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International audience; Peat deposits (>50 ka) in the montane Nilgiris (Western Ghats, India), have been central to the reconstruction of late Quaternary paleoclimate using paleovegetation changes in the forest-grassland vegetation mosaic that coexist here. However, it is well-known that short-term disturbances can also cause vegetation switches when multiple stable vegetation states exist. We studied paleovegetation changes within the alternative stable states framework using stable carbon isotopes (relative abundance of C3-C4 vegetation) on the cellulose fraction from two high-resolution radiocarbon-dated peat cores ~170 m apart in the Sandynallah valley: Core 1 closer to the hillslope (32,000 years old) and Core 2 from the centre of the valley (45,000 years old). Core 1 is located in an ecotone showing shola-sedgeland dynamics with vegetation switching at c.22 ka from shola (possibly due to fire) to a prolonged unstable state until 13 ka sustained by low waterlogging. Following a hiatus c.13 ka, sedgeland dominates, with a shift into shola at 3.75 ka driven by increasing aridity. Core 2 shows a stable sedgeland mixed C3-C4 composition responding to temperature, enriched in C3-vegetation in the last glacial with C4-dominance beginning c.18.5 ka, indicative of deglacial warming. The distinctive vegetation states at corresponding times in Cores 1 and 2 within the same valley, responding independently to disturbances and climate, respectively, is the first paleo-record from an alternative stable states landscape in the montane tropics. Thus, short-term disturbances and site attributes need to be accounted for before ascribing vegetation change to changing climate in such vegetation mosaics.

Ecologists have proposed that montane grassland-shola (stunted evergreen forest) mosaics in the Western Ghats may represent alternative stable vegetation states. But paleoecology investigations sel...

The field diet of meroplanktonic decapod crustacean larvae is poorly known, despite standard use of microzooplankton as food in laboratory culture. Using callianassid shrimp Nihonotrypaea harmandi larvae collected from a 65 m deep inner-shelf location off mid-western Kyushu, Japan, between June and August 2012 and 2013 and mass-reared in the laboratory, a phytoplankton-based diet through larval development (zoeae I?VI to decapodid) was demonstrated. When the pure-cultured diatom Chaetoceros gracilis was fed to zoeae, survival rate to decapodids was 3.4 to 3.9% in 26 to 40 d at 22°C, which was comparable to previous rearing results for zoeae fed microzooplankton. Trophic enrichment factors (TEFs) from stable isotope (SI) analysis of zoeal whole-body tissue in the laboratory were 2.0‰ for δ13C and 1.9‰ for δ15N. In the field water column, diatoms dominated the nano- to micro-sized plankton, accounting for 38 to 81% of the biovolume, followed by heterotrophic protists. The trophic position (TP) estimated from amino acid-specific δ15N values for the field-collected zoeae VI was 2.1 (TPGlu/Phe) or 2.7 (TPAla/Phe), suggesting that those zoeae fed on mixtures of phytoplankton and heterotrophs including protists. Bulk SI analyses were performed for particulate organic matter (POM; proxy for phytoplankton), microzooplankton (mainly calanoid copepods), and shrimp zoeae to elucidate the diet of larvae in the water column. A shift in SI from fresh to degraded POM was determined through the incubation of field-collected POM. Based on this shift during degradation and larval TEFs, phytoplankton and their sinking detritus with heterotrophic protists were estimated to be the principal diet for those larvae residing mostly below the chlorophyll maximum layer., Marine Ecology Progress Series, 592, pp.141-158; 2018

Long term monitoring of atmospheric wet and dry depositions and associated nutrients fluxes was conducted on the coast of Japan facing the East China Sea continuously for 1 year and 2 months, with the origin of air mass investigated based on isotope analyses (Sr, Nd, and NO3). During the same period, intensive observations of ocean conditions and the chemical composition of sinking particles collected using sediment traps were conducted to investigate the effects of atmospheric deposition-derived nutrients on phytoplankton blooms. Dry-deposition-derived nutrient inputs to the surface ocean were larger during autumn to spring than in summer due to the effect of continental air mass occasionally carrying Asian dust (yellow sand). However, these nutrients fluxes were limited (1.1–1.5 mg-N m−2 day−1 on average) and didn't appear to cause phytoplankton blooms through the year. Although average dissolved inorganic nitrogen (DIN) concentrations in rainwater were lower in oceanic air masses compared to continental air masses, wet-deposition-derived nutrient inputs to the surface ocean on rainy days during the summer (26.0 mg-N m−2 day−1 on average) were large due to higher precipitation from oceanic air masses. Wet-deposition-derived nutrients significantly increased nutrient concentrations in the surface ocean and seemed to cause phytoplankton blooms in the warm rainy season when nutrients in the surface were depleted due to increased stratification. The increase in phytoplankton biomass was reflected in increased particle sinking into the bottom layer, as well as changing chemical characteristics. The supply of flesh phytoplankton-derived labile organic matter into the bottom layer could be expected to promote rapid bacterial decomposition and contribute to the formation of hypoxic water masses in early summer when the ocean was strongly stratified. Atmospheric deposition-derived nutrients in East Asia will have important impacts on not only the oligotrophic outer ocean but also surrounding coastal areas in the warm rainy season.

To identify the origin of various fishes and reconstruct their migration history at the individual level, isotope analysis is a powerful alternative to artificial tagging. We used a novel individual-based methodology to reconstruct individual migratory and/or trophic shifts associated with growth based on isotopic data in the vertebral centrum of adult Japanese flounder Paralichthys olivaceus in Sendai Bay. We measured carbon and nitrogen isotope ratios (δ^13C and δ^15N) in muscle tissues, and conducted a segmental isotope analysis of bulk δ^13C (δ^13Cbulk), bulk δ^15N (δ^15Nbulk), and δ^15N of glutamic acid (δ^15NGlu) and phenylalanine (δ^15NPhe) in vertebral collagen. The δ^15NGlu and δ^15NPhe values for bone collagen revealed an increase in trophic position and a shift to lower trophic baselines (δ^15NBase: indicative of δ^15N values of primary trophic sources) for most individuals. For both δ^13Cbulk and δ^15Nbulk, we detected significant positive correlations between values for muscle and the outermost section of vertebral collagen. A nonlinear time-series analysis of δ^13Cbulk and δ^15Nbulk suggested that a combination of intrinsic (the timing of migration from the nursery to deep offshore areas in juveniles) and extrinsic (habitat and/or food qualities) factors influence the isotopic chronology. A segmental isotope analysis revealed the segregation of individuals among sampling sites at all life stages and changes in trophic positions and δ^15NBase values during growth. Our results suggest that the P. olivaceus population in Sendai Bay has both temporal and spatial structure. The temporal structure may be caused by variation in the timing of migration from the nursery to the deep offshore area in juveniles, and the spatial structure may be explained by individual variation in habitat preferences.

Rationale Carbon stable isotope analysis of bulk samples and fatty acids is an established method for tracing carbon flow pathways and reconstructing trophic interactions, but there is no consensus on which sample drying method should be used for sample preparation. The aim of this study was to determine if freeze-drying and oven-drying treatments used to prepare samples of the benthic macroinvertebrates Stenopsyche marmorata and Epeorus latifolium for bulk and fatty-acid-specific carbon stable isotope analysis yield different isotopic ratio values. Methods Five individuals each from two species were split in half; one half was freeze-dried and the other half was oven-dried. The samples were ground and the δ(13)C values of the bulk samples and eight fatty acids were measured following combustion using an isotope ratio mass spectrometer coupled to an elemental analyzer or gas chromatography system. Results The mean difference in the bulk and fatty acid δ(13)C values between freeze-dried and oven-dried samples was small (≤0.1‰ in both cases), although relatively large variations were observed in individual fatty-acid-specific δ(13)C values (maximum of ≤0.9 ‰). There were no significant differences in either bulk sample or fatty-acid-specific δ(13)C values between freeze-dried or oven-dried samples of the same species. Conclusions Freeze-drying and oven-drying are equally acceptable methods for preparing freshly caught S. marmorata and E. latifolium samples for bulk and fatty-acid-specific carbon stable isotope analyses.

Isotopic compositions of animal tissue are an intrinsic marker commonly used to trace animal origins and migrations; however, few isotopes are effective for this purpose in marine environments, especially on a local scale. The isotope ratio of the lanthanoid element neodymium (Nd) is a promising tracer for coastal animal migrations. Neodymium derives from the same geologic materials as strontium, well known as an isotopic tracer (87Sr/86Sr) for terrestrial and anadromous animals. The advantage of the Nd isotope ratio (143Nd/144Nd, expressed as εNd) is that it varies greatly in the ocean according to the geology of the neighboring continents, whereas oceanic 87Sr/86Sr is highly uniform. This study explored the utility of the Nd isotope ratio as a marine tracer by investigating the variation of εNd preserved in tissues of coastal species, and the causes of that variation, in a region of northeastern Japan where the bedrock geology is highly variable. We measured εNd and 87Sr/86Sr in seawater, river water, and soft tissues of sedentary suspension feeders: the mussels Mytilus galloprovincialis and Mytilus coruscus and the oyster Crassostrea gigas. We also measured concentrations of three lanthanoids (La, Ce, and Pr) in shellfish bodies to determine whether the Nd in shellfish tissue was derived from solution in seawater or from suspended particulates. The εNd values in shellfish tissue varied regionally (−6 to +1), matching the ambient seawater, whereas all 87Sr/86Sr values were homogeneous and typical of seawater (0.7091–0.7092). The seawater εNd values were in turn correlated with those in the adjacent rivers, linking shellfish εNd to the geology of river catchments. The depletion of Ce compared to La and Pr (negative Ce anomaly) suggested that the Nd in shellfish was derived from the dissolved phase in seawater. Our results indicate that the distinct Nd isotope ratio derived from local geology is imprinted, through seawater, on the soft tissues of shellfish. This result underscores the potential of εNd as a tracer of coastal marine animals.

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.

To estimate the effectiveness of preservative/poisons on sediment trap material in freshwater environments, we measured the ratio of particulate organic matter retention [particulate organic carbon (POC) and nitrogen (PON), particulate phosphorus (PP)] using simulated sediment trap material (SSTM). SSTM was collected in summer and fall from depths of 30 and 85 m in the mesotrophic northern basin of Lake Biwa, Japan. All SSTM was incubated in buffered formalin, sodium azide (NaN3) and mercuric chloride (HgCl2) solutions for a period of 56 days. The retention ratio of POC, PON and PP of SSTM was measured throughout the course of the experiments. Results showed that buffered formalin and HgCl2 were very effective in preserving POC. Although NaN3 was less effective at preventing degradation of POC, the degradation process as a function of time was described with the double-exponential decay equation extremely well. Buffered formalin and HgCl2 were also effective in preserving PON, but less effective than for POC. To ensure accurate measurements of PON vertical flux, therefore, one should estimate the decomposition kinetics of PON in sediment trap material. None of the preservatives used in this study was effective for preserving PP, which was rapidly degraded within a week. However, the degradation process was also fully described by the equation, and this will make it possible to accurately estimate PP vertical flux.

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.

The northern East China Sea (ECS) serves as a spawning and nursery ground for many species of fish and squid. To clarify the basis of the food web in the northern ECS, we examined the nitrate (NO3) dynamics along four latitudinal transects based on stable nitrogen and oxygen isotopes of NO 3 (δ15NNO3 and δ18ONO3) and temperature-salinity dynamics in both winter (February 2009) and summer (July 2009 and July 2011). The δ15NNO3 and δ18ONO3, which were distinctly different among the potential NO3 sources, were useful for clarifying NO3 sources and its actual usage by phytoplankton. In winter, Kuroshio Subsurface Water (KSSW) and the Yellow Sea Mixed Water (YSMW) predominantly contributed to NO3 distributed in the shelf water. In the surface water of the Okinawa Trough, NO3 from the KSSW, along with a temperature increase caused by an intrusion of Kuroshio Surface Water (KSW), seemed to stimulate phytoplankton growth. In summer, Changjiang Diluted Water (CDW), Yellow Sea Cold Water Mass (YSCWM), and KSSW affected the distribution and abundance of NO 3 in the northern ECS, depending on precipitation in the Changjiang drainage basin and the development of the YSCWM in the shelf bottom water. Although isotopic fractionation during NO3 uptake by phytoplankton seemed to drastically increase δ15NNO3 and δ18ONO3 in summer, relatively light nitrate with δ15NNO3 lower than expected from this fractionation effect might be explained by contribution of atmospheric nitrogen and/or nitrification to NO3 dynamics in the surface and subsurface layers. If the latter were a dominant process, this would imply a tightly coupled nitrogen cycle in the shelf water of the northern ECS., Biogeosciences, 11(4), pp.1297-1317; 2014

The dynamics of fluorescent dissolved organic matter (FDOM) in the large monomictic freshwater Lake Biwa (surface area 675 km2, maximum depth 104 m) was studied from December 2010 to December 2011. The protein-like FDOM (FDOMT) and dissolved organic carbon (DOC) showed epilimnetic accumulation (FDOMT from 4.42 ± 0.22 quinine sulfate units [QSU] to 6.30 ± 0.04 QSU; DOC from 80.8 ± 2.7 µmol L−1 to 102.7 ± 3.5 µmol L−1) between nutrient-replete winter mixing to nutrient-depleted stratified periods. This accumulation is attributed to the reduced heterotrophic activity following severe P-limitation. The positive correlation between accumulated DOC and FDOMT in the epilimnion and their uniform reduction in the hypolimnion (∼ 9%) suggest FDOMT as a proxy for semi-labile DOM. The humic-like FDOM (FDOMM) generally increased with depth, a pattern similar to nutrients and total carbon dioxide (TCO2), but adverse to dissolved oxygen. The significant positive correlations of FDOMM with apparent oxygen utilization (r = 0.86, p < 0.001), TCO2 (r = 0.91, p < 0.001), nitrate (r = 0.83, p < 0.001), and phosphate (r = 0.76, p < 0.001) in the deeper layers suggest that FDOMM is formed during hypolimnetic mineralization. We estimated that ∼ 8% of the organic carbon degraded in the hypolimnion is transferred into humic substances. The minor contribution of DOC (6.4%) to hypolimnetic mineralization suggests that production of humic substances is mainly fueled by the mineralization of sinking biogenic particles. The production and consumption of FDOM in freshwater lakes may influence the quality and bioavailability of carbon exported from these systems.

Dissolved organic nitrogen (DON) potentially plays a major role in sustaining the high productivity and biological diversity of coral reefs. However, data are scarce regarding sources and sinks of DON. This study, for the first time, determined the 15N isotopic composition of total dissolved nitrogen (δ15NTDN), reflecting the isotopic signature of DON, in the water column over a coral reef. The uniformity in δ15NTDN during high tide (3.2 ± 0.3 ‰) indicated that the DON was mainly derived from offshore waters. In contrast, higher spatial heterogeneity of δ15NTDN (3.1 ± 0.9 ‰) and DON concentrations during low tide indicated the existence of local DON sources patchily distributed over the reef. Low δ15NTDN values located mid-reef were indicative of DON release from organisms that obtained their N via N2 fixation, whereas high δ15NTDN appeared to reflect localized release of DON by organisms exposed to dissolved inorganic nitrogen with elevated 15N, such as from terrestrial and offshore inputs. Collectively, the results highlight the importance of spatial patterns in DON release from reef communities in the N cycling of coral reefs.

Identifying sources of particulate organic matter (POM) is important for clarifying fundamental mechanisms by which lake food webs are sustained. We determined carbon and nitrogen stable isotope ratios of POM in near-shore waters of Lake Biwa, a large, meso-eutrophic lake in Japan, to estimate relative contributions of terrigenous particulate organic carbon (T-POC), plankton-derived POC (P-POC) and epilithon-derived POC (E-POC) to POC in near-shore waters. Samples were collected during different months (November, February, May and July) at 29 sites located near the mouth of tributary rivers with different discharge and catchment land use. The data revealed that POC mainly consisted of P-POC and T-POC, with relative contributions varying widely over season and among locations. E-POC generally contributed little to the near-shore POC. Path analyses revealed that the concentration of riverine POC whose isotopic signatures were similar to those of rice straws increased with a larger %paddy field area in the catchment of tributary rivers, which subsequently enhanced T-POC inputs to near-shore waters through riverine transportation. Furthermore, our results suggested that T-POC contribution was influenced, with a time lag, by wave-driven turbulence and shore topography, which appear to affect sedimentation and resuspension of T-POC.

Stable nitrogen and oxygen isotope ratios of nitrate (δ(15)N-NO(3) and δ(18)O-NO(3)) have recently been used to identify nitrogen sources in water environments. However, there have been no investigations designed to determine nitrate isotopes in non-point sources in urban areas for evaluating the impact of surface deposits on nitrogen in surface runoff. In this study, we collected rainwater, surface runoff and surface deposits (road dust, roof dust and soil) to evaluate the nitrogen sources in surface runoff using nitrate isotopes. There were no large differences in δ(15)N-NO(3) among rainwater (-0.3‰ to 1.5‰), surface runoff (-2.7‰ to 0.4‰), leachates from road dust (-5.8‰ to 6.2‰) and soil (-11.5‰ to 0.6‰). In contrast, the δ(18)O-NO(3) in surface runoff (28.5-47.9‰) was lower than that in rainwater (62.7-78.6‰), and higher than that in leachates from road dust (6.1-27.6‰) and soil (-1.1‰ to 6.6‰). δ(18)O-NO(3) is a useful indicator for evaluating the NO(3)-N sources in surface runoff. Using this indicator, NO(3)-N from road dust was estimated to account for more than half of the NO(3)-N in surface runoff. This is consistent with a result based on a comparison of their loads per unit surface between rainwater and surface runoff, which also showed that most of the nitrogen in surface runoff was derived from surface deposits.

The Tuul River, which provides water for the daily needs of many residents of Ulaanbaatar, Mongolia, has been increasingly polluted by wastewater from the city's sewage treatment plant. Information on water movement and the transformation of water-borne materials is required to alleviate the deterioration of water quality. We conducted a synoptic survey of general water movement, water quality including inorganic nitrogen concentrations, and isotopic composition of nitrogen (δ 15 N-NO 3 − , δ 18 O-NO 3 − , and δ 15 N-NH 4 + ) and water (δ 18 O-H 2 O) in a wetland area that receives wastewater before it enters the Tuul River. We sampled surface water, groundwater, and spring water along the two major water routes in the wetland that flow from the drain of the sewage treatment plant to the Tuul River: a continuous tributary and a discontinuous tributary. The continuous tributary had high ammonium (NH 4 + ) concentrations and nearly stable δ 15 N-NH 4 + , δ 15 N-NO 3 − , and δ 18 O-NO 3 − concentrations throughout its length, indicating that nitrogen transformation (i.e., nitrification and denitrification) during transit was small. In contrast, NH 4 + concentrations decreased along the discontinuous tributary and nitrate (NO 3 − ) concentrations were low at many points. Values of δ 15 N-NH 4 + , δ 15 N-NO 3 − , and δ 18 O-NO 3 − increased with flow along the discontinuous route. Our results indicate that nitrification and denitrification contribute to nitrogen removal in the wetland area along the discontinuous tributary with slow water transport. Differences in hydrological pathways and the velocity of wastewater transport through the wetland area greatly affect the extent of nitrogen removal.

Summary Deterioration of the quality of groundwater under urban areas has become a major environmental concern worldwide. This paper presents a pilot case study (in the Taipei urban area) using multiple stable isotopes ( δ D H 2 O and δ 18 O H 2 O , δ 15 N NO 3 and δ 18 O NO 3 , δ 34 S SO 4 and δ 18 O SO 4 , and 87Sr/86Sr) with the aim of understanding the subsurface nature and environmental status of such areas. A previous study has already characterized Taipei shallow groundwaters as sulfate-rich in comparison to other Southeast Asian metropolitan cities. Our chemical measurements clarified that the arsenic is the important environmental concerns in the aquifers. Dual isotope approaches for nitrate and sulfate suggested these ions were sourced mainly from municipal sewage leaking from sewer pipes in urbanized area, while chemical fertilizers applied in the local agricultural fields partly contribute in the suburb area. Significant decreases in nitrate and sulfate concentrations due to denitrification and sulfate reduction were observed in some groundwaters, which induced increases in isotope ratios up to 24.9‰ for δ 15 N NO 3 and 27.2‰ for δ 34 S SO 4 . Nitrate groundwater pollution in the Taipei Basin is not a subject of serious concern ( NO 3 - for all analyzed samples) at the present time due to attenuation by denitrification. However, arsenic is preferentially released in groundwater (>100 μg/L) in anoxic environment under which sulfate reduction occurs. The Sr isotope data suggested that the arsenic is derived from alluvial deposits of Oligocene–Miocene sedimentary rock origin. This study disclosed the complex causes of environmental changes in Taipei urban shallow groundwater. It is also important as an excellent case study demonstrating the efficient use of multiple isotope diagnosis in the urban groundwater environmental field.

Carbon and nitrogen stable isotope ratios (δ13C and δ15N) of macroinvertebrates inhabiting littoral zones of lakes can serve as useful indicators of material loading from the watershed. We collected snails (Semisulcospira spp.) and bivalves (Unio douglasiae biwae Kobelt) from 29 littoral sites in Lake Biwa near the mouths of river tributaries with various human population density (HPD) and land-use patterns. The δ13C and δ15N signatures were determined for three potential food sources: particulate organic matter in the pelagic zone (PPOM), riverine particulate organic matter from tributaries (RPOM) and epilithic organic matter in the littoral zone (EOM). The stable isotope mixing model revealed that snails relied mainly on EOM, and bivalves on PPOM and RPOM. Multiple regression analysis showed that intersite variation in δ15N for snails was best explained by HPD, while variation in δ15N of EOM and nitrate was explained to a lesser extent by HPD. Comparison with isotope signatures of their food sources and riverine nutrients revealed that snails assimilated anthropogenic nitrogen from wastewater in the watershed. Our results also showed that the δ13C value of bivalves was marginally related to the fraction of paddy fields in the watersheds. In conclusion, the isotope signatures of macroinvertebrates inhabiting the littoral zone can be useful indicators of anthropogenic impacts from the watershed.

Detailed cross-sectional temperature and oxygen distributions during winter were examined over 5 years (2003–2007) in large, warm, monomictic Lake Biwa. The distribution patterns showed year-to-year variation in the degree of spatial heterogeneity and indicated that both convective mixing and gravity currents contributed to the delivery of oxygen to the profundal zone. We encountered a warm winter in 2007, when deep-water oxygenation was delayed by more than a month relative to normal years. Data obtained during the period of oxygenation in 2007 suggest that the timing of complete oxygenation of the bottom water (or lower layers of the hypolimnion) was controlled by physical processes responsible for destruction of the weak thermal stratification near the bottom. This study revealed that Lake Biwa is sensitive to year-to-year variations in winter meteorological conditions. Specifically, the bottom environments are highly susceptible to a warm winter, especially when it follows a cold winter.

Semi-labile dissolved organic carbon (DOC) plays an important role in the transport and hypolimnetic remineralization of carbon in large freshwater lakes. However, sources of semi-labile DOC in lakes remain unclear. This study used a carbon stable isotope approach to examine relative contributions of autochthonous and allochthonous sources to semi-labile DOC. Vertical and seasonal variations in the concentration and carbon stable isotope ratio (δ13C) of DOC were determined in large (surface area 674 km2; maximum depth 104 m), monomictic Lake Biwa. A sharp vertical gradient of δ13C of DOC (δ13C-DOC) during the stratification period [mean ± standard error (SE) −25.5 ± 0.1 and −26.0 ± 0.0‰ in the epi- and hypolimnion, respectively] indicated the accumulation of 13C-rich DOC in the epilimnion. Vertical mixing explained the intermediate values of δ13C-DOC (−25.7 ± 0.0‰) measured throughout the water column during the overturn period. Both DOC concentration and δ13C-DOC decreased in the hypolimnion during stratification, indicating selective remineralization of 13C-rich DOC. Using a two-component mixing model, we estimated the δ13C value of semi-labile DOC to be −22.2 ± 0.3‰, which was close to the δ13C of particulate organic carbon collected in the epilimnion during productive seasons (−22.7 ± 0.7‰) but much higher than the δ13C-DOC in river waters (−26.5 ± 0.1‰). Semi-labile DOC appeared to be mainly autochthonous in origin, produced by planktonic communities during productive seasons. The spatiotemporal uncoupling between production and remineralization of semi-labile DOC implies that hypolimnetic oxygen consumption may be affected by pelagic primary production during productive seasons of the preceding year.

Epilithic bacterial communities play a major role in biogeochemical cycles of rivers; however, distributional patterns and controls of epilithic communities remain unclear. The objective of the present study was to examine possible environmental factors that affect longitudinal distri- butional patterns of epilithic bacterial communities in 2 rivers (Yasu and Ado Rivers) draining the Lake Biwa basin, Japan. Phylogenetic analyses were conducted using denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes and fluorescence in situ hybridization (FISH) with oligonucleotide probes targeted to small subunit rRNA. Non-metric multidimensional scaling and canonical correspondence analysis of the DGGE profile indicated that a clear shift in community composition occurred at the middle reach of the Yasu River. This shift was most closely related to variation in the nitrogen stable isotope ratio (δ 15 N) of epilithic materials (BIOENV analysis, ρw = 0.512), which is an indicator of the extent of anthropogenic nitrogen loading. The concentration of dissolved organic carbon (DOC) also affected community composition (ρw = 0.510 for the combina- tion of δ 15 N and DOC). The longitudinal pattern of the DGGE profile was less evident in the Ado River. In both rivers, FISH results indicated that Alphaproteobacteria and Betaproteobacteria gener- ally accounted for a large fraction (37 to 71%) of total bacterial abundance, indicating that species and clones affiliated with a limited number of the major phylogenetic groups occupy diverse epilithic habitats. Our results are consistent with the hypothesis that nutrients and organic matter of anthropo- genic origin are major determinants of epilithic bacterial community structure in rivers.

Deterioration of aquatic ecosystems resulting from enhanced anthropogenic N loading has become an issue of increasing concern worldwide, and methods are needed to trace sources of N in rivers. Because nitrate from sewage is enriched in 15N relative to nitrate from natural soils, delta(15)N values of stream nitrate (delta(15)Nnitrate) should be an appropriate index of anthropogenic N loading to rivers, as should the delta(15)N values of riparian plants (delta(15)Nplant) because they are consumers of nitrate. We determined the delta(15)N values of stream nitrate and six species of riparian macrophytes in 31 rivers in the Lake Biwa Basin in Japan. We then tested the correlation between these values and various land-use parameters, including the percentage of land used for residential and agricultural purposes as well as for natural areas. These delta(15)N values were significantly positively correlated with land use (%) that had a high N load (i.e., residential or agricultural use) and significantly negatively correlated with forest (%). These findings indicate that delta(15)N values of stream nitrate and riparian plants might be good indicators of anthropogenic inputs of nitrogen.

We examined how nitrogen-stable isotopic signatures of food web components (basal resources, primary and lower consumers, and omnivores) in rivers change with increasing levels of human population density (HPD) in their watersheds. Samples were collected from 22 rivers flowing in the Lake Biwa basin, Japan. Among three potential resources at the base of food webs (epilithon, benthic and suspended particulate organic matter), the mean isotopic values (δ15N) of the epilithon (4.5–7.8%) were consistently higher than those of other items (1.9–4.2%) and displayed the most pronounced elevation (by 3.3%) with increasing HPD. The mean δ15N values of the individual taxa of lower consumers (bivalve, snail and caddisfly) tended to increase with increasing HPD, although the pattern and the extent of the elevation were highly variable among the taxa. These results suggest a taxon-specific feature in the N source (or sources) of lower consumers. Our data suggested that human activities (e.g. nutrient loading) potentially induce changes in the N baselines of river food webs. The major N source of bivalves appeared to be shifted from suspended particulate organic matter to other items with increasing HPD. Trophic levels of goby fish (Rhinogobius sp. OR) and shrimp (Palaemon paucidens), being estimated to be at 2.4–3.8 and 2.1–3.4, respectively, did not differ significantly among rivers with different HPD levels.

Seasonal production of bacterioplankton in a water column of Lake Biwa was examined based on cell-specific growth rates at above (surface layer) and below the thermocline (deep layer). The growth rates were estimated by incubating bacterioplankton in situ with a dilution technique. The rates ranged from 0.05 per day in winter to 0.89 per day in summer, generally with much higher rates in the surface layer than in the deep layer. In an entire water column, bacterial production (in terms of carbon [C]) ranged from 0.217 to 0.811 gC/m2 per day with a mean of 0.451 gC/m2 per day, which, on average corresponded to 43% of primary production. However, no significant correlation was detected between bacterial production and primary production rates. Although the bacterial production rate correlated positively to water temperature, surface and deep layer rates were comparative for some dates because the deep layer shared a large fraction of the water column. These results suggest that, although specific bacterial activity was low in the deep layer due to the low temperature, bacteria in that layer play substantial roles in consumption of organic matter and material flows in Lake Biwa.

To examine the role of Daphnia in the decomposition and elimination of organic carbon at the surface layer, organic matter from Lake Biwa was incubated with and without Daphnia at 20 � C in the dark for 15 days in May, July and October. In all of the experiments, total organic carbon decreased during the incubation, regardless of treatment; however, the net effect of Daphnia on this decrease differed among experiments. Daphnia stimulated the decrease in particulate organic carbon in two out of three experiments. Bacterial abundance was higher in treatments with Daphnia than in treatments without Daphnia in all experiments. Higher bacterial abundance in treatments with Daphnia could not be explained fully by zooplankton grazing effects on phagotrophic protozoans, rather Daphnia seemed to stimulate bacterial growth by supplying substrates for that growth. The results of the present study suggest that Daphnia reduces sinking flux, not only by direct grazing, but also by the channelling of particulate organic carbon into dissolved forms that are then available for bacterial growth.

(2002). Carbon and phosphorus budgets in the pelagic area of Lake Biwa, the largest lake in Japan. SIL Proceedings, 1922-2010: Vol. 28, No. 3, pp. 1409-1414.

We performed bag experiments in a Canadian Shield lake with generally high seston (suspended food particles mainly composed of algae) carbon (C):phosphorus (P) ratios, and investigated the responses of individual and population growth of herbivorous Daphnia dentifera on their abundance with (+ P) and without (-P) a phosphorus enrichment to lake water. In both treatments, increased abundance of D. dentifera reduced seston C concentration and was accompanied by decreases in population and individual growth rates. However, P-enrichment increased seston P concentration and then reduced seston C:P ratio from 400-700 to ca 100 (by atoms). As a result, both individual and population growth rates were significantly higher in the + P treatment at all animal abundances even though seston C concentrations were similar between the treatments. The magnitude of the growth enhancement by the P-enrichment was independent of animal abundance. Stepwise regression analyses revealed that 71 and 90% of the variance in the population and individual growth rates, respectively, were explained by seston C and P concentrations, and that the contribution of the seston P concentration was roughly the same as that of seston C. Such joint effects of seston C and P indicate that food quality (P content) as well as food quantity (C concentration) can influence Daphnia not only at the level of individual growth but also at the level of population dynamics in P-limited lakes. Our results thus strongly corroborate the hypothesis that the population development of a key herbivore Daphnia in P-limited Canadian Shield lakes is inhibited by the direct effects of P-limited food on individual growth, which weaken the strength of trophic cascading interactions starting from piscivorous fish through planktivorous fish and zooplankton to algae.

Nitrogen (N) saturation of upland forests has been assumed to be a substantial N source downstream. However, removal processes of N, including assimilation and denitrification in the downstream area, have not been clarified. To evaluate the N removal processes, nitrate (NO3−) and organic N concentrations, as well as nitrogen isotope ratio (δ15N) and oxygen isotope ratio (δ18O) of NO3− were measured along three rivers of Tatara River Basin, Japan where upland forests have already been N-saturated. Geographic information system (GIS) based topographical analysis was also conducted to evaluate the land use as urban area in relation to topography. In two of the three rivers, NO3− concentrations did not increase from upstream to downstream, despite the potential non-point N sources of urban areas. In another river, NO3− concentrations rather decreased. The values of δ15N and δ18O of NO3− and organic N concentrations suggested the presence of denitrification and assimilation over N pollutants in the river whose watersheds have a lower percentage of urban area. The lower percentage of urban area could be explained by the lower topographic index. This study concluded that the NO3− leaching from upland N-saturated forests was substantially assimilated or denitrified in the downstream area.

To clarify the roles of zooplankton in the sedimentation of seston from the epilimnion, the sinking flux of particulate carbon was measured along with primary production rate and zooplankton biomass from July 1996 to October 1997 at a pelagic site in the north basin of Lake Biwa. During the study period, the flux varied seasonally from 66 to 510 mg C m−2 day−1 and was low in summer when zooplankton, composed mainly of Eodiaptomus japonicus and Daphnia galeata, were abundant. Simple correlation analysis revealed that the sinking flux correlated neither with the primary production rate nor with the amount of sestonic carbon above the sediment trap. However, the particle elimination rate, estimated as the difference between the primary production rate and the sinking flux, correlated positively with the zooplankton biomass. These results suggest that zooplankton play a substantial role in decreasing the sinking flux in Lake Biwa.

Simple correlation and multiple regression analyses were performed to examine the relationship between primary productivity and environmental factors in the north basin of Lake Biwa. The primary production rates used in the analyses were estimated monthly or bimonthly during the growing season (April‐November) in 1992, 1996 and 1997 with the 13 C method. Elemental (C, N and P) contents of seston were used to assess nutrient conditions. Analyses revealed that 86% of variance in depth-integrated primary production rates (areal PP) can be explained by changes in light intensity, and sestonic C, N and P concentrations. Water temperature had no effect on areal PP. To assess relative effects of light and nutrients on PP, the P:B ratio was estimated by normalizing PP with sestonic C. The areal P:B ratio correlated most significantly with the sestonic N:P ratio, followed by light intensity. When regression analyses were made at each depth, however, the P:B ratio correlated significantly only with the sestonic N:P ratio at 0 and 1 m depths, while light intensity was also incorporated into the regressions at deeper than 2.5 m. In these regressions, the P:B ratio was negatively correlated with sestonic N:P ratio but positively with light intensity. The results suggest that the primary production rate in this lake was mainly limited by P relative to N supply rates, but was not free from light limitation in a large part of the epilimnion. In Lake Biwa, the vertical water mixing regime as well as the nutrient supply seem to be important in determining the growth and composition of primary producers, since the surface mixing layer extends into 10‐15 m depths during most of the growing season.

[1] To clarify the pathways and origins of dissolved nitrate (NO3−) in rivers flowing into Lake Biwa, Japan, three types of coordinated surveys of concentrations and isotope compositions of NO3− were conducted: (1) synoptic river sampling of 32 representative inflow rivers, (2) two rivers in catchments with different land uses, and (3) headwater catchment samplings. The data set used in this paper is the first comprehensive collection of isotopic composition of NO3− in rivers of a large-scale basin in Asia. While the NO3− spatial concentration pattern showed significant seasonal variation, the spatial pattern of the δ15N of dissolved NO3− was more consistent throughout the year. The was significantly positively correlated with the population density of each catchment. A mass balance model assuming the and the flow rate of sewage effluent was developed. The model simulated the relationship between the population density and the reasonably well, suggesting that the dominant source contributing to the increase in was the sewage effluent. The spatiotemporal distribution of suggested the possibility of the addition of atmospherically derived NO3− through precipitation and was indicated by the hydrological pathways for NO3− exports in different land uses. In forest-dominated catchments with natural drainage systems, a slightly elevated δ18ONO3 signal remained in the stream water even during base flow conditions. This study demonstrated that multiscale, multi-isotopic investigation is a promising strategy for describing the spatial distribution of NO3− sources synoptically and is useful for evaluating the influences of land use change.

[1] To elucidate the internal nitrogen cycle and nitrogen input and output, we measured δ15N-NO3−, δ18O-NO3−, and NO3− concentrations in rainfall, soil water from an unsaturated soil horizon, groundwater, and stream water at a small forested headwater catchment. In soil water, the mean NO3− concentration did not decrease concomitantly with soil depth, despite a drastic decrease of δ18O-NO3− with soil depth, indicating immediate consumption of atmospheric NO3− and production of nitrified NO3−. The spatiotemporal distribution of δ15N-NO3− and δ18O-NO3− in soil water indicated that plant uptake contributed mainly to NO3− consumption and that denitrification was negligible. In contrast, denitrification played a critical role in groundwater in decreasing the NO3− concentration. Fluctuations of δ15N-NO3−, δ18O-NO3−, and NO3− concentration in groundwater, however, differed from expectations for occurrence resulting from denitrification only. A plausible explanation for the lack of a concurrent increase in δ15N and δ18O of NO3− is that nitrification and denitrification occur concurrently in groundwater and that denitrification is more important than nitrification. The amount of denitrification in the groundwater body was controlled primarily by groundwater residence time. The combination of hydrological flow path from groundwater to stream water and denitrification in the groundwater critically control the amount of leaching of NO3− and temporal distribution of NO3− concentration in stream water. The possibility exists that denitrification in groundwater strongly influences nitrogen leaching in forests because denitrification occurred even in this catchment, which consists of weathered granitic bedrock and predominantly typic udipsamment, with low organic matter content in the soil.

The status of nitrate (NO 3 − ), nitrite (NO 2 − ) and ammonium (NH 4 + ) contamination in the water systems, and the mechanisms controlling their sources, pathways, and distributions were investigated for the Southeast Asian cities of Metro Manila, Bangkok, and Jakarta. GIS-based monitoring and dual isotope approach (nitrate δ 15 N and δ 18 O) suggested that human waste via severe sewer leakage was the major source of nutrient contaminants in Metro Manila and Jakarta urban areas. Furthermore, the characteristics of the nutrient contamination differed depending on the agricultural land use pattern in the suburban areas: high nitrate contamination was observed in Jakarta (dry fields), and relatively lower nutrients consisting mainly of ammonium were detected in Bangkok (paddy fields). The exponential increase in NO 3 − – δ 15 N along with the NO 3 − reduction and clear δ 18 O/ δ 15 N slopes of NO 3 − (∼ 0.5) indicated the occurrence of denitrification. An anoxic subsurface system associated with the natural geological setting ( e.g. , the old tidal plain at Bangkok) and artificial pavement coverage served to buffer NO 3 − contamination via active denitrification and reduced nitrification. Our results showed that NO 3 − and NH 4 + contamination of the aquifers in Metro Manila, Bangkok, and Jakarta was not excessive, suggesting low risk of drinking groundwater to human health, at present. However, the increased nitrogen load and increased per capita gross domestic product (GDP) in these developing cities may increase this contamination in the very near future. Continuous monitoring and management of the groundwater system is needed to minimize groundwater pollution in these areas, and this information should be shared among adjacent countries with similar geographic and cultural settings.

The status of nitrate (NO(3)(-)), nitrite (NO(2)(-)) and ammonium (NH(4)(+)) contamination in the water systems, and the mechanisms controlling their sources, pathways, and distributions were investigated for the Southeast Asian cities of Metro Manila, Bangkok, and Jakarta. GIS-based monitoring and dual isotope approach (nitrate delta(15)N and delta(18)O) suggested that human waste via severe sewer leakage was the major source of nutrient contaminants in Metro Manila and Jakarta urban areas. Furthermore, the characteristics of the nutrient contamination differed depending on the agricultural land use pattern in the suburban areas: high nitrate contamination was observed in Jakarta (dry fields), and relatively lower nutrients consisting mainly of ammonium were detected in Bangkok (paddy fields). The exponential increase in NO(3)(-)-delta(15)N along with the NO(3)(-) reduction and clear delta(18)O/delta(15)N slopes of NO(3)(-) ( approximately 0.5) indicated the occurrence of denitrification. An anoxic subsurface system associated with the natural geological setting (e.g., the old tidal plain at Bangkok) and artificial pavement coverage served to buffer NO(3)(-) contamination via active denitrification and reduced nitrification. Our results showed that NO(3)(-) and NH(4)(+) contamination of the aquifers in Metro Manila, Bangkok, and Jakarta was not excessive, suggesting low risk of drinking groundwater to human health, at present. However, the increased nitrogen load and increased per capita gross domestic product (GDP) in these developing cities may increase this contamination in the very near future. Continuous monitoring and management of the groundwater system is needed to minimize groundwater pollution in these areas, and this information should be shared among adjacent countries with similar geographic and cultural settings.

a Research Institute for Humanity and Nature, Japan b Department of Earth Science and Technology, Akita University, Japan c Faculty of Integrated Arts and Sciences, Hiroshima University, Japan d Marine Science Institute, University of the Philippines, Philippines e Department of Ground Water Resources, Ministry of Natural Resources and Environment, Thailand f Division of Hydrology, Indonesia Institute of Sciences, Indonesia g CREST, Japan Science and Technology Agency, Saitama, Japan h Center for Ecological Research, Kyoto University, Japan

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 (