Nitrogen isotope effects in the assimilation of inorganic nitrogenous compounds by marine diatoms

Nitrogen isotope fractionation in the assimilation of inorganic nitrogenous compounds was studied using marine diatoms (Phaeodactylum tricornutum and Chaetoceros sp.). The isotopic composition (δ¹⁵N) of the diatoms ranged from 7 to ‐18‰ relative to that of the nitrogen source, i.e., ammonium, nitrite, or nitrate. When the growth was light‐limited, the isotope fractionation in nitrate assimilation was inversely correlated with the growth rate. The highest fractionation factor of 1.016 was obtained when the growth rate was as low as 0.025 day^‐1. Fractionation was negligible when the growth, rate was higher than 1 day^‐1. A steady‐state kinetic model was applied to explain the isotope fractionation in nitrate assimilation. The nitrogen isotope fractionation primarily takes place at the step of N‐O bond breaking in nitrate reduction to nitrite. The extent of the isotope fractionation associated with the nitrate uptake is very small, and barely exceeds the limit of detection.     

Isotopologue profiling of the listerial N‐metabolism

The nitrogen (N‐) sources and the relative contribution of a nitrogenous nutrient to the N‐pool of the gram‐positive pathogen Listeria monocytogenes are largely unknown. Therefore, ¹⁵N‐isotopologue profiling was established to study the N‐metabolism of L. monocytogenes. The pathogen was grown in a defined minimal medium supplemented with potential ¹⁵N‐labeled nutrients. The bacteria were harvested and hydrolysed under acidic conditions, and the resulting amino acids were analysed by GC‐MS, revealing ¹⁵N‐enrichments and isotopomeric compositions of amino acids. The differential ¹⁵N‐profiles showed the substantial and simultaneous usage of ammonium, glutamine, methionine, and, to a lower extent, the branched‐chain amino acids valine, leucine, and isoleucine for anabolic purposes, with a significant preference for ammonium. In contrast, arginine, histidine and cysteine were directly incorporated into proteins. L. monocytogenes is able to replace glutamine with ethanolamine or glucosamine as amino donors for feeding the core N‐metabolism. Perturbations of N‐fluxes caused by gene deletions demonstrate the involvement of ethanolamine ammonia lyase, and suggest a role of the regulator GlnK of L. monocytogenes distinct from that of Escherichia coli. The metabolism of nitrogenous nutrients reflects the high flexibility of this pathogenic bacterium in exploiting N‐sources that could also be relevant for its proliferation during infection.     

NMR analysis of plant nitrogen metabolism

The analysis of primary and secondary nitrogen metabolism in plants by nuclear magnetic resonance (NMR) spectroscopy is comprehensively reviewed. NMR is a versatile analytical tool, and the combined use of ¹H, ²H, ¹³C, ¹⁴N and ¹⁵N NMR allows detailed investigation of the acquisition, assimilation and metabolism of nitrogen. The analysis of tissue extracts can be complemented by the in vivo NMR analysis of functioning tissues and cell suspensions, and by the application of solid state NMR techniques. Moreover stable isotope labelling with ²H-, ¹³C- and ¹⁵N-labelled precursors provides direct insight into specific pathways, with the option of both time-course and steady state analysis increasing the potential value of the approach. The scope of the NMR method, and its contribution to studies of plant nitrogen metabolism, are illustrated with a wide range of examples. These include studies of the GS/GOGAT pathway of ammonium assimilation, investigations of the metabolism of glutamate, glycine and other amino acids, and applications to tropane alkaloid metabolism. The continuing development of the NMR technique, together with potential applications in the emerging fields of metabolomics and metabolic flux analysis, leads to the conclusion that NMR will play an increasingly valuable role in the analysis of plant nitrogen metabolism.     

Nitrogen stable isotope ratios in surface sediments, epilithon and macrophytes from upland lakes with differing nutrient status

1. Thirty small upland lakes in Cumbria, Wales, Scotland and Northern Ireland were each visited once during June and July 2000. From each lake, samples of surface sediment epilithon, macrophytes and total dissolved nitrogen (TDN) were collected for nitrogen stable isotope analysis. As part of a wider programme, samples were also collected for chemical analysis and bioassays. 2. Considerable variation was found in δ¹⁵N values in all measured nitrogen compartments. Some regional variation was evident but was generally weak. Sediment and epilithon δ¹⁵N were positively correlated with δ¹⁵N of TDN, suggesting that baseline nitrogen isotope ratios influence those in some organic matter compartments in the lakes. 3. Sediment δ¹⁵N was higher when inorganic nitrogen concentration in the water was low, possibly reflecting reduced isotope fractionation under these conditions. However, this was not the case for epilithon or macrophytes. Sediment δ¹⁵N values were also negatively related to annual nitrogen deposition. 4. Sediment, epilithon and macrophyte δ¹⁵N values all showed significant relations to nutrient limitation in the lakes as determined by algal bioassays. We suggest that sediment δ¹⁵N might be developed as a simple integrating measure of the degree of nitrogen limitation in lakes.     

A distinctive latitudinal trend of nitrogen isotope signature across urban forests in eastern China

Rapid urbanization has greatly altered nitrogen (N) cycling from regional to global scales. Compared to natural forests, urban forests receive much more external N inputs with distinctive abundances of stable N isotope (δ¹⁵N). However, the large-scale pattern of soil δ¹⁵N and its imprint on plant δ¹⁵N remain less well understood in urban forests. By collecting topsoil (0–20 cm) and leaf samples from urban forest patches in nine large cities across a north–south transect in eastern China, we analyzed the latitudinal trends of topsoil C:N ratio and δ¹⁵N as well as the correlations between tree leaf δ¹⁵N and topsoil δ¹⁵N. We further explored the spatial variation of topsoil δ¹⁵N explained by corresponding climatic, edaphic, vegetation-associated, and anthropogenic drivers. Our results showed a significant increase of topsoil C:N ratio towards higher latitudes, suggesting lower N availability at higher latitudes. Topsoil δ¹⁵N also increased significantly at higher latitudes, being opposite to the latitudinal trend of soil N availability. The latitudinal trend of topsoil δ¹⁵N was mainly explained by mean annual temperature, mean annual precipitation, and atmospheric deposition of both ammonium and nitrate. Consequently, tree leaf δ¹⁵N showed significant positive correlations with topsoil δ¹⁵N across all sampled plant species and functional types. Our findings reveal a distinctive latitudinal trend of δ¹⁵N in urban forests and highlight an important role of anthropogenic N sources in shaping the large-scale pattern of urban forest ¹⁵N signature.     

δ15N of zooplankton species in subarctic lakes in northern Sweden: effects of diet and trophic fractionation

1. To assess the use of stable nitrogen isotopes (δ¹⁵N) for reconstructing trophic relationships in planktonic food webs, crustacean zooplankton species and pelagic dissolved and particulate matter were analysed in 14 subarctic lakes in northern Sweden. The lakes are situated along an altitudinal gradient and show a substantial variation in nutrient content and energy mobilization by bacterioplankton and phytoplankton. 2. The δ¹⁵N of dissolved and particulate matter was comparatively low, suggesting efficient N recycling and low losses of depleted N from the pelagic zone of these unproductive lakes. 3. Copepods had a systematically higher δ¹⁵N than cladocerans, with an average difference of 3.1–4.9‰ within lakes, implying different trophic positions of the two groups. Comparisons of nitrogen pools and energy fluxes suggest that the low cladoceran δ¹⁵N was a result of feeding on bacteria. 4. The difference in δ¹⁵N between copepods and cladocerans declined with decreasing bacterioplankton production among lakes, due either to increasing trophic isotope fractionation or decreasing relative importance of bacteria in the diet of cladocerans.     

Isotope composition of carbon and nitrogen in tissues and organs of <Emphasis Type="Italic">Betula pendula</Emphasis>

Ratios of ¹³С/¹²C and ¹⁵N/¹⁴N isotopes were identified in different parts and organs of drooping birch (Betula pendula Roth) in preforest-steppe and pine-birch forests of the Middle Urals by mass spectrometry. The data were analyzed and interpreted from the perspective of biochemical processes of carbon and nitrogen metabolism in the leaf, cambial tissue, trunk wood, branches, roots, and in the soil. The lighter isotopic composition of carbon is characteristic for the leaves, trunk cambium as well as fine (<2 mm) roots. The trunk wood is characterized by the basal trend for ¹³C enrichment. The heavier carbon isotopic composition inversely related to metabolic activity of organs and tissues, in addition, ¹³С/¹²C ratio corresponds to the nitrogen content in the organs and tissues, indicating the metabolic control of carbon fractionation in woody plants. The isotopic composition of nitrogen in the aboveground parts of the plant (leaves, trunk cambium, wood) and in the medium and fine roots was significantly depleted in ¹⁵N (δ¹⁵N varies from 0 to–3‰), while main roots (δ¹⁵N = 0.6 ‰) and soil (δ¹⁵N = 2.4–6.7‰) were more enriched. The ratio of stable isotopes of carbon and nitrogen is an integrating index of carbon and nitrogen metabolism in plants.     

Variation in gut microbial contribution of essential amino acids to host protein metabolism in a wild small mammal community

Herbivory is a dominant feeding strategy among animals, yet herbivores are often protein limited. The gut microbiome is hypothesized to help maintain host protein balance by provisioning essential macromolecules, but this has never been tested in wild consumers. Using amino acid carbon (δ¹³C) and nitrogen (δ¹⁵N) isotope analysis, we estimated the proportional contributions of essential amino acids (AA_ESS) synthesized by gut microbes to five co-occurring desert rodents representing herbivorous, omnivorous and insectivorous functional groups. We found that herbivorous rodents occupying lower trophic positions (Dipodomys spp.) routed a substantial proportion (~40%–50%) of their AA_ESS from gut microbes, while higher trophic level omnivores (Peromyscus spp.) and insectivores (Onychomys arenicola) obtained most of their AA_ESS (~58%) from plant-based energy channels but still received ~20% of their AA_ESS from gut microbes. These findings empirically demonstrate that gut microbes play a key functional role in host protein metabolism in wild animals.     

Natural Isotopic Signatures of Variations in Body Nitrogen Fluxes: A Compartmental Model Analysis

Body tissues are generally ¹⁵N-enriched over the diet, with a discrimination factor (Δ¹⁵N) that varies among tissues and individuals as a function of their nutritional and physiopathological condition. However, both ¹⁵N bioaccumulation and intra- and inter-individual Δ¹⁵N variations are still poorly understood, so that theoretical models are required to understand their underlying mechanisms. Using experimental Δ¹⁵N measurements in rats, we developed a multi-compartmental model that provides the first detailed representation of the complex functioning of the body''s Δ¹⁵N system, by explicitly linking the sizes and Δ¹⁵N values of 21 nitrogen pools to the rates and isotope effects of 49 nitrogen metabolic fluxes. We have shown that (i) besides urea production, several metabolic pathways (e.g., protein synthesis, amino acid intracellular metabolism, urea recycling and intestinal absorption or secretion) are most probably associated with isotope fractionation and together contribute to ¹⁵N accumulation in tissues, (ii) the Δ¹⁵N of a tissue at steady-state is not affected by variations of its P turnover rate, but can vary according to the relative orientation of tissue free amino acids towards oxidation vs. protein synthesis, (iii) at the whole-body level, Δ¹⁵N variations result from variations in the body partitioning of nitrogen fluxes (e.g., urea production, urea recycling and amino acid exchanges), with or without changes in nitrogen balance, (iv) any deviation from the optimal amino acid intake, in terms of both quality and quantity, causes a global rise in tissue Δ¹⁵N, and (v) Δ¹⁵N variations differ between tissues depending on the metabolic changes involved, which can therefore be identified using simultaneous multi-tissue Δ¹⁵N measurements. This work provides proof of concept that Δ¹⁵N measurements constitute a new promising tool to investigate how metabolic fluxes are nutritionally or physiopathologically reorganized or altered. The existence of such natural and interpretable isotopic biomarkers promises interesting applications in nutrition and health.     

Responses to K deficiency and waterlogging interact via respiratory and nitrogen metabolism

K deficiency and waterlogging are common stresses that can occur simultaneously and impact on crop development and yield. They are both known to affect catabolism, with rather opposite effects: inhibition of glycolysis and higher glycolytic fermentative flux, respectively. But surprisingly, the effect of their combination on plant metabolism has never been examined precisely. Here, we applied a combined treatment (K availability and waterlogging) to sunflower (Helianthus annuus L.) plants under controlled greenhouse conditions and performed elemental quantitation, metabolomics, and isotope analyses at different sampling times. Whereas separate K deficiency and waterlogging caused well‐known effects such as polyamines production and sugar accumulation, respectively, waterlogging altered K‐induced respiration enhancement (via the C₅‐branched acid pathway) and polyamine production, and K deficiency tended to suppress waterlogging‐induced accumulation of Krebs cycle intermediates in leaves. Furthermore, the natural ¹⁵N/¹⁴N isotope composition (δ¹⁵N) in leaf compounds shows that there was a change in nitrate circulation, with less nitrate influx to leaves under low K availablity combined with waterlogging and more isotopic dilution of lamina nitrates under high K. Our results show that K deficiency and waterlogging effects are not simply additive, reshape respiration as well as nitrogen metabolism and partitioning, and are associated with metabolomic and isotopic biomarkers of potential interest for crop monitoring.     

Effects of decomposition and storage conditions on the δ13C and δ15N isotope values of killer whale (Orcinus orca) skin and blubber tissues

Several different factors in the collection and preservation of whale skin and blubber samples were examined to determine their effect on the results obtained by stable nitrogen and carbon isotope (δ¹⁵N and δ¹³C) analysis. Samples of wet killer whale skin retained their original stable isotope values for up to 14 d at 4°C or lower. However, decomposition significantly changed the δ¹⁵N value within 3 d at 20°C. Storage at ?20°C was as effective as ?80°C for the preservation of skin and blubber samples for stable isotope analysis for at least a year. By contrast, once a skin sample had been freeze‐dried and lipid extracted, the stable isotope values did not change significantly when it was stored dry at room temperature for at least 12 mo. Preservation of whale skin samples for a month in DMSO‐salt solution, frozen or at room temperature, did not significantly change the δ¹⁵N and δ¹³C values of lipid extracted tissues, although the slight changes seen could influence results of a study if only small changes are expected.     

Comparative response of δ13C, δ18O and δ15N in durum wheat exposed to salinity at the vegetative and reproductive stages

This study compared the performance of the stable isotope composition of carbon (δ¹³C), oxygen (δ¹⁸O) and nitrogen (δ¹⁵N) by tracking plant response and genotypic variability of durum wheat to different salinity conditions. To that end, δ¹³C, δ¹⁸O and δ¹⁵N were analysed in dry matter (dm) and the water‐soluble fraction (wsf) of leaves from plants exposed to salinity, either soon after plant emergence or at anthesis. The δ¹³C and δ¹⁸O of the wsf recorded the recent growing conditions, including changes in evaporative conditions. Regardless of the plant part (dm or wsf), δ¹³C and δ¹⁸O increased and δ¹⁵N decreased in response to stress. When the stress conditions were established just after emergence, δ¹⁵N and δ¹³C correlated positively with genotypic differences in biomass, whereas δ¹⁸O correlated negatively in the most severe treatment. When the stress conditions were imposed at anthesis, relationships between the three isotope signatures and biomass were only significant and positive within the most severe treatments. The results show that nitrogen metabolism, together with stomatal limitation, is involved in the genotypic response to salinity, with the relative importance of each factor depending on the severity and duration of the stress as well as the phenological stage that the stress occurs.     

Foliar and soil δ15N values reveal increased nitrogen partitioning among species in diverse grassland communities

Plant and soil nitrogen isotope ratios (δ¹⁵N) were studied in experimental grassland plots of varying species richness. We hypothesized that partitioning of different sources of soil nitrogen among four plant functional groups (legumes, grasses, small herbs, tall herbs) should increase with diversity. Four years after sowing, all soils were depleted in ¹⁵N in the top 5 cm whereas in non‐legume plots soils were enriched in ¹⁵N at 5–25 cm depth. Decreasing foliar δ¹⁵N and Δδ¹⁵N (= foliar δ¹⁵N ? soil δ¹⁵N) values in legumes indicated increasing symbiotic N₂ fixation with increasing diversity. In grasses, foliar Δδ¹⁵N also decreased with increasing diversity suggesting enhanced uptake of N depleted in ¹⁵N. Foliar Δδ¹⁵N values of small and tall herbs were unaffected by diversity. Foliar Δδ¹⁵N values of grasses were also reduced in plots containing legumes, indicating direct use of legume‐derived N depleted in ¹⁵N. Increased foliar N concentrations of tall and small herbs in plots containing legumes without reduced foliar δ¹⁵N indicated that these species obtained additional mineral soil N that was not consumed by legumes. These functional group and species specific shifts in the uptake of different N sources with increasing diversity indicate complementary resource use in diverse communities.     

Nitrogenous compounds stimulate glucose‐derived acid production by oral Streptococcus and Actinomyces

Both Streptococcus and Actinomyces can produce acids from dietary sugars and are frequently found in caries lesions. In the oral cavity, nitrogenous compounds, such as peptides and amino acids, are provided continuously by saliva and crevicular gingival fluid. Given that these bacteria can also utilize nitrogen compounds for their growth, it was hypothesized that nitrogenous compounds may influence their acid production; however, no previous studies have examined this topic. Therefore, the present study aimed to assess the effects of nitrogenous compounds (tryptone and glutamate) on glucose‐derived acid production by Streptococcus and Actinomyces. Acid production was evaluated using a pH‐stat method under anaerobic conditions, whereas the amounts of metabolic end‐products were quantified using high performance liquid chromatography. Tryptone enhanced glucose‐derived acid production by up to 2.68‐fold, whereas glutamate enhanced Streptococcus species only. However, neither tryptone nor glutamate altered the end‐product profiles, indicating that the nitrogenous compounds stimulate the whole metabolic pathways involving in acid production from glucose, but are not actively metabolized, nor do they alter metabolic pathways. These results suggest that nitrogenous compounds in the oral cavity promote acid production by Streptococcus and Actinomyces in vivo.     

Temperature and diet affect carbon and nitrogen isotopes of fish muscle: can amino acid nitrogen isotopes explain effects?

Stable isotope ratios of carbon (δ¹³C) and nitrogen (δ¹⁵N) are widely used in food-web studies to determine trophic positioning and diet sources. However in order to accurately interpret stable isotope data the effects of environmental variability and dietary composition on isotopic discrimination factors and tissue turnover rates must be validated. We tested the effects of temperature and diet on tissue turnover rates and discrimination of carbon and nitrogen isotopes in an omnivorous fish, black bream (Acanthopagrus butcheri). Fish were raised at 16 °C or 23 °C and fed either a fish-meal or vegetable feed to determine turnover rates in fish muscle tissue up to 42 days after exposure to experimental treatments. Temperature and diet affected bulk tissue δ¹⁵N turnover and discrimination factors, with increased turnover and smaller discrimination factors at warmer temperatures. Fish reared on the vegetable feed showed greater bulk tissue δ¹⁵N changes and larger discrimination factors than those reared on a fish-meal feed. Temperature and diet affected bulk tissue δ¹³C values, however the direction of effects among treatments changed. Analyses of δ¹⁵N values of individual amino acids found few significant changes over time or treatment effects, as there was large variation at the individual fish level. However glutamic acid, aspartic acid and leucine changed most over the experiment and results mirrored those of treatment effects in bulk δ¹⁵N tissue values. The results demonstrate that trophic discrimination for δ¹⁵N and δ¹³C can be significantly different than those typically used in food-web analyses, and effects of diet composition and temperature can be significant. Precision of compound-specific isotope analyses (0.9‰) was larger than our effect size for bulk δ¹⁵N diet effects (0.7‰), therefore future experimental work in this area will need to establish a large effect size in order to detect significant differences. Our results also suggest that compound-specific amino acid δ¹⁵N may be useful for determining essential and non-essential amino acids for different animals.     

Intra‐lake stable isotope ratio variation in selected fish species and their possible carbon sources in Lake Kyoga (Uganda): implications for aquatic food web studies

The stable isotopes of nitrogen (δ¹⁵N) and carbon (δ¹³C) provide powerful tools for quantifying trophic relationships and carbon flow to consumers in food webs; however, the isotopic signatures of organisms vary within a lake. Assessment of carbon and nitrogen isotopic signatures in a suite of plants, invertebrates, and fishes in Lake Kyoga, indicated significant variation between two sites for δ¹³C (paired t = 6.305; df = 14, P < 0.001 and δ¹⁵N paired t = 1.292; df = 14; P < 0.05). The fish fauna in Bukungu was generally more ¹³C enriched (mean δ¹³C = –16.37 ± 1.64‰) than in Iyingo (mean δ¹³C = –20.80 ± 2.41‰) but more δ¹⁵N depleted (mean δ¹⁵N = 5.57 ± 0.71‰) than in Iyingo (mean δ¹⁵N = 6.92 ± 0.83‰). The simultaneous shifts in phytoplankton and consumer signatures confirmed phytoplankton as the major source of carbon for the food chain leading to fish. Limited sampling coverage within lakes may affect lake wide stable isotope signatures, and the same error is transferred into trophic position estimation. Consideration of potential intra‐lake spatial variability in isotope ratios and size is essential in evaluating the spatial and trophic structure of fish assemblages.     

Stable isotope records of sei whale baleens from Chilean Patagonia as archives for feeding and migration behavior

Carbon (δ¹³C) and nitrogen (δ¹⁵N) stable isotope variations in baleen plates of sei whales (Balaenoptera borealis) stranded after a mass mortality event in Chilean Patagonia were investigated to assess potential dietary and migratory patterns. Carbon and nitrogen isotope ratios of seven baleens from six individuals were analyzed. The δ¹³C values ranged from ? 19.1 to ? 15.9‰ and the δ¹⁵N values from 8.7 to 15.4‰. Variations of up to 2.9‰ for δ¹³C and 5.3‰ for δ¹⁵N were observed within one baleen. Carbon and nitrogen isotope records of each baleen were significantly correlated and showed recurring oscillations confirmed by wavelet analyses. Oscillations slightly differed in periodicity indicating variable baleen growth rates between 10.0 and 16.5 cm/year. Food sources of the whales are discussed in terms of available isotope data for potential prey taxa and potential migratory behavior on the basis of latitudinal isotope gradients of particulate organic matter. Cyclicity could be explained by regular migrations of the sei whales from subtropical calving areas to high‐latitude foraging grounds. δ¹⁵N records of baleens differed between individuals eventually pointing to diverse feeding and migratory preferences among sei whale individuals.     

Using stable isotopes to understand changes in ringed seal foraging ecology as a response to a warming environment

Trends toward increased temperatures, reduced sea ice extent, and longer open water seasons have resulted in changing Arctic ecosystem dynamics. Expected changes include shifts in distribution and abundance of prey species for seabirds and marine mammals. Using stable isotope analysis, we studied spatial and interannual variation in ringed seal (Pusa hispida) feeding ecology in Hudson Bay in relation to environmental variables, between 2003 and 2010. Ringed seal muscle and hair samples collected from Arviat and Sanikiluaq, Nunavut, were analyzed for stable isotope ratios of nitrogen (δ¹⁵N) and carbon (δ¹³C). Seals from western Hudson Bay (Arviat) had higher δ¹⁵N and lower δ¹³C than seals from eastern Hudson Bay (Sanikiluaq), and stable isotope ratios varied interannually within each region. Peak δ¹⁵N occurred in years with spring air temperatures between approximately ?5°C and ?2°C. This temperature range was characteristic of warm years in western Hudson Bay and cool years in eastern Hudson Bay. We hypothesize that the high δ¹⁵N observed in ringed seals is indicative of greater importance of capelin (Mallotus villosus) in ringed seal diet. A comparison of ringed seal isotopic niche widths indicated greater dietary differences between western and eastern Hudson Bay with warming, suggesting a possible ecological divergence related to climate change.     

Within-lake variability in carbon and nitrogen stable isotope signatures

1. We assessed spatial and temporal variation in carbon and nitrogen isotopic signatures in different compartments of a single lake ecosystem. Stable isotope analyses were made on samples of particulate organic matter (POM), zooplankton, periphyton, macrophytes, macroinvertebrates and fish collected from several locations throughout the ice‐free period. 2. No spatial variation in δ¹³C or δ¹⁵N values was found for pelagic samples of POM and zooplankton. However, pelagic δ¹⁵N signatures increased steadily through the summer resulting in an almost 6‰ average increase in POM and zooplankton. A concurrent decrease in epilimnetic nitrate concentrations suggested that the increase in δ¹⁵N of POM and zooplankton could have resulted from a progressive ¹⁵N‐enrichment of the available inorganic nitrogen pool as the size of this pool was reduced. 3. Significant spatial variation in isotopic ratios was observed within littoral and profundal communities. Some spatial differences were likely related to lake‐specific characteristics, such as a major inlet and a small harbour area and some were interconnected with temporal events. 4. Marked differences between spring and autumn δ¹⁵N and δ¹³C values of fish at one site probably reflected a spring spawning immigration from a larger downstream lake and also indicated limited dispersal of these immigrants. 5. Our results indicate that restricted sampling of ecosystem components from lakes may provide misleading single values for the isotope end members needed for quantitative uses of stable isotopes in mixing models and for estimating trophic position. Hence we strongly advise that studies of individual lakes, or multiple lake comparisons, that utilise stable isotope analyses should pay more attention to potential within lake spatial and temporal variability of isotope ratios.     

Differences in δ15N and δ13C between embryonic and maternal tissues of the ovoviviparous bluntnose sixgill shark Hexanchus griseus

Stable nitrogen (δ¹⁵N) and carbon (δ¹³C) isotope ratios from muscle, liver and yolk were analysed from the mother and embryos of an ovoviviparous shark, Hexanchus griseus. Embryonic liver and muscle had similar δ¹⁵N and δ¹³C ratios or were depleted in heavy isotopes, compared to the same maternal somatic and reproductive yolk tissues, but no relationship existed between δ¹⁵N or δ¹³C and embryo length, as expected, because a switch to placental nourishment is lacking in this species. This study expands the understanding of maternal nourishment and embryonic stable isotope differences in ovoviviparous sharks.