Anthropogenic addition of bioavailable nitrogen to the biosphere is increasing1,2 and terrestrial ecosystems are becoming increasingly nitrogen-saturated3, causing more bioavailable nitrogen to enter groundwater and surface waters4,5,6. Large-scale nitrogen budgets show that an average of about 20–25 per cent of the nitrogen added to the biosphere is exported from rivers to the ocean or inland basins7,8, indicating that substantial sinks for nitrogen must exist in the landscape9. Streams and rivers may themselves be important sinks for bioavailable nitrogen owing to their hydrological connections with terrestrial systems, high rates of biological activity, and streambed sediment environments that favour microbial denitrification6,10,11. Here we present data from nitrogen stable isotope tracer experiments across 72 streams and 8 regions representing several biomes. We show that total biotic uptake and denitrification of nitrate increase with stream nitrate concentration, but that the efficiency of biotic uptake and denitrification declines as concentration increases, reducing the proportion of in-stream nitrate that is removed from transport. Our data suggest that the total uptake of nitrate is related to ecosystem photosynthesis and that denitrification is related to ecosystem respiration. In addition, we use a stream network model to demonstrate that excess nitrate in streams elicits a disproportionate increase in the fraction of nitrate that is exported to receiving waters and reduces the relative role of small versus large streams as nitrate sinks.

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  1. 1.

    et al. Human alteration of the global nitrogen cycle: Sources and consequences. Ecol. Appl. 7, 737–750 (1997)

  2. 2.

    et al. Nitrogen cycles past, present, and future. Biogeochemistry 70, 153–226 (2004)

  3. 3.

    et al. Nitrogen saturation in temperate forest ecosystems. Bioscience 48, 921–934 (1998)

  4. 4.

    , , , & Exploring changes in river nitrogen export to the world’s oceans. Glob. Biogeochem. Cycles 19 GB1002 10.1029/2004GB002314 (2005)

  5. 5.

    & Landscape, regional and global estimates of nitrogen flux from land to sea: Errors and uncertainties. Biogeochemistry 57, 429–476 (2002)

  6. 6.

    , & Global change: The nitrogen cycle and rivers. Water Resource Res. 42 W03S06 10.1029/2005WR004300 (2006)

  7. 7.

    et al. Riverine inputs of nitrogen to the North Atlantic Ocean: fluxes and human influences. Biogeochemistry 35, 75–139 (1996)

  8. 8.

    et al. Riverine nitrogen export from the continents to the coasts. Glob. Biogeochem. Cycles 20 GB1S91 10.1029/2005GB002537 (2006)

  9. 9.

    et al. Where did all the nitrogen go? Fate of nitrogen inputs to large watersheds in the northeastern USA. Biogeochemistry 57, 267–293 (2002)

  10. 10.

    et al. Denitrification across landscapes and waterscapes: a synthesis. Ecol. Appl. 16, 2064–2090 (2006)

  11. 11.

    et al. Thermodynamic constraints on nitrogen transformations and other biogeochemical processes at soil–stream interfaces. Ecology 79, 684–703 (1998)

  12. 12.

    Stream Solute Workshop. Concepts and methods for assessing solute dynamics in stream ecosystems. J. N. Am. Benthol. Soc. 9, 95–119 (1990)

  13. 13.

    et al. N uptake as a function of concentration in streams. J. N. Am. Benthol. Soc. 21, 206–220 (2002)

  14. 14.

    , , , & The saturation of N cycling in Central Plains streams: 15N experiments across a broad gradient of nitrate concentrations. Biogeochemistry 10.1007/s10533–007–9073–7 (2007)

  15. 15.

    & Ecosystem metabolism controls nitrogen uptake in streams in Grand Teton National Park, Wyoming. Limnol. Oceanogr. 48, 1120–1128 (2003)

  16. 16.

    et al. Effects of light on NO3- uptake in small forested streams: diurnal and day-to-day variations. J. N. Am. Benthol. Soc. 25, 583–595 (2006)

  17. 17.

    , , & Denitrification in nitrate-rich streams: Diurnal and seasonal variation related to benthic oxygen metabolism. Limnol. Oceanogr. 35, 640–651 (1990)

  18. 18.

    & Denitrification in aquatic environments: A cross-system analysis. Biogeochemistry 81, 111–130 (2006)

  19. 19.

    , & Effect of stream channel size on the delivery of nitrogen to the Gulf of Mexico. Nature 403, 758–761 (2000)

  20. 20.

    et al. Control of nitrogen export from watersheds by headwater streams. Science 292, 86–90 (2001)

  21. 21.

    et al. Nitrogen retention in rivers: Model development and application to watersheds in the northeastern USA. Biogeochemistry 57, 199–237 (2002)

  22. 22.

    , , , & Relationship between river size and nutrient removal. Geophys. Res. Lett. 33, L06410 (2006)

  23. 23.

    , , & Nitrate flux in the Mississippi River. Nature 414, 166–167 (2001)

  24. 24.

    , , , & Environmental, economic, and energetic costs and benefits of biodiesel and ethanol biofuels. Proc. Natl Acad. Sci. USA 103, 11206–11210 (2006)

  25. 25.

    , , , & Agricultural sustainability and intensive production practices. Nature 418, 671–677 (2002)

  26. 26.

    et al. Global consequences of land use. Science 309, 570–574 (2005)

  27. 27.

    , & Agricultural runoff fuels large phytoplankton blooms in vulnerable areas of the ocean. Nature 434, 211–214 (2005)

  28. 28.

    , , & Measuring nutrient spiraling in streams. Can. J. Fish. Aquat. Sci. 38, 860–863 (1981)

  29. 29.

    & in Methods in Stream Ecology (eds Hauer, F. R. & Lamberti, G. A.) 169–186 (Elsevier, New York, 2006)

  30. 30.

    et al. Stream denitrification and total nitrate uptake rates measured using a field 15N isotope tracer approach. Limnol. Oceanogr. 49, 809–820 (2004)

  31. 31.

    et al. Natural abundance-level measurement of the nitrogen isotopic composition of oceanic nitrate: an adaptation of the ammonia diffusion method. Mar. Chem. 57, 227–242 (1997)

  32. 32.

    & Measurement of the stable isotope ratio of dissolved N2 in 15N tracer experiments. Limnol. Oceanogr. Methods 5, 233–240 (2007)

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Funding for this research was provided by the National Science Foundation. We thank N. Ostrom for assistance with stable isotope measurements of N2 and N2O, and W. Wollheim for initial development of the model that we modified to estimate denitrification rates from field data. We thank M. Mitchell, B. Roberts and E. Bernhardt for their comments on earlier versions of the paper. We thank the NSF LTER network, US Forest Service, National Park Service and many private landowners for permission to conduct experiments on their lands. Partial support to P.J.M. during manuscript preparation was provided by the US Department of Energy, Office of Science, Biological and Environmental Research under contract with UT-Battelle.

Author Contributions P.J.M. coordinated the stream 15N experiments and analysed the compiled experimental data sets. A.M.H. and G.C.P conducted the stream network modelling. P.J.M., A.M.H. and G.C.P. wrote major portions of the manuscript. S.K.H. established sampling protocols and coordinated the 15N analysis of dissolved N2 samples. Except for A.M.H., all authors listed to J.R.W. were joint project Principal Investigators and contributed to the conceptual and methodological development of the project and analysis of data. Authors listed from C.P.A. to S.M.T. coordinated field experiments and analysed data from one or more biomes. All authors discussed the results and commented on the manuscript.

Author information

Author notes

    • Jake J. Beaulieu
    • , Richard W. Sheibley
    •  & Daniel J. Sobota

    Present addresses: US Environmental Protection Agency, Cincinnati, Ohio 45268, USA (J.J.B.); US Geological Survey, Tacoma, Washington 98402, USA (R.W.S.); School of Earth and Environmental Sciences, Washington State University, Vancouver Campus, Vancouver, Washington 98686, USA (D.J.S.).


  1. Environmental Sciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831, USA

    • Patrick J. Mulholland
  2. Department of Ecology and Evolutionary Biology, University of Tennessee, Knoxville, Tennessee 37996, USA

    • Patrick J. Mulholland
    •  & Lee W. Cooper
  3. Odum School of Ecology, University of Georgia, Athens, Georgia 30602, USA

    • Ashley M. Helton
    • , Geoffrey C. Poole
    •  & Judy L. Meyer
  4. Eco-metrics, Inc., Tucker, Georgia 30084, USA

    • Geoffrey C. Poole
  5. Department of Zoology and Physiology, University of Wyoming, Laramie, Wyoming 82071, USA

    • Robert O. Hall
  6. Kellogg Biological Station, Michigan State University, Hickory Corners, Michigan 49060, USA

    • Stephen K. Hamilton
    • , Amy J. Burgin
    •  & Jonathan M. O’Brien
  7. Ecosystems Center, Marine Biological Laboratory, Woods Hole, Massachusetts 02543, USA

    • Bruce J. Peterson
    •  & Suzanne M. Thomas
  8. Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA

    • Jennifer L. Tank
    • , Clay P. Arango
    • , Jake J. Beaulieu
    •  & Laura T. Johnson
  9. Department of Fisheries and Wildlife, Oregon State University, Corvallis, Oregon 97331, USA

    • Linda R. Ashkenas
    • , Stanley V. Gregory
    •  & Daniel J. Sobota
  10. Department of Biology, University of New Mexico, Albuquerque, New Mexico 87131, USA

    • Clifford N. Dahm
    •  & Chelsea L. Crenshaw
  11. Division of Biology, Kansas State University, Manhattan, Kansas 66506, USA

    • Walter K. Dodds
  12. Institute of Ecosystem Studies, Millbrook, New York 12545, USA

    • Stuart E. G. Findlay
  13. School of Life Sciences, Arizona State University, Tempe, Arizona 85287, USA

    • Nancy B. Grimm
    •  & Richard W. Sheibley
  14. Pacific Northwest Research Station, US Forest Service, Corvallis, Oregon 97331, USA

    • Sherri L. Johnson
  15. Department of Natural Resources, University of New Hampshire, Durham, New Hampshire 03824, USA

    • William H. McDowell
    •  & Jody D. Potter
  16. Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia 24061, USA

    • H. Maurice Valett
    • , Jackson R. Webster
    •  & B. R. Niederlehner
  17. Department of Biology, Ball State University, Muncie, Indiana 47306, USA

    • Melody J. Bernot


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Corresponding author

Correspondence to Patrick J. Mulholland.

Supplementary information

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  1. 1.

    Supplementary Information

    This file contains Supplementary Methods and Supplementary Discussion with additional references; Supplementary Figures and Legends 1-3; and Supplementary Tables 1-3. The Supplementary Methods include more detailed 15N experiment methods and stream network model development. The Supplementary Discussion presents NO3- removal over a standardized stream reach, effects of land use and stream size on uptake rates, and limitations of the data and stream network modeling results. The Supplemental Figure 1 shows study site locations, Figure 2 shows the stream network model structure, and Figure 3 shows the location and hydrography of the Little Tennessee River network used in model simulations. The Supplementary Table 1 presents physical and chemical characteristics and NO3- uptake and denitrification rates determined for all streams in the 15N experiments, Table 2 includes definitions of model terms, and Table 3 presents the methods used to derive model parameters.

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