Rice consumption is one of the primary sources of arsenic (As) exposure as the grains contain relatively higher concentration of inorganic As. Abundant studies on the ability of iron (Fe) plaque in hampering As uptake by plants has been reported earlier. However, little is known about its role in the mitigation of As mediated oxidative damage in rice plants. The present study highlights the effect of As and Fe co-supplementation on growth response, oxidative stress, Fe uptake related enzymes and nutrient status in rice varieties. Eight different Indica rice varieties were screened and finally four varieties (Varsha, Jaya, PB-1 and IR-64) were selected for detailed investigations. Improved germination and chlorophyll/protein levels during As+Fe co-exposure indicate healthier plants than As(III) treated ones. Interestingly Fe was found act both as an antagonist and also as a synergist of As treatments. It acted by reducing As translocation and improving the nutritional levels and enhancing the oxidative stress. Fe uptake related enzymes (nitrite reductase and ferric chelate reductase) and phytosiderophores analysis revealed that Fe supplementation can reduce its deficiency in rice plants. Morpho-biochemical, oxidative stress and nutrient analysis symbolizes higher tolerance of PB-1 towards As, while Varsha being most sensitive, efficiently combated the As(III) stress in the presence of Fe.

The performance, nitrogen removal rate, microbial enzymatic activity and extracellular polymeric substances (EPS) of activated sludge were assessed under nickel (Ni(II)) stress. The organic matter and NH₄⁺-N removal efficiencies were stable at less than 10 mg/L Ni(II) and subsequently decreased with the increment of Ni(II) concentration from 10 to 30 mg/L. The specific oxygen uptake rate and dehydrogenase activity kept stable at less than 5 mg/L Ni(II) and then declined at 5–30 mg/L Ni(II). Both specific ammonia-oxidizing rate (SAOR) and specific nitrite-oxidizing rate (SNOR) decreased with the increment of Ni(II) concentration. The changing trends of ammonia monooxygenase and nitrite oxidoreductase activities were matched those of SAOR and SNOR, respectively. The nitrite-reducing rate and nitrate-reducing rate illustrated a similar variation tendency to the nitrite reductase activity and nitrate reductase activity, respectively. Ni(II) impacted on the production, chemical composition and functional group of EPS. The relation between the sludge volume index and the EPS production exhibited a better linear function with a negative slope, demonstrating that Ni(II) improved the sludge settleability despite of the increase of EPS production.

Benzylalkyldimethylethyl ammonium compounds are pervasive in natural environments and toxic at high concentrations. The changes in functional genes and microbial diversity in eutrophic lake samples exposed to benzyldimethyldodecyl ammonium chloride (BAC) were assessed. BAC exerted negative effects on bacteria abundance, particularly at concentrations of 100 μg L−1 and higher. A significant increase in the number of the quaternary ammonium compound-resistant gene qacA/B was recorded within the 10 μg L−1 treatment after the first day of exposure. Not all antibiotic resistance genes increased in abundance as the concentrations of BAC increased; rather, gene abundances were dependent on the gene type, concentrations of BAC, and contact time. The nitrogen fixation-related gene nifH and ammonia monooxygenase gene amoA were inhibited by high concentrations of BAC after the first day, whereas an increase of the nitrite reductase gene nirK was stimulated by exposure. Microbial communities within higher treatment levels (1000 and 10 000 μg L−1) exhibited significantly different community composition compared to other treatment levels and the control. Selective enrichment of Rheinheimera, Pseudomonas, and Vogesella were found in the higher treatment levels, suggesting that these bacteria have some resistance or degradation capacity to BAC. Genes related with RNA processing and modification, transcription, lipid transport and metabolism, amino acid transport and metabolism, and cell motility of microbial community function were involved in the process exposed to the BAC stress.

This study investigated the effects of arsenic on the in vitro activities of the enzymes (nitrate reductase and nitrite reductase) involved in nitrate metabolism in the roots, rhizomes, and fronds of four-month old Pteris vittata (arsenic - hyperaccumulator) and Pteris ensiformis (non-arsenic--hyperaccumulator) plants. The arsenic treatments (0, 150, and 300 μM as sodium arsenate) in hydroponics had adverse effects on the root and frond dry weights, and this effect was more evident in P. ensiformis than in P. vittata. Nitrate reductase and nitrite reductase activities of arsenate-treated plants were reduced more in P. ensiformis than in P. vittata. This effect was accompanied by similar decreases in tissue NO₃⁻ concentrations. Therefore, this decrease is interpreted as being indirect, i.e., the consequence of the reduced NO₃⁻ uptake and translocation in the plants. The study shows the difference in the tolerance level of the two Pteris species with varying sensitivity to arsenic. Arsenic reduced the activity of nitrate and nitrite reductase more in Pteris ensiformis than Pteris vittata.

Soil antibiotic resistome and the nitrogen cycle are affected by florfenicol addition to manured soils but their interactions have not been fully described. In the present study, antibiotic resistance genes (ARGs) and nitrogen cycle genes possessed by soil bacteria were characterized using real-time fluorescence quantification PCR (qPCR) and metagenomic sequencing in a short-term (30 d) soil model experiment. Florfenicol significantly changed in the abundance of genes conferring resistance to aminoglycosides, β-lactams, tetracyclines and macrolides. And the abundance of Sphingomonadaceae, the protein metabolic and nitrogen metabolic functions, as well as NO reductase, nitrate reductase, nitrite reductase and N₂O reductase can also be affected by florfenicol. In this way, ARG types of genes conferring resistance to aminoglycosides, β-lactamases, tetracyclines, colistin, fosfomycin, phenicols and trimethoprim were closely associated with multiple nitrogen cycle genes. Actinobacteria, Chlorobi, Firmicutes, Gemmatimonadetes, Nitrospirae, Proteobacteria and Verrucomicrobia played an important role in spreading of ARGs. Moreover, soil physicochemical properties were important factors affecting the distribution of soil flora. This study provides a theoretical basis for further exploration of the transmission regularity and interference mechanism of ARGs in soil bacteria responsible for nitrogen cycle.

The impact of antibiotics on denitrification has attracted widespread attention recently. Norfloxacin, as a representative of fluoroquinolone antibiotics, is extensively detected in groundwater. However, whether the release of norfloxacin into the groundwater poses potential risks to denitrification remains unclear. In this study, effect of norfloxacin on denitrification was investigated. The results showed that increasing norfloxacin from 0 to 100 μg/L decreased nitrate removal rate from 0.68 to 0.44 mg/L/h, but enhanced N₂O emission by 177 folds. Additionally, 100 μg/L of norfloxacin decreased nitrite accumulation by 50.6%. Corresponding inhibition of norfloxacin on bacterial growth, carbon source utilization, electron transport system activity and genes expression was revealed. Furthermore, denitrifying enzyme dynamic monitoring results showed that norfloxacin inhibited nitrate reductase activity, and enhanced nitrite reductase activity to some extent in denitrification process, which was consistent with the variations of nitrate and nitrite. Meanwhile, sensitivity analysis demonstrated that nitrate reductase was more easily affected by norfloxacin than nitrite reductase. Overall, this study suggests that multiple regulation of denitrifying enzyme activity contributes to evaluating the comprehensive effects of antibiotics on groundwater denitrification.

Rhizospheric microorganisms such as denitrifying bacteria are able to affect ‘rhizobioaugmention’ in aquatic plants and can help boost wastewater purification by benefiting plant growth, but little is known about their effects on the production of plant root exudates, and how such exudates may affect microorganismal nitrogen removal. Here, we assess the effects of the rhizospheric Pseudomonas inoculant strain RWX31 on the root exudate profile of the duckweed Spirodela polyrrhiza, using gas chromatography/mass spectrometry. Compared to untreated plants, inoculation with RWX31 specifically induced the exudation of two sterols, stigmasterol and β-sitosterol. An authentic standard assay revealed that stigmasterol significantly promoted nitrogen removal and biofilm formation by the denitrifying bacterial strain RWX31, whereas β-sitosterol had no effect. Assays for denitrifying enzyme activity were conducted to show that stigmasterol stimulated nitrogen removal by targeting nitrite reductase in bacteria. Enhanced N removal from water by stigmasterol, and a synergistic stimulatory effect with RWX31, was observed in open duckweed cultivation systems. We suggest that this is linked to a modulation of community composition of nirS- and nirK-type denitrifying bacteria in the rhizosphere, with a higher abundance of Bosea, Rhizobium, and Brucella, and a lower abundance of Rubrivivax. Our findings provide important new insights into the interaction of duckweed with the rhizospheric bacterial strain RWX31 and their involvement in the aquatic N cycle and offer a new path toward more effective bio-formulations for the purification of N-polluted waters.

Biochar may variably impact nitrogen (N) transformation and N-cycle-related microbial activities. Yet the mechanism of biochar amendment on nitrous oxide (N₂O) emissions from agricultural ecosystems remains unclear. Based on a 6-year long-term biochar amendment experiment, we applied a dual isotope (¹⁵N–¹⁸O) labeling technique with tracing transcriptional genes to differentiate the contribution of nitrifier nitrification (NN), nitrifier denitrification (ND), nitrification-coupled denitrification (NCD) and heterotrophic denitrification (HD) pathway to N₂O production. Then the field experiment provided quantitative data on dynamic N₂O emissions, soil mineral N and key functional marker gene abundances during the wheat growing season. By using ¹⁵N–¹⁸O isotope, biochar decreased N₂O emission derived from ND (by 45–94%), HD (by 35–46%) and NCD (by 30–64%) compared to the values under N application. Biochar increased the relative contribution of NN to total N₂O production as evidenced by the increase in ammonia-oxidizing bacteria, but did not influence the cumulative NN-derived N₂O. The field experiment found that the majority of the N₂O emissions peaked following fertilization, in parallel with soil NH₄⁺ and nitrite dynamics. Soil N₂O emissions during the wheat growing stage were effectively decreased (by 38–48%) by biochar amendment. Based on the correlation analyses and random forest analysis in both microcosm and field experiments, the decrease in nitrite concentration (by 62–65%) and increase in N₂O consumption were mainly responsible for net N₂O mitigation, as evidenced by the decrease in the ratios of nitrite reductase genes/transcripts (nirS, nirK and fungal nirK) and N₂O reductase gene/transcripts (nosZI and nosZII). Based on the extrapolation from microcosm to field, biochar significantly mitigated N₂O emissions by weakening the ND processes, since NCD and HD contributed little during the N₂O emission “peaks” following urea fertilization. Therefore, emphasis should be put on the ND process and nitrite accumulation during N₂O emission peaks and extrapolated to all agroecosystems.

The stability of community functioning in anaerobic ammonia oxidation (anammox) sludge adaptation to various salinity changes are concerned but not fully explored. In this study, two anammox reactors were designed in response to different salt levels and salt-adding methods. The reactor PI, run with small stepwise salt increments (0.5%–1.0%), removed >90% of nitrite and ammonium in the influent over the range of 0%–4% salt. By contrast, the reactor SI, run with a sharp salt increment (>2.5%), exhibited a reduced performance (by up to 44%) over the same salt range with a new steady state. The observed resilience times after salt perturbations indicated that the PI reactor recovered substantially and rapidly at all imposed salt levels. Principal coordinates analysis of 16S rRNA gene amplicon sequences revealed that bacterial community structures of the anammox sludge altered conspicuously in response to the salinity changes. However, quantitative PCR analysis showed that the shift in copy number of studied nitrogen-converting genes encoding hydrazine synthase (hzsA), bacterial and archaeal ammonia monooxygenases (amoA), nitrite oxidoreductase (nxrB), nitrite reductase (nirK), and nitrous oxide reductase (nosZ) was not significant (p > 0.05) in anammox sludge across the salt levels of 0.5%–4%, which suggests the stability of microbial community functioning in the osmoadaptation processes. The freshwater anammox Ca. Kuenenia showed high osmoadaptation by potentially adopting both high-salt-in and low-salt-in strategies to dominate in both reactors. The quantitative transcript analysis showed that the active anammox bacteria represented by hzsA transcripts in the SI reactor were approximately two orders of magnitude lower than those in the PI reactor during the long-term exposure to 4% salinity, manifesting the influence by the salt-increasing methods. These results provided new insight into osmo-adaptation of the anammox microbiome and will be useful for managing salinity effects on nitrogen removal processes.

In the present study, we investigated the influence of phenanthrene (PHE), a three-ring polycyclic aromatic hydrocarbon (PAH) compound, on nitrate (NO3−) reduction processes in mangrove sediments using microcosms. After 10days, nitrate/nitrite reductase activity and abundance of narG and nirS significantly decreased in the bulk sediment at both 10/50mgPHEkg−1 contamination groups. In the rhizosphere, abundance of narG, nirS and nirK markedly declined at PHE treated sediments, while the drop in reductase activity at 10mgkg−1 PHE treatment was insignificant. After 50days, apart from 10mgPhekg−1 treated bulk sediment, abundance of denitrifiers and reductase activity in all PHE spiked sediment samples significantly dropped. Therefore, the influence of PAHs on NO3− reduction capability in mangrove sediments is dependent on spiked concentration, temporal scale of exposure and interaction with roots. Generally, PAHs play an inhibitor role, slowing NO3− turnover rates, which warrant attention from coastal managers.