The highly effective cadmium-resistant mechanism of Pseudomonas aeruginosa and the function of pyoverdine induced by cadmium.

Pyoverdine (PVD) plays an important role in reducing cadmium (Cd) accumulation in plants. Some Pseudomonas aeruginosa (P. aeruginosa) species can produce PVD under Cd(Π) stress. However, the function of Cd(Π)-induced PVD remains unclear. In this study, we isolated a highly effective Cd(Π)-resistant P. aeruginosa which can secrete PVD under Cd(Π) stress and found that PVD secretion has a dose-dependent relationship with Cd(Π) concentration. PVD can form a PVD-Cd complex with Cd(Π), though the PVD-Cd complex is unable to be adsorbed by the cell or enter the cell, so the complexation of PVD and Cd(Π) impedes Cd(Π) adsorption on the cell surface and alleviates the oxidative stress, lipid peroxidation, and morphological destruction of the cell caused by Cd(Π) and effectively improves the resistance of P. aeruginosa to Cd(Π). In summary, our research results indicate that the Cd(Π) resistance mechanism of P. aeruginosa screened is the complexation of PVD for Cd(Π) and the adsorption of bacteria for Cd(Π); furthermore, PVD plays an important role in improving the Cd(Π)-resistant ability of bacteria. This study provides a deeper understanding of the highly effective Cd(Π) resistance mechanism of P. aeruginosa and the function of Cd(Π)-induced PVD in bacteria.

Formation of Sb2O3 microcrystals by Rhodotorula mucilaginosa.

Antimony (Sb) pollution seriously endangers ecological environment and human health. Microbial induced mineralization can effectively convert metal ions into more stable and less soluble crystalline minerals by extracellular polymeric substance (EPS). In this study, an efficient Sb-resistant Rhodotorula mucilaginosa (R. mucilaginosa) was screened, which can resist 41 mM Sb(III) and directly transform Sb(III) into Sb2O3 microcrystals by EPS. The removal efficiency of R. mucilaginosa for 22 mM Sb(III) reached 70% by converting Sb(III) to Sb2O3. The components of supernatants as well as the effects of supernatants and pH on Sb(III) mineralization verified that inducible and non-inducible extracellular protein/polysaccharide biomacromolecules play important roles in the morphologies and sizes control of Sb2O3 formed by R. mucilaginosa respectively. Sb2O3 microcrystals with different morphologies and sizes can be prepared by the regulation of inducible and non-inducible extracellular biomacromolecules secreted by R. mucilaginosa. This is the first time to identify that R. mucilaginosa can remove Sb(III) by transforming Sb(III) into Sb2O3 microcrystals under the control of EPS. This study contributes to our understanding for Sb(III) biomineralization mechanisms and provides strategies for the remediation of Sb-contaminated environment.

Exploring the role of extracellular polymeric substances in the antimony leaching of tailings by Acidithiobacillus ferrooxidans.

The concentration of Sb bearing tailings in water located in abandoned antimony mines was found to be a big problem, as they contaminate other water resources and entire food chain. Microorganisms were found to be key in tailing leaching and reaction speeding in the presence of extracellular polymeric substances (EPS) produced by bacteria. Herein, we investigated the pattern of the Sb leaching from Sb bearing tailings using Acidithiobacillus ferrooxidans, and analyzed the mechanism of EPS in the leaching process of Sb. To completely and deeply understand the functions of EPS in the bioleaching of antimony tailings, the generation behavior of EPS produced by Acidithiobacillus ferrooxidans (A. ferrooxidans) during bioleaching was characterized by three-dimensional excitation-emission matrix (3D-EEM). Meanwhile, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FT-IR) were used to show the changes of EPS functional groups before and after leaching. Compared with the functional groups in EPS produced by A. ferrooxidans before leaching, the content of hydroxyl and amino groups that reduce high-valent metals to low-valent metals in EPS decreases after leaching, and the carbonyl content increases, corresponding to the ratio of trivalent antimony increased, indicating that EPS could reduce the risk of pentavalent antimony to trivalent one. At the same time, with biological scanning electron microscopy and energy spectrum scanning, the observation of EPS on the mineral surface showed that Sb was adsorbed in the EPS, and the XPS of Sb was fine. Spectral analysis showed that the Sb adsorbed in EPS contained both Sb(III) and Sb(V). Besides, for revealing the influence of EPS in the leaching process of Sb from tailings, this work provided an in-depth understanding of the mechanism of Sb released from tailings under the action of A. ferrooxidans and further provides a basis for the biogeochemical cycle of Sb.

Antimony accumulation in zebrafish (Danio rerio) and its effect on genotoxicity, histopathology, and ultrastructure.

Antimony (Sb) is a toxic metal in aquatic ecosystems. In this study, the accumulation of aqueous Sb in the liver, brain, gills and muscle of zebrafish (Danio rerio) and its effect on genotoxicity, histopathology and ultrastructure alterations were evaluated. The fishes were exposed to different concentrations (0, 8.29, 16.58, 33.16 mg L-1) of aqueous Sb for 18 days. The results showed that the order of Sb accumulation in different tissues was liver > gill > muscle > brain, and the accumulation increased with increasing Sb stress concentration. The mRNA expression levels of Nrf2, Cu/Zn-SOD, Mn-SOD, CAT and GPx genes showed different trends. In addition, significant histopathology and ultrastructure alterations were observed in the liver and gills exposed to Sb. Sb could accumulate in different tissues of zebrafish, inducing the expression of oxidative stress genes and activating antioxidant defense systems. Histological and ultrastructural changes could be used as valid biomarkers for the assessment of aqueous Sb contamination.

Application of biochar immobilized microorganisms for pollutants removal from wastewater: A review.

Microbial immobilization technology (MIT) has been rapidly developed and used to remove pollutants from water/wastewater in recent years, owing to its high stability, rapid reaction rate, and high activity. Microbial immobilization carrier with low cost and high removal efficiency is the key of MIT. Biochar is considered to be an efficient carrier for microbial immobilization because of its high porosity and good adsorption effect, which can provide a habitat for microorganisms. The use of biochar immobilized microorganisms to treat different pollutants in wastewater is a promising treatment method. Compared with the other biological treatment technology, biochar immobilized microorganisms can improve microbial abundance, repeated utilization ratio, microbial metabolic capacity, etc. However, current research on this method is still in its infancy. Little attention has been paid to the interaction mechanisms between biochar and microorganisms, and many studies are only carried out in the laboratory. There are still problems such as difficult recovery after use and secondary pollution caused by residual pollutants after biochar adsorption, which need further clarification. To have comprehensive digestion and an in-depth understanding of biochar immobilized microorganisms technology in wastewater treatment, the wastewater treatment methods based on biochar are firstly summarized in this review. Then the mechanisms of immobilized microorganisms were explored, and the applications of biochar immobilized microorganisms in wastewater were systematically reviewed. Finally, suggestions and perspectives for future research and practical application are put forward.

Role of manganese superoxide dismutase (Mn-SOD) against Cr(III)-induced toxicity in bacteria.

The toxicity of Cr(VI) was widely investigated, but the defense mechanism against Cr(III) in bacteria are seldom reported. Here, we found that Cr(III) inhibited bacterial growth and induced reactive oxygen species (ROS). After exposure to Cr(III), loss of sodA not only led to the excessive generation of ROS, but also enhanced the level of lipid peroxidation and reduced the GSH level, indicating that the deficiency of Mn-SOD decreased the bacterial resistance ability against Cr(III). The adverse effects of oxidative stress caused by Cr(III) could be recovered by the rescue of Mn-SOD in the sodA-deficient strain. Besides the oxidative stress, Cr(III) could cause the bacterial morphology variation, which was distinct between the wild-type and the sodA-deficient strains due to the differential expressions of Z-ring division genes. Moreover, Mn-SOD might prevent Cr(III) from oxidation on the bacterial surface by combining with Cr(III). Taken together, our results indicated that the Mn-SOD played a vital role in regulating the stress resistance, expression of cell division-related genes, bacterial morphology, and chemistry valence state of Cr. Our findings firstly provided a more in-depth understanding of Cr(III) toxicity and bacterial defense mechanism against Cr(III).

Impact of water quality on the microbial diversity in the surface water along the Three Gorge Reservoir (TGR), China.

The Three Gorges Reservoir (TGR), one of the world's largest reservoirs, has crucial roles in flood control, power generation, and navigation. The TGR is contaminated because of the human activities, and how the contaminated water influences the distribution of the microbial community have not been well studied. In this study, we collected 41 freshwater samples from 13 main dwelling districts along the TGR to investigate the water quality, the distribution of the microbial community, and how water quality affects the microbial community structure. The sampling sites cover the whole TGR along the stream, with 670 km distance. Our results show that both water quality and the compositions of bacterial community vary along the TGR. The distribution of bacterial community is closely related to the local water quality. There is the highest concentration of chemical oxygen demand (COD), the highest relative abundance of Firmicutes, and the highest relative abundance of Bacillus in the upstream, compared to the middle and down streams. Redundancy analysis (RDA) showed that PO43- and COD were the main environmental factors influencing on the structure of bacterial community. The relative abundance of nitrification and denitrification functional genes also altered along the streams. These findings provide the basic data for water quality, the distribution of bacterial community, the link of environmental factors, and the bacterial community structure along the TGR, which guides the local environmental protection agency to launch protection strategy for maintaining the ecosystem health of the TGR.

Transcriptomic response and perturbation of toxicity pathways in zebrafish larvae after exposure to graphene quantum dots (GQDs).

Graphene quantum dots (GQDs) are widely used for biomedical applications. Previously, the low-level toxicity of GQDs in vivo and in vitro has been elucidated, but the underlying molecular mechanisms remained largely unknown. Here, we employed the Illumina high-throughput RNA-sequencing to explore the whole-transcriptome profiling of zebrafish larvae after exposure to GQDs. Comparative transcriptome analysis identified 2116 differentially expressed genes between GQDs exposed groups and control. Functional classification demonstrated that a large proportion of genes involved in acute inflammatory responses and detoxifying process were significantly up-regulated by GQDs. The inferred gene regulatory network suggested that activator protein 1 (AP-1) was the early-response transcription factor in the linkage of a cascade of downstream (pro-) inflammatory signals with the apoptosis signals. Moreover, hierarchical signaling threshold determined the high sensitivity of complement system in zebrafish when exposed to the sublethal dose of GQDs. Further, 35 candidate genes from various signaling pathways were further validated by qPCR after exposure to 25, 50, and 100 µg/mL of GQDs. Taken together, our study provided a valuable insight into the molecular mechanisms of potential bleeding risks and detoxifying processes in response to GQDs exposure, thereby establishing a mechanistic basis for the biosafety evaluation of GQDs.

Generation and application of a novel transgenic zebrafish line Tg(cyp1a:mCherry) as an in vivo assay to sensitively monitor PAHs and TCDD in the environment.

The polycyclic aromatic hydrocarbons (PAHs) and 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) are classified as human carcinogens, and can also cause serious health problems. To develop a convenient bio-monitoring tool for the detection of PAHs and TCDD in the environment, we generated a transgenic zebrafish line Tg(cyp1a:mCherry) with cyp1a promoter driving mCherry expression. Here, Tg(cyp1a:mCherry) embryos were treated with different concentrations of TCDD and five US EPA priority PAHs congeners. The results showed that the expressions of mCherry and endogenous cyp1a were consistent with the PAHs exposure concentrations and were largely induced by TCDD and ≥4-ring PAHs. Moreover, the sensitivity of Tg(cyp1a:mCherry) embryos was also evaluated through monitoring of the PAHs contamination in the water and soil samples. The elevated red fluorescent signals and cyp1a expression levels were observed in Tg(cyp1a:mCherry) zebrafish after exposure to water samples and soil organic extracts with higher concentrations of ≥4-ring PAHs. These results further strengthen our findings of concentration- and congener-dependent response of the newly established zebrafish. Taken together, the newly established zebrafish line will prove as a sensitive, efficient and convenient tool for monitoring PAHs and TCDD contamination in the environment.

Perturbation effect of reduced graphene oxide quantum dots (rGOQDs) on aryl hydrocarbon receptor (AhR) pathway in zebrafish.

Graphene quantum dots (GQDs) has been widely used in enormous fields, however, the inherent molecular mechanism of GQDs for potential risks in biological system is still elusive to date. In this study, the outstanding reduced graphene quantum dots (rGOQDs) with the QY as high as 24.62% were successfully synthesized by the improved Hummers method and DMF hydrothermal treatment approach. The rGOQDs were N-doped photoluminescent nanomaterials with functional groups on the surface. The fluorescent bio-imaging was performed by exposing zebrafish in different concentrations of the as-prepared rGOQDs, and the distribution of rGOQDs was successfully observed. Moreover, the developmental toxicity and genotoxicity were evaluated to further investigate the potential hazard of rGOQDs. The result indicated that rGOQDs were responsible for the dose-dependent abnormalities on the development of zebrafish. Since the real-time polymerase chain reaction (RT-PCR) results showed that the expression of cyp1a was the highest expression in the selected genes and significantly up-regulated 8.49 fold in zebrafish, the perturbation of rGOQDs on aryl hydrocarbon receptor (AhR) pathway was investigated by using the Tg(cyp1a:gfp) zebrafish for the first time. The results demonstrated that rGOQDs significantly increased the green fluorescent protein (GFP) expression promoted by cyp1a in a dose-dependent manner, which was also further confirmed by the western blotting. This study offered an opportunity to reveal the potential hazards of in vivo bio-probes, which provided a valuable reference for investigating the graphene-based materials on the disturbance of AhR pathway in biological organisms.

A novel forward osmosis system in landfill leachate treatment for removing polycyclic aromatic hydrocarbons and for direct fertigation.

Landfill leachate (LL) is harmful to aquatic environment because it contains high concentrations of dissolved organic matter, inorganic components, heavy metals, and other xenobiotics. Thus, the remediation of LL is crucial for environmental conservation. Here, a potential application of the forward osmosis (FO) filtration process with ammonium bicarbonate (NH4HCO3) as a draw solution (DS) was investigated to remediate membrane bioreactor-treated LL (M-LL). After the leachate treatment, the toxicity and removal efficiencies of polycyclic aromatic hydrocarbons (PAHs) were evaluated using zebrafish and cultured human cells. The water recovery rate was improved using the current protocol up to 86.6% and 91.6% by both the pressure retarded osmosis (PRO) mode and the forward osmosis (FO) mode. Water flux increased with the increasing DS concentrations, but solution velocities decreased with the operation time. Toxicity tests revealed that the M-LL treated by NH4HCO3 had no toxic effect on zebrafish and human cells. Moreover, green fluorescent protein (GFP) expression in the transgenic zebrafish Tg(cyp1a:gfp) induced by PAHs was very weak compared to the effects induced by untreated M-LL. Since the diluted DS met local safety requirements of liquid fertilizer, it could be directly applied as the liquid fertilizer for fertigation. In conclusion, this novel FO system using NH4HCO3 as the DS provides a cheap and efficient protocol to effectively remove PAHs and other pollutants in LL, and the diluted DS can be directly applied to crops as a liquid fertilizer, indicating that this technique is effective and eco-friendly for the treatment of different types of LL.

Diverse toxicological risks of PAHs in surface water with an impounding level of 175m in the Three Gorges Reservoir Area, China.

The impounding level of 175m for the Three Gorges Reservoir (TGR) is of vital importance for efficient flood control, power generation and convenient navigation in China. However, little is known about the spatial distribution and toxicological risks of major pollutants in the Three Gorges Reservoir Area (TGRA) at that stage. The aim of this study is to probe the ubiquitous polycyclic aromatic hydrocarbons (PAHs) contamination and toxicological impacts in the surface water of the TGRA at the highest water impoundment level of 175m. Our results showed that the Æ©PAHs levels ranged from 83 to 1631ng/L in the upper reaches, 354 to 1159ng/L in the middle reaches, and 23 to 747ng/L in the lower reaches of the TGRA. Source apportionment of PAHs indicated that coal combustion, industrial emissions, heavy traffic, agriculture and shipping activities were the primary sources. Compositional pattern highlighted >85% dominancy of low molecular weight (LMW) PAHs in the reservoir. Risk assessment based on risk quotients (RQs) implied moderate to high ecological risks: the upper reaches>the middle reaches>the lower reaches. However, gene expression profiles portrayed contrary scenario because of the presence of relatively higher footprints of high molecular weight (HMW) PAHs in the middle and the lower reaches, which was confirmed by Cox hazard proportional model. Moreover, the transgenic zebrafish Tg(cyp1a:gfp) induced by PAHs also expressed stronger fluorescent signals in the middle and lower reaches. Taken together, different approaches were employed to firstly reveal the real status of ecological toxicity of PAHs and explore the underlying mechanisms at the highest impounding level of 175m in the TGRA.

In vivo characterization of hair and skin derived carbon quantum dots with high quantum yield as long-term bioprobes in zebrafish.

Carbon quantum dots (CDs) were widely investigated because of their tunable fluorescence properties and low toxicity. However, so far there have been no reports on in vivo functional studies of hair and skin derived CDs. Here, hair derived CDs (HCDs) and skin derived CDs (SCDs) were produced by using human hair and pig skin as precursors. The quantum yields (QYs) of HCDs and SCDs were quite high, compared to citric acid derived CDs (CCDs). HCDs and SCDs possess optimal photostability, hypotoxicity and biocompatibility in zebrafish, indicating that HCDs and SCDs possess the capacity of being used as fluorescence probes for in vivo biological imaging. The long-time observation for fluorescence alternation of CDs in zebrafish and the quenching assay of CDs by ATP, NADH and Fe3+ ions demonstrated that the decaying process of CDs in vivo might be induced by the synergistic effect of the metabolism process. All results indicated that large batches and high QYs of CDs can be acquired by employing natural and nontoxic hair and skin as precursors. To our knowledge, this is the first time to report SCDs, in vivo comparative studies of HCDs, SCDs and CCDs as bioprobes, and explore their mechanism of photostability in zebrafish.

Toxicological characterization of a novel wastewater treatment process using EDTA-Na2Zn as draw solution (DS) for the efficient treatment of MBR-treated landfill leachate.

Landfill leachate has become an important source of environmental pollution in past decades, due to the increase of waste volume. Acute toxic and genotoxic hazards to organisms can be caused by landfill leachate. Thus, how to efficiently recover water from landfill leachate and effectively eliminate combined toxicity of landfill leachate are the most pressing issues in waste management. In this study, EDTA-Na2Zn as draw solution (DS) was used to remove the toxicity of membrane bioreactor-treated landfill leachate (MBR-treated landfill leachate) in forward osmosis (FO) process, and nanofiltration (NF) was designed for recovering the diluted DS. Zebrafish and human cells were used for toxicity assay after the novel wastewater treatment process using EDTA-Na2Zn as DS. Results showed that the water recovery rate of MBR-treated landfill leachate (M-LL) in FO membrane system could achieve 66.5% and 71.2% in the PRO and FO mode respectively, and the diluted DS could be efficiently recovered by NF. Toxicity tests performed by using zebrafish and human cells showed that M-LL treated by EDTA-Na2Zn had no toxicity effect on zebrafish larvae and human cells, but it had very slight effect on zebrafish embryos. In conclusion, all results indicated that EDTA-Na2Zn as DS can effectively eliminate toxicity of landfill leachate and this method is economical and eco-friendly for treatment of different types of landfill leachate.