系的影响

碳和氮作为主要的生源要素对维持海洋生态系的正常运转起着至关重要的作用,碳与氮的变化是相互耦合的且呈双向作用,为探讨海水无机碳与氮的相互作用规律,研究了室内模拟添加硝酸盐对海水无机碳体系pH、溶解无机碳(DIC)、HCO3-、二氧化碳分压(Pco2)的影响。结果表明,在室内培养的条件下,单纯添加硝酸盐(增加至原海水硝酸盐浓度的5-20倍)可引起培养体系浮游生物量的变化,但不能引起海水pH及DIC、HCO-3含量的明显变化,对DIC而言,其变化率仅仅在1%以内,但可导致海水Pco2的相对明显升高,其最终结果导致海水碳汇强度的减弱,碳源强度的增加。     

海洋大气气溶胶的离子色谱分析

对分析方法如采样滤膜的均匀性，滤膜上阴，阳离子的萑萃取率，方法的精密度和检出限等进行了基础性研究，并采用该方法测定了厦门海域大气中的Ｃｌ＾－，Ｆ＾－，ＳＯ＾２－４，ＮＯ＾－３，ＮＨ＾＋４和Ｎａ＾＋。本文法有较高的灵敏度，重现性好，可同时测定多种无机离子，适用于海洋大气气溶胶分析。     

Lipid correction for carbon stable isotope analysis of deep-sea fishes

Stable isotope analysis of fish tissue can aid studies of deep-sea food webs because sampling difficulties severely limit sample sizes of fish for traditional diet studies. The carbon stable isotope ratio (δ¹³C) is widely used in food web studies, but it must be corrected to remove variability associated with varying lipid content in the tissue. A lipid correction has not been determined for any deep-sea fish. These fishes are ideal for studying lipid correction because lipid content varies widely among species. Our objective was to evaluate an application of a mass balance δ¹³C correction to a taxonomically diverse group of deep-sea fishes by determining the effect of lipid extraction on the stable isotope ratios, examining the quality of the model parameters derived for the mass balance correction, and comparing the correction to published results. We measured the lipid extraction effect on the nitrogen stable isotope ratio (δ¹⁵N) and δ¹³C of muscle tissue from 30 North Atlantic species. Lipid extraction significantly increased tissue δ¹⁵N (+0.66‰) and δ¹³C values, but the treatment effect on δ¹³C was dependent on C:N, a proxy for lipid content. We compared the lipid-extracted δ¹³C to the δ¹³C predicted by the mass balance correction using model variables estimated from either all individuals (pooled) or species-by-species or using published values from other species. The correction using the species-by-species approach performed best; however, all three approaches produced corrected values that were generally within 0.5‰ of the measured lipid-free δ¹³C and that had a small over-all bias (<0.5‰). We conclude that a generalized mass balance correction works well for correcting δ¹³C in deep-sea fishes, is similar to that developed for other fishes, and recommend caution when applying a generalized correction to fish with high lipid content (C:N >8).     

Wintertime rates of net primary production and nitrate and ammonium uptake in the southern Benguela upwelling system

The elevated levels of primary productivity associated with eastern boundary currents are driven by nutrient- rich waters upwelled from depth, such that these regions are typically characterised by high rates of nitrate-fuelled phytoplankton growth. Production studies from the southern Benguela upwelling system (SBUS) tend to be biased towards the summer upwelling season, yet winter data are required to compute annual budgets and understand seasonal variability. Net primary production (NPP) and nitrate and ammonium uptake were measured concurrently at six stations in the SBUS in early winter. While euphotic zone NPP was highest at the stations nearest to the coast and declined with distance from the shore, a greater proportion was potentially exportable from open-ocean surface waters, as indicated by the higher specific nitrate uptake rates and f-ratios (ratio of nitrate uptake to total nitrogen consumption) at the stations located off the continental shelf. Near the coast, phytoplankton growth was predominantly supported by ammonium despite the high ambient nitrate concentrations. Along with ammonium concentrations as high as 3.6 µmol l^–1, this strongly suggests that nitrate uptake in the inshore SBUS, and by extension carbon drawdown, is inhibited by ammonium, at least in winter, although this has also been hypothesised for the summer.