The stable isotope ecology of terrestrial plant succession

We review the relevance and use of stable isotopes for the study of plant community succession. Stable isotope measurements provide information on the origin of resources acquired by plants, the processes governing resource uptake and transformation, and the physiological and environmental conditions of plant growth. When combined with measurements of the stable isotope ratio values of soil microbial biomass, soil organic matter and plant litter, isotope measurements of plants can indicate effects of successional changes on ecosystem processes. However, their application to questions of plant succession and ecosystem change is limited by the degree to which the underlying assumptions are met in each study, and complementary measures may be required, depending upon the question of interest. First, we discuss the changes that occur in the stable isotope composition of plants and ecosystems with ontogeny and species replacements, as well as their potential evolutionary significance. Second, we discuss the imprints of plant competition and facilitation on leaf and wood tissue, as well as how stable isotopes can provide novel insights on the mechanisms underlying plant interactions. Finally, we discuss the capacity for stable isotope measurements to serve as a proxy record for past disturbances such as fire, logging and cyclones.     

Significance of diet type and diet quality for ecological diversity of African ungulates

1. We test two nutritional hypotheses for the ecological diversity of ungulates, the browser/grazer (diet type) and diet quality models, among free-ranging herbivores in a South African savanna, the Kruger National Park. Tests are based on assessment of relationships between diet type and diet quality with body mass and hypsodonty, two morphological features that have been associated with both elements. 2. We use stable carbon isotope ratios of faeces to reconstruct diet in terms of proportions of C(3) plants (browse) and C(4) plants (grass) consumed by different species in different seasons. These data are combined with proxies for diet quality (per cent nitrogen, neutral detergent fibre, acid detergent fibre, and acid detergent lignin) from faeces to track changes in diet quality. 3. Two statistical approaches are used in model selection, i.e. tests of significant correlations based on linear regression analyses, and an information-theory approach (Akaike's Information Criterion) providing insight into strength of evidence for models. 4. Results of both methods show that, contrary to many predictions, body mass and diet type are not related, but these data confirm predictions that diet quality decreases with increasing body size, especially during the dry season. Hypsodonty, as expected, varies with diet type, increasing with increased grass intake. 5. These findings support both a diet type and diet quality model, implying some degree of exclusivity. We propose that congruence between models may be achieved through addition of diet quality proxies not included here, because hypsodonty is more likely a reflection of the abrasive properties of consumed foods, i.e. related to food quality, rather than food type. This implies that adaptation to diets of varying quality, through changes in body size and dental features, has been the primary mechanism for diversification in ungulates. 6. Our interpretation contrasts with several recent studies advocating diet type as the primary factor, exemplifying that further reconciliation between the two models is needed. We discuss the implications of this study for future approaches to achieve a more cohesive understanding of the evolutionary outcomes of herbivore nutrition.     

Carbon isotope fractionation between diet and bioapatite in ungulate mammals and implications for ecological and paleoecological studies

 The isotope enrichment ɛ* of ¹³C between tooth enamel of large ruminant mammals and their diet is 14.1 ± 0.5‰. This value was obtained by analyzing both the dental enamel of a variety of wild and captive mammals and the vegetation that comprised their foodstuffs. This isotope enrichment factor applies to a wide variety of ruminant mammals. Non-ruminant ungulates have a similar isotope enrichment, although our data cannot determine if it is significantly different. We also found a ¹³C isotope enrichment ɛ* of 3.1 ± 0.7‰ for horn relative to diet, and 11.1 ± 0.8‰ for enamel relative to horn for ruminant mammals. Tooth enamel is a faithful recorder of diet. Its isotopic composition can be used to track changes in the isotopic composition of the atmosphere, determine the fraction of C₃ or C₄ biomass in diets of modern or fossil mammals, distinguish between mammals using different subpathways of C₄ photosynthesis,and identify those mammals whose diet is derived from closed-canopy habitats. Received: 1 July 1998 / Accepted: 9 February 1999     

Physical and chemical basis of carbon isotope fractionation in plants

Naturally-occurring variations in the abundances of the stable isotopes of carbon and other elements can be used to understand the dynamics of natural processes in chemistry, biochemistry, biology, medicine, ecology and other fields. The use of carbon-13 isotopic abundances as an indicator of photosynthetic function in plants has become common. The purpose of this article is to describe the physical and chemical processes that contribute to the abundances of carbon-13 in plant materials, and to provide a framework for understanding how those processes control the isotopic contents of natural materials.     

Carbon isotope fractionation of methyl bromideduring agricultural soil fumigations

The isotopic composition ofmethyl bromide (CH₃Br) has been suggestedto be a potentially useful tracer forconstraining the global CH₃Br budget. Inorder to determine the carbon isotopiccomposition of CH₃Br emitted from the mostsignificant anthropogenic application(pre-plant fumigation) we directly measured the¹³C of CH₃Br released duringcommercial fumigation. We also measured theisotopic fractionation associated withdegradation in agricultural soil under typicalfield fumigation conditions. The isotopiccomposition of CH₃Br collected in soilseveral hours after injection of the fumigantwas –44.5 and this value increased to –20.7over the following three days. The mean kineticisotope effect (KIE) associated withdegradation of CH₃Br in agricultural soil(12) was smaller than the reported value formethylotrophic bacterial strain IMB-1, isolatedfrom previously fumigated agricultural soil,but was similar to methylotrophic bacterialstrain CC495, isolated from a pristine forestlitter zone. Using this fractionationassociated with the degradation of CH₃Brin agricultural soil and the mean¹³C of the industriallymanufactured CH₃Br (–54.4), we calculatethat the agricultural soil fumigation sourcehas a carbon isotope signature that ranges from–52.8 to –42.0. Roughly 65% ofindustrially manufactured CH₃Br is usedfor field fumigations. The remaining 35% isused for structural and post-harvestfumigations with a minor amount used duringindustrial chemical manufacturing. Assumingthat the structural and post-harvest fumigationsources of CH₃Br are emitted withoutsubstantial fractionation, we calculate thatthe ¹³C of anthropogenicallyemitted CH₃Br ranges from –53.2 to –47.5.     

Experimental study on the effect of diet on fatty acid and stable isotope profiles of the squid Lolliguncula brevis

Fatty acid and stable isotope analyses have previously been used to investigate foraging patterns of fish, birds, marine mammals and most recently cephalopod species. To evaluate the application of these methods for dietary studies in squid, it is important to understand the degree to which fatty acid and stable isotope signatures of prey species are reflected in the squids' tissue. Four groups of Lolliguncula brevis were fed on prey species with distinctly different fatty acid and stable isotope profiles over 30 consecutive days. One group of squid were fed fish for fifteen days, followed by crustaceans for a further fifteen days. A second and third group were fed exclusively on fish or crustaceans for thirty days. And a fourth group was fed on a mixture of fish and crustaceans for thirty days. Analysis of squid tissue showed that, after 10 days of feeding, fatty acid profiles of squid tended to reflect those of their prey. Squid that fed on a single prey type, i.e. fish or crustacean, showed only minor modifications in fatty acid proportions after the initial change and fatty acid profiles were clearly distinguishable between the two feeding groups. Shifts in fatty acid proportions towards respective prey profiles could clearly be observed in squid the diet of which was swapped after 15 days. Clear differences could also be seen in fatty acid profiles of squid feeding on a mixed diet with trends towards either fish or crustacean fatty acid signatures. Stable isotope signatures of squid tissues clearly distinguished between animals feeding on different diets and supported findings from fatty acid analysis, thus indicating both methods to be viable tools in feeding studies on squid species.     

Temporal and intrapopulation variation in prey choice of wintering geese determined by stable isotope analysis

1. Individual variability in prey preferences can have marked effects on many demographic parameters from individual survival and fecundity to the vital rates of entire populations. A population level response is ultimately determined by individual prey choices; however, the effect of individual dietary choice is often overlooked. 2. We determined prey choice by individual consumers, light-bellied Brent geese Branta bernicla, during the overwintering period. Two hundred and eighty-one individuals were sampled at distinct temporal points over two winters. Stable isotopic ratios of carbon and nitrogen for blood cells and blood plasma, from each sampled individual were measured. Isotopic ratios for potential prey items were also measured. 3. Delta15N and delta13C for blood samples were both significantly different between sample months. Generally we found a decrease in both isotopic ratios during the course of the winter. All potential prey items were also isotopically distinct. Multisource mixing models (isosource) were used to determine the range of possible contribution to the diet of individuals. 4. During early winter, diet consisted almost exclusively of sea grass Zostera spp. The level of Zostera spp. in the diet dropped until mid-winter, and was supplemented by the utilization of green algae Ulva lactuca, and Enteromorpha spp., and terrestrial grasses. Terrestrial grass comprised an increasing proportion of the diet in late winter, representing virtually the exclusive food source by April. 5. By examining intrapopulation variability in resource utilization we highlight a number of ecologically important factors not addressed by previous population level studies.     

A stable carbon isotope study of dissolved inorganic carbon cycling in a softwater lake

The dissolved inorganic carbon (DIC) cycle in a softwater lake was studied using natural variations of the stable isotopes of carbon,¹²C and¹³C. During summer stratification there was a progressive decrease in epilimnion DIC concentration with a concomitant increase in ¹³C_DIC), due to preferential uptake of¹²C by phytoplankton and a change in the dominant CO₂ source from inflow andin situ oxidation to invasion from the atmosphere. There was an increase in hypolimnion DIC concentration throughout summer with a concomitant general decrease in ¹³C_DIC from oxidation of the isotopically light particulate organic carbon that sank down through the thermocline from the epilimnion.Mass balance calculations of DI¹²C and DI¹³C in the epilimnion for the summer (June 23–September 25) yield a mean rate of net conversion of DIC to organic carbon (C_org) of 430 ± 150 moles d^-1 (6.5 ± 1.8 m moles m^-2 d^-1. Net CO₂ invasion from the atmosphere was 420 ± 120 moles d^-1 (6.2 ± 1.8 m moles m^-2 d^-1) with an exchange coefficient of 0.6 ± 0.3m d^-1. These results imply that at least for the summer months the phytoplankton obtained about 90% of their carbon from atmosphere CO₂. About 50% of CO₂ invasion and conversion to C_org for the summer occurred during a two week interval in mid-summer.DIC concentration increased in the hypolimnion at a rate of 350 ± 70 moles DIC d^-1 during summer stratification. The amount of DIC added to the hypolimnion was equivalent to 75 ± 20% of net conversion of DIC to C_org in the euphotic zone over spring and summer implying rapid degradation of POC in the hypolimnion. The ¹³C of DIC added to the deep water (-22.) was too heavy to have been derived from oxidation of particulate organic carbon alone. About 20% of the added DIC must have diffused from hypolimnetic sediments where relatively heavy CO₂ (-7) was produced by a combination of POC oxidation and as a by-product of methanogenesis.     

Revolution in food web analysis and trophic ecology: diet analysis by DNA and stable isotope analysis

Characterization of energy flow in ecosystems is one of the primary goals of ecology, and the analysis of trophic interactions and food web dynamics is key to quantifying energy flow. Predator‐prey interactions define the majority of trophic interactions and food web dynamics, and visual analysis of stomach, gut or fecal content composition is the technique traditionally used to quantify predator‐prey interactions. Unfortunately such techniques may be biased and inaccurate due to variation in digestion rates ( Sheppard & Hardwood 2005 ); however, those limitations can be largely overcome with new technology. In the last 20 years, the use of molecular genetic techniques in ecology has exploded ( King et al. 2008 ). The growing availability of molecular genetic methods and data has fostered the use of PCR‐based techniques to accurately distinguish and identify prey items in stomach, gut and fecal samples. In this month’s issue of Molecular Ecology Resources, Corse et al. (2010) describe and apply a new approach to quantifying predator‐prey relationships using an ecosystem‐level genetic characterization of available and consumed prey in European freshwater habitats ( Fig. 1a ). In this issue of Molecular Ecology, Hardy et al. (2010) marry the molecular genetic analysis of prey with a stable isotope (SI) analysis of trophic interactions in an Australian reservoir community ( Fig. 1b ). Both papers demonstrate novel and innovative approaches to an old problem – how do we effectively explore food webs and energy movement in ecosystems?

Figure 1 Open in figure viewer PowerPoint The aquatic habitats used for two studies of diet and trophic interactions that employed molecular genetic and stable isotope analyses. Panel a: Example of Rhone basin habitat (France) where fish diet was determined using PCR to classify prey to a series of ecological clades (photo by Emmanuel Corse). Panel b: A weir pool on the lower Murray River (Australia) where food web and prey use was evaluated using a combination of advanced molecular genetic and stable isotope analyses (photo credit: CSIRO).     

Effect of lipid removal on carbon and nitrogen stable isotope ratios in crustacean tissues

The analysis of tissue's naturally occurring stable carbon and nitrogen isotope ratios is a useful tool to delineate trophic relationships. However, the interpretation of δ¹³C and δ¹⁵N is complicated by the influence of multiple factors such as the tissue-specific lipid content. The aim of this work was to evaluate the effects of lipid extraction on δ¹³C and δ¹⁵N compositions in muscle, hepatopancreas and gonads of a marine decapod crustacean, the spider crab Maja brachydactyla. Samples were analyzed for stable isotopes before and after lipid removal, using a derived Soxhlet extraction method. Differences in δ¹³C and δ¹⁵N were measured among tissues before and after treatment. Lipid extraction of muscle did not have a significant effect on either δ¹³C or δ¹⁵N. By contrast, ecologically significant shifts for both carbon and nitrogen stable isotopes ratios (+ 2.9 ± 0.8‰ for δ¹³C, and + 1.2 ± 0.7‰ for δ¹⁵N) were noticed in the hepatopancreas. In regard to gonads, lipid extraction led to a shift only on δ¹³C (+ 1.3 ± 0.3‰). Finally, the derived Soxhlet extraction method removed the lipid influence for δ¹³C, and had an effect on δ¹⁵N composition for lipid-rich samples. We recommend this treatment for carbon stable isotope studies on decapod crustacean lipid-rich tissues.     

A stable carbon and nitrogen isotope study of lake food webs in Canada's Boreal Plain

1.  Stable carbon and nitrogen isotope and fish stomach content analyses were used to investigate food webs in five relatively undisturbed lakes on the Boreal Plain of Canada. Stable isotope analysis was also used to determine the importance of external and internal carbon sources.
2.  Overlap in the carbon and nitrogen signatures of primary producers made it difficult to determine unambiguously the feeding habits of many invertebrates. However, isotope analysis suggested that external carbon inputs were detectable in the aquatic food chains of the one lake with a short water residence time («1 year). In the other four lakes, with water residence times ≥1 year, autochthonous carbon was the only detectable carbon source in the food webs.
3.  Food webs in these lakes spanned a range of four to five trophic levels. Both invertebrates and fish appeared to eat a variety of food, often feeding at more than one trophic level.
4.  With the exception of one lake (SPH20), top predators in these lakes, northern pike ( Esox lucius ) and fathead minnows ( Pimephales promelas ), occupied similar trophic positions despite large differences in body size and trophic morphology. In SPH20, where there were two additional fish species, pike occupied a higher trophic position. However, all the top predators in each lake appeared to be omnivores and generalists.
5.  The prevalence of omnivory and the apparent generalist feeding habits of fish in these lakes suggest that organisms are flexible in their feeding habits and that these food webs will be resilient to disturbance.     

Assessment of anadromous salmon resources in the diet of the Alexander Archipelago wolf using stable isotope analysis

The Alexander Archipelago wolf (Canis lupus ligoni) is unique to southeast Alaska, occurring on islands south of Frederick Sound and along the mainland between Dixon Entrance and Yakutat Bay. Sitka black-tailed deer (Odocoileus hemionus sitkensis) are an important prey species for wolves across the southern part of the region. Spawning salmon (Onchorynchus sp.) are seasonally available but their presence in wolf diets has not previously been quantified. We examined the range of bone collagen δ¹³C and δ¹⁵N values for wolves throughout southeast (n = 163) and interior (n = 50) Alaska and used a dual-isotope mixing model to determine the relative contribution of salmon-derived marine protein in the diet. Southeast Alaska wolves consumed significantly more salmon (mean ± SE: 18.3 ± 1.2%) than did wolves from interior Alaska (9.1 ± 0.6%, P<0.001). Wolves on the southeast Alaska mainland appeared to have higher marine isotopic signatures than island wolves, although this difference was not significant. Variation among individual wolf diets was higher for southeast than for interior Alaska wolves, and variation was highest in coastal mainland wolf diets (P<0.001). Marine resources may augment the diet of southeast Alaska wolves during seasonal or annual fluctuations in the availability of deer, particularly in those areas on the mainland where densities of terrestrial ungulates are relatively low. Received: 1 July 1998 / Accepted: 23 February 1999     

Macroscopic prey remains in the winter diet of foxes Vulpes vulpes in rural Britain

• 1 The winter diet of foxes Vulpes vulpes was quantified in seven landscape types in Britain, using faecal samples from 87 sites.
• 2 Medium‐sized mammals (0.1–5.0 kg) were consistently the most important prey group in arable and pastural landscapes, occurring in 44–72% of scats and comprising 50–75% of the mass of prey ingested. Birds and small mammals (< 0.1 kg) were important secondary prey groups.
• 3 Small mammals were the most frequently recorded prey group in marginal upland (42% of scats) and upland landscapes (75%), followed by large mammals (33% and 23%, respectively). In terms of mass ingested, small mammals (38%) and large mammals (52%) were the most important prey groups in these landscapes.
• 4 In all landscapes, field voles Microtus agrestis, lagomorphs, sheep/deer and passerines/galliforms dominated their respective prey groupings.

     

Diets of crustacean zooplankton,inferred from stable carbon and nitrogen isotope analyses,in lakes with varying allochthonous dissolved organic carbon content

We used stable carbon and nitrogen isotope analyses to estimate the relative proportions of three putative food sources (1) algae, (2) allochthonous organic matter (but including also heterotrophic bacteria and green-sulphur bacteria having similar isotopic values) and (3) methane-oxidizing bacteria (MOB) in the diets of crustacean zooplankton in five small boreal lakes representing a gradient of dissolved organic carbon (DOC) concentration from ca. 5 to 40 mg C l⁻¹. The lakes were sampled in May, after establishment of stratification, and again in October during autumnal mixing of the water column. IsoSource mixing model outputs indicated that the proportion of algae in the diets of zooplankton was generally higher in May than in October, and that bacteria contributed to the diets of both cladocerans and copepods. Our results indicate that bacteria, especially MOB, can make an appreciable contribution to zooplankton diets in these small lakes, even in those with relatively low DOC concentrations.     

Carbon pathways to zooplankton: insights from the combined use of stable isotope and fatty acid biomarkers

1. Numerous studies have quantified the relative contribution of terrestrial‐ and phytoplankton‐derived carbon sources to zooplankton secondary production in lakes. However, few investigated the pathways along which allochthonous and autochthonous carbon (C) was actually conveyed to consumers. 2. We suggest that the combined use of fatty acid and stable isotope biomarkers could solve this issue. We conducted a field study on two oligotrophic lakes, in which primary production increased significantly between 2002 and 2004. We used modelling to estimate the contribution of terrestrial‐ and phytoplankton‐derived C to particulate organic C (POC) and zooplankton production from their δ¹³C values in 2002 and 2004. 3. According to the isotope model, phytoplankton‐derived C accounted for a major part of the POC pool in both lakes and supported more Daphnia sp. production in 2004 than in 2002. Fatty acid data revealed that increased contribution of algal‐C to Daphnia production, although common between both lakes, was achieved through C pathways that were different. In one lake, Daphnia grazed more intensively on phytoplankton, whereas in the other there was greater grazing on bacteria. In the latter case, the increased primary production resulted in greater release of algal‐derived dissolved organic C (DOC), which may have supported extra bacterial and eventually Daphnia, production. 4. This is the first study illustrating that the combination of fatty acid and stable isotope biomarkers could further our understanding of the factors controlling the relative magnitude of food webs pathways conveying organic matter to zooplankton.     

Untangling the complex issue of dissolved organic carbon uptake: a stable isotope approach

1. We estimated uptake of stream water dissolved organic carbon (DOC) through a whole-stream addition of a ¹³C-DOC tracer coupled with laboratory measurements of bioavailability of the tracer and stream water DOC.
2. The tracer, a leachate of ¹³C-labelled tree tissues, was added to the head waters of White Clay Creek, Pennsylvania, U.S.A., over a 2-h period and followed 1.27 km downstream to generate mass transfer coefficients for DOC lability classes within the tracer.
3. From the longitudinal ¹³C uptake curve, we resolved labile and semi-labile DOC classes within the ¹³C-DOC tracer comprising 82% and 18% of the tracer respectively.
4. Plug-flow laboratory bioreactors colonized and maintained with stream water were used to determine the concentration of stream water DOC fractions that had a similar lability to the labile and semi-labile classes within the tracer and we assumed that stream water DOC and tracer DOC with comparable lability fractions in the bioreactors behaved similarly in the stream, i.e. they had the same mass transfer coefficients.
5. A small fraction (8.6%) of the stream water DOC was labile, travelling 238 m downstream before being taken up. The remaining bioavailable stream water DOC was semi-labile and transported 4.5 km downstream before being taken up. These uptake lengths suggest that the labile DOC is an energy source within a stream reach, while the semi-labile DOC is exported out of the reach to larger rivers and the downstream estuary, where it may provide energy for marine microbial communities or simply be exported to the oceans.     

Host length,age, diet,parasites and stable isotopes as predictors of yellow perch (<Emphasis Type="Italic">Perca flavescens</Emphasis>Mitchill), trophic status in nutrient poor Canadian Shield lakes

We used data from parasites and stable isotopes of yellow perch, Perca flavescens, to determine trophic status in four small Canadian Shield lakes as parasites allow the identification of both prey and non-prey dietary components in the host's community. Stable C isotope ratios for all perch ranged from –34 to –19 while stable N isotope ratios ranged from 4.5 to 12.5. These ranges are larger than those observed in many other fish species. Perch age was the most significant predictor of stable C isotope ratio and perch parasite fauna was the most significant predictor of stable N ratios. In particular, parasite fauna indicative of zooplanktivorous or piscivorous perch were most accurate for predicting fish trophic position and thus stable isotope ratio. Fish length and age showed fewer significant relationships with isotope ratios than parasite infracommunity or diet and suggests that trophic category for perch cannot always be predicted based on size.