Compound-specific Nitrogen Isotope Research Articles

Trophic Transfer of Halogenated Organic Pollutants in a Wetland Food Web: Insights from Compound-Specific Nitrogen Isotope of Amino Acids and Food Source Analysis.

A trophic position (TP) model (TPmix model) that simultaneously considered trophic discrimination factor and βGlu/Phe variations was developed in this study and was first applied to investigate the trophic transfer of halogenated organic pollutants (HOPs) in wetland food webs. The TPmix model characterized the structure of the wetland food web more accurately and significantly improved the reliability of TMF compared to the TPbulk, TPAAs, and TPsimmr models, which were calculated based on the methods of stable nitrogen isotope analysis of bulk, traditional AAs-N-CSIA, and weighted βGlu/Phe, respectively. Food source analysis revealed three interlocking food webs (kingfisher, crab, and frogs) in this wetland. The highest HOP biomagnification capacities (TMFmix) were found in the kingfisher food web (0.24-82.0), followed by the frog (0.08-34.0) and crab (0.56-11.7) food webs. The parabolic trends of TMFmix across combinations of log KOW in the frog food web were distinct from those of aquatic food webs (kingfisher and crab), which may be related to differences in food web composition and HOP bioaccumulation behaviors between aquatic and terrestrial organisms. This study provides a new tool to accurately study the trophic transfer of contaminants in wetlands and terrestrial food webs with diverse species and complex feeding relationships.

Deconvolving mechanisms of particle flux attenuation using nitrogen isotope analyses of amino acids

AbstractParticulate organic matter settling out of the euphotic zone is a major sink for atmospheric carbon dioxide and serves as a primary food source to mesopelagic food webs. Degradation of this organic matter encompasses a suite of mechanisms that attenuate flux, including heterotrophic metabolic processes of microbes and metazoans. The relative contributions of microbial and metazoan heterotrophy to flux attenuation, however, have been difficult to determine. We present results of compound specific nitrogen isotope analysis of amino acids of sinking particles from sediment traps and size‐fractionated particles from in situ filtration between the surface and 500 m at Ocean Station Papa, collected during NASA EXPORTS (EXport Processes in the Ocean from RemoTe Sensing). With increasing depth, we observe: (1) that, based on the δ15N values of threonine, fecal pellets dominate the sinking particle flux and that attenuation of downward particle flux occurs largely via disaggregation in the upper mesopelagic; (2) an increasing trophic position of particles in the upper water column, reflecting increasing heterotrophic contributions to the nitrogen pool and the loss of particles via remineralization; and (3) increasing δ15N values of source amino acids in submicron and small (1–6 μm) particles, reflecting microbial particle solubilization. We further employ a Bayesian mixing model to estimate the relative proportions of fecal pellets, phytodetritus, and microbially degraded material in particles and compare these results and our interpretations of flux attenuation mechanisms to other, independent methods used during EXPORTS.

Compound-specific Nitrogen Isotope and Radiocarbon Analyses of Amino Acids in Biological Samples

Compound-specific Nitrogen Isotope and Radiocarbon Analyses of Amino Acids in Biological Samples

Pig management in the Neolithic Near East and East Asia clarified with isotope analyses of bulk collagen and amino acids

The chemical analysis of animal bones from ancient sites has become a common approach in archeological research investigating animal utilization and domestication by past humans. Although several chemical indicators have been used to determine pig management practices in ancient societies, one indicator that can clarify human-animal relationships in the early stages of domestication is the change in the animal’s diet from its wild diet, which can be detected using isotope analysis of its bones. Omnivores, such as boars, are assumed to have shared foods with humans as their interaction increased, and a shift in the isotopic (carbon and nitrogen) compositions of their bone collagen toward humans are considered evidence of domestication. This approach has found evidence of early-stage pig management with human leftovers and feces in prehistoric East Asia, including in Neolithic China, Korea and Japan. However, in the Near East, one of the origins of animal domestication, even individual animals considered to be domesticated pigs according to zooarcheological data (such as morphological characteristics and mortality patterns) display isotopic compositions of bulk collagen that differ from those of humans but are close to those of herbivores. This result indicates that these pigs were fed special foods, such as legumes, rather than human leftovers or feces. However, the carbon and nitrogen isotopic compositions of the bulk collagen of herbivores found at the same sites showed huge variations, so the interpretation of the pigs’ diet is consequently unclear. In this study, a compound-specific nitrogen isotope analysis was used to clarify the pig diet and management strategies unique to the Neolithic Near East, Turkey and Syria, together with a carbon and nitrogen isotope analysis of bulk collagen. This study examines the diversity of pig management techniques in early agricultural societies and their relationship with the availability of other domestic animals and farming practices.

Compound specific isotope evidence points to use of freshwater resources as weaning food in Middle Neolithic Paris Basin

ObjectivesA clear understanding of past weaning practices can provide invaluable insights into social issues such as infant care, fertility rate, and demographic patterns in past societies. This study presents the first archeological research employing compound specific isotope analysis (CSIA) for the reconstruction of past weaning practices.MethodsWeaning practices of two Middle Neolithic communities in the Paris Basin region: Balloy (BLR) and Vignely (VPB), are evaluated by combining previously published bone collagen stable carbon, nitrogen, and sulfur (n = 66) isotope analysis with new compound specific carbon and nitrogen isotope compositions of bone collagen (n = 10).ResultsOur results demonstrate that the diets of individuals from BLR and VPB likely incorporated freshwater resources. The signals of freshwater resources consumption are even stronger among subadults, suggesting that freshwater resources were used as weaning food at these sites.ConclusionsThe implications of our result are threefold. Currently many CSIA studies in archeology only involve either carbon or nitrogen. Our data shows that it is important to conduct CSIA on both carbon and nitrogen for a more integrated picture. Secondly, our data demonstrates that the use of a protein‐based weaning food—instead of a starch‐based weaning food (such as cereal gruel)—was likely more prevalent among the Middle Neolithic communities in the Paris Basin Region than previously thought. The finding thus prompts a rethinking of the role of protein‐based weaning food in other archeological contexts. Lastly, the common assumption that weaning foods and adult diets share similar isotopic compositions can be problematic, as the use of protein‐based, high trophic‐level weaning foods can skew the δ 15N weaning curve and produce an erroneously late estimation for weaning ages.

Isotopic insights into the early Medieval (600–1100 CE) diet in the Luistari cemetery at Eura, Finland

In this article, we present the results of an isotopic study of diet for the early medieval (Merovingian, Viking, Early Christian) humans buried in the unique Luistari cemetery at Eura (ca. 600–1400 CE), southwestern Finland, the largest cemetery of the region. Isotope analysis was conducted on 37 humans for dentine and bone collagen (δ13C, δ15N, and δ34S), and five of them were also studied using compound-specific nitrogen isotope analysis. Dental enamel and/or bone carbonate δ13C values were studied from altogether 65 humans, five cattle, and five sheep/goats. The bone and dentine collagen and carbonate data show that throughout the centuries, freshwater fish was a stable part of the diet for the population. Our results do not show systematic dietary differences between estimated males and females, but differences can be large on the individual level. We also discovered a possible temporal change in the enamel carbonate ẟ13C values that could be related to the increasing role of carbohydrates (e.g., crops) in the diet. Luistari burials are well comparable to contemporary Swedish Viking trading communities like Birka in their higher protein intake. But contrary to the wider Viking network, they do not show the same marine signal.

Environmental fate and trophic transfer of synthetic musk compounds and siloxanes in Geum River, Korea: Compound-specific nitrogen isotope analysis of amino acids for accurate trophic position estimation.

Despite the extensive usage of synthetic musk compounds (SMCs) and siloxanes in various personal care products (PCPs), trophic magnification of such chemicals in aquatic environments remains unexplored. In June and September 2020, eleven SMCs and nineteen siloxanes were measured in water, sediments, and biota. Samples were collected from two sites where levels were expected to be influenced by the distance from the wastewater treatment plant (WWTP) in the Geum River, Republic of Korea, were expected. High concentrations of SMCs and siloxanes entered through WWTP were measured in water, sediment, and biota at the both sites and both seasons. The δ15N of amino acids provided a high-resolution food web and accurate trophic position (TP), which is an important factor for determining the trophic magnification factor (TMF). Among 24 TMFs, 19 of them were <1, ranging 0.7-0.8 for 1,3,4,6,7,8‑hexahydro‑4,6,6,7,8,8‑hexamethyl‑cyclopenta‑γ‑2‑benzopyran (HHCB), 0.6-0.8 for 6-Acetyl-1,1,2,4,4,7-hexamethyltetralin (AHTN), 0.7-0.8 for 4-tert-Butyl-3,5-dinitro-2,6-dimethylacetophenone (MK), 0.7-0.9 for octamethylcyclotetrasiloxane (D4), 0.1-0.4 for decamethylcyclopentasiloxane (D5), and 0.04-0.8 for dodecamethylcyclohexasiloxane (D6), and the remaining ones including HHCB, AHTN, MK, and D4 showed values close to 1 or slightly higher (TMF range: 1.0-2.3) indicating no or a little trophic magnification. The TMFs of these compounds were constant across sites and seasons. The TMF values of PCPs might be affected by species specificity and food web structure rather than by chemical properties such as log Kow, which describes a wide range of TMF values in various environments. This study presents valuable implications for assessing risk and managing environmental fate and trophic transfer of SMCs and siloxanes in freshwater environments.

Compound-Specific Nitrogen Isotope Analysis of Amino Acids in Eye Lenses as a New Tool to Reconstruct the Geographic and Trophic Histories of Fish

Fish migration has always played an essential role in marine conservation and fisheries management. However, migration patterns are changing globally alongside changing ocean conditions. This affects the spatial scale of required governance and, consequently, our food supply. Technological advances in the bio-tracer approach using isotopes would make it possible to monitor fish migration differently from the conventional bio-logging method. Eye lenses of fish are incrementally grown and metabolically inert. Therefore, stable isotope analysis of eye lenses can reconstruct the geographic and trophic histories of fish. However, it is difficult to distinguish between the two pieces of information: migration and changing trophic relationships with the conventional bulk stable isotope approach. In this study, we measured the nitrogen isotope ratios of individual amino acids in eye lenses of chub mackerel (n = 3) from the western North Pacific to test whether the two pieces of information could be distinguished. A maximum of 34 growth layers was obtained from a single eye lens. For assigning the isotopic trends observed in lens tissues to specific life stages or size classes, we established a relationship between lens size and fork length (FL) as a reference framework. The reconstructed nitrogen isotope chorology of chub mackerel showed an increase in trophic position of about 1 unit from juvenile (FL ∼30 mm) to adult stages (FL ∼300 mm). The variation in nitrogen isotope ratios of phenylalanine (a source amino acid) in the eye lenses was –3 to 5‰. This baseline variability could be used for estimating fish movements. This study proposed compound-specific nitrogen isotope analysis of amino acids in eye lenses as a new tool capable of reconstructing the geographic and trophic histories of migratory fish.

Trophic niche of seabirds on the Barton Peninsula, King George Island, Antarctica

Drastic sea ice retreats in the Antarctic Peninsula, and the consequent environmental changes have brought about the consequences of biological adaptation and food competition. The isotopic niche is reflected by the ecological position and functional role of a species, which can be altered depending on these environmental changes. We assessed the isotopic niche and trophic positions (TP) of 3 seabirds on the Barton Peninsula, King George Island, Antarctica, to understand ecological interaction among the species. The average TP of Antarctic krill (Euphausia superba) (hereafter “krill”) (2.6 ± 0.1) was estimated by using the compiled published data of the compound-specific nitrogen isotope ratio of glutamic acid and phenylalanine in the Southern Ocean, and TP values of the seabirds based on that of krill to be reliable according to ecological knowledge. Our results on the overlap of the isotopic niches of seabirds suggests potential diet competition among the consumers, such as the brown skua (Stercorarius antarcticus) and south polar skua (Stercorarius maccormicki), whereas the distinct and broad isotopic niche width of kelp gulls (Larus dominicanus) suggesting that their adaptation through generalization was related to their extended habitat and various food sources. Our research indicates that isotopic niche of seabirds reflects their survival strategy for food competition. Consequently, these seabirds can be easily influenced by alternative food sources, including terrestrial and human-derived sources, by rapid environmental changes, indicating that they are valuable as key environmental species and require long-term monitoring in the Antarctic Peninsula.

Nitrogen isotope fractionation of amino acids from a controlled study on the green turtle (Chelonia mydas): expanding beyond Glx/Phe for trophic position

Compound specific nitrogen isotope analysis of amino acids (AA CSIA) has been used to study the trophic ecology of many marine organisms because of its ability to characterize the δ15N value at the base of a food web while simultaneously providing trophic position information from a single tissue sample of a consumer. Although application of this method is becoming more widely used in trophic studies, critical assumptions such as amino acid designation and trophic discrimination factors remain unresolved for many taxa and tissue types. In this study, we used captive green turtles (Chelonia mydas) in a controlled feeding study to determine and evaluate individual amino acid isotope fractionation patterns and trophic discrimination factors (TDFs) for skin and plasma. Our data build on findings from previous studies that show that the traditional paradigm of using glutamic acid and phenylalanine as trophic and source amino acids, respectively, for trophic position estimation is consistent for green turtles; we also provide evidence to support the potential utility of other combinations of amino acids for trophic position calculations. Green turtle TDFs for glutamic acid and phenylalanine were 4.0 ± 0.5 ‰ and 6.6 ± 0.7 ‰ for skin and plasma, respectively. We found serine to have the largest Δδ15N (turtle-diet discrimination) value of all amino acids in both skin and plasma, while lysine Δδ15N values were consistent for use as a source amino acid in skin and plasma tissue. We evaluated our TDFs by estimating trophic position of four sea turtle species using previously published amino acid isotope data. These results indicate that for green turtles, the traditional use of glutamic acid and phenylalanine along with a combination of trophic amino acids paired with lysine can provide robust trophic position estimates. Moreover, we highlight the taxa and tissue-specific nature of amino acid nitrogen isotope fractionation patterns in green turtle tissues and underscore that well-approximated TDFs can refine trophic position estimates for sea turtles.

Variability of trophic magnification factors as an effect of estimated trophic position: Application of compound-specific nitrogen isotope analysis of amino acids

The trophic magnification of persistent organic pollutants (POPs), which is the relationship between POP concentration and the trophic position (TPs) of an organism, is considered an important factor for prioritizing chemicals of concern in the environment. Organismal TPs are typically based on nitrogen isotope ratios of bulk tissue (δ15Nbulk). In this study, nitrogen isotope ratios of amino acids (δ15NAAs), a more precise approach for TP estimation (TPAAs), was applied and compared with estimations of TP based on δ15Nbulk (TPbulk) in marine organisms living in Masan Bay, South Korea. Compound-specific isotope analysis of the amino acids (CSIA-AAs) in fish samples allows us to calculate robust TPs by correcting the variation in baseline isotope values with use of the δ15Nbulk technique. In a benthic food chain, this approach reveals more significant magnification trends for POPs [polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and organochlorine pesticides (OCPs)] than those revealed by analysis of the relationship between TPbulk and POPs. The trophic magnification factors (TMF) associated with TPAAs were significant for some POPs, especially pp′-DDD and chlordane. The results presented in this study suggest that TP calculations based on δ15NAAs are an effective tool for characterizing trophic magnification trends related to the fates of various pollutants.

Paired bulk organic and individual amino acid δ15N analyses of bivalve shell periostracum: A paleoceanographic proxy for water source variability and nitrogen cycling processes

Abstract Developing high-resolution, well-dated, marine proxies of environmental, climatic, and oceanographic conditions is critical in order to advance our understanding of the ocean’s role in the global climate system. While some work has investigated bulk and compound specific stable nitrogen isotope (δ15N) values in bivalve shells as proxies for environmental variability, the small concentrations of nitrogen found in the organic matrix of the shell calcium carbonate (hereafter carbonate) makes developing high resolution records challenging. This study investigates the potential of using the bulk and amino acid δ15N values of bivalve periostracum, the protein layer on the outside of the shell, as a proxy archive of nitrogen cycling processes and water source variability. Bulk δ15N values were measured on the periostracum, carbonate, and adductor muscle of Arctica islandica shells collected in the Gulf of Maine. Increased variability of isotopic values across growth lines compared to along growth lines support mechanistic reasoning based on growth processes that periostracum is recording changes in δ15N values over the course of the clam’s lifetime (up to 500 years). In addition, the statistically significant relationship between periostracum δ15N values and contemporaneous carbonate δ15N values of the same shell (r = 0.82, p  Compound specific δ15N analyses of the periostracum of A. islandica shells were used to determine that the calculated trophic position of the clams in this study (1.4 ± 0.4) did not change significantly between 1783 and 1997. Phenylalanine δ15N values over the last 70 years show similar trends to that of the bulk record, suggesting that changes in bulk δ15N values over that time period are related to changes in baseline δ15N values. Periostracum δ15N values from shells collected in the western Gulf of Maine have decreased by ∼1‰ since the mid-1920s. This trend (−0.008‰/year) is not statistically different from the trend of previously published δ15N values of deep-sea corals from the entrance to the Gulf of Maine over the same time period. This coral record has been shown to indicate a shift in water source in the region and therefore the similarity between the two records suggest that changes in periostracum δ15N values are reflecting broader North Atlantic hydrographic changes. Our study introduces a new, high-resolution, and absolutely dated paleoceanographic proxy of baseline δ15N values, presenting the opportunity for future reconstructions of aspects of nitrogen cycling and water source changes in the global oceans.

Comparison of trophic magnification factors of PCBs and PBDEs in Tokyo Bay based on nitrogen isotope ratios in bulk nitrogen and amino acids

Trophic positions (TPs) are key parameters for determining trophic magnification factors (TMFs) of environmental pollutants. We used compound-specific nitrogen isotope analysis of amino acids (CSIA-AA), with which we expected to obtain precise TPs, to estimate the TMFs of polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) in the pelagic and benthic food webs of Tokyo Bay. The objective of the present study was to evaluate the difference between TMFs derived from TPs estimated by CSIA-AA (TPGlu/Phe) and TMFs derived from TPs estimated by bulk stable isotope analysis (TPbulk). TPGlu/Phe tended to be lower than TPbulk for many pelagic and benthic fishes. In the pelagic food web, the slopes for the relationships between PCB concentrations and TPGlu/Phe exceeded those between PCB concentrations and TPbulk for many congeners in samples from December 2014, and TMFGlu/Phe values (0.89–6.9) were higher than TMFbulk values (0.92–3.8). However, there were no statistically significant differences between the slopes of the relationships between PCB and PBDE concentrations and TPbulk or TPGlu/Phe for any of the congeners (analysis of covariance, P ≥ 0.08). In the benthic food web, PCB concentrations correlated with TPGlu/Phe for 9 PCB congeners in the samples collected during December 2014, although the concentrations of none of the PCB congeners correlated with TPbulk. A decrease of TMFGlu/Phe and TMFbulk values from December 2014 to May 2015 suggested that variation of PCB and PBDE concentrations in the organisms had more effect than TPs on the TMFs.

Long-term trends in the foraging ecology and habitat use of an endangered species: an isotopic perspective.

Evaluating long-term drivers of foraging ecology and population productivity is crucial for providing ecological baselines and forecasting species responses to future environmental conditions. Here, we examine the trophic ecology and habitat use of North Atlantic leatherback turtles (St. Croix nesting population) and investigate the effects of large-scale oceanographic conditions on leatherback foraging dynamics. We used bulk and compound-specific nitrogen isotope analysis of amino acids (CSIA-AA) to estimate leatherback trophic position (TP) over an 18-year period, compare these estimates with TP estimates from a Pacific leatherback population, and elucidate the pre-nesting habitat use patterns of leatherbacks. Our secondary objective was to use oceanographic indices and nesting information from St. Croix leatherbacks to evaluate relationships between trophic ecology, nesting parameters, and regional environmental conditions measured by the North Atlantic Oscillation (NAO) and Atlantic Multidecadal Oscillation. We found no change in leatherback TP over time and no difference in TP between Atlantic and Pacific leatherbacks, indicating that differences in trophic ecology between populations are an unlikely driver of the population dichotomy between Pacific and Atlantic leatherbacks. Isotope data suggested that St. Croix leatherbacks inhabit multiple oceanic regions prior to nesting, although, like their conspecifics in the Pacific, individuals exhibit fidelity to specific foraging regions. Leatherback nesting parameters were weakly related to the NAO, which may suggest that positive NAO phases benefit St. Croix leatherbacks, potentially through increases in resource availability in their foraging areas. Our data contribute to the understanding of leatherback turtle ecology and potential mechanistic drivers of the dichotomy between populations of this protected species.

Use of compound-specific nitrogen isotope analysis of amino acids in trophic ecology: assumptions, applications, and implications

Knowledge of the diet source and trophic position of organisms is fundamental in food web science. Since the 1980s, stable isotopes of light elements such as 13C and 15N have provided unique information on the food web structure in a variety of ecosystems. More recently, novel isotope tools such as stable isotopes of heavy elements, radioisotopes, and compound-specific isotope analysis, have been examined by researchers. Here I reviewed the use of compound-specific nitrogen isotope analysis of amino acids (CSIA-AA) as a useful dietary tracer in food web ecology. Its three key features—(1) offsetting against isotopic variation; (2) universality of the trophic discrimination factor; and (3) sensitivity to source mixing—were compared with conventional isotope analysis for the bulk tissue of organisms. These three advantages of CSIA-AA will allow future researchers to (1) estimate the integrated trophic position (iTP) of animal communities; (2) infer trophic transfer efficiency (TTE) in food webs; and (3) quantify contributions from different resources to animals, all of which are crucial for understanding the relationship between biodiversity and multi-trophic ecosystem functioning. Further development of trophic ecology will be facilitated by both methodological refinement of CSIA-AA and its application to a wider range of organisms, food webs, and ecosystems.

Amino Acid Specific Stable Nitrogen Isotope Values in Avian Tissues: Insights from Captive American Kestrels and Wild Herring Gulls.

Through laboratory and field studies, the utility of amino acid compound-specific nitrogen isotope analysis (AA-CSIA) in avian studies is investigated. Captive American kestrels (Falco sparverius) were fed an isotopically characterized diet and patterns in δ15N values of amino acids (AAs) were compared to those in their tissues (muscle and red blood cells) and food. Based upon nitrogen isotope discrimination between diet and kestrel tissues, AAs could mostly be categorized as source AAs (retaining baseline δ15N values) and trophic AAs (showing 15N enrichment). Trophic discrimination factors based upon the source (phenylalanine, Phe) and trophic (glutamic acid, Glu) AAs were 4.1 (muscle) and 5.4 (red blood cells), lower than those reported for metazoan invertebrates. In a field study involving omnivorous herring gulls (Larus argentatus smithsonianus), egg AA isotopic patterns largely retained those observed in the laying female's tissues (muscle, red blood cells, and liver). Realistic estimates of gull trophic position were obtained using bird Glu and Phe δ15N values combined with β values (difference in Glu and Phe δ15N in primary producers) for aquatic and terrestrial food webs. Egg fatty acids were used to weight β values for proportions of aquatic and terrestrial food in gull diets. This novel approach can be applied to generalist species that feed across ecosystem boundaries.

A reliable compound-specific nitrogen isotope analysis of amino acids by GC-C-IRMS following derivatisation into N-pivaloyl-iso-propyl (NPIP)esters for high-resolution food webs estimation.

The signatures of natural stable nitrogen isotopic composition (δ(15)N) of individual amino acid (AA) have been confirmed to be a potentially effective tool for elucidating nitrogen cycling and trophic position of various organisms in food webs. In the present study, a two-stage derivatisation approach of esterification followed by acylation was evaluated. The biological samples underwent acid hydrolysis and the released individual AA was derivatived into corresponding N-pivaloyl-isopropyl (NPIP) esters for nitrogen isotopic analysis in gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). Usually, 13 individual AA derivatives were separated with fine baseline resolution based on a nonpolar gas chromatography column (DB-5ms). The minimum sample amount required under the presented conditions is larger than 20ngN on column in order to accurately determine the δ(15)N values. The δ(15)N values determined by GC-C-IRMS with a precision of better than 1‰, were within 1‰ after empirical correction compared to the corresponding measured by element analysis (EA)-IRMS. Bland-Altman plot showed highly consistency of the δ(15)N values determined by the two measurement techniques. Cation-exchange chromatography was applied to remove interfering fraction from the extracts of plant and animal samples and without nitrogen isotope fractionation during the treatment procedure. Moreover, this approach was carried out to estimate the trophic level of various natural organisms in a natural lake environment. Results highly proved that the trophic level estimated via the presented AA method well reflected the actual food web structure in natural environments.

Compound-Specific Isotope Analysis of Amino Acid Labeling with Stable Isotope Nitrogen (15N) in Higher Plants

Individual free amino acid δ15N values in plant tissue reflect the metabolic pathways involved in their biosynthesis and catabolism and could thus aid understanding of environmental stress and anthropogenic effects on plant metabolism. In this study, compound-specific nitrogen isotope analysis of amino acid by gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) was carried out to determine individual free amino acid δ15N values. High correlations were observed between the δ15N values obtained by GC-C-IRMS and elemental analyzer-isotope ratio mass spectrometry (EA-IRMS) determinations, and the mean precision measured was better than 1 ‰. Cation-exchange chromatography was employed to purify the sample, and the difference between prior to and following passage through the resin was within 1 ‰. The amino acid δ15N values of plant leave samples following incubation in 15N-nitrate at different time points were determined. A typical foliar free amino acid 15N-enrichment pattern was found, and glutamine was the most rapidly labeled amino acid; other amino acids derived from the GS-GOGAT cycle were also enriched. The pyruvate family amino acids were labeled less quickly followed by the aromatic amino acids. This study highlighted that amino acid metabolism pathways had a major effect on the δ15N values. With the known amino acid metabolism pathways and δ15N values determined by the presented method, the influence of various external factors on the metabolic cycling of amino acid can be understood well.

Amino acid compositions in heated carbonaceous chondrites and their compound-specific nitrogen isotopic ratios

A novel method has been developed for compound-specific nitrogen isotope compositions with an achiral column which was previously shown to offer high precision for nitrogen isotopic analysis. We applied the method to determine the amino acid contents and stable nitrogen isotopic compositions of individual amino acids from the thermally metamorphosed (above 500 °C) Antarctic carbonaceous chondrites Ivuna-like (CI)1 (or CI-like) Yamato (Y) 980115 and Ornans-like (CO)3.5 Allan Hills (ALH) A77003 with the use of gas chromatography/combustion/isotope ratio mass spectrometry. ALHA77003 was deprived of amino acids due to its extended thermal alteration history. Amino acids were unambiguously identified in Y-980115, and the δ15N values of selected amino acids (glycine +144.8 ‰; α-alanine +121.2 ‰) are clearly extraterrestrial. Y-980115 has experienced an extended period of aqueous alteration as indicated by the presence of hydrous mineral phases. It has also been exposed to at least one post-hydration short-lived thermal metamorphism. Glycine and alanine were possibly produced shortly after the accretion event of the asteroid parent body during the course of an extensive aqueous alteration event and have abstained from the short-term post-aqueous alteration heating due to the heterogeneity of the parent body composition and porosity. These carbonaceous chondrite samples are good analogs that offer important insights into the target asteroid Ryugu of the Hayabusa-2 mission, which is a C-type asteroid likely composed of heterogeneous materials including hydrated and dehydrated minerals.

Re-evaluation of nutrient sources for deep-sea wood-boring bivalves using the isotopic composition of bulk C, N, S, and amino acid nitrogen

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 540:157-165 (2015) - DOI: https://doi.org/10.3354/meps11510 Re-evaluation of nutrient sources for deep-sea wood-boring bivalves using the isotopic composition of bulk C, N, S, and amino acid nitrogen Toshiro Yamanaka1,*, Sho Shimamura1,4, Yoshito Chikaraishi2, Takuma Haga3, Yoshihiro Fujiwara2 1Graduate school of Natural Science and Technology, Okayama University, 1-1 Naka 3-chome, Tsushima, Kita-ku, Okayama 700-8530, Japan 2Japan Agency for Marine-Earth Science and Technology, 2-15 Natsushima-cho, Yokosuka 237-0061, Japan 3Toyohashi Museum of Natural History, 1-238 Oana, Oiwa, Toyohashi 441-3147, Japan 4Present address: Japan Meteorological Agency, 1-3-4 Otemachi, Chiyoda-ku, Tokyo 100-8122, Japan *Corresponding author: toshiroy@cc.okayama-u.ac.jp ABSTRACT: Wood-boring bivalves (families Xylophagaidae and Teredinidae) are unique bivalves that are peculiarly adapted to feed on terrigenous woody materials, even though they inhabit the deep-sea floor far from land. Previous studies of their metabolic processes suggested the importance of symbiotic microbes that secrete cellulase to access woody carbon (carbohydrates), and supply organic nitrogen via nitrogen fixation. Since nitrogen is generally depleted in woody materials, dissolved dinitrogen in seawater has been proposed as a plausible nitrogen source for wood-boring bivalves. We evaluated the food ecology of wood-boring bivalves (genus: Xyloredo) obtained from the deep seafloor off the Ryukyu Islands by considering their bulk carbon, nitrogen, and sulfur isotope composition and potential dietary sources (i.e. logs, particulate organic matter, and surface sediments). We also investigated the trophic interactions between wood-boring bivalves and logs based on the amino acid nitrogen isotopic composition. The bulk isotope data revealed that in wood-boring bivalves these elements are derived mainly from the logs in which they live. These results were consistent with the trophic hierarchy calculated from the nitrogen isotopic composition of amino acids. Ecologically, wood-boring bivalves are one step higher than logs in terms of their trophic position. Based on these data, we propose that terrigenous woody materials are the major dietary sources for wood-boring bivalves and the same standard trophic interaction exists between diet and consumer species as in the grazing food web. The symbionts may aid the digestion of woody materials, but they do not supply nitrogen via nitrogen fixation in this case. KEY WORDS: Wood-boring bivalves · Xyloredo teramachii · Compound-specific nitrogen isotope analysis · Amino acids · Nitrogen nutrition · Stable isotope composition Full text in pdf format PreviousNextCite this article as: Yamanaka T, Shimamura S, Chikaraishi Y, Haga T, Fujiwara Y (2015) Re-evaluation of nutrient sources for deep-sea wood-boring bivalves using the isotopic composition of bulk C, N, S, and amino acid nitrogen. Mar Ecol Prog Ser 540:157-165. https://doi.org/10.3354/meps11510 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 540. Online publication date: November 26, 2015 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2015 Inter-Research.