Stable Isotope Ratios as a Tool for Assessing Changes in Carbon and Nutrient Sources in Antarctic Terrestrial Ecosystems

Samples of an angiosperm species, nine lichen species and a terrestrial alga, were collected from a variety of Antarctic terrestrial habitats, and were analysed for C and N stable isotope composition. Collections were made along natural gradients, the marine gradient, running from the sea coast inland and the moisture gradient, determined by melt water and precipitation runoff, and running towards the sea coast. Considerable variation in stable isotope ratios was found; δ¹³C values ranged between −16 and −32‰ and δ¹⁵N values between −23 and +23‰ The variation in stable carbon isotope ratios could be attributed in part to species specific differences, but differences in water availability also played a role, as was shown for the terrestrial alga Prasiola crispa and the lichen species Usnea antarctica. The differences in the isotope ratios of nitrogen could be retraced to the origin of nitrogen: marine or terrestrial. The nitrogen stable isotope ratios were influenced by both the marine gradient from the sea inland and the melt water and precipitation flow running in the opposite direction, towards the sea. This was shown for the lichen species Turgidosculum complicatulum and the angiosperm species Deschampsia antarctica. The variation in the C and N stable isotope ratios can be used to determine sources and pathways of N and changes in the water availability in Antarctic terrestrial ecosystems. Contrary to earlier reports the use of stable N isotope ratios is possible in this case because of the relative simplicity of the structure of the Antarctic terrestrial ecosystems.     

Sheep Grazing Decreases Organic Carbon and Nitrogen Pools in the Patagonian Steppe: Combination of Direct and Indirect Effects

We explored the net effects of grazing on soil C and N pools in a Patagonian shrub–grass steppe (temperate South America). Net effects result from the combination of direct impacts of grazing on biogeochemical characteristics of microsites with indirect effects on relative cover of vegetated and unvegetated microsites. Within five independent areas, we sampled surface soils in sites subjected to three grazing intensities: (1) ungrazed sites inside grazing exclosures, (2) moderately grazed sites adjacent to them, and (3) intensely grazed sites within the same paddock. Grazing significantly reduced soil C and N pools, although this pattern was clearest in intensely grazed sites. This net effect was due to the combination of a direct reduction of soil N content in bare soil patches, and indirect effects mediated by the increase of the cover of bare soil microsites, with lower C and N content than either grass or shrub microsites. This increase in bare soil cover was accompanied by a reduction in cover of preferred grass species and standing dead material. Finally, stable isotope signatures varied significantly among grazed and ungrazed sites, with δ¹⁵N and δ¹³C significantly depleted in intensely grazed sites, suggesting reduced mineralization with increased grazing intensity. In the Patagonian steppe, grazing appears to exert a negative effect on soil C and N cycles; sound management practices must incorporate the importance of species shifts within life form, and the critical role of standing dead material in maintaining soil C and N stocks and biogeochemical processes. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Author Contributions  RAG designed study, performed research, analyzed data, wrote the paper; ATA designed study, wrote the paper; CGGM designed study, performed research, analyzed data; MGP performed research; OES designed study; RBJ designed study, contributed new methods.     

The Influence of Nutrient Availability on Soil Organic Matter Turnover Estimated by Incubations and Radiocarbon Modeling

We investigated the decomposability of soil organic matter (SOM) along a chronosequence of rainforest sites in Hawaii that form a natural fertility gradient and at two long-term fertilization experiments. To estimate turnover times and pool sizes of organic matter, we used two independent methods: (1) long-term incubations and (2) a three-box soil model constrained by radiocarbon measurements. Turnover times of slow-pool SOM (the intermediate pool between active and passive pools) calculated from incubations ranged from 6 to 20 y in the O horizon and were roughly half as fast in the A horizon. The radiocarbon-based model yielded a similar pattern but slower turnover times. The calculation of the ¹⁴C turnover times is sensitive to the lag time between photosynthesis and incorporation of organic C into SOM in a given horizon. By either method, turnover times at the different sites varied two- or threefold in soils with the same climate and vegetation community. Turnover times were fastest at the sites of highest soil fertility and were correlated with litter decay rates and primary productivity. However, experimental fertilization at the two least-fertile sites had only a small and inconsistent effect on turnover, with N slowing turnover and P slightly speeding it at one site. These results support studies of litter decomposition in suggesting that while plant productivity can respond rapidly to nutrient additions, decomposition may respond much more slowly to added nutrients.     

Stable isotope analyses provide new insights into ecological plasticity in a mixohaline population of European eel

Recent studies have shown that anguillid eel populations in habitats spanning the marine–freshwater ecotone can display extreme plasticity in the range of catadromy expressed by individual fishes. Carbon and nitrogen stable isotope analysis was used to differentiate between European eels (Anguilla anguilla) collected along a short (2 km) salinity gradient ranging from <1‰ to ~30‰ in Lough Ahalia, a tidal Atlantic lake system. Significant differences were recorded in mean δ¹³C, δ¹⁵N and C:N values from eels collected from fresh, brackish and marine-dominated basins. A discriminant analysis using these predictor variables correctly classified ca. 85% of eels to salinity zone, allowing eels to be classified as freshwater (FW), brackish (BW) or marine (MW) residents. The results of the discriminant analysis also suggested that a significant proportion of eels moved between habitats (especially between FW and BW). Comparisons of several key population parameters showed significant variation between eels resident in different salinity zones. Mean condition and estimated age was significantly lower in MW eels, whilst observed length at age (a correlate of growth) was significantly higher in MW eels, intermediate in BW and lowest in FW eels. This study has demonstrated that the ecology of eels found along a short salinity gradient can be extremely plastic and that stable isotope analysis has considerable utility in demonstrating intra-population variation in diadromous fishes.     

Soil respiration in northern forests exposed to elevated atmospheric carbon dioxide and ozone

The aspen free-air CO₂ and O₃ enrichment (FACTS II–FACE) study in Rhinelander, Wisconsin, USA, is designed to understand the mechanisms by which young northern deciduous forest ecosystems respond to elevated atmospheric carbon dioxide (CO₂) and elevated tropospheric ozone (O₃) in a replicated, factorial, field experiment. Soil respiration is the second largest flux of carbon (C) in these ecosystems, and the objective of this study was to understand how soil respiration responded to the experimental treatments as these fast-growing stands of pure aspen and birch + aspen approached maximum leaf area. Rates of soil respiration were typically lowest in the elevated O₃ treatment. Elevated CO₂ significantly stimulated soil respiration (8–26%) compared to the control treatment in both community types over all three growing seasons. In years 6–7 of the experiment, the greatest rates of soil respiration occurred in the interaction treatment (CO₂ + O₃), and rates of soil respiration were 15–25% greater in this treatment than in the elevated CO₂ treatment, depending on year and community type. Two of the treatments, elevated CO₂ and elevated CO₂ + O₃, were fumigated with ¹³C-depleted CO₂, and in these two treatments we used standard isotope mixing models to understand the proportions of new and old C in soil respiration. During the peak of the growing season, C fixed since the initiation of the experiment in 1998 (new C) accounted for 60–80% of total soil respiration. The isotope measurements independently confirmed that more new C was respired from the interaction treatment compared to the elevated CO₂ treatment. A period of low soil moisture late in the 2003 growing season resulted in soil respiration with an isotopic signature 4–6‰ enriched in ¹³C compared to sample dates when the percentage soil moisture was higher. In 2004, an extended period of low soil moisture during August and early September, punctuated by a significant rainfall event, resulted in soil respiration that was temporarily 4–6‰ more depleted in ¹³C. Up to 50% of the Earth’s forests will see elevated concentrations of both CO₂ and O₃ in the coming decades and these interacting atmospheric trace gases stimulated soil respiration in this study.     

Grazing and Ecosystem Carbon Storage in the North American Great Plains

Isotopic signatures of ¹³C were used to quantify the relative contributions of C₃ and C₄ plants to whole-ecosystem C storage (soil+plant) in grazed and ungrazed sites at three distinct locations (short-, mid- and tallgrass communities) along an east–west environmental gradient in the North American Great Plains. Functional group composition of plant communities, the source and magnitude of carbon inputs, and total ecosystem carbon storage displayed inconsistent responses to long-term livestock grazing along this gradient. C₄ plants [primarily Bouteloua gracilis (H.B.K.) Lag ex Steud.] dominated the long-term grazed site in the shortgrass community, whereas the ungrazed site was co-dominated by C₃ and C₄ species; functional group composition did not differ between grazed and ungrazed sites in the mid- and tallgrass communities. Above-ground biomass was lower, but the relative proportion of fine root biomass was greater, in grazed compared to ungrazed sites at all three locations. The grazed site of the shortgrass community had 24% more whole-ecosystem carbon storage compared to the ungrazed site (4022 vs. 3236 g C m⁻²). In contrast, grazed sites at the mid- and tallgrass communities had slightly lower (8%) whole-ecosystem carbon storage compared to ungrazed sites (midgrass: 7970 vs. 8683 g C m⁻²; tallgrass: 8273 vs. 8997 g C m⁻²). Differential responses between the shortgrass and the mid- and tallgrass communities with respect to grazing and whole-ecosystem carbon storage are likely a result of: (1) maintenance of larger soil organic carbon (SOC) pools in the mid- and tallgrass communities (7476–8280 g C m⁻²) than the shortgrass community (2517–3307 g C m⁻²) that could potentially buffer ecosystem carbon fluxes, (2) lower root carbon/soil carbon ratios in the mid- and tallgrass communities (0.06–0.10) compared to the shortgrass community (0.20–0.27) suggesting that variation in root organic matter inputs would have relatively smaller effects on the size of the SOC pool, and (3) the absence of grazing-induced variation in the relative proportion of C₃ and C₄ functional groups in the mid- and tallgrass communities. We hypothesize that the magnitude and proportion of fine root mass within the upper soil profile is a principal driver mediating the effect of community composition on the biogeochemistry of these grassland ecosystems.     

Intraguild interactions between spiders and ants and top-down control in a grassland food web

In most terrestrial ecosystems ants (Formicidae) as eusocial insects and spiders (Araneida) as solitary trappers and hunters are key predators. To study the role of predation by these generalist predators in a dry grassland, we manipulated densities of ants and spiders (natural and low density) in a two-factorial field experiment using fenced plots. The experiment revealed strong intraguild interactions between ants and spiders. Higher densities of ants negatively affected the abundance and biomass of web-building spiders. The density of Linyphiidae was threefold higher in plots without ant colonies. The abundance of Formica cunicularia workers was significantly higher in spider-removal plots. Also, population size of springtails (Collembola) was negatively affected by the presence of wandering spiders. Ants reduced the density of Lepidoptera larvae. In contrast, the abundance of coccids (Ortheziidae) was positively correlated with densities of ants. To gain a better understanding of the position of spiders, ants and other dominant invertebrate groups in the studied food web and important trophic links, we used a stable isotope analysis (¹⁵N and ¹³C). Adult wandering spiders were more enriched in ¹⁵N relative to ¹⁴N than juveniles, indicating a shift to predatory prey groups. Juvenile wandering and web-building spiders showed δ¹⁵N ratios just one trophic level above those of Collembola, and they had similar δ¹³C values, indicating that Collembola are an important prey group for ground living spiders. The effects of spiders demonstrated in the field experiment support this result. We conclude that the food resource of spiders in our study system is largely based on the detrital food web and that their effects on herbivores are weak. The effects of ants are not clear-cut and include predation as well as mutualism with herbivores. Within this diverse predator guild, intraguild interactions are important structuring forces.     

Resolving temporal variation in vertebrate diets using naturally occurring stable isotopes

Assessments of temporal variation in diets are important for our understanding of the ecology of many vertebrates. Ratios of naturally occurring stable isotopes in animal tissues are a combination of the source elements and tissue specific fractionation processes, and can thus reveal dietary information. We review three different approaches that have been used to resolve temporal diet variation through analysis of stable isotopes. The most straightforward approach is to compare samples from the same type of tissue that has been sampled over time. This approach is suited to address either long or short-term dietary variation, depending on sample regime and which tissue that is sampled. Second, one can compare tissues with different metabolic rates. Since the elements in a given tissue have been assimilating during time spans specific to its metabolic rate, tissues with different metabolic rates will reflect dietary records over different periods. Third, comparisons of sections from tissues with progressive growth, such as hair, feathers, claws and teeth, will reveal temporal variation since these tissues will retain isotopic values in a chronological order. These latter two approaches are mainly suited to address questions regarding intermediate and short-term dietary variation. Knowledge of tissue specific metabolic rates, which determine the molecular turnover for a specific tissue, is of central importance for all these comparisons. Estimates of isotopic fractionation between source and measured target are important if specific hypotheses regarding the source elements are addressed. Estimates of isotopic fractionation, or at least of differences in fractionation between tissues, are necessary if different tissues are compared. We urge for more laboratory experiments aimed at improving our understanding of differential assimilation of dietary components, isotopic fractionation and metabolic routing. We further encourage more studies on reptiles and amphibians, and generally more studies utilizing multiple tissues with different turnover rates.     

Flight capabilities and feeding habits of silphine beetles: are flightless species really “carrion beetles”?

We determined the flight capabilities and feeding habits of adults of nine silphine beetle species and illustrated their relationship. We examined the silphine beetles for the presence or absence of flight muscles and estimated their feeding habits by comparing the carbon and nitrogen stable isotope ratios for them with those of necrophagous nicrophorine species and carnivorous carabine species. Three species (Silpha longicornis, S. perforata and Phosphuga atrata) completely lacked individuals with flight muscles, and one species (Eusilpha japonica) showed flight muscle dimorphism. Stable isotope analysis suggested that these species were carnivores, mainly feeding on soil invertebrates. Most flight species showed higher isotopic ratios than the flightless species. Some of them have isotopic ratios close to those of the nicrophorine species, suggesting that these species mainly feed on vertebrate carcasses. Flightless silphine species would have limited ability to search for patchy and unpredictable carcass resources. Further studies are necessary to understand the adaptive evolution of flight capability and the feeding habits in this group.     

Holocene palaeoenvironment in a former coastal lagoon of the arid south eastern Iberian Peninsula: salinization effects on δ<Superscript>15</Superscript>N

The palaeoenvironment of a former coastal lagoon in the south eastern Iberian Peninsula (San Rafael, Almeria, Spain) were inferred from one core analyzed for particulate organic matter content (POM) together with its C/N, δ¹³C, δ¹⁵N to depict the biogeochemical record from the Late Glacial to the Holocene. The results, complemented by previously reported pollen assemblages, indicate the appearance of a freshwater lagoon at 7300 b.p. (uncalibrated ¹⁴C age), its salinization at 6200 b.p. and its disappearance at 4400 b.p. The period of existence of the lagoon coincided with a period of wetter conditions as inferred from terrestrial vegetation. The lagoon’s salinization was not related to a decrease in precipitation but to a stronger maritime influence since there were no parallel changes in terrestrial vegetation. Salinization caused an increase in δ¹³C, associated with a higher relative presence of C₄ plants, and an increase in δ¹⁵N, due to a decrease in plant N demand. The late period of the lagoon, from about 5100 to 4400 b.p., shows a progressive drying and salinization not detected in isotopes but reflected in a decrease in POM, and in the pollen records. Increases in δ¹⁵N were related to increases in salinity within the lagoon, and are indicative of a more open N cycle, because the absence of changes in terrestrial vegetation rules out changes in the catchment area as the cause for changes in δ¹⁵N.     

Stable Isotopic and Trace Elemental Compositions of Otoliths and the Stock Structure of Pacific Cod, Gadus macrocephalus

Synopsis We examined the stock composition and life history of Pacific cod, Gadus macrocephalus, in the northeast Pacific Ocean from sagittal otoliths collected from three marine fishing areas in Washington State, U.S.A. We analyzed both stable isotope ratios (δ¹⁸O and δ¹³C) and trace elemental concentrations (Sr, Mg, Na, Fe, Mn) for these otoliths. The combination of δ¹⁸O and δ¹³C, and correlation of δ¹⁸O vs. 1000Sr/Ca and 1000Mg/Ca showed clear separations between North Puget Sound and coastal cod, suggesting there might be two different spawning stocks in the region. The North Puget Sound cod might represent an ‘Estuary-type’ from the Strait of Georgia, whereas coastal cod might represent an ‘Ocean-type’ from the Pacific west coast. Isotopic variations from five representative otoliths also showed a two-stage life history for Pacific cod, and a critical transition period for cod in migration to the ocean or in age of sexual maturity. These chemical interpretations and conclusions appear in agreement with biological observations of Pacific cod, and are consistent with results of previous studies for other marine fish species in nearby areas.     

Disentangling effects of growth and nutritional status on seabird stable isotope ratios

A growing number of studies suggest that an individual’s physiology affects its carbon and nitrogen stable isotope signatures, obscuring a signal often assumed to be only a reflection of diet and foraging location. We examined effects of growth and moderate food restriction on red blood cell (RBC) and feather δ¹⁵N and δ¹³C in rhinoceros auklet chicks (Cerorhinca monocerata), a piscivorous seabird. Chicks were reared in captivity and fed either control (75 g/day; n = 7) or ~40% restricted (40 g/day; n = 6) amounts of high quality forage fish. We quantified effects of growth on isotopic fractionation by comparing δ¹⁵N and δ¹³C in control chicks to those of captive, non-growing subadult auklets (n = 11) fed the same diet. To estimate natural levels of isotopic variation, we also collected blood from a random sample of free-living rhinoceros auklet adults and chicks in the Gulf of Alaska (n = 15 for each), as well as adult feather samples (n = 13). In the captive experiment, moderate food restriction caused significant depletion in δ¹⁵N of both RBCs and feathers in treatment chicks compared to control chicks. Growth also induced depletion in RBC δ¹⁵N, with chicks exhibiting lower δ¹⁵N when they were growing the fastest. As growth slowed, δ¹⁵N increased, resulting in an overall pattern of enrichment over the course of the nestling period. Combined effects of growth and restriction depleted δ¹⁵N in chick RBCs by 0.92‰. We propose that increased nitrogen-use efficiency is responsible for ¹⁵N depletion in both growing and food-restricted chicks. δ¹⁵N values in RBCs of free-ranging auklets fell within a range of only 1.03‰, while feather δ¹⁵N varied widely. Together, our captive and field results suggest that both growth and moderate food restriction can affect stable isotope ratios in an ecologically meaningful way in RBCs although not feathers due to greater natural variability in this tissue.     

Isotopic signatures and nutrient relations of plants inhabiting brackish wetlands in the northeastern coastal plain of Venezuela

The semi-diurnal tidal regime (≥2 m) in the Paria Gulf on the Atlantic coast of Venezuela, and the flat landscape of the region, allow the penetration for tens of km of marine waters into the rivers draining the northeastern coastal plain of the country. The levels of salinity, tidal flooding, and sedimentation decrease perpendicularly from the river channel toward the back swamps. The vegetation varies sequentially from fringe mangroves along the river margins, to back swamps containing forests dominated by Pterocarpus officinalis, herbaceous communities of Lagenocarpus guianensis, and palm swamps with Mauritia flexuosa, Chrysobalanus icaco, and Tabebuia spp. This environmental structure was used to test the hypotheses that: (a) mangrove distribution is strongly associated with salinity of interstitial water, and (b) they occupy areas where tidal influence and sediment dynamics determine a relatively open N cycle. Analyses of soil, water, and plants along a 1.5 km transect located near the confluence of the Guanoco and San Juan Rivers (Sucre and Monagas States, Venezuela) revealed that: (a) conductivity decreased from 11 to 0.2 mmhos cm⁻¹ from the river fringe to the internal swamp, whereas Na in the same stretch decreased from 100 to 2 μM; (b) average leaf tissue concentrations of Na, P, and N decreased significantly along the transect; (c) P. officinalis showed a large Na-exclusion capacity indicated by positive K/Na ratios from 8 to 200, and Crinum erubescens counteracted Na by accumulating K above 1,000 mmol kg⁻¹; (d) leaves varied widely in δ ¹³C (−25.5 to −32‰) and δ ¹⁵N (4 to −10.5‰) values. Samples were aggregated according to soil carbon content corresponding to those of the mangrove forest belt (5–28 mol C kg⁻¹; 0–650 from river fringe) and those of the back swamps (40–44 mol C kg⁻¹; 700–1,500 m from river fringe). The concentrations of Na, P, and N (in mmol kg⁻¹) and δ ¹⁵N values (in ‰) were significantly higher in the mangrove forest compared to the back swamp (Na 213 vs. 88; P 41 vs. 16; N 1,535 vs. 727; δ ¹⁵N 1.5 vs. −3.7), indicating that the fringe forest was not nutrient limited. These results support the hypotheses that mangroves are restricted to the more-saline sections of the transect, and that the fringe forest has a more open N cycle, favoring ¹⁵N accumulation within the system.     

Stable isotope analyses of otoliths in identification of hatchery origin of Atlantic salmon (Salmo salar) in Maine

We conducted stable oxygen and carbon isotope analyses for otoliths of Atlantic salmon (Salmo salar), in an attempt to develop a reference database on isotopic variability among private and federal hatcheries in Maine which currently support the salmon aquaculture industry and recovery of endangered populations. During the first phase of our study, we collected 40–50 sagittal otoliths of juvenile Atlantic salmon from each of the five hatcheries and analyzed for stable oxygen and carbon isotope ratios (¹⁸O/¹⁶O or δ¹⁸O, and ¹³C/¹²C or δ¹³C). Combination of δ¹⁸O and δ¹³C signatures in otoliths showed that the five hatcheries can be clearly separated and chemically distinguished. By identifying stable isotopic variations of otoliths from different hatchery settings, we were able to establish some isotopic criteria or standards to assign a likelihood that an individual Atlantic salmon came from a specific hatchery within the reference database. If successful, a diagnostic tool that can provide definitive information on identification of the hatchery origin could serve as a novel marking technique, and the chemical method may provide a more effective alternative to DNA analysis for mixed stocks. Overall our isotopic data from otoliths support the hypothesis that there are detectable differences between the five hatcheries, and multiple statistical analyses indicated that we can correctly distinguish individual Atlantic salmon into a hatchery with high confidence.     

Trophic Niche Segregation in the Nilotic Ichthyofauna of Lake Albert (Uganda, Africa)

Synopsis Nile perch, Lates niloticus, and Nile tilapia, Oreochromis niloticus, were originally transplanted from Lake Albert in western Uganda to the African Great Lakes, Lake Victoria and Lake Kyoga, where they are partially implicated in reduction of the fish species diversity. Lake Albert is facing multiple environmental changes, including declining fish species diversity, hyper-eutrophication, hypoxia, and reduced fish catches. To examine the role of Nile perch and Nile tilapia in the food web in their native Lake Albert, we estimated their diets using stable nitrogen and carbon isotopes. In Lake Albert, the tilapiine congeners (closely related species), Tilapia zillii, Oreochromis leucostictus, and Sarethorodon galilaeus, and the centropomid Nile perch congener, Lates macrophthalmus, have narrower diet breath in the presence of the native O. niloticus and L. niloticus. A computerized parameter search of dietary items for five commercially important fish species (Hydrocynus forskahlii, Bagrus bayad, L. niloticus, Alestes baremose and Brycinus nurse) was completed using a static isotopic mixing model. The outcome of the simulation for most fish species compared favorably to previously published stomach contents data for the Lake Albert fishes dating back to 1928, demonstrating agreement between stable isotope values and analyses of stomach contents. While there were some indications of changes in the diets of L. niloticus and A. baremose diets over the past 20 years in parallel with other changes in the lake, for the most part, food web structure in this lake remained stable since 1928. The Lake Albert fish assemblage provides insight into the invasion success of L. niloticus and O. niloticus.     

Temporal and spatial variability in stable isotope compositions of a freshwater mussel: implications for biomonitoring and ecological studies

Stable isotopes can be used to elucidate ecological relationships in community and trophic studies. Findings are calibrated against baselines, e.g. from a producer or primary consumer, assumed to act as a reference to the isotopic context created by spatio-temporal attributes such as geography, climate, nutrient, and energy sources. The ability of an organism to accurately represent a community base depends on how, and over what time-scale, it assimilates ambient materials. Freshwater mussels have served as references for trophic studies of freshwater communities and as indicators of change in nutrient pollution load or source. Their suitability as reference animals has not yet been fully explored, however. We conducted a series of studies examining the suitability of freshwater mussels as isotopic baselines, using their ability to reflect variation in ambient nutrient loads as a case scenario. (1) We analyzed bivalve foot tissue δ¹⁵N and δ¹³C from 22 stream reaches in the Piedmont region of North Carolina, USA to show that compositions varied substantially among locations. Site mean bivalve δ¹³C values correlated with site ambient particulate organic matter (POM) δ¹³C values, and site mean bivalve δ¹⁵N values correlated with site ambient water dissolved δ¹⁵N-NO₃ values. (2) Similarity of results among sample types demonstrated that the minimally invasive hemolymph sample is a suitable substitute for foot tissue in δ¹⁵N analyses, and that small sample sizes generate means representative of a larger population. Both findings can help minimize the impact of sampling on imperiled freshwater mussel populations. (3) In a bivalve transplantation study we showed that hemolymph δ¹⁵N compositions responded to a shift in ambient dissolved δ¹⁵N-NO₃, although slowly. The tissue turnover time for bivalve hemolymph was 113 days. We conclude that bivalves serve best as biomonitors of chronic, rather than acute, fluctuations in stream nutrient loads, and provide initial evidence of their suitability as time-integrated isotopic baselines for community studies.     

Nitrogen and carbon isotope responses of Chinese cabbage and chrysanthemum to the application of liquid pig manure

The effects of the liquid pig manure (LM) used in organic farming on the natural abundance of ¹⁵N and ¹³C signatures in plant tissues have not been studied. We hypothesized that application of LM will (1) increase δ¹⁵N of plant tissues due to the high δ¹⁵N of N in LM as compared with soil N or inorganic fertilizer N, and (2) increase δ¹³C of plant tissues as a result of high salt concentration in LM that decreases stomatal conductance of plants. To test these hypotheses, variations in the δ¹⁵N and δ¹³C of Chinese cabbage (Brassica campestris L.) and chrysanthemum (Chrysanthemum morifolium Ramatuelle) with two different LMs (with δ¹⁵N of +15.6 and +18.2‰) applied at two rates (323 and 646 kg N ha^-1 for cabbage and 150 and 300 kg N ha^-1 for chrysanthemum), or urea (δ¹⁵N = -2.7‰) applied at the lower rate above for the respective species, in addition to the control (no N input) were investigated through a 60-day pot experiment. Application of LM significantly increased plant tissue δ¹⁵N (range +9.4 to +14.9‰) over the urea (+3.2 to +3.3‰) or control (+6.8 to 7.7‰) treatments regardless of plant species, strongly reflecting the δ¹⁵N of the N source. Plant tissue δ¹³C were not affected by the treatments for cabbage (range −30.8 to −30.2‰) or chrysanthemum (−27.3 to −26.8‰). However, cabbage dry matter production decreased while its δ¹³C increased with increasing rate of LM application or increasing soil salinity (P < 0.05), suggesting that salinity stress caused by high rate of LM application likely decreased stomatal conductance and limited growth of cabbage. Our study expanded the use of the δ¹⁵N technique in N source (organic vs. synthetic fertilizer) identification and suggested that plant tissue δ¹³C maybe a sensitive indicator of plant response to salinity stress caused by high LM application rates.     

δ13C and δ15N changes after dietary shift in veliger larvae of the slipper limpet Crepidula fornicata: an experimental evidence

δ¹³C and δ¹⁵N measurements are still poorly conducted in benthic invertebrate larvae. To assess the δ¹³C and δ¹⁵N changes occurring after a dietary shift, experiments were conducted on veliger larvae of Crepidula fornicata fed with two cultured microalgae (Isochrysis galbana and Pavlova lutheri) of known isotopic composition, ¹³C-enriched and ¹⁵N-depleted compared to the initial values of the larvae. Rapid changes in larval δ¹³C and δ¹⁵N were observed after the dietary shift, with an increase in δ¹³C and a decrease in δ¹⁵N. After 19 days of feeding, isotopic equilibrium was still not reached, a period which is close to the duration of the pelagic life of the larvae. This implies that the isotopic composition measured in field-collected larvae might only partly reflect actual larval feeding but also the parental isotopic signature, especially during the early developmental stages. Isotopic measurements in marine invertebrate larvae should thus be interpreted cautiously. In planktonic food web investigations, the study of field-collected larvae of different size/developmental stage may reduce potential misinterpretations.     

Seasonal variation in photosynthesis in six woody species with different leaf phenology in a valley savanna in southwestern China

During 2003–2005, we examined the effect of seasonal drought on water status, gas exchange, δ¹³C, chlorophyll fluorescence and spectral reflectance in six woody species in a valley savanna near the Yuanjiang River (the upper Red River) in southwestern China. Three different phenological types of these woody species were compared, i.e., an evergreen species, Cyclobalanopsis helferiana, two winter-deciduous (WD) species, Buchanania latifolia and Symplocos racemosa, and three drought-deciduous (DD) species, Terminthia paniculata, Wendlandia tinctoria and Woodfordia fruticosa. We aimed to test the following three hypotheses: (1) the evergreen and WD species employ a drought avoidance strategy, whereas DD species employ a drought tolerance strategy; (2) the evergreen and WD species have a more economical water use strategy than the DD species and (3) the evergreen and WD species have a stronger photoprotection capacity through thermal dissipation than the DD species. At the end of a prolonged drought, the predawn leaf water potential (Ψ_pd) in C. helferiana and S. racemosa dropped to ca. −0.8 MPa, whereas the Ψ_pd in B. latifolia remained close to zero and DD species were leafless. In the rainy seasons, maximal photosynthetic rates of the evergreen (18.4 μmol m⁻² s⁻¹) and W. fruticosa (18.0 μmol m⁻² s⁻¹) were higher than those of the other four species (12.2−13.8 μmol m⁻² s⁻¹). The evergreen and WD species responded to drought by closing stomata and thus maintained a constant relative water content (RWC), which is a typical drought avoidance strategy; however, it is at the expense of carbon gain. DD species maintained a high photosynthetic capacity with a decrease in both stomatal conductance and RWC until the driest period, and then shifted from the drought tolerance strategy to the avoidance mechanism by shoot dieback. There was no significant difference in the means of δ¹³C across the phenological groups. The evergreen and WD species had stronger heat dissipation than the DD species in dry seasons. All species increased leaf spectral reflectance, probably because of degradation of chlorophyll as indicated by the leaf reflectance index, which should reduce light harvesting. All species showed a strong increase in the ratio of red to green spectral reflectance of leaves during dry seasons, indicating the accumulation of anthocyanin, which may contribute to screening sunlight and scavenging reactive oxygen species. Different responses to drought of savanna woody species with different leaf phenologies may facilitate the partitioning of resource use and hence their co-existence.     

The effect of cold-induced increased metabolic rate on the rate of <Superscript>13</Superscript>C and <Superscript>15</Superscript>N incorporation in house sparrows (<Emphasis Type="Italic">Passer domesticus</Emphasis>)

Animals with high metabolic rates are believed to have high rates of carbon and nitrogen isotopic incorporation. We hypothesized that (1) chronic exposure to cold, and hence an increase in metabolic rate, would increase the rate of isotopic incorporation of both ¹³C and ¹⁵N into red blood cells; and (2) that the rate of isotopic incorporation into red blood cells would be allometrically related to body mass. Two groups of sparrows were chronically exposed to either 5 or 22°C and switched from a ¹³C-depleted C₃-plant diet to a more ¹³C-enriched C₄-plant one. We used respirometry to estimate the resting metabolic rate of birds exposed chronically to our two experimental temperatures. The allometric relationship between the rate of ¹³C incorporation into blood and body mass was determined from published data. The of birds at 5°C was 1.9 times higher than that of birds at 22°C. Chronic exposure to a low temperature did not have an effect on the rate of isotopic incorporation of ¹⁵N save for a very small effect on the incorporation of ¹³C. The isotopic incorporation rate of ¹³C was 1.5 times faster than that of ¹⁵N. The fractional rate of ¹³C incorporation into avian blood was allometrically related to body mass with an exponent similar to −1/4. We conclude that the relationship between metabolic rate and the rate of isotopic incorporation into an animal’s tissues is indirect. It is probably mediated by protein turnover and thus more complex than previous studies have assumed.