Stable isotope ratios of a tropical marine predator: confounding effects of nutritional status during growth

Stable isotope analysis of carbon and nitrogen is frequently used to study the diets and foraging ecology of marine predators. However, isotopic values may also be affected by an individual’s nutritional status and associated physiological processes. Here, we use C and N stable isotopes in blood and feathers of blue-footed booby chicks at the Galápagos Islands to examine how isotopic values are related to body condition and growth rate, and to assess the consistency in the isotope ratios of individuals during growth. Size dimorphism in blue-footed boobies provided an additional opportunity to examine how isotope ratios differ between sexes in relation to body size and growth rate. There was no significant difference between sexes but both C and N stable isotopes were significantly negatively related to the body condition of chicks. These data were consistent with individual variation in physiological processes affecting fractionation, although we cannot rule out the possibility that they were also influenced to some extent by population-level variation in the stable isotope ratios of prey fed to chicks, for instance related to prey size, depth or lipid content. Our results highlight the need for methods that take proper account of confounding physiological factors in isotopic studies of foraging ecology and diet.     

Incorporating temporally dynamic baselines in isotopic mixing models

Stable isotopes (particularly C and N) are widely used to make inferences regarding food web structure and the phenology of consumer diet shifts, applications that require accurate isotopic characterization of trophic resources to avoid biased inferences of feeding relationships. For example, most isotope mixing models require that endmembers be adequately represented by a single probability distribution; yet, there is mounting evidence that the isotopic composition of aquatic organisms often used as mixing model endmembers can change over periods of weeks to months. A review of the literature indicated that the delta13C values of five aquatic primary consumer taxa, commonly used as proxies of carbon production sources (i.e., trophic baselines), express seasonally dynamic cycles characterized by an oscillation between summer maxima and winter minima. Based on these results, we built a dynamic baseline mixing model that allows a growing consumer to track temporal gradients in the isotopic baselines of a food web. Simulations showed that the ability of a consumer to maintain or approach isotopic equilibrium with its diet over a realistic growth season was strongly affected by both the rate of change of the isotopic baseline and equilibration rate of the consumer. In an empirical application, mixing models of varying complexity were used to estimate the relative contribution of benthic vs. pelagic carbon sources to nine species of juvenile fish in a fluvial lake of the St. Lawrence River system (Québec, Canada). Estimates of p (proportion of carbon derived from benthic sources) derived from a static mixing model indicated broad interspecific variation in trophic niche, ranging from complete benthivory to > 95% reliance on pelagic food webs. Output from the more realistic dynamic baseline mixing model increased estimated benthivory by an average of 36% among species. Taken together, our results demonstrate that failing to identify dynamic baselines when present, and (or) matching consumers with baseline taxa that possess substantially different equilibration rates can seriously bias interpretation of stable isotope data. Additionally, by providing a formalized framework that allows both resources and consumers to shift their isotopic value through time, our model demonstrates a feasible approach for incorporating temporally dynamic isotope conditions in trophic studies of higher consumers.     

Source identification for oil-based drill cuttings on the seabed based on stable carbon isotopes

This paper demonstrates how stable carbon isotopes can be utilized to identify the source of oil-based drill cuttings discharges and estimate the time of their deposition. The base-oil used in drilling mud systems may be biodegraded by seabed microflora. Effects of such processes make it difficult to identify sources of discharges based on the oil’s molecular structure. However, the stable isotope fingerprint of the base oil may vary for each batch of base-oil. Because carbon isotopic ratios in compounds with more than ten carbons are resistant to fractionation during biodegradation, this may be used as a tool to identify the source of drill cuttings discharges.     

Foraging ecology of leatherback sea turtles in the Western North Atlantic determined through multi-tissue stable isotope analyses

Leatherback turtles, Dermochelys coriacea, are highly migratory, spending most of their lives submerged or offshore where their feeding habits are difficult to observe. In order to elucidate the foraging ecology of leatherbacks off Massachusetts, USA, stable isotope analyses were performed on leatherback tissues and prey collected from 2005 to 2009. Stable isotope ratios of nitrogen and carbon were determined in whole blood, red blood cells, blood plasma, muscle, liver, and skin from adult male, female, and subadult leatherbacks. Isotopic values were analyzed by body size (curved carapace length) and grouped by sex, and groups were tested for dietary differences. Gelatinous zooplankton samples were collected from leatherback foraging grounds using surface dip nets and stratified net tows, and prey contribution to leatherback diet was estimated using a two-isotope Bayesian mixing model. Skin and whole blood δ¹³C values and red blood cell δ¹⁵N values were correlated with body size, while δ¹³C values of red blood cells, whole blood, and blood plasma differed by sex. Mixing model results suggest that leatherbacks foraging off Massachusetts primarily consume the scyphozoan jellyfishes, Cyanea capillata and Chrysaora quinquecirrha, and ctenophores, while a smaller proportion of their diet comes from holoplanktonic salps and sea butterflies (Cymbuliidae). Our results are consistent with historical observations of leatherback turtles feeding on scyphozoan prey in this region and offer new insight into size- and sex-related differences in leatherback diet.     

Trophic ecology of Grey-headed albatrosses from Marion Island, Southern Ocean: insights from stomach contents and diet tracers

During chick-rearing, albatrosses can alternate between long foraging trips that provide the main source of food for the adults and short foraging trips that they use to feed their young. This flexibility in foraging behaviour can lead to differences in diet composition between adults and chicks and implies that they may be vulnerable in different ways to food shortages. The trophic ecology of the Grey-headed albatross Thalassarche chrysostoma was investigated at the sub-Antarctic Prince Edward Islands during the chick-rearing period in April 2006 using a combination of approaches. Diets of adults and chicks were assessed using stable isotope ratios and fatty acid (FA) profiles of blood and/or stomach oils, in addition to stomach contents analysis. Fish from the family Macrouridae and cephalopods (particularly the onychoteuthid Kondakovia longimana) were the primary prey, whereas crustaceans (krill Euphausia superba) represented a smaller proportion of the stomach contents. Stomach oil FA profiles contained more monounsaturated FA than the profiles of plasma, which were richer in saturated FA and arachidonic acid (20:4n-6). There was also a distinct separation of adults from chicks, with higher levels of monounsaturates in chick plasma, and higher saturated FA levels (particularly 16:0) in the adult plasma. Stable carbon isotope ratios of whole blood were similar in adults and chicks, whereas stable nitrogen isotope ratios showed significant enrichment by >1‰ in chicks. The combined FA, stable isotopes and stomach contents analyses suggest clear differences in diet quality between adults and chicks, with chicks feeding at a higher trophic position through feeding more on highly nutritious fish and adults keeping much of the less nutritious zooplankton for themselves.     

Aquatic and terrestrial organic matter in the diet of stream consumers: implications for mercury bioaccumulation

The relative contribution of aquatic vs. terrestrial organic matter to the diet of consumers in fluvial environments and its effects on bioaccumulation of contaminants such as mercury (Hg) remain poorly understood. We used stable isotopes of carbon and nitrogen in a gradient approach (consumer isotope ratio vs. periphyton isotope ratio) across temperate streams that range in their pH to assess consumer reliance on aquatic (periphyton) vs. terrestrial (riparian vegetation) organic matter, and whether Hg concentrations in fish and their prey were related to these energy sources. Taxa varied in their use of the two sources, with grazing mayflies (Heptageniidae), predatory stoneflies (Perlidae), one species of water strider (Metrobates hesperius), and the fish blacknose dace (Rhinichthys atratulus) showing strong connections to aquatic sources, while Aquarius remigis water striders and brook trout (Salvelinus fontinalis) showed a weak link to in-stream production. The aquatic food source for consumers, periphyton, had higher Hg concentrations in low-pH waters, and pH was a much better predictor of Hg in predatory invertebrates that relied mainly on this food source vs. those that used terrestrial C. These findings suggest that stream biota relying mainly on dietary inputs from the riparian zone will be partially insulated from the effects of water chemistry on Hg availability. This has implications for the development of a whole-system understanding of nutrient and material cycling in streams, the choice of taxa in contaminant monitoring studies, and in understanding the fate of Hg in stream food webs.     

Coral cavity sponges depend on reef-derived food resources: stable isotope and fatty acid constraints

The diet of cavity sponges on the narrow fringing reefs of Curaçao, Caribbean was studied. The origin and resources of the bulk food of these sponges, i.e., dissolved organic matter (DOM), were identified using stable carbon and nitrogen isotopes and fatty acid biomarkers. We found that phytoplankton and its derived DOM from the adjacent open sea and from reef overlying water is not the main source of food for most of the sponges examined nor is bacterioplankton. Interestingly, dual stable isotope signatures (δ¹³C_org, δ¹⁵N_org) and fatty acid biomarkers appoint coral mucus and organic matter derived from crustose coralline algae (CCA) as probable food sources for encrusting sponges. Mucus-derived DOM may contribute up to 66% to the diet of examined sponges based on results of dual isotope mixing model analysis. The contribution of CCA (as purported representative for benthic algae) was smaller with values up to 31%. Together, mucus- and CCA-derived substrates contributed for 48–73% to the diet of sponges. The presence of the exogenous fatty acid 20:4ω6 in sponges, which is abundant in coral mucus of Madracis mirabilis and in CCA, highlights these reef-derived resources as sources of nutrition for DOM feeding cavity sponges. The relatively high concentrations of exogenous 20:4ω6 in all sponges examined supports our hypothesis that the bulk of the food of the cavity sponge community is reef-derived. Our results imply that cavity sponges play an important role in conserving food and energy produced within the reef.     

Differences in diet composition and foraging patterns between sexes of the Magellanic penguin (Spheniscus magellanicus) during the non-breeding period as revealed by δ13C and δ15N values in feathers and bone

In diving seabirds, sexual dimorphism in size often results in sex-related differences of foraging patterns. Previous research on Magellanic penguins, conducted during the breeding season, failed to reveal consistent differences between the sexes on foraging behavior, despite sexual dimorphism. In this paper, we tested the hypothesis that male and female Magellanic penguins differ in diet and foraging patterns during the non-breeding period when the constraints imposed by chick rearing activities vanish. We used stable isotope ratios of carbon and nitrogen in feather and bone to characterize the diet and foraging patterns of male and female penguins in the South Atlantic at the beginning of the 2009–2010 and 2010–2011 post-breeding seasons (feathers) and over several consecutive breeding and migratory seasons (bone). The mean δ¹³C and δ¹⁵N values of feathers showed no differences between the sexes in any of the three regions considered or in the diet composition between the sexes from identical breeding regions; however, Bayesian ellipses showed a higher isotopic niche width in males at the beginning of the post-breeding season. Stable isotope ratios in bone revealed the enrichment of males with δ¹³C compared with females across the three regions considered. Furthermore, the Bayesian ellipses were larger for males and encompassed those of females in two of the three regions analyzed. These results suggest a differential use of winter resources between the sexes, with males typically showing a larger diversity of foraging/migratory strategies. The results also show that dietary differences between male and female Magellanic penguins may occur once the constraints imposed by chick rearing activities cease at the beginning of the post-breeding season.     

The emergence of stable isotopes in environmental and forensic geochemistry studies: a review

In the past decade, environmental forensics has emerged as a discipline directed toward determining parties liable for causing spills of contaminants into the environment. Such investigations, while geared toward determining the guilty parties in order to recover costs of the cleanup and remediation, require various questions to be addressed. These include determination of the nature of the product; Where did it come from?; Extent of weathering, if any; How long has it been there?; and Is it degrading naturally? Traditionally, these studies have been addressed through utilization of techniques such as gas chromatography (GC) and gas chromatography–mass spectrometry (GCMS). However, in recent years, stable isotopes, primarily determined through the use of combined gas chromatography–isotope ratio mass spectrometry (GCIRMS), have emerged as an equally important tool in environmental forensics. For relatively low molecular, volatile compounds such as MTBE, BTEX, or chlorinated solvents, the isotopes, primarily carbon and hydrogen, have been used extensively for evaluating the onset of natural attenuation. For larger molecules such as PCBs or PAHs, in which the effects of biodegradation on the isotope composition of these molecules is minimal, the isotopic fingerprints of the individual compounds can be used for correlation purposes. In this paper, a brief introduction to isotope geochemistry will be given, followed by a review of applications of stable isotopes to a variety of environmental problems. While the review may not necessarily be exhaustive, it will provide a comprehensive overview of areas where isotopes have been used and potential applications for the future. Most of the review is concerned with carbon and hydrogen isotopes, although a brief overview of the emerging area of chlorine isotopes will also be discussed.     

Identifying oceanic foraging grounds of sea turtles in the Atlantic using lead isotopes

Many species of marine organisms go through ontogenetic shifts that occur in unknown or inaccessible locations. Finding these areas is crucial to understand connectivity and resilience of populations, both of which have conservation implications. When extrinsic markers are not suitable to track organisms, intrinsic markers can be useful to infer the location of inaccessible areas where these cryptic stages occur. Our study focuses on the location of oceanic foraging areas of the cryptic early juvenile stage of green turtles, Chelonia mydas, in the Atlantic. Due to the small size of hatchlings, the use of telemetry is limited to short periods of time and small spatial ranges, which do not allow determining the location of oceanic foraging areas. We used lead (Pb) stable isotopes to determine the possible location of oceanic foraging areas of small green turtles in the Atlantic Ocean. Pb stable isotope ratios in the scute tissue deposited when turtles were in the oceanic habitat were compared to ratios of major sources of lead in the Atlantic and oceanic areas in the Atlantic to determine the location of oceanic foraging grounds. The Pb isotope ratios in the scute of oceanic-stage green turtles indicated that turtles use different regions in the Atlantic and that they are capable of transatlantic migrations. We compare the oceanic locations identified by this study with those suggested by two previous studies.     

Evidence of abalone (<Emphasis Type="Italic">Haliotis rubra</Emphasis>) diet from combined fatty acid and stable isotope analyses

Abalone are common herbivores throughout temperate and tropical waters, and yet the contribution of red and brown macroalgae to the diet of wild abalone remains unclear. In the northern hemisphere, adult abalone are considered to consume predominantly brown algae, but in the southern hemisphere abalone are thought to prefer red algae. Conventional methods such as gut content analysis and feeding trials provide some insight into diet choice, but the associated biases of these techniques create uncertainty surrounding the aforementioned variability in abalone diet. We use combined stable isotope and fatty acid analysis to determine the relative contribution of red algae, brown algae and detritus/microalgae to the diet of wild abalone in Tasmanian waters. Stable isotopes of carbon suggest that brown algae and/detritus are a more important source of carbon than red algae. Fatty acid analysis confirmed the larger contribution of brown algae to the diet of abalone, and also identified the bacterial and diatom component of detritus to be an important contributor to abalone diet. These results show combined use of chemical tracers to be a promising technique for resolving abalone diet, and challenge current perceptions regarding spatial variability in abalone diet choice.     

Differences in stable isotopes in blood and feathers of seabirds are consistent across species, age and latitude: implications for food web studies

Stable isotopes of growing feathers and blood both represent assimilated diet, and both tissues are used to study the diet and foraging distribution of marine and terrestrial birds. Although most studies have assumed that both tissues represent a difference of one trophic level to diet, the enrichment factors of blood and feathers may differ, especially where endogenous reserves are used as precursors during feather synthesis. In this study, we compare carbon and nitrogen stable isotopes of blood and simultaneously growing feathers of five species of Procellariiformes, representing five genera, different geographical regions and different life stages (chicks and adults). In all species, feathers were enriched in ¹⁵N and ¹³C compared with blood. Isotopic values of carbon and nitrogen were correlated in different tissues growing simultaneously for most species analyzed, suggesting that mathematical corrections could be used to compare different tissues. Our results imply that more care needs to be taken when comparing stable isotope signatures across studies assuming different tissues are equivalent indicators of trophic ecology.     

Predator–prey models: an application for the plankton dynamics of lake Geneva

In this paper we present a hierarchical Bayesian analysis for a predator–prey model applied to ecology considering the use of Markov Chain Monte Carlo methods. We consider the introduction of a random effect in the model and the presence of a covariate vector. An application to ecology is considered using a data set related to the plankton dynamics of lake Geneva for the year 1990. We also discuss some aspects of discrimination of the proposed models.     

Seasonal variations in the stable carbon and nitrogen isotope ratios (13C/12C and 15N/14N) of primary producers and consumers in a western Mediterranean coastal lagoon

Stable carbon and nitrogen isotope ratios (¹³C/¹²C and ¹⁵N/¹⁴N) of primary producers and consumers were investigated seasonally throughout 1999, in order to describe the food web in a western Mediterranean coastal lagoon (Lake of Sabaudia, central Italy). Particulate organic matter and algal material (seagrass epiphytes and macroalgae) seem to constitute the main food sources for primary consumers (zooplankton and small benthic invertebrates, respectively) throughout the sampling year, while the seagrass Cymodocea nodosa appears to play a negligible trophic role. As regards the ichthyofauna, carbon stable isotopes differentiated between planktivore and benthivore fish species. However, a benthic-pelagic coupling seems to occur, with some fish of higher trophic levels feeding both on benthic and pelagic materials. Analysis of variance showed that the interaction between the three main factors (species2size2season) significantly affects the isotopic composition of fish, suggesting the presence of intra- and inter-specific resource partitioning. Wide seasonal variations in the isotopic composition were observed in organic matter sources, invertebrates and fish, with a general trend towards depleted values in winter and enriched values in summer. The winter depletion of organic matter sources may be due to several environmental factors and seems to be mirrored in the upper trophic levels. Primary producers and invertebrates are known to have shorter time-integrated isotopic signatures than vertebrates, yet fish also exhibited seasonal isotopic differences. We concluded that the examined fish species can assume a new muscle isotopic signature relatively quickly in response to changes in the isotopic composition of their diet and/or diet shifts.     

Terrestrial, benthic, and pelagic resource use in lakes: results from a three-isotope Bayesian mixing model

Fluxes of organic matter across habitat boundaries are common in food webs. These fluxes may strongly influence community dynamics, depending on the extent to which they are used by consumers. Yet understanding of basal resource use by consumers is limited, because describing trophic pathways in complex food webs is difficult. We quantified resource use for zooplankton, zoobenthos, and fishes in four low-productivity lakes, using a Bayesian mixing model and measurements of hydrogen, carbon, and nitrogen stable isotope ratios. Multiple sources of uncertainty were explicitly incorporated into the model. As a result, posterior estimates of resource use were often broad distributions; nevertheless, clear patterns were evident. Zooplankton relied on terrestrial and pelagic primary production, while zoobenthos and fishes relied on terrestrial and benthic primary production. Across all consumer groups terrestrial reliance tended to be higher, and benthic reliance lower, in lakes where light penetration was low due to inputs of terrestrial dissolved organic carbon. These results support and refine an emerging consensus that terrestrial and benthic support of lake food webs can be substantial, and they imply that changes in the relative availability of basal resources drive the strength of cross-habitat trophic connections.     

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.     

Identification and apportionment of sources of lead in human tissue

A brief review of the use of stable lead isotope ratio measurements to identify and apportion sources of lead in human tissue is followed by examples of the use of inductively coupled plasma source mass spectrometry for such studies. Inductively coupled plasma source mass spectrometry (ICP-MS) has only recently been used for measurements of coupled ratios in body tissues and fluids and in environmental sources of lead. Generally, the inaccuracy of these measurements is about −0.2% and the imprecision less than 0.5%. This analytical performance is sufficient to detect the much larger changes in²⁰⁶Pb:²⁰⁷Pb ratios of −2% or higher, seen in environmental lead exposure and in childhood lead poisoning. Measurements of lead isotope ratios by ICP-MS have been used to identify specific sources of childhood lead poisoning and to indicate the relative importance of environmental sources, such as drinking water and lead from petrol. Populations in the United Kingdom with low lead uptake usually have²⁰⁶:Pb²⁰⁷Pb ratios in body tissues within the range 1.13 ± 0.01. Significant deviations from this range have been seen in response to increased uptake from lead in: drinking water in parts of Scotland (source ratio ∼1.18 and petrol lead in inner London (source ratio ∼1.07). The dominant source for some Scottish subjects with high concentrations of lead in blood or in teeth was water, which contributed approximately 60% to body lead. Petrol lead was shown to be a significant contributor (30–40%) to the body lead of inner London children.     

Improved estimates of certainty in stable-isotope-based methods for tracking migratory animals.

The use of stable-hydrogen isotopes (deltaD) has become a common tool for estimating geographic patterns of movement in migratory animals. This method relies on broad and relatively predictable geographic patterning in deltaD values of precipitation, but these patterns are not estimated without error. In addition, deltaD measurements are relatively imprecise, particularly for organic tissue. Most models for estimating geographic locations have ignored these sources of error. Common modeling approaches include regression, range-matching, and likelihood-based assignment tests (including discriminant analysis). Here, we show the benefits of a simple stochastic extension to likelihood-based assignment tests that incorporates two estimable sources of error and describe the resulting influence on the certainty of assigning breeding origins for wintering American Redstarts (Setophaga ruticilla), a small Nearctic-Neotropical migratory bird. Through simulation, we incorporated both spatial interpolation error associated with models of deltaD in precipitation and analytical error associated with the measurement of deltaD in tissue samples. In general, assignments that did not include these sources of error fell within the ranges of the stochastic results, but the difference in proportion of birds assigned to any one breeding region varied by as much as 54%. To explore how the distribution of assignments generated from error models influenced the application of these results, we developed a simple model of winter habitat loss. We removed the proportion of Redstarts wintering at a particular site from the global population and then used the isotope-based assignments to predict the resulting population declines for each breeding region. This gave distributions of change in population sizes, some of which included no change or even a population increase. The sources of error we modeled may challenge the degree of certainty in the use of stable-isotope-based data on connectivity to predict population dynamics of migratory animals. We suggest that stronger inference will result from incorporating these sources of error into future studies that use deltaD or other stable isotopes to infer the geographic origin of individuals.     

A jellyfish diet for the herbivorous green turtle Chelonia mydas in the temperate SW Atlantic

Feeding ecology of juvenile green turtles (Chelonia mydas) was studied from 2008 to 2011 at Samborombón Bay (35°30′–36°30′S, Argentina), combining data on digestive tract examination and stable isotope analysis through a Bayesian mixing model. We found that animal matter, in particular gelatinous plankton, was consumed in large proportions compared to herbivorous food items such as terrestrial plants and macroalgae. This diet is facilitated by the high abundance of gelatinous plankton in the region, thus confirming the adaptive foraging behaviour of the juveniles according to prey abundance in the SW Atlantic. To our knowledge, this is the first study to employ this combination of techniques and to conclusively demonstrate that animal matter, in particular gelatinous plankton, is important in the diet of the neritic green sea turtles.     

Environmental correlates of large-scale spatial variation in the δ15N of marine animals

Marine Biology - Nitrogen stable isotopes can be used to estimate the trophic position of consumers in food webs. However, the nitrogen stable isotope ratios (δ15N) of primary producers at the...