Integrating stomach content and stable isotope analyses to quantify the diets of pygoscelid penguins

Stomach content analysis (SCA) and more recently stable isotope analysis (SIA) integrated with isotopic mixing models have become common methods for dietary studies and provide insight into the foraging ecology of seabirds. However, both methods have drawbacks and biases that may result in difficulties in quantifying inter-annual and species-specific differences in diets. We used these two methods to simultaneously quantify the chick-rearing diet of Chinstrap (Pygoscelis antarctica) and Gentoo (P. papua) penguins and highlight methods of integrating SCA data to increase accuracy of diet composition estimates using SIA. SCA biomass estimates were highly variable and underestimated the importance of soft-bodied prey such as fish. Two-source, isotopic mixing model predictions were less variable and identified inter-annual and species-specific differences in the relative amounts of fish and krill in penguin diets not readily apparent using SCA. In contrast, multi-source isotopic mixing models had difficulty estimating the dietary contribution of fish species occupying similar trophic levels without refinement using SCA-derived otolith data. Overall, our ability to track inter-annual and species-specific differences in penguin diets using SIA was enhanced by integrating SCA data to isotopic mixing modes in three ways: 1) selecting appropriate prey sources, 2) weighting combinations of isotopically similar prey in two-source mixing models and 3) refining predicted contributions of isotopically similar prey in multi-source models.     

Ontogenetic and sexual characterization of the feeding habits of franciscanas,Pontoporia blainvillei,based on tooth dentin carbon and nitrogen stable isotopes

We assessed the feeding habits of franciscanas (Pontoporia blainvillei) from southern Brazil through carbon and nitrogen stable isotopes measured in the tooth dentin of dolphins from different age classes. Generalized additive models were fitted to assess whether sex, age, and total body length explained changes in franciscanas isotope ratios. Bayesian isotope mixing models were applied to estimate the contributions of prey items, selected based on stomach content analysis (SCA), to franciscanas' diet. Different trophic enrichment factors (TEF) were applied and the results were compared using Bhattachayya's coefficient index to test for differences in model outputs and to define the best TEF for franciscanas. We found different relative importance of the main prey between the methods, and the contribution by squids could be lower than suggested by SCA. Because the dentin deposited during nursing seems to influence the isotopic ratios of juveniles, caution is recommended when interpreting dietary estimates based on whole tooth samples. This study is the first to characterize ontogenetic shifts in franciscanas' feeding habits using stable isotope analysis (SIA) and testing different TEF values. SIA and SCA complement each other in dietary studies, mitigating potential biases resulting from the use of each method separately.     

Gut content and stable isotope analyses provide complementary understanding of ontogenetic dietary shifts and trophic relationships among fishes in a tropical river

1. Despite widespread recognition of the role of body size in fish trophic ecology, little attention has been focused on this issue in isotopic studies, particularly in tropical systems. 2. We used analyses of stomach contents and stable isotopes to examine size‐related shifts in diet in a terapontid fish assemblage in the Australian wet–dry tropics. Stomach content analysis identified substantial ontogenetic dietary shifts in all species, corresponding to changes in body size–isotope trajectories for two species. Shifts away from relatively specialised diets of heavily ¹³C‐depleted insect larvae to consumption of a range of items across multiple basal carbon sources appeared to be the proximate cause of observed isotopic changes. 3. Allochthonous organic matter in the form of C3 riparian vegetation was particularly important to smaller terapontids before larger fish shifted to a broad range of dietary items and similarly broad range of basal carbon sources. 4. While there was general agreement between δ¹³C and stomach content analysis, there was minimal concurrence between the latter and δ¹⁵N isotopic derivation of estimates of trophic position. Due to factors such as omnivory, isotopically overlapping basal sources and uncertainties about rates of isotopic fractionation in both predator and prey species, stomach content analysis provides an essential complement to isotopic methodologies in tropical systems. 5. Given that basal sources supporting any individual species can change markedly with ontogeny, consideration of intraspecific, size‐related variation is necessary in isotopic studies of food web structure.     

The diet of great tit nestlings: Comparing observation records and stable isotope analyses

The diet of wild animals has been studied using many different strategies, approaches and methods in recent decades. In this regard, stable isotopes analysis (SIA) is becoming a widespread tool, but no study has yet, to our knowledge, compared diet estimations from SIA with direct observations of the diet of passerine nestlings. Accordingly, our aim was to test the predictive power of SIA for this purpose and identify potential confounding factors such as habitat effects. To do this, we compared isotopic signatures of δ¹³C and δ¹⁵N in the feathers of great tit (Parus major) nestlings, and the corresponding estimates of their diet based on stable isotope mixing models, with prey proportions delivered by their parents obtained through video-recordings. Between-nest differences in isotopic signatures of δ¹⁵N were larger than within-nest differences. We found that δ¹⁵N signatures of nestling feathers correlated positively with the proportion of spiders and negatively with the proportion of caterpillars in the nestlings’ diet, the most important prey types. On the other hand, between-nest and within-nest differences in δ¹³C ratios were of similar magnitude and δ¹³C ratios correlated mainly with the proportion of trees surrounding nest-boxes that were Quercus spp. Estimates of diet composition based on mixing models correlated with the observed nestling diet, yet effect sizes were quite low. Although mixing models are commonly used to ascertain diets, our data show that they can provide valuable information on the relative intake of prey types from different trophic levels; but when complex dietary patterns are recorded (e.g. due to the confounding effects of habitat and/or temporal variation) it can be difficult to draw firm conclusions about diet composition.     

Stable isotopes reveal Holocene changes in the diet of Adélie penguins in Northern Victoria Land (Ross Sea, Antarctica)

Adélie penguin (Pygoscelis adeliae) modern and fossil eggshells and guano samples collected from ornithogenic soils in Terra Nova Bay (Victoria Land, Ross Sea) were processed for carbon and nitrogen isotopic ratios with the aim of detecting past penguin dietary changes. A detailed and greatly expanded Adélie penguin dietary record dated back to 7,200 years BP has been reconstructed for the investigated area. Our data indicate a significant dietary shift between fish and krill, with a gradual decrease from past to present time in the proportion of fish compared to krill in Adélie penguin diet. From 7,200 to 2,000 years BP, δ¹³C and δ¹⁵N values indicate fish as the most eaten prey. The dietary contribution of lower-trophic prey in penguin diet started becoming evident not earlier than 2,000 years BP, when the δ¹³C values reveal a change in the penguin feeding behavior. Modern eggshell and guano samples reveal a major dietary contribution of krill but not a krill-dominated diet, since δ¹³C values remain much too high if krill prevail in the diet. According to the Holocene environmental background attested for Victoria Land, Adélie penguin dietary shifts between fish and krill seem to reflect penguin paleoecological responses to different paleoenvironmental settings with different conditions of sea-ice extension and persistence. Furthermore, Adélie penguin diet appears to be particularly affected by environmental changes in a very specific period within the breeding season, namely the egg-laying period when penguin dietary and feeding habit shifts are clearly documented by the δ¹³C of eggshell carbonate.     

Community structure affects trophic ontogeny in a predatory fish

While most studies have focused on the timing and nature of ontogenetic niche shifts, information is scarce about the effects of community structure on trophic ontogeny of top predators. We investigated how community structure affects ontogenetic niche shifts (i.e., relationships between body length, trophic position, and individual dietary specialization) of a predatory fish, brown trout (Salmo trutta). We used stable isotope and stomach content analyses to test how functional characteristics of lake fish community compositions (competition and prey availability) modulate niche shifts in terms of (i) piscivorous behavior, (ii) trophic position, and (iii) individual dietary specialization. Northern Scandinavian freshwater fish communities were used as a study system, including nine subarctic lakes with contrasting fish community configurations: (i) trout‐only systems, (ii) two‐species systems (brown trout and Arctic charr [Salvelinus alpinus] coexisting), and (iii) three‐species systems (brown trout, Arctic charr, and three‐spined sticklebacks [Gasterosteus aculeatus] coexisting). We expected that the presence of profitable small prey (stickleback) and mixed competitor–prey fish species (charr) supports early piscivory and high individual dietary specialization among trout in multispecies communities, whereas minor ontogenetic shifts were expected in trout‐only systems. From logistic regression models, the presence of a suitable prey fish species (stickleback) emerged as the principal variable determining the size at ontogenetic niche shifts. Generalized additive mixed models indicated that fish community structure shaped ontogenetic niche shifts in trout, with the strongest positive relationships between body length, trophic position, and individual dietary specialization being observed in three‐species communities. Our findings revealed that the presence of a small‐sized prey fish species (stickleback) rather than a mixed competitor–prey fish species (charr) was an important factor affecting the ontogenetic niche‐shift processes of trout. The study demonstrates that community structure may modulate the ontogenetic diet trajectories of and individual niche specialization within a top predator.     

Past and present trophic position and decadal changes in diet of Yellow-legged Gull in the Azores Archipelago, NE Atlantic

This study evaluates the trophic position of adult Yellow-legged Gulls Larus michahellis atlantis resident in the Azores archipelago in the past (1921–1928) and in the present (2009–2010), and analyses the decadal variation in the diet of breeding birds from the 1990s to the 2000s for three main colonies (Topo Islet, Baixo Islet and Mistério da Prainha). Using mixing models, we compared stable isotope signatures of nitrogen and carbon in adult breast feathers between birds from 1921 to 1928 (held in museum collections) and 2009 to 2010, jointly with both isotopic signatures of their main prey groups (fish, goose barnacles (Lepas anatifera), seabirds, mammals and refuse). The diet of breeding birds was analysed using pellets collected in 1989, 1995, 1996, 2004, 2009 and 2010. Stable isotopes analysis (SIA) results were in accordance with the results provided from the analysis of pellets, showing a relatively recent and significant change in the diet of adult gulls. In particular, SIA revealed a significant decrease in the trophic position of Yellow-legged Gulls in the Azores, over the last 89 years in response to the decrease in the consumption of seabirds and fish and, an increase in the consumption of marine invertebrates (goose barnacles) and refuse. The analysis of pellets confirmed the significant decrease in the fish ingested, whereas the ingestion of lower trophic level prey (i.e. goose barnacles, mammals and refuse) increased. Both methods reflect the feeding plasticity and opportunistic foraging behavior of this species, and are in accordance with patterns described for continental Europe.     

Using Bayesian stable isotope mixing models to estimate wolf diet in a multi-prey ecosystem

Stable isotope analysis (SIA) of wolf (Canis lupus) tissues can be used to estimate diet and intra-population diet variability when potential prey have distinct δ¹³C and δ¹⁵N values. We tested this technique using guard hairs collected from 44 wolves in 12 northwestern Montana packs, summer 2009. We used hierarchical Bayesian stable isotope mixing models to determine diet and scales of diet variation from δ¹³C and δ¹⁵N of wolves and potential prey, white-tailed deer (Odocoileus virginianus), mule deer (Odocoileus hemionus), elk (Cervus canadensis), moose (Alces alces), snowshoe hare (Lepus americanus), and other prey. As a check on SIA results, we conducted a separate diet analysis with temporally matched scats (i.e., collected in summer 2008) from 4 of the same packs. Wolves were centered on the ungulate prey in the isotope mixing space. Both methods revealed differences among pack diets and that wolves may consume moose in greater proportions than predicted by available biomass. Stable isotope analysis, and scat results were not entirely concordant; assumptions related to tissues of use in SIA, hair growth period in wolves, and scat sampling may have contributed to a mismatch between methods. Incorrect fractionation values, insufficient separation of prey in the isotope mixing space, choice of prior information in the Bayesian mixing models, and unexplained factors may have distorted diet estimates. However, the consistently high proportion of moose in pack diets suggests that increased population monitoring would benefit management of moose and wolves. Our results also support suggestions of other researchers that species-specific fractionation values should be used whenever possible, and that SIA may sometimes only provide indices of use for general groups of prey (e.g., large ungulates). © 2012 The Wildlife Society.     

Trophic behaviour of juvenile reef fishes inhabiting interlinked mangrove–seagrass habitats in offshore mangrove islets

Stable isotope (δ¹³C and δ¹⁵N) and gut content analyses were used to investigate size‐related feeding habits of four reef fishes (the beaugregory Stegastes leucostictus, the french grunt Haemulon flavolineatum, the schoolmaster snapper Lutjanus apodus and the yellowtail snapper Ocyurus chrysurus) inhabiting an offshore (non‐estuarine) mangrove islet off Belize, Central America. Comparisons of isotopic niche space and Schoener diet similarity index suggested a low to moderate degree of niche overlap between fish size groups. The δ¹³C gradient between mangrove and seagrass prey as well as results of Bayesian mixing models revealed that sampled fishes relied mostly on seagrass prey items. Only small and large juveniles of the carnivorous species L. apodus derived a part of their diet from mangroves by targeting mangrove‐associated Grapsidae crabs and fish prey, respectively. Isotopic niche shifts were particularly obvious for carnivorous fishes that ingested larger prey items (Xanthidae crabs and fishes) during their ontogeny. The utilization of mangrove food resources is less than expected and depends on the ecology and life history of the fish species considered. This research highlights that mangrove‐derived carbon contributed relatively little to the diets of four fish taxa from an offshore mangrove islet.     

Long‐term dietary shift and population decline of a pelagic seabird—A health check on the tropical Atlantic?

In the face of accelerating ecological change to the world's oceans, seabirds are some of the best bio‐indicators of marine ecosystem function. However, unravelling ecological changes that pre‐date modern monitoring programmes remains challenging. Using stable isotope analysis of feathers and regurgitants collected from sooty terns (Onychoprion fuscatus) nesting at a major Atlantic colony, we reconstructed a long‐term dietary time series from 1890 to the present day and show that a significant dietary shift occurred during the second half of the twentieth century coinciding with an apparent population collapse of approximately 84%. After correcting for the “Suess Effect,” δ¹³C in feathers declined by ~1.5‰ and δ¹⁵N by ~2‰ between the 1890s and the present day, indicating that birds changed their diets markedly over the period of population decline. Isotopic niches were equally wide before and after the population collapse but isotopic mixing models suggest that birds have grown ever more reliant on nutrient‐poor squid and invertebrates as teleost fish have declined in availability. Given that sooty terns rely heavily on associations with sub‐surface predators such as tuna to catch fish prey, the rapid expansion of industrialized fisheries for these species over the same period seems a plausible mechanism. Our results suggest that changes to marine ecosystems over the past 60 years have had a dramatic impact on the ecology of the most abundant seabird of tropical oceans, and highlight the potentially pervasive consequences of large predatory fish depletion on marine ecosystem function.     

Comparing pellet and stable isotope analyses of nestling Bonelli's Eagle Aquila fasciata diet

Diet analyses are central to the study of avian trophic ecology, and stable isotope analyses have made an increasing contribution in the last two decades. Few isotopic studies have assessed the diet of raptor species, which are more frequently analysed by conventional diet methods such as pellet analysis. In this study, we compare prey consumption estimates of nestling Bonelli's Eagles Aquila fasciata from conventional pellet analysis (in terms of items and biomass) and stable isotopic mixing models (SIAR) using δ¹³C, δ¹⁵N and δ³⁴S of feathers. The pellet analysis showed that European Rabbits Oryctolagus cuniculus, pigeons (mainly Common Wood Pigeons Columba palumbus and Domestic Pigeons Columba livia dom.), Red‐legged Partridges Alectoris rufa, passerines, Yellow‐legged Gulls Larus michahellis and Eurasian Red Squirrels Sciurus vulgaris were the main prey, so they were selected for diet reconstructions in SIAR. At the population level, mean prey consumption estimates were similar for pellets (both items and biomass) and SIAR. At the territory level, the weighted kappa statistic showed good ordinal scale agreement in main prey consumption between items or biomass and SIAR. Although the intraclass correlation coefficient showed poor method agreement when considering all prey in the same analysis, the intraclass correlation coefficients for each prey category showed significant agreement between pellets and SIAR when estimating the consumption of Rabbits, pigeons and Gulls, with lower agreement for passerines and Squirrels. Lastly, there was poor method agreement for estimates of Partridges. Our results suggest an overall agreement between the pellet analysis and SIAR when estimating nestling Bonelli's Eagle diet at both the population and, to a lesser extent, the territory level, supporting the usefulness of isotopic mixing models when identifying the terrestrial and marine components of raptor diets.     

Adélie Penguin Population Diet Monitoring by Analysis of Food DNA in Scats

The Adélie penguin is the most important animal currently used for ecosystem monitoring in the Southern Ocean. The diet of this species is generally studied by visual analysis of stomach contents; or ratios of isotopes of carbon and nitrogen incorporated into the penguin from its food. There are significant limitations to the information that can be gained from these methods. We evaluated population diet assessment by analysis of food DNA in scats as an alternative method for ecosystem monitoring with Adélie penguins as an indicator species. Scats were collected at four locations, three phases of the breeding cycle, and in four different years. A novel molecular diet assay and bioinformatics pipeline based on nuclear small subunit ribosomal RNA gene (SSU rDNA) sequencing was used to identify prey DNA in 389 scats. Analysis of the twelve population sample sets identified spatial and temporal dietary change in Adélie penguin population diet. Prey diversity was found to be greater than previously thought. Krill, fish, copepods and amphipods were the most important food groups, in general agreement with other Adélie penguin dietary studies based on hard part or stable isotope analysis. However, our DNA analysis estimated that a substantial portion of the diet was gelatinous groups such as jellyfish and comb jellies. A range of other prey not previously identified in the diet of this species were also discovered. The diverse prey identified by this DNA-based scat analysis confirms that the generalist feeding of Adélie penguins makes them a useful indicator species for prey community composition in the coastal zone of the Southern Ocean. Scat collection is a simple and non-invasive field sampling method that allows DNA-based estimation of prey community differences at many temporal and spatial scales and provides significant advantages over alternative diet analysis approaches.     

Seasonal variation in terrestrial resource subsidies influences trophic niche width and overlap in two aquatic snake species: a stable isotope approach

Quantifying diet is essential for understanding the functional role of species with regard to energy processing, transfer, and storage within ecosystems. Recently, variance structure in the stable isotope composition of consumer tissues has been touted as a robust tool for quantifying trophic niche width, a task that has previously proven difficult due to bias in direct dietary analyses and difficulties in integrating diet composition over time. We used carbon and nitrogen stable isotope analyses to examine trophic niche width of two sympatric aquatic snakes, banded watersnakes Nerodia fasciata and black swamp snakes Seminatrix pygaea inhabiting an isolated wetland where seasonal migrations of amphibian prey cause dramatic shifts in resource availability. Specifically, we characterized snake and prey isotope compositions through time, space, and ontogeny and examined isotope values in relation to prey availability and snake diets assessed by gut content analysis. We determined that prey cluster into functional groups based on similarity of isotopic composition and seasonal availability. Isotope variance structure indicated that the trophic niche width of the banded watersnake was broader (more generalist) than that of the black swamp snake. Banded watersnakes also exhibited seasonal variation in isotope composition, suggesting seasonal diet shifts that track amphibian prey availability. Conversely, black swamp snakes exhibited little seasonal variation but displayed strong ontogenetic shifts in carbon and nitrogen isotope composition that closely paralleled ontogenetic shifts in their primary prey, paedomorphic mole salamanders Ambystoma talpoideum. Although niche dimensions are often treated as static, our results demonstrate that seasonal shifts in niche dimensions can lead to changes in niche overlap between sympatric species. Such short‐term fluctuations in niche overlap can influence competitive interactions and consequently the composition and dynamics of communities and ecosystems.     

Isotopic discrimination between food and blood and feathers of captive penguins: implications for dietary studies in the wild

Using measurements of naturally occurring stable isotopes to reconstruct diets or source of feeding requires quantifying isotopic discrimination factors or the relationships between isotope ratios in food and in consumer tissues. Diet-tissue discrimination factors of carbon ((13)C/(12)C, or delta (13)C) and nitrogen ((15)N/(14)N, or delta (15)N) isotopes in whole blood and feathers, representing noninvasive sampling techniques, were examined using three species of captive penguins (king Aptenodytes patagonicus, gentoo Pygoscelis papua, and rockhopper Eudyptes chrysocome penguins) fed known diets. King and rockhopper penguins raised on a constant diet of herring and capelin, respectively, had tissues enriched in (15)N compared to fish, with discrimination factors being higher in feathers than in blood. These data, together with previous works, allowed us to calculate average discrimination factors for (15)N between whole lipid-free prey and blood and feathers of piscivorous birds; they amount to +2.7 per thousand and +4.2 per thousand, respectively. Both fish species were segregated by their delta (13)C and delta (15)N values, and importantly, lipid-free fish muscle tissue was consistently depleted in (13)C and enriched in (15)N compared to whole lipid-free fish. This finding has important implications because previous studies usually base dietary reconstructions on muscle of prey rather than on whole prey items consumed by the predator. We tested the effect of these differences using mass balance calculations to the quantification of food sources of gentoo penguins that had a mixed diet. Modeling indicated correct estimates when using the isotopic signature of whole fish (muscle) and the discrimination factors between whole fish (muscle) and penguin blood. Conversely, the use of isotopic signatures of muscle together with discrimination factors between whole fish and blood (or the reverse) leads to spurious estimates in food proportions. Consequently, great care must be taken in the choice of isotopic discrimination factors to apply to wild species for which no controlled experiments on captive individuals have been done. Finally, our results also indicate that there is no need to remove lipids before isotopic analysis of avian blood.     

Characterization of food web structure of the upper continental slope of the Celtic Sea highlighting the trophic ecology of five deep‐sea fishes

In marine ecosystems, the study of trophic relationships has extensively benefited from the development of stable isotope analyses (SIA) as dietary tracers. SIA are particularly useful in elucidating the structure of deep sea food webs given the constraints involved in obtaining gut‐content data from deep trawling. We used carbon and nitrogen stable isotope analyses and Stable Isotope Bayesian Ellipses in R (SIBER) and Stable Isotope Analysis in R (SIAR) routines, to determine the trophic ecology of five deep‐sea fishes from the upper continental slope of the Celtic Sea. SIA made it possible to deduce some general tendencies in food‐web structure and species trophic interactions and confirmed diet determined by gut‐content analysis for the same species, in other ecoregions. More specifically, mixing models revealed that the deep sea species considered are omnivorous and are able to feed on all the sampled taxa. Based on isotopic ratio, no clear differences in fish diet could be detected from one species to another except for rabbit fish, which has benthic affinities. Three species, blackbelly rosefish, greater forkbeard and softhead grenadier showed overlapping isotopic niches. This study is the first attempt to describe the trophic ecology of deep sea species on the Celtic Sea upper continental slope. In the context of the development of ecosystem integrated modeling approaches for managing fisheries in the Celtic sea, and considering the vulnerability of deep‐water species, improving the knowledge on the trophic ecology of these local species is of importance in order to allow their sustainable exploitation.     

Relevance of forage fish in the diet of Magellanic penguins breeding in northern Patagonia,Argentina

We quantified the trophic niche of Magellanic penguins (Spheniscus magellanicus) breeding and moulting in Golfo San Jorge, Argentina, through conventional stomach content and stable isotope analysis. A total of 112 adults were flushed during the early and late chick stages of 2011 and 2012 at Isla Vernacci Norte, and at least 15 prey taxa were found, including fishes, cephalopods, crustaceans and polychaetes. Overall, Argentine anchovy (Engraulis anchoita) showed the highest contribution in terms of importance by mass (68.1–85.3%, depending on chick stage and year), except for the old chick stage in 2011 when the shortfin squid (Illex argentinus) was the main prey consumed (56.0%). Based on carbon and nitrogen isotopic values from a total of 256 blood samples, corresponding to young and old chicks and to adults of both sexes sampled throughout the incubation, chick and moult stages at the above mentioned colony and years, Bayesian mixing model outputs showed that Argentine anchovy was always the main prey (48–86%). Bayesian mixing model outputs obtained from adults of both sexes and their chicks during the late chick stage of 2013 at Isla Vernacci Norte, Isla Tova and Isla Leones also showed that Argentine anchovy was the main prey consumed. This is the first comprehensive assessment of Magellanic penguin diet composition in northern Patagonia, quantifying the relative contribution of prey in the diet of adults and chicks at different stages of the annual cycle and years, and confirms the relevance of a forage fish such as the Argentine anchovy in its trophic ecology.     

Seasonal and annual differences in the foraging ecology of two gull species breeding in sympatry and their use of fishery discards

Niche segregation between similar species will result from an avoidance of competition but also from environmental variability, including nowadays anthropogenic activities. Gulls are among the seabirds with greater behavioural plasticity, being highly opportunistic and feeding on a wide range of prey, mostly from anthropogenic origin. Here, we analysed blood and feather stable isotopes combined with pellet analysis to investigate niche partitioning between Audouin's gull Larus audouinii and yellow‐legged gull Larus michahellis breeding in sympatry at Deserta Island, southern Portugal, during 2014 and 2015. During the breeding season there was considerable overlap in the adults’ diet, as their stable isotope values of blood and primary feather (P1) did not differ, and their pellets were comprised mainly by marine fish species. However, Audouin's gulls presented higher occurrences of pelagic fish, while yellow‐legged gulls fed more on demersal fish, insects, and refuse. SIAR mixing models also estimated a higher proportion of demersal fish in the diet of yellow‐legged gulls. We also found differences between the two gull species in chicks’ feathers, suggesting that Audouin's gull adults selected prey with lower carbon isotope values to feed their young. Secondary feather (S8) of Audouin's gull presented higher isotope values compared to yellow‐legged gulls, indicating different foraging areas (δ¹³C) and/ or trophic levels (δ¹⁵N) between the two species in the non‐breeding season. During both the all‐year and non‐breeding periods the yellow‐legged gull showed a broader isotopic niche width than Audouin's gull in 2013, and in 2014 the two gull species exhibited different isotopic niche spaces. Our study suggests that both gull species foraged in association with fisheries during the breeding season. In this sense, a discard ban implemented under the new European Union Common Fisheries Policy may lead to a food shortage, therefore future research should closely monitor the population dynamics of Audouin's and yellow‐legged gulls.     

Stable isotope evidence of trophic interactions between introduced brook trout Salvelinus fontinalis and native brown trout Salmo trutta in a mountain stream of south-west France

The potential trophic impact of introduced brook trout Salvelinus fontinalis on native brown trout Salmo trutta in a mountain stream (south-west France) was investigated using stable isotope analysis (SIA). The isotopic signatures (δ¹³C and δ¹⁵N) of S. fontinalis were similar regardless of the absence or presence of S. trutta , and SIA mixing models revealed that S. fontinalis diet consisted mainly of terrestrial invertebrates. Conversely, a significant shift in S. trutta isotopic signatures (depletion of 1·6‰δ¹³C and enrichment of 0·6‰δ¹⁵N) was observed in sympatry with S. fontinalis ; this may be due to a dietary shift towards terrestrial invertebrates. Contrary to an expected dietary divergence in sympatry, an elevated level of dietary overlap was observed between the non-native and native salmonids when in co-occurrence. This dietary convergence is more likely to be due to behavioural interactions than to variations in food availability or fish displacements.     

Blood fatty acids indicate inter- and intra-annual variation in the diet of Adélie penguins: Comparison with stomach content and stable isotope analysis

Adélie penguin (Pygoscelis adeliae) diet is an important indicator of prevailing environmental conditions and resource availability. In this study, dietary variation within and between years was studied with fatty acid signature analysis (FASA), stomach content analysis (SCA) and stable isotope analysis (SIA). We profiled the fatty acid (FA) composition of whole blood collected from adult penguins throughout the breeding season, and from chicks during the crèche period, in 2001 and 2002. Differences were detected in FA profiles between years, breeding stage and age (adults vs. chicks). These patterns broadly corresponded to those observed from SCA and SIA, with a mix of krill and fish consumed in the early part of the breeding season in both years, krill dominating the diet during the chick-rearing periods in 2001, and fish in 2002. Different metabolic and physiological demands between stages, and ages, may also influence FA profiles but warrants further investigation. In-situ calibrations of adult FA blood profiles were made using corresponding stomach samples to quantify diet composition. Using linear discriminate function analysis, we classified adult FA profiles into 3 meal-types: krill, fish or mixed. A higher proportion of adults had fish-like profiles during the arrival and guard periods. Krill-like profiles dominated during the incubation and crèche periods, although there were a relatively high proportion of fish-like and mixed profiles as well. These patterns corresponded to results from SCA and SIA. This study demonstrates that FASA has the potential to be integrated with other dietary tools to enhance diet monitoring studies, which are currently integral to ecosystem management and conservation measures. The in-situ calibration method used offers a simple and effective alternative to more rigorous calibration techniques developed elsewhere.     

How important are seabirds in the diet of black rats on islands with a superpredator?

This study assessed the impact of introduced black rats (Rattus rattus) on Cory's shearwater (Calonectris diomedea borealis) in a multi-invaded insular ecosystem where rats are mesopredators. We hypothesized that black rats should have little impact on Cory's shearwaters in the presence of cats as superpredators. Stomach contents and stable isotope analysis (SIA) in tissues of black rats were analyzed to assess the trophic ecology and the importance of Cory's shearwater in their diet. We also studied the isotopic signature in tissues of house mouse (Mus domesticus) to confirm previous data showing no predation of this species on Cory's shearwaters. For both rodent species, temporal variation in diet composition in response to the availability of seabird prey was evaluated, and short- and long-term consistency in diet was tested using different tissues from the same individual. For black rats a Bayesian isotope mixing model (SIAR) was applied to determine the relative contribution of each prey to the individual diet. SIA of mouse tissues varied between the Cory's shearwater breeding and non-breeding periods. However, no significant differences were found in diet and SIA for black rats. In contrast, individuals of both species showed a strong consistency in diet which apparently benefited their body condition index. Although black rats supplement their diet with Cory's shearwater eggs and chicks (8.3% in stomach contents and 10.6% in the SIAR model), their current impact on the Cory's shearwater population appears to be small, probably due to several factors including the small size of the rat population and a high level of rat predation by cats.