Reduced graphene oxide promotes the biodegradation of sulfamethoxazole by white-rot fungus Phanerochaete chrysosporium under cadmium stress.

The biodegradation of antibiotics in aquatic environment is consistently impeded by the widespread presence of heavy metals, necessitating urgent measures to mitigate or eliminate this environmental stress. This work investigated the degradation of sulfamethoxazole (SMX) by the white-rot fungus Phanerochaete chrysosporium (WRF) under heavy metal cadmium ion (Cd2+) stress, with a focus on the protective effects of reduced graphene oxide (RGO). The pseudo-first-order rate constant and removal efficiency of 5 mg/L SMX in 48 h by WRF decrease from 0.208 h-1 and 55.6% to 0.08 h-1 and 28.6% at 16 mg/L of Cd2+, while these values recover to 0.297 h-1 and 72.8% by supplementing RGO. The results demonstrate that RGO, possessing excellent biocompatibility, effectively safeguard the mycelial structure of WRF against Cd2+ stress and provide protection against oxidative damage to WRF. Simultaneously, the production of manganese peroxidase (MnP) by WRF decreases to 38.285 U/L in the presence of 24 mg/L Cd2+, whereas it recovers to 328.51 U/L upon the supplement of RGO. RGO can induce oxidative stress in WRF, thereby stimulating the secretion of laccase (Lac) and MnP to enhance the SMX degradation. The mechanism discovered in this study provides a new strategy to mitigate heavy metal stress encountered by WRF during antibiotic degradation.

Phytotoxicity of metal-organic framework MOF-74(Co) nanoparticles to pea seedlings.

Metal-organic framework (MOF) materials have unique structure and fantastic properties for wide-ranging applications. Pilot studies highlighted the toxicity and potential threats of MOF materials to the environment. In this study, we revealed the phytotoxicity of MOF-74(Co) nanoparticles (NPs) and their inhibitory effects on the photosynthesis of pea seedlings (Pisum sativum L.). MOF-74(Co) NPs have limited influences on the germination of pea seeds, but distinct environmental effects of MOF-74(Co) NPs were found in pea seedlings. The root length of pea seedlings, fresh weight and dry weight decreased by 50.0%, 29.2% and 36.4%, respectively, compared with the control group, when the material concentration was greater than 100 mg L-1. The net photosynthetic rate decreased by 48% and the intercellular CO2 concentration increased by 183% upon exposure to MOF-74(Co) NPs. Mechanistically, MOF-74(Co) exposure led to Co uptake in pea seedlings; the increases were 223% for the root, 267% for the stem and 6562% for the leaves, respectively, when the MOF-74(Co) NP concentration was 10 mg L-1. The released Co ions from MOF-74(Co) NPs caused oxidative damage to leaves and induced damage to the acceptor side of photosynthesis system II. Our results indicated that the environmental toxicity of MOF materials was largely regulated by the metal centers. MOF materials with nontoxic metal elements are desirable for future applications.

Toxicity of VO2 micro/nanoparticles to nitrogen-fixing bacterium Azotobacter vinelandii.

Vanadium dioxide (VO2) has been used in a variety of products due to its outstanding phase transition properties. However, as potential heavy metal contaminants, the environmental hazards and risks of VO2 should be systematically investigated. Biological nitrogen fixation is one of the most dominant processes in biogeochemical cycle, which is associated with nitrogen-fixing bacteria. In this study, we reported the environmental bio-effects of VO2 micro/nanoparticles on the nitrogen-fixing bacterium Azotobacter vinelandii. VO2 at 10 and 30 mg/L caused severe hazards to A. vinelandii, such as cell apoptosis, oxidative damage, physical damage, genotoxicity, and the loss of nitrogen fixation activity. The up-regulated differentially expressed genes of A. vinelandii were related to stress response, and the down-regulated genes were mainly related to energy metabolism. Surprisingly, VO2 of 10 mg/L decreased the nif gene expression but elevated the vnf gene expression, which enhanced the ability of A. vinelandii to reduce acetylene in anaerobic environment. In addition, under tested conditions, VO2 nanoparticles exhibited insignificantly higher toxicity than VO2 microparticles.

How accompanying ions affect soil heavy metal removal by polyepoxysuccinic acid during washing?

Soil washing with green eluent is an efficient technique to remediate heavy metal contaminated farmland. In addition to eluent, less is known about the roles of accompanying ions on heavy metal removal. We investigated the effects of accompanying ions including Fe3+, Mn2+, Ca2+, Mg2+, Al3+, Si4+ and PO43- on the desorption of Pb2+ and Cd2+ in paddy and arid soils using ethylenediaminetetraacetic acid and polyepoxysuccinic acid as eluents. The release rates of target and accompanying ions showed significant fast and slow reaction stages based on corresponding analysis and kinetic models. In fast reaction stage, Pb2+ and Cd2+ performed geochemical analogy with Ca2+, Mg2+ and PO43-. The release curves of these ions were fitted well with Elovich model, indicating that they released from oxysalt surface into solution via ion exchange, and dissolution of Fe/Mn/Al/Si (hydr)oxides through H+- and ligand-promoted dissolution. In slow reaction stage, Pb2+ and Cd2+ were related to Fe3+, Mn2+, Al3+ and Si4+, which were controlled by intraparticle diffusion process. H+ slowly diffused into interlayer of phyllosilicates to displace target and accompanying ions by ion exchange. Therefore, this research filled the gap of accompanying ions driving the release behavior of heavy metal ions during leaching.

Organic ligands regulate the environmental impacts of metal-organic frameworks on nitrogen-fixing bacterium Azotobacter vinelandii.

Metal-organic frameworks (MOFs) are rapidly developed materials with fantastic properties and wide applications. The increasing studies highlighted the potential threats of MOF materials to the environment. Comparing to the limited species of metal elements, the organic ligands have much higher diversity, but the influence of organic ligands on the environmental impacts of MOFs has not been revealed. Herein, we synthesized three Cu-MOFs with different organic ligands, namely Cu-BDC (1,4-terephthalic acid), Cu-IM (imidazole) and Cu-TATB (2,4,6-tris(4-carboxyphenyl)- 1,3,5-triazine), and evaluated their environmental toxicity to the nitrogen-fixing bacterium Azotobacter vinelandii. Cu-BDC inhibited the bacterial growth at lower concentrations than Cu-IM and Cu-TATB. The transcriptomes suggested the changes of membrane components by Cu-MOFs, consistent with the membrane leakage and cell wall damages. Cu-MOFs inhibited the nitrogen fixation activity through energy metabolism disturbance according to Gene Ontology functional annotation of ATP binding, Ca2+Mg2+-ATPase activity and ATP content. Only Cu-IM lowered the nitrogen fixation related nif genes, and affected the ribosome, purine metabolism and oxidative phosphorylation pathways. Otherwise, Cu-BDC and Cu-TATB mainly affected the flagellar assemblies and bacterial chemotaxis pathways. Our results collectively indicated that organic ligands regulated the environmental toxicity of MOFs through different metabolism pathways.

Particulate toxicity of metal-organic framework UiO-66 to white rot fungus Phanerochaete chrysosporium.

Metal-organic frameworks (MOF) are emerging materials with fantastic properties and wide applications. The release of metal ions from MOF materials is usually regarded as the origin of soluble MOF toxicity. However, whether the stable MOF particulates would induce environmental hazards is not clear. Herein, we aimed to reveal the particulate toxicity of MOF materials using the insoluble UiO-66 as the representative MOF and Phanerochaete chrysosporium as the model microorganism. UiO-66 nanoparticles (NPs) were synthesized by solvothermal method and their diameter was 68.4 ± 8.5 nm. UiO-66 NPs were stable in the culture system and the dissolution rate of 500 mg/L group was 0.26% after 14 d incubation. UiO-66 NPs did not affect the fungus growth according to the fresh weight increases and unchanged dry weights. Fungus mycelia kept even at concentrations up to 500 mg/L. Ultrastructural observation showed that UiO-66 NPs did not enter the fungal cells, but slightly destroyed the cell wall. UiO-66 NPs inhibited the laccase activity and promoted the activity of manganese peroxidase. The overall impact on the decomposition activity of P. chrysosporium was low in dye coloration test and sawdust degradation assay. Meaningful oxidative stress was aroused by UiO-66 NPs, as indicated by the decreases of catalase, glutathione, and total superoxide dismutase, and the increases of H2O2. Our results collectively suggested that the MOF particulates could induce mild mechanical damage to fungi and the toxicity was low comparing to other instable MOF materials.

Efficiency and comprehensive risk assessment of soil Pb and Cd by washing technique with three biodegradable eluents.

Soil washing with environmentally friendly eluents is a rapid remediation technique for farmland polluted by heavy metals. In this study, polyepoxysuccinic acid (PESA), ethylenediamine tetra (methylene phosphonic acid) sodium (EDTMPS), and phosphonyl carboxylic acid copolymer (POCA) were applied to remedy paddy and arid soils polluted by Pb and Cd. At the same time, ethylenediaminetetraacetic acid (EDTA) was used as a control eluent. PESA showed comparable removal of soil Pb and Cd (over 80.0%) with EDTA, and EDTMPS and POCA removed two heavy metals by 35.2-50.3%. For labile fractions, PESA significantly removed Pb by 93.5-96.7% and Cd by 84.9-90.3% in two soils. EDTMPS and POCA removed Pb by 75.5-85.8% in two soils, while they only removed Cd by 11.7-42.2% in paddy soil, and 76.3-81.7% in arid soil. The risks of total heavy metal concentrations were reduced from the high risk to low risk in paddy soil, and to considerable risk in arid soil, while only dropped to considerable or even had no change by EDTMPS and POCA leaching. The risks of the two soils reduced from high to low or considerable level after PESA washing based on labile fraction change, and to considerable or high level after EDTMPS and POCA leaching, respectively. Therefore, PESA is an ecological benefit eluent for remediating the farmland polluted by heavy metals, and the risk assessment based on labile fraction more easily identifies the dynamic change of heavy metal during the washing process.

Can heavy metal pollution induce bacterial resistance to heavy metals and antibiotics in soils from an ancient land-mine?

Microbial resistance to antibiotics is a growing challenge to human health. Recent evidence has indicated that antibiotic resistance can be co-selected for by exposure to heavy metals in agricultural soils. It remains unknown if this is a concern in other environments contaminated by metals. We here investigated soil microbial activities, composition and tolerance to heavy metals and antibiotics in a mining soil survey. We found that microbial respiration, growth, and biomass were affected by available metal concentrations. Most of the variation in microbial PLFA composition was explained by differences in heavy metal and pH. Additionally, pollution-induced bacterial community tolerance to toxicants including Cu, Pb, Zn, tetracycline and vancomycin was determined. Although only bacterial tolerance to Pb increased with higher levels of metals, the links between bacterial metal tolerance and soil metal concentrations were clear when considered together with previously published reports, suggesting that bacterial metal tolerance were universally elevated in the surveyed soils. The induced levels of heavy metal tolerance coincided with elevated levels of tolerance to vancomycin, but not to tetracycline. Our study showed that heavy metals can co-select for resistance to clinically important antibiotics also in ecosystems without manure input or antibiotic pollution.

Remediation of heavy metal contaminated soil by biodegradable chelator-induced washing: Efficiencies and mechanisms.

Biodegradable chelators (BCs) are promising substitutes for conventional washing agents in the remediation of heavy metal contaminated soil with strong complexing ability and less cost. However, great challenges for the applications of BC-assisted washing still exist, such as the assessment of the factor affecting the efficiency of metal removal and the unclear of the metal removal mechanism. Batch washing was therefore explored to evaluate the potential for four BCs for removing Cd, Pb, and Zn from polluted soils. The soil spectroscopic characteristics before and after washing were also investigated. The results demonstrated that iminodisuccinic acid (ISA) and glutamate-N, N-diacetic acid (GLDA) were an appealing alternative to commonly used non-biodegradable ethylenediaminetetraacetic acid, but glucomonocarbonic acid (GCA) and polyaspartic acid (PASP) were less efficient. Optimal parameters of BCs were determined to be a concentration of 50 mmol L-1, a pH of 5.0, a contact time of 120 min, and a solid/liquid ratio of 1:5, considering metal removal efficiencies and the suitable cost. A single removal washing could be up to 52.39% of Cd, 71.79% of Pb, and 34.13% of Zn from mine soil, and 98.28% of Cd, 91.10% of Pb, and 90.91% of Zn from polluted farmland soil. After washing, the intensity of heavy metal binding to soil colloids increased while the metal mobility reduced because of weakly bound fractions removed by BCs. The BCs-induced soil washing revealed that the possible mechanisms of metal removal included the acid dissolution, ion exchange, and surface complexation. Our findings highlight the potential application of especially ISA and GLDA as efficient washing agents to remove potentially toxic elements from contaminated soils.

Synchronous removal of ammonium and phosphate from swine wastewater by two agricultural waste based adsorbents: Performance and mechanisms.

Two agricultural wastes, Chinese medicinal herbal residue and spent Pleurotus ostreatus substrate, were developed to remove ammonium and phosphate from swine wastewater. These adsorbents were mesoporous materials with abundant smooth layered pores, and rough protuberances and grooves, respectively. Their adsorption capacities were 1131.65 and 1631.79 mg N g-1, and 63.41 and 62.58 mg P g-1 at pH 8.0, dosage of 0.2 g L-1 and contact time of 360 min. And kinetics data of ammonium and phosphate fitted best with the intra-particle diffusion and pseudo-second-order models, respectively. Based on the point of zero charge, FTIR and XPS analyses, ammonium was removed mainly by electrostatic attraction, ion exchange and surface precipitation, while phosphate was by ligand exchange, surface complexation and precipitation. Therefore, the two agricultural wastes have great potential to synchronously remove ammonium and phosphate from swine wastewater.

Effects of soil chemical properties and fractions of Pb, Cd, and Zn on bacterial and fungal communities.

Heavy metal contamination in soils poses a serious threat to microorganisms, which play important roles in soil biogeochemical process. However, the key fractions of heavy metals affecting soil microorganisms are still unclear. In this study, DNA sequencing, redundancy and variance partition analysis were performed to investigate the combined effects of heavy metal fractions and soil chemical properties on microbial communities in Pb, Cd, and Zn co-contaminated soils. The results showed that long-term exposure of microorganisms to these metals changed the richness, diversity, and structure of their communities. The bacterial and fungal Chao richness indexes decreased, but only the bacterial Shannon index improved with increasing metal concentrations. Moreover, soil available potassium and acid-extractable Pb made the greatest contributions to variations in the bacterial community structure, while soil pH, water-extractable Pb and Zn were the dominant factors influencing the fungal community structure. In addition, Marmoricola, Nocardioides, and Gibberella were sensitive to these metals. Overall, the effects of different heavy metal fractions on microorganisms varied significantly, and these metal fractions together with soil chemical properties determined the soil microbial communities.

Soil washing remediation of heavy metal from contaminated soil with EDTMP and PAA: Properties, optimization, and risk assessment.

Soil washing is a rapid and efficient remediation technique for soil contaminated by heavy metals. In this study, Cd, Pb, and Zn were removed from contaminated soil by ethylenediamine tetra (methylene phosphonic acid) (EDTMP) and polyacrylic acid (PAA). We then investigated the effect of varying the concentration, pH and duration of the washing processes. Single-factor experiments suggest that the PAA washing process may be dominated by electrostatic adsorption, and is suitable for remediation under weak acid and neutral conditions. Meanwhile, EDTMP remediation might be dominated by chelation, which is favorable in strong acid and alkaline environments. In a quadratic saturation D-optimization design (QSDD), we optimized the washing parameters and further explored the washing mechanism including primary factor, principal effect, interaction effect, and the optimal washing conditions, with simultaneously changing multiple influencing factors. The optimum efficiencies of Cd, Pb, and Zn removal were 92.74%, 96.14%, and 50.76% respectively in EDTMP remediation, and 84.62, 79.24, and 41.66% respectively in PAA remediation. The washing processes effectively reduced the availability of Cd, Pb, and Zn in contaminated soil, without noticeably affecting soil chemical properties. Therefore, the washing incurred little ecological risk. EDTMP and PAA are suitable remediation agents of soil contaminated by heavy metals.

Removal of lead, zinc and cadmium from contaminated soils with two plant extracts: Mechanism and potential risks.

Screening appropriate washing agents to remediate soils contaminated with heavy metals is crucial for decreasing metal hazards posing to environment and human health. In this study, two plant washing agents-water-extracted from Fagopyrum esculentum and Fordiophyton faberi, were applied to remove soil Pb, Zn, and Cd by washing. Results indicated that metal removals augmented with increase of washing solution concentrations, decreased with increasing pH values of the solution and followed the pseudo-second-order model depending on contact duration. At concentration of 50 g/L, pH 3 and contact duration of 120 min, F. esculentum had higher removals of Pb (5.98-6.83%), Zn (21.82-27.94%), and Cd (39.90-40.74%) than those of F. faberi. And metal ions could be removed by binding with carboxyl, hydroxyl, amide, amine and aromatic groups in washing solutions. The potential risks of residual metals declined by 51.35-52.12% for mine soil and 48.51-49.96% for farmland soil with exchangeable and carbonate-bound fractions obviously extracted after a single washing (P < 0.05). And soil organic carbon and nutrients increased to some extent except for total phosphorus and available potassium. Moreover, soil phytotoxicity lowered except that some adverse effects on seed germination existed. Therefore, the water extract from F. esculentum is a promising washing agent for heavy metal removal.

Effect of soil washing with biodegradable chelators on the toxicity of residual metals and soil biological properties.

Soil washing with chelators is a promising and efficient method of remediating metals-contaminated soils. However, the toxicity of residual metals and the effects on soil microbial properties have remained largely unknown after washing. In this study, we employed four biodegradable chelators for removal of metals from contaminated soils: iminodisuccinic acid (ISA), glutamate-N,N-diacetic acid (GLDA), glucomonocarbonic acid (GCA), and polyaspartic acid (PASP). The maximum removal efficiencies for Cd, Pb, and Zn of 85, 55, and 64% and 45, 53, and 32% were achieved from farmland soil and mine soil using biodegradable chelators, respectively. It was found that the capacity of ISA and GLDA to reduce the labile fraction of Cd, Pb, and Zn was similar to that of the conventional non-biodegradable chelator ethylenediaminetetraacetic acid (EDTA). The leachability, mobility, and bioaccessibility of residual metals after washing decreased notably in comparison to the original soils, thus mitigating the estimated environmental and human health risks. Soil ß-glucosidase activity, urease activity, acid phosphatase activity, microbial biomass nitrogen, and microbial biomass phosphorus decreased in the treated soils. However, compared with EDTA treatment, soil enzyme activities distinctly increased by 5-94% and overall microbial biomass slightly improved in the remediated soils, which would facilitate reuse of the washed soils. Based on soil toxicity tests that employed wheat seed germination as the endpoint of assessment, the washed soils exhibited only slight effects especially after ISA and GLDA treatments, following high-efficiency metal removal. Hence, ISA and GLDA appear to possess the greatest potential to rehabilitate polluted soils with limited toxicity remaining.

Feasibility of nanoscale zero-valent iron to enhance the removal efficiencies of heavy metals from polluted soils by organic acids.

Soil washing with natural chelators to remediate metal-contaminated soils has been gained attention by researchers. However, the abilities of the chelators to remediate the multiple metal polluted soils are less effective. This study employed zero-valent iron nanoparticle (nZVI) to enhance the removal efficiencies of citric (CA), tartaric (TA) and oxalic acids (OA), and evaluate their feasibility. Results showed that metal removal efficiencies increased with the increasing concentration of nZVI and soil-liquid ratio, decreased with the increasing solution pH. The kinetic simulation indicated that pseudo-first-order and pseudo-second-order models could be used for describing the washing processes. Additionally, metal removals were significantly improved by addition of nZVI (p < 0.05). The highest enhancements of soil Cd, Pb and Zn removals under solution pH of 4.0, soil-liquid ratio of 1:20 and washing time of 120 min reached 12.83% (OA- nZVI), 24.92% (CA-nZVI) and 11.64% (OA- nZVI) for mine soil, and 19.24% (TA- nZVI), 18.16% (CA-nZVI) and 8.93% (OA- nZVI) for farmland soil, respectively. After soil washing, the exchangeable forms and the environmental risks of residual metals were markedly diminished in soils. Therefore, the combinations of the organic acids and nZVI are the feasible practices to repair the soils contaminated by heavy metals.

Feasibility of four wastes to remove heavy metals from contaminated soils.

Soil washing is one of the permanent techniques to remove heavy metals, and washing agent is a key influence factor for this technique, but there is still lack of high-efficiency, eco-friendly, and inexpensive agents. In this study, four wastes including pineapple peel (PP), soybean straw (SS), broad bean straw (BBS) and tea residue (TR) were employed to remove cadmium (Cd), lead (Pb) and zinc (Zn) in contaminated soils. The Fourier transform infrared spectroscopy (FTIR) analysis indicated that hydroxyl, carboxyl, amine, carbonyl and amide groups were involved in the interaction with metal ions by complexation or ion exchange. We then investigated the influences of various conditions including washing solution concentration, pH, and washing time. The metal removal efficiencies with these agents increased as the concentration augmented from 5 to 80 g L-1, decreased or presented an asymmetric V-shaped curve with increasing pH from 2.5 to 7.5, and fit intraparticle diffusion or Elovich model with washing time increasing. PP has the highest removals for Cd (90.1%), Pb (18.6%), and Zn (15.2%) in soil A, and 85.8, 24.8, and 69.4% in soil B, respectively. The relatively high metal removal was mainly attributed to effective removal of the exchangeable and acid soluble fractions. Moreover, single washing not only lowered the potential ecological risk of the heavy metals, but moderated the effects on soil chemical properties. Therefore, PP was a feasible washing agent to remediate soils contaminated by heavy metals.

Heavy metals in soils from a typical industrial area in Sichuan, China: spatial distribution, source identification, and ecological risk assessment.

Anthropogenic activities could result in increasing concentrations of heavy metals in soil and deteriorating in soil environmental quality. Topsoil samples from a typical industrial area, Shiting River Valley, Sichuan, Southwest China, were collected and determined for the concentrations of Cu, Zn, Cr, Cd, As, and Hg. The mean concentrations of these metals were lower than the national threshold values, but were slightly higher than their corresponding background values, indicating enrichment of these metals in soils in the valley, especially for Cu, Zn, and Hg. The topsoils in this area demonstrated moderate pollution and low potential ecological risk. Principal component analysis coupled with cluster analysis was applied to analyze the data and identified possible sources of these heavy metals; the results showed that soil Cd, Hg, As, Cu, and Zn were predominantly controlled by human activities, whereas Cr was mainly from the parent material. The spatial distribution of the heavy metals varied distinctly and was closely correlated to local anthropogenic activities. Furthermore, the concentrations of heavy metals in the industrial land demonstrated relatively higher levels than those of other land use patterns. Soil metal concentrations decreased with the distance increase from the traffic highway (0-1.0 km) and water system (0-2.0 km). Additionally, soil properties, especially pH and soil organic matter, were found to be important factors in the distribution and composition of metals.

Heavy metal removal by GLDA washing: Optimization, redistribution, recycling, and changes in soil fertility.

Soil washing, an emerging method for treating soils contaminated by heavy metals, requires an evaluation of its efficiency in simultaneously removing different metals, the quality of the soil following remediation, and the reusability of the recycled washing agent. In this study, we employed N,N-bis (carboxymethyl)-l-glutamic acid (GLDA), a novel and readily biodegradable chelator to remove Cd, Pb, and Zn from polluted soils. We investigated the influence of washing conditions, including GLDA concentration, pH, and contact time on their removal efficiencies. The single factor experiments showed that Cd, Pb, and Zn removal efficiencies reached 70.62, 74.45, and 34.43% in mine soil at a GLDA concentration of 75mM, a pH of 4.0, and a contact time of 60min, and in polluted farmland soil, removal efficiencies were 69.12, 78.30, and 39.50%, respectively. We then employed response surface methodology to optimize the washing parameters. The optimization process showed that the removal efficiencies were 69.50, 88.09, and 40.45% in mine soil and 71.34, 81.02, and 50.95% in polluted farmland soil for Cd, Pb, and Zn, respectively. Moreover, the overall highly effective removal of Cd and Pb was connected mainly to their highly effective removal from the water-soluble, exchangeable, and carbonate fractions. GLDA-washing eliminated the same amount of metals as EDTA-washing, while simultaneously retaining most of the soil nutrients. Removal efficiencies of recycled GLDA were no >5% lower than those of the fresh GLDA. Therefore, GLDA could potentially be used for the rehabilitation of soil contaminated by heavy metals.

Spatial variability of soil nitrogen in a hilly valley: Multiscale patterns and affecting factors.

Estimating the spatial distribution of soil nitrogen at different scales is crucial for improving soil nitrogen use efficiency and controlling nitrogen pollution. We evaluated the spatial variability of soil total nitrogen (TN) and available nitrogen (AN) in the Fujiang River Valley, a typical hilly region composed of low, medium and high hills in the central Sichuan Basin, China. We considered the two N forms at single hill, landscape and valley scales using a combined method of classical statistics, geostatistics and a geographic information system. The spatial patterns and grading areas of soil TN and AN were different among hill types and different scales. The percentages of higher grades of the two nitrogen forms decreased from low, medium to high hills. Hill type was a major factor determining the spatial variability of the two nitrogen forms across multiple scales in the valley. The main effects of general linear models indicated that the key affecting factors of soil TN and AN were hill type and fertilization at the single hill scale, hill type and soil type at the landscape scale, and hill type, slope position, parent material, soil type, land use and fertilization at the valley scale. Thus, the effects of these key factors on the two soil nitrogen forms became more significant with upscaling.