, Volume 108, Issue 1–3, pp 39–54 | Cite as

Denitrification kinetics and denitrifier abundances in sediments of lakes receiving atmospheric nitrogen deposition (Colorado, USA)



The transport and deposition of anthropogenic nitrogen (N) to downwind ecosystems is significant and can be a dominant source of new N to many watersheds. Bacterially mediated denitrification in lake sediments may ameliorate the effects of N loading by permanently removing such inputs. We measured denitrification in sediments collected from lakes in the Colorado Rocky Mountains (USA) receiving elevated (5–8 kg N ha−1 y−1) or low (<2 kg N ha−1 y−1) inputs of atmospheric N deposition. The nitrate (NO3 ) concentration was significantly greater in high-deposition lakes (11.3 μmol l−1) compared to low-deposition lakes (3.3 μmol l−1). Background denitrification was positively related to NO3 concentrations and we estimate that the sampled lakes are capable of removing a significant portion of N inputs via sediment denitrification. We also conducted a dose–response experiment to determine whether chronic N loading has altered sediment denitrification capacity. Under Michaelis–Menten kinetics, the maximum denitrification rate and half-saturation NO3 concentration did not differ between deposition regions and were 765 μmol N m−2 h−1 and 293 μmol l−1 NO3 , respectively, for all lakes. We enumerated the abundances of nitrate- and nitrite-reducing bacteria and found no difference between high- and low-deposition lakes. The abundance of these bacteria was related to available light and bulk sediment resources. Our findings support a growing body of evidence that lakes play an important role in N removal and, furthermore, suggest that current levels of N deposition have not altered the abundance of denitrifying bacteria or saturated the capacity for sediment denitrification.


Atmospheric nitrogen deposition Denitrification Lake Denitrifier abundance Sediment 



We thank Marcia Kyle, Cathy Kochert, Erin Seybold, and Melanie Engstrom for their assistance with field and laboratory work. We are also grateful to the staff of the Mountain Research Station, Niwot Ridge Long Term Ecological Research Program, Rocky Mountain Biological Laboratory, Mountain Studies Institute, and Rocky Mountain National Park for their contributions in facilitating this study. Research funding was provided by National Science Foundation grant DEB-0516494 to JJE and graduate student research grants to MLM from the Mountain Studies Institute, the American Alpine Association, and the Society of Wetland Scientists. The suggestions of two anonymous referees greatly improved this manuscript.


  1. Aber J, McDowell W, Nadelhoffer K, Magill A, Berntson G, Kamakea M, McNulty S, Currie W, Rustad L, Fernandez I (1998) Nitrogen saturation in temperate forests; hypotheses revised. Bioscience 48:921–934CrossRefGoogle Scholar
  2. Baron JS, Campbell DH (1997) Nitrogen fluxes in a high elevation Colorado Rocky Mountain basin. Hydrol Process 11:783–799CrossRefGoogle Scholar
  3. Baron JS, Rueth HM, Wolfe AM, Nydick KR, Allstott EJ, Minear JT, Moraska B (2000) Ecosystem responses to nitrogen deposition in the Colorado Front Range. Ecosystems 3:352–368CrossRefGoogle Scholar
  4. Bergstrom AK, Jansson M (2006) Atmospheric nitrogen deposition has caused nitrogen enrichment and eutrophication of lakes in the northern hemisphere. Glob Change Biol 12:635–643CrossRefGoogle Scholar
  5. Bernot MJ, Dodds WK (2005) Nitrogen retention, removal, and saturation in lotic ecosystems. Ecosystems 8:442–453CrossRefGoogle Scholar
  6. Betlach MR, Tiedje JM (1981) Kinetic explanation for accumulation of nitrite, nitric-oxide, and nitrous-oxide during bacterial denitrification. Appl Environ Microbiol 42:1074–1084Google Scholar
  7. Bowden RD, Davidson E, Savage K, Arabia C, Steudler P (2004) Chronic nitrogen additions reduce total soil respiration and microbial respiration in temperate forest soils at the Harvard Forest. For Ecol Manag 196:43–56CrossRefGoogle Scholar
  8. Burns DA (2004) The effects of atmospheric nitrogen deposition in the Rocky Mountains of Colorado and southern Wyoming, USA—a critical review. Environ Pollut 127:257–269CrossRefGoogle Scholar
  9. Campbell DH, Kendall C, Chang CCY, Silva SR, Tonnessen KA (2002) Pathways for nitrate release from an alpine watershed: determination using delta N-15 and delta O-18. Water Resour Res 38:9Google Scholar
  10. Cannavo P, Richaume A, Lafolie F (2004) Fate of nitrogen and carbon in the vadose zone: in situ and laboratory measurements of seasonal variations in aerobic respiratory and denitrifying activities. Soil Biol Biochem 36:463–478CrossRefGoogle Scholar
  11. Cleveland CC, Townsend AR, Schmidt SK (2002) Phosphorus limitation of microbial processes in moist tropical forests: evidence from short-term laboratory incubations and field studies. Ecosystems 5:680–691Google Scholar
  12. Clow DW, Sickman JO, Striegl RG, Krabbenhoft DP, Elliott JG, Dornblaser M, Roth DA, Campbell DH (2003) Changes in the chemistry of lakes and precipitation in high-elevation national parks in the western United States, 1985–1999. Water Resour Res 39:1171CrossRefGoogle Scholar
  13. Cotner JB, Biddanda BA (2002) Small players, large role: microbial influence on biogeochemical processes in pelagic aquatic ecosystems. Ecosystems 5:105–121CrossRefGoogle Scholar
  14. Dillon PJ, Molot LA (1990) The role of ammonium and nitrate retention in the acidification of lakes and forested catchments. Biogeochemistry 11:23–43CrossRefGoogle Scholar
  15. Dong LF, Thornton CO, Nedwell DB, Underwood GJC (2000) Denitrification in sediments of the River Colne estuary, England. Mar Eco Progr Ser 203:109–122CrossRefGoogle Scholar
  16. Earl SR, Valett HM, Webster JR (2006) Nitrogen saturation in stream ecosystems. Ecology 87:3140–3151CrossRefGoogle Scholar
  17. Eisenlord SD, Zak DR (2010) Simulated atmospheric nitrogen deposition alters actinobacterial community composition in forest soils. Soil Sci Soc Am J 74:1157–1166CrossRefGoogle Scholar
  18. Elser JJ, Andersen T, Baron JS, Bergstrom AK, Jansson M, Kyle M, Nydick KR, Steger L, Hessen DO (2009a) Shifts in lake N:P stoichiometry and nutrient limitation driven by atmospheric nitrogen deposition. Science 326:835–837CrossRefGoogle Scholar
  19. Elser JJ, Kyle M, Steger L, Nydick KR, Baron JS (2009b) Nutrient availability and phytoplankton nutrient limitation across a gradient of atmospheric nitrogen deposition. Ecology 90:3062–3073CrossRefGoogle Scholar
  20. Elser JJ, Peace A, Kyle M, Wojewodzic M, McCrackin ML, Andersen T, Hessen DO (2010) Atmospheric nitrogen deposition is associated with elevated phosphorus limitation of lake zooplankton. Ecol Lett 13:1256–1261CrossRefGoogle Scholar
  21. Fisk MC, Schmidt SK (1995) Nitrogen mineralization and microbial biomass nitrogen dynamics in 3 alpine tundra communities. Soil Sci Soc Am J 59:1036–1043CrossRefGoogle Scholar
  22. Galloway JN, Aber JD, Erisman JW, Seitzinger SP, Howarth RW, Cowling EB, Cosby BJ (2003) The nitrogen cascade. Bioscience 53:341–356CrossRefGoogle Scholar
  23. Galloway JN, Dentener FJ, Capone DG, Boyer EW, Howarth RW, Seitzinger SP, Asner GP, Cleveland CC, Green PA, Holland EA, Karl DM, Michaels AF, Porter JH, Townsend AR, Vorosmarty CJ (2004) Nitrogen cycles: past, present, and future. Biogeochemistry 70:153–226CrossRefGoogle Scholar
  24. Groffman PM, Altabet MA, Bohlke JK, Butterbach-Bahl K, David MB, Firestone MK, Giblin AE, Kana TM, Nielsen LP, Voytek MA (2006) Methods for measuring denitrification: Diverse approaches to a difficult problem. Ecol Appl 16:2091–2122CrossRefGoogle Scholar
  25. Håkanson L, Jansson M (2002) Principles of Lake Sedimentology. Blackburn Press, CaldwellGoogle Scholar
  26. Harrison JA, Maranger RJ, Alexander RB, Giblin AE, Jacinthe PA, Mayorga E, Seitzinger SP, Sobota DJ, Wollheim WM (2009) The regional and global significance of nitrogen removal in lakes and reservoirs. Biogeochemistry 93:143–157CrossRefGoogle Scholar
  27. Hasegawa T, Okino T (2004) Seasonal variation of denitrification rate in Lake Suwa sediment. Limnology 5:33–39CrossRefGoogle Scholar
  28. Johnson SL, Neuer S, Garcia-Pichel F (2007) Export of nitrogenous compounds due to incomplete cycling within biological soil crusts of arid lands. Environ Microbiol 9:680–689CrossRefGoogle Scholar
  29. Kelly CA, Rudd JWM, Hesslein RH, Schindler DW, Dillon CT, Driscoll CT, Gherini SA, Hecky RE (1987) Prediction of biological acid neutralization in acid-sensitive lakes. Biogeochemistry 3:129–140CrossRefGoogle Scholar
  30. Kent HC, Porter KW (eds) (1980) Colorado geology. Rocky Mountain Association of Geologists, DenverGoogle Scholar
  31. Knowles R (1982) Denitrification. Microbiol Rev 46:43–70Google Scholar
  32. Koike I, Hattori A (1978) Denitrification and ammonia formation in anaerobic coastal sediments. Appl Environ Microbiol 35:278–282 Google Scholar
  33. Laverman AM, Van Cappellen P, van Rotterdam-Los D, Pallud C, Abell J (2006) Potential rates and pathways of microbial nitrate reduction in coastal sediments. FEMS Microbiol Ecol 58:179–192CrossRefGoogle Scholar
  34. Lovett GM, Rueth HM (1999) Soil nitrogen transformations in beech and maple stands along a nitrogen deposition gradient. Ecol Appl 9:1330–1344CrossRefGoogle Scholar
  35. Lukkari K, Hartikainen H, Leivuori M (2007) Fractionation of sediment phosphorus revisited. I: Fractionation steps and their biogeochemical basis. Limnol Oceanogr Methods 5:433–444CrossRefGoogle Scholar
  36. McCarty GW, Mookherji S, Angier JT (2007) Characterization of denitrification activity in zones of groundwater exfiltration within a riparian wetland ecosystem. Biol Fertil Soils 43:691–698CrossRefGoogle Scholar
  37. McCrackin ML, Elser JJ (2010) Atmospheric nitrogen deposition alters denitrification and nitrous oxide production in lake sediments. Ecology 91:528–539CrossRefGoogle Scholar
  38. Molot LA, Dillon PJ (1993) Nitrogen mass balances and denitrification rates in central Ontario lakes. Biogeochemistry 20:195–212CrossRefGoogle Scholar
  39. Mulholland PJ, Helton AM, Poole GC, Hall RO, Hamilton SK, Peterson BJ, Tank JL, Ashkenas LR, Cooper LW, Dahm CN, Dodds WK, Findlay SEG, Gregory SV, Grimm NB, Johnson SL, McDowell WH, Meyer JL, Valett HM, Webster JR, Arango CP, Beaulieu JJ, Bernot MJ, Burgin AJ, Crenshaw CL, Johnson LT, Niederlehner BR, O’Brien JM, Potter JD, Sheibley RW, Sobota DJ, Thomas SM (2008) Stream denitrification across biomes and its response to anthropogenic nitrate loading. Nature 452:202–206CrossRefGoogle Scholar
  40. Musselman RC, Slauson WL (2004) Water chemistry of high elevation Colorado wilderness lakes. Biogeochemistry 71:387–414CrossRefGoogle Scholar
  41. Nanus L, Campbell DH, Ingersoll GP, Clow DW, Mast MA (2003) Atmospheric deposition maps for the Rocky Mountains. Atmos Environ 37:4881–4892CrossRefGoogle Scholar
  42. Nanus L, Williams MW, Campbell DH, Elliott EM, Kendall C (2008) Evaluating regional patterns in nitrate sources to watersheds in national parks of the Rocky Mountains using nitrate isotopes. Environ Sci Technol 42:6487–6493CrossRefGoogle Scholar
  43. Nanus L, Williams MW, Campbell DH, Tonnessen KA, Blett T, Clow DW (2009) Assessment of lake sensitivity to acidic deposition in national parks of the Rocky Mountains. Ecol Appl 19:961–973CrossRefGoogle Scholar
  44. Nielsen K, Risgaard-Petersen N, Somod B, Rysgaard S, Bergo T (2001) Nitrogen and phosphorus retention estimated independently by flux measurements and dynamic modelling in the estuary, Randers Fjord, Denmark. Mar Ecol Progr Ser 219:25–40CrossRefGoogle Scholar
  45. O’Connor BL, Hondzo M, Dobraca D, LaPara TM, Finlay JC, Brezonik PL (2006) Quantity-activity relationship of denitrifying bacteria and environmental scaling in streams of a forested watershed. J Geophys Res Biogeosci 111:G04014CrossRefGoogle Scholar
  46. Oremland RS, Umberfer C, Culbertson CW, Smith RL (1984) Denitrification in San Francisco Bay intertidal sediments. Appl Environ Microbiol 47:1106–1112 Google Scholar
  47. Oren A, Blackburn TH (1979) Estimation of sediment denitrification rates at insitu nitrate concentrations. Appl Environ Microbiol 37:174–176 Google Scholar
  48. Pina-Ochoa E, Alvarez-Cobelas M (2006) Denitrification in aquatic environments: a cross-system analysis. Biogeochemistry 81:111–130CrossRefGoogle Scholar
  49. Richardson WB, Strauss EA, Bartsch LA, Monroe EM, Cavanaugh JC, Vingum L, Soballe DM (2004) Denitrification in the Upper Mississippi River: rates, controls, and contribution to nitrate flux. Can J Fish Aquat Sci 61:1102–1112CrossRefGoogle Scholar
  50. Rowe R, Todd R, Waide J (1977) Microtechnique for most-probable-number analysis. Appl Environ Microbiol 33:675–680Google Scholar
  51. Rudd JWM, Kelly CA, Schindler DW, Turner MA (1988) Disruption of the nitrogen-cycle in acidified lakes. Science 240:1515–1517CrossRefGoogle Scholar
  52. Saunders DL, Kalff J (2001) Nitrogen retention in wetlands, lakes and rivers. Hydrobiologia 443:205–212CrossRefGoogle Scholar
  53. Seitzinger SP, Harrison JA, Bohlke JK, Bouwman AF, Lowrance R, Peterson B, Tobias C (2006) Denitrification across landscapes and waterscapes: a synthesis. Ecol Appl 16:2064–2090CrossRefGoogle Scholar
  54. Sharma S, Szele Z, Schilling R, Munch JC, Schloter M (2006) Influence of freeze-thaw stress on the structure and function of microbial communities and denitrifying populations in soil. Appl Environ Microbiol 72:2148–2154CrossRefGoogle Scholar
  55. Sickman JO, Melack JM, Stoddard JL (2002) Regional analysis of inorganic nitrogen yield and retention in high-elevation ecosystems of the Sierra Nevada and Rocky Mountains. Biogeochemistry 57:341–374CrossRefGoogle Scholar
  56. Silvennoinen H, Liikanen A, Torssonen J, Stange CF, Martikainen PJ (2008) Denitrification and nitrous oxide effluxes in boreal, eutrophic river sediments under increasing nitrate load: a laboratory microcosm study. Biogeochemistry 91:105–116CrossRefGoogle Scholar
  57. Sotomayor D, Rice CW (1996) Denitrification in soil profiles beneath grassland and cultivated soils. Soil Sci Soc Am J 60:1822–1828CrossRefGoogle Scholar
  58. Staley TE, Griffin JB (1981) Simultaneous enumeration of denitrifying and nitrate reducing bacteria by a microtiter most-probable-number (MPN) procedure. Soil Biol Biochem 13:385–388CrossRefGoogle Scholar
  59. Stoddard JL (1994) Long-term changes in watershed retention of nitrogen—its causes and aquatic consequences. In: Baker LA (ed) Environmental chemistry of lakes and reservoirs. Amer Chemical Soc, Washington, DC, pp 223–284CrossRefGoogle Scholar
  60. Sundbäck K, Linares F, Larson F, Wulff A (2004) Benthic nitrogen fluxes along a depth gradient in a microtidal fjord: the role of denitrification and microphytobenthos. Limnol Oceanogr 49:1095–1107CrossRefGoogle Scholar
  61. Tietema A (1998) Microbial carbon and nitrogen dynamics in coniferous forest floor material collected along a European nitrogen deposition gradient. For Ecol Manag 101:29–36CrossRefGoogle Scholar
  62. Tørseth K, Semb A (1998) Deposition of nitrogen and other major inorganic compounds in Norway, 1992–1996. Environ Pollut 102:299–304CrossRefGoogle Scholar
  63. Treseder KK (2008) Nitrogen additions and microbial biomass: a meta-analysis of ecosystem studies. Ecol Lett 11:1111–1120CrossRefGoogle Scholar
  64. Wallenstein MD, McNulty S, Fernandez IJ, Boggs J, Schlesinger WH (2006) Nitrogen fertilization decreases forest soil fungal and bacterial biomass in three long-term experiments. For Ecol Manag 222:459–468CrossRefGoogle Scholar
  65. Wetzel RG (2001) Limnology: lake and river ecosystems. Academic Press, San DiegoGoogle Scholar
  66. Williams MW, Baron JS, Caine N, Sommerfeld R, Sanford R (1996) Nitrogen saturation in the Rocky Mountains. Environ Sci Technol 30:640–646CrossRefGoogle Scholar
  67. Wolfe AP, Cooke CA, Hobbs WO (2006) Are current rates of atmospheric nitrogen deposition influencing lakes in the eastern Canadian arctic? Arct Antarct Alp Res 38:465–476CrossRefGoogle Scholar
  68. Wollheim WM, Vorosmarty CJ (2006) Relationship between river size and nutrient removal. Geophys Res Lett 33:L06410. doi: 06410.01029/02006GL025845 CrossRefGoogle Scholar
  69. Wollheim WM, Vorosmarty CJ, Bouwman AF, Green P, Harrison J, Linder E, Peterson BJ, Seitzinger SP, Syvitski JPM (2008) Global N removal by freshwater aquatic systems using a spatially distributed, within-basin approach. Global Biogeochem Cycles 22:Gb2026. doi: 2010.1029/2007GB002963 CrossRefGoogle Scholar
  70. Yoshinari T, Knowles R (1976) Acetylene inhibition of nitrous oxide reduction by denitrifying bacteria. Biochem Biophys Res Commun 69:705–710CrossRefGoogle Scholar
  71. Yu KW, DeLaune RD, Boeckx P (2006) Direct measurement of denitrification activity in a Gulf coast freshwater marsh receiving diverted Mississippi River water. Chemosphere 65:2449–2455 CrossRefGoogle Scholar
  72. Zak DR, Holmes WE, Tomlinson MJ, Pregitzer KS, Burton AJ (2006) Microbial cycling of C and N in northern hardwood forests receiving chronic atmospheric NO3 deposition. Ecosystems 9:242–253CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  1. 1.School of Life SciencesArizona State UniversityTempeUSA

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