Xianbiao Lin

Dissimilatory nitrate reduction processes and associated contribution to nitrogen removal in sediments of the Yangtze Estuary

Dissimilatory nitrate reduction processes, including denitrification, anaerobic ammonium oxidation (ANAMMOX), and dissimilatory nitrate reduction to ammonium (DNRA), play an important role in controlling the nitrate dynamics and fate in estuarine and coastal environments. We investigated potential rates of denitrification, ANAMMOX, and DNRA in the sediments of the Yangtze Estuary via slurry incubation experiments combined with isotope-tracing techniques to reveal their respective contributions to total nitrate reduction in this hypereutrophic estuarine ecosystem. Measured rates of denitrification, ANAMMOX, and DNRA ranged from 0.06 to 4.51 µmol N kg−1 h−1, 0.01 to 0.52 µmol N kg−1 h−1, and 0.03 to 0.89 µmol N kg−1 h−1, respectively. These potential dissimilatory nitrate reduction process rates correlated significantly with salinity, sulfide, organic carbon, and nitrogen. Denitrification contributed 38–96% total nitrate reduction in the Yangtze Estuary, as compared to 3–45% for DNRA and 1–36% for ANAMMOX. In total, the denitrification and ANAMMOX processes removed approximately 25% of the external inorganic nitrogen transported annually into the estuary. In contrast, most external inorganic nitrogen was retained in the estuary and contributes substantially to the severe eutrophication of the Yangtze Estuary.

Anaerobic ammonium oxidation and its contribution to nitrogen removal in China's coastal wetlands.

Over the past several decades, human activities have caused substantial enrichment of reactive nitrogen in China’s coastal wetlands. Although anaerobic ammonium oxidation (anammox), the process of oxidizing ammonium into dinitrogen gas through the reduction of nitrite, is identified as an important process for removing reactive nitrogen, little is known about the dynamics of anammox and its contribution to nitrogen removal in nitrogen-enriched environments. Here, we examine potential rates of anammox and associate them with bacterial diversity and abundance across the coastal wetlands of China using molecular and isotope tracing techniques. High anammox bacterial diversity was detected in China’s coastal wetlands and included Candidatus Scalindua, Kuenenia, Brocadia, and Jettenia. Potential anammox rates were more closely associated with the abundance of anammox bacteria than to their diversity. Among all measured environmental variables, temperature was a key environmental factor, causing a latitudinal distribution of the anammox bacterial community composition, biodiversity and activity along the coastal wetlands of China. Based on nitrogen isotope tracing experiments, anammox was estimated to account for approximately 3.8–10.7% of the total reactive nitrogen removal in the study area. Combined with denitrification, anammox can remove 20.7% of the total external terrigenous inorganic nitrogen annually transported into China’s coastal wetland ecosystems.

Evidence of Nitrogen Loss from Anaerobic Ammonium Oxidation Coupled with Ferric Iron Reduction in an Intertidal Wetland

Anaerobic ammonium oxidation coupled with nitrite reduction is an important microbial pathway of nitrogen removal in intertidal wetlands. However, little is known about the role of anaerobic ammonium oxidation coupled with ferric iron reduction (termed Feammox) in intertidal nitrogen cycling. In this study, sediment slurry incubation experiments were combined with an isotope-tracing technique to examine the dynamics of Feammox and its association with tidal fluctuations in the intertidal wetland of the Yangtze Estuary. Feammox was detected in the intertidal wetland sediments, with potential rates of 0.24-0.36 mg N kg(-1) d(-1). The Feammox rates in the sediments were generally higher during spring tides than during neap tides. The tidal fluctuations affected the growth of iron-reducing bacteria and reduction of ferric iron, which mediated Feammox activity and the associated nitrogen loss from intertidal wetlands to the atmosphere. An estimated loss of 11.5-18 t N km(-2) year(-1) was linked to Feammox, accounting for approximately 3.1-4.9% of the total external inorganic nitrogen transported into the Yangtze Estuary wetland each year. Overall, the co-occurrence of ferric iron reduction and ammonium oxidation suggests that Feammox can act as an ammonium removal mechanism in intertidal wetlands.

Primary effects of extracellular enzyme activity and microbial community on carbon and nitrogen mineralization in estuarine and tidal wetlands

Estuarine and tidal wetlands with high primary productivity and biological activity play a crucial role in coastal nutrient dynamics. Here, to better reveal the effects of extracellular enzymes and microbial community on carbon (C) and nitrogen (N) mineralization, the incubation experiments with different C and N addition patterns to the tidal sediments of the Yangtze Estuary (China) were conducted. The results suggested a significant increase in cumulative CO2 effluxes in the C and CN treatment experiments, while no significant difference in cumulative CO2 effluxes between the N treatment and control (CK) experiments was observed. In addition, the nutrient addition patterns had a great influence on dissolve organic C and N levels, but a small effect on microbial biomass C and N. Microbial community composition and microbial activity were found to be positively correlated with organic C (OC) and the molar ratio of C to N (C/N). Partial correlation analysis, controlling for C/N, supported direct effects of OC on the activity of carbon-cycling extracellular enzymes (cellulase and polyphenol oxidase), while C/N exhibited negatively correlations with urease and Gram-positive bacteria to Gram-negative bacteria (G+/G−). Strong relationships were found between CO2 efflux and mineral nitrogen with the activity of specific enzymes (sucrase, cellulase, and polyphenol oxidase) and abundances of Gram-negative bacteria, arbuscular mycorrhizal fungi, and fungi, suggesting the significant influences of microbial community and enzyme activity on C and N mineralization in the estuarine and tidal wetlands. Furthermore, this study could highlight the need to explore effects of nutrient supply on microbial communities and enzyme activity changes associated with the C and N mineralization in these wetlands induced by the climate change.

Occurrence of matrix-bound phosphine in intertidal sediments of the Yangtze Estuary.

This study investigated the levels and potential transformation of matrix-bound phosphine in the intertidal sediments (0-5cm) of the Yangtze Estuary. Matrix-bound phosphine concentrations in sediments ranged from 0.65 to 3.25ngkg(-1), with an annual average of 1.53ngkg(-1). In freshwater sediments, the concentrations of matrix-bound phosphine were significantly higher than in the brackish sediments. The maximum concentrations of matrix-bound phosphine appeared in July (1.17-3.25ngkg(-1)), followed by May (0.92-3.01ngkg(-1)), November (0.65-2.41ngkg(-1)) and January (0.51-1.42ngkg(-1)). Matrix-bound phosphine derived probably from the mechanochemical reduction of apatite-bound phosphate and the microbial conversion of organic phosphorus in the intertidal sediments. Its spatial and seasonal distributions, however, were regulated by salinity and sediment temperature. Compared with other aquatic systems (e.g. rivers, lakes and coastal seas), a low level of matrix-bound phosphine was observed in the intertidal sediments, probably implicating a relatively rapid turnover of phosphine in the system.

Tidal pumping facilitates dissimilatory nitrate reduction in intertidal marshes

Intertidal marshes are alternately exposed and submerged due to periodic ebb and flood tides. The tidal cycle is important in controlling the biogeochemical processes of these ecosystems. Intertidal sediments are important hotspots of dissimilatory nitrate reduction and interacting nitrogen cycling microorganisms, but the effect of tides on dissimilatory nitrate reduction, including denitrification, anaerobic ammonium oxidation and dissimilatory nitrate reduction to ammonium, remains unexplored in these habitats. Here, we use isotope-tracing and molecular approaches simultaneously to show that both nitrate-reduction activities and associated functional bacterial abundances are enhanced at the sediment-tidal water interface and at the tide-induced groundwater fluctuating layer. This pattern suggests that tidal pumping may sustain dissimilatory nitrate reduction in intertidal zones. The tidal effect is supported further by nutrient profiles, fluctuations in nitrogen components over flood-ebb tidal cycles, and tidal simulation experiments. This study demonstrates the importance of tides in regulating the dynamics of dissimilatory nitrate-reducing pathways and thus provides new insights into the biogeochemical cycles of nitrogen and other elements in intertidal marshes.

Diversity, Abundance, and Distribution of nirS-Harboring Denitrifiers in Intertidal Sediments of the Yangtze Estuary

Denitrification plays a critical role in nitrogen removal in estuarine and coastal ecosystems. In this study, the community composition, diversity, abundance, and distribution of cytochrome cd1-type nitrite reductase gene (nirS)-harboring denitrifiers in intertidal sediments of the Yangtze Estuary were analyzed using polymerase chain reaction (PCR)-based clone libraries and quantitative PCR techniques. Clone library analysis showed that the nirS-encoding bacterial biodiversity was significantly higher at the lower salinity sites than at the higher salinity sites. However, there was no significant seasonal difference in the nirS gene diversity between summer and winter. Phylogenetic analysis revealed that the nirS-harboring denitrifier communities at the study area had distinctive spatial heterogeneity along the estuary. At the lower salinity sites, the nirS-harboring bacterial community was co-dominated by clusters III and VII; while at the higher salinity sites, it was dominated by cluster I. Canonical correspondence analysis indicated that the community compositions of nirS-type denitrifiers were significantly correlated with salinity, ammonium, and nitrate. Quantitative PCR results showed that the nirS gene abundance was in the range of 1.01 × 106 to 9.00 × 107 copies per gram dry sediment, without significant seasonal variation. Among all the environmental factors, the nirS gene abundance was only significantly related to the change of salinity. These results can extend our current knowledge about the composition and dynamics of denitrification microbial community in the estuarine ecosystem.

Polycyclic aromatic hydrocarbons and black carbon in intertidal sediments of China coastal zones: Concentration, ecological risk, source and their relationship

Polycyclic aromatic hydrocarbons (PAHs) and black carbon (BC) have attracted many attentions, especially in the coastal environments. In this study, spatiotemporal distributions of PAHs and BC, and the correlations between BC and PAHs were investigated in the intertidal sediments of China coastal zones. BC in sediments was measured through dichromate oxidation (BCCr) and thermal oxidation (BCCTO). The concentrations of BCCr in the intertidal sediments ranged between 0.61 and 6.32mgg(-1), while BCCTO ranged between 0.57 and 4.76mgg(-1). Spatial variations of δ(13)C signatures in TOC and BC were observed, varying from -21.13‰ to -24.87‰ and from -23.53‰ to -16.78‰, respectively. PAH contents of sediments ranged from 195.9 to 4610.2ngg(-1) in winter and 98.2 to 2796.5ngg(-1) in summer, and significantly seasonal variations were observed at most sampling sites. However, the results of potential toxicity assessment indicated low ecological risk in the intertidal sediments of China coastal zones. Greater concentrations of PAHs measured in the sediments of estuarine environments indicated that rivers runoff may have been responsible for the higher PAH pollution levels in the intertidal sediments of China coastal zones. Pearsons correlation analysis suggested that pyrogenic compounds of PAH were significantly related to BC, due to that both BC and these compounds derived mainly from the combustion process of fossil fuels and biomass. Overall, increasing energy consumptions caused by anthropogenic activities can contribute more emissions of BC as well as PAHs and thus improve the importance of BC in indicating pyrogenic compounds of PAHs in the intertidal sediments of China coastal zones.

Net anthropogenic nitrogen inputs (NANI) into the Yangtze River basin and the relationship with riverine nitrogen export

This study investigated net anthropogenic nitrogen inputs (NANI, including atmospheric nitrogen deposition, nitrogenous fertilizer use, net nitrogen import in food and feed, and agricultural nitrogen fixation) and the associated relationship with riverine dissolved inorganic nitrogen (DIN) export in the Yangtze River basin during the 1980–2012 period. The total NANI in the Yangtze River basin has increased by more than twofold over the past three decades (3537.0 ± 615.3 to 8176.6 ± 1442.1 kg N km−2 yr−1). The application of chemical fertilizer was the largest component of NANI in the basin (51.1%), followed by net nitrogen import in food and feed (26.0%), atmospheric nitrogen deposition (13.2%), and agricultural nitrogen fixation (9.7%). A regression analysis showed that the riverine DIN export was strongly correlated with NANI and the annual water discharge (R2 = 0.90, p < 0.01). NANI in the Yangtze River basin was estimated to contribute 37–66% to the riverine DIN export. We also forecasted future variations in NANI and riverine DIN export for the years 2013 to 2030, based on possible future changes in human activities and the climate. This work provides a quantitative understanding of NANI in the Yangtze River basin and its effects on riverine DIN export and helps to develop integrated watershed nitrogen management strategies.

Nitrogen mineralization and immobilization in sediments of the East China Sea: Spatiotemporal variations and environmental implications

Nitrogen (N) mineralization and immobilization are important processes of N biogeochemical cycle in marine sediments. This study investigated gross N mineralization (GNM) and NH4+ immobilization (GAI) in the sediments from the East China Sea (ESC), using 15N stable isotope dilution technique. Results show that measured rates of GNM and GAI ranged from 0.04 to 6.1 µg N g−1 d−1 and from undetectable to 9.82 µg N g−1 d−1, respectively. In general, both GNM and GAI rates were significantly greater in summer as compared to winter, and the high rates occurred mainly in the muddy area and increased gradually from the Yangtze Estuary to Zhe-Min Coastal muddy areas. The GNM and GAI processes were related closely to sediment temperature, pH, ammonium (NH4+), nitrate (NO3−), total organic carbon (TOC), and total nitrogen (TN) contents in the muddy area, while they were associated tightly with sediment temperature, pH, NH4+, TOC, TN, sulfide, and Fe(III) concentrations in the sandy area. In addition, the total mineralized and immobilized N in the East China Sea (ECS) were estimated to be approximately 2.1 × 106 t N yr−1 and 2.7 × 106 t N yr−1, respectively. Overall, these results highlight the importance of N mineralization and immobilization in controlling the N budget in the ECS and improve the understanding of both processes and associated controlling mechanisms in the coastal marine ecosystem.