Carbon and nitrogen cycling in a lead polluted grassland evaluated using stable isotopes (δ13C and δ15N) and microbial,plant and soil parameters

Plant and Soil - Carbon (C) and nitrogen (N) cycling are key ecosystem functions potentially altered by heavy metal pollution. We used an ecosystem approach to study the long-term effect of lead...     

Tree population changes in a tropical dry evergreen forest of south India over a decade (1992–2002)

Changes in species composition and density of trees >10 cm gbh in a tropical dry evergreen forest in Puthupet, south India are interpreted for the period between 1992 and 2002. A 1-ha plot was inventoried in 1992 and was recensused in 2002. During the 10-year interval tree taxa diversity as well as stand density increased, but the basal area value decreased. Tree species richness increased by 21% (from 24 to 29 species) by an addition of eight species and local extinction of three species. The tree density increased just by eight individuals (from 1330 stems ha⁻¹ in 1992 to 1338 ha⁻¹ in 2002), but the basal area decreased by 8% (from 37.5 to 34.5 m² ha⁻¹). Many species (11 numbers) have increased in abundance rather than decreased. Many surviving species seem to have considerable stability in abundance at the local scale. The density of smaller stems (10 29 cm gbh) increased by 15.3%, while that of the larger trees decreased drastically (81.6%). Ninety percent of the missing stems were from the middlestorey of the forest. Tree density changes among the three ecological guilds revealed a decrease in stem density and an increase in basal area in the lowerstorey; while the middlestorey exhibited a reverse trend. Family-wise, tree density changes revealed that the majority of families (67%) showed an increase in stem density. Long-term studies on tree population changes are essential to estimate tree mortality and recruitment rates, which will provide a greater insight in tropical forest dynamics.     

δ13C and δ15N indicators of fish and shrimp community diet and trophic structure in a mangrove ecosystem in Thailand

Stable carbon and nitrogen isotope ratios were used to elucidate primary carbon sources and trophic relationships of the fish and shrimp community in the Klong Ngao mangrove ecosystem, southern Thailand. There were no significant differences in isotopic compositions of biota between mangrove and offshore sites (Welch–Aspin test). The δ¹⁵N values of eight fish species and two shrimp species at both sites were also not significantly different by the test, meaning that at both sites they feed on the same diets due to the discharge of large quantities of mangrove sediments. The δ¹⁵N isotopic enrichment of consumers suggested that there are four trophic levels in the Klong Ngao food web, with at least two fish species capable of switching feeding strategies and thus altering their apparent trophic positions. Phytoplankton culture experiments indicated that mangrove-derived sediments could play an important role in stimulating phytoplankton growth for low turbidity offshore areas, thus providing an alternate food source. The isotopic associations among sources and consumers indicated that mangroves were the major carbon source supporting aquatic food webs in the Klong Ngao ecosystem.     

Research article: small-scale spatial variation of δ13C and δ15N isotopes in Antarctic carbon sources and consumers

Regional food web studies that fail to account for small-scale isotopic variability can lead to a mismatch between an organism’s inferred and true trophic position. Misinterpretation of trophic status may result, substantially limiting spatial and temporal comparability of food web studies. We sampled several carbon sources and consumers in a nested design to assess the variability of food web members across small spatial scales (100 s of m to several km) in regions around the Windmill Islands and Vestfold Hills in East Antarctica. For carbon sources, δ¹³C in sea ice POM was particularly variable between locations (km apart) and between sites (100 s of m apart) with replicate samples varying by up to 16‰. Macroalgae δ¹³C was less variable (replicate samples ranging up to 6.9‰ for the red alga Iridaea cordata), yet still differed between locations. Sediment POM and pelagic POM were the least variable, displaying minimal differences between locations or sites for δ¹³C and δ¹⁵N. Three out of eight consumers were significantly different between locations for δ¹³C, and five out of eight for δ¹⁵N, with the fish Trematomus bernacchii the most variable for both δ¹³C and δ¹⁵N. At smaller scales, the amphipod Paramorea walkeri showed significant variation between sites in δ¹³C but not in δ¹⁵N. We attribute small-scale variability to the dynamic physical environment for carbon sources in coastal systems and a close coupling of diet to habitat for consumers. We highlight the need to account for small-scale spatial variation in sampling designs for regional food web studies.     

Large 13C/12C and small 15N/14N isotope fractionation in an experimental detrital foodweb (litter–fungi–collembolans)

Correctly estimating the trophic fractionation factors (Δ¹⁵N and Δ¹³C) in controlled laboratory conditions is essential for the application of stable isotope analysis in studies on the trophic structure of soil communities. Laboratory experiments usually suggest large ¹⁵N/¹⁴N and small ¹³C/¹²C trophic fractionation, but in field studies litter-dwelling microarthropods and other invertebrates are consistently enriched in ¹³C relative to plant litter. In the present study, we report data from two laboratory experiments investigating both fungi–collembolans and litter–fungi–collembolans systems. In the fungi–collembolans system, Δ¹⁵N and Δ¹³C averaged 1.4 ± 0.1 and 1.0 ± 0.2 ‰, respectively. In microcosms with fungi-inoculated litter, the difference in δ¹⁵N between collembolans and plant litter averaged 1.5 ± 0.2 ‰, confirming the relatively small ¹⁵N/¹⁴N trophic fractionation at the basal level of detrital foodwebs reported in numerous field studies. In full agreement with field observations, the difference in δ¹³C between bulk litter and collembolans in laboratory microcosms averaged 3.6 ± 0.1 ‰ and only little depended on collembolan species identities or the presence of water-soluble compounds in the litter. We conclude that increased δ¹³C values typical of litter-dwelling decomposers are largely determined by an increased ¹³C content in saprotrophic microorganisms.     

Determining the appropriate pretreatment procedures and the utility of liver tissue for bulk stable isotope (δ13C and δ15N) studies in sharks

Stable-isotope analysis (SIA) provides a valuable tool to address complex questions pertaining to elasmobranch ecology. Liver, a metabolically active, high turnover tissue (~166 days for 95% turnover), has the potential to reveal novel insights into recent feeding/movement behaviours of this diverse group. To date, limited work has used this tissue, but ecological application of SIA in liver requires consideration of tissue preparation techniques given the potential for high concentrations of urea and lipid that could bias δ¹³C and δ¹⁵N values (i.e., result in artificially lower δ¹³C and δ¹⁵N values). Here we investigated the effectiveness of (a) deionized water washing (WW) for urea removal from liver tissue and (b) chloroform-methanol for extraction of lipids from this lipid rich tissue. We then (a) established C:N thresholds for deriving ecologically relevant liver isotopic values given complications of removing all lipid and (b) undertook a preliminary comparison of δ¹³C values between tissue pairs (muscle and liver) to test if observed isotopic differences correlated with known movement behaviour. Tests were conducted on four large shark species: the dusky (DUS, Carcharhinus obscurus), sand tiger (RAG, Carcharias taurus), scalloped hammerhead (SCA, Sphyrna lewini) and white shark (GRE, Carcharodon carcharias). There was no significant difference in δ¹⁵N values between lipid-extracted (LE) liver and lipid-extracted/water washed (WW) treatments, however, WW resulted in significant increases in %N, δ¹³C and %C. Following lipid extraction (repeated three times), some samples were still biased by lipids. Our species-specific “C:N thresholds” provide a method to derive ecologically viable isotope data given the complexities of this lipid rich tissue (C:N thresholds of 4.0, 3.6, 4.7 and 3.9 for DUS, RAG, SCA and GRE liver_LEWW tissue, respectively). The preliminary comparison of C:N threshold corrected liver and muscle δ¹³C values corresponded with movement/habitat behaviours for each shark; minor differences in δ¹³C values were observed for known regional movements of DUS and RAG (δ¹³C_Diffs = 0.24 ± 0.99‰ and 0.57 ± 0.38‰, respectively), while SCA and GRE showed greater differences (1.24 ± 0.63‰ and 1.08 ± 0.71‰, respectively) correlated to large-scale movements between temperate/tropical and pelagic/coastal environments. These data provide an approach for the successful application of liver δ¹³C and δ¹⁵N values to examine elasmobranch ecology.     

Effect of nutritional condition on variation in δ13C and δ15N stable isotope values in Pumpkinseed sunfish (Lepomis gibbosus) fed different diets

Stable isotope analysis is frequently used to infer resource use in natural populations of fishes. Studies have examined factors, other than diet, that influence δ¹⁵N and δ¹³C including tissue-specific rates of equilibration and starvation. Most such studies completed under laboratory conditions tightly control food quantity and its isotopic composition, but it is also necessary to evaluate the influence of these factors under more natural conditions. Using pumpkinseed sunfish (Lepomis gibbosus) we evaluated whether restricted rations below minimum daily requirements affects tissue equilibration to a change in diet by holding fish on two treatments that often reflect divergent resource use in natural populations (pelagic zooplankton or littoral macroinvertebrates). Over 42 days, δ¹⁵N values increased while δ¹³C values did not change, additionally neither were related to diet treatment. Increased δ¹⁵N values were negatively related to body condition while δ¹³C values were not, indicating that stable isotope values were more affected by decreasing body condition than by diet. Additionally, δ¹⁵N values changed more in the blood and liver tissues than in white muscle tissue, indicating that restricting food availability had greater effects on tissues with greater metabolic activity. We hypothesize that stable isotope values of consumers are subject to a tissue-specific trade-off between sensitivity to changes in resource use and resistance to the effects of low resource availability. This trade-off may require consideration in stable isotope studies of wild populations facing periodic limitations of food availability.     

Variation in hair δ13C and δ15N values in long-tailed macaques (Macaca fascicularis) from Singapore

Much of the primatology literature on stable isotope ratios of carbon (δ¹³C) and nitrogen (δ¹⁵N) has focused on African and New World species, with comparatively little research published on Asian primates. Here we present hair δ¹³C and δ¹⁵N isotope values for a sample of 33 long-tailed macaques from Singapore. We evaluate the suggestion by a previous researcher that forest degradation and biodiversity loss in Singapore have led to a decline in macaque trophic level. The results of our analysis indicated significant spatial variability in δ¹³C but not δ¹⁵N. The range of variation in δ¹³C was consistent with a diet based on C₃ resources, with one group exhibiting low values consistent with a closed canopy environment. Relative to other macaque species from Europe and Asia, the macaques from Singapore exhibited a low mean δ¹³C value but mid-range mean δ¹⁵N value. Previous research suggesting a decline in macaque trophic level is not supported by the results of our study.     

White spruce foliar δ13C and δ15N indicate changed soil N availability by understory removal and N fertilization in a 13-year-old boreal plantation

Background and Aims

Phosphorus (P) mineralisation from crop residues is usually predicted from total P or carbon: phosphorus (C: P) ratios. However, these measures have limited accuracy as they do not take into account the presence of different P species that may be mineralised at different rates. In this study P speciation was determined using solution ³¹P nuclear magnetic resonance (NMR) spectroscopy to understand the potential fate of residue P in soils.

Methods

Mature above-ground biomass of eight different crops sampled from the field was portioned into stem, chaff and seed.

Results

The main forms of P detected in stem and chaff were orthophosphate (25–75 %), phospholipids (10–40 %) and RNA (5–30 %). Phytate was the dominant P species in seeds, and constituted up to 45 % of total P in chaff but was only detected in minor amounts (<1 %) in stem residue. The majority (65–95 %) of P in stems was water-extractable, and most of this was detected as orthophosphate. However, this includes organic P that may have been hydrolysed during the water extraction.

Conclusions

This study indicates that the majority of residue P in aboveground plant residues has the potential to be delivered to soil in a form readily available to plants and soil microorganisms.

     

Bomb-pulse 14C analysis combined with 13C and 15N measurements in blood serum from residents of Malmö, Sweden

The ¹⁴C content of 60 human blood serum samples from residents of Malmö (Sweden) in 1978, obtained from a biobank, has been measured to estimate the accuracy of ¹⁴C bomb-pulse dating. The difference between the date estimated using the Calibomb software and sampling date varied between ?3 ± 0.4 and +0.2 ± 0.5 years. The average age deviation of all samples was ?1.5 ± 0.7 years, with the delay between production and consumption of foodstuffs being probably the dominating cause. The potential influence of food habits on the ¹⁴C date has been evaluated using stable isotope δ¹³C and δ¹⁵N analysis and information about the dietary habits of the investigated individuals. Although the group consisting of lacto-ovo vegetarians and vegans (pooled group) was not completely separated from the omnivores in a stable isotopic trophic level diagram, this analysis proved to add valuable information on probable dietary habits. The age deviation of the sampling date from the respective Calibomb date was found strongly correlated with the δ¹³C values, probably due to influence from marine diet components. For the omnivore individuals, there were indications of seasonal effects on δ¹³C and the age deviation. No significant correlation was found between the age deviation and the δ¹⁵N values of any dietary group. No influence of sex or year of birth was found on neither the ¹⁴C nor the δ¹³C and δ¹⁵N values of the serum samples. The data were also divided into two groups (omnivores and pooled group), based on the level of δ¹⁵N in the samples. The consumption of high δ¹⁵N-valued fish and birds can be responsible for this clustering.     

Spatial and Temporal Variations in Stable Carbon (δ13C) and Nitrogen (δ15N) Isotopic Composition of Symbiotic Scleractinian Corals

Tropical scleractinian corals are considered autotrophic as they rely mainly on photosynthesis-derived nutrients transferred from their photosymbionts. Corals are also able to capture and ingest suspended particulate organic matter, so heterotrophy can be an important supplementary trophic pathway to optimize coral fitness. The aim of this in situ study was to elucidate the trophic status of 10 coral species under contrasted environmental conditions in a French Polynesian lagoon. Carbon (δ¹³C) and nitrogen (δ¹⁵N) isotopic compositions of coral host tissues and photosymbionts were determined at 3 different fringing reefs during wet and dry seasons. Our results highlighted spatial variability in stable isotopic compositions of both coral host tissues and photosymbionts. Samples from the site with higher level of suspended particulate matter were ¹³C-depleted and ¹⁵N-enriched relative to corals and photosymbionts from less turbid sites. However, differences in both δ¹³C and δ¹⁵N between coral host tissues and their photosymbionts (Δ^{host-photosymbionts 13}C and Δ^{host-photosymbionts 15}N) were small (0.27 ± 0.76‰ and 1.40 ± 0.90‰, respectively) and similar at all sites, thus indicating no general increases in the heterotrophic pathway. Depleted δ¹³C and enriched δ¹⁵N values of coral host tissues measured at the most turbid site were explained by changes in isotopic composition of the inorganic nutrients taken up by photosymbionts and also by changes in rate of isotopic fractionation with environmental conditions. Our results also highlighted a lack of significant temporal variations in δ¹³C and δ¹⁵N values of coral host and photosymbiont tissues and in Δ^{host-photosymbionts 13}C and Δ^{host-photosymbionts 15}N values. This temporal stability indicated that corals remained principally autotrophic even during the wet season when photosymbiont densities were lower and the concentrations of phytoplankton were higher. Increased coral heterotrophy with higher food availability thus appears to be species-specific.     

Identifying carbon sources and trophic position of coral reef fishes using diet and stable isotope (δ15N and δ13C) analyses in two contrasted bays in Moorea, French Polynesia

Stable isotope ratios (δ¹⁵N and δ¹³C) and diet of three fish species, Stegastes nigricans, Chaetodon citrinellus and Epinephelus merra, were analyzed on the fringing coral reefs of two bays that are differentially exposed to river runoff on Moorea Island, French Polynesia. S. nigricans and C. citrinellus relied mostly on turf algae and presented similar trophic levels and δ¹⁵N values, whereas E. merra fed on large invertebrates (crabs and shrimps) and had higher trophic levels and δ¹⁵N values. Discrepancies existed between stomach content and stable isotope analyses for the relative importance of food items. Bayesian mixing models indicated that sedimented organic matter was also an important additional food for S. nigricans and C. citrinellus, and fishes for E. merra. The main sources of organic matter involved in the food webs ending with these species were algal turfs and surface sediments, while water particulate organic matter was barely used. Significant spatial differences in C and N isotopic ratios for sources and fishes were found within and between bays. Lower ¹³C and higher ¹⁵N values were observed for various compartments of the studied trophic network at the end of each bay than at the entrance. Differences were observed between bays, with organic sources and consumers being, on average, slightly more ¹³C-depleted and ¹⁵N-enriched in Cook’s Bay than in Opunohu Bay, linked with a higher mean annual flow of the river at Cook’s Bay. Our results suggest that rivers bring continental material into these two bays, which is partly incorporated into the food webs of fringing coral reefs at least close to river mouths. Thus, continental inputs can influence the transfer of organic matter within coral reef food webs depending on the diet of organisms.     

Elemental (C,N, P) and isotopic (δ13C, δ15N) signature of primary producers and their contribution to the organic matter in coastal lagoon sediment

Estuarine ecosystems are easily deteriorated by organic pollution because of its high primary productivity. To identify chemical proxies for the possible sources of autochthonous organic matter [phytoplankton-derived particulate organic material (POM), macroalgae and seagrass], we measured C:N:P and the ratios of carbon and nitrogen stable isotopes (δ¹³C and δ¹⁵N values) in two estuarine environments, the polyhaline lagoon, Lake Nakaumi, and the oligohaline lagoon, Lake Shinji, in Japan. Due to vigorous photosynthesis, the δ¹³C of phytoplankton-derived POM in Lake Nakaumi was larger than what would normally be expected from estuarine salinity gradients. Concentrations of nitrogen and phosphorus did not affect the δ¹³C of phytoplankton-derived POM. The δ¹⁵N of all plants was uniform and was higher than the δ¹⁵N of sediments. The seagrass showed a higher C:N ratio than POM and macroalgae, while the macroalgae showed a higher N:P ratio. Thus, simultaneous evaluation of C:N and N:P ratios would distinguish these three plant groups, and it would be possible to identify the source plants from the elemental ratios of the sediments.     

Patterns of δ~(13) C and δ~(15) N in wolverine Gulo gulo tissues from the Brooks Range,Alaska

Knowledge of carnivore diets is essential to understand how carnivore populations respond demographically to variations in prey abundance.Analysis of stable isotopes is a useful complement to traditional methods of analyzing carnivore diets.We used data on δ 13 C and δ 15 N in wolverine tissues to investigate patterns of seasonal and annual diet variation in a wolverine Gulo gulo population in the western Brooks Range,Alaska,USA.The stable isotope ratios in wolverine tissues generally reflected that of terr...     

Functional diversity in an Amazonian rainforest of French Guyana: a dual isotope approach (δ15N and δ13C)

Functional aspects of biodiversity were investigated in a lowland tropical rainforest in French Guyana (5°2′N, annual precipitation 2200 mm). We assessed leaf δ¹⁵N as a presumptive indicator of symbiotic N₂ fixation, and leaf and wood cellulose δ¹³C as an indicator of leaf intrinsic water-use efficiency (CO₂ assimilation rate/leaf conductance for water vapour) in dominant trees of 21 species selected for their representativeness in the forest cover, their ecological strategy (pioneers or late successional stage species, shade tolerance) or their potential ability for N₂ fixation. Similar measurements were made in trees of native species growing in a nearby plantation after severe perturbation (clear cutting, mechanical soil disturbance). Bulk soil δ¹⁵N was spatially quite uniform in the forest (range 3–5‰), whereas average leaf δ¹⁵N ranged from −0.3‰ to 3.5‰ in the different species. Three species only, Diplotropis purpurea, Recordoxylon speciosum (Fabaceae), and Sclerolobium melinonii (Caesalpiniaceae), had root bacterial nodules, which was also associated with leaf N concentrations higher than 20 mg g⁻¹. Although nodulated trees displayed significantly lower leaf δ¹⁵N values than non-nodulated trees, leaf δ¹⁵N did not prove a straightforward indicator of symbiotic fixation, since there was a clear overlap of δ¹⁵N values for nodulated and non-nodulated species at the lower end of the δ¹⁵N range. Perturbation did not markedly affect the difference δ¹⁵N_soil−δ¹⁵N_leaf, and thus the isotopic data provide no evidence of an alteration in the different N acquisition patterns. Extremely large interspecific differences in sunlit leaf δ¹³C were observed in the forest (average values from −31.4 to −26.7‰), corresponding to intrinsic water-use efficiencies (ratio CO₂ assimilation rate/leaf conductance for water vapour) varying over a threefold range. Wood cellulose δ¹³C was positively related to total leaf δ¹³C, the former values being 2–3‰ higher than the latter ones. Leaf δ¹³C was not related to leaf δ¹⁵N at either intraspecific or interspecific levels. δ¹³C of sunlit leaves was highest in shade hemitolerant emergent species and was lower in heliophilic, but also in shade-tolerant species. For a given species, leaf δ¹³C did not differ between the pristine forest and the disturbed plantation conditions. Our results are not in accord with the concept of existence of functional types of species characterized by common suites of traits underlying niche differentiation; rather, they support the hypothesis that each trait leads to a separate grouping of species. Received: 18 August 1997 / Accepted: 14 April 1998     

Effects of anticoagulants on stable-isotope values (δ13C and δ15N) of shark blood components

The effects of anticoagulant EDTA and sodium heparin (SH) on stable carbon δ¹³C and nitrogen δ¹⁵N isotopic values of red blood cells (RBC) and blood plasma in juvenile blacktip reef sharks Carcharhinus melanopterus were analysed. Plasma preserved with anticoagulants was not isotopically distinct from plasma stored in no-additive control tubes but RBC δ¹⁵N values exhibited small enrichments when preserved with EDTA and SH. Results suggest EDTA and SH are viable anticoagulants for stable isotopic analyses of blood fractions but further studies are advised to validate results.     

Natal origins and timing of migration of two passerine species through the southern Alps: inferences from multiple stable isotopes (δ2H, δ13C, δ15N, δ34S) and ringing data

Understanding spatial linkages between areas used by migratory animals during the annual cycle is fundamental to their conservation. Stable isotope measurements of animal tissues can be a valuable tool in understanding spatial connectivity and migration phenology of migratory wildlife. We inferred natal origins of two migratory passerines, European Pied Flycatcher Ficedula hypoleuca and European Robin Erithacus rubecula, captured during autumn migration in the Italian Alps, by combining feather δ²H (δ²H_f) and ring recovery data. We used a spatially explicit likelihood-based method to assign individuals to a precipitation δ²H surface calibrated to represent feather δ²H, together with the directional probability of origin from ring recoveries. The highest probabilities of origin for most individuals of both species were in central and north-eastern Europe. Seasonal trends in δ²H_f, which described the species’ migratory phenology through the Italian Alps, were correlated with feather δ¹³C, δ¹⁵N and δ³⁴S values, indicating strong spatial discrimination related to continental patterns for these isotopes. We demonstrate how this combined information can define catchment areas and migratory connectivity of birds intercepted in the Alps. We highlight the importance of ringing data in defining directional priors to define Bayesian-based probability surfaces using continental δ²H_f isoscapes, and how such information can be used to inform estimates of migratory connectivity.