Amino Acid 15N/14N Analysis at Natural Abundances: A New Tool for Soil Organic Matter Studies in Agricultural Systems

Abstract

The effects of landuse, fertilizer history and soil type on the quantity and isotopic quality of hydrolysable soil amino acids were examined in 3 grassland and 2 arable soils. Results showed, (i) that overall concentrations of individual amino acids were highest in the grassland soils, (ii) that ‰δ¹⁵N values of the individual amino acids differed considerably between the five soils, and (iii) that the combination of amino acid ‰δ¹⁵N values and concentrations could be used to distinguish between landuse, crop type and fertilizer history. This preliminary study indicates that the pathways of transformation of soil amino acid N are influenced by long term N inputs and that associated biological processes are reflected in differences in concentrations and ‰δ¹⁵N values of individual soil amino acids.     

Long term changes in the distribution and delta(15)N values of individual soil amino acids in the absence of plant and fertiliser inputs

The long-term 'biodegradation' on soil amino acids was examined in the control plots of '42 parcelles' experiment, established in 1928 at INRA, Versailles (France). None of the plots is cultivated, but is kept free of weeds, and mixed to a depth of 25 cm twice yearly. Topsoil (0-10 cm depth) samples collected in 1929, 1963 and 1997 were subjected to acid hydrolysis (6 N HCl) for comparison. The distribution and delta(15)N natural abundance of 20 individual amino acids in the soils were determined, using ion chromatography (IC) and gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). The total N and amino acid-N (AA-N), respectively, decreased by 54 % and 73 % in the period from 1929 to 1997. The average N loss was comparable for 1929-1963 (period 1) and 1963-1997 (period 2), but AA-N loss was three times faster in the former period. This significant reduction in total AA-N content was mirrored in the individual amino acids, which decreased by 74 % +/- 1 % (ranging 58-89 %) between 1929 and 1997. The bulk delta(15)N values generally increased from 1929 to 1997, mainly associated with comparable or even higher increase of delta(15)N of the non-AA-N in the soil. The residence time (t(1/2), time in which half of N was lost from a specific soil pool) was ca. 65 +/- 5 years for the bulk soil, and comparable for periods 1 and 2. However, between periods 1 and 2 it decreased from 128 to 41 years in the non-AA pool, but increased from 59 to 92 years in the AA-N pool. Proline and amino acids that appear early in soil microbial metabolic pathways (e.g. glutamic acid, alanine, aspartic acid and valine) had relatively high delta(15)N values. Phenylalanine, threonine, glycine and leucine had relatively depleted delta(15)N values. The average delta(15)N value of the individual amino acids (IAAs) increased by 1delta unit from 1929 to 1997, associated with a similar rise from 1929 to 1963, and no change thereafter till 1997. However, the delta(15)N values of phenylalanine decreased by more than 7delta(15)N units between 1929 and 1997. The delta(15)N shift of IAAs from 1929 to 1963 and from 1929 to 1997 was not influenced by the relative amount of N remaining compared with the 1929 soil concentrations. The only exception was phenylalanine which showed decreasing delta(15)N associated with its decreasing concentration in the soil. We conclude therefore that in the absence of plant and fertiliser inputs, no change in the delta(15)N value of individual soil amino acids occurs, hence the original delta(15)N values are preserved and diagnostic information on past soil N (cycling) is retained. The exception was phenylalanine, its delta(15)N decreased with decreasing concentration from 1929 to 1997, hence it acted as a 'potential' marker for the land use changes (i.e. arable cropping to a fallow). The long term biological processing and reworking of residual amino acids resulted in a (partial) stabilisation in the soil, evidenced by reduced N loss and increased residence time of amino acid N during the period 1963-1997.     

Long term changes in the distribution and δ15N values of individual soil amino acids in the absence of plant and fertiliser inputs

The long-term ‘biodegradation’ on soil amino acids was examined in the control plots of ‘42 parcelles’ experiment, established in 1928 at INRA, Versailles (France). None of the plots is cultivated, but is kept free of weeds, and mixed to a depth of 25 cm twice yearly. Topsoil (0–10 cm depth) samples collected in 1929, 1963 and 1997 were subjected to acid hydrolysis (6 N HCl) for comparison. The distribution and δ¹⁵N natural abundance of 20 individual amino acids in the soils were determined, using ion chromatography (IC) and gas chromatography–combustion–isotope ratio mass spectrometry (GC–C–IRMS). The total N and amino acid-N (AA-N), respectively, decreased by 54 % and 73 % in the period from 1929 to 1997. The average N loss was comparable for 1929–1963 (period 1) and 1963–1997 (period 2), but AA-N loss was three times faster in the former period. This significant reduction in total AA-N content was mirrored in the individual amino acids, which decreased by 74 %?±?1 % (ranging 58–89 %) between 1929 and 1997. The bulk δ¹⁵N values generally increased from 1929 to 1997, mainly associated with comparable or even higher increase of δ¹⁵N of the non-AA-N in the soil. The residence time (t _1/2, time in which half of N was lost from a specific soil pool) was ca. 65?±?5 years for the bulk soil, and comparable for periods 1 and 2. However, between periods 1 and 2 it decreased from 128 to 41 years in the non-AA pool, but increased from 59 to 92 years in the AA-N pool. Proline and amino acids that appear early in soil microbial metabolic pathways (e.g. glutamic acid, alanine, aspartic acid and valine) had relatively high δ¹⁵N values. Phenylalanine, threonine, glycine and leucine had relatively depleted δ¹⁵N values. The average δ¹⁵N value of the individual amino acids (IAAs) increased by 1δ unit from 1929 to 1997, associated with a similar rise from 1929 to 1963, and no change thereafter till 1997. However, the δ¹⁵N values of phenylalanine decreased by more than 7δ¹⁵N units between 1929 and 1997. The δ¹⁵N shift of IAAs from 1929 to 1963 and from 1929 to 1997 was not influenced by the relative amount of N remaining compared with the 1929 soil concentrations. The only exception was phenylalanine which showed decreasing δ¹⁵N associated with its decreasing concentration in the soil. We conclude therefore that in the absence of plant and fertiliser inputs, no change in the δ¹⁵N value of individual soil amino acids occurs, hence the original δ¹⁵N values are preserved and diagnostic information on past soil N (cycling) is retained. The exception was phenylalanine, its δ¹⁵N decreased with decreasing concentration from 1929 to 1997, hence it acted as a ‘potential’ marker for the land use changes (i.e. arable cropping to a fallow). The long term biological processing and reworking of residual amino acids resulted in a (partial) stabilisation in the soil, evidenced by reduced N loss and increased residence time of amino acid N during the period 1963–1997.     

Interpreting early land management through compound specific stable isotope analyses of archaeological soils

Compound specific stable isotope analyses of managed soils using isotope ratio mass spectrometry have been undertaken as a means of determining early land use practices. delta (15)N amino acid signals demonstrate differences between manured grassland, unmanured grassland and continuous cereal cultivation under long-term experimental land use control conditions, with delta (15)N in hydrophobic amino acids providing the most distinctive signals. Analysis of early modern/medieval and of Bronze age anthropogenic soils from Orkney demonstrates that such signals are retained in archaeological contexts. delta (13)C analyses of n- alkanoic acid components of the fossil, Bronze Age, anthropogenic soils suggest a major terrestrial input to these soils, with uniform composition of formation materials. Surficial soils demonstrate the assimilation of isotopically lighter carbon, providing a means of assessing the mobility of the n- alkanoic acids within soils and sediments. Copyright 1999 John Wiley & Sons, Ltd.     

Oxygen and nitrogen isotopic composition of nitrate in commercial fertilizers,nitric acid,and reagent salts

Nitrate is a key component of synthetic fertilizers that can be beneficial to crop production in agro-ecosystems, but can also cause damage to natural ecosystems if it is exported in large amounts. Stable isotopes, both oxygen and nitrogen, have been used to trace the sources and fate of nitrate in various ecosystems. However, the oxygen isotope composition of synthetic and organic nitrates is poorly constrained. Here, we present a study on the N and O isotope composition of nitrate-based fertilizers. The δ¹⁵N values of synthetic and natural nitrates were 0?±?2?‰ similar to the air N₂ from which they are derived. The δ¹⁸O values of synthetic nitrates were 23?±?3?‰, similar to air O₂, and natural nitrate fertilizer δ¹⁸O values (55?±?5?‰) were similar to those observed in atmospheric nitrate. The Δ¹⁷O values of synthetic fertilizer nitrate were approximately zero following a mass-dependent isotope relationship, while natural nitrate fertilizers had Δ¹⁷O values of 18?±?2?‰ similar to nitrate produced photochemically in the atmosphere. These narrow ranges of values can be used to assess the amount of nitrate arising from fertilizers in mixed systems where more than one nitrate source exists (soil, rivers, and lakes) using simple isotope mixing models.     

Seasonal variation in natural abundance of δ13C and 15N in Salicornia brachiata Roxb. populations from a coastal area of India

High and fluctuating salinity is characteristic for coastal salt marshes, which strongly affect the physiology of halophytes consequently resulting in changes in stable isotope distribution. The natural abundance of stable isotopes (δ¹³C and δ¹⁵N) of the halophyte plant Salicornia brachiata and physico-chemical characteristics of soils were analysed in order to investigate the relationship of stable isotope distribution in different populations in a growing period in the coastal area of Gujarat, India. Aboveground and belowground biomass of S. brachiata was collected from six different populations at five times (September 2014, November 2014, January 2015, March 2015 and May 2015). The δ¹³C values in aboveground (?30.8 to ?23.6?‰, average: ?26.6?±?0.4?‰) and belowground biomass (?30.0 to ?23.1?‰, average: ?26.3?±?0.4?‰) were similar. The δ¹³C values were positively correlated with soil salinity and Na concentration, and negatively correlated with soil mineral nitrogen. The δ¹⁵N values of aboveground (6.7–16.1?‰, average: 9.6?±?0.4?‰) were comparatively higher than belowground biomass (5.4–13.2?‰, average: 7.8?±?0.3?‰). The δ¹⁵N values were negatively correlated with soil available P. We conclude that the variation in δ¹³C values of S. brachiata was possibly caused by soil salinity (associated Na content) and N limitation which demonstrates the potential of δ¹³C as an indicator of stress in plants.     

Spatial variations in larch needle and soil δ15N at a forest–grassland boundary in northern Mongolia

The spatial patterns of plant and soil δ¹⁵N and associated processes in the N cycle were investigated at a forest–grassland boundary in northern Mongolia. Needles of Larix sibirica Ledeb. and soils collected from two study areas were analysed to calculate the differences in δ¹⁵N between needle and soil (Δδ¹⁵N). Δδ¹⁵N showed a clear variation, ranging from ?8?‰ in the forest to ?2?‰ in the grassland boundary, and corresponded to the accumulation of organic layer. In the forest, the separation of available N produced in the soil with ¹⁵N-depleted N uptake by larch and ¹⁵N-enriched N immobilization by microorganisms was proposed to cause large Δδ¹⁵N, whereas in the grassland boundary, small Δδ¹⁵N was explained by the transport of the most available N into larch. The divergence of available N between larch and microorganisms in the soil, and the accumulation of diverged N in the organic layer control the variation in Δδ¹⁵N.     

Human and climate impact on ¹⁵N natural abundance of plants and soils in high-mountain ecosystems: a short review and two examples from the Eastern Pamirs and Mt. Kilimanjaro

Population pressure increasingly endangers high-mountain ecosystems such as the pastures in the Eastern Pamirs and the mountain forests on Mt. Kilimanjaro. At the same time, these ecosystems constitute the economic basis for millions of people living there. In our study, we, therefore, aimed at characterising the land-use effects on soil degradation and N-cycling by determining the natural abundance of (15)N. A short review displays that δ(15)N of plant-soil systems may often serve as an integrated indicator of N-cycles with more positive δ(15)N values pointing towards N-losses. Results for the high-mountain pastures in the Eastern Pamirs show that intensively grazed pastures are significantly enriched in (15)N compared to the less-exploited pastures by 3.5 ‰, on average. This can be attributed to soil organic matter degradation, volatile nitrogen losses, nitrogen leaching and a general opening of the N-cycle. Similarly, the intensively degraded savanna soils, the cultivated soils and the soils under disturbed forests on the foothill of Mt. Kilimanjaro reveal very positive δ(15)N values around 6.5 ‰. In contrast, the undisturbed forest soils in the montane zone are more depleted in (15)N, indicating that here the N-cycle is relatively closed. However, significantly higher δ(15)N values characterise the upper montane forest zone at the transition to the subalpine zone. We suggest that this reflects N-losses by the recently monitored and climate change and antropogenically induced increasing fire frequency pushing the upper montane rainforest boundary rapidly downhill. Overall, we conclude that the analysis of the (15)N natural abundance in high-mountain ecosystems is a purposeful tool for detecting land-use- or climate change-induced soil degradation and N-cycle opening.     

Differential response of two Pinus spp. to avian nitrogen input as revealed by nitrogen isotope analysis for tree rings

Temporal variations in N concentration and δ(15)N value of annual tree rings (1 year of time resolution) of two Japanese Black Pine (Pinus thunbergii) and three Japanese Red Pine (Pinus densiflora) trees under current breeding activity of the Great Cormorant (Pharacrocorax carbo) and the Black-tailed Gull (Larus crassirostris), respectively, in central and northeastern Japan were studied. Both species from control sites where no avian input occurs show negative values (δ(15)N = around -4 ‰ to -2 ‰) which are common among higher plants growing under high rainfall regimes. The δ(15)N values of P. densiflora show uniformly positive values several years before and after the breeding event, indicating N translocation that moved the absorbed N of a given growth year to tree rings of the previous year while a clear historical value of soil N dynamics was kept intact in the annual rings of P. thunbergii. Long-term N trends inferred from tree rings must take into account tree species with limited translocation rates that can retain actual N annual acquisition.     

不同施肥处理对潮土胡敏酸结构特性的影响

研究了饼肥,绿肥,秸秆肥三种处理在等氮等磷条件下对胡敏酸结构特性的影响,采用元素分析,显微红外,固态13C-NMR三种分析方法进行结构解析,结果表明：不同施肥处理的胡敏酸在化学组成和结构上类似,但还有很多明显的差别。(1)饼肥处理的胡敏酸H和N含量最高,绿肥处理的胡敏酸含O量最大,秸秆肥处理的胡敏酸含C量最高 (2)红外分析显示,三种处理的胡敏酸均含有蛋白质及碳水化合物,饼肥处理的胡敏酸含氨基化合物最多,绿肥处理的胡敏酸含羟基,脂肪烃含量最大,秸秆肥处理的胡敏酸含醇,酚含量最高;(3)固态 13C-NMR数据表明饼肥处理的胡敏酸含有最多的羧基碳,绿肥处理的胡敏酸含有烷基碳,羰基碳最多。秸秆肥处理的胡敏酸烷氧基碳,芳香碳含量最大。     

Diskriminierung der Stickstoff-Isotope im Leber-Metabolismus nach Verzehr von Unterschiedlichen Nahrungsproteinen

Nitrogen isotope ratio disproportionation (N-IRD) is the result of isotope effects of different magnitude in branched pathways of nitrogen metabolism of the liver. N-IRD is measured as the difference of natural ¹⁵N abundance of urea and plasma protein and is noted in ‰ (per mill) as the difference of their δ values.

It was derived that N-IRD would become maximum if the amino acid pattern consumed by the respective pathways urea synthesis and protein synthesis of the liver is identical. This is expected if the metabolic available amino acid pattern exactly meets requirement.

When feeding rats methionine(met)-supplemented casein N-IRD increased from 6‰ at zero protein intake to 8‰ at the requirement intake of 5 g/kg BM?)/day. With further increasing protein intake N-IRD decreased to values below 6‰. It was concluded that increasing N-IRD with increasing intake of met-supplemented casein is caused by complementation of the endogenous amino acid pattern by exogenous dietary amino acids with a complementation optimum at protein intake of 5 g/kg BM/day where N-IRD was maximum.

When feeding rats soy protein N-IRD (= 6.2‰) did not increase, but declined slightly above requirement intake of 10 g/kg BM/day to N-IRD = 5.7‰ at 30 g/kg BM/day. This indicates that the amino acid pattern of the metabolic pool was not complemented by intake of soy protein.

These findings strongly suggest that the (endogenous) amino acid pattern of the metabolic pool, as present at zero protein intake, is characterized by an imbalance (“endogenous inbalance”) which may be complemented dependent on the intake of appropriate protein.     

Human and climate impact on 15N natural abundance of plants and soils in high-mountain ecosystems: a short review and two examples from the Eastern Pamirs and Mt. Kilimanjaro

Population pressure increasingly endangers high-mountain ecosystems such as the pastures in the Eastern Pamirs and the mountain forests on Mt. Kilimanjaro. At the same time, these ecosystems constitute the economic basis for millions of people living there. In our study, we, therefore, aimed at characterising the land-use effects on soil degradation and N-cycling by determining the natural abundance of ¹⁵N. A short review displays that δ¹⁵N of plant–soil systems may often serve as an integrated indicator of N-cycles with more positive δ¹⁵N values pointing towards N-losses. Results for the high-mountain pastures in the Eastern Pamirs show that intensively grazed pastures are significantly enriched in ¹⁵N compared to the less-exploited pastures by 3.5 ‰, on average. This can be attributed to soil organic matter degradation, volatile nitrogen losses, nitrogen leaching and a general opening of the N-cycle. Similarly, the intensively degraded savanna soils, the cultivated soils and the soils under disturbed forests on the foothill of Mt. Kilimanjaro reveal very positive δ¹⁵N values around 6.5 ‰. In contrast, the undisturbed forest soils in the montane zone are more depleted in ¹⁵N, indicating that here the N-cycle is relatively closed. However, significantly higher δ¹⁵N values characterise the upper montane forest zone at the transition to the subalpine zone. We suggest that this reflects N-losses by the recently monitored and climate change and antropogenically induced increasing fire frequency pushing the upper montane rainforest boundary rapidly downhill. Overall, we conclude that the analysis of the ¹⁵N natural abundance in high-mountain ecosystems is a purposeful tool for detecting land-use- or climate change-induced soil degradation and N-cycle opening.     

The relative isotopic abundance (δ13C, δ15N) during composting of agricultural wastes in relation to compost quality and feedstock

Variations in the relative isotopic abundance of C and N (δ¹³C and δ¹⁵N) were measured during the composting of different agricultural wastes using bench-scale bioreactors. Different mixtures of agricultural wastes (horse bedding manure?+?legume residues; dairy manure?+?jatropha mill cake; dairy manure?+?sugarcane residues; dairy manure alone) were used for aerobic–thermophilic composting. No significant differences were found between the δ¹³C values of the feedstock and the final compost, except for dairy manure?+?sugarcane residues (from initial ratio of ?13.6?±?0.2?‰ to final ratio of ?14.4?±?0.2?‰). δ¹⁵N values increased significantly in composts of horse bedding manure?+?legumes residues (from initial ratio of +5.9?±?0.1?‰ to final ratio of +8.2?±?0.5?‰) and dairy manure?+?jatropha mill cake (from initial ratio of +9.5?±?0.2?‰ to final ratio of +12.8?±?0.7?‰) and was related to the total N loss (mass balance). δ¹³C can be used to differentiate composts from different feedstock (e.g. C₃ or C₄ sources). The quantitative relationship between N loss and δ¹⁵N variation should be determined.     

A dynamic study of earthworm feeding ecology -using stable isotopes

Changes in the specific diet of earthworms with time in relation to landuse changes and two different climates were studied by analysing (13)C and (15)N natural abundance in soils and animals. Soil samples from three depths (0-10, 10-20 and 20-30 cm) and earthworms were collected from two sites: Santiago (Northwest Spain) and North Wyke (Southwest England) both consisting of replicated long-term grasslands and recently converted to maize plots. Earthworms were hand-sorted in the field at the peak of the maize growth and after harvesting at both sites. In the Spanish plots, nine and eight earthworm species, all belonging to the three ecological categories (epigeic, anecic and endogeic), were found under maize and permanent pasture, whereas at the English site five and seven different species were, respectively, identified. At both sites (13)C isotopic values of the earthworm tissues reflected changes in diet from C(3) to C(4) with epigeic and epi/anecic worms in the maize plots showing one delta unit difference in relation to the ones found in the grassland plots. Anecic worms seemed to be less responsive to landuse changes. The higher (13)C values of the Spanish soils were also reflected in the earthworm tissues when compared with the English samples. (15)N values showed no clear relationship with the cropping treatments but were clearly related to the ecological grouping, with endogeic worms reaching the highest values whereas for the epigeic and epi/anecic species the lowest values were obtained. This finding was also previously recorded by other authors1 and suggests that, in the future, stable isotope techniques could also be a useful tool in taxonomic studies. Copyright 1999 John Wiley & Sons, Ltd.     

Short-term changes in delta(13)C and delta(15)N signatures of water discharged from grazed grasslands

The composition of dissolved organic matter (DOM) in a soil is the product of a variety of soil processes. Changes in the composition of DOM in water discharged from soil should, therefore, give an important insight into modifications in these soil processes. We hypothesise that these processes in soils, under different grassland management regimes, would be affected to different extents by the short-term disturbance of a storm event and that evidence of this could be detected in delta(13)C and delta(15)N signatures in drainage and surface runoff waters. During a storm event we collected discharge waters from 1 ha grassland lysimeters, with or without artificial drainage, which received contrasting fertiliser inputs, and delta(13)C and delta(15)N signatures were determined. Changes in (13)C enrichment during the storm event were clearly identifiable, as were differences between plots for (13)C and (15)N, illustrating that this technique has potential to be a useful tool for identifying and investigating short- and long-term changes in soil organic matter dynamics. Copyright 1999 John Wiley & Sons, Ltd.     

Characteristics of suspended matter in the River Sava watershed,Slovenia

A combination of C/N ratios, δ¹³C and δ¹⁵N values in suspended matter was used to examine the seasonal (late summer 2004 and spring 2005) relationship with hydrological characteristics of the River Sava watershed in Slovenia. The values of C/N ratios range from 1.2 to 19.1, δ¹³C values range from?29.2 to?23.0 ‰ and δ¹⁵N values from 0.5 to 16.7 ‰ and indicate that the samples are a mixture of two end members: modern soils and plant litter. A simple mixing model was used to indicate that soil organic carbon prevails over plant litter and contributes more than 50% of suspended material. The calculated annual particulate organic carbon flux is estimated as 5.2×10¹⁰ g C/year, the annual particulate nitrogen flux 8.5×10⁹ g N/year and the total suspended solid flux is estimated to be 1.3×10¹² g/year. Anthropogenic impact was detected only in a tributary stream of the River Sava, which is located in an agriculture–industrial area and is reflected in higher δ¹⁵N values in suspended matter and high nitrate concentrations in the late summer season.     

Isotope (δ13C, δ15N, δ2H) diet–tissue discrimination in African grey parrot Psittacus erithacus: implications for forensic studies

Diet–tissue isotopic relationships established under controlled conditions are informative for determining the dietary sources and geographic provenance of organisms. We analysed δ¹³C, δ¹⁵N, and non-exchangeable δ²H values of captive African grey parrot Psittacus erithacus feathers grown on a fixed mixed-diet and borehole water. Diet–feather Δ¹³C and Δ¹⁵N discrimination values were +3.8?±?0.3?‰ and +6.3?±?0.7?‰ respectively; significantly greater than expected. Non-exchangeable δ²H feather values (?62.4?±?6.4?‰) were more negative than water (?26.1?±?2.5?‰) offered during feather growth. There was no positive relationship between the δ¹³C and δ¹⁵N values of the samples along each feather with the associated samples of food offered, or the feather non-exchangeable hydrogen isotope values with δ²H values of water, emphasising the complex processes involved in carbohydrate, protein, and income water routing to feather growth. Understanding the isotopic relationship between diet and feathers may provide greater clarity in the use of stable isotopes in feathers as a tool in determining origins of captive and wild-caught African grey parrots, a species that is widespread in aviculture and faces significant threats to wild populations. We suggest that these isotopic results, determined even in controlled laboratory conditions, be used with caution.     

Differential response of two Pinus spp. to avian nitrogen input as revealed by nitrogen isotope analysis for tree rings

Temporal variations in N concentration and δ¹⁵N value of annual tree rings (1 year of time resolution) of two Japanese Black Pine (Pinus thunbergii) and three Japanese Red Pine (Pinus densiflora) trees under current breeding activity of the Great Cormorant (Pharacrocorax carbo) and the Black-tailed Gull (Larus crassirostris), respectively, in central and northeastern Japan were studied. Both species from control sites where no avian input occurs show negative values (δ¹⁵N = around?4 ‰ to?2 ‰) which are common among higher plants growing under high rainfall regimes. The δ¹⁵N values of P. densiflora show uniformly positive values several years before and after the breeding event, indicating N translocation that moved the absorbed N of a given growth year to tree rings of the previous year while a clear historical value of soil N dynamics was kept intact in the annual rings of P. thunbergii. Long-term N trends inferred from tree rings must take into account tree species with limited translocation rates that can retain actual N annual acquisition.     

Field-deployable measurements of free-living individuals to determine energy balance: fuel substrate usage through δ13C in breath CO2 and diet through hair δ13C and δ15N values

Carbon isotopes of breath CO₂ vary depending on diet and fuel substrate used. This study examined if exercise-induced δ¹³C-CO₂ changes in substrate utilization were distinguishable from baseline δ¹³C-CO₂ variations in a population with uncontrolled diet, and compared hair isotope values and food logs to develop an isotope model of diet. Study participants included nine women with diverse Body Mass Index (BMI), age, ancestry, exercise history, and diet. Breath samples were collected prior to and up to 12?h after a 5- or 10?K walk/run. Indirect calorimetry was measured with a smartphone-enabled mobile colorimetric device, and a field-deployable isotope analyzer measured breath δ¹³C-CO₂ values. Diet was assessed by food logs and δ¹³C, δ¹⁵N of hair samples. Post-exercise δ¹³C-CO₂ values increased by 0.54?±?1.09‰ (1 sd, n?=?9), implying enhanced carbohydrate burning, while early morning δ¹³C-CO₂ values were lower than daily averages (p?=?0.0043), indicating lipid burning during overnight fasting. Although diurnal δ¹³C-CO₂ variation (1.90?±?0.77‰) and participant baseline range (3.06‰) exceeded exercise-induced variation, temporal patterns distinguished exercise from dietary isotope effects. Hair δ¹³C and δ¹⁵N values were consistent with a new dietary isotope model. Notwithstanding the small number of participants, this study introduces a novel combination of techniques to directly monitor energy balance in free-living individuals.     

Latitudinal distribution of nitrogen isotopic composition in suspended particulate organic matter in tropical/subtropical seas

Natural nitrogen isotopic composition (δ(15)N) of suspended particulate organic matter (POM) and nitrogen fixation rates via (15)N(2) assay were measured in surface waters along 120° E from 30° N to 30° S in the Asian marginal seas (the East/South China Seas and the Sulu/Celebes/Java Seas) and the northeastern Indian Ocean in November-December 2005 and March 2006. The POM δ(15)N values ranged from-1.8 to 12.2‰ with an average of 3.6‰ and showed a decreasing trend towards the equator in both hemispheres. In parallel, the measured N(2) fixation rates showed an increase from the subtropical to the tropical seas. This implies that a higher contribution of (15)N-depleted POM was derived from enhanced N(2) fixation. Water temperature and the stability of water column were partly responsible for the observed variations in nitrogen fixation. The large-scale spatial variations in suspended POM δ(15)N and N(2) fixation rates suggest that the suspended POM δ(15)N may be a potential indicator of nitrogen fixation in surface waters in tropical/subtropical seas.