Use Of Stable Isotope Analysis Research Articles

Diet quality and muscle tissue location influence consumer-diet discrimination in captive-reared rock lobsters (Panulirus cygnus)

Fundamental to the accuracy of stable isotope analysis in trophodynamic studies is the ability to predict discrimination between a consumer and its diet. Despite the widespread use of stable isotope analysis in trophic ecology, uncertainty still surrounds the factors affecting consumer-diet discrimination. Here we present evidence that diet quality and location of muscle tissue analysed affects the consumer-diet discrimination for the western rock lobster, Panulirus cygnus. Consumer-diet δ15N and δ13C discrimination for western rock lobster tail tissue were 1.67–2.97 and 2.92–3.60‰, respectively, with δ13C discrimination differing to values reported in the literature. Differences in nitrogen and carbon discrimination were observed between tail and leg tissue of lobsters of 1.22 and 1.13‰, respectively. Diet quality was also found to affect consumer-diet discrimination, with high protein pilchard diet leading to lower δ15N and higher δ13C discrimination. Diet quality should be considered as a factor that has the potential to affect consumer-diet discrimination when interpreting results from stable isotope studies.

Can stable isotope (δ13C and δ15N) measurements of little auk (Alle alle) adults and chicks be used to track changes in high-Arctic marine foodwebs?

The little auk (Alle alle), a small and abundant planktivorous seabird that breeds in the high Arctic, has the potential to be used as a monitor of the composition and abundance of lower trophic-level zooplankton. We investigated age- and sex-related sources of variation in diet and stable isotope (δ13C and δ15N) values of little auks breeding in Spitsbergen during the summer of 2002 to evaluate this possibility. Stable isotope profiles of both adult and chick blood changed over the breeding season, with blood δ15N values increasing and δ13C values decreasing. This could represent a switch to higher trophic-level prey derived from more pelagic sources. However, while chick blood δ13C values followed those values in their meals, this was not the case for blood δ15N values, suggesting additional physiological mechanisms influencing blood δ15N values in growing chicks. Chicks had consistently lower δ15N values than their parents, which may indicate they were being fed on lower trophic-level prey items or may alternatively reflect complexities in chick blood δ15N values through the growth period. These results have several important implications for use of stable isotope analysis as a tool to detect changes in seabird diet and availability of lower trophic-level prey in high-Arctic marine environments. Until physiological aspects of stable isotope discrimination are well understood, we caution against using chicks of this seabird as any form of isotopic monitor.

Use of stable isotope analysis to evaluate the possible impact of fish migration on a lake biomanipulation

1. A lake restoration project involving mass fish removals was started to improve further the water quality of a lake recovering from severe eutrophication. The effectiveness of such biomanipulation as a lake management option could be compromised if removed fish are rapidly replaced by large-scale immigration from a connected lake. 2. Stable isotope analysis (SIA) was used to examine the migrations of perch (Perca fluviatilis L.) and roach (Rutilus rutilus (L.)) between two connected lakes in central Finland, Jyvasjarvi and the larger Paijanne. Baseline δ15N signatures in primary consumers were significantly higher in Jyvasjarvi and this difference was reflected further up the food chain in fish. 3. Spring fish spawning migrations from Paijanne resulted in a significant decrease in mean δ15N signatures of both perch and roach in the adjacent part of Jyvasjarvi, but by summer these mean δ15N signatures in fish muscle tissue had increased again to match those in the rest of the lake. δ13C signatures of both fish species were more variable and were of less value for tracking fish migrations. 4. A robust discriminant analysis was used to classify perch and roach caught from Jyvasjarvi into fish originating from either Jyvasjarvi or Paijanne. The greatest proportions of immigrant perch and roach in Jyvasjarvi were observed in spring 2004 (71% and 56%), when biomanipulation of Jyvasjarvi had started. Immigrant fish percentages in the following spring were considerably lower (24% and 14%, respectively), most likely dueto effective biomanipulation. 5. Results indicate that the current biomanipulation has decreased the extent of the spring migration, and that immigrant fish have not significantly repopulated Jyvasjarvi after the intense fishing. More generally, the data illustrate how stable isotopes offer the possibility to track migrations of common fish species between adjacent freshwater lakes. Copyright © 2007 John Wiley & Sons, Ltd.

Influence of morphology and amphibious life‐style on the feeding ecology of the mudskipper Periophthalmus argentilineatus

The potential feeding advantages that are offered to the barred mudskipper Periophthalmus argentilineatus by its amphibious life‐style were investigated. To this end the feeding ecology of these fish, the position they occupy in the mangrove food web and the importance of different morphological factors for their diet were studied. Analysis of gut contents showed that the major component of the barred mudskipper diet changes during growth from small crustaceans (e.g. copepods and amphipods) to polychaetes to mangrove crabs. This dietary shift was found to be facilitated through enlargement of the mouth, as well as elongation of the gut. The use of stable isotope analysis revealed that barred mudskippers had a similar trophic position as other zoobenthivore fish species, but that their diet did not appear to be affected by competition for food with other zoobenthivores. The amphibious habits of the barred mudskipper provide specific advantages to its feeding ecology, such as access to an abundant food source of mangrove crabs, which are mostly inaccessible for other fish species.

Evaluating bone collagen extraction methods for stable isotope analysis in dietary studies

The specific purpose of this study was to compare three different collagen extraction methods commonly used in isotope laboratories conducting dietary studies. We evaluated their resultant differences in δ 13C and δ 15N, collagen quality and collagen yield. Our study was based on well-preserved skeletal material from the medieval period in Denmark. Our study shows that there is a systematic significant difference in the yield and the δ 13C values between the three methods. Using the method of DeNiro and Epstein [DeNiro, M.J., Epstein, S., 1981. Influence of diet on the distribution of nitrogen isotopes in animals. Geochimica et Cosmochimica Acta 45, 341–351] with NaOH as cleaning agent, will, according to our study, give δ 13C values that are on average ±0.32‰ more positive than using the ultra-filtration method [Brown, T.A., Nelson, D.E., Vogel, J.S., Southon, J.R., 1988. Improved collagen extraction by modified Longin method. Radiocarbon 30 (2), 171–177, modified in Richards, M.P., Hedges R.E.M., 1999. Stable isotope evidence for similarities in the types of marine foods used by late Mesolithic humans at sites along the Atlantic coast of Europe. Journal of Archaeological Science 26, 717–722]. The third method, which is a modified version of the second method, excluded the ultra-filtration step. This method seems to give δ 13C values that lie in between the other methods. Our study did not show any significant difference in δ 15N values. Although the differences between the methods are very small, we conclude that the use of stable isotope analysis in food determination studies requires adherence to routine methods for preparing and measuring samples.

Niche variation in burying beetles (Nicrophorusspp.) associated with marine and terrestrial carrion

Ecological opportunity can influence niche variation within and among species. Forensic reconstruction of diet has been made possible with the now widespread use of stable isotope analysis, although it has not to date been applied to communities based on carrion resources. Within a salmon-bearing watershed in coastal British Columbia, we reconstructed the dietary niches of two burying beetle species (Silphidae: Nicrophorus investigator Zetterstedt, 1824 and Nicrophorus defodiens Mannerheim, 1846) using stable isotopes (δ15N, δ13C). We separated available carrion sources for beetles of chum salmon ( Oncorhynchus keta (Walbaum, 1792)) and pink salmon ( Oncorhynchus gorbuscha (Walbaum, 1792)), the shrew Sorex monticolus Merriam, 1890, songbirds ( Troglodytes troglodytes (L., 1758), Catharus ustulatus (Nuttall, 1840), Catharus guttatus (Pallas, 1811)), and black-tailed deer ( Odocoileus hemionus hemionus (Rafinesque, 1817)) into three isotopically homogeneous subsets: (1) salmon, (2) shrews and songbirds, and (3) deer. The majority (86.5%) of N. investigator individuals were raised on a diet of salmon carrion, while 100% of N. defodiens individuals had a larval diet consistent with carrion from shrews and songbirds. Larger isotopic variance predicts wider dietary niches, which may be useful for testing functional ecological variation within and among species.

Applications, Considerations, and Sources of Uncertainty When Using Stable Isotope Analysis in Ecotoxicology

Stable isotope analysis (SIA) has become a powerful tool for ecotoxicologists to study dietary exposure and biomagnification of contaminants in wild animal populations. The use of SIA in ecotoxicology continues to expand and, while much more is known about the mechanisms driving patterns of isotopic ratios in consumers, there remain several considerations or sources of uncertainty that can influence interpretation of data from field studies. We outline current uses of SIA in ecotoxicology, including estimating the importance of dietary sources of carbon and their application in biomagnification studies, and we present six main considerations or sources of uncertainty associated with the approach: (1) unequal diet-tissue stable isotope fractionation among species, (2) variable diet-tissue stable isotope fractionation within a given species, (3) different stable isotope ratios in different tissues of the animal, (4) fluctuating baseline stable isotope ratios across systems, (5) the presence of true omnivores, and (6) movement of animals and nutrients between food webs. Since these considerations or sources of uncertainty are difficult to assess in field studies, we advocate that researchers consider the following in designing ecotoxicological research and interpreting results: assess and utilize variation in stable isotope diet-tissue fractionation among animal groups available in the literature; determine stable isotope ratios in multiple tissues to provide a temporal assessment of feeding; adequately characterize baseline isotope ratios; utilize stomach contents when possible; and assess and integrate life history of study animals in a system.

The effect of growth rate on tissue‐diet isotopic spacing in rapidly growing animals. An experimental study with Atlantic salmon (Salmo salar)

The difference in isotopic composition between a consumer's tissues and that of its diet is a critical aspect of the use of stable isotope analyses in ecological and palaeoecological studies. In a controlled feeding experiment with the Atlantic salmon, Salmo salar, we demonstrate for the first time that the value of tissue-diet isotope spacing in nitrogen in a growing animal is not constant, but varies inversely with growth rate. The value of tissue-diet isotopic spacing in N reflects N use efficiency. Thus, in salmon, growth rate is accompanied by, or requires, increased N use efficiency. The total range in tissue-diet isotopic spacing in N seen in the experimental population of 25 fish was 1 per thousand, approximately 50% of the total trophic shift. Mean equilibrium tissue-diet isotopic spacing (+/-standard deviation) in salmon averaged 2.3 per thousand (+/-0.3 per thousand) and 0.0 per thousand (+/-0.3 per thousand) for N in muscle and liver, respectively, and 2.1 per thousand (+/-0.1 per thousand) and 1.6 per thousand (+/-0.3 per thousand) for C in muscle and liver, respectively. Feeding with a mixed dietary source (wheat and fish-meal origin) resulted in tissue-diet isotopic fractionation in both C and N due to the differential digestibility of food components with distinct isotopic composition. The rate of change in isotopic composition of S. salar tissues was dominated by growth, but the estimated contribution of metabolic turnover to change in tissue N was relatively high for an ectothermic animal at ca. 20-40%. The estimated half-life for metabolic turnover of the tissue N pool was ca. 4 months in both muscle and liver tissue. This is the first study to demonstrate a direct relationship between tissue-diet isotopic spacing in N and growth rate and adds to the growing list of factors known to influence the level of isotopic separation between a consumer's tissue and that of its diet.

Controlling for anthropogenically induced atmospheric variation in stable carbon isotope studies

Increased use of stable isotope analysis to examine food-web dynamics, migration, transfer of nutrients, and behavior will likely result in expansion of stable isotope studies investigating human-induced global changes. Recent elevation of atmospheric CO2 concentration, related primarily to fossil fuel combustion, has reduced atmospheric CO2 delta13C (13C/12C), and this change in isotopic baseline has, in turn, reduced plant and animal tissue delta13C of terrestrial and aquatic organisms. Such depletion in CO2 delta13C and its effects on tissue delta13C may introduce bias into delta13C investigations, and if this variation is not controlled, may confound interpretation of results obtained from tissue samples collected over a temporal span. To control for this source of variation, we used a high-precision record of atmospheric CO2 delta13C from ice cores and direct atmospheric measurements to model modern change in CO2 delta13C. From this model, we estimated a correction factor that controls for atmospheric change; this correction reduces bias associated with changes in atmospheric isotopic baseline and facilitates comparison of tissue delta13C collected over multiple years. To exemplify the importance of accounting for atmospheric CO2 delta13C depletion, we applied the correction to a dataset of collagen delta13C obtained from mountain lion (Puma concolor) bone samples collected in California between 1893 and 1995. Before correction, in three of four ecoregions collagen delta13C decreased significantly concurrent with depletion of atmospheric CO2 delta13C (n > or = 32, P < or = 0.01). Application of the correction to collagen delta13C data removed trends from regions demonstrating significant declines, and measurement error associated with the correction did not add substantial variation to adjusted estimates. Controlling for long-term atmospheric variation and correcting tissue samples for changes in isotopic baseline facilitate analysis of samples that span a large temporal range.

Stable isotopes in ecological studies

Stable isotope ratio analyses at natural abundance levels are an increasingly useful tool in ecological studies, with new papers being published in nearly every issue of Oecologia and other ecological journals. In April 2004, the National Institute of Water and Atmospheric Research (NIWA) and the Institute of Geological and Nuclear Sciences (IGNS) co-hosted the fourth conference on ‘‘Applications of Stable Isotopes to Ecological Studies’’ at the Museum of New Zealand Te Papa Tongarewa, Wellington, New Zealand (http:// www.isoecol.org/previous.html). This biannual international conference brings together a wide range of biologists, physiologists, geochemists, and environmental scientists who use stable isotopes to identify and solve gaps in ecological knowledge. Topics presented at this meeting were very diverse, and included the use of stable isotope analyses in ecological management of terrestrial, marine and freshwater systems, the hydrologic cycle, the carbon cycle and primary productivity, microbial ecology, nutrient cycling, pollution ecology, precipitation analyses across broad spatial scales, terrestrial and aquatic trophic level and diet studies, bird migration studies, and palaeoecology. The need for these international meetings continues to grow since the first meeting held in 1998 in Saskatoon, Canada (Hobson and Wassenaar 1999); the next conference is planned for Belfast, Northern Ireland in August 2006 (http://www.isoecol.org). A number of contributors to the New Zealand conference jointly submitted manuscripts for review and publication in Oecologia. Having now satisfactorily completed the review process, a number of the conference contributions are published together in this issue of Oecologia. These papers highlight the diverse ecological studies discussed in New Zealand. Bird migration studies take advantage of the pronounced isotope gradients across continents (Bowen et al. 2005; Hobson et al. 2004; Pain et al. 2004). Mazerolle et al. (2005) developed the use of stable hydrogen isotopes of feather and claw material from white-throated sparrows (Zonotrichia albicollis) in order to elucidate both breeding and wintering catchment areas, an approach augmented by conventional bird-banding data. Wunder et al. (2005) provided a quantitative assessment on the utility of using isotope tracers in feathers of known origin. There is a long history of using stable isotopes in marine studies. Continuing this area of investigation, Hicks et al. (2005) investigated the contribution of marine-derived nutrients from spawning coho salmon (Oncorhynchus kisutch) to freshwater-riparian foodwebs in southcentral Alaska, and found that spawning salmon provide marine nitrogen and carbon comprising about 35% of the dietary needs of juvenile salmon. Harrod et al. (2005) examined stable isotope analyses in the evaluation of ecological plasticity in eels. Cunjak et al. (2005) showed the utility of using isotopes as trackers of fish movements, especially when combined with more traditional approaches. Isotopes are often useful tracers in ecological systems. Connolly et al. (2005) examined relationships in carbon dynamics by invertebrates in estuaries. Wissell and Fry Communicated by Jim Ehleringer

Natural attenuation and enhanced bioremediation of organic contaminants in groundwater

An area of intense scientific and practical interest is the biogeochemical and microbial processes determining the success of natural attenuation, biostimulation and/or bioaugmentation treatments for organic contaminants in groundwater. Recent studies in this area have focused on the reductive dechlorination of chlorinated solvents, the degradation of the fuel additive methyl tert-butyl ether, and the removal of long-term hydrocarbon contamination. These studies have been facilitated by the use of stable isotope analysis to demonstrate in situ bioremediation and push-pull tests, in which isotopes are injected into aquifers and then quickly retrieved and analyzed, to measure in situ activity. Molecular tools such as quantitative PCR, the detection of mRNA expression, and numerous DNA fingerprinting methods have also proved valuable, being employed to identify and sometimes quantify environmentally important organisms or changes in communities. Methods to track bacteria and tools to characterize bacterial attachment properties have also offered insight into bacterial transport in situ.

Isotope variations in white-tailed kites from various habitats in California: Possible limitations in assessing prey utilization and population dynamics

White-tailed kite (Elanus leucurus) populations in the 1930s were close to extirpation in the United States. But by the 1940s, an upward trend towards recovery was apparent and continued to their current stable population levels. These dramatic fluctuations in kite numbers may have been related to changes in rodent prey populations due to the conversion of native habitats to agriculture. To address this question, we evaluated the use of stable isotope analysis in determining if a shift in diet could be isotopically differentiated in current and historic kite populations. We first compared δ 13C, δ 15N, and δ 34S values from present-day kite flight feathers and prey fur samples from four locations in California. The total ranges of isotope values for kite and their rodent prey were similar within each site. Carbon isotope values ranged from m 27.1 to m 22.2 in Arcata, m 26.1 to m 16.9 in Davis, m 27.0 to m 15.0 in Cosumnes, and m 28.2 to m 11.6 in Santa Barbara. Nitrogen isotope values ranged from 3.2 to 15.7 in Arcata, 2.8 to 12.7 in Davis, 4.0 to 15.7 in Cosumnes, and 1.7 to 20.0 in Santa Barbara. Sulfur isotope values ranged from m 7.8 to 12.4 in Arcata, m 1.1 to 9.2 in Davis, 0.7 to 10.9 in Cosumnes, and m 8.6 to 15.6 in Santa Barbara. Carbon, nitrogen, and sulfur isotope values at each site reflect typical trophic enrichments due to physiological processes. At each site, δ 13C and δ 15N values reflected the influence of a predominantly C3 or a mixed C3/C4 plant community. Sulfur isotope values reflect the influence of predominant marine or terrestrial sulfur sources at each site. However, variability in isotope values may limit the usefulness of such analyses for addressing prey utilization and population dynamics.

Using stable isotopes to assess seasonal patterns of avian predation across a terrestrial-marine landscape.

In this study, we used the stable isotope ratios 15N/14N and 13C/12C to clarify the spatial and temporal patterns of small mammal derpredation by wide-ranging raptors across a salt marsh and grassland landscape. To determine whether clear isotope signals existed for the two adjoining habitats, and if these differences could allow for an assessment of the seasonal habitat origins of depredated rodents, primarily the vole, Microtus californicus, we investigated the nitrogen and carbon isotope ratios for three sets of data. First, to establish potential differences in isotope signatures between the two habitats, we collected plant species consumed by the vole. Second, we analyzed bone collagen from voles originating from known locations along a gradient from marsh to grassland. Finally, the signatures of these unpredated voles were then used to categorize isotope values of vole remains found in the pellets of their raptor predators. Results indicated that mean δ15N and δ13C were both higher in marsh plants than in grassland species. Although both isotopes showed a trophic level increase from plant to bone collagen, nitrogen provided the more informative signature when attempting to distinguish the habitat origins of voles. We found that the δ15N values of unpredated voles were significantly related to the distance of the specimen along the marsh-grassland interface, with higher values in the marsh habitat. Analysis of the depredated bones found in raptor pellets indicated that a greater percentage of voles originated in grassland during the spring/summer season (88%), as compared with the fall/winter period (60%). Finally, we compared this pattern of habitat and season-specific predation to another measure of predation risk, the relative encounter rates of raptors foraging for voles, and found that the two estimates were similar except during the winter season. This study demonstrates the usefulness of stable isotope analysis for elucidating seasonal patterns of predation across a complex landscape.

Rates and Mechanisms of Isotopic Exchange

### Background The proliferation of stable isotope laboratories in recent years has led to a marked increase in the routine use of stable isotope analyses in geochemical studies. This trend is likely to continue because of the development of micro-analytical techniques (e.g. laser ablation, ion microprobe, femtomole-carrier gas methods, etc.) for stable isotope analysis and the potential of these techniques to reveal the detailed sequence of thermal and fluid histories preserved in the rock record (see McKibben et al. 1998). Equilibrium isotope fractionation factors and rates of isotopic exchange form the cornerstones for the interpretation of stable isotope data from natural systems. Indeed, determination of the distribution of the stable isotopes of C-O-H-S in gases, fluids, and minerals has become a standard and extremely powerful approach in elucidating the temperatures, material fluxes, rock and fluid origins, and time scales associated with ancient and active fluid-rock interaction processes in the Earth’s crust (see Valley et al. 1986). Although the thermodynamic principles underlying isotopic fractionation behavior were developed more than fifty years ago by Urey (1947), the calibration of fractionation factors has proven to be difficult. During the last four decades, a great deal of progress has been made in determining the equilibrium fractionations of 18O/16O, D/H, 13C/12C, and 34S/32S among fluid and mineral components at high temperatures (≥400°C), both from experimental exchange studies, theoretical calculations (using vibrational spectra), semi-empirical calculations (using bond-type considerations), and empirical calibrations based on natural assemblages (see references in O’Neil 1986; Kyser 1987; Criss 1991, 1999; Chacko et al., this volume). Of these, the experimental method is the most direct because it involves the least number of a priori assumptions. Despite the utility of the equilibrium approach in quantifying the isotopic behavior in select water-rock …

Tracing Nutrient Allocation to Reproduction With Stable Isotopes: A Preliminary Investigation Using Colonial Waterbirds of Great Slave Lake

AbstractWe investigated the use of stable-isotope analysis as a direct means of tracing allocation of endogenous protein and lipid reserves to reproduction in five gulls (Larus canus, L. delawarensis, L. californicus, L. argentatus, L. philadelphia), four terns (Sterna caspia, S. hirundo, S. paradisaea, Chlidonias niger), and one jaeger (Stercorarius parasiticus) breeding on Great Slave Lake (GSL) in the Northwest Territories. Our approach was based on assumptions that (1) body tissues of birds just arriving at GSL from their assumed marine-associated wintering habitats would have stable-isotope ratios more enriched than those of birds in equilibrium with the local GSL foodweb, and (2) mobilization of these reserves to reproduction could be traced by the isotopic measurement of egg macronutrients. As predicted, the pectoral muscle of six species of arriving birds was more enriched in 13C (x̄ = −21.5‰) and 15N (x̄ = 12.7‰) than was that of postbreeding birds (13C, x̄ = −23.5‰; 15N, x̄ = 9.9‰) or hatching-year birds raised at GSL (13C, x̄ = −24.3‰; 15N, x̄ = 9.0‰). Abdominal fat of arriving Herring Gulls and Mew Gulls was more enriched in 13C (x̄ = −19.7‰) than the fat of other species (x̄ = −23.4‰), indicating lipids of marine origin. We compared isotope values of the local GSL foodweb with dietary values predicted from isotope measurements of egg macronutrients if diets were entirely derived at GSL. Isotopic analysis of lipid-free egg yolk, yolk lipid, and shell carbonate suggested that for most species, little if any endogenous protein reserves were used for reproduction, with the possible exception of Caspian Terns, whose egg protein and egg lipid values, and Common Terns, whose egg protein values, were more enriched in 13C than those of the other species. Although endogenous nutrient reserves likely were important to birds during migration and the initial settling period at GSL, local food supplies were adequate to provide nutrients for reproduction.

TRACING NUTRIENT ALLOCATION TO REPRODUCTION WITH STABLE ISOTOPES: A PRELIMINARY INVESTIGATION USING COLONIAL WATERBIRDS OF GREAT SLAVE LAKE

We investigated the use of stable-isotope analysis as a direct means of tracing allocation of endogenous protein and lipid reserves to reproduction in five gulls (Larus canus, L. delawarensis, L. californicus, L. argentatus, L. philadelphia), four terns (Sterna caspia, S. hirundo, S. paradisaea, Chlidonias niger), and one jaeger (Stercorarius parasiticus) breeding on Great Slave Lake (GSL) in the Northwest Territories. Our approach was based on assumptions that (1) body tissues of birds just arriving at GSL from their assumed marine-associated wintering habitats would have stable-isotope ratios more enriched than those of birds in equilibrium with the local GSL foodweb, and (2) mobilization of these reserves to reproduction could be traced by the isotopic measurement of egg macronutrients. As predicted, the pectoral muscle of six species of arriving birds was more enriched in 13C (x̄ = −21.5‰) and 15N (x̄ = 12.7‰) than was that of postbreeding birds (13C, x̄ = −23.5‰; 15N, x̄ = 9.9‰) or hatching-year birds raised at GSL (13C, x̄ = −24.3‰; 15N, x̄ = 9.0‰). Abdominal fat of arriving Herring Gulls and Mew Gulls was more enriched in 13C (x̄ = −19.7‰) than the fat of other species (x̄ = −23.4‰), indicating lipids of marine origin. We compared isotope values of the local GSL foodweb with dietary values predicted from isotope measurements of egg macronutrients if diets were entirely derived at GSL. Isotopic analysis of lipid-free egg yolk, yolk lipid, and shell carbonate suggested that for most species, little if any endogenous protein reserves were used for reproduction, with the possible exception of Caspian Terns, whose egg protein and egg lipid values, and Common Terns, whose egg protein values, were more enriched in 13C than those of the other species. Although endogenous nutrient reserves likely were important to birds during migration and the initial settling period at GSL, local food supplies were adequate to provide nutrients for reproduction.

Tracing origins and migration of wildlife using stable isotopes: a review.

To understand the ecology of migratory animals it is important to link geographic regions used by individuals including breeding, wintering, and intermediate stopover sites. Previous conventional approaches used to track animal movements have relied on extrinsic markers and typically the subsequent recovery of individuals. This approach has generally been inappropriate for most small, or non-game animals. The use of intrinsic markers such as fatty acid profiles, molecular DNA analyses, and the measurement of naturally occurring stable isotopes in animal tissues offer alternative approaches. This paper reviews the use of stable isotope analyses (primarily δ13C, δ15N, δ34S, δD, δ87Sr) to trace nutritional origin and migration in animals. This approach relies on the fact that foodweb isotopic signatures are reflected in the tissues of organisms and that such signatures can vary spatially based on a variety of biogeochemical processes. Organisms moving between isotopically distinct foodwebs can carry with them information on the location of previous feeding. Such an approach has been used to track animal use of inshore versus offshore, marine versus freshwater, terrestrial C3 versus marine, terrestrial mesic versus xeric, and C3 versus C4 or Crassulacean acid metabolism foodwebs. More recently, the use of stable hydrogen isotope analyses (δD) to link organisms to broad geographic origin in North America is based on large-scale isotopic contours of growing-season average δD values in precipitation. This technique, especially when combined with the assay of other stable isotopes, will be extremely useful in helping to track migration and movement of a wide range of animals from insects to birds and mammals. Future research to refine our understanding of natural and anthropogenic-induced isotopic gradients in nature, and to explore the use of stable isotopes of other elements, is recommended.

Analysis of Stable Isotope Data: A K Nearest-Neighbors Randomization Test

The use of stable isotope analysis in ecological and wildlife studies is rapidly increasing. Studies include evaluating flow of nutrients in ecosystems and studying dietary composition of individual animals. Several mixing models have been developed to evaluate the relative contribution of different foods to the diet of consumers. All these mixing models require that all prey types will be significantly different in bivariate space. This requirement usually poses a problem in analyzing data of stable isotope ratios because sample sizes in most studies are small and seldom normally distributed. We propose a randomization test that we based on the K nearest-neighbor approach. Results from our simulations of power revealed that the K nearest-neighbor test appears to have high power even with small sample sizes and comparatively low displacement. The K nearest-neighbor test described here provides the preliminary statistical analysis necessary for the use of the mixing models, and therefore is a new, powerful tool for analyzing stable isotope data. In evaluating the test performance on data collected from American martens (Martes americana) and their prey on Chichagof Island, Southeast Alaska, we were able to reject our null hypothesis that all samples of prey were drawn from identical populations (P = 0.05). A program written in Pascal or S-Plus is available from the authors to evaluate the K nearest-neighbor statistic for several groups.

Dietary and environmental reconstruction with stable isotope analyses of herbivore tooth enamel from the Miocene locality of Fort Ternan, Kenya.

Tooth enamel of nine Middle Miocene mammalian herbivores from Fort Ternan, Kenya, was analyzed for delta 13C and delta 18O. The delta 18O values of the tooth enamel compared with pedogenic and diagenetic carbonate confirm the use of stable isotope analysis of fossil tooth enamel as a paleoenvironmental indicator. Furthermore, the delta 18O of tooth enamel indicates differences in water sources between some of the mammals. The delta 13C values of tooth enamel ranged from -8.6(-)-13.0/1000 which is compatible with a pure C3 diet, though the possibility of a small C4 fraction in the diet of a few of the specimens sampled is not precluded. The carbon isotopic data do not support environmental reconstructions of a Serengeti-typed wooded grassland with a significant proportion of C4 grasses. This study does not preclude the presence of C3 grasses at Fort Ternan; it is possible that C3 grasses could have had a wider geographic range if atmospheric CO2 levels were higher than the present values.

Use of stable isotopes to determine diets of living and extinct bears

The potential use of stable-isotope analyses (δ13C and δ15N) to estimate bear diets was assessed in 40-day feeding trials using American black bears (Ursus americanus). Bear plasma and red blood cells have half-lives of ~4 days and ~28 days, respectively. The isotopic signature of bear plasma is linearly related to that of the diet, and with the exception of adipose tissue, there is no isotopic fractionation across bear tissues. Isotopic analyses were used to estimate the diets of three bear populations: Pleistocene cave bears (U. speleaus) in Europe, grizzly bears (Ursus arctos horribilis) inhabiting the Columbia River drainage prior to 1931, and brown bears (U. arctos) of Chichagof and Admiralty islands, Alaska. Cave bears were omnivores with terrestrially produced meat contributing from 41 to 78% (58 ± 14%) of their metabolized carbon and nitrogen. Salmon contributed from 33 to 90% (58 ± 23%) of the metabolized carbon and nitrogen in grizzly bears from the Columbia River drainage. Finally, most brown bears on Chichagof and Admiralty islands feed upon salmon during the late summer and fall; however, a subpopulation of bears exists that does not utilize salmon.