Variation in stable‐isotope ratios between fin and muscle tissues can alter assessment of resource use in tropical river fishes

Carbon and nitrogen stable‐isotope ratios were compared of fin and muscle tissue from 15 fish species collected from seven headwater rivers in eastern and western Thailand. In addition, two‐source stable‐isotope mixing models were used to derive estimates of each fish's reliance on allochthonous and autochthonous energy based on fin and muscle tissues. Across the dataset, fish fin was enriched in ¹³C relative to muscle by c. 1·5‰. Variation in δ¹⁵N between tissues was below statistically significant levels. Estimates of autochthonous resource use calculated from fin tissue were on average 15% greater than those calculated from muscle. Linear mixed‐effects models indicated that inter‐tissue variation in estimates of resource use was predominantly related to inter‐tissue variation in δ¹³C. Fish fin is a credible and desirable alternative to tissues such as muscle or liver which require destructive sampling of fishes. Care must be taken, however, when estimating resource use or interpreting previous estimates of resource use derived from different tissues.     

同位素富集-稀释法研究食性转变对鱼类不同组织N同位素转化率的影响

稳定同位素技术广泛地用于描绘生态系统中食物网的食物来源和营养级关系,但是消费者不同组织转化率的研究相对较少。通过锦鲤摄食人工添加15N蓝藻的食性转化实验,研究不同组织N同位素转化率的差异,探讨组织生长和代谢对同位素转化的相对贡献,为不同时间尺度的稳定同位素研究取样奠定基础。结果表明,通过42d的加富蓝藻饲喂,各组织的N稳定同位素发生显著变化。肝的δ15N为(19.3±1.4)‰,显著高于其它组织,其次为鱼鳍((15.6±1.0)‰)和血液((12.6±0.4)‰),肌肉的δ15N‰最低,为(9.9±0.7)‰。在随后的同位素稀释实验中,锦鲤的体重增加,相对生长速率为0.011d-1,鳍肉的转化率最快,达到11.4%/d,半衰期仅为6.1d,其次是血液和肝,肌肉的转化率最低,仅有3.8%/d,半衰期最长,为18.4d。代谢衰减指数c和-1不存在显著差异,表明锦鲤各组织的N同位素转化主要由组织生长引起。结论显示,同位素富集-稀释法可以有效评价鱼类食性转变对不同组织同位素转化的差异,鳍肉和血液同位素分析可以作为锦鲤食性转变快速追踪的手段。     

Effects of elemental composition on the incorporation of dietary nitrogen and carbon isotopic signatures in an omnivorous songbird

The use of stable isotopes to infer diet requires quantifying the relationship between diet and tissues and, in particular, knowing of how quickly isotopes turnover in different tissues and how isotopic concentrations of different food components change (discriminate) when incorporated into consumer tissues. We used feeding trials with wild-caught yellow-rumped warblers (Dendroica coronata) to determine delta15N and delta13C turnover rates for blood, delta15N and delta13C diet-tissue discrimination factors, and diet-tissue relationships for blood and feathers. After 3 weeks on a common diet, 36 warblers were assigned to one of four diets differing in the relative proportion of fruit and insects. Plasma half-life estimates ranged from 0.4 to 0.7 days for delta13C and from 0.5 to 1.7 days for delta15N . Half-life did not differ among diets. Whole blood half-life for delta13C ranged from 3.9 to 6.1 days. Yellow-rumped warbler tissues were enriched relative to diet by 1.7-3.6% for nitrogen isotopes and by -1.2 to 4.3% for carbon isotopes, depending on tissue and diet. Consistent with previous studies, feathers were the most enriched and whole blood and plasma were the least enriched or, in the case of carbon, slightly depleted relative to diet. In general, tissues were more enriched relative to diet for birds on diets with high percentages of insects. For all tissues, carbon and nitrogen isotope discrimination factors increased with carbon and nitrogen concentrations of diets. The isotopic signature of plasma increased linearly with the sum of the isotopic signature of the diet and the discrimination factor. Because the isotopic signature of tissues depends on both elemental concentration and isotopic signature of the diet, attempts to reconstruct diet from stable isotope signatures require use of mixing models that incorporate elemental concentration.     

Reply to Hussey et al.: The requirement for accurate diet-tissue discrimination factors for interpreting stable isotopes in sharks

Carbon and nitrogen stable isotope analyses have improved our understanding of food webs and movement patterns of aquatic organisms. These techniques have recently been applied to diet studies of elasmobranch fishes, but isotope turnover rates and isotope diet–tissue discrimination are still poorly understood for this group. We performed a diet switch experiment on captive sandbar sharks (Carcharhinus plumbeus) as a model shark species to determine tissue turnover rates for liver, whole blood, and white muscle. In a second experiment, we subjected captive coastal skates (Leucoraja spp.) to serial salinity reductions to measure possible impacts of tissue urea content on nitrogen stable isotope values. We extracted urea from spiny dogfish (Squalus acanthias) white muscle to test for effects on nitrogen stable isotopes. Isotope turnover was slow for shark tissues and similar to previously published estimates for stingrays and teleost fishes with low growth rates. Muscle isotope data would likely fail to capture seasonal migrations or diet switches in sharks, while liver and whole blood would more closely reflect shorter term movement or shifts in diet. Nitrogen stable isotope values of skate blood and skate and dogfish white muscle were not affected by tissue urea content, suggesting that available diet–tissue discrimination estimates for teleost fishes with similar physiologies would provide accurate estimates for elasmobranchs.     

From food to offspring down: tissue-specific discrimination and turn-over of stable isotopes in herbivorous waterbirds and other avian foraging guilds

Isotopic discrimination and turn-over are fundamental to the application of stable isotope ecology in animals. However, detailed information for specific tissues and species are widely lacking, notably for herbivorous species. We provide details on tissue-specific carbon and nitrogen discrimination and turn-over times from food to blood, feathers, claws, egg tissues and offspring down feathers in four species of herbivorous waterbirds. Source-to-tissue discrimination factors for carbon (δ¹³C) and nitrogen stable isotope ratios (δ¹⁵N) showed little variation across species but varied between tissues. Apparent discrimination factors ranged between −0.5 to 2.5‰ for δ¹³C and 2.8 to 5.2‰ for δ¹⁵N, and were more similar between blood components than between keratinous tissues or egg tissue. Comparing these results with published data from other species we found no effect of foraging guild on discrimination factors for carbon but a significant foraging-guild effect for nitrogen discrimination factors.Turn-over of δ¹³C in tissues was most rapid in blood plasma, with a half-life of 4.3 d, whereas δ¹³C in blood cells had a half-life of approximately 32 d. Turn-over times for albumen and yolk in laying females were similar to those of blood plasma, at 3.2 and 6.0 d respectively. Within yolk, we found decreasing half-life times of δ¹³C from inner yolk (13.3 d) to outer yolk (3.1 d), related to the temporal pattern of tissue formation.We found similarities in tissue-specific turn-over times across all avian species studied to date. Yet, while generalities regarding discrimination factors and tissue turn-over times can be made, a large amount of variation remains unexplained.     

Tissue Turnover Rates and Isotopic Trophic Discrimination Factors in the Endothermic Teleost,Pacific Bluefin Tuna (Thunnus orientalis)

Stable isotope analysis (SIA) of highly migratory marine pelagic animals can improve understanding of their migratory patterns and trophic ecology. However, accurate interpretation of isotopic analyses relies on knowledge of isotope turnover rates and tissue-diet isotope discrimination factors. Laboratory-derived turnover rates and discrimination factors have been difficult to obtain due to the challenges of maintaining these species in captivity. We conducted a study to determine tissue- (white muscle and liver) and isotope- (nitrogen and carbon) specific turnover rates and trophic discrimination factors (TDFs) using archived tissues from captive Pacific bluefin tuna (PBFT), Thunnus orientalis, 1–2914 days after a diet shift in captivity. Half-life values for ¹⁵N turnover in white muscle and liver were 167 and 86 days, and for ¹³C were 255 and 162 days, respectively. TDFs for white muscle and liver were 1.9 and 1.1‰ for δ ¹⁵N and 1.8 and 1.2‰ for δ ¹³C, respectively. Our results demonstrate that turnover of ¹⁵N and ¹³C in bluefin tuna tissues is well described by a single compartment first-order kinetics model. We report variability in turnover rates between tissue types and their isotope dynamics, and hypothesize that metabolic processes play a large role in turnover of nitrogen and carbon in PBFT white muscle and liver tissues. ¹⁵N in white muscle tissue showed the most predictable change with diet over time, suggesting that white muscle δ ¹⁵N data may provide the most reliable inferences for diet and migration studies using stable isotopes in wild fish. These results allow more accurate interpretation of field data and dramatically improve our ability to use stable isotope data from wild tunas to better understand their migration patterns and trophic ecology.     

Turnover rates of nitrogen stable isotopes in the salt marsh mummichog, Fundulus heteroclitus, following a laboratory diet switch

Nitrogen stable isotopes are frequently used in ecological studies to estimate trophic position and determine movement patterns. Knowledge of tissue-specific turnover and nitrogen discrimination for the study organisms is important for accurate interpretation of isotopic data. We measured δ¹⁵ N turnover in liver and muscle tissue in juvenile mummichogs, Fundulus heteroclitus, following a laboratory diet switch. Liver tissue turned over significantly faster than muscle tissue suggesting the potential for a multiple tissue stable isotope approach to study movement and trophic position over different time scales; metabolism contributed significantly to isotopic turnover for both liver and muscle. Nitrogen diet-tissue discrimination was estimated at between 0.0 and 1.2‰ for liver and –1.0 and 0.2‰ for muscle. This is the first experiment to demonstrate a significant variation in δ¹⁵ N turnover between liver and muscle tissues in a fish species.     

Tissue-specific turnover rates of the nitrogen stable isotope as functions of time and growth in a cyprinid fish

Ecological applications of stable isotope data require knowledge on the isotopic turnover rate of tissues, usually described as the isotopic half-life in days (T _0.5) or the change in mass (G _0.5). Ecological studies increasingly analyse tissues collected non-destructively, such as fish fin and scales, but there is limited knowledge on their turnover rates. Determining turnover rates in situ is challenging, with ex situ approaches preferred. Correspondingly, T _0.5 and G _0.5 of the nitrogen stable isotope (δ¹⁵N) were determined for juvenile barbel Barbus barbus (5.5 ± 0.6 g starting weight) using a diet-switch experiment. δ¹⁵N data from muscle, fin and scales were taken during a 125 day post diet-switch period. Whilst isotopic equilibrium was not reached in the 125 days, the δ¹⁵N values did approach those of the new diet. The fastest turnover rates were in more metabolically active tissues, from muscle (highest) to scales (lowest). Turnover rates were relatively slow; T _0.5 was 84 (muscle) to 145 (scale) days; G _0.5 was 1.39 × body mass (muscle) to 2.0 × body mass (scales), with this potentially relating to the slow growth of the experimental fish. These turnover estimates across the different tissues emphasise the importance of estimating half-lives for focal taxa at species and tissue levels for ecological studies.     

Non-lethal dorsal fin sampling for stable isotope analysis in seahorses

Sampling collection for stable isotope analysis has traditionally involved the sacrifice of the animal. Seahorses (Hippocampus spp.) are listed as threatened by the Convention on International Trade in Endangered Species (http://www.cites.org) and consequently lethal sampling is undesirable. We evaluated the adequacy of dorsal fin tissue of adult seahorses Hippocampus guttulatus for stable isotope analysis as an alternative to lethal tissue sampling. Three seahorse tissues (dorsal fin, muscle, and liver) were analyzed for comparisons of δ¹⁵N and δ¹³C values. Similarities found between δ¹⁵N and δ¹³C values in dorsal fin and muscle tissue of H. guttulatus suggest that both tissues are adequate for stable isotope analysis to understand feeding ecology of seahorses. However, considering the threatened status of the species, dorsal fin tissue would be recommended in adult seahorses as a non-lethal sampling. The effect of lipid extraction on carbon and nitrogen stable isotope values was also evaluated in each seahorse tissues. Significant effects of lipids extraction did only occur for δ¹³C values in muscle and liver. It was found that lipid removal was not necessary to perform SIA in dorsal fin tissues. Due to the limited availability of fin tissue obtained from fin-clipping in seahorses, the relationship between the mass/surface of dorsal fin clip and stable isotope values was analyzed. δ¹⁵N and δ¹³C values in fin samples were found to be independent of the size of fin analyzed. According to our study, the use of fin-clipping sampling, with a minimum surface analyzed of 12.74?mm², was found to be an adequate method for SIA in seahorses.     

Variation in carbon and nitrogen isotopic ratios of fin and muscle tissues of Longnose Gar (Lepisosteus osseus) and Smallmouth Buffalo (Ictiobus bubalus)

Fin clips have been proposed as a non-lethal and non-invasive alternative to dorsal muscle samples in stable isotope analysis. However, potential differences in elemental composition and turnover rates can bias inferences when different tissues are combined. Here, we tested the average difference and correlation of the isotopic signature of δ¹³C and δ¹⁵N between muscle and fin samples in two arge-bodied fishes: Longnose Gar (Lepisosteus osseus) and Smallmouth Buffalo (Ictiobus bubalus). We found that δ¹⁵N signatures of muscle and fin tissues were strongly and positively correlated for both species, although the muscle tissue for Smallmouth Buffalo was slightly more enriched in δ15N. δ¹³C signatures of both tissues were significatively different and not strongly correlated for Longnose Gar, but similar and strongly correlated for Smallmouth Buffalo. Our results suggest that fin and muscle tissue can be combined for analyses of δ¹⁵N, but correction for higher enrichment of muscle tissue may be necessary for Smallmouth Buffalo. Conversely, combining fin and muscle tissue for analysis of δ¹³C requires more caution due to their weaker correlation and dependence of species identity.     

Estimating the timing of diet shifts using stable isotopes

Stable isotope analysis has become an important tool in studies of trophic food webs and animal feeding patterns. When animals undergo rapid dietary shifts due to migration, metamorphosis, or other reasons, the isotopic composition of their tissues begins changing to reflect that of their diet. This can occur both as a result of growth and metabolic turnover of existing tissue. Tissues vary in their rate of isotopic change, with high turnover tissues such as liver changing rapidly, while relatively low turnover tissues such as bone change more slowly. A model is outlined that uses the varying isotopic changes in multiple tissues as a chemical clock to estimate the time elapsed since a diet shift, and the magnitude of the isotopic shift in the tissues at the new equilibrium. This model was tested using published results from controlled feeding experiments on a bird and a mammal. For the model to be effective, the tissues utilized must be sufficiently different in their turnover rates. The model did a reasonable job of estimating elapsed time and equilibrial isotopic changes, except when the time since the diet shift was less than a small fraction of the half-life of the slowest turnover tissue or greater than 5–10 half-lives of the slowest turnover tissue. Sensitivity analyses independently corroborated that model estimates became unstable at extremely short and long sample times due to the effect of random measurement error. Subject to some limitations, the model may be useful for studying the movement and behavior of animals changing isotopic environments, such as anadromous fish, migratory birds, animals undergoing metamorphosis, or animals changing diets because of shifts in food abundance or competitive interactions.     

Fin tissues as surrogates of white muscle when assessing carbon and nitrogen stable isotope levels for Arctic and brook char

Arctic char (Salvelinus alpinus) are a fish species ubiquitous to the fresh waters of Arctic region and brook char (Salvelinus fontinalis) are similarly common across the sub-Arctic region of eastern Canada. Populations can be small in numbers, especially farther north thus it is important to develop non-lethal methods of sampling these fish to minimize the invasiveness and impact of scientific research. We examined the stable isotopes of nitrogen and carbon in white muscle, caudal fin, and adipose fin tissues of Arctic char and brook char (S. fontinalis) from northern Quebec and Labrador, Canada. Our results revealed several broad conclusions. First, differences among muscle, caudal fin, and adipose fin tissues were ~1?‰ for freshwater Arctic and brook char. Second, the two species within the same drainage had similar stable isotope levels and tissue differences. Third, anadromous Arctic char show similar, non-significant differences among these tissues for δ¹⁵N, but muscle δ¹³C was highly enriched. Fourth, the stable isotope levels and tissue differences were the same for anadromous Arctic char from two watersheds where char use distinctly different ocean environments. Overall, it appears that caudal fin tissue in particular is a useful surrogate for white muscle δ¹³C and δ¹⁵N levels for Arctic and brook char in this region and thus, a non-lethal collection of a small sample of caudal fin tissue will provide an accurate measure of white muscle isotope levels.     

Intrapopulation variability shaping isotope discrimination and turnover: experimental evidence in arctic foxes

Background

Tissue-specific stable isotope signatures can provide insights into the trophic ecology of consumers and their roles in food webs. Two parameters are central for making valid inferences based on stable isotopes, isotopic discrimination (difference in isotopic ratio between consumer and its diet) and turnover time (renewal process of molecules in a given tissue usually measured when half of the tissue composition has changed). We investigated simultaneously the effects of age, sex, and diet types on the variation of discrimination and half-life in nitrogen and carbon stable isotopes (δ¹⁵N and δ¹³C, respectively) in five tissues (blood cells, plasma, muscle, liver, nail, and hair) of a top predator, the arctic fox Vulpes lagopus.

Methodology/Principal Findings

We fed 40 farmed foxes (equal numbers of adults and yearlings of both sexes) with diet capturing the range of resources used by their wild counterparts. We found that, for a single species, six tissues, and three diet types, the range of discrimination values can be almost as large as what is known at the scale of the whole mammalian or avian class. Discrimination varied depending on sex, age, tissue, and diet types, ranging from 0.3‰ to 5.3‰ (mean  = 2.6‰) for δ¹⁵N and from 0.2‰ to 2.9‰ (mean  = 0.9‰) for δ¹³C. We also found an impact of population structure on δ¹⁵N half-life in blood cells. Varying across individuals, δ¹⁵N half-life in plasma (6 to 10 days) was also shorter than for δ¹³C (14 to 22 days), though δ¹⁵N and δ¹³C half-lives are usually considered as equal.

Conclusion/Significance

Overall, our multi-factorial experiment revealed that at least six levels of isotopic variations could co-occur in the same population. Our experimental analysis provides a framework for quantifying multiple sources of variation in isotopic discrimination and half-life that needs to be taken into account when designing and analysing ecological field studies.     

Non-lethal sampling for stable isotope analysis of pike Esox lucius: how mucus,scale and fin tissue compare to muscle

Stable isotope analysis (SIA) was used to examine the isotopic relationships between dorsal muscle and fin, scale and epidermal mucus in pike Esox lucius. δ¹³C and δ¹⁵N varied predictably within each tissue pairing, with conversion factors calculated for the surrogate tissues, enabling their application to the non-lethal sampling of E. lucius for SIA.     

Individual variability in stable isotope turnover rates of epidermal mucus according to body size in an omnivorous fish

Epidermal mucus (‘mucus’) is increasingly applied to fish ecological studies based on stable isotope analysis (SIA) due to its non-invasive collection. However, knowledge on mucus SI turnover rates of individual fish remains limited, including uncertainty over how they are influenced by fish body sizes. Here, a diet switch experiment predicted mucus SI turnover rates (δ¹³C and δ¹⁵N) as a function of time using samples taken over 200 days from 10 individually tagged common carp Cyprinus carpio covering two size groups. Non-linear mixed effects models revealed rapid turnover of both δ¹³C and δ¹⁵N (T₅₀: 2–5 days; T₉₅: 9–22 days); δ¹⁵N turnover rates were slower for the larger cohort, while δ¹³C turnover rates were independent of body size. Within size groups, turnover rates were not expected to vary between individuals. These experimental results suggest that due to these fast turnover rates, epidermal mucus can provide insights into the diets of fish over very short timeframes, although for δ¹⁵N the body size of the fish needs consideration.

     

Dual‐tracer‐based isotope turnover rates in a highly invasive mysid Limnomysis benedeni from Lake Constance

Understanding the ecological patterns of invasive species and their habitats require an understanding of the species’ foraging ecology. Stable carbon (δ¹³C) and nitrogen (δ¹⁵N) isotope values provide useful information into the study of animal ecology and evolution, since the isotope ratios of consumers reflect consumer's dietary patterns. Nevertheless, the lack of species‐ and element‐specific laboratory‐derived turnover rates could limit their application. Using a laboratory‐based dual stable isotope tracer approach (Na¹⁵NO₃ and NaH¹³CO₃), we evaluated the δ¹⁵N and δ¹³C isotope turnover rates in full‐grown adult invasive Limnomysis benedeni from Lake Constance. We provide δ¹⁵N and δ¹³C turnover rates based on nonlinear least‐squares regression and posterior linear regression models. Model precisions and fit were evaluated using Akaike's information criterion. Within a couple of days, the δ¹⁵N and δ¹³C of mysids began to change. Nevertheless, after about 14 days, L. benedeni did not reach equilibrium with their new isotope values. Since the experiment was conducted on adult subjects, it is evident that turnover was mainly influenced by metabolism (in contrast to growth). Unlike traditional dietary shifts, our laboratory‐based dual stable isotope tracer approach does not shift the experimental organisms into a new diet and avoids dietary effects on isotope values. Results confirm the application of isotopic tracers to label mysid subpopulations and could be used to reflect assimilation and turnover from the labeled dietary sources. Field‐based stable isotope studies often use isotopic mixing models commonly assuming diet‐tissue steady state. Unfortunately, in cases where the isotopic composition of the animal is not in equilibrium with its diet, this can lead to highly misleading conclusions. Thus, our laboratory‐based isotopic incorporation rates assist interpretation of the isotopic values from the field and provide a foundation for future research into using isotopic tracers to investigate invasion ecology.     

Consumer‐resource stoichiometry as a predictor of trophic discrimination (Δ13C, Δ15N) in aquatic invertebrates

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Temperature and diet affect carbon and nitrogen isotopes of fish muscle: can amino acid nitrogen isotopes explain effects?

Stable isotope ratios of carbon (δ¹³C) and nitrogen (δ¹⁵N) are widely used in food-web studies to determine trophic positioning and diet sources. However in order to accurately interpret stable isotope data the effects of environmental variability and dietary composition on isotopic discrimination factors and tissue turnover rates must be validated. We tested the effects of temperature and diet on tissue turnover rates and discrimination of carbon and nitrogen isotopes in an omnivorous fish, black bream (Acanthopagrus butcheri). Fish were raised at 16 °C or 23 °C and fed either a fish-meal or vegetable feed to determine turnover rates in fish muscle tissue up to 42 days after exposure to experimental treatments. Temperature and diet affected bulk tissue δ¹⁵N turnover and discrimination factors, with increased turnover and smaller discrimination factors at warmer temperatures. Fish reared on the vegetable feed showed greater bulk tissue δ¹⁵N changes and larger discrimination factors than those reared on a fish-meal feed. Temperature and diet affected bulk tissue δ¹³C values, however the direction of effects among treatments changed. Analyses of δ¹⁵N values of individual amino acids found few significant changes over time or treatment effects, as there was large variation at the individual fish level. However glutamic acid, aspartic acid and leucine changed most over the experiment and results mirrored those of treatment effects in bulk δ¹⁵N tissue values. The results demonstrate that trophic discrimination for δ¹⁵N and δ¹³C can be significantly different than those typically used in food-web analyses, and effects of diet composition and temperature can be significant. Precision of compound-specific isotope analyses (0.9‰) was larger than our effect size for bulk δ¹⁵N diet effects (0.7‰), therefore future experimental work in this area will need to establish a large effect size in order to detect significant differences. Our results also suggest that compound-specific amino acid δ¹⁵N may be useful for determining essential and non-essential amino acids for different animals.     

Application of non‐lethal stable isotope analysis to assess feeding patterns of juvenile pallid sturgeon Scaphirhynchus albus: a comparison of tissue types and sample preservation methods

Traditional techniques for stable isotope analysis (SIA) generally require sacrificing animals to collect tissue samples; this can be problematic when studying diets of endangered species such as the pallid sturgeon Scaphirhynchus albus. Our objectives were to (i) determine if pectoral fin tissue (non‐lethal) could be a substitute for muscle tissue (lethal) in SIA of juvenile pallid sturgeon, and (ii) evaluate the influence of preservation techniques on stable isotope values. In the laboratory, individual juvenile pallid sturgeon were held for up to 186 day and fed chironomids, fish, or a commercially available pellet diet. Significant, positive relationships (r² ≥ 0.8) were observed between fin and muscle tissues for both δ¹⁵N and δ¹³C; in all samples isotopes were enriched in fins compared to muscle tissue. Chironomid and fish based diets of juvenile pallid sturgeon were distinguishable for fast growing fish (0.3 mm day^?1) using stable δ¹⁵N and δ¹³C isotopes. Frozen and preserved fin tissue δ¹⁵N isotopes were strongly related (r² = 0.89) but δ¹³C isotopes were weakly related (r² = 0.16). Therefore, freezing is recommended for preservation of fin clips to avoid the confounding effect of enrichment by ethanol. This study demonstrates the utility of a non‐lethal technique to assess time integrated food habits of juvenile pallid sturgeon and should be applicable to other threatened or endangered species.     

Fractionation and turnover of stable carbon isotopes in animal tissues: Implications for δ13C analysis of diet

The use of stable carbon isotopes as a means of studying energy flow is increasing in ecology and paleoecology. However, secondary fractionation and turnover of stable isotopes in animals are poorly understood processes. This study shows that tissues of the gerbil (Meriones unguienlatus) have different δ¹³C values when equilibrated on corn (C₄) or wheat (C₃) diets with constant ¹³C/¹²C contents. Lipids were depleted 3.0‰ and hair was enriched 1.0‰ relative to the C₄ diet. Tissue δ¹³C values were ranked hair>brain>muscle>liver>fat. After changing the gerbils to a wheat (C₃) diet, isotope ratios of the tissues shifted in the direction of the δ¹³C value of the new diet. The rate at which carbon derived from the corn diet was replaced by carbon derived from the wheat diet was adequately described by a negative exponential decay model for all tissues examined. More metabolically active tissues such as liver and fat had more rapid turnover rates than less metabolically active tissues such as hair. The half-life for carbon ranged from 6.4 days in liver to 47.5 days in hair. The results of this study have important implications for the use of δ¹³C values as indicators of animal diet. Both fractionation and turnover of stable carbon isotopes in animal tissues may obscure the relative contributions of isotopically distinct dietary components (such as C₃ vs. C₄, or marine vs. terrestrial) if an animal's diet varies through time. These complications deserve attention in any study using stable isotope ratios of animal tissue as dietary indicators and might be minimized by analysis of several tissues or products covering a range of turnover times.