Compound‐specific stable isotope analysis (
Compound‐specific stable isotope analysis of individual amino acids (CSIA‐AA) has emerged as a transformative approach to estimate consumer trophic positions (TPCSIA) that are internally indexed to primary producer nitrogen isotope baselines. Central to accurate TPCSIAestimation is an understanding of beta ( This meta‐analysis fulfils a pressing need to comprehensively evaluate relevant sources of We show that variation in Our results highlight that primary producer
- NSF-PAR ID:
- 10377312
- Publisher / Repository:
- Wiley-Blackwell
- Date Published:
- Journal Name:
- Methods in Ecology and Evolution
- Volume:
- 12
- Issue:
- 10
- ISSN:
- 2041-210X
- Format(s):
- Medium: X Size: p. 1750-1767
- Size(s):
- ["p. 1750-1767"]
- Sponsoring Org:
- National Science Foundation
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Abstract CSIA ) of amino acids (AA ) has rapidly become a powerful tool in studies of food web architecture, resource use, and biogeochemical cycling. However, applications to avian ecology have been limited because no controlled studies have examined the patterns inAA isotope fractionation in birds. We conducted a controlledCSIA feeding experiment on an avian species, the gentoo penguin (Pygoscelis papua ), to examine patterns in individualAA carbon and nitrogen stable isotope fractionation between diet (D) and consumer (C) (Δ13CC‐Dand Δ15NC‐D, respectively). We found that essentialAA δ 13C values and sourceAA δ 15N values in feathers showed minimal trophic fractionation between diet and consumer, providing independent but complimentary archival proxies for primary producers and nitrogen sources respectively, at the base of food webs supporting penguins. Variations in nonessentialAA Δ13CC‐Dvalues reflected differences in macromolecule sources used for biosynthesis (e.g., protein vs. lipids) and provided a metric to assess resource utilization. The avian‐specific nitrogen trophic discrimination factor (TDF Glu‐Phe= 3.5 ± 0.4‰) that we calculated from the difference in trophic fractionation (Δ15NC ‐D) of glutamic acid and phenylalanine was significantly lower than the conventional literature value of 7.6‰. Trophic positions of five species of wild penguins calculated using a multi‐TDFG lu‐Pheequation with the avian‐specificTDFG lu‐Phevalue from our experiment provided estimates that were more ecologically realistic than estimates using a singleTDFG lu‐Pheof 7.6‰ from the previous literature. Our results provide a quantitative, mechanistic framework for the use ofCSIA in nonlethal, archival feathers to study the movement and foraging ecology of avian consumers. -
Rationale Ecologists increasingly determine the δ15N values of amino acids (AA) in animal tissue; “source” AA typically exhibit minor variation between diet and consumer, while “trophic” AA have increased δ15N values in consumers. Thus, trophic‐source δ15N offsets (i.e., Δ15NT‐S) reflect trophic position in a food web. However, even minor variations in δ15Nsource AAvalues may influence the magnitude of offset that represents a trophic step, known as the trophic discrimination factor (i.e., TDFT‐S). Diet digestibility and protein content can influence the δ15N values of bulk animal tissue, but the effects of these factors on AA Δ15NT‐Sand TDFT‐Sin mammals are unknown.
Methods We fed captive mice (
) either (A) a low‐fat, high‐fiber diet with low, intermediate, or high protein; or (B) a high‐fat, low‐fiber diet with low or intermediate protein. Mouse muscle and dietary protein were analyzed for bulk tissue δ15N using elemental analyzer‐isotope ratio mass spectrometry (EA‐IRMS), and were also hydrolyzed into free AA that were analyzed for δ15N using gas chromatography‐combustion‐IRMS.Mus musculus Results As dietary protein increased, Δ15NConsumer‐Dietslightly declined for bulk muscle tissue in both experiments; increased for AA in the low‐fat, high‐fiber diet (A); and remained the same or decreased for AA in the high‐fat, low‐fiber diet (B). The effects of dietary protein on Δ15NT‐Sand on TDFT‐Svaried by AA but were consistent between variables.
Conclusions Diets were less digestible and included more protein in Experiment A than in Experiment B. As a result, the mice in Experiment A probably oxidized more AA, resulting in greater Δ15NConsumer‐Dietvalues. However, the similar responses of Δ15NT‐Sand of TDFT‐Sto diet variation suggest that if diet samples are available, Δ15NT‐Saccurately tracks trophic position. If diet samples are not available, the patterns presented here provide a basis to interpret Δ15NT‐Svalues. The trophic‐source offset of Pro‐Lys did not vary across diets, and therefore may be more reliable for omnivores than other offsets (e.g., Glu‐Phe).
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null (Ed.)We investigated the response of an open-ocean plankton food web to a major ecosystem perturbation event, the Hawaiian lee cyclonic eddy Opal, using compound-specific isotopic analyses of amino acids (CSIA-AA) of individual zooplankton taxa. We hypothesized that the massive diatom bloom that characterized Opal would lead to a shorter food chain. Using CSIA-AA, we differentiated trophic position (TP) changes that arose from altered transfers through protistan microzooplankton, versus metazoan carnivory, and assessed the variability at the base of the food web. Contrary to expectation, zooplankton TPs were higher in the eddy than in ambient control waters (up to 0.8 trophic level), particularly for suspension feeders close to the food-web base. Most of the effect was due to increased trophic transfers through protistan consumers, indicating a general shift up, not down, of grazing and remineralization in the microbial food web. Eucalanus sp., which was 15-fold more abundant inside compared to outside of the eddy, was the only taxon observed to be a true herbivore (TP = 2.0), consistent with a high phenylalanine (Phe) δ 15 N value indicating feeding on nitrate-fueled diatoms in the lower euphotic zone. Oncaea sp., an aggregate-associated copepod, had the largest (1.5) TP difference, and lowest Phe δ 15 N, suggesting that detrital particles were local hot spots of enhanced microbial activity. Rapid growth rates and trophic flexibility of protistan microzooplankton apparently allow the microbial community to reorganize to bloom perturbations, as microzooplankton remain the primary phytoplankton grazers—despite the dominance of large diatoms—and are heavily preyed on by the mesozooplankton.more » « less
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Abstract Sponges are a crucial component of Caribbean coral reef ecosystem structure and function. In the Caribbean, many sponges show a predictable increase in percent cover or abundance as depth increases from shallow (< 30 m) to mesophotic (30–150 m) depths. Given that sponge abundances are predicted to increase in the Caribbean as coral cover declines, understanding ecological factors that control their distribution is critical. Here we assess if sponge cover increases as depth increases into the mesophotic zone for three common Caribbean reef sponges,
Xestospongia muta ,Agelas tubulata , andPlakortis angulospiculatus , and use stable isotope analyses to determine whether shifts in trophic resource utilization along a shallow to mesophotic gradient occurred. Ecological surveys show that all target sponges significantly increase in percent cover as depth increases. Using bulk stable isotope analysis, we show that as depth increases there are increases in the δ13C and δ15N values, reflecting that all sponges consumed more heterotrophic picoplankton, with low C:N ratios in the mesophotic zone. However, compound‐specific isotope analysis of amino acids (CSIA‐AA) shows that there are species‐specific increases in δ13CAAand δ15NAAvalues.Xestospongia muta andP. angulospiculatus showed a reduced reliance on photoautotrophic resources as depth increased, whileA. tubulata appears to rely on heterotrophy at all depths. The δ13CAAand δ15NAAvalues of these sponges also reflect species‐specific patterns of host utilization of both POM and dissolved organic matter (DOM), its subsequent re‐synthesis, and translocation, by their microbiomes. -
Rationale Nitrogen isotopic compositions (δ15N) of source and trophic amino acids (AAs) are crucial tracers of N sources and trophic enrichments in diverse fields, including archeology, astrobiochemistry, ecology, oceanography, and paleo‐sciences. The current analytical technique using gas chromatography‐combustion‐isotope ratio mass spectrometry (GC/C/IRMS) requires derivatization, which is not compatible with some key AAs. Another approach using high‐performance liquid chromatography‐elemental analyzer‐IRMS (HPLC/EA/IRMS) may experience coelution issues with other compounds in certain types of samples, and the highly sensitive nano‐EA/IRMS instrumentations are not widely available.
Methods We present a method for high‐precision δ15N measurements of AAs (δ15N‐AA) optimized for canonical source AA‐phenylalanine (Phe) and trophic AA‐glutamic acid (Glu). This offline approach entails purification and separation via high‐pressure ion‐exchange chromatography (IC) with automated fraction collection, the sequential chemical conversion of AA to nitrite and then to nitrous oxide (N2O), and the final determination of δ15N of the produced N2O via purge‐and‐trap continuous‐flow isotope ratio mass spectrometry (PT/CF/IRMS).
Results The cross‐plots of δ15N of Glu and Phe standards (four different natural‐abundance levels) generated by this method and their accepted values have a linear regression slope of 1 and small intercepts demonstrating high accuracy. The precisions were 0.36‰–0.67‰ for Phe standards and 0.27‰–0.35‰ for Glu standards. Our method and the GC/C/IRMS approach produced equivalent δ15N values for two lab standards (McCarthy Lab AA mixture and cyanobacteria) within error. We further tested our method on a wide range of natural sample matrices and obtained reasonable results.
Conclusions Our method provides a reliable alternative to the current methods for δ15N‐AA measurement as IC or HPLC‐based techniques that can collect underivatized AAs are widely available. Our chemical approach that converts AA to N2O can be easily implemented in laboratories currently analyzing δ15N of N2O using PT/CF/IRMS. This method will help promote the use of δ15N‐AA in important studies of N cycling and trophic ecology in a wide range of research areas.