Quantitative approaches to the analysis of stable isotope food web data

Ecologists use stable isotopes (delta13C, delta15N) to better understand food webs and explore trophic interactions in ecosystems. Traditionally, delta13C vs. delta15N bi-plots have been used to describe food web structure for a single time period or ecosystem. Comparisons of food webs across time and space are increasing, but development of statistical approaches for testing hypotheses regarding food web change has lagged behind. Here we present statistical methodologies for quantitatively comparing stable isotope food web data. We demonstrate the utility of circular statistics and hypothesis tests for quantifying directional food web differences using two case studies: an arthropod salt marsh community across a habitat gradient and a freshwater fish community from Lake Tahoe, USA, over a 120-year time period. We calculated magnitude and mean angle of change (theta) for each species in food web space using mean delta13C and delta15N of each species as the x, y coordinates. In the coastal salt marsh, arthropod consumers exhibited a significant shift toward dependence on Spartina, progressing from a habitat invaded by Phragmites to a restored Spartina habitat. In Lake Tahoe, we found that all species from the freshwater fish community shifted in the same direction in food web space toward more pelagic-based production with the introduction of nonnative Mysis relicta and onset of cultural eutrophication. Using circular statistics to quantitatively analyze stable isotope food web data, we were able to gain significant insight into patterns and changes in food web structure that were not evident from qualitative comparisons. As more ecologists incorporate a food web perspective into ecosystem analysis, these statistical tools can provide a basis for quantifying directional food web differences from standard isotope data.     

Invasive cordgrass modifies wetland trophic function

Vascular plants strongly control belowground environments in most ecosystems. Invasion by vascular plants in coastal wetlands, and by cordgrasses (Spartina spp.) in particular, are increasing in incidence globally, with dramatic ecosystem-level consequences. We examined the trophic consequences of invasion by a Spartina hybrid (S. alterniflora x S. foliosa) in San Francisco Bay (USA) by documenting differences in biomass and trophic structure of benthic communities between sediments invaded by Spartina and uninvaded sediments. We found the invaded system shifted from an algae-based to a detritus-based food web. We then tested for a relationship between diet and tolerance to invasion, hypothesizing that species that consume Spartina detritus are more likely to inhabit invaded sediments than those that consume surface algae. Infaunal diets were initially examined with natural abundance stable isotope analyses and application of mixing models, but these yielded an ambiguous picture of food sources. Therefore, we conducted isotopic enrichment experiments by providing 15N-labeled Spartina detritus both on and below the sediment surface in areas that either contained Spartina or were unvegetated. Capitellid and nereid polychaetes, and oligochaetes, groups shown to persist following Spartina invasion of San Francisco Bay tidal flats, took up 15N from labeled native and invasive Spartina detritus. In contrast, we found that amphipods, bivalves, and other taxa less tolerant to invasion consumed primarily surficial algae, based on 13C enrichment experiments. Habitat (Spartina vs. unvegetated patches) and location of detritus (on or within sediments) did not affect 15N uptake from detritus. Our investigations support a "trophic shift" model for ecosystem response to wetland plant invasion and preview loss of key trophic support for fishes and migratory birds by shifting dominance to species not widely consumed by species at higher trophic levels.     

Hagfish in the New Zealand fjords are supported by chemoautotrophy of forest carbon

Forest litter is often considered to be a minor energy source to marine communities due to its refractory nature. Large volumes of forest litter are deposited in the New Zealand fjords, and likely recycled into available energy by microbial activity. In this study we used evidence from stable isotope analyses to test whether recycled carbon from chemoautotrophs was an important contributor to the diet of hagfish (Eptatretus cirrhatus). We then analyzed fatty acid biomarkers from the chemoautotrophic clam Solemya parkinsoni and E. cirrhatus to further discriminate the contribution of marine, terrestrial, and chemoautotrophic sources. Bulk isotopic signatures of E. cirrhatus varied considerably (delta13C, from -29.2 per thousand to -16.7 per thousand; delta15N, from -2.8 per thousand to +15.5 per thousand; delta34S, from -21.7 per thousand to +16.7 per thousand) and indicated that a significant percentage of organic matter (38-51%) originated from chemoautotrophs (delta13C, -31.3 per thousand +/- 0.1 per thousand [mean +/- SE]; delta15N, -5.7 per thousand +/- 0.2 per thousand; delta34S, -32.per thousand +/- 3.8 per thousand). Fatty acid biomarkers were depleted in 13C, particularly cis-vaccenic acid (18:1omega7: delta13C, -39.0 per thousand) indicating specific microbial origins of carbon. A high proportion of forest litter in sediments, coupled with isotopic and fatty acid biomarker results, indicates that terrestrial organic matter is a dominant contributor to this marine benthic system. This study demonstrates a clear linkage between terrestrial and marine ecological processes.     

Mercury bioaccumulation in arthropods from typical community habitats in a zinc-smelting area

This study assessed the enrichment of mercury in the food web from the different community habitats in a zinc-smelting area of China. We used a nitrogen stable isotope technique to analyze trophic level relationships among arthropods and found that the first trophic level consisted of plants in the different community habitats, the second trophic level consisted of herbivores such as locusts and grasshoppers (primary consumers), and the third trophic level included spiders and mantes (secondary consumers). Mercury enrichment in the primary consumers was not evident, but enrichment in arthropods of the third trophic level was significant. The average of enrichment coefficients in spiders and mantes was greater than 1. The δ¹⁵N values indicated that mercury concentrations accumulated from primary producers to top carnivorous arthropods increased. In this zinc-smelting area, the biological amplification of mercury in the food web is significant. It is reasonable to assume that humans, located at the top of the food chain, are exposed to biomagnified levels of mercury.     

Effects of Invasive Cordgrass on Presence of Marsh Grassbird in an Area where It Is Not Native

The threatened Marsh Grassbird (Locustella pryeri) first appeared in the salt marsh in east China after the salt marsh was invaded by cordgrass (Spartina alterniflora), a non‐native invasive species. To understand the dependence of non‐native Marsh Grassbird on the non‐native cordgrass, we quantified habitat use, food source, and reproductive success of the Marsh Grassbird at the Chongming Dongtan (CMDT) salt marsh. In the breeding season, we used point counts and radio‐tracking to determine habitat use by Marsh Grassbirds. We analyzed basal food sources of the Marsh Grassbirds by comparing the δ¹³C isotope signatures of feather and fecal samples of birds with those of local plants. We monitored the nests through the breeding season and determined the breeding success of the Marsh Grassbirds at CMDT. Density of Marsh Grassbirds was higher where cordgrass occurred than in areas of native reed (Phragmites australis) monoculture. The breeding territory of the Marsh Grassbird was composed mainly of cordgrass stands, and nests were built exclusively against cordgrass stems. Cordgrass was the major primary producer at the base of the Marsh Grassbird food chain. Breeding success of the Marsh Grassbird at CMDT was similar to breeding success within its native range. Our results suggest non‐native cordgrass provides essential habitat and food for breeding Marsh Grassbirds at CMDT and that the increase in Marsh Grassbird abundance may reflect the rapid spread of cordgrass in the coastal regions of east China. Our study provides an example of how a primary invader (i.e., cordgrass) can alter an ecosystem and thus facilitate colonization by a second non‐native species. Efectos de Spartina alterniflora Invasora Sobre Locustella pryeri en un Área Donde No Es Nativa     

Mercury bioaccumulation and trophic transfer in sympatric snapper species from the Gulf of Mexico.

Consumption of marine fish is a major route of toxic methyl mercury (MeHg) exposure to ocean apex predators and human populations. Here we explore the influence of trophic structure on total mercury (Hg) accumulation in red snapper (RS, Lutjanus campechanus) and gray snapper (GS, Lutjanus griseus) from the coastal Louisiana region of the Gulf of Mexico, west of the Mississippi River. The objectives of this investigation were to: (1) determine the effectiveness of the use of offshore recreational fishing charter boats and marinas as sources of fish samples and (2) compare species differences in Hg bioaccumulation, trophic position, and carbon sources. Our data show that length-normalized Hg concentrations (> or = 97% as MeHg in tissue of both species) were 230% greater in GS in comparison to RS collected from the same general area. Stable C and N isotope signatures (delta15N and delta13C) indicate that GS occupy a slightly higher trophic position (approximately 30% of one trophic position higher) on the Gulf food web in comparison to RS and that GS appear to incorporate higher trophic positioned prey, continually and at smaller sizes. Mercury was strongly correlated with combined delta15N and delta13C in pooled species data, arguing that most of the substantial difference in Hg bioaccumulation between RS and GS can be explained by modest differences in their trophic position and, to a lesser degree, carbon sources, which had low variation and high overlap among species. These observations demonstrate that even minor to moderate differences in trophic position and food habits in sympatric species can create relatively large differences in bioaccumulation regimes and underscores the importance of quantitative characterization of trophic structure in marine MeHg bioaccumulation studies.     

Distribution of foraging habitats of male loggerhead turtles (Caretta caretta) as revealed by stable isotopes and satellite telemetry

Most studies on the foraging ecology of loggerhead turtles (Caretta caretta) have focused on adult females and juveniles. Little is known about the foraging patterns of adult male loggerheads. We analyzed tissues for carbon and nitrogen stable isotopes (δ¹³C and δ¹⁵N) from 29 adult male loggerheads tracked with satellite transmitters from one breeding area in Florida, USA, to evaluate their foraging habitats in the Northwest Atlantic (NWA). Our study revealed large variations in δ¹³C and δ¹⁵N and a correlation between both δ¹³C and δ¹⁵N and the latitude to which the loggerheads traveled after the mating season, thus reflecting a geographic pattern in the isotopic signatures. Variation in δ¹³C and δ¹⁵N can be explained by differences in food web baseline isotopic signatures rather than differences in loggerhead trophic levels. Stable isotope analysis may help elucidate residency and migration patterns and identify foraging sea turtle subpopulations in the NWA due to the isotopically distinct habitats used by these highly migratory organisms.     

Nitrous oxide nitrification and denitrification 15N enrichment factors from Amazon forest soils.

The isotopic signatures of 15N and 18O in N2O emitted from tropical soils vary both spatially and temporally, leading to large uncertainty in the overall tropical source signature and thereby limiting the utility of isotopes in constraining the global N2O budget. Determining the reasons for spatial and temporal variations in isotope signatures requires that we know the isotope enrichment factors for nitrification and denitrification, the two processes that produce N2O in soils. We have devised a method for measuring these enrichment factors using soil incubation experiments and report results from this method for three rain forest soils collected in the Brazilian Amazon: soil with differing sand and clay content from the Tapajos National Forest (TNF) near Santarém, Pará, and Nova Vida Farm, Rond?nia. The 15N enrichment factors for nitrification and denitrification differ with soil texture and site: -111 per thousand +/- 12 per thousand and -31 per thousand +/- 11 per thousand for a clay-rich Oxisol (TNF), -102 per thousand +/- 5 per thousand and -45 per thousand +/- 5 per thousand for a sandier Ultisol (TNF), and -10.4 per thousand +/- 3.5 per thousand (enrichment factor for denitrification) for another Ultisol (Nova Vida) soil, respectively. We also show that the isotopomer site preference (delta15Nalpha - delta15Nbeta, where alpha indicates the central nitrogen atom and beta the terminal nitrogen atom in N2O) may allow differentiation between processes of production and consumption of N2O and can potentially be used to determine the contributions of nitrification and denitrification. The site preferences for nitrification and denitrification from the TNF-Ultisol incubated soils are: 4.2 per thousand +/- 8.4 per thousand and 31.6 per thousand +/- 8.1 per thousand, respectively. Thus, nitrifying and denitrifying bacteria populations under the conditions of our study exhibit significantly different 15N site preference fingerprints. Our data set strongly suggests that N2O isotopomers can be used in concert with traditional N2O stable isotope measurements as constraints to differentiate microbial N2O processes in soil and will contribute to interpretations of the isotopic site preference N2O values found in the free troposphere.     

Microscale variations of food web functioning within a rocky shore invertebrate community

The assessment of relevant spatial scales at which ecological processes occur is of special importance for a thorough understanding of ecosystem functioning. In coastal ecosystems, the variability of trophic interactions has been studied at different spatial scales, but never at scales from centimetres to metres. In the present study, we investigated the link between habitat structure and small-scale variability of food web functioning on intertidal boulder field ecosystems. Two microhabitats, boulder-top and boulder-bottom, were considered, and the trophic ecology of invertebrate consumers was studied using stable isotope tracers. We found for two of the main suspension feeders of northern Atlantic rocky shores (the sponges Halichondria panicea and Hymeniacidon sanguinea) consistent ¹⁵N enrichment for individuals sampled under boulders, suggesting that these consumers relied on different trophic resource according to the microhabitat inhabited, at a centimetre scale. The high δ¹⁵N signatures found underneath boulders suggested higher use of highly decomposed organic matter in this microhabitat. The isotopic difference between the two microhabitats decreased in higher trophic level consumers, which likely foraged at a spatial scale including both microhabitats. Finally, our results reveal that in highly heterogeneous habitats such as boulder fields, trophic interactions are likely to vary strongly in space, which should be considered in future researches. The link between habitat physical structure and food web variability might also contribute to the high biological diversity characterizing heterogeneous ecosystems.     

Spatial and temporal shifts in stable isotope values of the bottom-dwelling shrimp<Emphasis Type="Italic"> Nauticaris marionis</Emphasis> at the sub-Antarctic archipelago

Spatial and temporal dynamics of carbon and nitrogen stable isotope signatures of the bottom-dwelling caridean shrimp Nauticaris marionis were measured during April and May between 1984 and 2000 in the vicinity of Marion Island (the Prince Edward Islands, Southern Ocean). There was one trophic-level enrichment in bulk ¹⁵N and ¹³C signatures between small (<20 mm long) and large (>20 mm) specimens of N. marionis, suggesting distinct trophic differentiation among major shrimp size groups. Both ¹⁵N and ¹³C values of N. marionis increased with the depth, reflecting changes in their diet. There were no clear temporal trends in bulk ¹⁵N signatures of N. marionis. However, compound-specific ¹⁵N measurements of amino acids indicated that N. marionis from the inter-island realm occupied the trophic level of second order carnivores, while similarly sized shrimps in the near-shore realm were at the trophic level of first order carnivores. Compound-specific measurements also identified a change in the source of inorganic nitrogen at the base of the food web between the inter-island and near-shore realms. In contrast to the bulk ¹⁵N values, a significant shift in bulk ¹³C values of N. marionis was observed between 1984 and more recent years. This temporal change appears to be linked to changes in the overall productivity of the Prince Edward Island inter-island system, which could be linked to global climate change.Communicated by J.P. Thorpe, Port Erin     

Can stable isotope ratios provide for community-wide measures of trophic structure?

Stable isotope ratios (typically of carbon and nitrogen) provide one representation of an organism's trophic niche and are widely used to examine aspects of food web structure. Yet stable isotopes have not been applied to quantitatively characterize community-wide aspects of trophic structure (i.e., at the level of an entire food web). We propose quantitative metrics that can be used to this end, drawing on similar approaches from ecomorphology research. For example, the convex hull area occupied by species in delta13C-delta15N niche space is a representation of the total extent of trophic diversity within a food web, whereas mean nearest neighbor distance among all species pairs is a measure of species packing within trophic niche space. To facilitate discussion of opportunities and limitations of the metrics, we provide empirical and conceptual examples drawn from Bahamian tidal creek food webs. These examples illustrate how this methodology can be used to quantify trophic diversity and trophic redundancy in food webs, as well as to link individual species to characteristics of the food web in which they are embedded. Building from extensive applications of stable isotope ratios by ecologists, the community-wide metrics may provide a new perspective on food web structure, function, and dynamics.     

Using stable isotopes to reveal shifts in prey consumption by generalist predators.

The effectiveness of generalist predators in biological control may be diminished if increased availability of alternative prey causes individual predators to decrease their consumption of crop pests. Farming practices that enhance densities of microbidetritivores in the detrital food web can lead to increased densities of generalist predators that feed on pest species. The ability to predict the net biocontrol impact of increased predator densities depends upon knowing the extent to which individual predators may shift to detrital prey and feed less on crop pests when prey of the detritus-based food web are more abundant. We addressed this question by comparing ratios of stable isotopes of carbon (delta13C) and nitrogen (delta15N) in generalist ground predators and two types of prey (crop pests and microbidetritivores) in replicated 8 x 8 m cucurbit gardens subjected to one of two treatments: a detrital subsidy or no addition of detritus (control). Small sheet-web spiders (Linyphiidae) and small wolf spiders (Lycosidae) had delta13C values similar to those of Collembola in both the detrital and control treatments, indicating that small spiders belong primarily to the detrital food web. In control plots the larger generalist predators had delta13C values similar to those of the major insect pests, consistent with their known effectiveness as biocontrol agents. Adding detritus may have caused delta13C of one species of large wolf spider to shift toward that of the microbi-detritivores, although evidence is equivocal. In contrast, another large wolf spider displayed no shift in delta13C in the detrital treatment. Thus, stable isotopes revealed which generalist predators will likely continue to feed on pest species in the presence of greater densities of alternative prey.     

Nutrient subsidies to belowground microbes impact aboveground food web interactions

Historically, terrestrial food web theory has been compartmentalized into interactions among aboveground or belowground communities. In this study we took a more synthetic approach to understanding food web interactions by simultaneously examining four trophic levels and investigating how nutrient (nitrogen and carbon) and detrital subsidies impact the ability of the belowground microbial community to alter the abundance of aboveground arthropods (herbivores and predators) associated with the intertidal cord grass Spartina alterniflora. We manipulated carbon, nitrogen, and detrital resources in a field experiment and measured decomposition rate, soil nitrogen pools, plant biomass and quality, herbivore density, and arthropod predator abundance. Because carbon subsidies impact plant growth only indirectly (microbial pathways), whereas nitrogen additions both directly (plant uptake) and indirectly (microbial pathways) impact plant primary productivity, we were able to assess the effect of both belowground soil microbes and nutrient availability on aboveground herbivores and their predators. Herbivore density in the field was suppressed by carbon supplements. Carbon addition altered soil microbial dynamics (net potential ammonification, litter decomposition rate, DON [dissolved organic N] concentration), which limited inorganic soil nitrogen availability and reduced plant size as well as predator abundance. Nitrogen addition enhanced herbivore density by increasing plant size and quality directly by increasing inorganic soil nitrogen pools, and indirectly by enhancing microbial nitrification. Detritus adversely affected aboveground herbivores mainly by promoting predator aggregation. To date, the effects of carbon and nitrogen subsidies on salt marshes have been examined as isolated effects on either the aboveground or the belowground community. Our results emphasize the importance of directly addressing the soil microbial community as a factor that influences aboveground food web structure by affecting plant size and aboveground plant nitrogen.     

Food web structure of a restored macroalgal bed in the eastern Korean peninsula determined by C and N stable isotope analyses

Loss of macroalgae habitats has been widespread on rocky marine coastlines of the eastern Korean peninsula, and efforts for restoration and creation of macroalgal beds have increasingly been made to mitigate these habitat losses. Deploying artificial reefs of concrete pyramids with kelps attached has been commonly used and applied in this study. As a part of an effort to evaluate structural and functional recovery of created and restored habitat, the macroalgal community and food web structure were studied about a year after the establishment of the artificial macroalgal bed, making comparisons with nearby natural counterparts and barren ground communities. Dominant species, total abundance, and community structure of macroalgal assemblage at the restored macroalgal bed recovered to the neighboring natural bed levels during the study period. The main primary producers (phytoplankton and macroalgae) were isotopically well separated. δ¹³C and δ¹⁵N values of consumers were very similar between restored and natural beds but varied greatly among functional feeding groups. The range of consumer δ¹³C was as wide as that of primary producers, indicating the trophic importance of both producers. There was a stepwise trophic enrichment in δ¹⁵N with increasing trophic level. A comparison of isotope signatures between primary producers and consumers showed that, while suspension feeders are highly dependent on pelagic sources, invertebrates of other feeding guilds and fishes mainly use macroalgae-derived organic matter as their ultimate nutritional sources in both macroalgal beds, emphasizing the high equivalency of trophic structure between both beds. Isotopic signatures of a few mollusks and sea urchins showed that they use different dietary items in macroalgal-barren grounds compared with macroalgal beds, probably reflecting their feeding plasticity according to the low macroalgal biomass. However, isotopic signatures of most of the consumers at the barren ground were consistent with those at the macroalgal beds, supporting the important trophic role of drifting algae. Our results revealed the recoveries of the macroalgal community and trophic structure at the restored habitat. Further studies on colonization of early settlers and the following succession progress are needed to better understand the process and recovery rate in the developing benthic community.     

Kelp as a trophic resource for marine suspension feeders: a review of isotope-based evidence

Kelp forests are enormously productive, and they and adjacent habitats support large populations of suspension feeders. What do these suspension feeders eat? Intuitively, we might expect that kelp primary production is a key form of trophic support for these animals. Indeed, a large and growing number of studies using carbon stable isotope data, typically collected over short time periods, have asserted that detritus from kelps is an important, and in some cases the main, food source for coastal benthic suspension feeders. This view has been incorporated into several textbooks and review papers covering kelp forest ecosystems, and loss of trophic support for benthic suspension feeders is now often invoked as an ecosystem consequence of top-down or other impacts on kelp forests. More direct evidence, however, suggests that these animals mainly eat phytoplankton and, in some cases, bacteria or zooplankton. Because isotope values of pure coastal phytoplankton, uncontaminated with detritus, are difficult to obtain, present studies have largely relied on single measurements from offshore environments or from the literature, which typically reflects offshore values. We review the evidence showing that phytoplankton isotope values can, and are expected to, vary widely in coastal waters and that inshore phytoplankton may often be enriched in ¹³C compared to offshore phytoplankton. This unaccounted-for variation may have systematically biased the results of such trophic studies toward finding large contributions of kelp detritus to suspension-feeder diets. We review some key stable isotope studies and put forth evidence for alternative explanations of the isotope patterns presented. Finally, we make recommendations for future isotope studies and describe several approaches for progress in this area. New techniques, particularly flow cytometry and compound-specific stable isotope analysis, provide ways to shed light on this interesting and important ecological issue.     

Incorporating temporally dynamic baselines in isotopic mixing models

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

Nitrogen inputs promote the spread of an invasive marsh grass.

Excess nutrient loading and large-scale invasion by nonnatives are two of the most pervasive and damaging threats to the biotic and economic integrity of our estuaries. Individually, these are potent forces, but it is important to consider their interactive impacts as well. In this study we investigated the potential limitation of a nonnative intertidal grass, Spartina alterniflora, by nitrogen (N) in estuaries of the western United States. Nitrogen fertilization experiments were conducted in three mud-flat habitats invaded by S. alterniflora in Willapa Bay, Washington, USA, that differed in sediment N. We carried out parallel experiments in San Francisco Bay, California, USA, in three habitats invaded by hybrid Spartina (S. alterniflora x S. foliosa), in previously unvegetated mud flat, and in native S. foliosa or Salicornia virginica marshes. We found similar aboveground biomass and growth rates between habitats and estuaries, but end-of-season belowground biomass was nearly five times greater in San Francisco Bay than in Willapa Bay. In Willapa Bay, aboveground biomass was significantly correlated with sediment N content. Addition of N significantly increased aboveground biomass, stem density, and the rate of spread into uninvaded habitat (as new stems per day) in virtually all habitats in both estuaries. Belowground biomass increased in Willapa Bay only, suggesting that belowground biomass is not N limited in San Francisco Bay due to species differences, N availability, or a latitudinal difference in the response of Spartina to N additions. The relative impact of added N was greater in Willapa Bay, the estuary with lower N inputs from the watershed, than in San Francisco Bay, a highly eutrophic estuary. Nitrogen fertilization also altered the competitive interaction between hybrid Spartina and Salicornia virginica in San Francisco Bay by increasing the density and biomass of the invader and decreasing the density of the native. There was no significant effect of N on the native, Spartina foliosa. Our results indicate that excess N loading to these ecosystems enhances the vulnerability of intertidal habitats to rapid invasion by nonnative Spartina sp.     

Whole-lake dissolved inorganic 13C additions reveal seasonal shifts in zooplankton diet

Sustained whole-lake additions of 13C-enriched dissolved inorganic carbon (DIC), intended to increase experimentally the delta13C of DIC in the epilimnion of a small lake with high dissolved organic carbon (DOC), were made during three seasonal periods (spring, summer, and autumn). Coupled with carbon and nitrogen stable isotope analysis of zooplankton and several of their putative food sources, these additions were used to investigate seasonal changes in the relative contributions of different food sources to zooplankton diet in the lake. Four main potential food sources were considered: phytoplankton, heterotrophic bacteria (HB), methanotrophic bacteria (MOB), and green sulfur bacteria (GSB). Because the number of potential food sources exceeded the number of isotopes analyzed, a computer program (IsoSource) was used to estimate the range of possible contributions of the various food sources. During all three periods the added inorganic 13C quickly increased the epilimnetic DIC delta13C by between 18 per thousand and 21 per thousand above the initial value of approximately -21 per thousand. This 13C enrichment of DIC was rapidly transmitted to the particulate organic matter (POM), which included photosynthetic phytoplankton. In spring and summer, delta13C of both adult and juvenile Daphnia increased by approximately 10 per thousand, indicating that Daphnia utilized autochthonous carbon. However, this 13C labeling of Daphnia was not so obvious during the autumn period, when their delta13C generally decreased. According to the IsoSource model outputs based on both delta13C and delta15N values, Daphnia utilized all four potential food source types during spring, summer, and autumn, but in different proportions. The possible contribution of phytoplankton to Daphnia diet was substantial (25-71%) in all seasons. The possible contributions of the bacterial food sources were more variable. The possible contribution of GSB was minor (0-20%) at all times and negligible in autumn. The possible contribution of HB was higher but very variable. Methanotrophic bacteria always made a significant contribution to Daphnia diet and were likely the single most important food source in autumn. Since both HB and MOB in this high-DOC lake probably depend largely on allochthonous organic carbon, our results highlight the seasonal variability in the potential importance of ecosystem subsidies in lake food webs.     

Sexual segregation in distribution,diet and trophic level of seabirds: insights from stable isotope analysis

Considerable attention has focused on inter- and intraspecific variation in trophic niches of marine predators. Although this has revealed evidence for sexual segregation in distribution in some species, few studies have been able to address sex-related dietary specialisation. Stable isotope analysis of blood cells collected from albatrosses and petrels at South Georgia during chick-rearing indicated a difference in δ¹³C, suggesting that females fed to the north of males, only in two species with male-biased sexual size dimorphism; in no species did sexes differ in trophic level (δ¹⁵N). Based on a wider review, significant differences between sexes in isotope signatures were much more common in seabirds during the pre-laying or breeding than the nonbreeding period, presumably reflecting greater between-sex partitioning of resources when foraging ranges are more constrained and competition is greater. Sex differences, or their absence, were usually consistent across successive stages during the pre-laying and breeding periods, but not necessarily year-round nor between populations. Significant differences in isotope signatures between males and females were extremely rare in monomorphic species, suggesting a link between sexual size dimorphism and segregation in diet or distribution. Among the Southern Ocean albatrosses, sex differences in δ¹³C suggested the underlying mechanism was related to habitat specialisation, whereas in other size-dimorphic taxa (both male- and female-biased), sex differences were more common in δ¹⁵N than δ¹³C and therefore more consistent with size-mediated competitive exclusion or dietary specialisation.     

Incorporation of plant carbon into the soil animal food web of an arable system

We used stable isotopes to examine the incorporation of plant carbon into the belowground food web of an agricultural system. Plots were established and planted with maize (Zea mays) in a rye field (Secale cereale) near G?ttingen (northern Germany) in May 1999. In October 1999, April 2000, and October 2000, meso- and macrofauna and maize and rye litter were collected in each plot and analyzed for 13C and 15N content. 15N signatures suggested that the soil animal species analyzed span three trophic levels with the trophic position of species varying little in time. The species investigated formed a continuum from primary to secondary decomposers to predators. On average, predator species differed from primary and secondary decomposers by 3.9 sigma15N suggesting that they fed on a mixed diet of both decomposer groups. The combined analysis of 13C and 15N signatures allowed us to identify links between prey and consumer species. In October 1999, shortly after maize residues had been incorporated into the plots, maize-born carbon was present in each of the animal species investigated, including top predators. The incorporation of maize carbon into the belowground food web increased during the following 12 months but the concentration of maize-born carbon never exceeded 50% in any of the species. Furthermore, the ranks of the incorporation of maize-born carbon of the species changed little. The results suggest that the belowground food web relies heavily on carbon originating from plant residues from before the recent two growing seasons. In most species the amount of maize-born carbon increased continuously; however, in some species it decreased during winter, suggesting that these species switched to a diet based more on C3 plants during winter, or predominantly metabolized carbon incorporated during the last growing season. The study documents that the combined analysis of 13C and 15N signatures in soil invertebrate species, after replacement of C3 by C4 plants, is a powerful tool to better understand the structure of the belowground food web and the flux of carbon through it.