The agricultural sector uses fertilizers such as urea to add more nutrients to the soil needed for plant growth. Although it is cost-effective in crop production, indiscriminate use of nitrate-based fertilizer may result in behavioural, morphological, and physiological alterations on non-target organisms. This study determined the angiogenesis activity in the chorioallantoic membrane of urea-exposed duck embryos. It also investigated the weight, morphometries, and liver histopathology to gather more information on urea fertilizer's toxicity. It was observed that urea promoted angiogenesis in the CAM of duck embryos, especially at higher concentrations (P<0.05). Embryos treated with urea resulted in an alteration of the head-beak length (P<0.05). However, weight, crown-rump length, forelimb length, and hind limb length were not affected. The developing liver of urea-treated embryos showed distortion of the central vein shape and had larger sinusoidal spaces. The presence of Kupffer cells and lipid droplets were observed in the treated section. Congestion of blood cells, haemorrhage, and necrosis of hepatocytes were also observed in the tissue suggesting the extent of damage caused by the fertilizer. The findings of this study showed multiple developmental effects of urea on duck embryos. Further investigations are needed to shed more light on the toxicity of urea fertilizer on vertebrates.

Raw biochar with high pH possibly stimulated ammonia (NH₃) volatilization in the agricultural soil. We hypothesized that the modified biochar (MBC) with low pH can synchronically decrease the NH₃ and nitrous oxide (N₂O) losses. We performed a two-year experiment to clarify how citric acid MBC influence the NH₃ volatilization and N₂O emission as well as the underlying mechanisms. Two typical paddy soils, i.e., Hydragric Anthrosol and Haplic Acrisol, receiving equal urea N with 240 kg ha⁻¹ but varied rates of MBC with 0, 5, 10, and 20 t ha⁻¹ (named Urea, Urea + MBC5, Urea + MBC10, and Urea + MBC20, respectively) were studied. The results showed that MBC-amended treatments effectively mitigated the NH₃ volatilization from Hydragric Anthrosol and Haplic Acrisol by 29.6%–57.9% and 30.5%–62.4% in 2017, and by 16.5%–21.0% and 24.5%–35.0% in 2018, respectively, compared to Urea treatment. In addition, significantly lower N₂O emissions with averaged 38.3% and 43.1% in 2017, and 51.7% and 26.7% were recorded under Hydragric Anthrosol and Haplic Acrisol, respectively, following the MBC application (P < 0.05). Increased MBC addition performed higher efficacy on mitigating NH₃ volatilization, particularly in the first rice season, while this “dosage effect” was not found for N₂O reduction. Lowered pH in overlying water, enhanced adsorption of NH₄⁺-N and its nitrification rate likely contributed to the lower NH₃ volatilization as result of MBC addition. The nirS and nosZ gene copies were not changed by MBC, while the nirK gene copies were decreased as result of MBC amendment by 8.3%–25.2% under Hydragric Anthrosol and by 21.8%–24.9% under Haplic Acrisol. Consequent lower ratio of nirK/(nirS + nosZ) explained the mitigation effect of MBC on N₂O emission. In conclusion, the present two-year study recommends that MBC applied at a low dosage can perform positive effect on controlling the nitrogenous gas pollutants from paddy soil.

Arsenic (As) and cadmium (Cd), two major carcinogenic heavy metals, enters into human food chain by the consumption of rice or rice-based food products. Both As and Cd disturb plant-nutrient homeostasis and hence, reduces plant growth and crop productivity. In the present study, As/Cd modulated responses were studied in non-basmati (IR-64) and basmati (PB-1) rice varieties, at physiological, biochemical and transcriptional levels. At the seedling stage, PB-1 was found more sensitive than IR-64, in terms of root biomass; however, their shoot phenotype was comparable under As and Cd stress conditions. The ionomic data revealed significant nutrient deficiencies in As/Cd treated-roots. The principal component analysis identified NH₄⁺ as As-associated key macronutrient; while, NH₄⁺/NO₃⁻ and K⁺ was majorly associated with Cd mediated response, in both IR-64 and PB-1. Using a panel of 21 transporter gene expression, the extent of nutritional deficiency was ranked in the order of PB-1(As)<IR-64(As)<PB-1(Cd)<IR-64(Cd). A feed-forward model is proposed to explain nutrient deficiency induced de-regulation of gene expression, as observed under Cd-treated IR-64 plants, which was also validated at the level of sulphur metabolism related enzymes. Using urea supplementation, as nitrogen-fertilizer, significant mitigation was observed under As stress, as indicated by 1.018- and 0.794-fold increase in shoot biomass in IR-64 and PB-1, respectively compared to that of control. However, no significant amelioration was observed in response to supplementation of urea under Cd or potassium under As/Cd stress conditions. Thus, the study pinpointed the relative significance of various macronutrients in regulating As- and Cd-tolerance and will help in designing suitable strategies for mitigating As and/or Cd stress conditions.

Nitrogen (N) addition and mowing can significantly influence micronutrient cycling in grassland ecosystems. It remains largely unknown about how different forms of added N affect micronutrient status in plant-soil systems. We examined the effects of different N compounds of (NH₄)₂SO₄, NH₄NO₃, and urea with and without mowing on micronutrient Fe, Mn, Cu, and Zn in soil-plant systems in a meadow steppe. The results showed that (NH₄)₂SO₄ addition had a stronger negative effect on soil pH compared with NH₄NO₃ and urea, resulting in higher increases in soil available Fe and Mn herein. Nitrogen addition decreased plant community-level biomass weighted (hereafter referred to as community-level) Fe concentration but increased Mn concentration, with a greater effect under (NH₄)₂SO₄ addition. Community-level Cu concentration increased with (NH₄)₂SO₄ and NH₄NO₃ addition only under mowing treatment. Mowing synergistically interacted with urea addition to increase community-level Mn and Zn concentrations even with decreased soil organic matter, possibly because of compensatory plant growth and thus higher plant nutrient uptake intensity under mowing treatment. Overall, responses of plant-soil micronutrients to N addition varied with mowing and different N compounds, which were mainly regulated by soil physicochemical properties and plant growth. Different magnitude of micronutrient responses in plants and soils shed light on the necessity to consider the role of various N compounds in biogeochemical models when projecting the effects of N enrichment on grassland ecosystems.

It remains unclear how the source and rate of nitrogen (N) fertilizers affect N₂O concentration and effluxes along the soil profile under the drip-fertigated agricultural system. A plot-based field study was performed in 2017 and 2018 in a cotton field in arid northwestern China, with an objective to elucidate the impact of the applications of conventional urea (Urea), polymer-coated urea (ESN) and stabilized urea (SuperU) at rates of 120 and 240 kg N ha⁻¹ on concentration and efflux of N₂O in the soil profile and its relationship with N₂O surface emissions. The in-situ N₂O concentrations at soil depths of 5, 15, 30 and 60 cm were measured and used to estimate soil profile N₂O effluxes. Estimates of surface N₂O flux using the concentration gradient-based (GM) were compared with those measured using the chamber-based (CM) method. In both years, soil N₂O concentrations at all depths increased in response to basal N application at planting or in-season fertigation events. However, N rate or source did not affect soil N₂O concentrations or effluxes at each depth. Surface emissions of N₂O were mostly associated with that presented in the top layer of 0–15 cm. Surface N₂O efflux determined by GM was poorly or not associated with those of chamber measurements, which was attributed to the low N₂O production restricted by soil moisture condition under the drip-fertigated condition. These results highlight the challenge of applying the enhanced efficiency N fertilizer products in the drip-fertigated agricultural system.

Potato (Solanum tuberosum L.) production in irrigated coarse-textured soils requires intensive nitrogen (N) fertilization which may increase reactive N losses. Biological soil additives including N-fixing microbes (NFM) have been promoted as a means to increase crop N use efficiency, though few field studies have evaluated their effects, and none have examined the combined use of NFM with microbial inhibitors. A 2-year study (2018–19) in an irrigated loamy sand quantified the effects of the urease inhibitor NBPT, the nitrification inhibitor DMPSA, NFM, and the additive combinations DMPSA + NBPT and DMPSA + NFM on potato performance and growing season nitrous oxide (N₂O) emissions and nitrate (NO₃⁻) leaching. All treatments, except a zero-N control, received diammonium phosphate at 45 kg N ha⁻¹ and split applied urea at 280 kg N ha⁻¹. Compared with urea alone, DMPSA + NBPT reduced NO₃⁻ leaching and N₂O emissions by 25% and 62%, respectively, and increased crop N uptake by 19% in one year, although none of the additive treatments increased tuber yields. The DMPSA and DMPSA + NBPT treatments had greater soil ammonium concentration, and all DMPSA-containing treatments consistently reduced N₂O emissions, compared to urea-only. Use of NBPT by itself reduced NO₃⁻ leaching by 21% across growing seasons and N₂O emissions by 37% in 2018 relative to urea-only. In contrast to the inhibitors, NFM by itself increased N₂O by 23% in 2019; however, co-applying DMPSA with NFM reduced N₂O emissions by ≥ 50% compared to urea alone. These results demonstrate that DMPSA can mitigate N₂O emissions in potato production systems and that DMPSA + NBPT can reduce both N₂O and NO₃⁻ losses and increase the N supply for crop uptake. This is the first study to show that combining a nitrification inhibitor with NFM can result in decreased N₂O emissions in contrast to unintended increases in N₂O emissions that can occur when NFM is applied by itself.

Bio-stimulation of the indigenous microbial community is considered as an effective strategy for the bioremediation of polluted environments. This examination explored the near effects of various bio-stimulants on pyrene degradation, prokaryotic community compositions, and functions using 16S rRNA amplicon sequencing and qPCR. At first, the results displayed significant differences (p < 0.05) between the prokaryotic community structures of the control group, PYR (contains pyrene only), and bio-stimulants amended groups. Among the bio-stimulants, biochar, oxalic acid, salicylate, NPK, and ammonium sulfate augmented the pyrene degradation potential of microbial communities. Moreover, the higher abundance of genera, such as Flavobacterium, Hydrogenophaga, Mycobacterium, Rhodococcus, Flavihumibacter, Pseudomonas, Novosphingobium, etc., across the treatments indicated that these genera play a vital role in pyrene metabolism. Based on the higher abundance of GP-RHD and nidA genes, we speculated that Gram-positive prokaryotic communities are more competent in pyrene dissipation than Gram-negative. Furthermore, the marked abundance of nifH, and pqqC genes in the NPK and SA treatments, respectively, suggested that different bio-stimulants might enrich certain bacterial assemblages. Besides, the significant distinctions (p < 0.05) between the bacterial consortia of HA (humic acid) and SA (sodium acetate) groups from NPK, OX (oxalic acid), UR (urea), NH4, and SC (salicylate) groups also suggested that different bio-stimulants might induce distinct ecological impacts influencing the succession of prokaryotic communities in distinct directions. This work provides new insight into the bacterial degradation of pyrene using the bio-stimulation technique. It suggests that it is equally important to investigate the community structure and functions along with studying their impacts on degradation when devising a bio-stimulation technology.

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.

Herein, we report the optimization of nitrogen (N) doping in TiO₂ nanotubes to achieve the enhanced photocatalytic efficiencies in degradation of dye and H₂ gas evolution under solar light exposure. TiO₂ nanotubes have been produced via hydrothermal process and N doping has been tuned by varying the concentration of urea, being the source for N, by solid-state dispersion process. The structural analysis using XRD showed the characteristic occupancy of N into the structure of TiO₂ and the XPS studies showed the existence of Ti–N–Ti network in the N-doped TiO₂ nanotubes. The obtained TEM images showed the formation of 1D tube-like structure of TiO₂. Diffuse reflectance UV–Vis absorption spectra demonstrated that the N-doped TiO₂ nanotubes can efficiently absorb the photons of UV–Vis light of the solar light. The optimized N-doped TiO₂ nanotubes (TiO₂ nanotubes vs urea @ 1:1 ratio) showed the highest degradation efficiency over methyl orange dye (∼91% in 90 min) and showed the highest rate of H₂ evolution (∼19,848 μmol h⁻¹.g⁻¹) under solar light irradiation. Further, the recyclability studies indicated the excellent stability of the photocatalyst for the durable use in both the photocatalytic processes. The observed efficiency was ascribed to the optimized doping of N-atoms into the lattices of TiO₂, which enhanced the optical properties by forming new energy levels of N atoms near the valence band maximum of TiO₂, thereby increased the overall charge separation and recombination resistance in the system. The improved reusability of photocatalyst is attributed to the doping-induced structural stability in N-doped TiO₂. From the observed results, it has been recognized that the established strategy could be promising for synthesizing N-doped TiO₂ nanotubes with favorable structural, optical and photocatalytic properties towards dye degradation and hydrogen production applications.

The emergence of organophosphorus flame retardants and the efficient removal from aquatic environments have aroused increasing concerns. The Urea functionalized Fe₃O₄@LDH (Urea-Fe₃O₄@LDH) was prepared and used to adsorb triphenyl phosphate (tphp) for the first time. The tphp adsorption capacity was up to 589 mg g⁻¹, and the adsorption rate reached 49.9 mg g⁻¹ min⁻¹. Moreover, the influences of various environmental factors (pH, ionic strength and organic matter) on the tphp adsorption on the Urea-Fe₃O₄@LDH were investigated. The initial pH of the solution significantly affected the tphp adsorption, whereas the ionic strength and HA slightly affected the adsorption. The main adsorption mechanism was attributed to electrostatic interaction and π-π interaction. We believe that urea is one of excellent functional groups for the tphp adsorption removal and the materials with urea groups as the adsorbents exhibit good prospects in the future.