Quantifying Effluent Dissolved Organic Nitrogen (EDON) Uptake by Microbial Communities Along a Salinity Gradient in the York River
Effluent discharged from water reclamation facilities (WRFs) contains dissolved organic nitrogen, termed effluent dissolved organic nitrogen (EDON), that subsequently enters coastal waterways. It is still unclear at what rate EDON can be taken up by microbial communities relative to other nitrogen (N) substrates. Bench-scale sequencing batch reactors (SBRs), used to mimic WRFs, were supplied with 15N-labeled ammonium (15NH4+) to produce 15N-labeled EDON (EDO15N) that was subsequently used to measure uptake rates along a salinity gradient of the York River, Virginia, USA, in the spring and summer. Although NH4+ dominated influent N pools, only a small fraction (4.1%) of EDON was produced from NH4+ microbial assimilation in biological treatment processes. When added as a short-term (4-h) tracer, the EDO15N was taken up by estuarine microbes at rates 0.01–0.434 μmol N L−1 h−1, which are similar to rates of NH4+ and nitrate uptake. When added to 48-h bioassays, EDON stimulated phytoplankton growth more at the lower salinity (0–8‰) sites (8.5–13.8 μg Chl a L−1) than at the higher salinity (20‰) site (up to 0.4 μmol Chl a L−1). The microbes in the 0.7–5 μm size fraction had significantly higher EDO15N uptake rates than the larger size fraction (e.g., > 5 μm, p < 0.05). Taken together with urea and amino acids, DON plays a more important role in N nutrition for microbes during the summer months. This study provides the first EDO15N uptake rates using EDO15N produced from 15NH4+ in SBRs, and the results provide conclusive evidence that organic N in effluent is biologically available to estuarine microbes.
KeywordsEffluent Dissolved organic nitrogen (DON) Uptake Microbial communities York River
We are grateful to anonymous reviewers for their insightful suggestions. This paper is Contribution No. 3824 of the Virginia Institute of Marine Science, College of William & Mary.
This study was supported by The Water Environment Research Foundation grant no. U1R11 to DAB and RES. XY was supported by the China Scholarship Council (CSC) (No. 201704910658).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no competing interests.
- Aquino, S.F., and D.C. Stuckey. 2003. Production of soluble microbial products (SMP) in anaerobic chemostats under nutrient deficiency. Journal of Environmental Engineering 129 (11): 1007–1014. https://doi.org/10.1061/(ASCE)0733-9372(2003)129:11(1007).CrossRefGoogle Scholar
- Bronk, D.A. 2002. Dynamics of DON, p. 153–247. In Biogeochemistry of marine dissolved organic matter, ed. D.A. Hansell and C.A. Carlson. Cambridge: Academic Press.Google Scholar
- Bronk, D.A., P.M. Glibert, T.C. Malone, S. Banahan, and E. Sahlsten. 1998. Inorganic and organic nitrogen cycling in Chesapeake Bay: Autotrophic versus heterotrophic processes and relationships to carbon flux. Aquatic Microbial Ecology 15: 177–189. https://doi.org/10.3354/ame015177.CrossRefGoogle Scholar
- Bronk, D.A., Q.N. Roberts, M.P. Sanderson, E.A. Canuel, P.G. Hatcher, R. Mesfioui, K.C. Filippino, M.R. Mulholland, and N.G. Love. 2010. Effluent organic nitrogen (EON): Bioavailability and photochemical and salinity-mediated release. Environmental Science & Technology 44 (15): 5830–5835. https://doi.org/10.1021/es101115g.CrossRefGoogle Scholar
- Bronk, D.A., L. Killberg-Thoreson, R.E. Sipler, M.R. Mulholland, Q.N. Roberts, P.W. Bernhardt, M. Garrett, J.M. O’Neil, and C.A. Heil. 2014. Nitrogen uptake and regeneration (ammonium regeneration, nitrification and photoproduction) in waters of the West Florida Shelf prone to blooms of Karenia brevis. Harmful Algae 38: 50–62. https://doi.org/10.1016/j.hal.2014.04.007.CrossRefGoogle Scholar
- Eom, H., D. Borgatti, H.W. Paerl, and C. Park. 2017. Formation of low-molecular-weight dissolved organic nitrogen in predenitrification biological nutrient removal systems and its impact on eutrophication in coastal waters. Environmental Science & Technology 51 (7): 3776–3783. https://doi.org/10.1021/acs.est.6b06576.CrossRefGoogle Scholar
- Fan, C., P.M. Glibert, and J.M. Burkholder. 2003. Characterization of the affinity for nitrogen, uptake kinetics, and environmental relationships for Prorocentrum minimum in natural blooms and laboratory cultures. Harmful Algae 2 (4): 283–299. https://doi.org/10.1016/S1568-9883(03)00047-7.CrossRefGoogle Scholar
- Fan, L., M.T. Brett, W. Jiang, and B. Li. 2017. Dissolved organic nitrogen recalcitrance and bioavailable nitrogen quantification for effluents from advanced nitrogen removal wastewater treatment facilities. Environmental Pollution 229: 255–263. https://doi.org/10.1016/j.envpol.2017.05.093.CrossRefGoogle Scholar
- Filippino, K.C., M.R. Mulholland, P.W. Bernhardt, G.E. Boneillo, R.E. Morse, M. Semcheski, H. Marshall, N.G. Love, Q. Roberts, and D.A. Bronk. 2011. The bioavailability of effluent-derived organic nitrogen along an estuarine salinity gradient. Estuaries and Coasts 34 (2): 269–280. https://doi.org/10.1007/s12237-010-9314-1.CrossRefGoogle Scholar
- Gagnon, R., M. Levasseur, A.M. Weise, J. Fauchot, P.G.C. Campbell, B.J. Weissenboeck, A. Merzouk, M. Gosselin, and B. Vigneault. 2005. Growth stimulation of Alexandrium Tamarense (dinophyceae) by humic substances from the Manicouagan river (eastern Canada). Journal of Phycology 41 (3): 489–497. https://doi.org/10.1111/j.1529-8817.2005.00077.x.CrossRefGoogle Scholar
- Galloway, J.N., A.R. Townsend, J.W. Erisman, M. Bekunda, Z. Cai, J.R. Freney, L.A. Martinelli, S.P. Seitzinger, and M.A. Sutton. 2008. Transformation of the nitrogen cycle: Recent trends, questions, and potential solutions. Science 320 (5878): 889–892. https://doi.org/10.1126/science.1136674.CrossRefGoogle Scholar
- Glibert, P.M., F.P. Wilkerson, R.C. Dugdale, J.A. Raven, C.L. Dupont, P.R. Leavitt, A.E. Parker, J.M. Burkholder, and T.M. Kana. 2016. Pluses and minuses of ammonium and nitrate uptake and assimilation by phytoplankton and implications for productivity and community composition, with emphasis on nitrogen-enriched conditions. Limnology and Oceanography 61 (1): 165–197. https://doi.org/10.1002/lno.10203.CrossRefGoogle Scholar
- Grady, C.P.L., G.T. Daigger, N.G. Love, and C.D. Filipe. 2011. Biological wastewater treatment. Boca Raton: CRC press.Google Scholar
- Hansell, D.A., P.M. Williams, and B.B. Ward. 1993. Measurements of DOC and DON in the Southern California Bight using oxidation by high temperature combustion. Deep Sea Research Part I: Oceanographic Research Papers 40 (2): 219–234. https://doi.org/10.1016/0967-0637(93)90001-J.CrossRefGoogle Scholar
- Heisler, J., P.M. Glibert, J.M. Burkholder, D.M. Anderson, W. Cochlan, W.C. Dennison, Q. Dortch, C.J. Gobler, C.A. Heil, E. Humphries, A. Lewitus, R. Magnien, H.G. Marshall, K. Sellner, D.A. Stockwell, D.K. Stoecker, and M. Suddleson. 2008. Eutrophication and harmful algal blooms: A scientific consensus. Harmful Algae 8 (1): 3–13. https://doi.org/10.1016/j.hal.2008.08.006.CrossRefGoogle Scholar
- Hu, H., H. Ma, L. Ding, J. Geng, K. Xu, H. Huang, Y. Zhang, and H. Ren. 2016. Concentration, composition, bioavailability, and N-nitrosodimethylamine formation potential of particulate and dissolved organic nitrogen in wastewater effluents: A comparative study. Science of the Total Environment 569-570: 1359–1368. https://doi.org/10.1016/j.scitotenv.2016.06.218.CrossRefGoogle Scholar
- Killberg-Thoreson, L.M. 2011. A tale of two blooms: Dynamics of nitrogen uptake by harmful algae in the Eastern Gulf of Mexico and York River, Virginia, USA. Doctor of Philosophy, The College of William and Mary.Google Scholar
- Kirchman, D.L., R.G. Keil, and P.A. Wheeler. 1989. The effect of amino acids on ammonium utilization and regeneration by heterotrophic bacteria in the subarctic Pacific. Deep Sea Research Part A. Oceanographic Research Papers 36 (11): 1763–1776. https://doi.org/10.1016/0198-0149(89)90071-X.CrossRefGoogle Scholar
- Koroleff, F. 1983. Determination of ammonium. In Methods of Seawater Analysis, ed. K. Grasshoff, M. Ehrhardt, and K. Kremling, 2nd ed., 150–157. Weinheim: Verlag Chemie.Google Scholar
- Liu, H., J. Jeong, H. Gray, S. Smith, and D.L. Sedlak. 2012. Algal uptake of hydrophobic and hydrophilic dissolved organic nitrogen in effluent from biological nutrient removal municipal wastewater treatment systems. Environmental Science & Technology 46 (2): 713–721. https://doi.org/10.1021/es203085y.CrossRefGoogle Scholar
- Mesfioui, R., N.G. Love, D.A. Bronk, M.R. Mulholland, and P.G. Hatcher. 2012. Reactivity and chemical characterization of effluent organic nitrogen from wastewater treatment plants determined by Fourier transform ion cyclotron resonance mass spectrometry. Water Research 46 (3): 622–634. https://doi.org/10.1016/j.watres.2011.11.022.CrossRefGoogle Scholar
- Parsons, T.R., Y. Maita, and C.M. Lalli. 1984. A manual of biological and chemical methods for seawater analysis. Oxford: Pergamon Press.Google Scholar
- Perminova, I.V., I.V. Dubinenkov, A.S. Kononikhin, A.I. Konstantinov, A.Y. Zherebker, M.A. Andzhushev, V.A. Lebedev, E. Bulygina, R.M. Holmes, Y.I. Kostyukevich, I.A. Popov, and E.N. Nikolaev. 2014. Molecular mapping of sorbent selectivities with respect to isolation of Arctic dissolved organic matter as measured by Fourier transform mass spectrometry. Environmental Science & Technology 48 (13): 7461–7468. https://doi.org/10.1021/es5015423.CrossRefGoogle Scholar
- Qin, C., H. Liu, L. Liu, S. Smith, D.L. Sedlak, and A.Z. Gu. 2015. Bioavailability and characterization of dissolved organic nitrogen and dissolved organic phosphorus in wastewater effluents. Science of the Total Environment 511: 47–53. https://doi.org/10.1016/j.scitotenv.2014.11.005.CrossRefGoogle Scholar
- Sattayatewa, C., K. Pagilla, P. Pitt, K. Selock, and T. Bruton. 2009. Organic nitrogen transformations in a 4-stage Bardenpho nitrogen removal plant and bioavailability/biodegradability of effluent DON. Water Research 43 (18): 4507–4516. https://doi.org/10.1016/j.watres.2009.07.030.CrossRefGoogle Scholar
- Seitzinger, S.P., J.A. Harrison, E. Dumont, A.H.W. Beusen, and A.F. Bouwman. 2005. Sources and delivery of carbon, nitrogen, and phosphorus to the coastal zone: An overview of Global Nutrient Export from Watersheds (NEWS) models and their application. Global Biogeochemical Cycles 19 (4): n/a–n/a. https://doi.org/10.1029/2005GB002606.CrossRefGoogle Scholar
- Sharp, J.H., R. Benner, L. Bennett, C.A. Carlson, R. Dow, and S.E. Fitzwater. 1993. Re-evaluation of high temperature combustion and chemical oxidation measurements of dissolved organic carbon in seawater. Limnology and Oceanography 38 (8): 1774–1782. https://doi.org/10.4319/lo.19220.127.116.114.CrossRefGoogle Scholar
- Sharp, J.H., A.Y. Beauregard, D. Burdige, G. Cauwet, S.E. Curless, R. Lauck, K. Nagel, H. Ogawa, A.E. Parker, O. Primm, M. Pujo-Pay, W.B. Savidge, S. Seitzinger, G. Spyres, and R. Styles. 2004. A direct instrument comparison for measurement of total dissolved nitrogen in seawater. Marine Chemistry 84 (3-4): 181–193. https://doi.org/10.1016/j.marchem.2003.07.003.CrossRefGoogle Scholar
- Stücheli, P.E., J. Niggemann, and C.J. Schubert. 2018. Comparison of different solid phase extraction sorbents for the qualitative assessment of dissolved organic nitrogen in freshwater samples using FT-ICR-MS. Journal of Limnology 77: 400–411. https://doi.org/10.4081/jlimnol.2018.1791.Google Scholar
- The Chesapeake Bay Resource Library. 2009. Special Report URL (last accessed 23 October 2011): http://www.chesapeakebay.net/status_nitrogensources.aspx?menuitem=19797.
- Vaquer-Sunyer, R., D.J. Conley, S. Muthusamy, M.V. Lindh, J. Pinhassi, and E.S. Kritzberg. 2015. Dissolved organic nitrogen inputs from wastewater treatment plant effluents increase responses of planktonic metabolic rates to warming. Environmental Science & Technology 49 (19): 11411–11420. https://doi.org/10.1021/acs.est.5b00674.CrossRefGoogle Scholar
- Varela, M.M., A. Bode, E. Fernández, N. Gónzalez, V. Kitidis, M. Varela, and E.M.S. Woodward. 2005. Nitrogen uptake and dissolved organic nitrogen release in planktonic communities characterised by phytoplankton size–structure in the Central Atlantic Ocean. Deep Sea Research Part I: Oceanographic Research Papers 52 (9): 1637–1661. https://doi.org/10.1016/j.dsr.2005.03.005.CrossRefGoogle Scholar
- Veuger, B., J.J. Middelburg, H.T.S. Boschker, J. Nieuwenhuize, P. van Rijswijk, E.J. Rochelle-Newall, and N. Navarro. 2004. Microbial uptake of dissolved organic and inorganic nitrogen in Randers Fjord. Estuarine, Coastal and Shelf Science 61 (3): 507–515. https://doi.org/10.1016/j.ecss.2004.06.014.CrossRefGoogle Scholar