Xiaofen Jiang

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.

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.

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.

Effects of thiamphenicol on nitrate reduction and N2O release in estuarine and coastal sediments.

Nitrate overload is an important driver of water pollution in most estuarine and coastal ecosystems, and thus nitrate reduction processes have attracted considerable attention. Antibiotics contamination is also an emerging environmental problem in estuarine and coastal regions as a result of growing production and usage of antibiotics. However, the effects of antibiotics on nitrate reduction remain unclear in these aquatic ecosystems. In this study, continuous-flow experiments were conducted to examine the effects of thiamphenicol (TAP, a common chloramphenicol antibiotic) on nitrate reduction and greenhouse gas N2O release. Functional genes involved in nitrogen transformation were also quantified to explore the microbial mechanisms of the TAP influence. Production of N2 were observed to be inhibited by TAP treatment, which implied the inhibition effect of TAP on nitrate reduction processes. As intermediate products of nitrogen transformation processes, nitrite and N2O were observed to accumulate during the incubation. Different TAP inhibition effects on related functional genes may be the microbial mechanism for the changes of nutrient fluxes, N2 fluxes and N2O release rates. These results indicate that the antibiotics residues in estuarine and coastal ecosystems may contribute to nitrate retention and N2O release, which could be a major factor responsible for eutrophication and greenhouse effects.

Effects of multiple antibiotics exposure on denitrification process in the Yangtze Estuary sediments

Denitrification is a dominant reactive nitrogen removal pathway in most estuarine and coastal ecosystems, and plays a significant role in regulating N2O release. Although multiple antibiotics residues are widely detected in aquatic environment, combined effects of antibiotics on denitrification remain indistinct. In this work, 5 classes of antibiotics (sulfonamides, chloramphenicols, tetracyclines, macrolides, and fluoroquinolones) were selected to conduct orthogonal experiments in order to explore their combined effects on denitrification. 15N-based denitrification and N2O release rates were determined in the orthogonal experiments, while denitrifying functional genes were examined to illustrate the microbial mechanism of the combined antibiotics effect. Denitrification rates were inhibited by antibiotics treatments, and synergistic inhibition effect was observed for multiple antibiotics exposure. Different classes of antibiotics had different influence on N2O release rates, but multiple antibiotics exposure mostly led to stimulatory effect. Abundances of denitrifying functional genes were inhibited by multiple antibiotics exposure due to the antimicrobial properties, and different inhibition on denitrifiers may be the major mechanism for the variations of N2O release rates. Combined effects of antibiotics on denitrification may lead to nitrate retention and N2O release in estuarine and coastal ecosystems, and consequently cause cascading environmental problems, such as greenhouse effects and hyper-eutrophication.

DNRA in intertidal sediments of the Yangtze Estuary

Dissimilatory nitrate reduction to ammonium (DNRA) plays an important role in regulating the fate of reactive nitrogen in estuarine and coastal ecosystems. In this work, intertidal sediments of the Yangtze Estuary were collected in January and August of 2015, respectively. Potential rates of DNRA and associated functional gene were investigated with nitrogen isotope-tracing and molecular techniques. The measured DNRA rates ranged from 0.14 to 5.57 μmol 15N kg-1 h-1 in the intertidal sediments. DNRA rates were tightly related to abundance of nrfA gene (p <0.001), demonstrating that fermentation reaction may be the dominant pathway of DNRA in the study area. Redundancy analysis (RDA) showed a relationship between DNRA and organic matter and NO2-, suggesting that these substrates stimulated the metabolism of DNRA microorganisms. On the other hand, the correlation between abundances of nrfA gene and Fe2+ and sulfide in the RDA analysis implied that oxidation of both Fe2+ and sulfide can enhance fermentative DNRA by providing extra free energy. DNRA converted approximately 2.29 × 105 t of nitrate to ammonia annually in the sampling area of the Yangtze Estuary, and most of the converted ammonium was retained in the estuarine ecosystem. DNRA may further contribute to eutrophication in the Yangtze Estuary, and also in the other hypereutrophic estuaries.