Pretreatment Method for Nitrogen Isotopic Analysis of Nitrate in Seawater

A suitable method for the pretreutment of dissolved nitrate samples in seawaters for nitrogen isotopic analysis was established. First, the seawater samples were processed by removing nitrite and amonium. Then Devard's alloy was added in sample for conversion of dissolved nitrate to ammonium. The sample was distilled, and then the ammonium condensate was collected with zeolite. after distillation, the collected condensate was filtered and prepared for determining nitropic values. Some tests of the method were conducted. The distillation condition, the influence of salinity on nitrogen isotopic analysis, absorption of ammonium onto zeolite and an improved method on a large volume of seawater were discussed in this study. The results showed that the distillation step had an average recovery of (104.9 +/- 4.2) % (n = 6) when distillating every 300 mL aliquot of the sample under a strong alkaline condition with 0.5 g devard's alloy and a distillation time of 30 min. The nitrogen isotopic fractionation decreased markedly when salinity was increased from 0% to 0.5%; further increase(1% - 3.5%) showed little effect. The adsorption rate of ammonium onto zeolite had a high yield of (95.96 +/- 1.08) % (n = 6) in average. An improved collection method was used to process a large volume of seawater with several distillations, and had good effect on analysis. The method had been applied to analyze water samples collected from Changjiang estuary. The analytical results indicate that the method is suitable for delta N-15 analysis of dissolved nitrate in seawaters. The present method could provide valuable information about the source and cycle mechanism of dissolved nitrogen in estuary waters.

The nitrogen isotopic composition of dissolved nitrate in the Yangtze River (Changjiang) estuary, China

The nitrogen isotopic composition of dissolved nitrate (delta N-15-NO3-) in surface water of the Yangtze River estuary was determined in four seasons of 2006. delta N-15-NO3- ranged from 0.4 parts per thousand to 6.5 parts per thousand and varied with seasons and geographic regions, reflecting the dynamics of nitrogen cycling in the estuarine ecosystem. delta N-15-NO3- was markedly lower in February than in other seasons and exhibited conservative mixing, which was probably attributed to the NO3- being sourced from the atmospheric deposition and agricultural fertilizer. In the upper estuary, the influence of riverine inputs was important during all surveys. in the turbidity maximum zone, nitrification was found with nitrate depleted in N-15 in May, whereas denitrification resulting in heavy delta N-15-NO3- played an important role in August. More enriched delta N-15-NO3- values coinciding with losses of nitrate concentrations based on the conservative mixing model were found in the adjacent marine area in May, and may reflect obvious phytoplankton assimilation of dissolved nitrate. In this manner, delta N-15-NO3- may be a sensitive indicator of nitrogen sources and biogeochemical processing existing in this estuary in conjunction with the variations of dissolved nitrate and other environmental factors. (C) 2009 Elsevier Ltd. All rights reserved.

Transporting nutrients from sediments: nitrogen and carbon as indicators to measure scavenging ability of a detritivorous fish, Liza haematocheila

The redlip mullet, Liza haematocheila, is a common species in polyculture as a scavenger in China. Feeding on detritus, redlip mullet transports nutrients from sediments up into the water column and converts them into forms that can be utilized by phytoplankton and affects the relative abundance of detritus and dissolved inorganic nutrients to phytoplankton, zooplankton and other fishes. We used nitrogen and carbon as the indicators in this study to measure the scavenging ability, which means intake of nitrogen or carbon by redlip mullet, and the loss of nutrients. Temperature and body weight significantly affected growth nitrogen, faecal nitrogen and faecal carbon. At a certain temperature, the proportion of growth nitrogen or growth carbon increased while the proportion of excretion nitrogen or respiration carbon decreased with increasing body weight.

Geochemical characteristics of nitrogen in the southern Yellow Sea surface sediments

Roles and distributions of various forms of nitrogen in biogeochemical cycling in the southern Yellow Sea surface sediments were investigated. The southern Yellow Sea could be divided into three regions (I, II and III) according to the proportion of fine-grained sediment in > 65%, 35-65% and < 35%, respectively. The ratios of different forms of nitrogen contents between each two of the three regions indicated that the nitrogen contents increased with the proportion of fine-grained sediment increasing. The quanta of exchangeable forms of nitrogen were similar in the three regions, while their releasing time increased from regions I to III, indicating that the cycle of nitrogen in fine-grained sediments was shorter than that in coarse-grained sediments. Nitrogen burial fluxes were also similar in these regions, while the burial efficiency increased from regions I to III. The highest burial efficiency was 30.21% in region III, indicating that more than 70% of nitrogen in the southern Yellow Sea surface sediments could be released to take part in biogeochemical recycling. When all the four forms of exchangeable nitrogen (nitrogen in ion exchangeable form (IEF-N), nitrogen in weak acid extractable form (WAEF-N), nitrogen in strong alkali extractable form (SAEF-N) and nitrogen in strong oxidant extractable form (SOEF-N)) were released to take part in recycling, their potential contributions were 80% (SOEF-N), 11% (IEF-N), 6% (SAEF-N), 3% (WAEF-N) respectively, which showed that SOEF-N was the predominant one, and its contribution to biogeochemical cycling was the highest. (c) 2004 Elsevier B.V. All rights reserved.

Modulatory effects of ammonia-N on the immune system of Penaeus japonicus to virulence of white spot syndrome virus

To study response to white spot syndrome virus (WSSV) under ammonia stress, Penaeus japonicus were exposed to 5 mg l(-1) ammonia-N and challenged orally with WSSV (NW). Controls consisted of an ammonia-N-exposed control group (N), a WSSV-challenged positive control group (W), and an untreated control group (control). Immune parameters measured were total haemocyte count (THC), haemocyte phagocytosis, plasma protein content and haemolymph enzymatic activities for prophenoloxidase (proPO), alkaline phosphatase (ALP), and nitric oxide synthase (NOS). THC and plasma protein had downward trends with time in all treatment groups (NW, N, and W) in contrast to the untreated control group (control). The percentage phagocytosis, NOS activity, and ALP and proPO activity of W and NW decreased initially then increased from 6 to 78 h (except for NOS and ALP, from 6 to 54 h) before declining thereafter until the end of the experiment. Compared with untreated controls (control), there was a downward trend for all measured parameters in the treatment groups (N, NW, and W), but the degree was W > NW > N. WSSV was detected at 78 h postchallenge in both W and NW. In conclusion, 5 mg l(-1) ammonia-N reduced the immunocompetence of P japonicus and may have decreased the virulence of WSSV (C) 2004 Elsevier B.V. All rights reserved.

Characteristics of nitrogen forms in the southern Huanghai Sea surface sediments

The distributions of different forms of nitrogen in the surface sediments of the southern Huanghai Sea are different and affected by various factors. The contents of IEF-N, SOEF-N and TN gradually decrease eastward, and those of SAEF-N northward, while those of WAEF-N westward. Around the seaport of the old Huanghe (Yellow) River, the contents of both SOEF-N and TN are the highest. Among all the factors, the content of fine sediment is the predominant factor to affect the distributions of different forms of nitrogen. The contents of IEF-N, SOEF-N, and TN have visibly positive correlation with the content of fine sediments, and the correlative coefficient is 0.68, 0.58 and 0.71 respectively, showing that the contents of the three forms of nitrogen increase with those of fine sediments. The content of WAEF-N is related to that of fine sediments to a certain extent, with a correlative coefficient of 0.35; while the content of SAEF-N is not related to that of fine sediments, showing that the content of SAEF-N is not controlled by fine grain-size fractions of sediments. In addition, the distributions of different forms of nitrogen are also interacted one another, and the contents of IEF-N and SOEF-N are obviously affected by TN, while those of inorganic nitrogen (WAEF-N, SAEF-N and IEF-N) are not affected by SOEF-N and TN obviously, although they are interacted each other.

Stable carbon and nitrogen isotopic compositions of high molecular weight dissolved organic matter from four US estuaries

High molecular weight dissolved organic matter (HMW-DOM) represents an important component of dissolved organic carbon (DOC) in seawater and fresh-waters. In this paper, we report measurements of stable carbon (delta(13)C) isotopic compositions in total lipid, total hydrolyzable amino acid (THAA), total carbohydrate (TCHO) and acid-insoluble "uncharacterized" organic fractions separated from fourteen HMW-DOM samples collected from four U.S. estuaries. In addition, C/N ratio, delta(13) C and stable nitrogen (delta(15)N) isotopic compositions were also measured for the bulk HMW-DOM samples. Our results indicate that TCHO and THAA are the dominant organic compound classes, contributing 33-46% and 13-20% of the organic carbon in HMW-DOM while total lipid accounts for only <2% of the organic carbon in the samples. In all samples. a significant fraction (35-49%) of HMW-DOM was included in the acid-insoluble fraction. Distinct differences in isotopic compositions exist among bulk samples, the compound classes and the acid-insoluble fractions. Values of delta(13)C and delta(15)N measured for bulk HMW-DOM varied from -22.1 to -30.1parts per thousand and 2.8 to 8.9parts per thousand, respectively and varied among the four estuaries studied as well. Among the Compound classes, TCHO was more enriched in C-13 (delta(13)C = -18.5 to -22.8parts per thousand) compared with THAA (delta(13)C = -20.0 to -29.6parts per thousand) and total lipid (delta(13)C = -25.7 to -30.7parts per thousand). The acid-insoluble organic fractions, in general, had depleted C-13 values (delta(13)C = -23.0 to -34.4parts per thousand). Our results indicate that the observed differences in both delta(13)C and delta(15)N were mainly due to the differences in sources of organic matter and nitrogen inputs to these estuaries in addition to the microbial processes responsible for isotopic fractionation among the compound classes. Both terrestrial sources and local sewage inputs contribute significantly to the HMW-DOM pool in the estuaries studied and thus had a strong influence on its isotopic signatures. Copyright (C) 2004 Elsevier Ltd.

A nitrogen budget of the Changjiang river catchment

Based on 1997-1998 field investigations in the Changjiang river mouth, rain sampling from the river's upper reaches to the mouth, historical data, and relevant literature, the various sources of Total Nitrogen (TN) and Dissolved Inorganic Nitrogen (DIN) in the Changjiang river catchment and N transport in the Changjiang river mouth were estimated. The export fluxes of various form of were mainly controlled by the river runoff, and the export fluxes of NO3-N, DIN and TN in 1998 (an especially heavy flood year) were 1438 103 tonnes (t) yr(-1) or 795.1 kg km(-2) yr(-1) 1746 10(3) t yr(-1) or 965.4 kg km(-2) yr(-1) and 2849 10(3) t yr(-1) or 1575.3 kg km(-2) yr(-1), respectively. The TN and DIN in the Changjiang river came mainly from precipitation, agricultural nonpoint sources, N lost from fertilizer and soil, and point sources of industrial waste and residential sewage discharge, which were about 56.2% and 62.3%, 15.4% and 18.5%, 17.1% and 14.4%, respectively, of the N outflow at the Changjiang river mouth; maximum transport being in the middle reaches.

Seasonal changes in the distribution of dissolved organic nitrogen in coastal and open-ocean waters in the North East Pacific: sources and sinks

The distribution of dissolved organic nitrogen (DON) and nitrate were determined seasonally (winter, spring and summer) during three years along line P, i.e. an E-W transect from the coast of British Columbia, Canada, to Station P (50degreesN, 145degreesW) in the subarctic North East Pacific Ocean. In conjunction, DON measurements were made in the Straits of Juan de Fuca and Georgia within an estuarine system connected to the NE Pacific Ocean. The distribution of DON at the surface showed higher values of 4-17 muM in the Straits relative to values of 4-10 muM encountered along line P, respectively. Along line P, the concentration of DON showed an inshore-offshore gradient at the surface with higher values near the coast. The equation for the conservation of DON showed that horizontal transport of DON (inshore-offshore) was much larger than vertical physical mixing. Horizontal advection of DON-rich waters from the coastal estuarine system to the NE Pacific Ocean was likely the cause of the inshore-offshore gradient in the concentration of DON. Although the concentration of DON was very variable in space and time, it increased from winter to summer, with an average build up of 4.3 muM in the Straits and 0.7 muM in the NE subarctic Pacific. This implied seasonal DON sources of 0.3 mmol N m(-2) d(-1) at Station P and 1.5 mmol N m(-2) d(-1) in the Straits, respectively. These seasonal DON accumulation rates corresponded to about 15-20% of the seasonal nitrate uptake and suggested that there was a small seasonal build up of labile DON at the surface. However, the long residence times of 180-1560 d indicated that the most of the DON pool in surface waters was refractory in two very different productivity regimes of the NE Pacific. (C) 2002 Elsevier Science Ltd. All rights reserved.

Nitrogen isotopic fractionation during a simulated diatom spring bloom: importance of N-starvation in controlling fractionation

N isotope fractionation (epsilon) was first determined during ambient NO3- depletion in a simulated diatom spring bloom. After 48 h of N-starvation, NH4+ was resupplied to the diatoms in small pulses to simulate grazer-produced N and then epsilon was determined. Large variations in epsilon values were observed: from 2.0-3.6 to 14-0 parts per thousand during NO3- and NH4+ uptake, respectively. This is the first study reporting an epsilon value as low as 0 to 2 parts per thousand for NH4+ uptake and we suggest that greater N demand after N-starvation may have drastically reduced NH3 efflux out of the cells. Thus the N status of the phytoplankton and not the ambient NH4+ concentration may be the important factor controlling epsilon, because, when N-starvation increased, epsilon values for NH4+ uptake decreased within 30 h. This study may thus have important implications for interpreting the delta(15)N of particulate N in nutrient-depleted regimes in temperate coastal oceans.

Nitrogen isotope fractionation during nitrate, ammonium and urea uptake by marine diatoms and coccolithophores under various conditions of N availability

Stable isotopes of N provide a new approach to the study of algal production in the ocean, yet knowledge of the isotope fractionation (epsilon) in various oceanic regimes is lacking. Here we report large and rapid changes in isotope composition (delta(15)N) of 2 coastal diatoms and 2 clones (open and coastal) of a coccolithophore grown in the simultaneous presence of nitrate, ammonium and urea under varying conditions of N availability (i.e. N-sufficiency and N-starvation followed by N-resupply) and hence different physiological states, During N-sufficiency, the delta(15)N of particulate organic N (PON) was well reproduced, using a model derived from Rayleigh distillation theory, with constant epsilon similar to that for growth on each individual N source. However, following N-resupply, the variations in delta(15)N(PON) could be well explained only in the case of the open ocean Emiliania huxleyi, with epsilon similar to N-sufficient conditions. It was concluded that the mechanism of isotope fractionation changed rapidly with N availability for the 3 coastal clones. However, in the case of E. huxleyi isolated from the Subarctic Pacific Ocean, no evidence of a change in mechanism was found, suggesting that perhaps open ocean species can quickly recover from N-depleted conditions.

The release behavior of inorganic nitrogen and phosphorus in sediment during disturbance

Sediments and surface water were sampled in a tide flat in the Huiquan Bay, Qingdao, China in January 2004 to simulate the exchange of NH4-N/NO3-N/PO43- between sediments and surface water. A working system was designed with which samples were shaken at 60, 120 and 150 revolutions per minute (r/min). Experiment results show that NH4-N concentration in water at shaking rate of 60 r/min decreased gradually, while at 120 r/min increased gradually. In resuspension, fine-grained sediments contributed most NH4-N to the seawater, followed by medium-grained and coarse-grained sediments. The NO3-N concentration in water had a negative relation, with the shaking rate; the medium-grained sediments contributed more NO3-N to seawater than the coarse- and fine-grained sediments. The PO43- concentration is positively related with the shaking rate, the fine-grained sediments were the main N and P contributor to the seawater, followed by medium- and coarse-grained sediments.

Characteristics of nitrogen forms in the surface sediments of southwestern Nansha Trough, South China Sea

The area of the southwestern Nansha Trough is one of the most productive areas of the southern South China Sea. It is a typical semi-deep sea area of transition from shoal to abyssal zone. To understand distributions and roles of nitrogen forms involved in biogeochemical cycling in this area, contents of nitrogen in four extractable forms: nitrogen in ion exchangeable form (IEF-N), nitrogen in weak acid extractable form (WAEF-N), nitrogen in strong alkali extractable form (SAEF-N) and nitrogen in strong oxidation extractable form (SOEF-N), as well as in total nitrogen content (TN) in surface sediments were determined from samples collected from the cruise in April-May 1999. The study area was divided into three regions (A, B and C) in terms of clay sediment (< 4 mu m) content at < 40%, 40%-60% and > 60%, respectively. Generally, region C was the richest in the nitrogen of all forms and region A the poorest, indicating that the finer the grain size is, the richer the contents of various nitrogen are. The burial efficiency of total nitrogen in surface sediments was 28.79%, indicating that more than 70% of nitrogen had been released and participated in biogeochemical recycling through sediment-water interface.

Changes in nitrogen and phosphorus and their effects on phytoplankton in the Bohai Sea

Systematic studies of the changes in dissolved inorganic nitrogen (DIN) and dissolved inorganic phosphorus (DIP) and their effects on phytoplankton over the last 30 years in the Bohai Sea are presented. The amount of sewage disposal, use of fertilizer and the Huanghe River runoff were found to have a significant influence on the DIN or DIP concentrations in the Bohai Sea over the last 30 years. Moreover, the changes in DIN and DIP resulted in changes in the limiting nutrients of phytoplankton in the Bohai Sea from nitrogen in the early 1980s to nitrogen-phosphorus in the late 1980s, and then to phosphorus after the 1990s. In addition, changes in nitrogen and phosphorus had a significant effect on the phytoplankton community structure. The half saturation constant (K (s)) was used to evaluate the effect of nutrients on the phytoplankton community structure in the Bohai Sea over the last 30 years. Cell abundance percentages of dominant phytoplankton species with high K (s) values for phosphorus and low K (s) values for nitrogen have decreased since the 1980s, while those of dominant phytoplankton species with low K (s) values for phosphorus and high K (s) values for nitrogen increased during this period.

Temporal and spatial variability in nitrogen uptake kinetics during harmful dinoflagellate blooms in the East China Sea

Temporal and spatial variability in the kinetic parameters of uptake of nitrate (NO3-), ammonium (NH4+), urea, and glycine was measured during dinoflagellate blooms in Changjiang River estuary and East China Sea coast, 2005. Karenia mikimotoi was the dominant species in the early stage of the blooms and was succeeded by Prorocentrum donghaiense. The uptake of nitrogen (N) was determined using N-15 tracer techniques. The results of comparison kinetic parameters with ambient nutrients confirmed that different N forms were preferentially taken up during different stages of the bloom. NO3- (V-max 0.044 h(-1); K-s 60.8 mu M-N) was an important N source before it was depleted. NH4+ (V-max 0.049 h(-1); K-s 2.15 mu M-N) was generally the preferred N. Between the 2 organic N sources, urea was more preferred when K. mikimotoi dominated the bloom (V-max 0.020 h(-1); K-s 1.35 mu M-N) and glycine, considered as a dominant amino acid, was more preferred when P. donghaiense dominated the bloom (V-max 0.025 h(-1); K-s 1.76 mu M-N). The change of N uptake preference by the bloom-forming algae was also related to the variation in ambient N concentrations. Published by Elsevier B.V.