Geochemical fractionation and potential release behaviour of heavy metals in lead‒zinc smelting soils.

The lack of understanding of heavy metal speciation and solubility control mechanisms in smelting soils limits the effective pollution control. In this study smelting soils were investigated by an advanced mineralogical analysis (AMICS), leaching tests and thermodynamic modelling. The aims were to identify the partitioning and release behaviour of Pb, Zn, Cd and As. The integration of multiple techniques was necessary and displayed coherent results. In addition to the residual fraction, Pb and Zn were predominantly associated with reducible fractions, and As primarily existed as the crystalline iron oxide-bound fractions. AMICS quantitative analysis further confirmed that Fe oxyhydroxides were the common dominant phase for As, Cd, Pb and Zn. In addition, a metal arsenate (paulmooreite) was an important mineral host for Pb and As. The pH-stat leaching indicted that the release of Pb, Zn and Cd increased towards low pH values while release of As increased towards high pH values. The separate leaching schemes were associated with the geochemical behaviour under the control of minerals and were confirmed by thermodynamic modelling. PHREEQC calculations suggested that the formation of arsenate minerals (schultenite, mimetite and koritnigite) and the binding to Fe oxyhydroxides synchronously controlled the release of Pb, Zn, Cd and As. Our results emphasized the governing role of Fe oxyhydroxides and secondary insoluble minerals in natural attenuation of heavy metals, which provides a novelty strategy for the stabilization of multi-metals in smelting sites.

Microbial community composition and cooccurrence patterns driven by co-contamination of arsenic and antimony in antimony-mining area.

In the current study, a typical Sb mine was selected to explore the microbial community composition and assembly driven by the cocontamination of As/Sb with geographic distance. Our results showed that environmental parameters, especially pH, TOC, nitrate, total and bioavailable As/Sb contents largely affected the microbial community diversity and composition. The total and bioavailable As/Sb levels were significantly positively correlated with the relative abundance of Zavarzinella, Thermosporothrix and Holophaga, while the pH presented a significant negative correlation with the three genera, potentially implying they are important taxonomic groups in acid-mining soils. The cooccurrence network analysis indicated the environmental stress dominated by pH and As/Sb co-contamination affected the microbial modularity and interaction. Meanwhile, Homogeneous selection (HoS, 26.4-49.3%), and drift and others (DR, 27.1∼40.2%) were the most important assembly processes for soil bacterial, and the importance of HoS decreased and the DR increased with geographic distance to the contamination source respectively. Soil pH, nutrient availability, total and bioavailable As/Sb contents significantly affected the HoS and DR processes. This study provides theoretical support for microbial remediation in metal(loid)-contaminated soils.

The effect of extracellular polymeric substances (EPS) of iron-oxidizing bacteria (Ochrobactrum EEELCW01) on mineral transformation and arsenic (As) fate.

Extracellular polymeric substances (EPS) are an important medium for communication and material exchange between iron-oxidizing bacteria and the external environment and could induce the iron (oxyhydr) oxides production which reduced arsenic (As) availability. The main component of EPS secreted by iron-oxidizing bacteria (Ochrobactrum EEELCW01) was composed of polysaccharides (150.76-165.33 mg/g DW) followed by considerably smaller amounts of proteins (12.98-16.12 mg/g DW). Low concentrations of As (100 or 500 µmol/L) promoted the amount of EPS secretion. FTIR results showed that EPS was composed of polysaccharides, proteins, and a miniscule amount of nucleic acids. The functional groups including -COOH, -OH, -NH, -C=O, and -C-O played an important role in the adsorption of As. XPS results showed that As was bound to EPS in the form of As3+. With increasing As concentration, the proportion of As3+ adsorbed on EPS increased. Ferrihydrite with a weak crystalline state was only produced in the system at 6 hr during the mineralization process of Ochrobactrum sp. At day 8, the minerals were composed of goethite, galena, and siderite. With the increasing mineralization time, the main mineral phases were transformed from weakly crystalline hydrous iron ore into higher crystallinity siderite (FeCO3) or goethite (α-FeOOH), and the specific surface area and active sites of minerals were reduced. It can be seen from the distribution of As elements that As is preferentially adsorbed on the edges of iron minerals. This study is potential to understand the biomineralization mechanism of iron-oxidizing bacteria and As remediation in the environment.

Arsenic Elevated Groundwater Irrigation: Farmers' Perception of Rice and Vegetable Contamination in a Naturally Arsenic Endemic Area.

Arsenic (As) in groundwater and its accumulation in agricultural produces has caused serious threats to human health. The majority of current research on As mainly focuses on the technical aspects while bypassing the social perspectives. Farmers are the prime stakeholders as well as executors of agricultural strategies, and their adaptation largely depends on how they perceive the risk for which a mitigation strategy is proposed. This study aims to explore how rice and vegetable farmers perceive As accumulation in their rice and vegetables as well as explore current crop- and body-loading status, the subsequent health consequences of As, and alleviation possibilities with mitigation strategies and to investigate if there is an association between their socioeconomic status and their level of perception. Results reveal that one-fourth of the farmers gave a positive message regarding the As-contamination scenario in rice and vegetables. Although 10 farmers' socioeconomic characteristics were positively significant, distinctive emphasis should be given to five predictor variables explaining 88% variances: knowledge, direct participation in farming, information sources used, participant education, and organizational participation. Path analysis depicts that direct participation in farming presents the highest positive total effect (0.855) and direct effect (0.503), whereas information sources show the highest positive indirect effect (0.624). The mean As content in all five locations was statistically significant at the 5%, 5%, 0.1%, 1%, and 1% probability levels in scalp hairs, rice, vegetables, soils, and irrigation water, respectively. The first principal component (PC1) explains 92.5% of the variation. Significant variations were primarily explained by As levels in irrigation water, rice grain, and soil. Farmers' perception is far behind the actual field status of As level and its transfer. Therefore, intensified priorities should be administered on the farmers' characteristics contributing to variances in perception. The findings can be utilized for policy formulation in all As-endemic nations. More multidisciplinary research can be undertaken on farmers' attitude towards adopting As-mitigation techniques, with a focus on the socioeconomic position found to influence farmers' perceptions.

Light plays a critical role in the accumulation of chlorogenic acid in Lonicera macranthoides Hand.-Mazz.

Light has important effects on plant metabolism. However, the relationship between the chlorogenic acid (CGA) content and light in plants remains unclear. Here, we investigated the effects of shading treatment on gene expression and CGA content in Lonicera macranthoides Hand.-Mazz. (LM), a widely used medicinal plant. A total of 1891 differentially expressed genes (DEGs) were obtained in flower buds and 819 in leaves in response to light in shading treatment compared to the control sample by RNA-Seq. After shading treatment, the content of CGA in LM leaves decreased significantly by 1.78-fold, the carotenoid content increased, and the soluble sugar and starch contents significantly decreased. WGCNA and the expression of related genes verified by qRT‒PCR revealed that CGA synthesis pathway enzyme genes form a co-expression network with genes for carbohydrate synthesis, photosynthesis, light signalling elements, and transcription factor genes (TFs) that affect the accumulation of CGA. Through a virus-induced gene silencing (VIGS) system and CGA assay in Nicotiana benthamiana (NB), we determined that downregulation of NbHY5 expression decreased the CGA content in NB leaves. In this study, we found that light provides energy and material for the accumulation of CGA in LM, and light affects the expression of CGA accumulation-related genes. Our results show that different light intensities have multiple effects on leaves and flower buds in LM and are able to coregulate LmHY5 expression and CGA synthesis.

Soil heavy metal pollution from Pb/Zn smelting regions in China and the remediation potential of biomineralization.

Smelting activities pose serious environmental problems due to the local and regional heavy metal pollution in soils they cause. It is therefore important to understand the pollution situation and its source in the contaminated soils. In this paper, data on heavy metal pollution in soils resulting from Pb/Zn smelting (published in the last 10 years) in China was summarized. The heavy metal pollution was analyzed from a macroscopic point of view. The results indicated that Pb, Zn, As and Cd were common contaminants that were present in soils with extremely high concentrations. Because of the extreme carcinogenicity, genotoxicity and neurotoxicity that heavy metals pose, remediation of the soils contaminated by smelting is urgently required. The primary anthropogenic activities contributing to soil pollution in smelting areas and the progressive development of accurate source identification were performed. Due to the advantages of biominerals, the potential of biomineralization for heavy metal contaminated soils was introduced. Furthermore, the prospects of geochemical fraction analysis, combined source identification methods as well as several optimization methods for biomineralization are presented, to provide a reference for pollution investigation and remediation in smelting contaminated soils in the future.

Arsenic biotransformation genes and As transportation in soil-rice system affected by iron-oxidizing strain (Ochrobactrum sp.).

Arsenic (As) biotransformation in soil affects As biogeochemical cycling and is associated with As accumulation in rice. After inoculation with 1% iron-oxidizing bacteria (FeOB) in paddy soil, As speciation, As biotransformation genes in soil, As/Fe in Fe plaques, and As accumulation in rice were characterized. Compared with the control, the available As concentrations in soils decreased while amorphous and poorly crystalline Fe-Al oxidized As and crystalline Fe-Al oxidized As fractions increased of F (FeOB) and RF (rice and FeOB) treatments. Fe concentrations increased and positively correlated with As concentrations in Fe plaques on the rice root surface (***P < 0.001). Compared with R (rice), Monomethyl As (MMA), dimethyl As (DMA), arsenate (As(V)), and arsenite (As(III)) concentrations in rice plants showed a downwards trend of RF treatment. The As concentration in grains was below the National Standard for Food Safety (GB 2762-2017). A total of 16 As biotransformation genes in rhizosphere soils of different treatments (CK, F, R and RF were quantified by high-throughput qPCR (HT-qPCR). Compared with the control, the As(V) reduction and As transport genes abundance in other treatments increased respectively by 54.54%-69.17% and 54.63%-73.71%; the As(III) oxidation and As (de) methylation genes did not change significantly; however, several As(III) oxidation genes (aoxA, aoxB, aoxS, and arsH) increased. These results revealed that FeOB could reduce, transport As, and maybe also oxidize As. In addition, As(III) oxidation gene (aoxC) in rhizosphere soil was more abundant than in non-rhizosphere soil. It indicated that radial oxygen loss (ROL) promoted As(III) oxidation in rhizosphere soils. The results provide evidence for As biotransformation by ROL and FeOB in soil-rice system. ROL affects As oxidation and immobilization, and FeOB affects As reduction, transportation and may also affect As oxidation.

Arsenic Transformation in Soil-Rice System Affected by Iron-Oxidizing Strain (Ochrobactrum sp.) and Related Soil Metabolomics Analysis.

Iron-oxidizing bacteria (FeOB) could oxidize Fe(II) and mediate biomineralization, which provides the possibility for its potential application in arsenic (As) remediation. In the present study, a strain named Ochrobactrum EEELCW01 isolated previously, was inoculated into paddy soils to investigate the effect of FeOB inoculation on the As migration and transformation in paddy soils. The results showed that inoculation of Ochrobactrum sp. increased the proportion of As in iron-aluminum oxide binding fraction, which reduced the As bioavailability in paddy soils and effectively reduced the As accumulation in rice tissues. Moreover, the inoculation of iron oxidizing bacteria increased the abundance of KD4-96, Pedosphaeraceae and other bacteria in the soils, which could reduce the As toxicity in the soil through biotransformation. The abundance of metabolites such as carnosine, MG (0:0/14:0/0:0) and pantetheine 4'-phosphate increased in rhizosphere soils inoculated with FeOB, which indicated that the defense ability of soil-microorganism-plant system against peroxidation caused by As was enhanced. This study proved that FeOB have the potential application in remediation of As pollution in paddy soil, FeOB promotes the formation of iron oxide in paddy soil, and then adsorbed and coprecipitated with arsenic. On the other hand, the inoculation of Ochrobactrum sp. change soil microbial community structure and soil metabolism, increase the abundance of FeOB in soil, promote the biotransformation process of As in soil, and enhance the resistance of soil to peroxide pollution (As pollution).

Remediation of arsenic-contaminated paddy field by a new iron oxidizing strain (Ochrobactrum sp.) and iron-modified biochar.

Iron-oxidizing strain (FeOB) and iron modified biochars have been shown arsenic (As) remediation ability in the environment. However, due to the complicated soil environment, few field experiment has been conducted. The study was conducted to investigate the potential of iron modified biochar (BC-FeOS) and biomineralization by a new found FeOB to remediate As-contaminated paddy field. Compared with the control, the As contents of GB (BC-FeOS), GF (FeOB), GFN (FeOB and nitrogen fertilizer), GBF (BC-FeOS and FeOB) and GBFN (BC-FeOS, FeOB and nitrogen fertilizer) treatments in pore water decreased by 36.53%-80.03% and the microbial richness of iron-oxidizing bacteria in these treatments increased in soils at the rice maturation stage. The concentrations of available As of GB, GF, GFN, GBF and GBFN at the tillering stage were significantly decreased by 10.78%-55.48%. The concentrations of nonspecifically absorbed and specifically absorbed As fractions of GB, GF, GFN, GBF and GBFN in soils were decreased and the amorphous and poorly crystalline hydrated Fe and Al oxide-bound fraction was increased. Moreover, the As contents of GB, GF, GFN, GBF and GBFN in rice grains were significantly decreased (*P < 0.05) and the total As contents of GFN, GBF and GBFN were lower than the standard limit of the National Standard for Food Safety (GB 2762-2017). Compared with the other treatments, GBFN showed the greatest potential for the effective remediation of As-contaminated paddy fields.

Effects of biochar/AQDS on As(III)-adsorbed ferrihydrite reduction and arsenic (As) and iron (Fe) transformation: Abiotic and biological conditions.

Microbe induced iron (Fe) reduction play an important role in arsenic (As) transformation and the related secondary mineral formation. Meanwhile biochar could react as electron shuttle for this process. Impact of biochar and model electron shuttle anthraquinone-2,6-disulfonate (AQDS) on the chemical/biological iron reduction of As(III)-adsorbed ferrihydrite and the solid-liquid redistribution of As in M1 buffer were studied. Fe reduction results in the release of As adsorbed on ferrihydrite into the solution. Under abiogenic conditions, both biochar and AQDS promoted ferrous production, the chemical oxidation of As(III) and As release. Inoculate with Shewanella oneidensis MR-1, AQDS has greater electronic shuttle function than biochar (with the maximum Fe(II) contents: 154 mg/L > 76.6 mg/L respectively). However, only 12.8 mg/L As was released in the presence of AQDS, which was much lower than that in the presence of biochar (21.6 mg/L), and may be associated with the transformation of As speciation and the formation of secondary minerals. XRD and EDX-SEM confirmed that the As could be fixed by the generated secondary mineral vivianite. The relative contents of vivianite in biological control and AQDS addition were 2.7% and 18.4%, respectively. This study provides information on the transformation and migration of As and Fe with the addition of biochar under anaerobic conditions, which is potential to understand the mechanism of As(III)-contaminated soil remediation.

Identification of reliable reference genes for quantitative real-time PCR analysis of the Rhus chinensis Mill. leaf response to temperature changes.

Rhus chinensis Mill. (RCM) is the host plant of Galla chinensis, which is valued in traditional medicine. Environmental temperature directly determines the probability of gallnut formation and RCM growth. At present, there is no experiment to systematically analyse the stability of internal reference gene (RG) expression in RCM. In this experiment, leaves that did not form gallnuts were used as the control group, while leaves that formed gallnuts were used as the experimental group. First, we conducted transcriptome experiments on RCM leaves to obtain 45 103 differential genes and functional enrichment annotations between the two groups. On this basis, this experiment established a transcriptional gene change model of leaves in the process of gallnut formation after being bitten by aphids, and RCM reference candidate genes were screened from RNA sequencing (RNA-seq) data. This study is based on RCM transcriptome data and evaluates the stability of 11 potential reference genes under cold stress (4 °C) and heat stress (34 °C), using three statistical algorithms (geNorm, NormFinder, and BestKeeper). The results show that GAPDH1 + PP2A2/UBQ are stable reference genes under heat stress, while GAPDH1 + ACT are the most stable under cold stress. This study is the first to screen candidate reference genes in RCM and could help guide future molecular studies in this genus.

Microbial driven iron reduction affects arsenic transformation and transportation in soil-rice system.

The microbe-driven iron cycle plays an important role in speciation transformation and migration of arsenic (As) in soil-rice systems. In this study, pot experiments were used to investigate the effect of bacterial iron (Fe) reduction processes in soils on As speciation and migration, as well as on As uptake in soil-rice system. During the rice growth period, pH and electrical conductivity (EC) in soil solutions initially increased and then decreased, with the ranges of 7.4-8.8 and 116.3-820 mS cm-1, respectively. The concentrations of Fe, total As and As(III) showed an increasing trend in the rhizosphere and non-rhizosphere soil solutions with the increasing time. Fe concentrations were significantly positively correlated with total As and As(III) concentrations (***p < 0.001) in the soil solutions. The abundances of the arsenate reductase gene (arsC) and the As(III) S-adenosylmethionine methyltransferase gene (arsM) in rhizosphere soils were higher than those in non-rhizosphere soils, while the abundance of the Fe-reducing bacteria (Geo) showed an opposite trend. Moreover, it showed that the Geo abundance was significantly positively correlated with that of the arsC (***p < 0.001) and arsM (**p < 0.01) genes, respectively. The abundances of Geo, arsC and arsM genes were significantly positively correlated with the concentrations of Fe, total As and As(III) in the soil solutions (*p < 0.05). Moreover, the abundances of arsC and arsM genes were significantly negatively correlated with total As and As(III) in rice grains (*P < 0.05). These results showed that the interaction of bacterial Fe reduction process and radial oxygen loss from roots promoted the reduction and methylation of As, and then decreased As uptake by rice, which provided a theoretical basis for alleviating As pollution in paddy soils.

Planting a Seed of Experience - Long Term Effects of a Co-curricular Ecogarden-Based Program in Higher Education in Hong Kong.

This paper reports on the long-term effectiveness of a non-formal co-curricular educational program based on a campus ecogarden at a Hong Kong university in developing pro-sustainability awareness, attitudes and behavior among undergraduate students. This service-based, nature-based experiential learning program, termed the Ecogarden Farmer and Biodiversity Surveyor, has been running at the university since 2015. The program is divided into two consecutive phases: a training phase comprising various learning activities and a successive internship phase consisting of the all-round practical tasks involved in managing the garden. A retrospective evaluation of the program using phenomenographic approach and content analysis was adopted to reveal the diversity of students' learning experience, as the indicators of the success of the program. Of 112 participants from 4 cohorts, 32 completed online questionnaires, and semi-structured interviews were successfully conducted with twelve participants, three from each of the four cohorts. The results indicated that the program's outcomes could be categorized into five themes. Most outcomes fit into the theme 'an increase in knowledge and skill level,' followed by 'rise in environmental awareness,''facilitation of personal growth,' and 'enhancement of career development.' Many structural experiences revealed may suggest the success of the program. The longer the participants had participated in the program, the more in-depth and diversified reflection of the senior participants relating to personal development were mentioned. This study provides critical insights into the validity of retrospective program evaluation for assessing the long-term impact of EfS programs by introducing a cross-sectional study of different cohorts as a serial time-point sampling strategy.

Effect of sulfur and sulfur-iron modified biochar on cadmium availability and transfer in the soil-rice system.

Cadmium (Cd) contamination in paddy soils has aroused global concern. Sulfur modified biochar (BC) could combine the benefits of BC and S for Cd remediation. However, no information is available on the impact of sulfur modified biochar on Cd phytoavailability in paddy soils. In this study, a pot experiment was conducted to investigate the effect of sulfur modified biochar (S-BC) and sulfur and iron (Fe) modified biochar (S-Fe BC) on Cd mobility and Cd transfer in the soil-rice system. The application of S-BC and S-Fe BC effectively reduced pore water Cd in the rhizosphere and non-rhizosphere pore water throughout the rice growth stages. S-BC and S-Fe BC addition increased the total chlorophyll content, as well as the root, shoot and grain biomasses of rice. Furthermore, S-BC and S-Fe BC amendments greatly increase the formation of Fe plaque on rice root surface, thus decreasing Cd accumulation in different rice tissues. In particular, S-Fe BC supplementation significantly reduced the Cd concentration in rice grains to 0.018 mg kg-1 in Cd-contaminated soil, which was lower than the China National standard for food contamination limit (0.2 mg kg-1 Cd). Sequential extraction results showed that S-BC and S-Fe BC can promote the transfer of exchangeable Cd to Fe-Mn oxide, organic and residual bound forms which reduce Cd in paddy soils. Thus, the amendment of S-Fe BC to Cd-contaminated paddy soil is an effective strategy to decrease Cd accumulation in rice grains and thereby protect public health.

Effect of sulfur-iron modified biochar on the available cadmium and bacterial community structure in contaminated soils.

Cadmium contamination in paddy soils has aroused increasing concern around the world, and biochar has many positive properties, such as large specific surface areas, micro porous structure for the heavy metal immobilization in soils. However there are few studies on sulfur-iron modified biochar as well as its microbiology effects. The purpose of this study was to evaluate the Cd immobilization effects of sulfur or sulfur-iron modified biochar and its related microbial community changes in Cd-contaminated soils. SEM-EDX analysis confirmed that sulfur and iron were loaded on the raw biochar successfully. Sulfur-modified biochar (S-BC) and sulfur-iron modified biochar (SF-BC) addition increased pH value and the content of soil organic matter, and also decreased DTPA-extractable Cd. There was a negative significant correlation between organic matter content and the available Cd (P < 0.05). During a 45-d incubation period, the fractions of Cd are mainly with the exchangeable (25.16-35.79%) and carbonate (22.01-25.10%) fractions. Compared with the control, the concentrations of exchangeable Cd in soil were significantly (P < 0.05) decreased by 12.54%, 29.71%, 18.53% under the treatments of BC, S-BC, SF-BC respectively. The S-BC and SF-BC treatments significantly (P < 0.05) increased Chao1, observed, Shannon and Simpson diversity indices compared with the control and biochar treatments. Meanwhile, the relative abundance of Proteobacteria, Bacteroidetes, and Actinobacteria increased, whereas the abundance of Acidobacteria and Germmatimonadetes decreased. Capsule: Sulfur-modified and sulfur-iron modified biochar applications decreased the available Cd and changed the microbial community.

Remediation of arsenic-contaminated paddy soil by iron-modified biochar.

Arsenic contamination in paddy soils has aroused global concern due to its threats to food security and human health. Biochar modified with different iron materials was prepared for arsenic (As) immobilization in contaminated soils. Soil incubation experiments were carried to investigate the effects of biochar modified with Fe-oxyhydroxy sulfate (Biochar-FeOS), FeCl3 (Biochar-FeCl3), and zero-valent iron (Biochar-Fe) on the pH, NaHCO3-extractable As concentrations, and the As fractions in soils. The scanning electron microscope and X-ray diffraction analysis demonstrated that iron was successfully loaded onto the surface or embedded into the pores of the biochar. Addition of Biochar-FeOS, Biochar-FeCl3, and Biochar-Fe had no significant effects on the soil pH but significantly decreased the contents of NaHCO3-extractable As in soils by 13.95-30.35%, 10.97-28.39%, and 17.98-35.18%, respectively. Biochar-FeOS, Biochar-FeCl3, and Biochar-Fe treatments decreased the concentrations of non-specifically sorbed and specifically sorbed As fractions in soils, and increased the amorphous and poorly crystalline, hydrated Fe, Al oxide-bound, and residual As fractions. Compared with the other iron-modified biochars, Biochar-FeOS showed the most effective immobilization and has the potential for the remediation of As-contaminated paddy soils.