Are nitrogen and carbon cycle processes impacted by common stream antibiotics? A comparative assessment of single vs. mixture exposures.

A variety of antibiotics are ubiquitous in all freshwater ecosystems that receive wastewater. A wide variety of antibiotics have been developed to kill problematic bacteria and fungi through targeted application, and their use has contributed significantly to public health and livestock management. Unfortunately, a substantial fraction of the antibiotics applied to humans, pets and livestock end up in wastewater, and ultimately many of these chemicals enter freshwater ecosystems. The effect of adding chemicals that are intentionally designed to kill microbes, on freshwater microbial communities remains poorly understood. There are reasons to be concerned, as microbes play an essential role in nutrient uptake, carbon fixation and denitrification in freshwater ecosystems. Chemicals that reduce or alter freshwater microbial communities might reduce their capacity to degrade the excess nutrients and organic matter that characterize wastewater. We performed a laboratory experiment in which we exposed microbial community from unexposed stream sediments to three commonly detected antibiotics found in urban wastewater and urban streams (sulfamethoxazole, danofloxacin, and erythromycin). We assessed how the form and concentration of inorganic nitrogen, microbial carbon, and nitrogen cycling processes changed in response to environmentally relevant doses (10 µg/L) of each of these antibiotics individually and in combination. We expected to find that all antibiotics suppressed rates of microbial mineralization and nitrogen transformations and we anticipated that this suppression of microbial activity would be greatest in the combined treatment. Contrary to our expectations we measured few significant changes in microbially mediated functions in response to our experimental antibiotic dosing. We found no difference in functional gene abundance of key nitrogen cycling genes nosZ, mcrA, nirK, and amoA genes, and we measured no treatment effects on NO3- uptake or N2O, N2, CH4, CO2 production over the course of our seven-day experiment. In the mixture treatment, we measured significant increases in NH4+ concentrations over the first 24 hours of the experiment, which were indistinguishable from controls within six hours. Our results suggest remarkable community resistance to pressure antibiotic exposure poses on naïve stream sediments.

Stability and Microbial Toxicity of Silver Nanoparticles under Denitrifying Conditions.

While the antibacterial effect of silver nanoparticles (AgNPs) on environmentally beneficial microbes has drawn considerable attention, the stability and microbial toxicity of AgNPs in a system where nitrate reduction is the dominant terminal electron-accepting process remain understudied. Here, we explore the impact of citrate-coated AgNPs (cit-AgNPs) on the growth and metabolism of two metal-sensitive and one nonsensitive bacterial strains under denitrifying conditions. Dose-response analysis revealed that in contrast to the bacteriostatic effect exhibited at 1 ppm, 5 ppm cit-AgNPs were bactericidal to the metal-sensitive strains. It was observed that the growth of the cells initiated Ag(I) formation, and the supplement of chloride (2.7 mM) to the cultures substantially mitigated the bactericidal capacity of cit-AgNPs, indicating that AgNP dissolution to ionic Ag(I) played a key role in AgNP toxicity. Abiotic experiments confirmed that nitrite, not nitrate, had the capacity to oxidize cit-AgNPs. Transcriptomic analysis revealed that (i) the gene encoding for membrane stress was upregulated proportionally to cit-AgNP concentrations; (ii) cit-AgNPs and Ag(I) at higher levels upregulated genes involved in oxidative stress and iron-sulfur clusters, whereas expressions of the genes responsible for electron transport, ATP synthesis, and denitrification were substantially repressed; (iii) the addition of chloride significantly altered the level of transcriptional profiles of all of the genes. These results not only provide evidence of abiotic AgNP oxidation by metabolic intermediate nitrogen species but also suggest that AgNPs and Ag(I) may induce differential toxicity modes to prokaryotes. Our findings reinforce the importance of evaluating the potential ecological toxicity and risks associated with the transformation of nanomaterials.

Response characteristics of nirS-type denitrifier Paracoccus denitrificans under florfenicol stress.

Florfenicol (FF) is widely used in aquaculture and can interfere with denitrification when released into natural ecosystems. The aim of this study was to analyze the response characteristics of nirS-type denitrifier Paracoccus denitrificans under FF stress and further mine antibiotic-responsive factors in aquatic environment. Phenotypic analysis revealed that FF delayed the nitrate removal with a maximum inhibition value of 82.4% at exponential growth phase, leading to nitrite accumulation reached to 21.9-fold and biofilm biomass decreased by ~38.6%, which were due to the lower bacterial population count (P < 0.01). RNA-seq transcriptome analyses indicated that FF treatment decreased the expression of nirS, norB, nosD and nosZ genes that encoded enzymes required for NO2- to N2 conversion from 1.02- to 2.21-fold (P < 0.001). Furthermore, gene associated with the flagellar system FlgL was also down-regulated by 1.03-fold (P < 0.001). Moreover, 10 confirmed sRNAs were significantly induced, which regulated a wide range of metabolic pathways and protein expression. Interestingly, different bacteria contained the same sRNAs means that sRNAs can spread between them. Overall, this study suggests that the denitrification of nirS-type denitrifiers can be hampered widely by FF and the key sRNAs have great potential to be antibiotic-responsive factors.

Effects of sulfamethoxazole on nitrogen removal and molecular ecological network in integrated vertical-flow constructed wetland.

Response of nitrogen removal efficiency and microbial interactions to organic pollution has been a major issue in wastewater treatment system. However, the nitrogen removal efficiency and interactions among microbial community under antibiotics press is still unclear. Thus, the effect of sulfamethoxazole (SMX) on nitrogen removal and microbial responses of IVCWs was investigated through recorded the nitrogen removal efficiency before and after adding SMX and random matrix theory (RMT)-based network analysis. Results showed that better NH4+-N removal (>90%) after a long period of operation were achieved in IVCWs, but NO3--N was accumulated. However, nitrate removal rates were significantly increased after long-term exposure (60 d) to 100 µgL-1 SMX (from 27.35% to 35.57%) with relatively high SMX removal (53.50%). Surprisingly, the ammonia nitrogen removal rate (90.07-92.70%) were not significantly affected by SMX in IVCWs. Moreover, the bacterial richness was decreased and the bacterial community structures were altered by the presence of SMX, especially those of nitrogen-transforming microorganisms. Molecular ecological network analysis suggested that SMX had positive influences on denitrifying bacteria interactions but reduced the network complexity and microbial interactions on whole molecular network, and among-module connections were weakened obviously at SMX.

Effects of copper and florfenicol on nirS- and nirK-type denitrifier communities and related antibiotic resistance in vegetable soils.

Denitrification play an important role in nitrogen cycle and is affected by veterinary drugs entering agricultural soils. In the present study, the effects of copper and florfenicol on denitrification, related antibiotic resistance and environmental variables were characterized using real-time quantitative PCR (qPCR) and amplicon sequencing in a short-term (30 d) soil model experiment. Drug additions significantly decreased the nirS gene abundance (P < 0.05) but maximized the abundance of gene nirK in soil containing florfenicol and moderate copper levels (150 mg kg-1). Surprisingly, copper additions decreased the fexB gene abundance, however, the abundance of gene pcoD significantly increased in soils containing florfenicol, moderate copper levels (150 mg kg-1), and florfenicol and low copper levels (30 mg kg-1), respectively (P < 0.05). Overall, the nirK-type community composition was more complex than that of nirS-type but Proteobacteria predominated (> 90%) in both communities. Correlation analysis indicated that the gene abundance of fexB was highly correlated with NH4+-N (P < 0.05) and NO3--N (P < -0.01), and floR gene abundance was positively correlated with nirK (P < 0.01). Besides, the abundance of nirS-type genera Bradyrhizobium and Pseudomonas were obviously related to total organic matter (TOM), total nitrogen (TN) or total phosphorus (TP) (P < 0.05), while the abundance of nirK-type Rhizobium, Sphingomonas and Bosea showed a significantly correlated with TOM, TN or copper contents (P < 0.05). Taken together, copper and florfenicol contamination increased the possibility of durg resistance genes spread in agricultural soils through nitrogen transformation.

Antibiotic-induced role interchange between rare and predominant bacteria retained the function of a bacterial community for denitrifying quinoline degradation.

AIM: Quinoline is a recalcitrant pollutant in coking wastewater which has been broadly investigated with many isolates possessing aerobic quinoline-degrading ability. However, studies on anaerobic degradation and the corresponding bacteria are very scarce. This study attempted to investigate the role of diverse functional members and the redundancy of quinoline degradation in a lab-scale quinoline denitrifying bioreactor. METHODS AND RESULTS: Antibiotics were added to the batch culture under denitrifying conditions to disturb the microbial community of the quinoline-degrading bioreactor. According to the results, the nitrate removal rate remained stable, and the quinoline removal rate increased by 9·7% after treatment with streptomycin. However, PCoA analysis of 16S rRNA gene sequencing data of these samples indicated a significant shift in microbial community structures. Specifically, 12 operational taxonomic units (OTUs), including OTU1 (Pseudomonas) and OTU2 (Achromobacter), were significantly enriched. OTU1 replaced OTU8 (Thauera) as the most predominant denitrifying quinoline-degrading member. However, OTU8 and other predominant OTUs (Comamonas and Pseudoxanthomonas), which were hypothesized to contribute essentially to quinoline degradation in the origin bioreactor, became almost undetectable. CONCLUSION: Functional redundancy due to high biological diversity allowed the role reversal of predominant quinoline-degrading bacteria and other rare bacteria when disturbed by antibiotic stress. Although the abundance of OTU1 was much lower initially, it replaced the essential role of the predominant member OTU8 in the bioreactor community for quinoline degradation once the environmental condition changed. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicated that the high biological diversity in a wastewater treatment bacterial community is crucial for maintaining the degrading function of organic pollutants, especially in a changing environment due to external disturbance or stress.

New application of rutin: Repair the toxicity of emerging perfluoroalkyl substance to Pseudomonas stutzeri.

Per- and polyfluoroalkyl substances (PFASs) are toxic to microorganisms, thereby affecting microbial communities in sludge and soil, but how to repair the toxicity of microorganisms remains unclear. In this study, rutin, an antioxidant, was added into a culture medium with an aerobic denitrification bacteria, Pseudomonas stutzeri, under the exposure of sodium perfluorononyloxy-benzenesulfonate (OBS) to evaluate the repair mechanisms of rutin to the toxicity of OBS to the bacteria. The results showed that rutin could repair the damage of OBS to cell structures, and reduce the death rates of the bacteria under OBS exposure. The dosage of rutin reduced the effect on the inhibition of denitrification ability of P. stutzeri under OBS exposure. Compared with the bacteria exposed to single OBS, the dosage of rutin resulted in that the death rates recovered from 96.2% to 66.4%, the growth inhibition rate decreased from 46.5% to 15.8%, the total nitrogen removal rate recovered from 66.9% to 100%, and the NO2- content recovered from 34.5 mg/L to 0.22 mg/L. The expressions of key denitrification genes (napA, nirS, norB, nosZ) were recovered after adding rutin under OBS exposure. Rutin changed the positive rate of reactive oxygen species, the relative superoxide dismutase and catalase activities in the bacteria which exposed to OBS. The mechanism by which rutin repaired the toxicity of OBS to P. stutzeri related to inhibiting the activities of antioxidant and denitrification enzymes rather than affecting the expressions of genes involved in these enzymes. This study sheds light on the repair method of micro-organics and reveals the repair mechanisms under PFASs exposure.

Microplastics affect sedimentary microbial communities and nitrogen cycling.

Microplastics are ubiquitous in estuarine, coastal, and deep sea sediments. The impacts of microplastics on sedimentary microbial ecosystems and biogeochemical carbon and nitrogen cycles, however, have not been well reported. To evaluate if microplastics influence the composition and function of sedimentary microbial communities, we conducted a microcosm experiment using salt marsh sediment amended with polyethylene (PE), polyvinyl chloride (PVC), polyurethane foam (PUF) or polylactic acid (PLA) microplastics. We report that the presence of microplastics alters sediment microbial community composition and nitrogen cycling processes. Compared to control sediments without microplastic, PUF- and PLA-amended sediments promote nitrification and denitrification, while PVC amendment inhibits both processes. These results indicate that nitrogen cycling processes in sediments can be significantly affected by different microplastics, which may serve as organic carbon substrates for microbial communities. Considering this evidence and increasing microplastic pollution, the impact of plastics on global ecosystems and biogeochemical cycling merits critical investigation.

Influences of organic nitrogen on the removal of inorganic nitrogen from complicated marine aquaculture water by Marichromatium gracile YL28.

The sediment-water interface is not only an important location for substrate conversion in a mariculture system, but also a major source of eutrophication. This study aimed to clarify the characteristics of inorganic nitrogen (ammonia, nitrite and nitrate) removal by Marichromatium gracile YL28 in the presence of both organic nitrogen and inorganic nitrogen. The results showed that, in the presence of peptone or urea, seaweed oligosaccharides (SOS) effectively enhanced the ammonia removal capacity of YL28 (6.42 mmol/L) and decreased the residual rate by 54.04% or 8.17%, respectively. With increasing peptone or urea concentrations, the removal of both ammonia and nitrate was gradually inhibited, and the residual rates of ammonia and nitrate reached 22.56-34.36% and 12.03-15.64% in the peptone system and 20.65-24.03% and 12.20-13.21% in the urea system, respectively. However, in the control group the residual rates of ammonia and nitrate reached 11.97% and 5.12%, respectively. In addition, the concentrations of peptone and urea did not affect nitrite removal, and YL28 displayed better cell growth and nitrogen removal activity in the presence of bait and SOS. Overall, the ability of YL28 to remove inorganic nitrogen was enhanced in the presence of organic nitrogen.

Transcriptional and metabolic response against hydroxyethane-(1,1-bisphosphonic acid) on bacterial denitrification by a halophilic Pannonibacter sp. strain DN.

Biological denitrification is an environmentally sound pathway for the elimination of nitrogen pollution in wastewater treatment. Extreme environmental conditions, such as the co-existence of toxic organic pollutants, can affect biological denitrification. However, the potential underlying mechanism remains largely unexplored. Herein, the effect of a model pollutant, hydroxyethane-(1,1-bisphosphonic acid) (HEDP), a widely applied and consumed bisphosphonate, on microbial denitrification was investigated by exploring the metabolic and transcriptional responses of an isolated denitrifier, Pannonibacter sp. strain DN. Results showed that nitrate removal efficiency decreased from 85% to 50% with an increase in HEDP concentration from 0 to 3.5 mM, leading to nitrite accumulation of 204 mg L-1 in 3.5 mM HEDP. This result was due to the lower bacterial population count and reduction in the live cell percentage. Further investigation revealed that HEDP caused a decrease in membrane potential from 0.080 ±â€¯0.005 to 0.020 ±â€¯0.002 with the increase in HEDP from 0 to 3.5 mM. This hindered electron transfer, which is required for nitrate transformation into nitrogen gas. Moreover, transcriptional profiling indicated that HEDP enhanced the genes involved in ROS (O2-) scavenging, thus protecting cells against oxidative stress damage. However, the suppression of genes responsible for the production of NADH/FADH2 in tricarboxylic acid cycle (TCA), NADH catalyzation (NADH dehydrogenase) in (electron transport chain) ETC system and denitrifying genes, especially nor and nir, in response to 2.5 mM HEDP were identified as the key factor inhibiting transfer of electron from TCA cycle to denitrifying enzymes through ETC system.

Effects of silver nanoparticles on coupled nitrification-denitrification in suspended sediments.

The effects of varying concentrations of Ag NPs on coupled nitrification and denitrification (CND) in two suspended sediments (SPSs) sizes were investigated using isotopic tracer method. In general, 0.5 and 5 mg/L Ag NPs had less effect on CND, while 2 and 10 mg/L Ag NPs exhibited the improvement and inhibition effect, respectively. The CND improvement by 2 mg/L NPs was mainly due to the enhanced nitrifying and denitrifying enzyme activity. However, 10 mg/L Ag NPs inhibited NH4+ oxidation by directly reducing the AMO activity and AOB abundance. The inhibition on NAR and NIR activity and their encoding narG and nirK gene abundance further inhibited NO3- and NO2- reduction, leading to a dramatic decrease in the 15N-N2 production. The above inhibition effects were attributed to the nano-effects of Ag NPs, which led to the excessive ROS amount and the decreased T-AOC level in microbial systems. But the connection between nitrification and denitrification was not broken after Ag NPs exposure. Moreover, the results indicated that N-cycling in clay and silt-type SPS systems could be more sensitive than sand-type SPS systems to NP exposure. The findings provide a basis for evaluating the environmental risks of Ag NPs in water-sediment systems.

Copper oxide nanoparticles inhibited denitrifying enzymes and electron transport system activities to influence soil denitrification and N2O emission.

Nanopesticides are widely applied in modern agricultural systems to replace traditional pesticides, which inevitably leads to their accumulation in soils. Nanopesticides based on copper oxide nanoparticles (CuO NPs) may affect the soil nitrogen cycle, such as the denitrification process; however, the mechanism remains unclear. Here, acute exposure experiments for 60 h were conducted to explore the effects of CuO NPs (10, 100, 500 mg kg-1) on denitrification. In this study, Cu speciation, activities of denitrifying enzymes, electron transport system activity (ETSA), expression of denitrifying functional genes, composition of bacterial communities and reactive oxygen species (ROS) were determined. In all treatments, Cu ions was the dominant form and responsible for the toxicity of CuO NPs. The results indicated that CuO NPs treatments at 500 mg kg-1 remarkably inhibited denitrification, led to an 11-fold increase in NO3- accumulation and N2O emission rates decrease by 10.2-24.1%. In the denitrification process, the activities of nitrate reductase and nitric oxide reductase reduced by 21.1-42.1% and 10.3-16.3%, respectively, which may be a reason for the negative effect of CuO NPs. In addition, ETSA was significantly inhibited with CuO NPs applications, which reflects the ability of denitrification to accept electrons. Denitrifying functional genes and bacterial communities composition were changed, thus further influencing the denitrification process. ROS analysis showed that there were no significant differences among NPs treatments. This research improves the understanding of CuO NPs impact on soil denitrification. Further attention should be paid to the nitrogen transformation in agricultural soils in the presence of nanopesticides.

Effects of carbon nanotube on denitrification performance of Alcaligenes sp. TB: Promotion of electron generation, transportation and consumption.

Multi-walled carbon nanotubes (MWCNTs) promote biodegradation in water treatment, but the effect of MWCNT on denitrification under aerobic conditions is still unclear. This investigation focused on the denitrification performance of MWCNT and its toxic effects on Alcaligenes sp. TB which showed that 30 mg/L MWCNTs increased NO3- removal efficiency from 84% to 100% and decreased the NO2-and N2O accumulation rates by 36% and 17.5%, respectively. Nitrite reductase and nitrous oxide reductase activities were further increased by 19.5% and 7.5%, respectively. The mechanism demonstrated that electron generation (NADH yield) and electron transportation system activity increased by 14.5% and 104%, respectively. Cell membrane analysis found that MWCNT caused an increase in polyunsaturated fatty acids, which had positive effects on electron transportation and membrane fluidity at a low concentration of 96 mg/kg but caused membrane lipid peroxidation and impaired membrane integrity at a high concentration of 115 mg/L. These findings confirmed that MWCNT affects the activity of Alcaligenes sp. TB and consequently enhances denitrification performance.

Influence of chlorothalonil and carbendazim fungicides on the transformation processes of urea nitrogen and related microbial populations in soil.

To improve crop yielding, a large amount of fungicides is continuously applied during the agricultural management, while the effects of fungicides residues on microbial processing of N in soil need further study. In the present study, two broad spectrum fungicides, chlorothalonil and carbendazim, were applied at the rates of 5, 10, and 50 mg of active ingredient (A.I.) per kg of dry soil combined with urea with 200 mg of N per kg of dry soil under laboratory conditions. The results showed that chlorothalonil obviously retarded the hydrolysis of urea, whereas carbendazim accelerated it in 4 days after the treatments (P < 0.05). Chlorothalonil reduced denitrification, nitrification, and N2O production (P < 0.05), but not for carbendazim. Further analysis on N-associated microbial communities showed chlorothalonil reduced nitrosomonas populations at the rates of 10 and 50 mg of A.I. per kg and autotrophic nitrifying bacterial populations at three application rates (P < 0.05), but Carbendazim decreased nitrosomonas populations only at the rate of 50 mg of A.I. per kg and also autotrophic nitrifying bacterial populations at three rates and heterotrophic nitrifying bacterial populations at the rates of 10 and 50 mg of A.I. per kg. The reasons for this difference were ascribed to arrest urea hydrolysis and impediment of denitrification and nitrification processes by chlorothalonil. In conclusion, to improve crop yielding, chlorothalonil might be more beneficial to conserve soil N by improving soil N fertility, compared with carbendazim.

Tetracycline-induced effects on the nitrogen transformations in sediments: Roles of adsorption behavior and bacterial activity.

Nitrification and denitrification are the most important nitrogen transformation processes in the environment. Recently, due to widespread use, antibiotics have been reported to lead to environmental risks. Tetracycline (TC) is one of the most extensively used antibiotics in many areas. However, its reported effects on nitrogen transformations were conflicting in previous studies. In this study, the effects of TC on nitrogen transformations in sediment were investigated by analyzing TC transport and bacterial activity. It was found that the adsorption of TC onto the sediment was favorable and spontaneous, with adsorption capacity 54.3 mg/kg. The adsorption kinetics of TC onto the sediment and the isotherm fitted the Elvoich and Freundlich models, respectively, indicating that the adsorption was a chemisorption process, including electrostatic interactions and chemical bonding between TC and the sediment. TC showed no effect on nitrification in the sediment, but significantly inhibited the reduction of nitrate and nitrite during denitrification, consistent with observations made for the model denitrifier Paracoccus denitrificans under TC stress. Mechanistic study indicated that TC at 130 µg/g-cell inhibited 50.7% of P. denitrificans growth and 61.6% of cell viability. Meanwhile, the catalytic activities of the key denitrifying enzymes, nitrate reductase (NAR) and nitrite reductase (NIR), decreased to 29.1% and 68.0% of the control levels when the TC concentration was 130 µg/g-cell, suggesting that NAR was more sensitive to the TC than NIR, which contributed to a delay in nitrite accumulation.

Analysis for microbial denitrification and antibiotic resistance during anaerobic digestion of cattle manure containing antibiotic.

This study investigated the effects of tylosin (0, 10, and 100 mg/kg dry weight) on the denitrification genes and microbial community during the anaerobic digestion of cattle manure. N2 emissions were reduced and N2O emissions were increased by 10 mg/kg tylosin. Adding 100 mg/kg tylosin increased the emission of both N2O and N2. The different responses of denitrifying bacteria and genes to tylosin may have been due to the presence of antibiotic resistance genes (ARGs). Network analysis indicated that denitrification genes and ARGs had the same potential host bacteria. intI1 was more important for the horizontal transfer of denitrification genes and ARGs during anaerobic digestion than intI2. The anaerobic digestion of manure containing tylosin may increase nitrogen losses and the associated ecological risk.

Understanding the granulation of partial denitrification sludge for nitrite production.

Partial-denitrification (PD) has previously been demonstrated to be another pathway for nitrite production, which provides a cost-effective approach for nitrate (NO3--N) removal through combing with anammox. In this study, the formation of PD granules was firstly investigated in a sequencing batch reactor (SBR) with influent nitrate of 60 mg N/L. The granulation process was explored via the physicochemical and biological characterization. Sludge granulation initiated within the first 20 days with an average size of 93.7 µm in diameter, it experienced a developing, shaping and matured periods, with a maximum size of 709.3 µm obtained. High nitrite production of PD was always maintained during the granulation with a mean nitrate-to-nitrite transformation ratio (NTR) of 88.3%, and in-situ maximum NO3-N reduction rate of 84.9 mg N/h/g VSS was obtained. Mature PD granules hold an excellent settling property with 5-min sludge volume index (SVI5) of 32.0 mL/g MLSS obtained and smooth surface with large amounts of rod bacteria covered. CaCO3 precipitates formed in the PD process played a vital role in the initial granulation, acting as the nucleus for cell attachment. Extracellular polymeric substances (EPS), mainly the proteins (PN) content, was found to be of supreme importance in granules developing and maintaining its structural stability. Besides, the abundance of Flavobacterium and norank_p__Gracilibacteria were revealed to be in accordance with the change of granules size, seemed to contribute to sludge granulation. The developed granule-based PD integrated with anammox process provides an engineering-feasible and economic-favorable solution for industrial nitrate wastewater treatment.

Long-term impact of sulfate on an autotrophic nitrogen removal system integrated partial nitrification, anammox and endogenous denitrification (PAED).

A nitrogen removal system integrating partial nitrification, anaerobic ammonium oxidation (Anammox) and endogenous denitrification (PAED) was established in a sequencing batch reactor (SBR) for treating low nitrogen sewage (approximately 40 mg L-1 ammonia-nitrogen). The impact of sulfate on PAED sludge was investigated in five identical SBRs, fed with different levels of sulfate (0, 50, 100, 200 and 400 mg L-1). Ammonia oxidation was improved by the addition of Results showed that the sulfate addition in low concentration of sulfate (≤50 mg L-1), but was profoundly suppressed by higher levels of sulfate. Sulfate feeding enhanced both total nitrogen removal by Anammox and endogenous denitrification, with the abundance of Candidatus Kuenenia increasing to 4.39% in 400 mg L-1 sulfate from 0.83% in the control reactor, and Denitratisoma increasing to 6.35% from 2.77%. The results proved the feasibility of the PAED system in treating low nitrogen sewage with sulfate, which also enhanced the nitrogen-sulfate interaction.

Effect of plant-based carbon source supplements on denitrification of synthetic wastewater: focus on the microbiology.

The effects of plant-based carbon source addition on wastewater NO3--N removal and the involved microorganisms, especially denitrifying bacteria, were investigated. A synthetic wastewater (NO3--N, 15 mg/L) was treated through the batch experiment, which included three inoculation cycles (7 days/cycle), and was conducted at 25 °C. Four natural plant substrates, namely, rice straw (RS), wheat straw (WS), ryegrass (RG), and reed (RD), were used as carbon sources and supplemented at the rate of 1% (w/v). The results showed that both RS and WS performed well in promoting NO3--N removal (79.55-97.07%). While RG removed only 22.08% of NO3--N in the first cycle, the removal efficiency increased afterward (86.09-95.82%). Conversely, the NO3--N removal rate of RD decreased from 95.10 to 24.77% as a result of its low ability to supply carbon. With respect to the microorganisms, the RS treatment resulted in more bacteria and denitrifying genes such as narG, nirK, nirS, and norB than other treatments, while the highest number of nosZ gene copies was recorded in the WS treatment. Sequencing results revealed that Firmicutes (18.19-56.96%), Proteobacteria (38.82-74.80%), and Bacteroidetes (3.15-4.15%) were three dominant bacterial phyla for RS, WS, and RD treatments. Furthermore, the genera Enterobacter, Massilia, and Bacillus were the main denitrifying bacteria participating in the NO3--N removal. Furthermore, correlation analysis indicated that the denitrifying genus Sphingobacterium played an important role in enhancing nitrogen removal. This study suggested that RS is the superior plant-based carbon source for denitrifying bioreactors used in agricultural runoff treatment.

Does rice straw application reduce N2O emissions from surface flow constructed wetlands for swine wastewater treatment?

Rice straw was applied often as a carbon source to improve nitrogen removal; however, few studies have considered the effect of rice straw on nitrous oxide (N2O) emission during nitrogen removal in constructed wetlands (CWs). We constructed eighteen combined systems, consisting of rice straw ponds and surface flow CWs to investigate the effect of rice straw application on N2O emission in three strengths of swine wastewater treatments. The results showed rice straw (RS) treatment increased 131.5% of N2O emission factor from low strength CWs, but decreased 37.2-43.7% of N2O emission factors for medium and high strengths compared with no rice straw (NRS) treatment. The RS application led to an average 10.7% increase in the potential denitrification rate, and simultaneously enhanced gene abundances of the total bacteria (16S rRNA), ammonia-oxidising archaea, ammonia-oxidising bacteria, nitrate reductase, and N2O reductase (nosZ) for all strengths CWs. The multiple regression model revealed N2O emissions were strongly related to water temperature, nitrate, chemical oxygen demand, and denitrification genes. The proportion of nosZ gene abundance in 16S rRNA was higher in RS (0.7-1.3%) than NRS (0.4-0.9%) for medium and high strengths, while an opposite trend was observed for low strength. The discrepancy was responsible for increasing or decreasing N2O emission by RS application among different strengths. These findings indicated the effectiveness of RS application to control N2O emissions from the surface flow CWs was related to the pollution level of wastewater.