Graphene oxide stimulated low-temperature denitrification activity of microbial communities in lake sediments by enhancing anabolism and inhibiting cellular respiration.

Nitrate pollution in fresh water is becoming increasingly serious. In this study, the effects of temperature and graphene oxide materials on the potential functions of denitrification communities in lake sediments were investigated by metagenome. The addition of graphene oxide significantly affected the abundance of denitrification genes such as Nap, Nos, and enhanced the contribution of Pseudomonas, making low temperature and material addition conducive to the denitrification process. Module network implied that low temperature increased the centrality of denitrification in community functions. At low temperatures, graphene oxide enhanced community anabolism by stimulation organic carbon consumption and regulating the gene abundance in the citric acid cycle and the semi-phosphorylation Entner-Doudoroff, thus possibly stimulating extracellular polymeric substances (EPS) synthesis and secretion. In addition, graphene oxide may also regulate the transfer of reducing electrons from NADH to denitrifying enzymes by affecting the gene abundances of complex I and complex IV.

Performance, microbial growth and community interactions of iron-dependent denitrification in freshwaters.

Iron-dependent denitrification is a safe and promising technology for nitrogen removal in freshwaters. However, the understanding of microbial physiology and interactions during the process was limited. Denitrifying systems inoculated with freshwater samples were operated with and without iron(II) at a low C/N ratio for 54 days. Iron addition improved nitrogen removal. Batch experiments confirmed that microbially mediated reaction rather than abiotic reaction dominated during the process. Metagenomics recovered genomes of the five most abundant microorganisms, which accounted for over 99% of the community in every triplicate of the iron-based system. Based on codon usage bias, all of them were fast-growing organisms. The total abundance of fast-growing organisms was 38% higher in the system with iron than in the system without iron. Notably, the most abundant organism Diaphorobacter did not have enzymes for asparagine and aspartate biosynthesis, whereas Rhodanobacter could not produce serine and cobalamin. Algoriphagus and Areminomonas lost synthesis enzymes for more amino acids and vitamins. However, they could always obtain these growth-required substances from another microorganism in the community. The two-partner relationship minimized the limitation on microbial reproduction and increased community stability. Our results indicated that iron addition improved nitrogen removal by supplying electron donors, promoting microbial growth, and building up syntrophic interactions among microorganisms with timely communications. The findings provided new insights into the process, with implications for freshwater remediation.

Life strategies and metabolic interactions of core microbes during thiosulphate-based denitrification.

Sulphur-driven denitrification is a low-cost process for the treatment of nitrate-contaminated water. However, a comprehensive understanding of core populations and microbial interactions of a sulphur-based denitrifying system is lacking. This study presents results from three replicated denitrifying systems amended with thiosulphate and operated under a low C/N ratio. Amplicon sequencing revealed gradual enrichments of a few abundant denitrifiers. Based on genome-centred metagenomics and metatranscriptomics, a core set of microbes was identified in the systems, with Pseudomonas 1 and Thauera 2 being the most abundant ones. Although the replicates showed different enrichments, generalized observations were summarized. Most core populations conserved energy from denitrification coupled with sulphur. Pseudomonas 1 and Thauera 2 were able to finish complete denitrification. Surprisingly, they were also able to synthesize almost all amino acids and vitamins. In contrast, less abundant members, including Pseudomonas 2, were relatively auxotrophic and required an exogenous supply of amino acids and vitamins. The high expression of enzymes involved in biosynthesis and transport systems indicated their syntrophic relationships. The genomic findings suggested life strategies and interactions of the core thiosulphate-based denitrifying microbiome, with implications for nitrate-polluted water remediation.

Effect of low temperature on contributions of ammonia oxidizing archaea and bacteria to nitrous oxide in constructed wetlands.

Constructed wetlands (CWs) have been widely used for ecological remediation of micro-polluted source water. Nitrous oxide (N2O) from CWs has caused great concern as a greenhouse gas. However, the contribution of ammonia oxidation driven by ammonia oxidizing archaea (AOA) and ammonia oxidizing bacteria (AOB) to N2O emission, especially at low temperature, was unknown. This study aimed to quantify the contributions of AOA and AOB to N2O through lab-scale subsurface CWs. The N2O emission flux of CW at 8 °C was 1.23 mg m-2·h-1, significantly lower than that at 25 °C (1.92 mg m-2·h-1). The contribution of ammonia oxidation to N2O at 8 °C (33.04%) was significantly higher than that at 25 °C (24.17%). The N2O production from AOA increased from 1.91 ng N·g-1 at 25 °C to 4.11 ng N·g-1 soil at 8 °C and its contribution increased from 23.38% to 30.18% (P < 0.05). Low temperature impaired functional gene groups and inhibited the activity of AOB, resulting in its declined contribution. Based on the transcriptional analysis, AOA was less affected by low temperature, thus stably contributing to N2O. Moreover, community diversity and relationships of AOA were enhanced at 8 °C, while AOB declined. The results confirmed the significant contribution of AOA and demonstrated molecular mechanisms (higher activity and community stability) of the increased contribution of AOA to N2O at low temperature.

Using hierarchical stable isotope to reveal microbial food web structure and trophic transfer efficiency differences during lake melt season.

The microbial food web (MFW) is a material and energy source in lake water ecosystems. Although it is crucial to determine its structure and function for water ecological health, MFW changes during lake melt period have not been well studied. In this study, the MFW was divided into three categories by analyzing its structure and trophic transfer efficiency using hierarchical C/N stable isotopes and eDNA sequencing techniques, including the detrital food web (DFC, 15 %), classical grazing food web (CFC, 60 %), and mixed trophic food web (MFC, 25 %). The trophic structure and type of MFW in ice-melting lakes are always in the process of succession and adaptation, which is in a relatively low trophic transfer efficiency stage under stable conditions (i.e. CFC), whereas the input of exogenous debris and organic pollutants may lead to an increase in MFW trophic transfer efficiency (i.e. MFC, DFC). The trophic transfer efficiency from the previous trophic level to protozoa and micrometazoa was 16.32 % and 20.77 % in DFC and 10.20 % and 29.43 % in MFC, respectively. Both are obviously higher than those of the CFC (11.69 % and 9.45 %, respectively). In terms of trophic structure, the community interaction and trophic cascade effect of DFC and MFC were enhanced but easily changed with environmental factors. In contrast, the core species and cascading effects of the CFC were clearer, and the MFW structure was relatively stable. Overall, this study reveals that the explosive increase in MFW trophic transfer efficiency induced by exogenous input during the lake melt period may subsequently lead to the destabilization of the microbial community structure and cause potential ecological risks. These are manifested in the absence of ecological trophic processes, the decrease in trophic structure complexity and stability, and the weakening of microecology self-adaptive regulation ability.

Archaeal and bacterial ecological strategies in sediment denitrification under the influence of graphene oxide and different temperatures.

As an emerging material, graphene oxide (GO) has been widely used in recent years and will inevitably enter into natural water bodies, and it may have an impact on lake microbial communities owing to its potential toxicity and denitrification-enhancing ability. This study simulated the effect of 0.1 g/L GO on denitrification in lake sediments under summer (28 °C) and winter temperatures (8 °C). GO promoted carbon source metabolism and denitrification. Phylogenetic bin-based null model analysis suggested that GO significantly altered the contribution of heterogeneous selection in bacterial and archaeal community assembly. The co-occurrence network indicated that bacterial communities responded to the enhancement of heterogeneous selection by strategies of enhancing positive correlation and shared niche, whereas archaeal communities adopted strategies of enhancing negative correlation and competition. Bacterial networks also emerged with more non-hub connector species that could drive changes in community structure. Our study contributed to the understanding of different ecological strategies adopted by bacterial and archaeal communities in response to changes in ecological selection driven by GO.

Denitrification and dissimilatory nitrate reduction to ammonia in long-term lake sediment microcosms with iron(II).

Nitrate is an abundant pollutant in aquatic environments. Competition between the nitrate reduction processes, denitrification, which converts nitrate into nitrogen gas, and dissimilatory nitrate reduction to ammonia (DNRA), which converts nitrate into ammonia, decides whether an ecosystem removes or retains nitrogen. The presence of iron was previously reported to stimulate DNRA while sometimes inhibiting denitrification in in-situ studies, but long-term effect of iron(II) inputs on the competition is unknown. Here we inoculated long-term microcosms with sediments from two freshwater lakes. During 540 days of incubations, the microcosms with nitrate and Fe(II) additions of both lakes were able to sustain high nitrate reduction rates. Lepidocrocite was produced as a product of iron oxidation. We found both denitrification and DNRA were stimulated by nitrate and iron in the absence of external organic carbon addition. Phylogenetic analysis of denitrification genes, nirK and nirS, and DNRA genes, nirB and nrfA, was performed with metagenomic sequencing results. Enrichment was shown for reported Fe(II)-dependent nitrate reducers associated with nirS and nirB. Most of these bacteria are affiliated with Betaproteobacteria. From 16S rRNA gene analysis, Betaproteobacteria was enriched as well. In parallel, heterotrophic denitrifiers and methanotrophic DNRA archaea increased in abundance. Our results suggested heterotrophic and Fe(II)-dependent nitrate reducers both contributed to denitrification and DNRA in long-term microcosm incubations provided with iron.

The promotion and inhibition effect of graphene oxide on the process of microbial denitrification at low temperature.

This study found that graphene oxide (GO) improved microbial denitrification at low temperatures (~12 °C), and the optimal concentration was 10 mg/L as the removal rate of NO3-N increased by 17%. At the optimal concentration, GO improved the electron transport system activity of the microbes and enhanced the activity of nitrate reductase and nitrite reductase while exhibited low microbial toxicity. The addition of GO increased the content of tightly bound extracellular polymeric substances (EPS). The results of fluorescence spectrometer indicated that GO accelerated the renewal of bound EPS (B-EPS). Fourier Transform infrared spectroscopy (FTIR) results showed that GO affected the secondary structure of the protein in B-EPS, making B-EPS more hydrophobic and promoting microbial aggregation. B-EPS affected by GO can promote the electron transfer process of microorganisms. However, high concentration (>25 mg/L) of GO may inhibit denitrification by competing for electrons, which was not conducive to denitrification thermodynamically.

Microbial coupling mechanisms of nitrogen removal in constructed wetlands: A review.

Nitrogen removal through microorganisms is the most important pathway in constructed wetlands (CWs). In this review, we summarize the microbial coupling mechanisms of nitrogen removal, which are the common methods of nitrogen transformation. The electron pathways are shortened and consumption of oxygen and energy is reduced during the coupling of nitrogen transformation functional microorganisms. The highly efficient nitrogen removal mechanisms are cultivated from the design conditions in CWs, such as intermittent aeration and tidal flow. The coupling of microorganisms and substrates enhances nitrogen removal mainly by supplying electrons, and plants affect nitrogen transformation functional microorganisms by the release of oxygen and exudates from root systems as well as providing carriers for microbial attachment. In addition, inorganic elements such as Fe, S and H act as electron donors to drive the autotrophic denitrification process in CWs.

A high precision, fast response, and low power consumption in situ optical fiber chemical pCO2 sensor.

A sensor system suitable for monitoring changes in partial pressure of carbon dioxide (pCO(2)) in surface seawater or in the atmosphere has been developed. Surface seawater samples are pumped into a PVC tube enclosing an inner Teflon AF tube, which served as a long pathlength gas-permeable liquid-core waveguide for spectrophotometry. The Teflon cell contains a pH-sensitive indicator-buffer solution consisting of bromothymol blue (BTB) and sodium carbonate. Carbon dioxide in the sample diffuses into the indicator-buffer solution to reach equilibrium, resulting in pH changes, which are detected by changes in the absorbance of BTB at wavelengths of 620 and 434 nm. The pCO(2) in the sample is then derived from the pH change. The sensor has a response time of 2 min at the 95% equilibrium value and a measurement precision of 0.26-0.37% in the range 200-800 microatm pCO(2). This chemical sensor takes advantage of a combination of long pathlength, multiple wavelength detection, indicator solution renewal, and in situ automatic control technology, and has the feature of low power consumption (the average being approximately 4 W with a peak of approximately 8 W).