Stable isotopes as an effective tool for N nutrient source identification in a heavily urbanized and agriculturally intensive tropical lowland basin

Abstract

We present the application of dual stable isotope analyses of NO₃ (δ¹⁵N-NO₃ and δ¹⁸O-NO₃) to provide a comprehensive assessment of the provenance, partitioning, and conversion of nitrate across the Day River Basin (DRB), Vietnam, which is heavily impacted by agriculture and urbanization. Stable isotope compositions of river water δ¹⁸O-H₂O, in addition to their δ¹⁵N-NO₃ and δ¹⁸O-NO₃ signatures, were sampled at 12 locations in the DRB. Sample collection was conducted during three different periods to capture changes in regional weather and agricultural fertilization regimes; April (the dry season and key fertilization period), July (the rainy season and another key fertilization period) and October (the rainy season with no regional fertilization). Ranges of NO₃ stable isotopes are − 7.1 to + 9.2‰ and − 3.9 to + 13.2‰ for δ¹⁸O and δ¹⁵N, respectively. Interpretation of the stable isotope data characterizes 4 main sources of NO₃ in the DRB; (1) nitrified urea fertilizer derived from an intensive agricultural irrigation network, (2) soil and groundwater leaching from within the basin (3) manure and sewage inputs (which is more prevalent in downstream river sections) and (4) upstream inflow from the Red River which discharges into the Day River through the Dao River. We applied a mixing model for the DRB consisting of 4 variables, representing these 4 different sources. The partition calculation shows that during the fertilization and rainy period of July, more than 45% of river NO₃ is derived from nitrified urea sources. During the other sampling periods (April and October), manure and sewage contribute more than 50% of river NO₃ and are derived from the middle portion of the DRB, where the Day River receives domestic wastewater from the Vietnamese capital, Hanoi. Stable isotope data of O and N reveal that nitrification processes are more prevalent in the rainy season than in dry season and that this predominantly takes place in paddy field agricultural zones. In general, data demonstrate that nitrate loss in the DRB is due to denitrification which takes place in polluted stretches of the river and dominates in the dry season. This study highlights that (i) domestic waste should be treated prior to its discharge into the Day River and (ii) the need for better catchment agricultural fertilization practices as large portions of fertilizer currently discharge into the river, which greatly impacts regional water quality.

Appendix: Calculation of δ18O-NO3 based on water oxygen and dissolved oxygen isotopes

Nitrification occurs as a two-step process whereby ammonia is first converted to nitrite and the produced nitrite is then converted to nitrate. During the bacterial nitrification process, the biogeochemical sources of oxygen atoms are dioxygen (O₂) and water (H₂O). O₂ is incorporated during the oxidation of ammonia to hydroxylamine (NH₂OH), while H₂O is incorporated during the oxidation of both hydroxylamine to nitrite and nitrite to nitrate. While the ratio of 1:2 oxygen atoms from O₂ and H₂O implied by these observations is commonly used to interpret the oxygen isotopic content of nitrate derived from bacterial nitrification (Kendall 1998; Burns and Kendall 2002; Wankel et al. 2006), the utilization of this ratio involves the assumptions that exchange and fractionation of oxygen isotopes during nitrification are minimal. Recent works (e.g. Casciotti et al. 2007; Casciotti et al. 2010; Buchwald and Casciotti, 2010) have presented oxygen isotopic exchange and fractionation during nitrification. In general, during bacterial ammonia oxidation, the produced δ¹⁸O-NO₂ is computed as:

$${\delta }^{18}{O}_{{NO}_{2}}=\left[\frac{1}{2}\left(1+{x}_{AOB}\right)\right]\left({\delta }^{18}{O}_{{H}_{2}O}\right)+\frac{1}{2}\left({\delta }^{18}{O}_{{O}_{2}}-{}_{{k,O}_{2}}-{}_{{k,H}_{2}O,1}\right)+\left({}_{eq}\right)\left({x}_{AOB}\right)$$

(1)

In which χ_AOB, ε_k,O2,ε_k,H2O,1, ε_eq, are respectively the fraction of nitrite oxygen atoms that have equilibrated with H₂O during ammonia oxidation, the kinetic isotope effect for O₂ incorporation, the kinetic isotope effect for H₂O incorporation by hydroxylamine oxidoreductase, and the equilibrium isotope effect for nitrite equilibration with H₂O.

Then, during bacterial nitrite oxidation, δ¹⁸O-NO₃ is estimated as (exchange of oxygen atoms between nitrite and water is minimal; Buchwald & Casciotti, 2010):

$${\delta }^{18}{O}_{{NO}_{3}}=\frac{2}{3}{\delta }^{18}{O}_{{NO}_{2}}+\frac{1}{3}\left({\delta }^{18}{O}_{{H}_{2}O}-{}_{{k,H}_{2}O,2}\right)$$

(2)

whereas ε_k,H2O,2 is the kinetic isotope effect for water incorporation by nitrite oxidoreductase.

Literature review has shown that χ_AOB, ε_k,O2+ ε_k,H2O,1, ε_eq, and ε_k,H2O,2 are respectively + 0.15 ± 0.1‰, + 26.3 ± 7.7‰, + 14‰, and + 15.5 ± 3.8‰ (Casciotti et al. 2007; Casciotti et al. 2010; Buchwald & Casciotti, 2010).