Variations in microbial isotopic fractionation during soil organic matter decomposition

The soil microbial biomass (SMB) is known to participate in key soil processes such as the decomposition of soil organic matter (SOM). However, its contribution to the isotopic composition of the SOM is not clear yet. Shifts in the ¹³C and ¹⁵N natural abundances of the SMB and SOM fractions (mineralised, water soluble and non-extractable) were investigated by incubating an unamended arable soil for 6 months. Microbial communities were also studied using Fatty Acid Methyl Ester specific isotope analysis. The SMB was significantly ¹³C and ¹⁵N-enriched relative to other fractions throughout the incubation. However, significant isotopic variations with time were also observed due to the rapid consumption of relatively ¹³C-enriched water soluble compounds. The increase in the difference in SMB and water soluble ¹⁵N compositions as the water soluble C/N ratio decreased, indicated a shift from N assimilation to N dissimilation during the incubation. These changes also induced modifications of the microbial community structure. Once the system reached a steady-state (after 1 month), the isotopic trends appeared to corroborate those obtained in long term experiments in the field in that there was a constant microbial isotopic fractionation leading to a ¹³C and ¹⁵N enrichment of the SOM over the long-term. This work also suggests that caution must be exercised when interpreting short term incubation studies since perturbations associated with experimental set-up can have an important effect on C and N dynamics, microbial fractionation of ¹³C and ¹⁵N and microbial community structure.     

Effect of C3–C4 Vegetation Change on δ13C and δ15N Values of Soil Organic Matter Fractions Separated by Thermal Stability

Carbon isotopic composition of soils subjected to C₃–C₄ vegetation change can be used to estimate C turnover in bulk soil and in soil organic matter (SOM) pools with fast and intermediate turnover rates. We hypothesized that the biological availability of SOM pools is inversely proportional to their thermal stability, so that thermogravimetry can be used to separate SOM pools with contrasting turnover rates. Soil samples from a field plot cultivated for 10.5 years with the perennial C₄ plant Miscanthus×gigantheus were analyzed by thermogravimetry coupled with differential scanning calorimetry (DSC). Three SOM fractions were distinguished according to the differential weight losses and exothermic or endothermic reactions measured by DSC. The δ¹³C and δ¹⁵N values of these three fractions obtained by gradual soil heating were measured by IRMS. The weight losses up to 190 °C mainly reflected water evaporation because no significant C and N losses were detected and δ¹³C and δ¹⁵N values of the residual SOM remained unchanged. The δ¹³C values (−16.4‰) of SOM fraction decomposed between 190 and 390 °C (containing 79% of total soil C) were slightly closer to that of the Miscanthus plant tissues (δ¹³C = −11.8‰) compared to the δ¹³C values (−16.8‰) of SOM fraction decomposed above 390 °C containing the residual 21% of SOM. Thus, the C turnover in the thermally labile fraction was faster than that in thermally stable fractions, but the differences were not very strong. Therefore, in this first study combining TG-DSC with isotopic analysis, we conclude that the thermal stability of SOM was not very strongly related to biological availability of SOM fractions. In contrast to δ¹³C, the δ¹⁵N values strongly differed between SOM fractions, suggesting that N turnover in the soil was different from C turnover. More detailed fractionation of SOM by thermal analysis with subsequent isotopic analysis may improve the resolution for δ¹³C.     

Transport of Carbon and Nitrogen Between Litter and Soil Organic Matter in a Northern Hardwood Forest

We used sugar maple litter double-labeled with ¹³C and ¹⁵N to quantify fluxes of carbon (C) and nitrogen (N) between litter and soil in a northern hardwood forest and the retention of litter C and N in soil. Two cohorts of litter were compared, one in which the label was preferentially incorporated into non-structural tissue and the other structural tissue. Loss of ¹³C from this litter generally followed dry mass and total C loss whereas loss of ¹⁵N (20–30% in 1 year) was accompanied by large increases of total N content of this decaying litter (26–32%). Enrichment of ¹³C and ¹⁵N was detected in soil down to 10–15 cm depth. After 6 months of decay (November–May) 36–43% of the ¹³C released from the litter was recovered in the soil, with no differences between the structural and non-structural labeled litter. By October the percentage recovery of litter ¹³C in soil was much lower (16%). The C released from litter and remaining in soil organic matter (SOM) after 1 year represented over 30 g C m⁻² y⁻¹ of SOM accumulation. Recovery of litter ¹⁵N in soil was much higher than for C (over 90%) and in May ¹⁵N was mostly in organic horizons whereas by October it was mostly in 0–10 cm mineral soil. A small proportion of this N was recovered as inorganic N (2–6%). Recovery of ¹⁵N in microbial biomass was higher in May (13–15%) than in October (about 5%). The C:N ratio of the SOM and microbial biomass derived from the labeled litter was much higher for the structural than the non-structural litter and for the forest floor than mineral SOM, illustrating the interactive role of substrates and microbial activity in regulating the C:N stoichiometry of forest SOM formation. These results for a forest ecosystem long exposed to chronically high atmospheric N deposition (ca. 10 kg N ha⁻¹ y⁻¹) suggest possible mechanisms of N retention in soil: increased organic N leaching from fresh litter and reduced fungal transport of N from soil to decaying litter may promote N stabilization in mineral SOM even at a relatively low C:N ratio.     

Variability and directionality of temporal changes in δ<Superscript>13</Superscript>C and δ<Superscript>15</Superscript>N of aquatic invertebrate primary consumers

Seasonal oscillations in the carbon (δ¹³C) and nitrogen (δ¹⁵N) isotope signatures of aquatic algae can cause seasonal enrichment–depletion cycles in the isotopic composition of planktonic invertebrates (e.g., copepods). Yet, there is growing evidence that seasonal enrichment–depletion cycles also occur in the isotope signatures of larger invertebrate consumers, taxa used to define reference points in isotope-based trophic models (e.g., trophic baselines). To evaluate the general assumption of temporal stability in non-zooplankton aquatic invertebrates, δ¹³C and δ¹⁵N time series data from the literature were analyzed for seasonality and the influence of biotic (feeding group) and abiotic (trophic state, climate regime) factors on isotope temporal patterns. The amplitude of δ¹³C and δ¹⁵N enrichment–depletion cycles was negatively related to body size, although all size-classes of invertebrates displayed a winter-to-summer enrichment in δ¹³C and depletion in δ¹⁵N. Among feeding groups, periphytic grazers were more variable and displayed larger temporal changes in δ¹³C than detritivores. For nitrogen, temporal variability and magnitude of directional change of δ¹⁵N was most strongly related to ecosystem trophic state (eutrophic > mesotrophic, oligotrophic). This study provides evidence of seasonality in the isotopic composition of aquatic invertebrates across very broad geographical and ecological gradients as well as identifying factors that are likely to modulate the strength and variability of seasonality. These results emphasize the need for researchers to recognize the likelihood of temporal changes in non-zooplankton aquatic invertebrate consumers at time scales relevant to seasonal studies and, if present, to account for temporal dynamics in isotope trophic models.     

Isotope fractionation and 13C enrichment in soil profiles during the decomposition of soil organic matter

The mechanisms behind the ¹³C enrichment of organic matter with increasing soil depth in forests are unclear. To determine if ¹³C discrimination during respiration could contribute to this pattern, we compared δ¹³C signatures of respired CO₂ from sieved mineral soil, litter layer and litterfall with measurements of δ¹³C and δ¹⁵N of mineral soil, litter layer, litterfall, roots and fungal mycelia sampled from a 68-year-old Norway spruce forest stand planted on previously cultivated land. Because the land was subjected to ploughing before establishment of the forest stand, shifts in δ¹³C in the top 20 cm reflect processes that have been active since the beginning of the reforestation process. As ¹³C-depleted organic matter accumulated in the upper soil, a 1.0‰ δ¹³C gradient from −28.5‰ in the litter layer to −27.6‰ at a depth of 2–6 cm was formed. This can be explained by the 1‰ drop in δ¹³C of atmospheric CO₂ since the beginning of reforestation together with the mixing of new C (forest) and old C (farmland). However, the isotopic change of the atmospheric CO₂ explains only a portion of the additional 1.0‰ increase in δ¹³C below a depth of 20 cm. The δ¹³C of the respired CO₂ was similar to that of the organic matter in the upper soil layers but became increasingly ¹³C enriched with depth, up to 2.5‰ relative to the organic matter. We hypothesise that this ¹³C enrichment of the CO₂ as well as the residual increase in δ¹³C of the organic matter below a soil depth of 20 cm results from the increased contribution of ¹³C-enriched microbially derived C with depth. Our results suggest that ¹³C discrimination during microbial respiration does not contribute to the ¹³C enrichment of organic matter in soils. We therefore recommend that these results should be taken into consideration when natural variations in δ¹³C of respired CO₂ are used to separate different components of soil respiration or ecosystem respiration.     

Soil microbial diversity affects soil organic matter decomposition in a silty grassland soil

Soil microorganisms play a pivotal role in soil organic matter (SOM) turn-over and their diversity is discussed as a key to the function of soil ecosystems. However, the extent to which SOM dynamics may be linked to changes in soil microbial diversity remains largely unknown. We characterized SOM degradation along a microbial diversity gradient in a two month incubation experiment under controlled laboratory conditions. A microbial diversity gradient was created by diluting soil suspension of a silty grassland soil. Microcosms containing the same sterilized soil were re-inoculated with one of the created microbial diversities, and were amended with ¹³C labeled wheat in order to assess whether SOM decomposition is linked to soil microbial diversity or not. Structural composition of wheat was assessed by solid-state ¹³C nuclear magnetic resonance, sugar and lignin content was quantified and labeled wheat contribution was determined by ¹³C compound specific analyses. Results showed decreased wheat O-alkyl-C with increasing microbial diversity. Total non-cellulosic sugar-C derived from wheat was not significantly influenced by microbial diversity. Carbon from wheat sugars (arabinose-C and xylose-C), however, was highest when microbial diversity was low, indicating reduced wheat sugar decomposition at low microbial diversity. Xylose-C was significantly correlated with the Shannon diversity index of the bacterial community. Soil lignin-C decreased irrespective of microbial diversity. At low microbial diversity the oxidation state of vanillyl–lignin units was significantly reduced. We conclude that microbial diversity alters bulk chemical structure, the decomposition of plant litter sugars and influences the microbial oxidation of total vanillyl–lignins, thus changing SOM composition.     

Ecosystem N Distribution and δ<Superscript>15</Superscript>N during a Century of Forest Regrowth after Agricultural Abandonment

Abstract Stable isotope ratios of terrestrial ecosystem nitrogen (N) pools reflect internal processes and input–output balances. Disturbance generally increases N cycling and loss, yet few studies have examined ecosystem δ¹⁵N over a disturbance-recovery sequence. We used a chronosequence approach to examine N distribution and δ¹⁵N during forest regrowth after agricultural abandonment. Site ages ranged from 10 to 115 years, with similar soils, climate, land-use history, and overstory vegetation (white pine Pinus strobus). Foliar N and δ¹⁵N decreased as stands aged, consistent with a progressive tightening of the N cycle during forest regrowth on agricultural lands. Over time, foliar δ¹⁵N became more negative, indicating increased fractionation along the mineralization–mycorrhizal–plant uptake pathway. Total ecosystem N was constant across the chronosequence, but substantial internal N redistribution occurred from the mineral soil to plants and litter over 115 years (>25% of ecosystem N or 1,610 kg ha⁻¹). Temporal trends in soil δ¹⁵N generally reflected a redistribution of depleted N from the mineral soil to the developing O horizon. Although plants and soil δ¹⁵N are coupled over millennial time scales of ecosystem development, our observed divergence between plants and soil suggests that they can be uncoupled during the disturbance-regrowth sequence. The approximate 2‰ decrease in ecosystem δ¹⁵N over the century scale suggests significant incorporation of atmospheric N, which was not detected by traditional ecosystem N accounting. Consideration of temporal trends and disturbance legacies can improve our understanding of the influence of broader factors such as climate or N deposition on ecosystem N balances and δ¹⁵N. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Vascular plant 15N natural abundance in heath and forest tundra ecosystems is closely correlated with presence and type of mycorrhizal fungi in roots

In this study we show that the natural abundance of the nitrogen isotope 15, δ¹⁵N, of plants in heath tundra and at the tundra-forest ecocline is closely correlated with the presence and type of mycorrhizal association in the plant roots. A total of 56 vascular plant species, 7 moss species, 2 lichens and 6 species of fungi from four heath and forest tundra sites in Greenland, Siberia and Sweden were analysed for δ¹⁵N and N concentration. Roots of vascular plants were examined for mycorrhizal colonization, and the soil organic matter was analysed for δ¹⁵N, N concentration and soil inorganic, dissolved organic and microbial N. No arbuscular mycorrhizal (AM) colonizations were found although potential host plants were present in all sites. The dominant species were either ectomycorrhizal (ECM) or ericoid mycorrhizal (ERI). The δ¹⁵N of ECM or ERI plants was 3.5–7.7‰ lower than that of non-mycorrhizal (NON) species in three of the four sites. This corresponds to the results in our earlier study of mycorrhiza and plant δ¹⁵N which was limited to one heath and one fellfield in N Sweden. Hence, our data suggest that the δ¹⁵N pattern: NON/AM plants > ECM plants ≥ ERI plants is a general phenomenon in ecosystems with nutrient-deficient organogenic soils. In the fourth site, a␣birch forest with a lush herb/shrub understorey, the differences between functional groups were considerably smaller, and only the ERI species differed (by 1.1‰) from the NON species. Plants of all functional groups from this site had nearly twice the leaf N concentration as that found in the same species at the other three sites. It is likely that low inorganic N availability is a prerequisite for strong δ¹⁵N separation among functional groups. Both ECM roots and fruitbodies were ¹⁵N enriched compared to leaves which suggests that the difference in δ¹⁵N between plants with different kinds of mycorrhiza could be due to isotopic fractionation at the␣fungal-plant interface. However, differences in δ¹⁵N between soil N forms absorbed by the plants could also contribute to the wide differences in plant δ¹⁵N found in most heath and forest tundra ecosystems. We hypothesize that during microbial immobilization of soil ammonium the microbial N pool could become ¹⁵N-depleted and the remaining, plant-available soil ammonium ¹⁵N-enriched. The latter could be a main source of N for NON/AM plants which usually have high δ¹⁵N. In contrast, amino acids and other soil organic N compounds presumably are ¹⁵N-depleted, similar to plant litter, and ECM and ERI plants with high uptake of these N forms hence have low leaf δ¹⁵N. Further indications come from the δ¹⁵N of mosses and lichens which was similar to that of ECM plants. Tundra cryptogams (and ECM and ERI plants) have previously been shown to have higher uptake of amino acid than ammonium N; their low δ¹⁵N might therefore reflect the δ¹⁵N of free amino acids in the soil. The concentration of dissolved organic N was 3–16 times higher than that of inorganic N in the sites. Organic nitrogen could be an important N source for ECM and, in particular, ERI plants in heath and forest tundra ecosystems with low release rate of inorganic N from the soil organic matter. Received: 8 June 1997 / Accepted: 28 February 1998     

Effects of atmospheric CO2 enrichment on δ 13C, δ 15N values and turnover times of soil organic matter pools isolated by thermal techniques

CO₂ applied for Free-Air CO₂ Enrichment (FACE) experiments is strongly depleted in ¹³C and thus provides an opportunity to study C turnover in soil organic matter (SOM) based on its δ ¹³C value. Simultaneous use of ¹⁵N labeled fertilizers allows N turnover to be studied. Various SOM fractionation approaches (fractionation by density, particle size, chemical extractability etc.) have been applied to estimate C and N turnover rates in SOM pools. The thermal stability of SOM coupled with C and N isotopic analyses has never been studied in experiments with FACE. We tested the hypothesis that the mean residence time (MRT) of SOM pools is inversely proportional to its thermal stability. Soil samples from FACE plots under ambient (380 ppm) and elevated CO₂ (540 ppm; for 3 years) treatments were analyzed by thermogravimetry coupled with differential scanning calorimetry (TG-DSC). Based on differential weight losses (TG) and energy release or consumption (DSC), five SOM pools were distinguished. Soil samples were heated up to the respective temperature and the remaining soil was analyzed for δ ¹³C and δ ¹⁵N by IRMS. Energy consumption and mass losses in the temperature range 20–200°C were mainly connected with water volatilization. The maximum weight losses occurred from 200–310°C. This pool contained the largest amount of carbon: 61% of the total soil organic carbon in soil under ambient treatment and 63% in soil under elevated CO₂, respectively. δ ¹³C values of SOM pools under elevated CO₂ treatment showed an increase from −34.3‰ of the pool decomposed between 20–200°C to −18.1‰ above 480°C. The incorporation of new C and N into SOM pools was not inversely proportional to its thermal stability. SOM pools that decomposed between 20–200 and 200–310°C contained 2 and 3% of the new C, with a MRT of 149 and 92 years, respectively. The pool decomposed between 310–400°C contained the largest proportion of new C (22%), with a MRT of 12 years. The amount of fertilizer-derived N after 2 years of application in ambient and elevated CO₂ treatments was not significantly different in SOM pools decomposed up to 480°C having MRT of about 60 years. In contrast, the pool decomposed above 480°C contained only 0.5% of new N, with a MRT of more than 400 years in soils under both treatments. Thus, the separation of SOM based on its thermal stability was not sufficient to reveal pools with contrasting turnover rates of C and N. Responsible Editor: Bernard Nicolardot.     

Box-modeling of 15N/14N in mammals

The ¹⁵N/¹⁴N signature of animal proteins is now commonly used to understand their physiology and quantify the flows of nutrient in trophic webs. These studies assume that animals are predictably ¹⁵N-enriched relative to their food, but the isotopic mechanism which accounts for this enrichment remains unknown. We developed a box model of the nitrogen isotope cycle in mammals in order to predict the ¹⁵N/¹⁴N ratios of body reservoirs as a function of time, N intake and body mass. Results of modeling show that a combination of kinetic isotope fractionation during the N transfer between amines and equilibrium fractionation related to the reversible conversion of N-amine into ammonia is required to account for the well-established ≈4‰ ¹⁵N-enrichment of body proteins relative to the diet. This isotopic enrichment observed in proteins is due to the partial recycling of ¹⁵N-enriched urea and the urinary excretion of a fraction of the strongly ¹⁵N-depleted ammonia reservoir. For a given body mass and diet δ¹⁵N, the isotopic compositions are mainly controlled by the N intake. Increase of the urea turnover combined with a decrease of the N intake lead to calculate a δ¹⁵N increase of the proteins, in agreement with the observed increase of collagen δ¹⁵N of herbivorous animals with aridity. We further show that the low δ¹⁵N collagen values of cave bears cannot be attributed to the dormancy periods as it is commonly thought, but inversely to the hyperphagia behavior. This model highlights the need for experimental investigations performed with large mammals in order to improve our understanding of natural variations of δ¹⁵N collagen.     

Influence of substratum on the variability of benthic biofilm stable isotope signatures: implications for energy flow to a primary consumer

Benthic biofilms have been identified using stable isotope analysis (SIA) as an important resource supporting many freshwater food webs. However, biofilm δ¹³C signatures are highly variable in freshwaters, which may hamper our understanding of energy flow through food webs in these systems. There has been little consideration of the influence that substratum may have on biofilm δ¹³C signature variability and energy flows to primary consumers. We investigated the effect of organic and inorganic substrata on biofilm dynamics by examining: (1) temporal variability of biofilm stable isotope (δ¹³C, δ¹⁵N) signatures on allochthonous leaf-litter (Eucalyptus camaldulensis) and cobble substrata over 12 months in a lowland river in south-eastern Australia; and (2) the effect of substrata on biofilm energy flows to a grazer snail, Physa acuta (Gastropoda: Physidae), using SIA and ecological stoichiometry in a laboratory experiment. The temporal study indicated that cobble biofilm varied significantly in δ¹³C signature during the 12 months (up to 11‰), whereas the δ¹³C signature of leaf biofilm was less variable (less than 2‰). In contrast, biofilm δ¹⁵N signatures varied temporally on both cobble (2.6‰) and leaf (1‰) substrata. This suggests that leaf biofilm was more reliant on leaf tissue for carbon and therefore less limited by carbon supply than cobble biofilm whereas for nitrogen biofilm on both substrata was reliant on external sources. In the laboratory experiment, snails fed leaf biofilm reflected more of an allochthonous δ¹³C signature than cobble biofilm fed snails, suggesting assimilation of leaf carbon via the heterotrophic microbial community within the biofilm. Snails grew largest on cobble biofilm, which had lower C:N ratios than leaf biofilm. Our results demonstrate that the type of substratum can influence the temporal variability of biofilm δ¹³C signatures and energy flow to primary consumers.     

Evaluating the utility of stable isotope analyses of archived freshwater sample materials

We evaluated the potential utility of stable isotope analysis of tissues commonly archived by aquatic biologists. Previous studies with chemically preserved samples have shown contradictory results, which present an obstacle for the use of archived sample materials. We tested the effects of ethanol and formalin preservation on zooplankton and of ethanol on benthic macroinvertebrate δ¹³C and δ¹⁵N values. We found that neither formalin nor ethanol had a significant effect on δ¹³C and δ¹⁵N values of preserved zooplankton. Nor did ethanol significantly affect δ¹³C or δ¹⁵N values of macroinvertebrates. However, ethanol preservation slightly, but significantly decreased C:N ratios of both zooplankton and macroinvertebrates, probably reflecting some extraction of lipids. Overall, the effects of preservatives on δ¹³C and δ¹⁵N values that we observed were minor. We also compared δ¹³C and δ¹⁵N values analysed from roach scales and perch operculum bones with those analysed from muscle tissue. Decalcification of scales and operculum bones only slightly improved our comparison to muscle tissue δ¹³C and δ¹⁵N values. Decalcified scales had slightly higher δ¹³C and lower δ¹⁵N values. Similarly, decalcified operculum bones showed slightly increased δ¹³C and decreased δ¹⁵N values to those for fish muscle. Our results confirm that scales and operculum bones can provide a suitable proxy for fish muscle in isotope studies with minor correction. We conclude that various archived sample materials can indeed be used with confidence for historical reconstructions of freshwater food webs by stable isotope analysis. Handling editor: K. Martens     

Can stable isotope (δ<Superscript>13</Superscript>C and δ<Superscript>15</Superscript>N) measurements of little auk (<Emphasis Type="Italic">Alle alle</Emphasis>) adults and chicks be used to track changes in high-Arctic marine foodwebs?

The little auk (Alle alle), a small and abundant planktivorous seabird that breeds in the high Arctic, has the potential to be used as a monitor of the composition and abundance of lower trophic-level zooplankton. We investigated age- and sex-related sources of variation in diet and stable isotope (δ¹³C and δ¹⁵N) values of little auks breeding in Spitsbergen during the summer of 2002 to evaluate this possibility. Stable isotope profiles of both adult and chick blood changed over the breeding season, with blood δ¹⁵N values increasing and δ¹³C values decreasing. This could represent a switch to higher trophic-level prey derived from more pelagic sources. However, while chick blood δ¹³C values followed those values in their meals, this was not the case for blood δ¹⁵N values, suggesting additional physiological mechanisms influencing blood δ¹⁵N values in growing chicks. Chicks had consistently lower δ¹⁵N values than their parents, which may indicate they were being fed on lower trophic-level prey items or may alternatively reflect complexities in chick blood δ¹⁵N values through the growth period. These results have several important implications for use of stable isotope analysis as a tool to detect changes in seabird diet and availability of lower trophic-level prey in high-Arctic marine environments. Until physiological aspects of stable isotope discrimination are well understood, we caution against using chicks of this seabird as any form of isotopic monitor.     

Order from disorder: do soil organic matter composition and turnover co-vary with iron phase crystallinity?

Soil organic matter (SOM) often increases with the abundance of short-range-ordered iron (SRO Fe) mineral phases at local to global scales, implying a protective role for SRO Fe. However, less is known about how Fe phase composition and crystal order relate to SOM composition and turnover, which could be linked to redox alteration of Fe phases. We tested the hypothesis that the composition and turnover of mineral-associated SOM co-varied with Fe phase crystallinity and abundance across a well-characterized catena in the Luquillo Experimental Forest, Puerto Rico, using dense fractions from 30 A and B horizon soil samples. The δ¹³C and δ¹⁵N values of dense fractions were strongly and positively correlated (R²?=?0.75), indicating microbial transformation of plant residues with lower δ¹³C and δ¹⁵N values. However, comparisons of dense fraction isotope ratios with roots and particulate matter suggested a greater contribution of plant versus microbial biomass to dense fraction SOM in valleys than ridges. Similarly, diffuse reflectance infrared Fourier transform spectroscopy indicated that SOM functional groups varied significantly along the catena. These trends in dense fraction SOM composition, as well as ?¹⁴C values indicative of turnover rates, were significantly related to Fe phase crystallinity and abundance quantified with selective extractions. Mössbauer spectroscopy conducted on independent bulk soil samples indicated that nanoscale ordered Fe oxyhydroxide phases (nano-goethite, ferrihydrite, and/or very-SRO Fe with high substitutions) dominated (66–94%) total Fe at all positions and depths, with minor additional contributions from hematite, silicate and adsorbed Fe^II, and ilmenite. An additional phase that could represent organic-Fe^III complexes or aluminosilicate-bearing Fe^III was most abundant in valley soils (17–26% of total Fe). Overall, dense fraction samples with increasingly disordered Fe phases were significantly associated with increasingly plant-derived and faster-cycling SOM, while samples with relatively more-crystalline Fe phases tended towards slower-cycling SOM with a greater microbial component. Our data suggest that counter to prevailing thought, increased SRO Fe phase abundance in dynamic redox environments could facilitate transient accumulation of litter derivatives while not necessarily promoting long-term C stabilization.     

Changes in stable isotopic signatures of soil nitrogen and carbon during 40 years of forest development

Understanding what governs patterns of soil δ¹⁵N and δ¹³C is limited by the absence of these data assembled throughout the development of individual ecosystems. These patterns are important because stable isotopes of soil organic N and C are integrative indicators of biogeochemical processing of soil organic matter. We examined δ¹⁵N of soil organic matter (δ¹⁵N_SOM) and δ¹³C_SOM of archived soil samples across four decades from four depths of an aggrading forest in southeastern USA. The site supports an old-field pine forest in which the N cycle is affected by former agricultural fertilization, massive accumulation of soil N by aggrading trees over four decades, and small to insignificant fluxes of N via NH₃ volatilization, nitrification, and denitrification. We examine isotopic data and the N and C dynamics of this ecosystem to evaluate mechanisms driving isotopic shifts over time. With forest development, δ¹³C_SOM became depth-dependent. This trend resulted from a decline of ~2‰ in the surficial 15 cm of mineral soil to −26.0‰, due to organic matter inputs from forest vegetation. Deeper layers exhibited relatively little trend in δ¹³C_SOM with time. In contrast, δ¹⁵N_SOM was most dynamic in deeper layers. During the four decades of forest development, the deepest layer (35–60 cm) reached a maximum δ¹⁵N value of 9.1‰, increasing by 7.6‰. The transfer of >800 kg ha⁻¹ of soil organic N into aggrading vegetation and the forest floor and the apparent large proportion of ectomycorrhizal (ECM) fungi in these soils suggest that fractionation via microbial transformations must be the major process changing δ¹⁵N in these soils. Accretion of isotopically enriched compounds derived from microbial cells (i.e., ECM fungi) likely promote isotopic enrichment of soils over time. The work indicates the rapid rate at which ecosystem development can impart δ¹⁵N_SOM and δ¹³C_SOM signatures associated with undisturbed soil profiles.     

Isotopic signature (<Superscript>15</Superscript>N/<Superscript>14</Superscript>N and <Superscript>13</Superscript>C/<Superscript>12</Superscript>C) confirms similarity of trophic niches of millipedes (Myriapoda,diplopoda) in a temperate deciduous forest

The species composition, abundance, and isotopic signature of millipedes (Myriapoda, Diplopoda) were investigated in seven biotopes of Kaluzhskie Zaseki State Nature Reserve. Nine Diplopoda species were found in total, and the local species diversity (within a sampling plot) reached seven species. The Diplopoda tissues were similar to the plant litter in the isotopic composition of nitrogen (δ¹⁵N was by 0.4‰ higher, on average), but were strongly enriched in heavy carbon (δ¹³C was by 4‰ higher, on average). Removal of mineral carbon from the cuticle reduced δ¹³C of Diplopoda by about 1.4‰ on average. Differences in the δ¹⁵N and δ¹³C values between the species did not exceed 2.5‰. Differences in the isotopic compositions of the considered species were small, and, it is impossible to distinguish particular trophic guilds in the Diplopoda community. Analysis of the published data confirmed that isotopic differentiation of millipedes was much less pronounced than in other investigated groups of soil animals. Hence, millipedes of the deciduous forest form a uniform trophic group.     

Large contribution of arbuscular mycorrhizal fungi to soil carbon pools in tropical forest soils

The origins and composition of soil organic matter (SOM) are still largely uncertain. Arbuscular mycorrhizal fungi (AMF) are recognized as indirect contributors through their influence on soil aggregation, plant physiology, and plant community composition. Here we present evidence that AMF can also make large, direct contributions to SOM. Glomalin, a recently discovered glycoprotein produced by AMF hyphae, was detected in tropical soils in concentrations of over 60 mg cm^–3. Along a chronosequence of soils spanning ages from 300 to 4.1 Mio years, a pattern of glomalin concentrations is consistent with the hypothesis that this protein accumulates in soil. Carbon dating of glomalin indicated turnover at time scales of several years to decades, much longer than the turnover of AMF hyphae (which is assumed to be on the order of days to weeks). This suggests that contributions of mycorrhizae to soil carbon storage based on hyphal biomass in soil and roots may be an underestimate. The amount of C and N in glomalin represented a sizeable amount (ca. 4–5%) of total soil C and N in the oldest soils. Our results thus indicate that microbial (fungal) carbon that is not derived from above- or below-ground litter can make a significant contribution to soil carbon and nitrogen pools and can far exceed the contributions of soil microbial biomass (ranging from 0.08 to 0.2% of total C for the oldest soils).     

Tracking the fate of digesta <Superscript>13</Superscript>C and <Superscript>15</Superscript>N compositions along the ruminant gastrointestinal tract: Does digestion influence the relationship between diet and faeces?

Faecal stable isotope compositions reflect wildlife diets, if digestive processes along the gastrointestinal tract (GIT) do not alter diet–faeces isotopic relationships in an unpredictable way. We investigated ¹³C and ¹⁵N compositions of digesta along the ruminant GIT, using Saanen dairy goats kept on pure grass hay or browse for >20 days. Isotopic changes occurred in the ventral rumen, and in the small intestine, where digesta had significantly higher δ¹³C and δ¹⁵N (associated with lower C or higher N content, respectively) values relative to other GIT sites. However, effects on isotope fractionation were small (∼1.0‰ for δ¹³C and ∼ 2.0‰ for δ¹⁵N), and were reversed in the hindgut such that faecal isotope compositions did not differ from the foregut. No other substantial isotopic changes occurred across GIT sites, despite the morphophysiological complexity of the ruminant GIT. We found similarly small differences across GIT components of rheem gazelles (Gazella leptoceros) fed a mixture of C₃ lucerne and C₄ grass, although in this case faeces were ¹⁵N-depleted relative to other GIT components. Along with differences in δ¹⁵N between goats fed browse or grass, this result implies a systematic difference in diet–faeces δ¹⁵N relationships, contingent on the botanical composition of ruminant diets. Thus, while our results support faecal δ¹³C as a reliable proxy for wildlife diets, further work on factors influencing faecal ¹⁵N abundance is needed. Finally, we note high levels of isotopic variability between individuals fed the same diets, even accounting for the relatively short duration of the experiments, suggesting an important influence of stochasticity on isotope fractionation.     

Effects of nutritional restriction on nitrogen and carbon stable isotopes in growing seabirds

When using stable isotopes as dietary tracers it is essential to consider effects of nutritional state on isotopic fractionation. While starvation is known to induce enrichment of ¹⁵N in body tissues, effects of moderate food restriction on isotope signatures have rarely been tested. We conducted two experiments to investigate effects of a 50–55% reduction in food intake on δ¹⁵N and δ¹³C values in blood cells and whole blood of tufted puffin chicks, a species that exhibits a variety of adaptive responses to nutritional deficits. We found that blood from puffin chicks fed ad libitum became enriched in ¹⁵N and ¹³C compared to food-restricted chicks. Our results show that ¹⁵N enrichment is not always associated with food deprivation and argue effects of growth on diet–tissue fractionation of nitrogen stable isotopes (Δ¹⁵N) need to be considered in stable isotope studies. The decrease in δ¹³C of whole blood and blood cells in restricted birds is likely due to incorporation of carbon from ¹³C-depleted lipids into proteins. Effects of nutritional restriction on δ¹⁵N and δ¹³C values were relatively small in both experiments (δ¹⁵N: 0.77 and 0.41‰, δ¹³C: 0.20 and 0.25‰) compared to effects of ecological processes, indicating physiological effects do not preclude the use of carbon and nitrogen stable isotopes in studies of seabird ecology. Nevertheless, our results demonstrate that physiological processes affect nitrogen and carbon stable isotopes in growing birds and we caution isotope ecologists to consider these effects to avoid drawing spurious conclusions.     

Carbon isotopes reveal soil organic matter dynamics following arid land shrub expansion

 Over the past century, overgrazing and drought in New Mexico’s Jornada Basin has promoted the replacement of native black grama (Bouteloua eriopoda Torr.) grass communities by shrubs, primarily mesquite (Prosopis glandulosa Torr.). We investigated the effects of shrub expansion on the distribution, origin, turnover, and quality of light (LFC) and heavy (HFC) soil organic matter (SOM) fractions using δ¹³C natural abundance to partition SOM into C₄ (grass) and C₃ (shrub) sources. Soil organic matter beneath grasses and mesquite was isotopically distinct from associated plant litter, providing evidence of both recent shrub expansion and Holocene plant community changes. Our δ¹³C analyses indicated that SOM derived from mesquite was greatest beneath shrub canopies, but extended at least 3 m beyond canopy margins, similar to the distribution of fine roots. Specific ¹⁴C activities of LFC indicated that root litter is an important source of SOM at depth. Comparison of turnover rates for surface LFC pools in grass (7 or 40 years) and mesquite (11 or 28 years) soils and for HFC pools by soil depth (∼150–280 years), suggest that mesquite may enhance soil C storage relative to grasses. We conclude that the replacement of semiarid grasslands by woody shrubs will effect changes in root biomass, litter production, and SOM cycling that influence nutrient availability and long-term soil C sequestration at the ecosystem level. Received: 17 May 1996 / Accepted: 12 November 1996