δ15N Values Research Articles

Organophosphate diesters (DAPs) and hydroxylated organophosphate flame retardants (HO-OPFRs) as biomarkers of OPFR contamination in a typical freshwater food chain

Organophosphate flame retardants (OPFRs) can rapidly biotransform into two types of metabolites in biota: (1) organophosphate diesters (DAPs) and (2) hydroxylated OPFRs (HO-OPFRs). Therefore, the levels of parent OPFRs alone are not sufficient to indicate OPFR pollution in biological organisms. This study analyzed 12 OPFR metabolites, including 6 DAPs and 6 HO-OPFRs, in a typical freshwater food chain consisted of crucian carp, catfish, mud carp, snakehead, and oriental river prawn. The total concentrations of OPFR metabolites were comparable to those of parent OPFRs, and ranged from 0.65 to 17 ng/g ww. Bis(2-butoxyethyl) 3′-hydroxy-2-butoxyethyl phosphate (14%–77%), di-n-butyl phosphate (DNBP) (6.7%–24%), bis(1-chloro-2-propyl) phosphate (BCIPP) (0.7%–35%), and 1-hydroxy-2-propyl bis(1-chloro-2-propyl) phosphate (BCIPHIPP) (6.0%–24%) were the major OPFR metabolites. Various aquatic species exhibited significant differences in their OPFR metabolite/parent ratios (MPR) (p < 0.05), indicating varying biotransformation potentials of different organisms for various OPFRs. The growth-independent accumulation of tri-n-butyl phosphate (TNBP), tris(chloro-2-propyl) phosphate (TCIPP), triphenyl phosphate, and 2-ethylhexyl diphenyl phosphate in mud carps could be explained by their biotransformation potential. A significant negative correlation was found between the concentration of bis(2-butoxyethyl) phosphate and δ15N values (p < 0.05), with a calculated trophic magnification factor (TMF) of 0.66. Significant positive correlations were observed between BCIPP and TCIPP (R2 = 0.25, p < 0.05), as well as between DNBP and TNBP (R2 = 0.30, p < 0.01), implying that these two DAPs could be used as biomarkers to quantitatively assess TCIPP and TNBP contamination in wild aquatic organisms.

Evidence of isotopic maternal transmission influence on bluefin tuna (Thunnus thynnus) larval growth

Pre-flexion stages of Atlantic bluefin tuna (Thunnus thynnus) larvae were collected in 2014 during the peak of spawning in the two main spawning areas: Gulf of Mexico (GOM) and Mediterranean Sea (MED). We examined daily growth, otolith biometry, and stable isotopes and found that the GOM grew at a faster rate, had larger otoliths, wider daily increments, and significantly lower values of δ15N when compared to the MED. In addition, an intra-population comparative analysis between slow- and fast-growing individuals (deficient vs. optimal growth groups, respectively) showed that optimal growth groups had significantly lower δ15N within each spawning area, implying a direct relationship between growth potential, development, and maternal transmission of isotopic signatures. A third pre-flexion larval group that was aquaculture-reared also exhibited the same pattern to the wild larval groups. In addition, for the first time, we estimated the maternal trophic niches using models developed with field-captured pre-flexion larvae. The estimated maternal trophic niches for the GOM were narrower than the MED, implying differences in the maternal trophodynamics from each nursery area. Overall, the inter-population (GOM vs. MED) and intra-population growth groups (deficient vs. optimal) grew faster and had narrower maternal niches. This study shows the advantages that larval SIA research can aid in the understanding of the trophodynamics of their breeders by examining the trophic relationship of a spawning stock jointly with the development of growth potential in offspring within the same breeding season.

Monitoring hydroecology and climatic variability since ~4.6 ka from palynological, sedimentological and environmental perspectives in an Ox-bow lake, Central Ganga Plain, India

The Ganga-Sai River Interfluve contains several ox-bow lakes in the fertile Central Ganga plains (CGP). A ~2.20-meter deep sedimentary profile obtained near the Chandra Shekhar Azad bird sanctuary (Nawabganj lake-NL) of the CGP was studied to understand the evolution of the ecosystem and climate using pollen/spores, diatoms, testate amoebae, environmental magnetic data,and carbon and nitrogen isotopes. This sedimentary profile is chronologically well-constrained by five radiocarbon (14C) dates. Between 4.6 and 4.4 ka, the sandy sediment and pollen evidence for riparian forest, the absence of aquatic pollen and sponge spicules suggest scant water in the vicinity through the river channel. A semi-closed fluvial ecosystem between 4.4 and 4.2 ka is indicated by testate amoebae, sponge spicules and arboreal pollen. At least two intermittent warm conditions prevailed between 4.6 and 4.2 ka. Between 4.2 and 2.8 ka, high aquatic pollen, diatoms and testate amoebae indicate a lake ecosystem. By ~2.8–0.9 ka the gammoscleres from sponges formed during dry seasons indicate recharging during monsoon as the river shifted. Thereafter, agricultural pollen ( Brassica and Apiaceae) indicates a further shift in the lake boundary exposing land. The highly sandy texture, fluctuating δ13C, δ15N and magnetic mineral values indicate an unstable fluvio-lacustrine deposition inducing hydroecological changes influenced by intermittent about 5–6 humid and dry climatic conditions since ~4.6 ka to present. The calcrete layer in the bottom sediments shows high aridity in CGP between ~5 and 4.6 ka reaching the climax cold-dry event of ~4.2 ka recorded worldwide. The spectral analysis of palynological data from NL and the contemporary Barela Lake, reveals de Vries and Gleissberg cycles of low and high solar irradiance at centennial to multi-centennial scale during the Holocene. The impact on vegetation, sediment depositional dynamics, and shift in river channel was more rapid showing the dominance of ~200 years. periodicity post ~5 ka as compared to ~300 years of dominance prior to this. This centennial timescale is of great speculation for future climate predictions in CGP coupled with the anthropogenic forcings.

Trophic transfer of heavy metals through aquatic food web in the largest mangrove reserve of China

Understanding the mechanism of trophic transfer of heavy metal through the aquatic food web is critical to ecological exposure risk assessments in mangrove ecosystems. Zhanjiang Mangrove National Nature Reserve (ZMNNR) is the largest and biologically richest mangrove reserve in China, but has been exposed to heavy metal pollutants caused by the progressive industrialization and urbanization. We collected a variety of aquatic consumers, and primary producers, as well as sediments from the ZMNNR and analyzed them for heavy metal (Cd, Cr, Cu, Ni, Pb and Zn) concentrations, and for both δ13C and δ15N values to establish the trophic levels.The trophic magnification factors (TMF) of Cd, Cu and Zn are 0.19 (p < 0.01), 0.07 (p < 0.01) and 0.33 (p < 0.05), respectively, indicating significant biodilution in a simplified food web composed of bivalves, crustaceans and fish. There are also potential tendencies of biodilution for Cr, Ni and Pb. Comparison of heavy metals in representative fish and shrimp in the ZMNNR with those in worldwide mangroves indicate a low risk level for aquatic consumers in our ecosystem. Quantitative source tracking is conducted based on principal component analysis and cluster analysis, which indicate that Cr, Ni and Pb are mainly originated from natural geological processes, Cu and Zn from shrimp farming and agriculture activities, and Cd from the deposition of aerosol released by regional metal smelting industry.

Spatially and temporally variable production pathways support the Lake Erie central basin food web

In large freshwater systems, the dominant production pathways supporting food webs are often spatiotemporally variable. We used stable isotope analysis and analysis of covariance (ANCOVA) models to investigate spatial and interannual variation in the dominant production pathways supporting fish consumers within the central basin of Lake Erie. We examined C and N stable isotope ratios of zooplankton, benthic invertebrates, and four species of fish common to nearshore areas of the central basin (yellow perch, Perca flavescens; white perch, Morone americana; rainbow smelt, Osmerus mordax; and round goby, Neogobius melanostomus) using tissue samples collected in 2017 and 2019. δ13C values varied by location consistent with expected baseline differences in nutrient loading (13C was more enriched in the southern region) in two of six ANCOVA models. Furthermore, δ15N values varied with individual fish size and by location in a manner consistent with spatial patterns of nutrient loading from surrounding agricultural landscapes (15N was more enriched in the northern region) and a longitudinal gradient of eutrophication, decreasing from west to east. These patterns were not exhibited by all species and did not necessarily persist across years, suggesting that additional factors (e.g., regional diet differences, river plume dynamics) also contributed to observed δ13C and δ15N variation. We suggest that spatiotemporal variation of stable isotope ratios should be accounted for in studies of trophic basis of production and food web structure in Lake Erie.

Nitrogen uptake by methanotrophic consortia in deep-water gas hydrate-bearing sediments

Methane-consuming microbes inhabiting marine methane seeps have recently been found to have the capacity to assimilate inorganic nitrogen, suggesting a previously unaccounted role in the global nitrogen cycle. Despite ex-situ experimental observations, definitive evidence of this process under in-situ conditions remains elusive, hindering the complete understanding of the controlling factors and magnitude of this process. We present the isotopic variations of organic carbon δ13Corg and total nitrogen δ15N values in two sediment cores collected from the gas hydrate-bearing Håkon Mosby Mud Volcano, SW Barents Sea (72°N, ∼1260 m water depth). We identified a stratigraphic interval containing methane-derived carbonates directly overlying a gas hydrate layer at 67 cm and typified by δ13Corg and δ15N as low as −42.0‰ and 1.2‰, respectively. Stable isotope mixing models confirm in-situ nitrogen uptake by methanotrophic consortia, contributing to up to 49.1 wt% of the local bulk sedimentary organic matter – a finding calling for reevaluation of the role of methane seeps in the oceanic nitrogen cycle.

Equilibrium partitioning and isotopic fractionation of nitrogen between biotite, plagioclase, and K-feldspar during magmatic differentiation

A significant portion of the continental crust is composed of plutonic igneous rocks. However, little is known about the geochemical behaviour of N between the different minerals during magmatic differentiation. To provide new constraints for the behaviour of N during crust formation, we have characterised the geochemistry of nitrogen (N) in the compositionally zoned calc-alkaline pluton at Loch Doon, SW Scotland. We present N concentration and N isotope values for whole-rock data alongside biotite, plagioclase and K-feldspar mineral separates and assess the degree to which these data preserve equilibrium partitioning during magmatic differentiation. We show that whole rock likely inherited its N contents and δ15N signatures from the initial source composition and that this signature is homogenous at a pluton scale. Whilst the whole-rock data are best explained as crust-derived N in the source, the degree of homogenisation across a pluton scale is inconsistent with empirical N diffusivities, ruling out syn-emplacement crustal assimilation as the source of N. Instead, our data suggest a crustal signature inherited from depth associated with the Iapetus subduction zone. At a mineral scale, we find that N preferentially partitions into the feldspars over mica in this system in the order K-feldspar > plagioclase ≈ biotite > quartz, with average mineral–mineral distribution coefficients of DN plagioclase-biotite = 1.3 ± 0.6 and DN Kspar-biotite = 2.8 ± 0.6. Partitioning is accompanied by a large and near constant equilibrium isotope fractionation factor between biotite and both feldspars (averages are Δ15NPlag-Biotite = +7.8 ± 1.2‰ and Δ15NKspar-Biotite = +7.9 ± 1.0‰). In contrast, Δ15NKspar-Plagioclase closely approximates 0‰, where both minerals show δ15N values overlapping with the bulk rock δ15N values. These results show that mica crystallisation generates a 15N-depleted reservoir within plutonic rocks. Moreover, our dataset suggests that feldspars might be a more significant host of N in the igneous portion of Earth’s continental and oceanic crust than previously thought.

Understanding Cassiopea andromeda (Scyphozoa) Invasiveness in Different Habitats: A Multiple Biomarker Comparison

A dual nutrition mode (i.e., mixotrophy) can be advantageous for alien species in a new environment. In Cearà (Brazil), the symbiotic jellyfish Cassiopea andromeda is rapidly spreading under diverse environmental conditions across natural and human-altered coastal habitats, such as mangroves and shrimp farms. Here we report on the trophic ecology of the alien upside-down jellyfish sampled in these two contrasting coastal habitats during the dry (July–October) and rainy (January–April) seasons, investigated by means of organic biomarkers (lipids, carbohydrates) and bulk tissue stable isotope (δ15N and δ13C) analyses. Total lipid content of jellyfish gonads was generally higher in shrimp farms, whereas no significant difference in carbohydrate concentration was found in jellyfish tissues from the two different habitats. Similarly, there were no significant differences in the δ15N values of jellyfish tissues from the two contrasting habitats, whereas the δ13C values were higher in jellyfish from shrimp farms. Overall, the higher carbon-enriched value in aquaculture ponds supports the hypothesis of differences of available food sources compared to the natural mangrove habitats, where food availability exhibits a stronger seasonality. In fact, aquaculture ponds are characterized by human-driven regular food supply, leading to more stable trophic conditions and to enhanced growth, lipid production, and gonadal output of C. andromeda jellyfish. This investigation may contribute to predicting how Cassiopea mixotrophy may contribute to explaining its differential success in different habitats.

Mercury biomagnification and trophic structure patterns in neotropical coastal estuaries impacted by a Chlor-alkali plant in northeast Brazil

The present study reports the concentration of HgT and MeHg in sediments and food webs in estuarine and coastal areas of Northeast Brazil, which have been subject to different impacts, mainly by chlorine and alkali companies. The results showed that the HgT in the sediment varied from 1.7 to 1186 ng g−1 and from 3.3 to 2163 ng g−1 in the organisms. A strong positive relationship between the levels of Hg in the sediment and the organisms was detected. Methylmercury represented 42 to 99% in the muscles of fish and invertebrates. Biomagnification processes were identified for all study areas evaluated according the trophic magnification factor (TMF), which ranged from 3.1 to 12.3. We found significant negative correlations with δ13C values and mercury concentrations and positive correlations between δ15N values and mercury, typical of Hg trophodynamics. This suggests a direct impact of the level of site-specific contamination on local food web and the ecosystem, making continuous monitoring studies necessary to assess food security and the risks that the riverine community is exposed as a result of ingesting mercury contaminated fish.

Linking above and belowground carbon sequestration, soil organic matter properties, and soil health in Brazilian Atlantic Forest restoration

Forest restoration mitigates climate change by removing CO2 and storing C in terrestrial ecosystems. However, incomplete information on C storage in restored tropical forests often fails to capture the ecosystem's holistic C dynamics. This study provides an integrated assessment of C storage in above to belowground subsystems, its consequences for greenhouse gas (GHG) fluxes, and the quantity, quality, and origin of soil organic matter (SOM) in restored Atlantic forests in Brazil. Relations between SOM properties and soil health indicators were also explored. We examined two restorations using tree planting (‘active restoration’): an 8-year-old forest with green manure and native trees planted in two rounds, and a 15-year-old forest with native-planted trees in one round without green manure. Restorations were compared to reformed pasture and primary forest sites. We measured C storage in soil layers (0–10, 10–20, and 20–30 cm), litter, and plants. GHG emissions were assessed using CH4 and CO2 fluxes. SOM quantity was evaluated using C and N, quality using humification index (HLIFS), and origin using δ13C and δ15N. Nine soil health indicators were interrelated with SOM attributes. The primary forest presented the highest C stocks (107.7 Mg C ha−1), followed by 15- and 8-year-old restorations and pasture with 69.8, 55.5, and 41.8 Mg C ha−1, respectively. Soil C stocks from restorations and pasture were 20% lower than primary forest. However, 8- and 15-year-old restorations stored 12.3 and 28.3 Mg ha−1 more aboveground C than pasture. The younger forest had δ13C and δ15N values of 2.1 and 1.7‰, respectively, lower than the 15-year-old forest, indicating more C derived from C3 plants and biological N fixation. Both restorations and pasture had at least 34% higher HLIFS in deeper soil layers (10–30 cm) than primary forest, indicating a lack of labile SOM. Native and 15-year-old forests exhibited higher soil methane influx (141.1 and 61.9 μg m−2 h−1). Forests outperformed pasture in most soil health indicators, with 69% of their variance explained by SOM properties. However, SOM quantity and quality regeneration in both restorations approached the pristine forest state only in the top 10 cm layer, while deeper soil retained agricultural degradation legacies. In conclusion, active restoration of the Atlantic Forest is a superior approach compared to pasture reform for GHG mitigation. Nonetheless, the development of restoration techniques to facilitate labile C input into deeper soil layers (>10 cm) is needed to further improve soil multifunctionality and long-term C storage.

Root trenching and stable isotope analysis uncover trophic links of euedaphic collembola species to mycorrhizal mycelium in pine forests

Ectomycorrhizal fungi are a prominent component of the soil biota of boreal forests, but the role of mycorrhizal mycelium as a food source for soil Collembola remains controversial. We addressed this question in a trenching experiment in young (70 years old) and old (180 years old) stands of Scotch pine, combined with stable isotope analysis. Trenching halved the biomass of ectomycorrhizal mycelium, estimated using in-growth mesh bags. In the young forest, the abundance of two euedaphic Collembola species, Mesaphorura yosiii and Willemia anophthalma, decreased after trenching by 99 and 97% respectively, while in the old forest the abundance of Collembola was not affected. In both forests, trenching reduced δ15N values of the dominant euedaphic species Isotomiella minor and W. anophthalma, indicating a shift in trophic niches. Thus, we obtained convincing evidence of species-specific trophic links of euedaphic Collembola species to the mycelium of mycorrhizal fungi.

Tree-ring N isotopic ratio increased with increasing latitude and decreasing N availability in pine stands across Finland

Since nitrogen (N) is the main forest-growth limiting nutrient in the boreal region, atmospheric N deposition may be an important source of available soil N. The objective of the study was to determine whether the variation in N deposition is reflected in the stem wood N and in its isotopic signatures (variation of 15N/14N ratios relative to atmospheric N2, reported as δ15N values), as well as in current soil properties with a special focus on N cycling. The study material consisted of twelve Scots pine (Pinus sylvestris L.) stands located along the N deposition gradient from south to north in Finland, representing similar site types with relatively unfertile and N-limited soils. Tree-ring N from 5-year segments, spanning a period of 70 years (1951–2020), were examined. Based on the modelled open-place deposition amounts, annual N deposition increased until around 1990 up to 9 kg/ha/year at the southernmost site and 1 kg/ha/year in the north but decreased substantially thereafter. The deposition of N totaled 472 kg/ha at the southernmost and 123 kg/ha at the northernmost site during the 70-year period. δ15N values in tree rings varied between −5.8 and + 1.3 and were higher at the northern than at the southern sites. Tree-ring δ15N ratio correlated negatively with total N, nitrate-N and ammonium-N depositions. The negative correlation still existed when stand age was used as a controlling factor. The correlation also remained negative when the dataset was divided into periods of generally increasing (1951–1990) and decreasing (1991–2020) deposition, or over 30-year age classes, except for the oldest age class (>90-year-old stands). Humus layer pH did not vary much between the sites, but slight signs of decreasing N status existed from south to north. The temporal development of tree-ring δ15N ratio and the amount of N deposition coincided only on some sites after the effect of stand age was controlled. In conclusion, although climate effects cannot be totally excluded, given the same tree species, mycorrhizal type (ectomycorrhiza), site and soil factors, the N isotopic ratios in tree rings distinguished the highest and the lowest N deposition levels. Within small differences in N deposition other factors affecting the ratio may dominate and tracing of the N input from the past is questionable.

Understanding trophic transference role in mercury biomagnification and bioaccumulation in the Atlantic spotted dolphin (Stenella frontalis)

Mercury is a metal of toxicological importance that occurs naturally. However, its concentration can be affected by anthropogenic activities and has the potential to bioaccumulate and biomagnify in food webs. Thus, knowing how its concentration varies along the trophic levels allows us to understand its potential risks to the biota. The present study aimed to investigate mercury transfer through the Stenella frontalis food web in Ilha Grande Bay (IGB), Rio de Janeiro state, Brazil. Samples of muscle and liver of S. frontalis were obtained from carcasses (n = 8) found stranded in the IGB, and its potential prey species were collected in fishing landings in the same Bay (n = 145). Total mercury (THg) concentrations were determined by atomic absorption spectrometry, and the δ15N was determined by an isotope ratio mass spectrometer. To investigate how trophic transfer affects mercury contamination in biota, six linear models were applied between THg logarithmic concentrations and δ15N or trophic position (TP). The trophic magnification factor (TMF) was calculated from each model to estimate the trophic transfer. Mean THg concentration in S. frontalis was higher in the liver than in muscle, but no correlation was found with age and δ15N values. Instead, the hepatic and muscular THg concentrations positively correlated with the trophic position. In the summer, THg concentration, TP, and δ15N values in prey species varied significantly, as well as in the winter, except for THg concentration. All trophic transfer models were significant in both seasons, and the TMF >1. The present study showed that trophic transfer is an essential factor in mercury biomagnification in both seasons but is not the unique driver. Both δ15N and TP could explain mercury trophic transfer, but TP better integrates metabolic diversity and seasonality.

Deconvolving mechanisms of particle flux attenuation using nitrogen isotope analyses of amino acids

AbstractParticulate organic matter settling out of the euphotic zone is a major sink for atmospheric carbon dioxide and serves as a primary food source to mesopelagic food webs. Degradation of this organic matter encompasses a suite of mechanisms that attenuate flux, including heterotrophic metabolic processes of microbes and metazoans. The relative contributions of microbial and metazoan heterotrophy to flux attenuation, however, have been difficult to determine. We present results of compound specific nitrogen isotope analysis of amino acids of sinking particles from sediment traps and size‐fractionated particles from in situ filtration between the surface and 500 m at Ocean Station Papa, collected during NASA EXPORTS (EXport Processes in the Ocean from RemoTe Sensing). With increasing depth, we observe: (1) that, based on the δ15N values of threonine, fecal pellets dominate the sinking particle flux and that attenuation of downward particle flux occurs largely via disaggregation in the upper mesopelagic; (2) an increasing trophic position of particles in the upper water column, reflecting increasing heterotrophic contributions to the nitrogen pool and the loss of particles via remineralization; and (3) increasing δ15N values of source amino acids in submicron and small (1–6 μm) particles, reflecting microbial particle solubilization. We further employ a Bayesian mixing model to estimate the relative proportions of fecal pellets, phytodetritus, and microbially degraded material in particles and compare these results and our interpretations of flux attenuation mechanisms to other, independent methods used during EXPORTS.

Differential feeding habits of the shallow-water hydrothermal vent crab Xenograpsus testudinatus correlate with their resident vent types at a scale of meters

Abstract. The shallow-water hydrothermal vents (SVs) located off Kueishan (KS) Island, Taiwan, are one of the world's most intensively studied vent systems. It has long been known that white vents (WVs) and yellow vents (YVs) differ in the color and composition of the vent plumes. The endemic vent crabs (Xenograpsus testudinatus) are abundant in both vent types, and ovigerous females migrate to the vent periphery with a distance of 100–200 m to release their offspring. However, most research on the vent crabs was associated with WV or unspecified vent areas. To increase our knowledge of the crabs dwelling in other vent types, we compared the feeding habits of the vent crabs living in WV and YV with 2 sampling months. Specifically, we examined the benthic community of WV and YV, the isotopic niche width, and protein expression patterns of the crabs from the two vents at a distance of 100 m and sampled in July and August 2010. The coverage of sessile organisms and low-mobility fauna in WV was more abundant than in YV, based on the survey in August 2010. The δ13C and δ15N values of crabs differed spatially and temporally (multivariate analysis of variance test; p&lt;0.05). The niche width of the vent crabs from YV-August (0.88 ‰2) narrowed substantially compared to the rest, i.e., YV-July (2.94 ‰2), WV-July (2.88 ‰2), and WV-August (3.62 ‰2; p&lt;0.05), respectively. Based on the protein expression patterns, the vent crabs exhibited three groups, i.e., WV-July and YV-July, WV-August, and YV-August, respectively. Our results indicated that the dwelling crabs were associated with their living vent, and within-vent variability was more noticeable in YV compared to WV. We suggested that vent crabs inhabit their resident vent. Even at a scale of meters, trans-vent movement is probably rare as an adaptation to minimize predation risk.

Charring effects on stable carbon and nitrogen isotope values on C4 plants: Inferences for archaeological investigations

Experimental studies demonstrated that charring affects stable isotope values of plant remains. Therefore, it is necessary to consider the impact of charring to reliably interpret δ13C and δ15N values in archaeobotanical remains before using this approach to reconstruct past water management, paleoclimatic changes, and infer paleodietary patterns. Research so far has focused mostly on C3 plants while the charring effect on C4 plants is less understood. This study explored the effects of charring on δ13C, δ15N, %C, %N, and C:N in grains of two C4 species, Sorghum bicolor (L.) Moench (NADP-ME) and Cenchrus americanus (L.) Morrone (heterotypic synonym Pennisetum glaucum (L.) R.Br.) (NAD-ME), grown under the same controlled environmental conditions (watering, light, atmospheric humidity). Sorghum and pearl millet grains were charred from 1 to 3 h at 200–300 °C. Comparing first the uncharred grains, the results show that sorghum has lower δ15N and higher δ13C values than pearl millet. This evidence is also recorded in the charred grains. The charring experiments indicate that the temperature to which the grains are exposed has a higher impact than time on the preservation, mass loss, %C, %N, C:N, and δ13C and δ15N values. Every 50 °C of increase resulted in significant increases of δ15N (+0.37‰) and of δ13C (+0.06‰) values. Increasing the duration of charring to 3 h resulted in significant changes of δ15N (+0.17‰) and no significant changes for δ13C (−0.04‰) values. The average charring effects estimated in our experiment is 0.27‰ (95% CI between −0.02 and 0.56) for δ15N and −0.18‰ (95% CI between −0.30 and −0.06‰) for δ13C. Considering the average values, our data show that pearl millet is more affected by charring than sorghum; however, according to the standard deviations, sorghum shows a greater variability charring effect than pearl millet. This study provides new information to correctly assessing the isotopic values obtained from ancient C4 crops, providing a charring offset specific for C4 plants. Furthermore, it suggests that NAD-ME and NADP-ME species present isotopic differences under the same growing conditions and this must be taken into account in analytical works on ancient C4 crops.

Effects of Thiamethoxam and Fenvalerate Residue Levels on Light-Stable Isotopes of Leafy Vegetables.

Accurate identification of the rational and standardized use of pesticides is important for the sustainable development of agriculture while maintaining a high quality. The insecticides thiamethoxam and fenvalerate and the vegetables spinach, cabbage, and lettuce were used here as study objects. Descriptive analysis and primary reaction kinetic equations were used to analyze the changes in metabolic residues of the two insecticides after different numbers of application in three vegetables. The effects of pesticide residue levels on the δ13C, δ15N, δ2H, and δ18O values of vegetables were analyzed by one-way analysis of variance and correlation analysis. Partial least squares discriminant analysis (PLS-DA) was applied to build discrimination models of the vegetables with different pesticide residues based on stable isotopes. The results showed that the first degradation residues of thiamethoxam and fenvalerate in spinach, cabbage, and lettuce conformed to primary reaction kinetic equations, but the degradation half-lives were long, and accumulation occurred in the second application. The differences in the four stable isotope ratios in the control group of the three vegetables were statistically significant, and two-thirds of the stable isotope ratios in the three vegetables with different numbers of pesticide applications were significantly different. The δ13C and δ15N values of spinach, the δ13C, δ15N, and δ2H values of cabbage, and the δ13C, δ15N, δ2H, and δ18O values of lettuce were significantly correlated with different residues of thiamethoxam and/or fenvalerate applications. The control groups of the three vegetables, spinach-thiamethoxam-first, spinach-thiamethoxam-second, cabbage-thiamethoxam-second, cabbage-fenvalerate-first, and lettuce-thiamethoxam-first, were fully identified by PLS-DA models, while the identification models of other vegetables containing pesticide residues still need to be further improved. The results provide technical support for identifying the rational use of pesticides in vegetables and provide a reference method for guaranteeing the authenticity of green and organic vegetables.

Influence of Seasonal Water Level Fluctuations on Food Web Structure of a Large Floodplain Lake in China

Seasonal shifts in hydrology are known to alter the abundance and diversity of basal production resources and habitats and hence strongly influence the structure and function of river ecosystems. However, equivalent knowledge of natural lake ecosystems in floodplain regions is lacking. Here, we used stable isotope ratios of carbon and nitrogen to assess available primary production sources and consumer taxa during the dry and wet seasons in a large floodplain lake connected to the Yangtze River. Fish species showed distinct δ13C values between two hydrological periods but only small changes in δ15N values. Most of the fish species had higher estimated trophic levels in the dry season, likely indicating greater carnivory. Results of Bayesian mixing models revealed that benthic algae and benthic organic matter (BOM), combined with C3 vegetation, were the principal food sources supporting the biomass of most fish species during the low-water period, whereas benthic algae and seston were the most important carbon sources during the flood period. Overall, these findings demonstrate that seasonal hydrological changes, such as water-level fluctuations, can affect the trophic structure and ecosystem functioning of floodplain lake food webs in the subtropical zone.

What can hippopotamus isotopes tell us about past distributions of C4 grassy biomes on Madagascar?

Societal Impact StatementToday, expansive C4 grassy biomes exist across central, western, and northern Madagascar. Some researchers have argued that the island's now‐extinct pygmy hippopotamuses belonged to a megaherbivore grazing guild that maintained these grasslands prior to human arrival. However, the chemistry of hippo bones indicates that C4 grasses were only a minor part of hippo diet. This, in turn, suggests that C4 grasses were present but not widespread when hippos were alive and that grasses expanded only after Malagasy people shifted from hunting and foraging to agropastoralism approximately 1000 years ago. These results have important implications for environmental reconstructions and biodiversity management.Summary Extinct hippopotamuses (Hippopotamus spp.) were part of Madagascar's megaherbivore guild. Stable carbon (δ13C) and nitrogen (δ15N) isotopes in radiometrically dated bone collagen track spatial and temporal variation in diet and habitat. If hippos helped maintain C4 grassy biomes, then they should have regularly consumed C4 grasses, which have high δ13C values. However, if expansive C4 grassy biomes are anthropogenic, then forests would have been more extensive in the past, and hippos would have predominantly consumed C3 plants with low δ13C values. Nitrogen isotopes can clarify foraging habitat (moist or dry). We assessed δ13C and δ15N values for hippos from different ecoregions of Madagascar and compared these with data for extinct herbivorous lemurs from the same ecoregions. We further explored the effects of wet/dry transitions on isotopic trends for hippos from the central highlands and spiny thicket ecoregions. Carbon isotopes suggest (1) limited C4 consumption by hippos in the central highlands, dry deciduous forest, and succulent woodland ecoregions; and (2) moderate consumption of C4 resources in the spiny thicket. Nitrogen data indicate that hippos foraged in wetter habitats than sympatric lemurs in all regions. Malagasy hippos did not regularly graze C4 grasses in dry, open habitats, even in regions blanketed by C4 grassy biomes today. Malagasy grasses are adapted to grazing and fire, but these are likely ancient adaptations that accompanied grasses when they initially spread to Madagascar. C4 grassy biomes were spatially limited in extent in the past and only expanded after the Late Holocene introduction of domesticated ungulates.

Variation of δ15N in Indian coal, lignite and peat

Nitrogen (N) stable isotope ratio (δ15N) in coal organic matter (OM) provides information on the N source and dominant mechanisms affecting isotopic fractionation during coalification. However, published data on δ15N distribution in coal is rare. The present study is one of the first reports on the δ15N composition of peat, lignite, sub-bituminous and anthracite coals in India and one of the first attempts to understand the processes influencing δ15N composition at different stages of coalification from peat to anthracite. Peats were collected from the western coast of North Andaman Islands and Lake Loktak in Manipur. Plant samples were collected from the peat sampling locations. Cenozoic lignites were collected from Panandhro, Matanomadh, Umarsar and Tadkeshwar mines in Gujarat and Neyveli in Tamil Nadu. Cenozoic sub-bituminous and Permian anthracite coals were collected from Assam and Sikkim, respectively. Variation of δ15N in plants is attributed to the differences in rainfall, plant type and N sources. Lower δ15N values in peats (mean 1.19) compared to the plant samples (mean 2.77) indicate a nonlinear response of δ15N to the relative enrichment or loss of N during peat formation in Lake Loktak and decomposition of OM under anaerobic conditions leading to selective preservation of 14N in the Andaman Islands. The δ15N composition of the studied peat (−1.4–1.6), lignite (−1.4–1.8) and coals (−2.8–5.0) retains their OM source signature. Overall higher δ15N values of Cenozoic lignites compared to the Cenozoic sub-bituminous coal reflects regional differences in climate. Higher δ15N (1.3–5.0) values in Gondwana anthracites reflects the tectonic influence of Himalayan orogeny.