Diet-tissue Discrimination Research Articles

Diet–tissue discrimination factors and turnover rates of carbon and nitrogen stable isotopes in the mottled skate Beringraja pulchra based on diet-switching experiments

Stable isotope analysis is robust tool to elucidate the diet, trophic position, and migration of typically large, highly mobile marine predators such as elasmobranchs. However, it is necessary to understand the diet–tissue discrimination factors and isotopic turnover rates of target animals. In this study, diet-switching experiments were conducted on the mottled skate (Beringraja pulchra (Liu, 1932)), caught off Sarufutsu, Hokkaido, Japan, to elucidate the diet–tissue discrimination factors and isotopic turnover rates for carbon and nitrogen stable isotopes. Ninety juvenile skates were fed Euphausia superba (C:N ratio = 3.9 ± 0.06, δ15N = 3.7±0.26‰ and δ13C = –25.9±0.15‰) to satiation at intervals of 2–3 days, and their muscle and liver tissues were serially sampled during 400 experimental days and used for stable isotope analysis. Both carbon and nitrogen stable isotopes reached steady state within the experimental days, and the estimated diet–tissue discrimination factors based on the fitting models were Δ13C = 1.5‰ and Δ15N = 2.3‰ for liver tissues and Δ13C = 3.4‰ and Δ15N = 3.5‰ for muscle tissues. The estimated turnover rates were 0.034 day–1 for δ13C and 0.039 day–1 for δ15N for liver tissues and were higher than those for muscle tissues (0.021 day–1 for δ13C and 0.019 day–1 for δ15N). In addition, the growth-based isotopic turnover rate model comprising juvenile skates revealed that the contribution of growth rate to the isotopic turnover rate was low.

Diet-tissue discrimination factors of three neotropical freshwater fishes and a comparison of the trophic position

The trophic discrimination factor (TDF) is a key parameter for stable isotope analysis and due to a lack of species-specific TDFs, mean universal values have been used, resulting in uncertainties about the trophic position of species and a call for more experiments. In this study, we have addressed the lack of experimental species-specific TDFs conducting three experiments of 128 days each to determine the TDF (muscle and liver) of three species, the piscivore Pseudoplatystoma corruscans (Spix & Agassiz, 1829), and the omnivores Piaractus mesopotamicus (Holmberg, 1887) and Astyanax lacustris (Lütken, 1875), tropical fishes native to the La Plata River basin. Then, we calculated the trophic position (TP) using the mean universal TDF from literature and the species-specific TDF produced in this study for Pseudoplatystoma corruscans. We estimated the TDFs for the three species through experiment and the values found differed from the mean universal TDF in the literature. Moreover, the TP was lower when using the species-specific TDFs. The TP is important for several analyses, including its use in functional diversity. Therefore, we recommend using species-specific TDF values for calculating TP once it differs from the results calculated with mean universal TDF.

Chemolithoautotrophic Organic Matter Contributions to Subterranean Food Webs Dominated by Filter-feeders

Chemolithoautotrophic Organic Matter Contributions to Subterranean Food Webs Dominated by Filter-feeders

Turnover Rates and Diet-Tissue Discrimination Factors of Nitrogen and Carbon Stable Isotopes in Seahorse Hippocampus reidi Juveniles Following a Laboratory Diet Shift.

Simple SummaryThe main aim of the present study was to ascertain the effect of two feeding schedules (including copepods and Artemia nauplii) on the early development and physiology of seahorse Hippocampus reidi juveniles. For that, we analyzed seahorse performance (growth and survival) and trophic patterns by means of stable isotopes. Our results highlight that the welfare and condition of juveniles were enhanced by extending the period of feeding on copepods up to day 10 after the male’s pouch release. The analysis of turnover rates for δ13C and δ15N revealed that switching copepods to Artemia nauplii at earlier developmental stages would reduce prey assimilation resulting in lower growth rates and survivals. The present study also provides for the diet–tissue discrimination factors for δ13C and δ15N in seahorse juveniles for the first time.The initial development of seahorse juveniles is characterized by low digestion capabilities. Stable isotope analysis is an effective tool in studies of trophic food webs and animal feeding patterns. The present study provides new insights for the understanding of growth and food assimilation in early developing seahorses following a laboratory diet switch. The study was performed in the early life stages of the seahorse Hippocampus reidi by assessing the influence of diet shift on changes and turnovers in carbon (δ13C) and nitrogen (δ15N) stable isotope in juveniles. Newborn seahorses were fed for 60 days following two feeding schedules (A6 and A11) based initially on copepods Acartia tonsa and subsequently on Artemia nauplii (since days 6 and 11, respectively). After the prey shift, we determined δ13C and δ15N turnover rates as functions of change in either body mass (fitting model G) and days of development (fitting model D), contributions of metabolism and growth to those turnover rates, and diet–tissue discrimination factors. Survival, final dry weight, and final standard length for diet A11 were higher compared to diet A6. The shift from copepods to Artemia led to fast initial enrichments in δ13C and δ15N. Afterwards, the enrichment was gradually reduced until the isotopic equilibrium with the diet was reached. In most cases, both fitting models performed similarly. The isotopic analysis revealed that 100% of tissue turnover was attributed to growth in diet A11, whereas 19–25% was linked to metabolism in diet A6. Diet–tissue discrimination factors were estimated for the first time in seahorse juveniles, resulting in higher estimates for diet A11 (2.9 ± 0.7‰ for δ13C; 2.5 ± 0.2‰ for δ15N) than in diet A6 (1.8 ± 0.1‰ for δ13C; 1.9 ± 0.1‰ for δ15N). This study highlights the relevance of feeding on copepods and their effect on isotopic patterns and discrimination factors in seahorse juveniles after a dietary shift. Regarding the application of the results achieved in relation to the feeding schedules in the rearing of H. reidi, a long period of feeding on copepods during the first days of development is highly recommended.

Four approaches for estimating isotope discrimination factors produce contrasting dietary estimates for bears

Isotope diet-tissue discrimination factors can significantly influence dietary proportions estimated with stable isotope mixing models. For bears, for which discrimination factors from controlled dietary experiments are currently lacking for most tissues and diets, literature reports 3 approaches have so far been used to compile carbon and nitrogen discrimination factors, and recently one additional approach has been proposed. We used these 4 approaches to compile carbon and nitrogen discrimination factors for the brown bear (Ursus arctos) population in Slovenia (southeastern Europe). By performing a sensitivity analysis of the stable isotope mixing models, we then tested how these different discrimination factors affected dietary estimates for bears. We found substantial differences in the dietary estimates among the applied approaches, with the approach that uses discrimination factors from different species producing most distinct results. Our results suggested that these differences were more pronounced among approaches with larger differences in discrimination factors and for isotopically less distinct food sources. We show that the 4 approaches can lead to contrasting conclusions about bear diets, which can have important ecological and management consequences. Therefore, when using stable isotope mixing models without experimentally obtained species-, diet-, and tissue-specific discrimination factors, the choice of the discrimination-factor compilation approach is vital. All 4 approaches have limitations that must be considered because they can result in erroneous discrimination factors when certain species, diets, and tissues are used. We review these limitations and provide recommendations about the use of these approaches in stable isotope dietary studies for bears.

African wild dogs (Lycaon pictus) show differences in diet composition across landscape types in Kruger National Park, South Africa

Abstract The Kruger National Park (KNP) is home to the last genetically viable, minimally managed population of African wild dogs (Lycaon pictus, wild dogs) in South Africa. Until 2004, this population remained stable, but since has been declining. In this study, we aimed to improve our understanding of the ecology of KNP wild dogs by estimating the relative contribution of different prey types to their diet across landscape types. Based on a Bayesian mixing model, we assessed wild dog diet and foraging preferences using stable isotope analysis. We sampled 73 individuals from 40 packs found in six different landscape types. In thickets, packs predominantly prey on small browsing and mixed-feeding species (accounting for ~73% of their diet), but occasionally hunt large grazers (~24%) and large browsers (~3%). In open landscape types where lions (Panthera leo) are more or less absent, such as in the Lowveld sour bushveld, wild dogs prey on large browsers and large grazers (~67%). Our results demonstrate that KNP wild dogs occupy a broader ecological niche than previously thought, with small browsers forming an integral part of their diet. We also present the first data describing differences in wild dog diet–tissue discrimination factors for tail hair and whiskers compared to respective stable nitrogen (δ15N) and carbon (δ13C) values obtained from feces of captive wild dogs, as well as from those of South Africa’s broader managed metapopulation. While these data should be considered preliminary, we suggest that until wild dog diet–tissue discrimination factors are calculated through a controlled feeding study, the discrimination factors calculated for the gray wolf (Canis lupus) should be used for wild dog-related isotope studies, rather than the often cited values for red foxes (Vulpes vulpes).

European perch (Perca fluviatilis) fed dietary insect meal (Tenebrio molitor): From a stable isotope perspective

Stable isotope analysis was conducted to investigate stable isotope ratios of carbon (δ13C) and nitrogen (δ15N), diet-tissue discrimination factors of carbon (Δ13C) and nitrogen (Δ15Ν). Bayesian mixing models were performed to assess relative contribution of insect meal and other ingredients to the development of tissues of European perch (Perca fluviatilis). Accordingly, four experimental formulations, characterized by the increasing inclusion levels of yellow mealworm (Tenebrio molitor) larvae meal (TM) at 0, 6.8, 13.5 and 20.3% as replacement for fishmeal at 0 (TM0), 25 (TM25), 50 (TM50) and 75% (TM75), respectively, were fed to juvenile perch (initial bodyweight, 20.81 ± 3.36 g) in a recirculated aquaculture system for 105 days.δ13C and δ15N of TM were -16.75 and 3.53‰ and significantly distinguished from other terrestrial and marine feed components (P < 0.05). Inclusion of dietary TM did not affect Δ13C value in blood and liver (P > 0.05) but did reduce in muscle (P < 0.05), whereas Δ15Ν was significantly increased with the increasing inclusion level of TM in all tissues (P < 0.05). The growth of perch had a significant negative relationship with diet-muscle Δ15Ν. The contribution of TM to muscle (7.7 ± 3.8%) was comparable to its dietary inclusion (6.8%) in TM25 but double in the blood (13 ± 6%). TM appeared to be an essential ingredient incorporated into liver, as its contribution was consistent or higher than dietary inclusion (TM25: 25.4 ± 12.1 vs. 6.8%; TM50: 31.1 ± 14.9 vs. 13.5%; and TM75: 29.4 ± 14.4 vs. 20.3%). The higher inclusion levels of TM (more than 6.8%) did not elevate its contribution to muscle, blood, and liver (probability, PBIC < 0.95) but significantly decreased that of fishmeal in all tissues (PBIC > 0.95). Soy-derived ingredients, soybean meal and soy protein, were an important ingredient in the development of all tissues regardless of dietary TM.The present study provided insightful information on the role of various diet components in perch tissues, which could underlie further development of aquafeed formulations for emerging perch farming in Europe.

Telemetry‐validated nitrogen stable isotope clocks identify ocean‐to‐estuarine habitat shifts in mobile organisms

Abstract Throughout their life history, many animals transition among heterogeneous environments to facilitate behaviours such as reproduction, foraging and predator avoidance. The dynamic environmental and biological conditions experienced by mobile species are integrated in the chemical composition of their tissues, providing retrospective insight into movement. Here, we present a unique application of nitrogen stable isotope clocks (‘isotopic clocks’), which integrate tissue turnover rates, consumer stable isotope ratios and habitat‐specific isotope baselines to predict time‐since‐immigration and the timing of habitat shifts in a migratory species. Nitrogen stable isotope values of blood plasma collected from juvenile sand tiger sharks Carcharias taurus, a species known to undertake seasonal movements between ocean and estuarine environments, were used to derive estimates of time‐since‐immigration and the timing of seasonal habitat shifts undertaken by this species. Nitrogen isotopic clocks estimated for 65 juvenile sand tiger sharks sampled across 6 years indicated that individual sharks predominantly arrived to estuarine habitats between June and July, with some individuals arriving as early as mid‐May. These estimates were validated by comparing isotope‐derived estuarine arrival times with those from acoustically tracked individuals. The median estuarine arrival day estimates from our isotopic approach aligned with estimates from acoustic telemetry for each sampling population. Sensitivity analyses indicated that isotopically inferred time‐since‐immigration and estuarine arrival estimates were robust to variation in isotopic turnover rate and diet tissue discrimination factors under multiple modelling scenarios. This suggests that parameterization of the nitrogen isotopic clock provides reliable estimates of time‐since‐immigration and day of arrival into new habitats if isotopic variation exists between origin and new locations. Our study presents a unique application of telemetry‐validated isotope clocks to derive retrospective estimates of time‐since‐immigration and timing of habitat shifts for animals that seasonally traverse heterogeneous environments. This approach can be readily applied across many temporal and spatial scales, and to other species and ecosystems, to facilitate rapid assessment of changes in animal habitat use and broader ecosystem structure.

Stable isotope turnover rates and fractionation in captive California yellowtail (Seriola dorsalis): insights for application to field studies

Stable isotope analysis (SIA) measurements from long-term captivity studies provide required parameters for interpretation of consumer SIA data. We raised young-of-the-year (14–19 cm) California yellowtail (Seriola dorsalis) on a low δ15N and δ13C diet (pellet aquaculture feed) for 525 days, then switched to a high δ15N and δ13C diet (mackerel and squid) for 753 days. Yellowtail muscle was sequentially sampled from each individual after the diet switch (0 to 753 days) and analyzed for δ15N and δ13C, allowing for calculation of diet-tissue discrimination factors (DTDFs) from two isotopically different diets (low δ15N and δ13C: pellets; high δ15N and δ13C: fish/squid) and turnover rates of 15N and 13C. DTDFs were diet dependent: Δ15N = 5.1‰, Δ13C = 3.6‰ for pellets and Δ15N = 2.6‰, Δ13C = 1.3‰ for fish/squid. Half-life estimates from 15N and 13C turnover rates for pooled yellowtail were 181 days and 341 days, respectively, but varied considerably by individual (15N: 99–239 d; 13C: 158–899 d). Quantifying DTDFs supports isotopic approaches to field data that assume isotopic steady-state conditions (e.g., mixing models for diet reconstruction). Characterizing and quantifying turnover rates allow for estimates of diet/habitat shifts and “isotopic clock” approaches, and observed inter-individual variability suggests the need for large datasets in field studies. We provide diet-dependent DTDFs and growth effects on turnover rates, and associated error around these parameters, for application to field-collected SIA data from other large teleosts.

From diet to hair and blood: empirical estimation of discrimination factors for C and N stable isotopes in five terrestrial mammals

AbstractCarbon and nitrogen stable isotope ratios are used widely to describe wildlife animal diet composition and trophic interactions. To reconstruct consumer diet, the isotopic differences between consumers and their diet items—called the trophic discrimination factor (TDF)—must be known. Proxies of diet composition are sensitive to the accuracy of TDFs. However, specific TDFs are still missing for many species and tissues because only a few controlled studies have been carried out on captive animals. The aim of this study was to estimate TDFs for hair and blood for carbon and nitrogen stable isotopes for caribou, moose, white-tailed deer, eastern coyote, and black bear. We obtained stable isotope ratios for diet items, hair, and blood samples, of 21 captive adult mammals. Diet–tissue discrimination factors for carbon in hair (∆ 13CLE) ranged from 0.96‰ to 3.72‰ for cervids, 3.01‰ to 3.76‰ for coyote, and 5.15‰ to 6.35‰ for black bear, while nitrogen discrimination factors (∆ 15N) ranged from 2.58‰ to 5.95‰ for cervids, 2.90‰ to 3.13‰ for coyote, and 4.48‰ to 5.44‰ for black bear. The ∆ 13CLE values in coyote blood components ranged from 2.20‰ to 2.69‰ while ∆ 15N ranged from 3.30‰ to 4.41‰. In caribou serum, ∆ 13CLE reached 3.34 ± 1.28‰ while ∆ 15N reached 5.02 ± 0.07‰. The TDFs calculated in this study will allow the evaluation of diet composition and trophic relationships between these five mammal species and will have important implications for the study of endangered caribou populations for which the use of noninvasive tissue sampling is highly relevant.

Exploring source differences on diet-tissue discrimination factors in the analysis of stable isotope mixing models

Stable isotope mixing models are regularly used to provide probabilistic estimates of source contributions to dietary mixtures. Whilst Bayesian implementations of isotope mixing models have become prominent, the use of appropriate diet-tissue discrimination factors (DTDFs) remains as the least resolved aspect. The DTDFs are critical in providing accurate inferences from these models. Using both simulated and laboratory-based experimental data, this study provides conceptual and practical applications of isotope mixing models by exploring the role of DTDFs. The experimental study used Mozambique Tilapia Oreochromis mossambicus, a freshwater fish, to explore multi-tissue variations in isotopic incorporation patterns, and to evaluate isotope mixing model outputs based on the experiment- and literature-based DTDFs. Isotope incorporation patterns were variable for both muscle and fin tissues among the consumer groups that fed diet sources with different stable isotope values. Application of literature-based DTDFs in isotope mixing models consistently underestimated the dietary proportions of all single-source consumer groups. In contrast, application of diet-specific DTDFs provided better dietary estimates for single-source consumer groups. Variations in the proportional contributions of the individual sources were, nevertheless, observed for the mixed-source consumer group, which suggests that isotope assimilation of the individual food sources may have been influenced by other underlying physiological processes. This study provides evidence that stable isotope values from different diet sources exhibit large variations as they become incorporated into consumer tissues. This suggests that the application of isotope mixing models requires consideration of several aspects such as diet type and the associated biological processes that may influence DTDFs.

Isotopic Discrimination (δ15N, δ13C) in Captive and Wild Common Murres (Uria aalge) and Atlantic Puffins (Fratercula arctica).

Studying the diet of consumers using stable isotopes provides insight into the foraging ecology of individuals and species. To accurately reconstruct the integrated diet of animals using stable isotope values, we must quantify diet-tissue discrimination factors (DTDFs), or the way in which stable isotopes in prey are incorporated into the tissues of consumers. To quantify DTDFs, controlled experiments are needed, whereby consumers are fed a constant diet. However, relatively few controlled-diet studies have been conducted for seabirds. In this study, captive adult Atlantic puffins (Fratercula arctica) and common murres (Uria aalge) were fed a two-source diet of capelin (Mallotus villosus) and Atlantic silverside (Menidia menidia) to determine the DTDFs for the cellular component of blood and plasma for both δ15N and δ13C. The DTDFs for the cellular component (Δ15N: ; Δ13C: ) and plasma (Δ15N: ; Δ13C: ) of puffins were similar to those for the cellular component (Δ15N: ; Δ13C: ) and plasma (Δ15N: ; Δ13C: ) of murres. We reconstructed the diet of wild murres and puffins breeding on the northeastern coast of Newfoundland using previously published DTDFs and estimated DTDFs from our feeding experiment. Reconstructed dietary proportions supported a priori knowledge of diet, although outputs were sensitive to the DTDF used. Despite the similarity of our DTDFs for puffins and murres, along with the similarity of our DTDFs with those of other seabird species, our sensitivity analysis revealed considerable differences among resultant dietary contributions from mixing models, further highlighting the importance of using species- and tissue-specific DTDFs to enhance knowledge in the foraging ecology of seabirds using stable isotopes.

Isotopic discrimination between carrion and elytra clippings of lab-reared American burying beetles (Nicrophorus americanus): Implications for conservation and evaluation of feeding relationships in the wild.

Differences in stable isotope composition between an animal and its diet are quantified by experimentally derived diet-tissue discrimination factors. Appropriate discrimination factors between consumers and prey are essential for interpreting stable isotope patterns in ecological studies. While available for many taxa, these values are rarely estimated for organisms within the carrion food web. We used a controlled-diet stable isotope feeding trial to quantify isotopic diet-tissue discrimination factors of carbon (δ13 C values) and nitrogen (δ15 N values) from laboratory-reared Nicrophorus americanus raised on carrion. We used exoskeleton samples of beetle elytra (wing covers) to determine diet-tissue discrimination factors using a continuous flow isotope ratio mass spectrometer equipped with an elemental analyzer. We also measured the isotopic compositions of five species of co-occurring, wild-caught burying beetles and evaluated feeding relationships. We found differences in stable carbon discrimination between carrion sources (mammalian and avian) and lab-reared beetles, but no difference in stable nitrogen discrimination. Values for δ13 C did not differ among wild-caught burying beetle species, but values for δ15 N were significantly different for the three species with overlapping breeding seasons. Furthermore, wild-caught burying beetles within our study area do not appear to use avian carrion resources to rear their young. This study informs future interpretation of stable isotope data for insects within the carrion food web. In addition, these results provide insight into carrion resources used by co-occurring burying beetle species in situ. We also demonstrated that independent of adult food type, the larval food source has a significant impact on the isotopic signatures of adult beetles, which can be estimated using a minimally invasive elytra clipping.

Assessing trophic position quantification methods for three piscivorous freshwater fish using stable isotopes and stomach contents

Accurate trophic position (TP) estimates are important for the development of ecosystem-based management plans. TPs can be quantified by carbon (δ13C) and nitrogen (δ15N) stable isotopes in tissues, but these can disagree with observed and perceived feeding ecology. A recent method that has used a scaled diet-tissue discrimination factor (DTDF), reflecting the inverse relationship between DTDF and δ15N, was found to better describe TPs of predatory fish species in marine ecosystems, but this has not been tested in freshwater ecosystems. Here, we compare methods of TP estimations in the Lake Huron-Erie corridor (HEC), a system where high diversity of prey items has contributed to the concern that foraging ecology of piscivorous fish species is poorly understood. Using δ15N and δ13C, we quantified TP of longnose gar (Lepisosteus osseus), largemouth bass (Micropterus salmoides), and northern pike (Esox lucius) to assess the efficacy of a scaled DTDF compared to traditional DTDF isotope methods and stomach content analysis (SCA). The scaled DTDF method produced TP estimates that were at times consistent with SCA and were generally higher and with a greater range among individuals than non-scaled DTDFs. The scaled method was not sensitive to baseline choice nor influenced by incorporating carbon source in the model. Greater variability of TP estimates using a scaled DTDF suggests more complex trophic structuring in the upper trophic level guild of the HEC. These results, particularly the lack of baseline sensitivity, provide support for using the scaled DTDF in freshwater food web characterization.

Diet-tissue discrimination factors (δ15 N and δ13 C values) for blood components in Magellanic (Spheniscus magellanicus) and southern rockhopper (Eudyptes chrysocome) penguins.

Analysis of the stable isotope ratios of carbon and nitrogen (δ13 C and δ15 N values) is increasingly being used to gain insight into predator trophic ecology, which requires accurate diet-tissue discrimination factors (DTDFs), or the isotopic difference between prey and predator. Accurate DTDFs must be calculated from predators consuming an isotopically constant diet over time in controlled feeding experiments, but these studies have received little attention to date, especially among seabird species. In this study, aquarium-housed Magellanic (Spheniscus magellanicus) and southern rockhopper (Eudyptes chrysocome) penguins were fed a single-prey source diet (capelin Mallotus villosus) for eight weeks. Stable isotope ratios (δ13 C and δ15 N values) of penguin blood (cellular component and plasma) and capelin were measured using mass spectrometry and then used to calculate DTDFs for both components of penguin blood by comparison with prey values. The DTDFs for plasma were -0.63 ± 0.49 (mean ± SD) and -0.27 ± 0.22 for δ13 C values, and 2.60 ± 0.50 and 2.78 ± 0.22 for δ15 N values for Magellanic and southern rockhopper penguins, respectively, while the DTDFs for the cellular component were 1.22 ± 0.03 and 1.26 ± 0.03 for δ13 C values, and 2.54 ± 0.07 and 2.43 ± 0.17 for δ15 N values. We compare our DTDFs with published values from blood components of penguins and discuss the effects that lipid extraction, sample storage, and diet have on the DTDFs of penguin blood components. This study provides accurate DTDFs of blood components for two seabird species of conservation concern, and is one of the first to provide plasma DTDFs for penguins, which are underrepresented in the seabird literature.

Stable Isotope Analyses of Multiple Tissues of Great Shearwaters (Ardenna Gravis) Reveals Long-Term Dietary Stability, Short-Term Changes in Diet, and Can be Used as a Tool to Monitor Food Webs

The great shearwater (Ardenna gravis) is a common pelagic bird with a distribution that spans almost the entire Atlantic basin, which in conjunction with its relatively high abundance, makes great shearwaters an effective bio indicator. We compared δ13C and δ15N values from the feathers, red blood cells (RBCs), and plasma of great shearwaters collected in 2014 and 2015 from the waters off Massachusetts and Cape Cod. The δ13C and δ15N values of RBCs were quite constant between sampling periods and years, suggesting a generally stable food web over that time period. However, the δ13C of plasma indicates a small seasonal change in diet between July and September for both years, with plasma δ15N values suggesting a slight increase in trophic level late in summer. Comparison of the δ15N of RBCs and plasma indicates that great shearwaters experienced a diet shift during the first few weeks of summer 2014, but not in 2015. Comparisons with other studies suggest that these shearwaters feed at a lower trophic level than great shearwaters sampled in the Bay of Fundy and that there is a decrease in δ13C with increasing latitude, which could indicate a more pelagic diet in northern waters. Stable isotope analysis of the sixth primary feathers provided evidence that these feathers are molted in the Northern Hemisphere and that the diet of great shearwaters shortly after arrival was different in 2014 and 2015. This study demonstrates that within species comparisons of tissue isotopic signatures over time and comparisons of isotopic signatures of tissues with different turnover rates, can detect changes in diet and be used as a tool to monitor for changes in marine food webs over time and space. The relevant signals remain informative even in the absence of species-specific data on tissue-diet discrimination factors, tissue turnover rates, or knowledge of dietary components and their stable isotopic signatures, suggesting dietary changes indicative of a corresponding change in the food web.

Isotopic turnover rates and diet-tissue discrimination depend on feeding habits of freshwater snails.

Estimates of animal diets and trophic structure using stable isotope analysis are strongly affected by diet-tissue discrimination and tissue turnover rates, yet these factors are often unknown for consumers because they must be measured using controlled-feeding studies. Furthermore, these parameters may be influenced by diet quality, growth, and other factors. We measured the effect of dietary protein content on diet-tissue discrimination and tissue turnover in three freshwater snail species. We fed lettuce to individually housed snails (n = 450 per species) for ten weeks, then half were switched to a high-protein diet. Isotopic values of muscle and gonad tissue were assessed at 48 and 80 days post-diet change. Snail discrimination factors varied by diet (low-protein > high-protein) and usually differed among species for both N and C, although species had similar carbon discrimination when fed the low-protein diet. Carbon turnover rates were similar among species for a given tissue type, but nitrogen turnover varied more among species. In addition, diet affected growth of species differently; some species grew larger on high-protein (H. trivolvis) while others grew larger on low-protein diet (Lymnaea spp.). These differences among species in growth influenced turnover rates, which were faster in the species with the highest growth rate following the diet switch from low to high-protein. Thus, growth is one of the main processes that affects tissue turnover, but growth and feeding preference did not affect diet-tissue discrimination, which was greater on low-protein than high-protein diets for all species regardless of growth performance. These results suggest that diet might influence two key parameters of stable isotope analysis differently.

Use of epidermal mucus in elasmobranch stable isotope studies: a pilot study using the giant manta ray (Manta birostris)

Stable isotope analysis of tissues with different turnover rates allows for a broader temporal view of a species’ feeding habits. Epidermal mucus is a rapid to medium turnover ‘tissue’ in teleost fish, but its use in elasmobranch dietary studies is unknown. In the present study, we conducted stable isotope analysis on mucus and muscle from the giant manta ray Manta birostris. Mucus δ13C values were depleted and closer to surface zooplankton δ13C values compared with muscle, whereas there was no significant difference in δ15N values between these two tissue types. Using diet tissue discrimination factors from the literature, there was no significant difference in the expected prey profile between muscle and epidermal mucus. However, a broader range in δ13C values of expected prey was shown for mucus compared with muscle. The results suggest that if M. birostris mucus is indicative of recent dietary intake, resource use during aggregative behaviour off Ecuador is broader, but with no obvious resource switching. The present study is the first example of using bulk stable isotope analysis to evaluate mucus to investigate feeding ecology in elasmobranchs. However, the time course for the change in mucus isotope signature still needs to be determined through controlled feeding studies in an aquarium setting.

Tissue-specific turnover and diet-tissue discrimination factors of carbon and nitrogen isotopes of a common forage fish held at two temperatures.

The application of stable isotopes to foraging ecology is dependent on understanding life-history and environmental factors unrelated to diet that may influence isotopic composition. Diet-tissue discrimination factors (DTDFs) and turnover rates will increase the accuracy of isotope-based studies. Furthermore, little consideration has been given to the effects of temperature or life-history stage on isotopic ratios despite the prevalence of variation in temperature and growth rates throughout life. We measured δ13 C and δ15 N values with an elemental analyzer coupled to a continuous flow isotope ratio mass spectrometer. These values were used to estimate turnover and DTDFs for Emerald Shiners (Notropis atherinoides), a common North American freshwater forage fish. Fish were assigned to a temperature treatment, either 10°C (Low) or 20°C (High), and provided one of three diets (commercial pellet, Artemia salina, or Hemimysis anomala). At regular intervals fish were sampled and the isotopic compositions of whole body and liver tissues were determined. Tissue turnover rates for fish fed Artemia were faster for liver than for whole body, but were also influenced by temperature. Turnover occurred faster at higher temperatures for body and liver δ15 N values, but not for δ13 C values. The pellet and Hemimysis treatments were in isotopic equilibrium from the start of the experiment and estimated DTDFs based on these treatments were lower than assumed for Δ15 N (+0.6 to 2.7‰) and variable, but within expected ranges for Δ13 C (-1.9 to +1.5‰). The results for Emerald Shiners differed from commonly made assumptions for applying stable isotopes to ecological questions, possibly related to a bias in the use of juveniles in studies of turnover and DTDFs and assumptions regarding thermal-independence of isotopic relationships. The species-specific DTDF and tissue turnover estimates provided here will inform interpretations of stable isotope data for smaller fish species and improve food-web studies.

Isotope (δ13C, δ15N, δ2H) diet–tissue discrimination in African grey parrot Psittacus erithacus: implications for forensic studies

ABSTRACTDiet–tissue isotopic relationships established under controlled conditions are informative for determining the dietary sources and geographic provenance of organisms. We analysed δ13C, δ15N, and non-exchangeable δ2H values of captive African grey parrot Psittacus erithacus feathers grown on a fixed mixed-diet and borehole water. Diet–feather Δ13C and Δ15N discrimination values were +3.8 ± 0.3 ‰ and +6.3 ± 0.7 ‰ respectively; significantly greater than expected. Non-exchangeable δ2H feather values (−62.4 ± 6.4 ‰) were more negative than water (−26.1 ± 2.5 ‰) offered during feather growth. There was no positive relationship between the δ13C and δ15N values of the samples along each feather with the associated samples of food offered, or the feather non-exchangeable hydrogen isotope values with δ2H values of water, emphasising the complex processes involved in carbohydrate, protein, and income water routing to feather growth. Understanding the isotopic relationship between diet and feathers may provide greater clarity in the use of stable isotopes in feathers as a tool in determining origins of captive and wild-caught African grey parrots, a species that is widespread in aviculture and faces significant threats to wild populations. We suggest that these isotopic results, determined even in controlled laboratory conditions, be used with caution.