Catchment land-use, macroinvertebrates and detritus processing in headwater streams: taxonomic richness versus function

1. We used a litter bag technique to assess the effect of catchment land-use (forest, wetland, agriculture, urban) on the processing of red maple ( Acer rubrum L.) litter in 17 streams in Maine, U.S.A. Litter processing by fungi was predicted to increase with nutrient concentrations along a gradient of land use, from relatively unmodified to highly modified. Litter processing by litter-shredding macroinvertebrates was predicted to decline along this gradient because of a decline in their taxonomic richness and biomass.
2. Land use was associated with the anticipated gradient in nutrient and macroinvertebrate attributes, and a significant relationship was found between land use and nitrate concentration. There was, however, no significant relationship between land use and soluble reactive phosphorus (SRP) concentration. Similarly, shredder taxonomic richness was significantly related to land use type, whereas shredder biomass showed no significant relationship to land use.
3. Attributes of the shredder assemblage structure and nutrient concentrations were both strong determinants of litter processing. Increasing biomass and taxonomic richness of shredders was significantly related to increasing rates of litter mass loss. Increasing concentrations of nitrate and SRP were significantly related to increasing rates of litter softening below threshold concentrations (approximately 0.20 mg NO₃-N L^–1 and 5 μg SRP L^–1).
4. The potentially additive effects of nitrate and SRP concentrations or shredder richness and biomass on litter processing rates were confounded by the lack of significant correlation between these pairs of variables. Consequently, rates of litter processing (as rates of softening or mass loss) did not vary systematically among different land use regimes.     

The impact of climate change on the structure of Pleistocene food webs across the mammoth steppe

Species interactions form food webs, impacting community structure and, potentially, ecological dynamics. It is likely that global climatic perturbations that occur over long periods of time have a significant influence on species interaction patterns. Here, we integrate stable isotope analysis and network theory to reconstruct patterns of trophic interactions for six independent mammalian communities that inhabited mammoth steppe environments spanning western Europe to eastern Alaska (Beringia) during the Late Pleistocene. We use a Bayesian mixing model to quantify the contribution of prey to the diets of local predators, and assess how the structure of trophic interactions changed across space and the Last Glacial Maximum (LGM), a global climatic event that severely impacted mammoth steppe communities. We find that large felids had diets that were more constrained than those of co-occurring predators, and largely influenced by an increase in Rangifer abundance after the LGM. Moreover, the structural organization of Beringian and European communities strongly differed: compared with Europe, species interactions in Beringian communities before—and possibly after—the LGM were highly modular. We suggest that this difference in modularity may have been driven by the geographical insularity of Beringian communities.     

Leaf breakdown in a regulated desert river: Colorado River, Arizona, U.S.A.

We compared processing rates (k _d) for leaves of the native willow (Salix exigua Nutt.) and cottonwood (Populus fremontii Wats.) to those of the non-native salt cedar (Tamarix chinensis Lour.) in the regulated Colorado River, U.S.A. Leaf packs of each species were incubated at Lees Ferry, approximately 26 km below Glen Canyon Dam, Arizona. Leaf packs were processed at 2, 21, 46, 84 and 142-d intervals. Water temperatures remained relatively constant (10 °C, SE ± 1 °C) during the study. There were significant differences in processing rates between species, with P. fremontii showing the fastest breakdown. After 142 d, only 20% of the P. fremontii leaf mass remained, whereas 30% and 52% of leaf masses remained for T. chinensis and S. exigua, respectively. The k _d value for P. fremontii was 0.0062 compared to 0.0049 and 0.0038 for T. chinensis and S. exigua, respectively. Invertebrate colonization was not significantly different between native and non-native plant species with oligochaetes the most abundant animal colonizing the leaf packs. Dual stable isotope analysis showed that leaf material was not the primary food for invertebrates associated with leaf packs. Processing rates for all leaf types were slow in the regulated Colorado River compared to rates reported in many other systems. This is likely due to the lack of caddisfly and stonefly shredders and perhaps slow metabolic rates by microbes.     

Food web structure and function in two arctic streams with contrasting disturbance regimes

1. We studied the effect of substratum movement on the communities of adjacent mountain and spring tributaries of the Ivishak River in arctic Alaska (69°1′N, 147°43′W). We expected the mountain stream to have significant bed movement during summer because of storm flows and the spring stream to have negligible bed movement because of constant discharge. 2. We predicted that the mountain stream would be inhabited only by taxa able to cope with frequent bed movement. Therefore, we anticipated that the mountain stream would have lower macroinvertebrate species richness and biomass and a food web with fewer trophic levels and lower connectance than the spring stream. 3. Substrata marked in situ indicated that 57–66% of the bed moved during summer in the mountain stream and 4–20% moved in the spring stream. 4. Macroinvertebrate taxon richness was greater in the spring (25 taxa) than in the mountain stream (20 taxa). Mean macroinvertebrate biomass was also greater in the spring (4617 mg dry mass m^?2) than in the mountain stream (635 mg dry mass m^?2). Predators contributed 25% to this biomass in the spring stream, but only 7% in the mountain stream. 5. Bryophyte biomass was >1000 times greater in the spring stream (88.4 g ash‐free dry mass m^?2) than the mountain stream (0.08 g ash‐free dry mass m^?2). We attributed this to differences in substratum stability between streams. The difference in extent of bryophyte cover between streams probably explains the high macroinvertebrate biomass in the spring stream. 6. Mean food‐web connectance was similar between streams, ranging from 0.18 in the spring stream to 0.20 in the mountain stream. Mean food chain length was 3.04 in the spring stream and 1.83 in the mountain stream. Dolly Varden char (Salvelinus malma) was the top predator in the mountain stream and the American dipper (Cinclus mexicanus) was the top predator in the spring stream. The difference in mean food chain length between streams was due largely to the presence of C. mexicanus at the spring stream. 7. Structural differences between the food webs of the spring and mountain streams were relatively minor. The difference in the proportion of macroinvertebrate biomass contributing to different trophic levels was major, however, indicating significant differences in the volume of material and energy flow between food‐web nodes (i.e. food web function).     

Flow-substrate interactions create and mediate leaf litter resource patches in streams

1. The roles that streambed geometry, channel morphology, and water velocity play in the retention and subsequent breakdown of leaf litter in small streams were examined by conducting a series of field and laboratory experiments. 2. In the first experiment, conditioned red alder (Alnus rubra Bongard) leaves were released individually in three riffles and three pools in a second‐order stream. The transport distance of each leaf was measured. Several channel and streambed variables were measured at each leaf settlement location and compared with a similar number of measurements taken at regular intervals along streambed transects (‘reference locations’). Channel features (such as water depth) and substrate variables (including stone height, stone height‐to‐width ratio, and relative protrusion) were the most important factors in leaf retention. 3. In the second experiment, the role of settlement location and reach type in determining the rate of leaf litter breakdown was examined by placing individual conditioned red alder leaves in exposed and sheltered locations (on the upper and lower edges of the upstream face of streambed stones, respectively) in riffle and pool habitats. After 10 days, percent mass remaining of each leaf was measured. Generally, leaves broke down faster in pools than in riffles. However, the role of exposure in breakdown rate differed between reach types (exposed pool > sheltered pool > sheltered riffle > exposed riffle). 4. In the third experiment, the importance of substrate geometry on leaf litter retention was examined by individually releasing artificial leaves upstream of a series of substrate models of varying shape. Substrates with high‐angle upstream faces (were vertical or close to vertical), and that had high aspect ratios (were tall relative to their width), retained leaves more effectively. 5. These results show that streambed morphology is an important factor in leaf litter retention and breakdown. Interactions between substrate and flow characteristics lead to the creation of detrital resource patchiness, and may partition leaf litter inputs between riffles and pools in streams at baseflow conditions.     

Relationships between the isotopic composition of dissolved organic carbon and its bioavailability in contrasting Ozark streams

Bioavailability of dissolved organic carbon (DOC) can significantly influence nutrient cycling in small streams yet the factors influencing the bioavailability of DOC remains poorly understood. The isotopic composition of DOC and factors controlling its utilization were studied in two contrasting headwater streams to elucidate the relationship between DOC source and its bioavailability. Water samples were collected monthly from Moore Creek (MC), located in a watershed dominated by fertilized pasture, and from Huey Hollow (HH), located in a deciduous forest watershed. Dissolved organic carbon concentrations ranged from 222 to 1130 μm C and 35 to 289 μm C in MC and HH, respectively. The isotopic composition of DOC (δ¹³C_DOC) was more seasonally variable in HH and ranged from ?33.6 to ?28.0?, as compared with MC where it ranged from ?27.2 to ?24.5 ?. The δ¹³C_DOC in Huey Hollow suggests leaf debris was an important source of DOC throughout most of the year while algae was important in winter and early spring. In MC, the δ¹³C_DOC indicated DOC was largely derived from soil organic matter while algal inputs were small relative to the large pool of refractory DOC. Stream water community respiration (SWCR) rates suggest the existence of a larger pool of refractory DOC in MC relative to HH. The ratio of SWCR (μM C h^?1) to DOC concentration (mM C) was generally higher in HH (1.2–13.2) as compared with MC (0.2–4.2), suggesting that relative bioavailability of DOC was often greater in HH. Nutrient addition experiments indicate that bioavailable C limited SWCR in spring and fall in MC, while bioavailable C was never limiting in HH. The results suggest that elevated nutrient and DOC concentrations supported higher levels of microbial activity that resulted in a large pool of refractory DOC in MC. The lower inorganic nutrient and DOC concentrations reduced microbial activity in HH and likely limited the production of refractory DOC. Results of this study suggest that both organic matter source and nutrient concentration are important determinants of DOC bioavailability in small streams. %     

On the vapour trail of an atmospheric imprint in insects

Terrestrial arthropods, at constant risk from desiccation, are highly sensitive to atmospheric temperature and humidity. A physiological marker of these abiotic conditions could highlight phenotypic adaptations, indicate niche partitioning, and predict responses to climate change for a group representing three-quarters of the Earth's animal species. We show that the (18)O composition of insect haemolymph is such a measure, providing a dynamic and quantitatively predictable signal for respiratory gas exchange and inputs from atmospheric humidity. Using American cockroaches (Periplaneta americana) under defined experimental conditions, we show that insects respiring at low humidity demonstrate the expected enrichment in the (18)O composition of haemolymph because of evaporation. At high humidity, however, diffusional influx of atmospheric water vapour into the animal forces haemolymph to become depleted in (18)O. Additionally, using cockroaches sampled from natural habitats, we show that the haemolymph (18)O signature is transferred to the organic material of the insect's exoskeleton. Insect cuticle, therefore, exhibits the mean atmospheric conditions surrounding the animals prior to moulting. This discovery will help to define the climatic tolerances of species and their habitat preferences, and offers a means of quantifying the balance between niche partitioning and 'neutral' processes in shaping complex tropical forest communities.     

Can ecosystem functional recovery be traced to decomposition and nitrogen dynamics in estuaries of the Lower Laguna Madre,Texas?

The biggest incentive to attempt the restoration and protection of estuaries is their widely acknowledged ecological and economic importance. Assessing estuary health and recovery can most accurately come from examining ecosystem processes. The purpose of this study was to explore the potential of mass loss and nitrogen (N) dynamics during leaf litter decomposition, to detect signs of functional recovery in two estuarine systems in south Texas. Submerged litterbags with black mangrove (Avicennia germinans) leaves were retrieved at various dates over 320 days. Decomposition was about 50% slower in one of the recovering systems compared to a reference site. Nitrogen immobilization and release from decaying leaf litter also discriminated among sites. Nitrogen immobilization potentials ranged from 4.15 to 6.89 mgN/g leaf litter, with the reference site exhibiting the highest value and thus the highest potential to conserve N during litter decomposition. The reference site also had a N immobilization time twice as long as the recovering sites, and a slower net release after the immobilization, appearing again as the most conservative in this part of the N cycling, possibly pointing to a less disturbed, or more stable ecosystem. Overall, the N dynamics during decomposition of mangrove leaf litter were similar in both recovering sites, whereas the reference site had a more conservative nutrient dynamics with more N being retained for longer in decomposing litter, coupled with a slower net release. Metrics derived from N dynamics may provide a finer resolution assessment of functional recovery, than only decomposition metrics.     

Overstory Community Composition and Elevated Atmospheric CO2 and O3 Modify Understory Biomass Production and Nitrogen Acquisition

Elevated atmospheric CO₂ and O₃ have the potential to affect the primary productivity of the forest overstory, but little attention has been given to potential responses of understory vegetation. Our objective was to document the effects of elevated atmospheric CO₂ and O₃ on understory species composition and biomass and to quantify nitrogen (N) acquisition by the understory vegetation. The research took place at the aspen free-air CO₂ and O₃ enrichment (FACE) experiment, which has four treatments (control, elevated CO₂, elevated O₃, and elevated CO₂+O₃) and three tree communities: aspen, aspen/birch, and aspen/maple. In June 2003, each FACE ring was uniformly labeled with 15N applied as NH₄Cl. Understory biomass was harvested in June of 2004 for productivity, N, and 15N measurements, and photosynthetically active radiation (PAR) was measured below the canopy. The understory was divided into five species groups, which dominate in this young aggrading forest: Taraxacum officinale (dandelion), Solidago sp. (goldenrod), Trifolium repens and T. pretense (clover), various species from the Poaceae family (grass), and composited minor components (CMC). Understory species composition, total and individual species biomass, N content, and 15N recovery showed overstory community effects, but the direct effects of treatments was masked by the high variability of these data. Total understory biomass increased with increasing light, and thus was greatest under the open canopy of the aspen/maple community, as well as the more open canopy of the elevated O₃ treatments. Species were different from one another in terms of 15N recovery, with virtually no 15N recovered in clover and the greatest amount recovered in dandelion. Thus, understory species composition and biomass appear to be driven by the structure of the overstory community, which is determined by the tree species present and their response to the treatments. However, N acquisition by the understory does not appear to be affected by either the overstory community or the treatments at this point.     

Experimental shading alters leaf litter breakdown in streams of contrasting riparian canopy cover

1. Headwater stream ecosystems are primarily heterotrophic, with allochthonous organic matter being the dominant energy. However, sunlight indirectly influences ecosystem structure and functioning, affecting microbial and invertebrate consumers and, ultimately, leaf litter breakdown. We tested the effects of artificial shading on litter breakdown rates in an open‐canopy stream (high ambient light) and a closed‐canopy stream (low ambient light). We further examined the responses of invertebrate shredders and aquatic hyphomycetes to shading to disentangle the underlying effects of light availability on litter breakdown. 2. Litter breakdown was substantially slower for both fast‐decomposing (alder, Alnus glutinosa) and slow‐decomposing (beech, Fagus sylvatica) leaf litters in artificially shaded stream reaches relative to control (no artificial shading) reaches, regardless of stream type (open or closed canopy). 3. Shredder densities were higher on A. glutinosa than on F. sylvatica litter, and shading had a greater effect on reducing shredder densities associated with A. glutinosa than those associated with F. sylvatica litter in both stream types. Fungal biomass was also negatively affected by shading. Results suggest that the effects of light availability on litter breakdown rates are mediated by resource quality and consumer density. 4. Results from feeding experiments, where A. glutinosa litter incubated under ambient light or artificial shade was offered to the shredder Gammarus fossarum, suggest that experimental shading and riparian canopy openness influenced litter palatability interactively. Rates of litter consumption by G. fossarum were decreased by experimental shading in the open‐canopy stream only. 5. The results suggest that even small variations in light availability in streams can mediate substantial within‐stream heterogeneity in litter breakdown. This study provides further evidence that changes in riparian vegetation, and thus light availability, influence organic matter processing in heterotrophic stream ecosystems through multiple trophic levels.     

Breakdown of tussock grass in streams along a gradient of agricultural development in New Zealand

Summary 1. We measured the breakdown rate of tussock grass in 12 New Zealand streams in catchments that provided a gradient of agricultural development. We also examined the microbial and invertebrate communities associated with decomposing tussock litter.
2. Pristine streams in the study had low concentrations of dissolved inorganic nitrogen (<10  μ g L⁻¹) and dissolved reactive phosphate (<3  μ g L⁻¹), whereas streams in the most developed catchments had high concentrations of nitrate (>2500  μ g L⁻¹) and phosphate (35  μ g L⁻¹), as well as greater amounts of suspended sediment and fine sediment covering the streambed.
3. Breakdown rate and microbial respiration were significantly related across the sites, and both were positively related to concentrations of nitrate and phosphate. Fungal biomass, measured as ergosterol, was positively related to microbial respiration and was also higher at sites with higher concentrations of nutrients. Total and shredding invertebrates were most abundant at the sites with high nutrient concentrations, but abundance of shredding invertebrates was not significantly related to breakdown rate. Amphipods were the most common shredding invertebrate at most sites, but probably did not contribute greatly to high rates of breakdown in streams in agricultural catchments.
4. With the exception of one site, nutrients from agricultural development appeared to have larger positive effects on litter breakdown than negative effects from sedimentation. Litter breakdown can serve as a functional measure of ecosystem health in streams, but caution should be exercised when a stress, such as land use, can have both positive (nutrients) and negative (sedimentation) effects.     

Changes in land plant function over the Phanerozoic: reconstructions based on the fossil record

Major fluctuations in the concentrations of atmospheric CO₂ and O₂, are predicted by historical long-term carbon and oxygen cycle models of atmospheric evolution and will have impacted directly on past climates, plant function and evolutionary processes. Here, palaeobotanical evidence is presented from the stomatal density record of fossil leaves spanning the past 400 Myr supporting the predicted changes in atsmopheric CO₂. Evidence from experiments on plants exposed to long-term high CO₂, environments and the newly-assembled fossil data indicate the potential for genetic modification of stomatal characters. The influence of the changes in fossil stomatal characteristics and atmospheric composition on the rates of leaf gas exchange over the course of land plant evolution has been investigated through modelling. Three contrasting eras of plain water economics emerge in the Devonian (high), Carboniferous (low) and from the Upper Jurassic to the present-day (high but declining). These patterns of change result from structural changes of the leaves and the impact of atmospheric CO₂, and O₂, concentrations on RuBisCo function and are consistent with the fossil evidence of sequential appearances of novel plant anatomical changes. The modelling approach is tested by comparing predicted leaf stable carbon isotope ratios with those measured on fossil plant and organic material. Viewed in a geological context, current and future increases in the concentration of atmospheric CO₂, might be considered as restoring plant function to that more typically experienced by plants over the majority of their evolutionary history.     

Microbial leaf degraders in boreal streams: bringing together stochastic and deterministic regulators of community composition

1. Leaves that fall into the water represent a new habitat for microorganisms to colonise in streams, providing an opportunity to study colonisation and the subsequent regulation of community structure. We explored community composition of bacteria and fungi on decomposing alder leaves in nine streams in central Sweden, and describe their relationship with environmental variables. Succession of the microbial community was studied in one of the streams for 118 days. Microbial community composition was examined by denaturing gradient gel electrophoresis on replicate samples of leaves from each stream. 2. During succession in one stream, maximum taxon richness was reached after 34 days for bacteria and 20 days for fungi respectively. Replicate samples within this stream differed between each other earlier in colonisation, while subsequently such variation among replicate communities was low and remained stable for several weeks. Replicate samples taken from all the nine streams after 34 days of succession showed striking similarities in microbial communities within‐streams, although communities differed more strongly between streams. 3. Canonical analysis of microbial communities and environmental variables revealed that water chemistry had a significant influence on community composition. This influence was superimposed on a statistical relationship between the properties of stream catchments and microbial community composition. 4. The catchment regulates microbial communities in two different ways. It harbours the species pool from which the in‐stream microbial community is drawn and it governs stream chemistry and the composition of organic substrates that further shape the communities. We suggest that there is a random element to colonisation early in succession, whereas other factors such as species interactions, stream chemistry and organic substrate properties, result in a more deterministic regulation of communities during later stages.     

Interactions between atmospheric CO2 enrichment and soil fauna

We have reviewed the responses of soil fauna to increased concentrations of atmospheric CO₂ and the consequent climate change. These will affect several attributes of animal populations and communities including their density, biomass, diversity, activity, rates of consumption, life history parameters and migration ability. Changes in the quality and quantity of litter and global warming are the main factors which are expected to modify soil fauna. Although changes have been observed in several attributes of the soil fauna as a consequence of increased concentrations of atmospheric CO₂, no general trend which might allow to the prediction of a general pattern of response has been identified. Because of the complexity of the biological mechanisms and the synergetic action of several factors, the few resulting responses reported in the literature are inconclusive. However, some aspects of the situation deserve more attention. These include the consequences of (1) changes in the food resources for soil fauna in the litter layer and in the rhizosphere, (2) the consumption of low quality litter by the macrofauna, (3) the change in life span in response to temperature elevation, (4) the enhancement of earthworm burrowing activity and (5) the changes in community composition arising because of specific differential resistance to adverse conditions. This revised version was published online in June 2006 with corrections to the Cover Date.     

The effects of low pH and palliative liming on beech litter decomposition in acid-sensitive streams

The decomposition of allochthonous leaf litter is retarded by stream acidification, but few studies have evaluated whether this effect can be offset by liming – the palliative addition of calcium carbonate either to streams or their catchments. We assessed the response of litter decomposition to pH and experimental liming in Welsh upland streams. Small-mesh (<335 μm) litter-bags containing common beech (Fagus sylvatica L.) were submerged in main river sites along the River Wye, and in replicate acid, circumneutral and experimentally limed tributaries (all n = 3) for 20 days. Beech decomposition was inhibited in acid tributaries and main river sites compared to circumneutral tributaries. Despite having only moderately increased pH relative to acid streams, limed sites had increased decomposition rates that were indistinguishable from naturally circumneutral streams. Decomposition rates increased highly significantly with pH across all 12 sites studied, and values were near identical to those in more prolonged experiments elsewhere. There were no significant variations in shredder numbers with decomposition rate, and no evidence that sites with faster decomposition had smaller shredder proportions. Although based on short-term observations and leaves from just one tree species, these results are consistent with the well-known retardation at low pH of some aspect microbial decomposition (e.g. by hyphomycete fungi). They are among the first to suggest that stream liming to combat acidification might reverse such impacts of low pH. Further data are required on the microbiological causes and ecological consequences of altered detrital processing in acid-sensitive and limed streams.     

Bayesian correlation between temperature and blossom onset data

The recent quantification of changes in time series of phenology data with Bayesian methods has provided compelling evidence for changes during the last 20 years. In this paper we correlate the phenological observations with spring temperature time series. We provide quantitative answers to the question whether changes in temperature and phenological time series should be regarded as coherent or independent. For the three considered species snowdrops, cherry and lime tree we find factors of 1.05, 2.19 and 3.26, respectively, in favor of coherence. The functional behavior and the trend in the temperature time series are presented. They amount to 0.15°C yr^?1 for the January–March average, 0.09°C yr^?1 for February–April and 0.1°C yr^?1 for March–May in 2002. In addition, we compare blossom trends for the coherent and independent hypotheses and find that the transition from trend values slightly positive before 1970 to strongly negative at present becomes sharper as the temperature data are included in the analysis.     

The fate of assimilated nitrogen in streams: an in situ benthic chamber study

1. Nitrogen (N) processing in streams has been investigated using whole‐stream ¹⁵N addition experiments that, in general, have found that a large proportion of added nitrate removed from the water column appears to be assimilated by the stream benthos. The long‐term fate of this retained N is unknown, and of particular interest is the possibility that it becomes denitrified through coupled mineralisation–nitrification–denitrification processes (indirect denitrification). 2. We used in situ chambers to produce highly ¹⁵N‐enriched benthic biofilms and removed the chambers to allow biofilms to interact with ambient stream conditions. Nitrogen assimilation and direct denitrification were estimated from the first chamber deployment. Chambers were periodically reinstalled over 4 weeks to measure tracer ¹⁵N in ammonium (), nitrate () and dinitrogen (N₂), from which we estimated subsequent rates of biotic N transformations, including N mineralisation (ammonification), nitrification and indirect denitrification. We also estimated rates of depuration of ¹⁵N tracer from benthic biomass compartments. 3. Nitrate uptake was roughly equivalent in the sand and cobble habitats that dominated the stream. Direct denitrification (denitrification of from the water column) was an order of magnitude higher in cobble habitats than in sand habitats, accounting for c. 26 and 2% of total nitrate uptake in cobble and sand, respectively. 4. Mean residence times of actively cycling organic N in stream benthos (algae and microbes) were 16 days in cobble habitats and 9 days in sand habitats. The difference between habitat types was driven by the influence of N residence time in epilithic biofilms (18 days) on cobbles. 5. Release of enriched ¹⁵ was the primary flux of remineralised N, while release of enriched ¹⁵ was an order of magnitude less. We detected slight ¹⁵N enrichment in dissolved nitrogen gas (N₂) in post‐enrichment sampling, indicating that indirect denitrification was taking place. However, indirect denitrification accounted for <0.1% of the assimilated N. 6. These experiments agree with results of whole‐stream ¹⁵N additions, in that most added N was assimilated rather than directly denitrified. Assimilation was primarily a short‐term N retention mechanism in this stream, and indirect denitrification of assimilated N accounted for only a minor proportion of the observed ¹⁵N loss over time. 7. Remaining possible fates include export of N as particulate organic matter, which may lead to additional storage of assimilated N in downstream habitats, and consumption by grazers.