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The long-distance migrations by marine fishes are difficult to track by field observation. Here, we propose a new method to track such migrations using stable nitrogen isotopic composition at the base of the food web (δ15 NBase ), which can be estimated by using compound-specific isotope analysis. δ15 NBase exclusively reflects the δ15 N of nitrate in the ocean at a regional scale and is not affected by the trophic position of sampled organisms. In other words, δ15 NBase allows for direct comparison of isotope ratios between proxy organisms of the isoscape and the target migratory animal. We initially constructed a δ15 NBase isoscape in the northern North Pacific by bulk and compound-specific isotope analyses of copepods (n = 360 and 24, respectively), and then we determined retrospective δ15 NBase values of spawning chum salmon (Oncorhynchus keta) from their vertebral centra (10 sections from each of two salmon). We then estimated the migration routes of chum salmon during their skeletal growth by using a state-space model. Our isotope tracking method successfully reproduced a known chum salmon migration route between the Okhotsk and Bering seas, and our findings suggest the presence of a new migration route to the Bering Sea Shelf during a later growth stage.

Dissolved organic carbon (DOC) plays a major role in aquatic carbon cycling. In order to determine if DOC photolability was related to DOC age, we tested the effect of oxidation time using UV irradiation on natural radiocarbon concentrations (Δ14C) of DOC in six US rivers. The most photo-labile DOC was 14C youngest. After a 100 min UV irradiation, most DOC (>93%) was oxidized in two out of the six rivers studied. On the other hand, the other four rivers still had a considerable amount (>20%) of DOC resistant to UV oxidation, which showed lower Δ14C values than those of bulk DOC. We also found that the concentration and Δ14C value of bulk DOC are controlled by the photochemically intermediate fraction. The results suggest that 14C age of organic matter is not homogeneous within DOC, the degree of which may depend on the characteristics unique to rivers.

Rationale To achieve better precision and accuracy for δ13 C analysis of individual amino acids (AAs), we have developed a new analytical method based on multi-dimensional high-performance liquid chromatography (HPLC) and elemental analyzer/isotope ratio mass spectrometry (EA/IRMS). Unlike conventional methods using gas chromatography, this approach omits pre-column chemical derivatization, thus reducing systematic errors associated with the isotopic measurement. Methods The separation and isolation of individual AAs in a standard mixture containing 15 AAs and a biological sample, spear squid (Heterololigo bleekeri) were performed. AAs were isolated using an HPLC system equipped with a reversed-phase column and a mixed-mode column and collected using a fraction collector. After the chromatographic separation and further post-HPLC purification, the δ13 C values of AAs were measured by EA/IRMS. Results The complete isolation of all 15 AAs in the standard mixture was achieved. The δ13 C values of these AAs before and after the experiment were in good agreement. Also, 15 AAs in the biological sample, H. bleekeri, were successfully measured. The δ13 C values of AAs in H. bleekeri varied by as much as 30‰ with glycine being most enriched in13 C. Conclusions The consistency between the δ13 C values of reference and processed AAs demonstrates that the experimental procedure generates accurate δ13 C values unaffected by fractionation effects and contamination. This method is therefore suitable for δ13 C analysis of biological samples with higher precision than conventional approaches. We propose this new method as a tool to measure δ13 C values of AAs in biological, ecological and biogeochemical studies.

We have developed an analytical method for the precise δ13C measurement of individual amino acid using a multi-dimensional high-performance liquid chromatography (HPLC) and a nano-scale elemental analyzer/isotope ratio mass spectrometry (EA/IRMS). Although this method is time-consuming, it can offer higher precision and accuracy than does the conventional analytical method such as GC/C/IRMS, because the derivatization of amino acids is not required. A reversed-phase column (CAPCELL PAK C18, Shiseido, Japan) and a mixed-mode column (Primesep A, SIELC Technologies, U.S.A.) were applied for the HPLC (Agilent Technologies, U.S.A.) with a charged aerosol detector (Thermo Fisher Scientific, U.S.A.) (Ishikawa et al., 2018). We conducted the isolation of underivatized amino acids in a standard mixture containing 15 proteinogenic amino acids (Gly, Ala, Glu, Arg, Val, Pro, Met, Tyr, Ile, Leu, Phe, Thr, His, Asp, Ser), and confirmed that all these amino acids were successfully isolated. Each collected amino acid was filtered through a 0.45 μm membrane filter (Pall, U.S.A.) and washed with diethyl ether to remove hydrophobic impurities. The δ13C values of these amino acids before and after the separation and purification were consistent, which proved that the whole experimental procedure did not change the δ13C values of amino acids. We applied this method to several aquatic organisms. The results show that the δ13C values of amino acids vary as large as 30‰ with Gly being most enriched in 13C.ReferenceIshikawa et al., (2018) Anal. Chem., 90, 20, 12035-12041.

Tracing of biogeochemical pathways using molecular approaches has advanced our basic understanding of the carbon cycle and life’s legacy in the sedimentary record. To this end, compound-specific radiocarbon analysis has been instrumental in shedding light on the turnover, age, and sources of a range of biomarkers embedded within complex environmental matrices. However, despite their foundational importance for life and their omnipresence throughout geologic space and time, the biogeochemical cycling of amino acids remains largely unexplored. Here, we discuss the potential of using amino acid-specific radiocarbon to deepen our knowledge of the biogeochemistry of food webs and sedimentary organic carbon., Frontiers in Marine Science, 7, ISSN:2296-7745

The catchment of Lake Biwa, the largest freshwater lake in Japan, has experienced rapid urbanization and industrialization in the second half of the 20th century; yet little is known about the historical trend of metal pollution. We analyzed the elemental concentrations and zinc stable isotope ratios (δ66Zn) in a sediment core from the Northern basin of the lake. The goals were 1) to reconstruct the historical trend of trace metals in the lake and 2) to understand whether the range of Zn concentrations was derived from natural or anthropogenic sources. The first increase in Cu and Zn concentrations from the late 1910s until approximately 1940 could be due to human-induced higher contributions from the surrounding mafic igneous rocks. A second increase in the Zn concentrations accompanied by a slight decrease in the δ66Zn values from the late 1950s coincided with the period of the industrialization and urbanization. The δ66Zn values fell in a binary mixing process between a lithogenic (∼+0.28‰) and an anthropogenic endmember (∼+0.14‰). Although industrial effluents were strictly regulated by law from the late 1960s, constant Zn concentrations throughout the 1980s indicated contributions from domestic wastewaters and diffusive sources such as runoff from paddy fields, roads, and industrial areas. Zinc, Ni, and Co were still enriched in recent surface sediment compared to the pre-industrial times. The metal fractions obtained by the BCR sequential extraction procedure showed that Pb was mainly hosted in the reducible fraction, and Cu and Zn were mainly hosted in the residual fraction. The increasing trace metal concentrations in the acid-labile and reducible fractions towards the surface indicate these fractions were the primary host for anthropogenic trace metals. We conclude it is unlikely that the natural condition of the lake sediment will be recovered owing to the continuing urbanization around Lake Biwa.

The trophic discrimination factor (TDF) of nitrogen isotopes (15N/14N) within amino acids, between a stream-dwelling dobsonfly larva (Protohermes grandis: Megaloptera; Corydalidae) and its diet (chironomid larvae), was determined in controlled feeding experiments. Last-instar larvae of P. grandis were collected from the Yozawa-gawa River, central Japan, and reared in the laboratory. After fed to satiation for 1 month, one group of larvae was each fed one living chironomid larva per day for 4 weeks, while a second group was starved for 8 weeks. The larvae were harvested at intervals and the nitrogen isotopic composition of glutamic acid (δ15NGlu) and phenylalanine (δ15NPhe) were determined to calculate TDF. The mean TDF of satiated and starved larvae were 7.1‰ ± 0.5‰ (n = 3) and 7.3‰ ± 0.5‰ (n = 5), respectively. Thus, the TDF for P. grandis larvae in this study was similar to that reported for other arthropods (approximately 7‰) and was independent of satiation or starvation. A previous study of wild P. grandis larvae, based on the δ15NGlu and δ15NPhe values, estimated its trophic position (TP) as approximately 2.0 ± 0.1 (n = 5), a low value close to that of algivores, although they are generally characterized as carnivores (usually accepted as TP ≥ 3). The TDF for P. grandis larvae suggests that their low TPs in nature were caused by incorporation of vascular plant-derived amino acids (with a different δ15N profile from that of algae) and not by an unusually low TDF or by the effects of the satiation/starvation on amino acid metabolism.

We examine instrumental and methodological capabilities for microscale (10-50 μg of C) radiocarbon analysis of individual compounds in the context of paleoclimate and paleoceanography applications, for which relatively high-precision measurements are required. An extensive suite of data for

We have improved a method for isolation and purification of individual amino acids for compound-specific radiocarbon analysis (CSRA). To remove high-performance liquid chromatography (HPLC) eluent blanks from isolated amino acid fractions prior to the radiocarbon (Δ14C) measurement, each fraction was filtered through a membrane filter and then washed with diethyl ether twice. Radiocarbon measurements on standard amino acids processed and purified with the above method using elemental analyzer–accelerator mass spectrometry resulted in Δ14C values that were in strong agreement (R2 = 0.998) with the original Δ14C value of each amino acid standard. From these measurements, we calculate dead and modern carbon contamination contributions as 1.2 ± 0.2 and 0.3 ± 0.1 μgC, respectively, which are consistent with direct assessments of HPLC procedural blanks of 1.0 ± 0.8 μgC per sample. These contamination constraints allow correction of measured Δ14C values for accurate and precise CSRA and are widely applicable to ...

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., Ecological Research, 33 (5), ISSN:0912-3814

Integration of inland waters into regional and global carbon (C) budgets requires a comprehensive understanding of factors regulating organic carbon (OC) delivery and in situ processing. This study reviews advances in optical, molecular, and isotopic approaches to resolve the sources, ages, and transformations of OC in aquatic systems. OC characterization using excitation emission matrix spectra, Fourier transform ion cyclotron mass spectrometry, and nuclear magnetic resonance provides detailed molecular level insight. Radiocarbon isotopic approaches and compound‐specific techniques resolve the input, metabolic fate, and turnover time of OC in ecosystems ranging in size from streams to the open ocean. Accumulating evidence suggests that aquatic OC is composed of diverse biogeochemical components. We conclude with enduring and emerging questions that underscore the role of inland systems in the global C cycle and propose unique combinations of approaches to better discern their role in the delivery and transformation of OC from soils to seas.

Periphytic algae attached to a streambed substrate (periphyton) are an important primary producer in stream ecosystems. We determined the isotopic composition of chlorophyll a in periphyton collected from a stream flowing on limestone bedrock in the Seri River, central Japan. Stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) and natural radiocarbon abundances (Δ14C) were measured in chlorophyll a (δ13Cchl, δ15Nchl and Δ14Cchl) and bulk (δ13Cbulk, δ15Nbulk and Δ14Cbulk) for periphyton, a pure aquatic primary producer (Cladophora sp.) and a terrestrial primary producer (Quercus glauca). Periphyton δ13Cbulk and δ13Cchl values did not necessarily correspond to δ13Cbulk for an algal-grazing specialist (Epeorus latifolium). Periphyton Δ14Cchl values (−258 ‰ in April and −190 ‰ in October) were slightly lower than Δ14Cbulk values (−228 ‰ in April and −179 ‰ in October) but were close to the Δ14C value for dissolved inorganic carbon (DIC; −217 ± 31 ‰), which is a mixture of weathered carbonates (Δ14C = −1000 ‰), CO2 derived from aquatic and terrestrial organic matters (variable Δ14C) and dissolved atmospheric CO2 (Δ14C approximately +30 ‰ in 2013). Δ14Cchl values were also close to Δ14Cbulk for E. latifolium (−215 ‰ in April and −199 ‰ in October) and Cladophora sp. (−210 ‰), whereas the Δ14Cbulk value for Q. glauca (+27 ‰) was closer to Δ14C for atmospheric CO2. Although the bulk isotopic composition of periphyton is recognised as a surrogate for the photosynthetic algal community, natural periphyton is a mixture of aquatic and terrestrial organic materials. Our results indicate that the bulk periphyton matrix at the study site consists of 89 to 95 % algal carbon (derived from 14C-depleted DIC) and 5 to 11 % terrestrial organic carbon (derived from 14C-enriched atmospheric CO2).

Food-web complexity often hinders disentangling functionally relevant aspects of food-web structure and its relationships to biodiversity. Here, we present a theoretical framework to evaluate food-web complexity in terms of biodiversity. Food network unfolding is a theoretical method to transform a complex food web into a linear food chain based on ecosystem processes. Based on this method, we can define three biodiversity indices, horizontal diversity (DH ), vertical diversity (DV ) and range diversity (DR ), which are associated with the species diversity within each trophic level, diversity of trophic levels, and diversity in resource use, respectively. These indices are related to Shannon's diversity index (H'), where H' = DH + DV - DR . Application of the framework to three riverine macroinvertebrate communities revealed that D indices, calculated from biomass and stable isotope features, captured well the anthropogenic, seasonal, or other within-site changes in food-web structures that could not be captured with H' alone.

The relationship between biodiversity and ecosystem functioning is an important theme in environmental sciences. We propose a new index for configuration of the biomass pyramid in an ecosystem, named integrated trophic position (iTP). The iTP is defined as a sum of trophic positions (i.e. the average number of steps involved in biomass transfer) of all the animals in a food web integrated by their individual biomass. The observed iTP for stream macroinvertebrates ranged from 2.39 to 2.79 and was negatively correlated with the species density and the Shannon–Wiener diversity index of the local community. The results indicate a lower efficiency of biomass transfer in more diverse communities, which may be explained by the variance in edibility hypothesis and/or the trophic omnivory hypothesis. We found a negative effect of biodiversity on ecosystem functioning., Scientific Reports, 7, ISSN:2045-2322

Food webs are known to have myriad trophic links between resource and consumer species. While herbivores have well-understood trophic tendencies, the difficulties associated with characterizing the trophic positions of higher-order consumers have remained a major problem in food web ecology. To better understand trophic linkages in food webs, analysis of the stable nitrogen isotopic composition of amino acids has been introduced as a potential means of providing accurate trophic position estimates. In the present study, we employ this method to estimate the trophic positions of 200 free-roaming organisms, representing 39 species in coastal marine (a stony shore) and 38 species in terrestrial (a fruit farm) environments. Based on the trophic positions from the isotopic composition of amino acids, we are able to resolve the trophic structure of these complex food webs. Our approach reveals a high degree of trophic omnivory (i.e., noninteger trophic positions) among carnivorous species such as marine fish and terrestrial hornets.This information not only clarifies the trophic tendencies of species within their respective communities, but also suggests that trophic omnivory may be common in these webs.

We measured the natural abundances of radiocarbon (delta14C) in macroinvertebrates, fishes, and their potential food sources, collected from the upper and lower reaches of six temperate streams in Lake Biwa basin (central Japan), three of which flow on limestone bedrock. Several carbon storage reservoirs in the watersheds show distinctive delta14C signatures (e.g., ancient carbonate rocks, -1000 per thousand; modern atmospheric CO2, +50 per thousand). Our analyses showed that the delta14C values for periphytic algae range from -361 per thousand to +21 per thousand, reflecting 14C-depleted signals from watershed storage reservoirs (carbonate rocks and/or soils). In contrast, the delta14C values for coarse particulate organic matter (CPOM) range from -6 per thousand to +62 per thousand, reflecting 14C-enriched signals from modern atmospheric CO2. The periphyton from streams on limestone bedrock was more 14C-depleted than that from streams in non-limestone areas, although the delta14C values for periphyton from the latter were less than modern atmospheric 14CO2 concentration. The delta14C values for most of the consumers were between those for periphyton and CPOM. Based on a delta14C two-source mixing models, the results suggested that the grazers rely on periphyton, while the carbon source for collectors and predators shifts from CPOM in the upper reaches of streams to periphyton in the lower reaches. The delta14C signature can trace carbon from watershed storage reservoirs to benthic production, which suggests that stream food webs are composed of mixtures of carbon originating from various sources of different ages.

The stable N isotopic composition of individual amino acids (SIAA) has recently been used to estimate trophic positions (TPs) of animals in several simple food chain systems. However, it is unknown whether the SIAA is applicable to more complex food web analysis. In this study we measured the SIAA of stream macroinvertebrates, fishes, and their potential food sources (periphyton and leaf litter of terrestrial C3 plants) collected from upper and lower sites in two streams having contrasting riparian landscapes. The stable N isotope ratios of glutamic acid and phenylalanine confirmed that for primary producers (periphyton and C3 litter) the TP was 1, and for primary consumers (e.g., mayfly and caddisfly larvae) it was 2. We built a two-source mixing model to estimate the relative contributions of aquatic and terrestrial sources to secondary and higher consumers (e.g., stonefly larva and fishes) prior to the TP calculation. The estimated TPs (2.3-3.5) roughly corresponded to their omnivorous and carnivorous feeding habits, respectively. We found that the SIAA method offers substantial advantages over traditional bulk method for food web analysis because it defines the food web structure based on the metabolic pathway of amino groups, and can be used to estimate food web structure under conditions where the bulk method cannot be used. Our result provides evidence that the SIAA method is applicable to the analysis of complex food webs, where heterogeneous resources are mixed.

Little is known about differences in species diversity among ecological communities subject to different levels of human-caused habitat transformation and how this disturbance contributes to diversity through symbiotic dependencies with the environment in freshwater ecosystems. We estimated α and β diversities of benthic macroinvertebrates and relationships between diversity and environmental variables in Ado River (natural) and Yasu River (intermediately disturbed) watersheds, Japan. Alpha diversity was consistently slightly higher in the natural river watershed than in the intermediately disturbed one, but the spatial distribution was not equivalent. The opposite pattern was found for β diversity. Significant differences in environmental variables existed between the two river watersheds, with especially high chlorophyll-a concentrations detected in the intermediately disturbed watershed. Alpha diversity was not correlated with specific environmental variables, whereas water temperature and chlorophyll-a concentrations were the two most significant environmental variables influencing β diversity across sites in the two watersheds. These results suggest that diversity patterns in freshwater benthic macroinvertebrates are differentially influenced by levels of human-caused habitat transformation, especially that intermediately disturbed habitats may benefit species turnover, and further understanding how they relate to environmental variables is essential for protecting local to regional diversity and can provide useful information for conservation planning to maximise biodiversity at the watershed scale.

We review the use of stable carbon isotope ratios (δ13C) and radiocarbon natural abundances (Δ14C) for stream food web studies. The δ13C value of primary producers (e.g., periphytic algae, hereafter periphyton) in streams is controlled by isotopic fractionation during photosynthesis and variable δ13C of dissolved CO2. When periphyton δ13C differs from that of terrestrial primary producers, the relative contribution of autochthony and allochthony to stream food webs can be calculated. Moreover, the variation in periphyton δ13C can reveal how much stream consumers rely on local resources because each stream habitat (e.g., riffle vs. pool, open vs. shaded) usually has a distinctive δ13C. However, periphyton δ13C often overlaps with that of terrestrial organic matter. On the other hand, periphyton Δ14C is less variable than δ13C among habitats, and reflects the Δ14C of dissolved CO2, which could be a mixture of “aged” (Δ14C 0 ‰) carbon. This is because the Δ14C is corrected by its δ13C value for the isotopic fractionation during photosynthesis. Recent studies and our data indicate that many stream food webs are supported by “aged” carbon derived from the watershed via autochthonous production. The combined use of δ13C and Δ14C allows robust estimation of the carbon transfer pathway in a stream food web at multiple spatial scales ranging from the stream habitat level (e.g., riffle and pool) to watershed level (autochthony and allochthony). Furthermore, the Δ14C of stream food webs will expand our understanding about the time frame of carbon cycles in the watersheds.

The trophic discrimination factor (TDF) of nitrogen isotopes (15N/14N) within amino acids, between a stream‐dwelling dobsonfly larva (Protohermes grandis: Megaloptera; Corydalidae) and its diet (chironomid larvae), was determined in controlled feeding experiments. Last‐instar larvae of P. grandis were collected from the Yozawa‐gawa River, central Japan, and reared in the laboratory. After fed to satiation for 1 month, one group of larvae was each fed one living chironomid larva per day for 4 weeks, while a second group was starved for 8 weeks. The larvae were harvested at intervals and the nitrogen isotopic composition of glutamic acid (δ 15NGlu) and phenylalanine (δ 15NPhe) were determined to calculate TDF. The mean TDF of satiated and starved larvae were 7.1‰ ± 0.5‰ (n = 3) and 7.3‰ ± 0.5‰ (n = 5), respectively. Thus, the TDF for P. grandis larvae in this study was similar to that reported for other arthropods (approximately 7‰) and was independent of satiation or starvation. A previous study of wild P. grandis larvae, based on the δ 15NGlu and δ 15NPhe values, estimated its trophic position (TP) as approximately 2.0 ± 0.1 (n = 5), a low value close to that of algivores, although they are generally characterized as carnivores (usually accepted as TP ≥ 3). The TDF for P. grandis larvae suggests that their low TPs in nature were caused by incorporation of vascular plant‐derived amino acids (with a different δ 15N profile from that of algae) and not by an unusually low TDF or by the effects of the satiation/starvation on amino acid metabolism.

Carbon and nitrogen stable isotope ratios (δ13C and δ15N) have been widely employed in food web analysis. In lotic environments, periphyton is a major primary producer that makes a large contribution to food web production as well as carbon and nitrogen cycling. While the δ13C and δ15N values have many advantages as a natural tracer, the controls over their high spatial and temporal variability in stream periphyton are not well known. Here, we present the global dataset of δ13C and δ15N values of lotic periphyton from 54 published and two unpublished sources, including 978 observations from 148 streams/rivers in 38 regions around the world, from arctic to tropical sites. The 54 published sources were articles recorded during the period of 1994–2016 in 25 academic journals. The two unpublished sources were from the authors’ own data. The dataset showed that δ13C and δ15N values of periphyton ranged from −47.3 to −9.3‰ and from −5.6 to + 22.6‰, respectively. The dataset also includes physicochemical factors (altitude, coordinates, catchment area, width, depth, geology, vegetation, canopy coverage, biome, season, presence of anadromous salmon, temperature, pH, current velocity, and discharge), nutrient data (nitrate and ammonium concentrations), and algal attributes (chlorophyll a concentration, algal species compositions, and carbonates removal) in streams/rivers studied, all of which may help interpret the δ13C and δ15N values of periphyton. The metadata file outlines structure of all the data and with references for data sources, providing a resource for future food web studies in stream and river ecosystems.

We measured radiocarbon concentrations (Δ14C), together with stable isotope ratios (δ13C and δ15N) of aquatic animals, periphytic algae and particulate organic matter (POM) at upstream sites of two streams. We hypothesized that Δ14C values of organisms would be separated into two major carbon sources in streams: in-stream production (autochthonous production) and terrestrial detritus (allochthonous production). In fact, Δ14C values of in-stream production were low, while those of terrestrial detritus were high. This indicates the difference in sources: the former reflects the Δ14C values of the dissolved inorganic carbon in the catchment, whereas the latter reflects the Δ14C value of present atmospheric CO2. Δ14C patterns of animals differed in each of the two streams, suggesting that the role of food web structure is critical for understanding the carbon dynamics of stream ecosystems. We present, for the first time, the potential use of radiocarbon (14C) in the study of food webs in stream ecosystems.

Carbon stable isotope ratios (δ(13)C) are widely used to trace resource transfer pathways, yet δ(13)C variation in freshwater autotrophs is not yet fully understood. We have analyzed data from 42 published studies, supplemented with some unpublished data, to show the determinants of lotic periphyton δ(13)C. At large spatial scales, we observed broad differences in periphyton δ(13)C among biomes and consistent longitudinal variation related to watershed area. Longitudinal increases in δ(13)C indicate the importance of in-stream processes on lotic carbon cycles and autotroph δ(13)C variation. At local spatial scales, periphyton δ(13)C was negatively related to canopy cover and water current velocity and positively related to chlorophyll a density. Autotroph δ(13)C varied among taxonomic groups. Cyanobacteria and red algae had significantly higher and lower δ(13)C than other taxa, respectively. A hierarchical model across spatial scales showed that local controls for periphyton δ(13)C were nested by regional controls, which suggested that productivity and CO(2) availability determine δ(13)C. Overall, our results reveal general patterns of periphyton δ(13)C and provide improved information for study design and the use of δ(13)C in isotopic mixing models in lotic food web studies.

In stream food-web analysis, the contributions of carbon from periphyton (an autochthonous source) and terrestrial litter (an allochthonous source) are estimated by isotopic difference. We hypothesised that periphyton δ13C varies among stream habitats, whereas Δ14C does not because Δ14C is corrected with δ13C, by definition. To test this hypothesis, we compared the variability of δ13C and Δ14C of periphyton and dissolved inorganic carbon (DIC) in four habitats (open v. shaded, riffle v. pool) within a limestone-based upland stream in Japan. Periphyton δ13C values (from –31.9‰ to –16.3‰) were significantly different among the habitats whereas the Δ14C values (from –379‰ to –141‰) were not. Periphyton δ13C values depended on both algal photosynthetic activity and δ13C of the DIC, whereas periphyton Δ14C depended only on DIC Δ14C. The δ13C and Δ14C values of litter were constant. Thus, the difference between the periphyton and litter δ13C values (Δδ13C) varied among habitats, but their Δ14C differences (ΔΔ14C) did not. Our results indicate that Δ14C is spatially stable among stream habitats and that Δ14C measurements can be used to precisely determine carbon sources for stream food-web analysis within individual reaches.

Long-term monitoring of ecosystem succession provides baseline data for conservation and management, as well as for understanding the dynamics of underlying biogeochemical processes. We examined the effects of deforestation and subsequent afforestation of a riparian forest of Japanese cedar (Cryptomeria japonica) on stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) and natural abundances of radiocarbon (Δ14C) in stream biota in the Mt. Gomadan Experimental Forest and the Wakayama Forest Research Station, Kyoto University, central Japan. Macroinvertebrates, periphytic algae attached to rock surfaces (periphyton), and leaf litter of terrestrial plants were collected from six headwater streams with similar climate, topography, and bedrock geology, except for the stand ages of riparian forests (from 3 to 49 yr old in five stands and >90 yr old in one reference stand). Light intensity and δ13C values of both periphyton and macroinvertebrates decreased synchronously with forest age in winter. A Bayesian mixing model indicates that periphyton contributions to the stream food webs are maximized in 23-yr-old forests. Except for grazers, most macroinvertebrates showed Δ14C values similar to those of terrestrial leaf litter, reflecting the influence of modern atmospheric CO2 Δ14C values. On the other hand, the Δ14C values of both periphyton and grazers (i.e., aquatic primary consumers) were significantly lower than that of modern atmospheric CO2, and were lowest in 23-yr -old forest stands. Previous studies show that root biomass of C. japonica peaks at 15–30 yr after planting. These evidences suggest that soil CO2 released by root respiration and dispersed by groundwater weathers carbonate substrata, and that dissolved inorganic carbon (DIC) with low Δ14C is incorporated into stream periphyton and some macroinvertebrates. The ecological response in the studied streams to clear-cutting and replanting of Japanese cedar is much slower (~20 yr) than the chemical response (