The influence of biopsy site and pregnancy on stable isotope ratios in humpback whale skin.

RATIONALE: Stable isotope analysis (SIA) of free-swimming mysticetes using biopsies is often limited in sample size and uses only one sample per individual, failing to capture both intra-individual variability and the influence of demographic and physiological factors on isotope ratios. METHODS: We applied SIA of δ13C and δ15N to humpback whale (Megaptera novaeangliae) biopsies taken during the foraging season along the western Antarctic Peninsula to quantify intra-individual variation from repeatedly sampled individuals, as well as to determine the effect of biopsy collection site, sex, and pregnancy on isotope ratios. RESULTS: There was substantial variability in δ13C from multiple biopsies taken from the same individuals, though δ15N was much more consistent. Side of the body (left versus right) and biopsy location (dorsal, anterior, ventral, and posterior) did marginally affect the isotopic composition of δ15N but not δ13C. Pregnancy had a significant effect on both δ13C and δ15N, where pregnant females were depleted in both when compared to non-pregnant females and males. CONCLUSIONS: These results indicate that isotopic signatures are influenced by multiple endogenous and exogenous factors and emphasize value in accounting for intra-individual variability and pregnancy status within a sampled population. Placed within an ecological context, the endogenous variability in δ13C observed here may be informative for future isotopic analyses.

Species-specific responses of C and N allocation to N addition: evidence from dual 13C and 15N labeling in three tree species.

Soil nitrogen (N) availability affects plant carbon (C) utilization. However, it is unclear how various tree functional types respond to N addition in terms of C assimilation, allocation, and storage. Here, a microcosm experiment with dual 13C and 15N labeling was conducted to study the effects of N addition (i.e., control, 0 g N kg-1; moderate N addition, 1.68 g N kg-1; and high N addition, 3.36 g N kg-1 soil) on morphological traits, on changes in nonstructural carbohydrates (NSC) in different organs, as well as on C and N uptake and allocation in three European temperate forest tree species (i.e., Acer pseudoplatanus, Picea abies and Abies alba). Our results demonstrated that root N uptake rates of the three tree species increased by N addition. In A. pseudoplatanus, N uptake by roots, N allocation to aboveground organs, and aboveground biomass allocation significantly improved by moderate and high N addition. In A. alba, only the high N addition treatment considerably raised aboveground N and C allocation. In contrast, biomass as well as C and N allocation between above and belowground tissues were not altered by N addition in P. abies. Meanwhile, NSC content as well as C and N coupling (represented by the ratio of relative 13C and 15N allocation rates in organs) were affected by N addition in A. pseudoplantanus and P. abies but not in A. alba. Overall, A. pseudoplatanus displayed the highest sensitivity to N addition and the highest N requirement among the three species, while P. abies had a lower N demand than A. alba. Our findings highlight that the responses of C and N allocation to soil N availability are species-specific and vary with the amount of N addition.

Enhanced dietary reconstruction of Korean prehistoric populations by combining δ13C and δ15N amino acids of bone collagen.

Compound specific stable isotope analysis of amino acids (CSIA-AA) is a powerful tool for determining dietary behaviors in complex environments and improving dietary reconstructions. Here, we conducted CSIA-AA on human (n = 32) and animal (n = 13) remains from two prehistoric archaeological sites (Mumun, Imdang) to assess in more detail the dietary sources consumed by prehistoric Korean populations. Results of estimated trophic position (TP) using Δ15NGlx-Phe show that the Imdang individuals consumed aquatic resources, as well as terrestrial resources. Principal component analysis (PCA) using δ13C and δ15N essential amino acid (EAA) values show that the Imdang humans closely cluster with game birds and terrestrial herbivores, whilst the Mumun humans closely cluster with C4 plants. Quantitative estimation by a Bayesian mixing model (MixSIAR) indicates that the Imdang humans derived a large proportion of their proteins from terrestrial animals and marine fish, whereas the main protein sources for the Mumun humans were C4 plants and terrestrial animals. Additionally, the comparison between the EAA and bulk isotope models shows that there is a tendency to overestimate the consumption of plant proteins when using bulk isotopic data. Our CSIA-AA approach reveals that in prehistoric Korea there were clear differences in human diets through time. This study adds to a growing body of literature that demonstrates the potential of CSIA-AA to provide more accurate estimations of protein consumption in mixed diets than previous bulk isotopic studies.

Diet Affects Egg Laying, Biomass, and Stable Isotope Values in Tetragnathid Spiders.

Riparian tetragnathid spiders are used as biosentinels of aquatic contamination because they are specialized feeders of aquatic emergent insects and are also prey items for terrestrial predators (e.g., birds). Analysis of both trophic position (e.g., stable nitrogen isotopes) and contaminant concentrations are needed to utilize tetragnathids as biosentinels, which can present challenges when collecting enough biomass to reach analytical detection limits, due to their relatively small size. The purpose of this study was to investigate the impacts of a controlled diet source on spider biomass, egg laying and stable isotope values (δ13C and δ15N). Diet significantly influenced the biomass and egg laying of tetragnathids, with some spiders losing up to 50% of their biomass in a single egg-laying event. δ13C had a faster turnover rate in the whole-body of spiders compared to legs, which is important, as spider legs are presently used as surrogates for whole-body δ13C values.

Increase in Agricultural-Derived NHx and Decrease in Coal Combustion-Derived NOx Result in Atmospheric Particulate N-NH4+ Surpassing N-NO3- in the South China Sea.

The atmospheric deposition of anthropogenic active nitrogen significantly influences marine primary productivity and contributes to eutrophication. The form of nitrogen deposition has been evolving annually, alongside changes in human activities. A disparity arises between observation results and simulation conclusions due to the limited field observation and research in the ocean. To address this gap, our study undertook three field cruises in the South China Sea in 2021, the largest marginal sea of China. The objective was to investigate the latest atmospheric particulate inorganic nitrogen deposition pattern and changes in nitrogen sources, employing nitrogen-stable isotopes of nitrate (δ15N-NO3-) and ammonia (δ15N-NH4+) linked to a mixing model. The findings reveal that the N-NH4+ deposition generally surpasses N-NO3- deposition, attributed to a decline in the level of NOx emission from coal combustion and an upswing in the level of NHx emission from agricultural sources. The disparity in deposition between N-NH4+ and N-NO3- intensifies from the coast to the offshore, establishing N-NH4+ as the primary contributor to oceanic nitrogen deposition, particularly in ocean background regions. Fertilizer (33 ± 21%) and livestock (20 ± 6%) emerge as the primary sources of N-NH4+. While coal combustion continues to be a significant contributor to marine atmospheric N-NO3-, its proportion has diminished to 22 (Northern Coast)-35% (background area) due to effective NOx emission controls by the countries surrounding the South China Sea, especially the Chinese Government. As coal combustion's contribution dwindles, the significance of vessel and marine biogenic emissions grows. The daytime higher atmospheric N-NO3- concentration and lower δ15N-NO3- compared with nighttime further underscore the substantial role of marine biogenic emissions.

Mercury bioaccumulation and its relationship with trophic biomarkers in a Mediterranean elasmobranch mesopredator.

Human activity has led to increased concentrations of mercury (Hg) in the world's oceans. Mercury can bioaccumulate and biomagnify in animal tissues via trophic transfer, thus, becoming most pronounced in larger and older predators. Here, we measured Hg concentrations and their relationship with stable isotopes-based proxies of trophic level (δ13C and δ15N values) in multiple tissues of Mustelus spp. from the Mediterranean Sea. We found higher Hg concentrations in muscle than in liver and fin tissues. The relationship between Hg concentrations and δ15N values in muscle suggested repeated foraging for low trophic level and Hg-poor prey, and biomagnification of Hg at higher trophic levels. Seasonal variations in δ13C values could indicate shifts in primary production sources and/or in local prey availability. The HBVSe index suggested no risk to human health, however the safe meal limit recommendations are 4.5 and 2.2 portions per month for adults and children, respectively.

Effects of lipid extraction on stable isotope ratios of carbon and nitrogen in muscles of freshwater fish.

The extraction of lipids by the Folch method from the muscles of all the fish studied led to statistically significant differences in the values of δ15N. At the same time, lipid extraction led to a statistically significant increase in δ13C in pike and roach, and to a statistically insignificant decrease in δ13C in perch and bream. Thus, lipid extraction cannot serve as a universal method of sample preparation for the analysis of the isotopic composition of carbon (13C/12C) and nitrogen (15N/14N) in fish muscles. The differences between the δ13C values in the samples before and after lipid extraction were statistically investigated by different models. It is shown that mathematical correction method models can be used, but the results are depending on the fish types.

Nitrogen isotopic characteristics of aerosol ammonium in a Chinese megacity indicate the reduction from vehicle emissions during the lockdown period.

The role of agricultural versus vehicle emissions in urban atmospheric ammonia (NH3) remains unclear. The lockdown due to the outbreak of COVID-19 provided an opportunity to assess the role of source emissions on urban NH3. Concentrations and δ15N of aerosol ammonium (NH4+) were measured before (autumn in 2017) and during the lockdown (summer, autumn, and winter in 2020), and source contributions were quantified using SIAR. Despite the insignificant decrease in NH4+ concentrations, significantly lower δ15N-NH4+ was found in 2020 (0.6 ± 1.0‰ in PM2.5 and 1.4 ± 2.1‰ in PM10) than in 2017 (15.2 ± 6.7‰ in PM2.5), which indicates the NH3 from vehicle emissions has decreased by∼50% during the lockdown while other source emissions are less affected. Moreover, a reversed seasonal pattern of δ15N-NH4+ during the lockdown in Changsha has been revealed compared to previous urban studies, which can be explained by the dominant effect of non-fossil fuel emissions due to the reductions of vehicle emissions during the lockdown period. Our results highlight the effects of lockdown on aerosol δ15N-NH4+ and the importance of vehicle emissions to urban atmospheric NH3, providing conclusive evidence that reducing vehicle NH3 emissions could be an effective strategy to reduce PM2.5 in Chinese megacities.

Sources and seasonal variations of nitrate in the coastal multiple-aquifer groundwater of Beihai, southern China.

Elevated nitrate (NO3-) loadings in groundwater may cause health effects in drinking water and nutrient enrichment of aquatic ecosystems. To reveal the sources and seasonal variations of NO3- in the coastal groundwater of Beihai, southern China, we carried out hydrochemical and isotopic (δ15N-δ18O in NO3-) investigations in the summer and winter, respectively, concerning multiple-aquifer groundwater, rainwater, seawater, and surface water. The sources of the main elements present in the waters were interpreted by ionic ratios. NO3- sources were identified by combined use of the δ15N values and δ18O values or NO3-/Na+ molar ratios, with estimations of the proportional contribution by the Bayesian stable isotope mixing model. Denitrification was interpreted along the flow paths. The results show groundwater main elements are originated primarily from silicate weathering, and secondarily from anthropogenic inputs and carbonate dissolution. Its qualities are largely affected by seawater intrusion along the coastline. Because of difference in the predominant minerals within the aquifers and in scale and extent of seawater intrusion, the groundwater displays distinct ionic ratio characters. NO3- concentrations are up to 33.9 mg/L, with higher loadings in the plains relative to along the coastline. Soil N, domestic sewage, rainwater, chemical fertilizers, and algae are NO3- sources, with average proportional contributions of 0.255, 0.221, 0.207, 0.202, and 0.116, respectively. In relation to the winter, higher production of NO3- from nitrification of soil N- and algae-derived ammonium induced by higher temperatures in the summer accounts for increases in groundwater NO3- loadings. In the rural areas, elevated loadings of NO3- in the winter may be due to larger infiltration fractions of sewage. Seasonal variations of atmospheric NO3- deposition and farming may also cause the dynamics. Our results improve the understanding of sources and seasonal dynamics of NO3- in coastal groundwater.

Stable isotope analyses of carbon and nitrogen in hair keratin of suspected man-eating wolves from 1880s.

The so-called man-eating wolves of Turku, a pack of three wolves, reportedly killed 22 children in South-Western Finland in 1880-1881. Enormous efforts were carried out to eradicate them. In January 1882 the last remaining wolf was killed. Since then, there has been considerable debate regarding the validity and extent of the man-eating behaviour. This study aims to clarify whether man-eating behaviour can be observed from the remains of these wolves. One of the wolves was mounted in 1882 and is on display at St. Olaf's school in Turku, enabling us to collect hair keratin samples. Additionally, hair keratin was collected from two other suspected man-eaters. We analysed carbon (δ13C) and nitrogen (δ15N) stable isotope values to study the wolf's diet during the last months of its life. Samples from seven temporally concurrent wolves were used to construct reference values. Our analyses indicated that δ15N values of suspected man-eaters were relatively low compared to the reference sample. We could not detect clear trends in isotope ratios associated with potential man-eating behavior. We believe that this lack of distinctive patterns can be explained by the relatively minor role that man-eating played in their overall diet.

Trophic assessment of red seabream (Pagrus major) as a possible bioindicator of human activities in the South Sea of Korea using stable C and N isotopes.

To assess the impact of sand mining on resource utilization by the red seabream (Pagrus major) and the trophic structure of fish assemblages two years after mining activities, we compared stable isotope ratios (δ13C and δ15N) and isotopic niches between aggregated mining and control sites in April and August 2022. Our results showed no spatial differences in the δ13C and δ15N values of red seabream between the sand mining and control sites, suggesting that the mining did not affect their dietary resources. Furthermore, the considerable overlap among fish consumers suggested that the fish food web in mining areas has trophic functions similar to those in natural habitats after mining activities. Overall, our study enhances our understanding of ecosystem conservation and the ecological-based management of coastal areas.

Stable isotope composition of pesticides in commercial formulations: The ISOTOPEST database.

By assessing the changes in stable isotope compositions within individual pesticide molecules, Compound Specific Isotope Analysis (CSIA) holds the potential to identify and differentiate sources and quantify pesticide degradation in the environment. However, the environmental application of pesticide CSIA is limited by the general lack of knowledge regarding the initial isotopic composition of active substances in commercially available formulations used by farmers. To address this limitation, we established a database aimed at cataloguing and disseminating isotopic signatures in commercial formulations to expand the use of pesticide CSIA. Our study involved the collection of 25 analytical standards and 120 commercial pesticide formulations from 23 manufacturers. Subsequently, 59 commercial formulations and 25 standards were extracted, and each of their active substance was analyzed for both δ13C (n = 84) and δ15N CSIA (n = 43). The extraction of pesticides did not cause significant isotope fractionation (Δ13C and Δ15N < 1‰). Incorporating existing literature data, stable carbon and nitrogen isotope signatures varied in a relatively narrow range among pesticide formulations for different pesticides (Δ13C and Δ15N < 10‰) and within different formulations for a single substance (Δ13C and Δ15N < 2‰). Overall, this suggests that pesticide CSIA is more suited for identifying pesticide transformation processes rather than differentiating pesticide sources. Moreover, an inter-laboratory comparison showed similar δ13C (Δ13C ≤ 1.2 ‰) for the targeted substances albeit varying GC-IRMS instruments. Insignificant carbon isotopic fractionation (Δ13C < 0.5‰) was observed after 4 years of storing the same pesticide formulations, confirming their viability for long-term storage at 4 °C and future inter-laboratory comparison exercises. Altogether, the ISOTOPEST database, in open access for public use and additional contributions, marks a significant advancement in establishing an environmentally relevant pesticide CSIA approach.

Uptake patterns for nitrogen and sulfur source by aquatic plants and various nitrogen acquisition strategies: Affected by mining activities.

Understanding the nitrogen and sulfur uptake strategies of mine plants, including sources and preferences for nitrogen forms (ammonium nitrogen (NH4+) vs nitrate nitrogen (NO3-)), is critical to improving understanding of the role of plants in participating in the biogeochemical cycles of nitrogen and sulfur in mining areas. In this study, the stable N and S isotopic compositions of two species of aquatic plants (calamus and reed) in Linhuan mining area were analyzed to determine their absorption strategies for different nitrogen and sulfur sources. The results showed that river water was the largest source of nitrogen and sulfur, contributing 54.6% and 53.9% respectively. NO3- is the main form of nitrogen uptake by reed and calamus, followed by NH4+. In order to adapt to the change of nitrogen form in the environment, reed and calamus tend to absorb and utilize NO3- to maintain their absorption of nitrogen. Mine effluents from mining activities provide at least 12.9% and 16.8% sulfate to reed and calamus respectively, and the effect of mine effluents on reed and calamus sulfur has been underestimated. This study reveals the key factors controlling plant isotope composition, and the use of nitrogen and sulfur isotope composition of aquatic plants can help quantify the level of influence of mining activities, and understand the biogeochemical cycle of nitrogen and sulfur in mining areas.

Enameloid-bound δ15 N reveals large trophic separation among Late Cretaceous sharks in the northern Gulf of Mexico.

The nitrogen isotopic composition (15 N/14 N ratio, or δ15 N) of enameloid-bound organic matter (δ15 NEB ) in shark teeth was recently developed to investigate the biogeochemistry and trophic structures (i.e., food webs) of the ancient ocean. Using δ15 NEB , we present the first nitrogen isotopic evidence for trophic differences between shark taxa from a single fossil locality. We analyze the teeth of four taxa (Meristodonoides, Ptychodus, Scapanorhynchus, and Squalicorax) from the Late Cretaceous (83-84 Ma) Trussells Creek site in Alabama, USA, and compare the N isotopic findings with predictions from tooth morphology, the traditional method for inferring shark paleo-diets. Our δ15 NEB data indicate two distinct trophic groups, with averages separated by 6.1 ± 2.1‰. The lower group consists of Meristodonoides and Ptychodus, and the higher group consists of Scapanorhynchus and Squalicorax (i.e., lamniforms). This δ15 NEB difference indicates a 1.5 ± 0.5 trophic-level separation between the two groups, a finding that is in line with paleontological predictions of a higher trophic level for these lamniforms over Meristodonoides and Ptychodus. However, the δ15 NEB of Meristodonoides is lower than suggested by tooth morphology, although consistent with mechanical tests suggesting that higher trophic-level bony fishes were not a major component of their diet. Further, δ15 NEB indicates that the two sampled lamniform taxa fed at similar trophic levels despite their different inferred tooth functions. These two findings suggest that tooth morphology alone may not always be a sufficient indicator of dietary niche. The large trophic separation revealed by the δ15 NEB offset leaves open the possibility that higher trophic-level lamniforms, such as those measured here, preyed upon smaller, lower trophic-level sharks like Meristodonoides.

Changes in urinary stable nitrogen isotope ratios during controlled short-term energy deficit: a proof-of-principle analysis.

PURPOSE: Stable isotope ratios of nitrogen (δ15N) have previously been shown to increase in human hair during periods of catabolism. The goal of this study was to assess changes in δ15N in urinary urea (δ15Nurea) and Δ15N during a short-term controlled energy deficit. METHODS: We analyzed samples from 6 recreationally active men (25 ± 1 years, BMI: 23.5 ± 0.6 kg/m2) who participated in a repeated measures cross-over study involving 4 days of energy deficit (ED, ~ 15 kcal/kg FFM) without and with exercise (ED-EX, ED + EX) and control conditions in energy balance (CON-EX, CON + EX). δ15Nurea was analyzed from urine samples, and Δ15N was calculated as δ15Nurea-δ15Ndiet, with δ15Ndiet obtained from diet prescriptions. RESULTS: δ15Nurea was significantly elevated in ED-EX (4.4 ± 0.2‰) when compared to CON-EX (3.7 ± 0.1‰; p = 0.026) and CON + EX (3.34 ± 0.13‰, p = 0.001). As a consequence, Δ15N was positive in ED-EX (0.2 ± 0.2‰) and remained negative in ED + EX (- 0.6 ± 0.5‰), CON-EX (- 1.0 ± 0.2) and CON + EX (- 1.1 ± 0.2). Differences in Δ15N were significant between ED-EX and CON-EX (p = 0.005) and ED-EX and CON + EX (p = 0.006). CONCLUSION: Our results suggest that δ15Nurea and subsequently Δ15N are responsive to a short-term energy deficit, likely due to increased amino acid oxidation to meet energy demands and preferable elimination of 14N.

Potential impacts of coal mining activities on nitrate sources and transport in a karst river basin in southwest China.

Typical sources of nitrate pollution in the fragile ecological environment of karst areas, such as agricultural production activities and domestic sewage, have long attracted serious concern. However, coal development can play an equally significant role in releasing the nitrogen fixed in coal into surface watersheds in the form of nitrate, nitrite, or ammonia, consequently threatening the water quality of surface water systems in mining areas. In this study, a typical karst surface watershed system affected by coal mining activities was selected for an in-depth investigation with the aim of realistically assessing the potential contribution of coal mining to nitrogen pollution. The results reveal increasingly concerning nitrate pollution from August 2020 to November 2021 in the Huatan River watershed under the influence of anthropogenic activities, especially mining development and agricultural production. Given that the nitrogen and oxygen isotope compositions of nitrate do not support the presence of denitrification, the variation in the NO3-/Cl- ratio and the relatively stable Cl- concentration may be a reflection of nitrification. Although the leaching of atmospheric precipitation on the strata in the basin promoted the release of nitrogen associated with coal mining, the higher rate of nitrogen cycling in the oligotrophic mine water environment limited the contribution of coal mining to nitrogen pollution in the surface watershed. Specifically, the contribution of coal mining activities to nitrogen pollution in surface karst river is mainly NH4+-N, which contributes 10% or less to the nitrate input to the waters of the Huatan River. The findings thus highlight the necessity of further uncovering the geochemical cycling process of nitrogen during the transport of mine water in the coal mining environment.

Implication of self-organizing map, stable isotopes combined with MixSIAR model for accurate nitrogen control in a well-protected reservoir.

Nitrogen pollution and eutrophication in reservoirs is a global environmental geochemical concern. Occasional algal blooms still exist in reservoirs that have undergone pollution treatment. The lack of quantitative evidence of nitrogen sources and fate limits long-term stable ecological safety management. This work applied an approach integrated zonal mapping, stable isotopes (δ18OH2O, δ15Nnitrate, δ18Onitrate, and δ13C-DIC) and a Bayesian isotope model to analyze regional and seasonal differences in the contribution and sources of nitrogen to a well-protected reservoir. The values of δ18Onitrate and the positive relationship between NO3- and δ13C-DIC suggested that nitrification was the primary NO3- production in the rivers. While Denitrification was present at only a few sites. Results of the MixSIAR model coupled the NO3-/Cl- indicator revealed that the domestic sewage contributed high riverine NO3- loading (68.6 ± 10.6 %) in the dry season. In the wet season, the main nitrate sources of upper watershed were ammonia and carbamide fertilizers (47.5 % and 40.3 %). While the domestic sewage was still the major contributor of downstream region (a dense residential area), indicating possible problems with rainwater and sewage drainage networks. The results implied that the colleting and treatment of sewages were the priority in downstream region, and non-point source pollution control and wastewater treatment plant upgrading were essential to control nitrate pollution in the two upstream regions. These findings provide new insights into precise nitrogen pollution traceability and identification of treatment priorities in the sub-region, and promote the management other well-protected watershed in similar need of further nitrogen contamination control.

Influence of the intensive mariculture on coastal sedimentary organic matter: Insight from size-fractionated particles.

A portion of carbon produced from shellfish and kelp cultivation is buried in sedimentary environment, and mariculture carbon sequestration potential is an important part of marine carbon sink and has attracted worldwide attention. Total organic carbon (TOC), total nitrogen (TN) and their stable isotopes (δ13C and δ15N), as well as the mass distribution of these size-fractionated particles were determined in order to study the distribution and sources of TOC in Sanggou Bay. Results showed that sediment organic matter has complex sources from kelp (30.4 %), marine phytoplankton (25.6 %), shellfish (23.7 %), terrestrial input (20.3 %), and mariculture activities of shellfish and kelp was the major component in surface sediment. Approximately 44-69 % of TOC was associated with the 16-32 µm fraction. Low δ13C (-22.1 to -15.1‰) and high δ15N (5.0-5.7‰) were observed in fine particles (<16 µm), indicating relatively high contribution of marine phytoplankton and mariculture derived organic carbon. On the contrary, relatively higher δ13C (-20.2 to -9.2‰) and lower values δ15N (-4.7 to 5.2‰) in coarse particles (>32 µm) suggested that sedimentary organic carbon might be influenced by some additional sources from terrestrial input or seaweed. The mass distribution, δ13C and δ15N of size-fractionated particles in sediments indicated that sediment was obviously redistributed under the condition of mariculture, and further suggested that mariculture derived organic matter have modified the distribution and sources of sedimentary organic matter. This study provided great insight into distribution and source of sedimentary organic carbon from the perspective of size-fractionated particles in mariculture area.

Nitrate budget of a terrestrial-to-marine continuum in South China: Insights from isotopes and a Markov chain Monte Carlo model.

Anthropogenic nitrate (NO3-) production has been increasing and is exported to the ocean via river networks, causing eutrophication and ecological damage. While studies have focused on river NO3- pollution, what has been lacking is the quantification of the sources of NO3- in coastal rivers. This study applied the dual isotopes (δ15N/δ18O-NO3-) to quantify the sources and their fluxes of NO3- in two inflow rivers of the Qinzhou Bay. By adding our results to the NO3- source apportionment in Qinzhou Bay, we, for the first time, established the NO3- budgets of the terrestrial-to-marine continuum in both high- and low-flow seasons. We quantitatively showed the direct and indirect roles (e.g., the stimulation of nitrification by sewage ammonium-NH4+) of terrestrial sources in driving the high NO3- loading in the estuary. The results highlighted the necessity to consider coastal rivers and estuary as a whole, which could shed light on the effective reduction of NO3- pollution in coastal environments.

Effects of sampling time and location on the geographical origin traceability of protected geographical indication (PGI) Hongyuan yak milk: Based on stable isotope ratios.

Hongyuan yak milk is a protected geographical indication (PGI) product of rich nutritional value, which is popular among consumers. Stable isotope ratio analysis (SIRA) is an effective way to protect the authenticity of the geographical origin of PGI products, and it is crucial to study the factors affecting stable isotopes. Firstly, we proved that the SIRA could be used to identify the geographical origin of Hongyuan yak milk, and that the identification accuracy in combination with δ13C and δ18O was 100 %. Secondly, we analyzed the effect of sampling selection on the stable isotopes of Hongyuan yak milk in practical applications, which showed that sampling time influenced the δ13C, δ2H, and δ18O, while the sampling locations did not. There were interactions between the effect of sampling time and location on δ2H and δ18O. These results provide a reliable method for identifying PGI products and also provide new guidance on sampling models.