Dietary Routing Research Articles

Trophic positions of soil microarthropods in forests increase with elevation, but energy channels remain unchanged

AbstractMountain forests are at risk as the consequences of climate change will likely lead to altered tree species boundaries. Characterizing food webs along elevation gradients in primary forests may help to predict the potential consequences of such changes, for example with regard to the decomposition of dead organic matter. Here, for the first time, we studied trophic variations in two species‐rich microarthropod taxa, Collembola and Oribatida, along an elevation gradient of primary forest at Changbai Mountain, China. Samples were taken at seven elevations of 150‐m elevational difference between 800 and 1700 m. At each elevation, Collembola and Oribatida were extracted from litter samples of eight subplots. We applied three state‐of‐the‐art methods to elucidate trophic positions and basal resource use at community level: Bulk stable isotope analysis of nitrogen (Δ15Nbulk) and carbon (Δ13Cbulk), compound‐specific stable isotope analysis of amino acids (CSIA‐AA), and dietary routing of neutral lipid fatty acids (NLFAs). Trophic positions calculated using Δ15Nbulk and CSIA‐AA (TPCSIA) in both taxa increased similarly with elevation by about half and one third of a trophic position, respectively. Stable isotope mixing models and linear discriminant analysis bootstrapping using δ13C of essential amino acids indicated fungi as the most important resource at all elevations for both taxa. Also, proportions of marker NLFAs changed little across elevations in both taxa; overall high proportions of linoleic acid indicated high fungal contributions, but in Collembola the contribution of bacterial markers was generally higher than in Oribatida. Δ13Cbulk did not respond linearly to the elevation gradient; however, changes in elevation differed between Collembola and Oribatida. A strong linear relationship between δ15N of phenylalanine and δ15N of litter indicated litter as the basis of energy channels in both taxa. Overall, food web functioning likely changes with changing forest types along elevation gradients, with microarthropods switching from feeding closer to the base of the food web at lower elevations to feeding at higher trophic levels at higher elevations, potentially compromising their role in litter decomposition and nutrient cycling.

Effects of resource quality on the fitness of collembola fed single and mixed diets from the green and brown food chain

The effect of food resources, varying in C/N ratio, lipid and ω3 polyunsaturated fatty acid (ω3-PUFA) content, on the fitness of three Collembola species (the euedaphic Protaphorura fimata, hemiedpahic Folsomia candida and epedaphic Lepidocyrtus violaceus) was investigated. Dietary routing of fatty acids served as an indicator for resource consumption and assimilation. Laboratory feeding experiments were performed with single and mixed diets made from algae, cyanobacteria, fungi, soil bacteria or plant roots and leaves. Collembola exhibited species-specific responses in biomass and survival rate to the diets differing in quality, reflecting adaptations to their soil environment. Generally, wide dietary C/N ratios had a negative impact, with roots especially unfavorable, while the dependence on ω3-PUFAs varied. Mixing diets did mostly result in intermediate results where high-quality resources compensate for low-quality resources, with only mixes of resources favoured by Collembola in the wild showing synergistic positive effects. The results suggest that most resources of soil detritivores, irrespective of green or brown food chain, represent a non-optimal diet, with clear adaptations depending on habitat and the potential to truly exploit any resource mostly exhibited in euedaphic species.

Proteomic profile of bone "collagen" extracted for stable isotopes: Implications for bulk and single amino acid analyses.

Protein studies in archaeology and paleontology have been dominated by stable isotope studies to understand diet and trophic levels, but recent applications of proteomic techniques have resulted in a more complete understanding of protein diagenesis than stable isotopes alone. In stable isotope analyses, samples are retained or discarded based on their properties. Proteomics can directly determine what proteins are present within the sample and may be able to allow previously discarded samples to be analyzed. Protein samples that had been previously analyzed for stable isotopes, including those with marginal and poor sample quality, were characterized by liquid chromatography/mass spectrometry using an LTQ Orbitrap Velos mass spectrometer after separation on a Dionex Ultimate 3000 LC system. Data were analyzed using MetaMorpheus and custom R scripts. We found a variety of proteins in addition to collagen, although collagen I was found in the majority of the samples (most samples >80%). We also found a positive correlation between total deamidation and wt% N, suggesting that deamidation may impact the overall nitrogen signal in bulk analyses. The amino acid profiles of samples, including those of marginal or poor stable isotope quality, reflect the expected collagen I percentages, allowing their use in single amino acid stable isotope analyses. All the samples regardless of quality were found to have high concentrations of collagen I, making interpretations of dietary routing based on collagen I reasonably valid. The amino acid profiles on the marginal and poor samples reflect an expected collagen I profile and allow these samples to be recovered for single amino acid analyses.

Nematodes as Important Source for Omega-3 Long-Chain Fatty Acids in the Soil Food Web and the Impact in Nutrition for Higher Trophic Levels

PUFA (polyunsaturated fatty acids) content of food sources, in particular 3 long-chain PUFA with three or more double bonds, are considered essential for growth, reproduction, and neural development of higher animals. Surprisingly, and in contrast to aquatic ecosystems, 3 long-chain PUFA seem not widely available in terrestrial food webs. Far-reaching ideas indeed proclaim aquatic ecosystems as the principal source of these long-chain PUFA in the whole biosphere, including inhabitants of terrestrial ecosystems. Interestingly, de novo synthesis of 3 long-chain PUFA, which requires the presence of Δ12 (fat-2) and 3 (fat-1) desaturases absent in vertebrates, has been observed in nematodes, such as Caenorhabditis elegans. This raises the question if nematodes or other soil invertebrates present an important trophic link offering substantial supply in these valuable nutritional compounds in terrestrial food webs. This work followed the dietary routing of fatty acids of different C. elegans strains, including mutants defective in the PUFA biosynthesis, to two omnivorous Collembola species, Folsomia candida and Protaphorura fimata. The laboratory approach comprised microcosms offering binary links under various feeding conditions and analyzed growth, fecundity and dietary preference of consumers. Collembola did not prefer individual C. elegans strains as food source but they clearly reflected the PUFA-richness or -poorness of their nematode prey in their neutral lipid fraction. Moreover, Collembola did benefit from 3 long-chain PUFA rich diet, as shown by significant weight gain and increased number of laid eggs. Interestingly, the comparatively high PUFA-content of Collembola’s phospholipid fraction remained unchanged, even in response to almost PUFA-depleted nematode prey, suggesting that these Collembola species also possess the metabolic capability to de novo synthesize PUFA, including 3 long-chain forms. These findings broaden the basis of long-chain PUFA sources in terrestrial food webs and question the impact of aquatic ecosystems as principal source.

Fatty Acid 13C Isotopologue Profiling Provides Insight into Trophic Carbon Transfer and Lipid Metabolism of Invertebrate Consumers.

Fatty acids (FAs) are useful biomarkers in food web ecology because they are typically assimilated as a complete molecule and transferred into consumer tissue with minor or no modification, allowing the dietary routing between different trophic levels. However, the FA trophic marker approach is still hampered by the limited knowledge in lipid metabolism of the soil fauna. This study used entirely labelled palmitic acid (13C16:0, 99 atom%) as a tracer in fatty acid metabolism pathways of two widespread soil Collembola, Protaphorura fimata and Heteromurus nitidus. In order to investigate the fate and metabolic modifications of this precursor, a method of isotopologue profiling is presented, performed by mass spectrometry using single ion monitoring. Moreover, the upstream laboratory feeding experiment is described, as well as the extraction and methylation of dominant lipid fractions (neutral lipids, phospholipids) and the related formula and calculations. Isotopologue profiling does not only yield the overall 13C enrichment in fatty acids derived from the 13C labeled precursor but also produces the pattern of isotopologues exceeding the mass of the parent ion (i.e., the FA molecular ion M+) of each labeled FA by one or more mass units (M+1, M+2, M+3, etc.). This knowledge allows conclusions on the ratio of dietary routing of an entirely consumed FA in comparison to de novo biosynthesis. The isotopologue profiling is suggested as a useful tool for evaluation of fatty acid metabolism in soil animals to disentangle trophic interactions.

Isotopologue profiling enables insights into dietary routing and metabolism of trophic biomarker fatty acids

The application of fatty acids (FAs) as biomarkers in food web ecology is based on the concept of dietary routing, i.e., FA assimilation as entire molecule and transfer into consumer tissue with no or minor modification. However, the lack in knowledge in lipid metabolism of the soil fauna hampers the FA trophic marker approach. This study used labeled palmitic acid (13C16:0, 99 atom%) as tracer in lipid pathways of two common soil Collembola, Heteromurus nitidus and Protaphorura fimata. The incorporation and dietary routing of this precursor were determined by 13C:12C FA analysis (gas chromatography–combustion–isotope-ratio-monitoring), while metabolic modifications were assessed with isotopologue profiling (mass spectrometry with single ion monitoring). The 13C enrichment in FAs assigned the transfer of the labeled precursor predominantly into the neutral lipid fraction of Collembola. Thereby, isotopologue profiling demonstrated that dietary routed FAs are marginally metabolically modified. In contrast, the phospholipid fraction showed a considerable degree of FA modification, most markedly a fast degradation of palmitoleic acid. Further, de novo synthesis of linoleic acid was approved for both Collembola species. While 16:0 and 18:0, and their descendants 16:1ω7 and 18:1ω9, followed the concept of dietary routing and thus are good markers, arachidonic acid showed a different metabolism. This comprised elongation/desaturation of 13C16:0 and de novo synthesis, including 13C labeled acetyl-CoA from degradation of the precursor, indicating its utility to assign carbon flow but not as trophic marker. We propose isotopologue profiling as a valuable tool for evaluation of the lipid metabolism in small soil animals to disentangle trophic interactions.

A Simple(R) Model to Predict the Source of Dietary Carbon in Individual Consumers

Quantitative individual human diet reconstruction using isotopic data and a Bayesian approach typically requires the inclusion of several model parameters, such as individual isotopic data, isotopic and macronutrient composition of food groups, diet-to-tissue isotopic offsets and dietary routing. In an archaeological context, sparse data may hamper a widespread application of such models. However, simpler models may be proposed to address specific archaeological questions. As a consequence of the intake of marine foods, individuals from the first century ad Roman site of Herculaneum showed well-defined bone collagen radiocarbon age offsets from the expected terrestrial value. Taking as reference these radiocarbon offsets and using as model input stable isotope data (δ13C and δ15N), the performance of two Bayesian mixing model instances (routed and concentration-dependent model versus non-routed and concentration-independent) was compared to predict the carbon contribution of marine foods to bone collagen. Predictions generated by both models were in good agreement with observed values. The model with higher complexity showed only a slightly better performance in terms of accuracy and precision. This demonstrates that under similar circumstances, a simple Bayesian approach can be applied to quantify the carbon contribution of marine foods to human bone collagen.

Dietary reconstruction, mobility, and the analysis of ancient skeletal tissues: Expanding the prospects of stable isotope research in archaeology

The use of stable isotope ratio analysis in archaeology has exploded over the past few decades to the point where it is now an established tool that is routinely used to investigate questions relating to diet and mobility. Early applications focused mostly on the analysis of human skeletal tissues as a way to reconstruct major shifts in human diet, but current stable isotopic approaches have expanded to include high resolution analyses of human, animal, and plant remains, which are helping to better define the resource exploitation and management strategies that underscore changes in the human diet. In addition, stable isotopic data sets are now regularly filtered through interpretive archaeological theoretical frameworks to explore socially mediated food acquisition and consumption choices, mortuary practices, and social identity. Much work remains to be done in documenting the biological and ecological variation in the distribution of stable isotopes in ancient food webs and the mechanisms responsible for the isotopic signals observed in archaeological plant and animal tissues. Here, we identify several areas in stable isotope analysis where additional ‘first principles’ driven research would help to improve existing isotopic methods, or develop new ones, and consequently improve our ability to answer questions of archaeological significance. We consider the strengths and limitations of the application of stable isotope analysis to ancient skeletal material obtained from archaeological contexts. We also pay particular attention to nitrogen isotopic variation in ancient ecosystems, organic oxygen and hydrogen isotopes to;mixing models as a means of estimating source contributions in human diet, mobility, and isoscapes; and to how compound specific analyses may help detangle dietary routing. We conclude with a plea for greater scientific rigour and more informed use of stable isotope analyses and call for a closer integration of stable isotope analysis with other aspects of archaeological research programmes, in order to optimise the information that isotopes can provide.

Food Reconstruction Using Isotopic Transferred Signals (FRUITS): A Bayesian Model for Diet Reconstruction

Human and animal diet reconstruction studies that rely on tissue chemical signatures aim at providing estimates on the relative intake of potential food groups. However, several sources of uncertainty need to be considered when handling data. Bayesian mixing models provide a natural platform to handle diverse sources of uncertainty while allowing the user to contribute with prior expert information. The Bayesian mixing model FRUITS (Food Reconstruction Using Isotopic Transferred Signals) was developed for use in diet reconstruction studies. FRUITS incorporates the capability to account for dietary routing, that is, the contribution of different food fractions (e.g. macronutrients) towards a dietary proxy signal measured in the consumer. FRUITS also provides relatively straightforward means for the introduction of prior information on the relative dietary contributions of food groups or food fractions. This type of prior may originate, for instance, from physiological or metabolic studies. FRUITS performance was tested using simulated data and data from a published controlled animal feeding experiment. The feeding experiment data was selected to exemplify the application of the novel capabilities incorporated into FRUITS but also to illustrate some of the aspects that need to be considered when handling data within diet reconstruction studies. FRUITS accurately predicted dietary intakes, and more precise estimates were obtained for dietary scenarios in which expert prior information was included. FRUITS represents a useful tool to achieve accurate and precise food intake estimates in diet reconstruction studies within different scientific fields (e.g. ecology, forensics, archaeology, and dietary physiology).

An analysis of diet quality, how it controls fatty acid profiles, isotope signatures and stoichiometry in the malaria mosquito Anopheles arabiensis.

BackgroundKnowing the underlying mechanisms of mosquito ecology will ensure effective vector management and contribute to the overall goal of malaria control. Mosquito populations show a high degree of population plasticity in response to environmental variability. However, the principle factors controlling population size and fecundity are for the most part unknown. Larval habitat and diet play a crucial role in subsequent mosquito fitness. Developing the most competitive insects for sterile insect technique programmes requires a “production” orientated perspective, to deduce the most effective larval diet formulation; the information gained from this process offers us some insight into the mechanisms and processes taking place in natural native mosquito habitats.Methodology/Principal FindingsFatty acid profiles and de-novo or direct assimilation pathways, of whole-individual mosquitoes reared on a range of larval diets were determined using pyrolysis gas chromatograph/mass spectrometry. We used elemental analysis and isotope ratio mass spectrometry to measure individual-whole-body carbon, nitrogen and phosphorous values and to assess the impact of dietary quality on subsequent population stoichiometry, size, quality and isotopic signature. Diet had the greatest impact on fatty acid (FA) profiles of the mosquitoes, which exhibited a high degree of dietary routing, characteristic of generalist feeders. De-novo synthesis of a number of important FAs was observed. Mosquito C:N stoichiometry was fixed in the teneral stage. Dietary N content had significant influence on mosquito size, and P was shown to be a flexible pool which limited overall population size.Conclusions/SignificanceDirect routing of FAs was evident but there was ubiquitous de-novo synthesis suggesting mosquito larvae are competent generalist feeders capable of survival on diet with varying characteristics. It was concluded that nitrogen availability in the larval diet controlled teneral mosquito size and that teneral CN ratio is a sex- and species-specific fixed parameter. This finding has significant implications for overall mosquito competitiveness and environmental management.

Macronutrient-based model for dietary carbon routing in bone collagen and bioapatite

Carbon stable isotope ratios (δ 13 C), measured in human bone collagen (δ 13 Ccollagen) and bioapatite (δ 13 Cbioapatite),

Quantifying dietary macronutrient sources of carbon for bone collagen biosynthesis using natural abundance stable carbon isotope analysis

The diets of laboratory rats were isotopically and nutritionally manipulated using purified C3 and/or C4 macronutrients to investigate the routing of dietary carbon to bone collagen biosynthesis. Diets were formulated with purified proteins, carbohydrates and lipids of defined composition and natural abundance stable isotope ratios. Bulk protein and constituent amino acid delta(13)C values determined for whole diet and bone collagen provided the basis for assessing isotopic fractionation and estimating the degree of routing versus synthesis de novo of essential, non-essential and conditionally indispensable amino acids. Essential and conditionally indispensable amino acids were shown to be routed from diet to collagen with little isotopic fractionation whereas non-essential amino acids differed by up to 20 per thousand. Mathematical modelling of the relationships between macronutrient and tissue delta(13)C values provided qualitative and quantitative insights into the metabolic and energetic controls on bone collagen biosynthesis. Essential amino acids comprise 21.7 % of the carbon in collagen, defining the minimum amount of dietary carbon routing. Estimates of 42 and 28 % routing were shown for the non-essential amino acids, glycine and aspartate, respectively. In total, the routing of non-essential and conditionally indispensable amino acids was estimated to equal 29.6 % of the carbon in collagen. When the contribution of carbon from the essential amino acids is also considered, we arrive at an overall minimum estimate of 51.3 % routing of dietary amino acid carbon into bone collagen.

Dietary routing of nutrients from prey to offspring in a generalist predator: effects of prey quality

Summary Reproduction of female wolf spiders (Pardosa lugubris; Lycosidae) fed with prey of different quality was investigated. Spiders were fed either a single diet of Drosophila melanogaster (Diptera, high quality), Heteromurus nitidus (Collembola, high quality) or Folsomia candida (Collembola, toxic), or a mixed diet of D. melanogaster and H. nitidus, and of D. melanogaster and F. candida. Nutrient flow from prey into females and from females into offspring during egg production was investigated tracing carbon and nitrogen stable isotopes with prey being enriched in 13C and/or 15N. There was no benefit of mixing different high‐quality prey on female biomass and performance, contrasting with earlier results obtained with juvenile spiders. Folsomia candida was toxic for P. lugubris: females, even if fed a mixed diet also containing high‐quality prey, did not reproduce and finally died; spiders did not acquire aversion against F. candida. Tracing stable isotopes documented the incorporation of C and N from prey into females and their offspring; dietary nutrients were routed almost exclusively into egg production. Stable isotope analysis strongly supported the assumption that F. candida causes post‐ingestive physiological effects in spiders by inhibiting the incorporation of nutrients from other prey.

Carbon stable isotope fractionation and trophic transfer of fatty acids in fungal based soil food chains

Stable isotope analysis has been used as a powerful tool in food web studies in terrestrial ecosystems. In addition the occurrence and abundance of fatty acids may serve as indicator for feeding strategies of soil animals. Here we combine both approaches and investigate the fatty acid composition, δ 13C values of bulk tissues and individual fatty acids in soil organisms. The fungi Chaetomium globosum and Cladosporium cladosporioides were isotopically labelled by fructose derived from either C 3 or C 4 plants, and the fungal-feeding nematode Aphelenchoides sp. was reared on C. globosum. Fungi and nematodes were used as diet for the Collembolan Protaphorura fimata. The sugar source was fractionated differently by fungal lipid metabolism in a species-specific manner that points to a sensitivity of physiological processing to the non-random distribution of 13C/ 12C isotopes in the molecule. As a general trend stearic acid (18:0) was depleted in 13C compared to the precursor palmitic acid (16:0), whereas its desaturation to oleic acid (18:1 ω9) favoured the 13C-rich substrate. Fatty acid profiles of P. fimata varied due to food source, indicating incorporation of dietary fatty acids into Collembolan tissue. Individuals feeding on fungi had lower amounts in C20 fatty acids, with monoenoic C20 forms not present. This pattern likely separates primary consumers (fungivores) from predators (nematode feeders). The isotopic discrimination in 13C for bulk Collembola ranged between −2.6 and 1.4‰ and was dependent on fungal species and C 3/C 4 system, suggesting differences at metabolic branch points and/or isotope discrimination of enzymes. Comparison of δ 13C values in individual fatty acids between consumer and diet generally showed depletion (i.e. de novo synthesis) or no changes (i.e. dietary routing), but the fractionation was not uniform and affected by the type of ingested food. Fatty acid carbon isotopes were more variable than those of bulk tissues, likely due to both the distrimination by enzymes and the different lipid origin (i.e. neutral or polar fraction).

15N stable isotope labelling of slugs (Gastropoda: Pulmonata)

SummaryStable isotope tracers are a promising tool for investigating the ecology of terrestrial slugs, including predator‐prey relationships, migration behaviour, nutrient turnover and dietary routing. The objective of the present feasibility study was to label two economically important slug groups, Deroceras reticulatum and keeled slugs (families Limacidae and Milacidae, respectively), with the stable isotope 15N under controlled laboratory conditions. Significant isotopic enrichment in slug tissue was detected after 4 days and persisted for at least 10 days after slugs had been fed on 15N enriched food for a period of 15 days. The time course of 15N uptake into slug tissues and its relation to food consumption were well described mathematically. Estimated mean 15N assimilation efficiencies from labelled maize mixed with unlabelled wheat bran were 30% and 38%, respectively, for the species groups studied. These findings suggest that slugs can be readily and efficiently labelled and that it is feasible to devise protocols for producing large numbers of isotopically labelled slugs for use in ecological studies. A simple method is described for the collection and analysis of cutaneous mucus from individual slugs which can be used to test uniformity of isotopic labelling.