The influence of diet on the distribution of nitrogen isotopes in animals was investigated by analyzing animals grown in the laboratory on diets of constant nitrogen isotopic composition. 
The isotopic composition of the nitrogen in an animal reflects the nitrogen isotopic composition of its diet. The δ^(15)N values of the whole bodies of animals are usually more positive than those of their diets. Different individuals of a species raised on the same diet can have significantly different δ^(15)N values. The variability of the relationship between the δ^(15)N values of animals and their diets is greater for different species raised on the same diet than for the same species raised on different diets. Different tissues of mice are also enriched in ^(15)N relative to the diet, with the difference between the δ^(15)N values of a tissue and the diet depending on both the kind of tissue and the diet involved. The δ^(15)N values of collagen and chitin, biochemical components that are often preserved in fossil animal remains, are also related to the δ^(15)N value of the diet. 
The dependence of the δ^(15)N values of whole animals and their tissues and biochemical components on the δ^(15)N value of diet indicates that the isotopic composition of animal nitrogen can be used to obtain information about an animal's diet if its potential food sources had different δ^(15)N values. The nitrogen isotopic method of dietary analysis probably can be used to estimate the relative use of legumes vs non-legumes or of aquatic vs terrestrial organisms as food sources for extant and fossil animals. However, the method probably will not be applicable in those modern ecosystems in which the use of chemical fertilizers has influenced the distribution of nitrogen isotopes in food sources. 
The isotopic method of dietary analysis was used to reconstruct changes in the diet of the human population that occupied the Tehuacan Valley of Mexico over a 7000 yr span. Variations in the δ^(15)C and δ^(15)N values of bone collagen suggest that C_4 and/or CAM plants (presumably mostly corn) and legumes (presumably mostly beans) were introduced into the diet much earlier than suggested by conventional archaeological analysis.

Influence of Diet On the Distribtion of Nitrogen Isotopes in Animals

The use of stable isotopes to solve biogeochemical problems in ecosystem analysis is increasing rapidly because stable isotope data can contribute both source-sink (tracer) and process information: The elements C, N, S, H, and all have more than one isotope, and isotopic compositions of natural materials can be measured with great precision with a mass spectrometer. Isotopic compositions change in predictable ways as elements cycle through the biosphere. These changes have been exploited by geochemists to understand the global elemental cycles. Ecologists have not until quite recently employed these techniques. The reasons for this are, first, that most ecologists do not have the background in chemistry and geochemistry to be fully aware of the possibilities for exploiting the natural variations in stable isotopic compositions, and second, that stable isotope ratio measurements require equipment not normally available to ecologists. This is unfortunate because some of the more intractable problems in ecology can be profitably addressed using stable isotope measurements. Stable isotopes are ideally suited to increase our understanding of element cycles in ecosystems. This review is written for ecologists who would like to learn more about how stable isotope analyses have been and can be used in ecosystem studies. We begin with an explanation of isotope terminology and fractionation, then summarize isotopic distributions in the C, N, and S biogeochemical cycles, and conclude with five case studies that show how stable isotope measurements can provide crucial information for ecosystem analysis. We restrict this review to studies of natural variations in C, N, and S isotopic abundances, cxcluding from consideration ~5N enrichment studies and hydrogen and oxygen isotope studies. Our focus on C, N, and S derives in part from our

Stable isotopes in ecosystem studies

Stable isotopes are increasingly being used as tracers in environmental studies. One application is to use isotopic ratios to quantitatively determine the proportional contribution of several sources to a mixture, such as the proportion of various pollution sources in a waste stream. In general, the proportional contributions of n+1 different sources can be uniquely determined by the use of n different isotope system tracers (e.g., δ13C, δ15N, δ18O) with linear mixing models based on mass balance equations. Often, however, the number of potential sources exceeds n+1, which prevents finding a unique solution of source proportions. What can be done in these situations? While no definitive solution exists, we propose a method that is informative in determining bounds for the contributions of each source. In this method, all possible combinations of each source contribution (0–100%) are examined in small increments (e.g., 1%). Combinations that sum to the observed mixture isotopic signatures within a small tolerance (e.g., ±0.1‰) are considered to be feasible solutions, from which the frequency and range of potential source contributions can be determined. To avoid misrepresenting the results, users of this procedure should report the distribution of feasible solutions rather than focusing on a single value such as the mean. We applied this method to a variety of environmental studies in which stable isotope tracers were used to quantify the relative magnitude of multiple sources, including (1) plant water use, (2) geochemistry, (3) air pollution, and (4) dietary analysis. This method gives the range of isotopically determined source contributions; additional non-isotopic constraints specific to each study may be used to further restrict this range. The breadth of the isotopically determined ranges depends on the geometry of the mixing space and the similarity of source and mixture isotopic signatures. A sensitivity analysis indicated that the estimated ranges vary only modestly with different choices of source increment and mass balance tolerance parameter values. A computer program (IsoSource) to perform these calculations for user-specified data is available at http://www.epa.gov/wed/pages/models.htm
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Source partitioning using stable isotopes: coping with too many sources.

Use of stable isotope techniques to quantify food web relationships requires a priori estimates of the enrichment or depletion in δ15N and δ13C values between prey and predator (known as trophic fractionation; hereafter Δδ15N and Δδ13C). We conducted a broad-scale analysis of Δδ15N and Δδ13C from aquatic systems, including three new field estimates. Carnivores had significantly higher Δδ15N values than herbivores. Furthermore, carnivores, invertebrates, and lab-derived estimates were significantly more variable than their counterparts ( f-test, p < 0.00001). Δδ13C was higher for carnivores than for herbivores (p = 0.001), while variances did not differ significantly. Excluding herbivores, the average Δδ15N and Δδ13C were 3.4‰ and 0.8‰, respectively. But even with unbiased fractionation estimates, there is variation in isotopic fractionation that contributes to error in quantitative isotope model outputs. We simulated the error variance in δ15N-based estimates of trophic position and two-source δ13C diet mixing models, explicitly considering the observed variation in Δδ15N and Δδ13C, along with the other potential error sources. The resultant error in trophic position and mixing model outputs was generally minor, provided that primary consumers were used as baseline indicators for estimating trophic position and that end member d13C values in dietary mixing models were sufficiently distinct.

Variation in δ15N and δ13C trophic fractionation: Implications for aquatic food web studies

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.

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

Stable isotope ratios provide clues about the origins and transformations of organic matter. A few key reactions control the isotopic composition of most organic matter. Isotopic variations introduced by these reactions are often passed on with little change so that isotopic measurements can indicate natural pathways and flows “downstream” from these key reactions. When chemical and metabolic processes scramble the information content of molecules, isotopic compositions are often preserved. This realization has prompted increasing use of stable isotope analyses as a tool for understanding complex ecological processes.

E. B. Sherr

Papers

δ13C Measurements as Indicators of Carbon Flow in Marine and Freshwater Ecosystems