The external/internal sources and sinks of greenhouse gases (CO2, CH4, N2O) in the Pearl River Estuary and adjacent coastal waters in summer.

Estuary acts as a hotspot of greenhouse gases (GHGs, including CO2, CH4 and N2O) to the atmosphere. However, the GHGs budgets, including input/output fluxes through interfaces and biogeochemical source/sink processes in water columns, of the estuarine systems are still not well constrained due to the lacking of comprehensive observational data. Here, we presented the spatial distributions of GHGs of surface/bottom water and sediment porewater along the Pearl River Estuary (PRE) and adjacent region during summertime. The incorporation of the monitoring for the sediment-water interface (SWI) with these of the water-air interface (WAI) allows us to close the budget revealing additional information of internal consumption/production processes of the three GHGs. The oversaturated CO2 (481-7573 µatm), CH4 (289-16,990 %) and N2O (108-649 %) in surface water suggested PRE is a significant GHGs source to the atmosphere, in which CO2 is the major contributor accounting for 90 % of total global warming potential (GWP), leaving 2.8 % from CH4, and 7.2 % from N2O. Addition to the river input, the SWI releases GHGs to the overlying water with fluxes of 3.5 × 107, 10.8 × 104 and 0.7 × 104 mol d-1 for CO2, CH4 and N2O, respectively. Although all three GHGs exhibited emission to the atmosphere, our mass balance calculation showed that 16.9× 107 mol d-1 of CO2 and 1.0 × 104 mol d-1 of N2O were consumed, respectively, inside the estuary water body, while extra-production (13.8 × 104 mol d-1) of CH4 was demanded in the water body to support its output flux. This is the first experiment quantitatively assessing the importance of internal carbon and nitrogen biogeochemical processes in the PRE. Our finding is of guiding significance to constrain the GHGs budget and draw up realistic pathways for modeling works of GHGs prediction.

Salinity change induces distinct climate feedbacks of nitrogen removal in saline lakes.

Current estimations of nitrogen biogeochemical cycling and N2O emissions in global lakes as well as predictions of their future changes are overrepresented by freshwater datasets, while less consideration is given to widespread saline lakes with different salinity (representing salinization or desalinization). Here, we show that N2O production by denitrification is the main process of reactive nitrogen (Nr, the general abbreviations of NH4+-N, NO2--N and NO3--N) removal in hypersaline lake sediments (e.g. Lake Chaka). The integration of our field measurements and literature data shows that in response to natural salinity decrease, potential Nr removal increases while N2O production decreases. Furthermore, denitrification-induced N2 production exhibits higher salinity sensitivity than denitrification-induced N2O production, suggesting that the contribution of N2O to Nr removal decreases with decreasing salinity. This field-investigation-based salinity response model of Nr removal indicates that under global climate change, saline lakes in the process of salinization or desalination may have distinct Nr removal and climate feedback effects: salinized lakes tend to generate a positive climate feedback, while desalinated lakes show a negative feedback. Therefore, salinity change should be considered as an important factor in assessing future trend of N2O emissions from lakes under climate change.

Permeability decides the effect of antibiotics on sedimentary nitrogen removal in Jiulong River Estuary.

Sedimentary denitrification takes place beneath the oxic layer at the sediment-water interface, where nitrate and antibiotics need to diffuse through the overlying water. However, the antibiotics' effect on sedimentary N removal and associated N2O production has not been adequately investigated under in situ conditions. Here, isotope pairing techniques, including slurry incubations (potential) and intact core incubations (in situ), combined with metagenomic analysis were applied to investigate the impacts of two protein-inhibiting antibiotics (oxytetracycline and thiamphenicol) on sediment nitrogen removal in a subtropical estuary. Slurry incubations showed that the two antibiotics significantly inhibited denitrification (67-98%) and anammox (49-99%), while intact core incubations presented no antibiotic effect at upstream but significant inhibition (23%-52%) at downstream. Meanwhile, N2O yields were stimulated up to 20 folds in slurry incubations yet showing insignificant response in intact cores. Such contrasting results between up- and down-stream and between slurry and intact core incubations strongly indicated that permeability, which determines diffusion of antibiotics to microbes, is the key to exert the effect of antibiotics on in situ sedimentary nitrogen removal processes regardless the existence of antibiotics resistance genes. This diffusive obstruction may mitigate the toxic effect of antibiotics on nitrogen removal related microbes in natural environments.

Particle-associated denitrification is the primary source of N2O in oxic coastal waters.

The heavily human-perturbed coastal oceans are hotspots of nitrous oxide (N2O) emission to the atmosphere. The processes underpinning the N2O flux, however, remain poorly understood, leading to large uncertainties in assessing global N2O budgets. Using a suite of nitrogen isotope labeling experiments, we show that multiple processes contribute to N2O production throughout the estuarine-coastal gradient, sustaining intensive N2O flux to the atmosphere. Unexpectedly, denitrification, rather than ammonia oxidation as previously assumed, constitutes the major source of N2O in well-oxygenated coastal waters. Size-fractionated manipulation experiments with gene analysis further reveal niche partitioning of ammonia oxidizers and denitrifiers across the particle size spectrum; denitrification dominated on large particles and ammonia oxidizers on small particles. Total N2O production rate increases with substrate and particle concentrations, suggesting a crucial interplay between nutrients and particles in controlling N2O production. The controlling factors identified here may help understand climate feedback mechanisms between human activity and coastal oceans.

Transcriptomic evidences for microbial carbon and nitrogen cycles in the deoxygenated seawaters of Bohai Sea.

Eutrophication-induced water deoxygenation occurs continually in coastal oceans, and alters community structure, metabolic processes, and the energy shunt, resulting in a major threat to the ecological environment. Seasonal deoxygenation events have occurred in the Bohai Sea (China), however, how these affect the functional activity of microorganisms remains unclear. Here, through the use of absolute quantification of 16S rRNA genes amplicon sequencing and metatranscriptomics approaches, we investigated the structure of the microbial community and the patterns of transcriptional activity in deoxygenated seawaters. The dominant phyla were Proteobacteria (average value, 1.4 × 106 copies ml-1), Cyanobacteria (3.7 × 105 copies ml-1), Bacteroidetes (2.7 × 105 copies ml-1), and the ammonia-oxidizing archaea Thaumarchaeota (1.9 × 105 copies ml-1). Among the various environmental factors, dissolved oxygen, pH and temperature displayed the most significant correlation with microbial community composition and functional activity. Metatranscriptomic data showed high transcriptional activity of Thaumarchaeota in the deoxygenated waters, with a significant increase in the expression of core genes representing ammonia oxidation, ammonia transport, and carbon fixation (3-hydroxypropionic acid/4-hydroxybutyric acid cycle) pathways. The transcripts of Cyanobacteria involved in photosynthesis and carbon fixation (Calvin-Benson-Bassham cycle) significantly decreased in low oxygen waters. Meanwhile, the transcripts for the ribulose bisphosphate carboxylase-encoding gene shifted from being assigned to photoautotrophic to chemoautotrophic organisms in surface and bottom waters, respectively. Moreover, the transcription profile indicated that heterotrophs play a critical role in transforming low-molecular-weight dissolved organic nitrogen. Elevated abundances of transcripts related to microbial antioxidant activity corresponded to an enhanced aerobic metabolism of Thaumarchaeota in the low oxygen seawater. In general, our transcriptional evidences showed a population increase of Thaumarchaeota, especially the coastal ecotype of ammonia oxidizers, in low oxygen aquatic environments, and indicated an enhanced contribution of chemolithoautotrophic carbon fixation to carbon flow.

Diel change in inorganic nitrogenous nutrient dynamics and associated oxygen stoichiometry along the Pearl River Estuary.

The reactive nitrogen (N) emitted from continents significantly perturbs the pristine N cycle around the land-ocean boundary resulting in eutrophication and hypoxia. As nutrients are transported downstream through an estuary, various types of biological processes co-occur to modulate nitrogen speciation to influence the biogeochemical habitats for downstream microorganisms. We surveyed the Pearl River Estuary to examine the N transfer dynamics among nitrogen species with considering process-specific oxygen production and consumption. By using 15N pulse-tracing techniques, we measured ammonia oxidation and uptakes of ammonium, nitrite, and nitrate simultaneously under dark and light conditions in parallel. Light strongly inhibited nitrification but enhanced N uptake, and such light effect was further considered in the calculation for nitrogen transformation rates over a diel cycle. We found both oxidation and uptake of ammonium decreased seaward as substrate decreased. The nitrifier and phytoplankton work in antiphase to draw down incoming ammonium rapidly. Contrary to ammonium uptake, uptake of nitrite and nitrate showed a seaward increasing pattern. Such an inverse spatial pattern implies a shift in N preference for phytoplankton. Such high ammonium preference inhibits nitrate/nitrite uptake allowing them to behave conservatively in the estuary and to travel farther to outer estuary. By integrating oxygen consumption and production induced by N transformation processes over the diel cycle, oxygen was produced although allochthonous ammonium input is high (∼250 µM). For most stations, ammonium was completely consumed within 2 days, some stations even less than 0.5 days, implying that although the water residence time is short (2-15 days), tremendous input of ammonium N from upstream was transformed into particulate organic or nitrate forms during traveling to modulate the biogeochemical niche, including substrate, organics and oxygen, of coastal microbes in water column and sediments.

Sedimentary processes dominate nitrous oxide production and emission in the hypoxic zone off the Changjiang River estuary.

Coastal oceans, known as the major nitrous oxide (N2O) source to the atmosphere, are increasingly subject to eutrophication and concurrent near-bottom hypoxia. The natural nitrogen cycle is likely to be altered markedly in hypoxic coastal oceans. However, the processes responsible for N2O production and emission remain elusive because of lacking field rate measurements simultaneously conducted in the water column and sediment. Here, we quantified N2O production rates using a 15N-labeled technique in the water-column and surface sediments off the Changjiang (Yangtze) River estuary, the largest hypoxic zone in the Pacific margins. Our results showed that the estuarine surface sediments were the major source for N2O production, accounting for approximately 90% of the total water-column accumulation and consequent efflux of N2O in the hypoxic zone, whereas the water-column nitrification and denitrification combined only contributed <10%. More importantly, the coupling of nitrification and denitrification at the presence of abundant supply and remineralization of labile organic matter was the main driver of the N2O release from the sediment-water interface in this region. This study highlights the dominant role of benthic processes occurring at the sediment-water interface controlling the coastal N2O budget, as the anthropogenic eutrophication and hypoxia are expanding in coastal oceans.

Quantitatively deciphering the roles of sediment nitrogen removal in environmental and climatic feedbacks in two subtropical estuaries.

Sedimentary denitrification and anaerobic ammonium oxidation (anammox) are two microbially-mediated nitrogen removal pathways with distinct climatic feedbacks. Estuaries receive large fluxes of anthropogenic nitrogen and serve as hotspots for nitrogen loss. Applying 15N isotope pairing technique and sediment intact core incubation in two subtropical estuaries, the Yangtze River Estuary (YRE) and Jiulong River Estuary (JRE), we show that denitrification predominates the sedimentary nitrogen loss with a minor contribution (8.6 ± 7.5%) from anammox. Particulate organic matter degradation sustains the sedimentary nitrogen removal linking the nitrogen transformations between water column and sediment. Our results indicate that estuarine sediments exhibit high areal nitrogen removal rate, but play a relatively weak role in eliminating the nitrogen inputted from river basin due to the limited area. The riverine excess nitrogen will eventually enter into the adjacent continental shelf and be removed via phytoplankton assimilation-sedimentation-degradation-coupled nitrification-denitrification. In addition, sedimentary denitrification causes 1.8 ± 2.2% of nitrogen flow towards nitrous oxide (N2O) production and the derived N2O release flux accounts for 59% and 65% of the daily sea-air N2O emission in the YRE and JRE, respectively. These findings contribute to a better understanding of estuarine sedimentary nitrogen removal and associated climate feedbacks, and to the parameterization of Earth system models.

Substrate regulation leads to differential responses of microbial ammonia-oxidizing communities to ocean warming.

In the context of continuously increasing anthropogenic nitrogen inputs, knowledge of how ammonia oxidation (AO) in the ocean responds to warming is crucial to predicting future changes in marine nitrogen biogeochemistry. Here, we show divergent thermal response patterns for marine AO across a wide onshore/offshore trophic gradient. We find ammonia oxidizer community and ambient substrate co-regulate optimum temperatures (Topt), generating distinct thermal response patterns with Topt varying from ≤14 °C to ≥34 °C. Substrate addition elevates Topt when ambient substrate is unsaturated. The thermal sensitivity of kinetic parameters allows us to predict responses of both AO rate and Topt at varying substrate and temperature below the critical temperature. A warming ocean promotes nearshore AO, while suppressing offshore AO. Our findings reconcile field inconsistencies of temperature effects on AO, suggesting that predictive biogeochemical models need to include such differential warming mechanisms on this key nitrogen cycle process.

Organic matter decomposition sustains sedimentary nitrogen loss in the Pearl River Estuary, China.

The Pearl River Estuary (PRE) has long received tremendous amounts of anthropogenic nitrogen, and is facing severe environmental problems. Denitrification and anaerobic ammonium oxidation (anammox) are known to be two major nitrogen removal pathways in estuarine sediments. Through the use of slurry and intact sediment core incubations, we examined the nitrogen removal pathways and quantified the in situ denitrification and anammox with associated gaseous nitrogen production rates. Sedimentary nitrogen removal was predominated by denitrification (93-100%) relative to a minimal contribution (<7%) from anammox. Among the detected environmental factors, salinity, bottom water NOx- (nitrate and nitrite) concentration, sedimentary organic matter and dissolved oxygen consumption rates showed good correlations with denitrification and anammox rates. Sedimentary nitrogen loss was mainly supported by endogenic coupled nitrification-denitrification (6.0 ±â€¯1.5 × 106 mol N d-1), with water-column-delivered NOx- (2.1 ±â€¯0.6 × 106 mol N d-1) as the secondary source. Such results suggested that sedimentary nitrogen removal involved mainly particulate organic form (allochthonous or autochthonous) deposited onto sediments, rather than inorganic forms in overlying water. Meanwhile, total N2O production from sediments was estimated to be 7.3 ±â€¯2.1 × 104 mol N d-1, equivalent to ~35% of the daily N2O emissions in the PRE.

Nitrogen transformations and removal efficiency enhancement of a constructed wetland in subtropical Taiwan.

Urbanization condenses reactive nitrogen into cities leaving threats of nitrogen pollution onto nearby environments when sewage is not properly treated. Constructed wetland is an ecological and economical way to remove reactive nitrogen. We investigated the seasonal nitrogen transformations and removal pathways in a surface-flow constructed wetland (93,000m2 with five treatment cells), which treats domestic wastewater in subtropical Taiwan. By using isotopic pairing technique, we found denitrification exceeds anammox dominated the nitrogen removal pathway throughout seasons. The potential denitrification (0.09 to 2.84gNm-2d-1) in the overlying water was in the same magnitude relative to that in sediments (1.26 to 4.14gNm-2d-1). The denitrification rates in sediments were highest in summer followed by autumn and winter. The concentration removal efficiencies of ammonium and dissolved inorganic nitrogen (DIN) were both highest in summer, then decreased significantly in autumn and winter. Temperature is a significant regulator for seasonal nitrogen removal. However, a positive correlation was observed between the potential denitrification rates and the amount of 15NO3- addition, indicating that nitrate addition may still stimulate denitrification under low temperature condition in winter (15.2-16.3°C). Since nitrate concentrations in porewater were much lower than that in water column for autumn and winter, we speculated NOx--N (nitrite and nitrate) supply to the sediments was a limiting factor for low DIN removal efficiency. We proposed to enhance nitrate removal efficiency via denitrification by physically promoting NOx--N and oxygen exchanges through the sediment-water interface.