Nitrate-dependent anaerobic methane oxidation and chemolithotrophic denitrification in a temperate eutrophic lake

While the emissions of methane (CH4) by the natural systems have been widely investigated, the aquatic sinks are still poorly constrained. Here, we investigated CH4 cycle and its interactions with nitrogen (N), iron (Fe) and manganese (Mn) cycles in the oxic-anoxic interface and deep anoxic waters of a small, meromictic and eutrophic lake, during two summertime sampling campaigns. Anaerobic CH4 oxidation (AOM) was measured from the temporal decrease of CH4 concentrations, with addition of 3 potential electron acceptors (NO3–, iron oxides (Fe(OH)3) and manganese oxides (MnO2)). Experiments with addition of either 15N-labeled nitrate (15N-NO3–) or 15N-NO3– combined with sulfide (H2S), to measure denitrification, chemolithotrophic denitrification and anaerobic ammonium oxidation (anammox) rates were also performed. Measurements showed AOM rates up to 3.8 µmol CH4 L–1 d–1 that strongly increased with the addition of NO3– and moderately increased with the addition of Fe(OH)3. No stimulation was observed with MnO2 added. Potential denitrification and anammox rates up to 63 and 0.27 µmol N2 L–1 d–1, respectively, were measured when only 15N-NO3– was added. When H2S was added, both denitrification and anammox rates increased. Altogether, these results suggest that prokaryote communities in the redoxcline are able to efficiently use the most available substrates.

No-Till and Solid Digestate Amendment Selectively Affect the Potential Denitrification Activity in Two Mediterranean Orchard Soils

Improved soil managements that include reduced soil disturbance and organic amendment incorporation represent valuable strategies to counteract soil degradation processes that affect Mediterranean tree cultivations. However, changes induced by these practices can promote soil N loss through denitrification. Our research aimed to investigate the short-term effects of no-tillage and organic amendment with solid anaerobic digestate on the potential denitrification in two Mediterranean orchard soils showing contrasting properties in terms of texture and pH. Denitrifying enzyme activity (DEA) and selected soil variables (available C and N, microbial biomass C, basal respiration) were monitored in olive and orange tree orchard soils over a five-month period. Our results showed that the application of both practices increased soil DEA, with dynamics that varied according to the soil type. Increased bulk density, lowered soil aeration, and a promoting effect on soil microbial community growth were the main DEA triggers under no-tillage. Conversely, addition of digestate promoted DEA by increasing readily available C and N with a shorter effect in the olive grove soil, due to greater sorption and higher microbial efficiency, and a long-lasting consequence in the orange orchard soil related to a larger release of soluble substrates and their lower microbial use efficiency.

Contribution of Particulate and Mineral-Associated Organic Matter to Potential Denitrification of Agricultural Soils

Water-extractable organic carbon (WEOC) is considered as the most important carbon (C) source for denitrifying organisms, but the contribution of individual organic matter (OM) fractions (i.e., particulate (POM) and mineral-associated (MOM)) to its release and, thus, to denitrification remains unresolved. Here we tested short-time effects of POM and MOM on potential denitrification and estimated the contribution of POM- and MOM-derived WEOC to denitrification and CO2 production of three agricultural topsoils. Suspensions of bulk soils with and without addition of soil-derived POM or MOM were incubated for 24 h under anoxic conditions. Acetylene inhibition was used to determine the potential denitrification and respective product ratio at constant nitrate supply. Normalized to added OC, effects of POM on CO2 production, total denitrification, and its product ratios were much stronger than those of MOM. While the addition of OM generally increased the (N2O + N2)-N/CO2-C ratio, the N2O/(N2O + N2) ratio changed differently depending on the soil. Gas emissions and the respective shares of initial WEOC were then used to estimate the contribution of POM and MOM-derived WEOC to total CO2, N2O, and N2O + N2 production. Water-extractable OC derived from POM accounted for 53–85% of total denitrification and WEOC released from MOM accounted for 15–47%. Total gas emissions from bulk soils were partly over- or underestimated, mainly due to nonproportional responses of denitrification to the addition of individual OM fractions. Our findings show that MOM plays a role in providing organic substrates during denitrification but is generally less dominant than POM. We conclude that the denitrification potential of soils is not predictable based on the C distribution over POM and MOM alone. Instead, the source strength of POM and MOM for WEOC plus the WEOC’s quality turned out as the most decisive determinants of potential denitrification.

Copper nanoparticles prompt the activity of estuarine denitrifying bacterial communities at relevant environmental concentrations

Effects of metallic nanoparticles (NPs) to the estuarine biota have mostly been shown for concentrations higher than those actually measured or predicted in these environments. To address this gap, a range of concentrations expected to occur in estuarine environments (from 0.01 to 1 μg g-1) was employed in microcosms studies to assess the impact of Cu NPs in the denitrification pathway. That was achieved by quantifying gene expression and the potential denitrification rate in estuarine sediments exposed to Cu NPs for up to six days. Expression of nitrite (nirS) and nitrous oxide (nosZ) reductase genes was enhanced in a timewise manner. For the highest Cu NPs (1 μg g-1) an increase in gene expression could be seen immediately after 1 h of exposure, and continuing to be enhanced up until 7 h of exposure. For the lowest Cu NPs (0.01 μg g-1) an increase in gene expression could only be seen after 4 h or 7 h of exposure; however it continued to rise up until 24 h of exposure. In any case, after 48 h the expression levels were no longer different from the non-exposed control. Concomitantly to increased gene expression the potential denitrification rate was increased by 30 %. Our results suggest that deposition and adsorption of Cu NPs to estuarine sediments promotes the immediate and transient expression of key genes of the denitrification pathway. The long term impact of continuous inputs of Cu NPs into estuaries deserves renewed analysis to account for their effects, not just on the biota, but especially on ecosystems services.Environmental significanceInteractions of metallic nanoparticles with microbial communities of estuarine sediments are poorly characterized and its impact towards ecosystem services even less. By assessing the effect of copper nanoparticles on the expression of key genes of the denitrification pathway, an essential step for nitrogen (N) removal, we were able to show that denitrifying communities are immediately activated after exposure, increasing the denitrification rates in estuaries. The importance of denitrification lies in its release of dinitrogen (N2) to the atmosphere but also in the emissions of N2O (a potent greenhouse gas). The results obtained in this study gather data that contribute information on the denitrification dynamics in estuaries, invaluable for a timely response to the expected upcoming changes in coastal areas.Table of contentsIn estuaries the deposition upon the sediments of copper nanoparticles can contribute to change metal availability and promote the activity of denitrifying bacteria

Denitrification induced by plant residues is driven by water-soluble organic carbon

<p>Denitrification usually takes place under anoxic conditions and over short periods of time and depends on readily available nitrate and carbon sources. Variations in CO<sub>2</sub> and N<sub>2</sub>O emissions from soils amended with plant residues have mainly been explained by differences in their decomposability. Another factor rarely considered so far is water-extractable organic matter (WEOM) released into soil during residue decomposition. Here, we examined the potential effect of plant residues on denitrification with special emphasis on WEOM. A range of fresh and leached plant residues was characterized by elemental analyses, <sup>13</sup>C-NMR spectroscopy, and extraction with ultrapure water. The obtained solutions were analyzed for the concentration of organic carbon (OC), organic nitrogen (ON), and by UV-VIS spectroscopy. To test the potential denitrification induced by plant residues or three different OM solutions, these carbon sources were added to soil suspensions and incubated for 24 hours at 20 °C in the dark under anoxic conditions; KNO<sub>3</sub> was added to ensure unlimited nitrate supply. Evolving N<sub>2</sub>O and CO<sub>2</sub> were analyzed by gas chromatography and acetylene inhibition was used to determine denitrification and its product ratio. The production of all gases as well as the molar N<sub>2</sub>O+N<sub>2</sub>-N/CO<sub>2</sub>-C ratio was directly related to the water-extractable OC (WEOC) content of the plant residues and the WEOC increased with carboxylic/carbonyl C and decreasing OC/ON ratios of the plant residues. Incubation of OM solutions revealed that the molar N<sub>2</sub>O+N<sub>2</sub>-N/CO<sub>2</sub>-C ratio and share of N<sub>2</sub>O are influenced by the WEOM’s chemical composition. In conclusion, the effect of plant residues on potential denitrification is governed by their composition and the related production of WEOM.</p>

Seasonal comparison of potential denitrification rates and nitrogen functional genes with sediment depths in the wetland, Korea

<p>Wetlands provide not only habitats for a wide range of organisms but also ecological functions of degrading and removing pollutants from water body through a variety of physical, chemical and biological processes. Seasonal variation including recent increases in the frequency of floods and droughts have affected the hydrological environment of wetlands. Furthermore, these effects may result in changes in redox conditions in the nitrogen biogeochemical process in wetland sediments. Therefore, in this study, the potential denitrification rate and denitrification-related gene quantitative analysis was performed to investigate seasonal nitrogen dynamics of wetland sediments associated with surface and groundwater interactions in Baekseok reservoir wetlands (Gunsan-si, Jeollabuk Province, Korea). Sediment from two different sites (i.e., PA and PB) in wetland were collected with different depths in June and December 2019 to investigate seasonal effects on denitrification with sediment depths. Potential denitrification rate experiments were performed using the acetylene inhibition technique, and denitrification-related gene quantification was performed by qPCR analysis. As a result of potential denitrification rate, PA sites ranged from 2.67–3.27 ng N<sub>2</sub>O/g/hr and 3.13–15.13 ng N<sub>2</sub>O/g/hr in June and December, respectively. PB sites ranged from 2.43-6.30 ng N<sub>2</sub>O/g/hr and 5.47-6.30 ng N<sub>2</sub>O/g/hr. Overall, higher levels were observed at 0–10 cm, with higher denitrification rates in December than in June. The qPCR analysis showed that the narG, nirS and nosZ gene copy number ranges for the PA site in June showed 1.82 x 10<sup>6</sup> – 6.15 x 10<sup>7</sup> copies/g, and in December 7.71 x 10<sup>5</sup> – 5.97 x 10<sup>8</sup> copies/g. The narG, nirS and nosZ gene copy number ranges for the PB site in June showed 3.53 x 10<sup>5</sup> – 3.86 x 10<sup>8</sup> copies/g, and in December, 1.24 x 10<sup>6</sup> – 3.47 x 10<sup>8</sup> copies/g. Overall, both sites had higher copy numbers in December than in June, corresponding to an increase in potential denitrification rate in December.</p>