Exploration of the Vermiculite-Induced Bacterial Community and Co-Network Successions during Sludge-Waste Mushroom Co-Composting.

Vermiculite is a clay mineral with unique physical properties that plays a significant role in plant cultivation, soil remediation, and solid waste management. In this research, we first explored how vermiculite-to-microbe interactions evolved during sludge-waste mushroom residue co-composting. Vermiculite's addition had a substantial impact on the microbial α and ß diversities, significantly changed the microbial community pattern, and strengthened the composting nutrient circulation through the formation of more specialist and generalist species. The microbial community characteristics exhibited common co-networks for resisting composting environment stresses. Vermiculite contributed to enhancing the keystone taxa Proteobacteria and Actinobacteriota and caused the ecological function network to diversify in the warming and maturation phases, with more complexity and tightness in the thermophilic phase (with super-generalist species existing). The enhanced microbial interactions induced by vermiculite possessed a greater capacity to facilitate the metabolisms of carbohydrates and amino acids and cellulolysis, thereby promoting composting humification, and nitrogen retention in the final compost and composting maturity. These findings are helpful for us to understand the biological process mechanisms of the effect of vermiculite additives on composting and contribute to the establishment of a theoretical framework for enhancing the microbial interactions in composting systems by adding vermiculite in practical applications.

Extracellular proteins enhance Cupriavidus pauculus nickel tolerance and cell aggregate formation.

Heavy metal-resistant bacteria secrete extracellular proteins (e-PNs). However, the role of e-PNs in heavy metal resistance remains elusive. Here Fourier Transform Infrared Spectroscopy implied that N-H, C = O and NH2-R played a crucial role in the adsorption and resistance of Ni2+ in the model organism Cuprividus pauculus 1490 (C. pauculus). Proteinase K treatment reduced Ni2+ resistance of C. pauculus underlining the essential role of e-PNs. Further three-dimension excitation-emission matrix fluorescence spectroscopy analysis demonstrated that tryptophan proteins as part of the e-PNs increased significantly with Ni2+ treatment. Proteomic and quantitative real-time polymerase chain reaction data indicated that major changes were induced in the metabolism of C. pauculus in response to Ni2+. Among those lipopolysaccharide biosynthesis, general secretion pathways, Ni2+-affiliated transporters and multidrug efflux play an essential role in Ni2+ resistance. Altogether the results provide a conceptual model for comprehending how e-PNs contribute to bacterial resistance and adsorption of Ni2+.

Microbial extracellular polymeric substances alleviate cadmium toxicity in rice (Oryza sativa L.) by regulating cadmium uptake, subcellular distribution and triggering the expression of stress-related genes.

Cadmium (Cd) accumulation in crops causes potential risks to human health. Microbial extracellular polymeric substances (EPS) are a complex mixture of biopolymers that can bind various heavy metals. The present work examined the alleviating effects of EPS on Cd toxicity in rice and its detoxification mechanism. The 100 µM Cd stress hampered the overall plant growth and development, damaged the ultrastructures of both leaf and root cells, and caused severe lipid peroxidation in rice plants. However, applying EPS at a concentration of 100 mg/L during Cd stress resulted in increased biomass, reduced Cd accumulation and transport, and minimized the oxidative damage. EPS application also enhanced Cd retention in the shoot cell walls and root vacuoles, and actively altered the expression of genes involved in cell wall formation, antioxidant defense systems, transcription factors, and hormone metabolism. These findings provide new insights into EPS-mediated mitigation of Cd stress in plants and help us to develop strategies to improve crop yield in Cd-contaminated soils in the future.

Heterotrophic anodic denitrification coupled with cathodic metals recovery from on-site smelting wastewater with a bioelectrochemical system inoculated with mixed Castellaniella species.

Although Castellaniella species are crucial for denitrification, there is no report on their capacity to carry out denitrification and anode respiration simultaneously in a bioelectrochemical system (BES). Herein, the ability of a mixed inoculum of electricigenic Castellaniella species to perform simultaneous denitrification and anode respiration coupled with cathodic metals recovery was investigated in a BES. Results showed that 500 mg/L NO3--N significantly decreased power generation, whereas 100 and 250 mg/L NO3--N had a lesser impact. The single-chamber MFCs (SCMFCs) fed with 100 and 250 mg/L NO3--N concentrations achieved a removal efficiency higher than 90% in all cycles. In contrast, the removal efficiency in the SCMFCs declined dramatically at 500 mg/L NO3--N, which might be attributable to decreased microbial viability as revealed by SEM and CLSM. EPS protein content and enzymatic activities of the biofilms decreased significantly at this concentration. Cyclic voltammetry results revealed that the 500 mg/L NO3--N concentration decreased the redox activities of anodic biofilms, while electrochemical impedance spectroscopy showed that the internal resistance of the SCMFCs at this concentration increased significantly. In addition, BES inoculated with the Castellaniella species was able to simultaneously perform heterotrophic anodic denitrification and cathodic metals recovery from real wastewater. The BES attained Cu2+, Hg2+, Pb2+, and Zn2+ removal efficiencies of 99.86 ± 0.10%, 99.98 ± 0.014%, 99.98 ± 0.01%, and 99.17 ± 0.30%, respectively, from the real wastewater. Cu2+ was bio-electrochemically reduced to Cu0 and Cu2O, whereas Hg0 and HgO constituted the Hg species recovered via bioelectrochemical reduction and chemical deposition, respectively. Furthermore, Pb2+ and Zn2+ were bio-electrochemically reduced to Pb0 and Zn0, respectively. Over 89% of NO3--N was removed from the BES anolyte during the recovery of the metals. This research reveals promising denitrifying exoelectrogens for enhanced power generation, NO3--N removal, and heavy metals recovery in BES.

Evaluation of fungal community assembly and function during food waste composting with Aneurinibacillus sp. LD3 inoculant.

The objective of this work was to evaluate the fungal community assembly and function during food waste composting with Aneurinibacillus sp. LD3 (LD3) inoculant. Inoculation reduced the content of total organic carbon, moisture content, nitrate nitrogen, and nitrite nitrogen. The LD3 inoculant was able to drive the changes in the assembly of the fungal community. In particular, inoculation with LD3 not only increased the relative abundance of Ascomycota and Trichocomaceae_unclassified for lignocellulose degradation at the mesophilic and cooling stages but also reduced the relative abundances of the opportunistic human pathogen Candida. Saprotroph was the predominant fungal trophic mode in composting, and inoculation with LD3 has a better inactivation effect on animal and plant pathogenic fungi during composting. Furthermore, the variation of the fungal community after inoculation with LD3 was the largest explained by temperature (30.64%). These results implied that LD3 significantly regulated fungal composition and function of food waste composting.

Isolation and characterization of a novel thermotolerant alkali lignin-degrading bacterium Aneurinibacillus sp. LD3 and its application in food waste composting.

The aim of this study was to isolate thermotolerant alkali lignin-degrading bacteria and to investigate their degradation characteristics and application in food waste composting. Two thermotolerant alkali lignin-degrading bacteria isolates were identified as Bacillus sp. LD2 (LD2) and a novel species Aneurinibacillus sp. LD3 (LD3). Compared with strain LD2, LD3 had a higher alkali lignin degradation rate (61.28%) and ligninolytic enzyme activities, and the maximum lignin peroxidase, laccase, and manganese peroxidase activities were 3117.25, 1484.5, and 1770.75 U L-1, respectively. GC-MS analysis revealed that low-molecular-weight compounds such as 4'-hydroxy-3'-methoxy acetophenone, vanillic acid, 1-(4-hydroxy-3,5-dimethoxyphenyl), benzoic acid, and octadecanoic acid were formed in the degradation of alkali lignin by LD3, indicating the cleavage of ß-aryl ether, Cα-Cß bonds, and aromatic rings in lignin. Composting results showed that inoculating LD3 improved the degradation of organic matter by 20.11% and reduced the carbon-to-nitrogen (C/N) ratio (15.66). Additionally, a higher decrease in the content of lignocellulose was observed in the LD treatment. FTIR and 3D-EEM spectra analysis indicated that inoculating LD3 promoted the decomposition of easily available organic substances and lignocellulose and the formation of aromatic structures and humic acid-like substances. In brief, the thermotolerant lignin-degrading bacterium Aneurinibacillus sp. LD3 is effective in degrading lignin and improving the quality of composting.

Exploring the dynamic of microbial community and metabolic function in food waste composting amended with traditional Chinese medicine residues.

The aim of this study was to explore the dynamic of microbial community and metabolic function in food waste composting amended with traditional Chinese medicine residues (TCMRs). Results suggested that TCMRs addition at up to 10% leads to a higher peak temperature (60.5 °C), germination index (GI) value (119.26%), and a greater reduction in total organic carbon (TOC) content (8.08%). 10% TCMRs significantly induced the fluctuation of bacterial community composition, as well as the fungal community in the thermophilic phase. The addition of 10% TCMRs enhanced the abundance of bacterial genera such as Acetobacter, Bacillus, and Brevundimonas, as well as fungal genera such as Chaetomium, Thermascus, and Coprinopsis, which accelerated lignocellulose degradation and humification degree. Conversely, the growth of Lactobacillus and Pseudomonas was inhibited by 10% TCMRs to weaken the acidic environment and reduce nitrogen loss. Metabolic function analysis revealed that 10% TCMRs promoted the metabolism of carbohydrate and amino acid, especially citrate cycle, glycolysis/gluconeogenesis, and cysteine and methionine metabolism. Redundancy analysis showed that the carbon to nitrogen (C/N) ratio was the most significant environmental factor influencing the dynamic of bacterial and fungal communities.

Effect of nickel (II) on the performance of anodic electroactive biofilms in bioelectrochemical systems.

The impact of nickel (Ni2+) on the performance of anodic electroactive biofilms (EABs) in the bioelectrochemical system (BES) was investigated in this study. Although it has been reported that Ni2+ influences microorganisms in a number of ways, it is unknown how its presence in the anode of a BES affects extracellular electron transfer (EET) of EABs, microbial viability, and the bacterial community. Results revealed that the addition of Ni2+ decreased power output from 673.24 ± 12.40 mW/m2 at 0 mg/L to 179.26 ± 9.05 mW/m2 at 80 mg/L. The metal and chemical oxygen demand removal efficiencies of the microbial fuel cells (MFCs) declined as Ni2+ concentration increased, which could be attributed to decreased microbial viability as revealed by SEM and CLSM. FTIR analysis revealed the involvement of various microbial biofilm functional groups, including hydroxyl, amides, methyl, amine, and carboxyl, in the uptake of Ni2+. The presence of Ni2+ on the anodic biofilms was confirmed by SEM-EDS and XPS analyses. CV demonstrated that the electron transfer performance of the anodic biofilms was negatively correlated with the various Ni2+ concentrations. EIS showed that the internal resistance of the MFCs increased with increasing Ni2+ concentration, resulting in a decrease in power output. High-throughput sequencing results revealed a decrease in Geobacter and an increase in Desulfovibrio in response to Ni2+ concentrations of 10, 20, 40, and 80 mg/L. Furthermore, the various Ni2+ concentrations decreased the expression of EET-related genes. The Ni2+-fed MFCs had a higher abundance of the nikR gene than the control group, which was important for Ni2+ resistance. This work advances our understanding of Ni2+ inhibition on EABs, as well as the concurrent removal of organic matter and Ni2+ from wastewater.

Insight into the microbial mechanisms for the improvement of composting efficiency driven by Aneurinibacillus sp. LD3.

This work explored the microbial mechanisms for the improvement of composting efficiency driven by thermotolerant lignin-degrading bacterium Aneurinibacillus sp. LD3 (LD3). Results showed that LD3 inoculant prolonged the thermophilic period by 4 days, improved the final content of humic acid, total phosphorus (TP), nitrogen, potassium and seed germination index. Inoculating LD3 enhanced the relative abundance of thermotolerant and phosphate-solubilizing microbes including the phyla of Proteobacteria, Bacteroidota, Firmicutes, and Actinobacteriota, and the genus of Bacillus, Thermoactinomyces, and Pseudomonas. Metabolic function analysis showed that sequences involved in carbohydrate and amino acid metabolism were boosted, while sequences associated with human disease were reduced after inoculating LD3. Spearman correlation analysis revealed that Aneurinibacillus has a significant positive correlation with temperature, TP, Bacillus, and Thermoactinomyces. This study provides useful information for understanding the microbial mechanisms of LD3 promoting composting efficiency, and reveals the tremendous potential of LD3 in the resource utilization of organic solid wastes.

Expression, purification, characterization and direct electrochemistry of two HiPIPs from Acidithiobacillus caldus SM-1.

High-potential iron-sulfur proteins (HiPIPs) from extremely acidophilic chemolithotrophic non-photosynthetic Acidithiobacillus commonly play a crucial role in ferrous or sulfurous biooxidation. Acidithiobacillus exhibit important industrial applications for bioleaching valuable metals from sulfide ores. In this study, two HiPIP genes from thermophilic Acidithiobacillus caldus SM-1 were cloned and successfully expressed, and their proteins were purified. The proteins displayed a brownish color with an optical absorbance peak at approximately 385 nm and an electronic paramagnetic resonance (EPR) g value of approximately 2.01, which confirmed that the iron-sulfur cluster was correctly inserted into the active site when the proteins were generated in E. coli. The proteins were more thermostable than HiPIPs from mesophilic Acidithiobacillus. The direct electron transfer (DET) between HiPIPs and electrode was achieved by the 2-mercaptopyrimidine (MP) surface-modified gold electrodes; the redox potentials of the HiPIP1 and HiPIP2 measured by cyclic voltammetry were approximately 304.5 mV and 400.5 mV, respectively. The electron transfer rate constant was estimated to be 0.75 s-1 and 0.66 s-1, respectively. The MP/Au electrode and Au electrode showed consistent differences in heterogeneous electron transfer rates and electron transfer resistances. Bioinformatics and molecular simulations further explained the direct electron transfer between the proteins and surface-modified electrode.

Impact of bamboo sphere amendment on composting performance and microbial community succession in food waste composting.

The purpose of this study was to find an economical and effective amendment for improving composting performance and product quality, as well as to analyze the microbial community succession in the whole phase of composting. Therefore, the effect of reusable amendment bamboo sphere on composting performance and microbial community succession during food waste composting was investigated. The results showed that 6% bamboo sphere treatment had the highest degree of polymerization (3.7) and humification index (0.18). Compared with control, 6% bamboo sphere amendment increased total nitrogen (TN), phosphorus (TP) and potassium (TK) contents by 13.61%, 19% and 17.42%, respectively. Furthermore, bamboo sphere enhanced bacterial-fungal diversity and improved microbial community composition by enhancing the relative abundance of thermo-tolerance and lignocellulolytic bacteria and fungi. The five most abundant genera in bamboo sphere composting comprised Bacillus (0-71.47%), Chloroplast-norank (0-47.17%), Pusillimonas (0-33.24%), Acinetobacter (0-27.98%) and unclassified Sphingobacteriaceae (0-22.62%). Linear discriminant analysis effect size showed that Firmicutes, Thermoascaceae and Actinobacteriota, which have a relationship with the decomposition of soluble organic matter and lignocellulose, were significantly enriched in bamboo sphere treatment. Canonical correspondence analysis illustrated that total organic carbon (TOC), TK, and TP were the most important environmental factors on microbial community succession in the two composting systems. Together these results suggest that bamboo sphere as a reusable amendment can shorten maturity period, improve humification degree, increase the contents of nutrient and contribute to the succession of microbial community during food waste composting. These findings provide a theoretical basis for improving the efficiency of food waste composting.

A novel polysaccharides-based bioflocculant produced by Bacillus subtilis ZHX3 and its application in the treatment of multiple pollutants.

A high bioflocculant-producing bacterial strain was identified and named Bacillus subtilis ZHX3. Single-factor experiments suggested that 10 g/L starch and 5 g/L yeast extract were optimal for strain ZHX3 to produce bioflocculant MBF-ZHX3. The maximum flocculating rate reached 95.5%, and 3.14 g/L product was extracted after 3 days of cultivation. MBF-ZHX3 was mainly composed of polysaccharides (77.2%) and protein (14.8%). The polysaccharides contained 28.9% uronic acid and 3.7% amino sugar. Rhamnose, arabinose, galactose, glucose, mannose, and galacturonic acid in a molar ratio of 0.35:1.83:3.09:12.66:0.46:3.81 were detected. MBF-ZHX3 had a molecular weight of 10,028 Da and contained abundant groups (-OH, CO, >PO, C-O-C) contributing to flocculation. Adsorption and bridging was considered as the main flocculation mechanism. MBF-ZHX3 was more effective in decolorizing dyes, removing heavy metals and flotation reagents compared to polyacrylamide. The results implied that MBF-ZHX3 has the potential to substitute polyacrylamide in wastewater treatment because of its excellent biological and environmental benefits.

Recovery of heavy metals from industrial wastewater using bioelectrochemical system inoculated with novel Castellaniella species.

Water pollution is a global issue that has drastically increased in recent years due to rapid industrial development. Different technologies have been designed for the removal of pollutants from wastewater. However, most of these techniques are expensive, generate new waste, and focus solely on metal removal instead of metal recovery. In this study, novel facultative exoelectrogenic strains designated Castellaniella sp. A5, Castellaniella sp. B3, and Castellaniella sp. A3 were isolated from a microbial fuel cell (MFC). These isolates were utilized as pure and mixed culture inoculums in a bioelectrochemical system (BES) to produce bioelectricity and treat simulated industrial wastewater. A single-chamber MFC inoculated with the mixed culture attained the highest electricity generation (i.e., 320 mW/m2 power density and 3.19 A/m2 current density), chemical oxygen demand removal efficiency (91.15 ± 0.05%), and coulombic efficiency (54.81 ± 4.18%). In addition, the BES containing biofilms of the mixed culture achieved the highest Cu, Cr, and Cd removal efficiencies of 99.89 ± 0.07%, 99.59 ± 0.53%, and 99.91 ± 0.04%, respectively. The Cr6+ and Cu2+ in the simulated industrial wastewater were recovered via microbial electrochemical reduction as Cr3+ and Cu0, respectively. However, Cd2+ precipitated as Cd (OH)2 or CdCO3 on the surface of the cathodes. These results suggest that a mixed culture inoculum of Castellaniella sp. A5, Castellaniella sp. B3, and Castellaniella sp. A3 has great potential as a biocatalyst in BES for heavy metals recovery from industrial wastewater.

Deciphering the Endophytic and Rhizospheric Microbial Communities of a Metallophyte Commelina communis in Different Cu-Polluted Soils.

Metallophytes microbiota play a key role in plant growth and resistance to heavy metal stress. Comparing to the well-studied single or some specific plant growth-promoting (PGP) bacterial strains, our current understanding of the structural and functional variations of microbiome of metallophytes is still limited. Here, we systematically investigated the endophytic and rhizosphere bacterial community profiles of a metallophyte Commelina communis growing in different Cu-polluted soils by high-throughput sequencing technology. The results showed that the rhizosphere communities of C. communis exhibited a much higher level of diversity and richness than the endosphere communities. Meanwhile, shifts in the bacterial community composition were observed between the rhizosphere and endosphere of C. communis, indicating plant compartment was a strong driver for the divergence between rhizosphere and endosphere community. Among the environmental factors, soil Cu content, followed by OM, TP and TN, played major roles in shaping the bacterial community structure of C. communis. At the highly Cu-contaminated site, Pseudomonas and Sphingomonas were the predominant genera in the endophytic and rhizospheric bacterial communities, respectively, which might enhance copper tolerance as PGP bacteria. In summary, our findings will be useful to better understand metallophyte-microbe interactions and select suitable bacterial taxa when facilitating phytoremediation.

A high-efficiency Klebsiella variicola H12-CMC-FeS@biochar for chromium removal from aqueous solution.

In polluted groundwater, surface water, and industrial sites, chromium is found as one of the most common heavy metals, and one of the 20 main pollutants in China, which poses a great threat to the ecological environment and human health. Combining biological and chemical materials to treat groundwater contaminated by heavy metals is a promising restoration technology. In this research, Klebsiella variicola H12 (abbreviated as K. variicola) was found to have Cr(VI) reduction ability. A high-efficiency Klebsiella variicola H12-carboxymethyl cellulose (abbreviated as CMC)-FeS@biochar system was established for Cr(VI) removal from aqueous solution. The Scanning Electron Microscope-Energy Dispersive Spectrometer (SEM-EDS), X-ray photoelectron spectroscopy (XPS) results indicated that CMC-FeS was successfully loaded onto the surface of biochar, and K. variicola H12 grew well in the presence of CMC-FeS@biochar with microbial biomass up to 4.8 × 108 cells mL-1. Cr(VI) removal rate of CMC-FeS@biochar system, K. variicola H12 system and K. variicola H12 + CMC-FeS@biochar system were 61.8%, 82.2% and 96.6% respectively. This study demonstrated K. variicola H12-CMC-FeS@biochar system have potential value for efficient removal of Cr(VI) from Cr(VI)-polluted groundwater.

Complete genome sequencing and comparative genomic analyses of Bacillus sp. S3, a novel hyper Sb(III)-oxidizing bacterium.

BACKGROUND: Antimonite [Sb(III)]-oxidizing bacterium has great potential in the environmental bioremediation of Sb-polluted sites. Bacillus sp. S3 that was previously isolated from antimony-contaminated soil displayed high Sb(III) resistance and Sb(III) oxidation efficiency. However, the genomic information and evolutionary feature of Bacillus sp. S3 are very scarce. RESULTS: Here, we identified a 5,436,472 bp chromosome with 40.30% GC content and a 241,339 bp plasmid with 36.74% GC content in the complete genome of Bacillus sp. S3. Genomic annotation showed that Bacillus sp. S3 contained a key aioB gene potentially encoding As (III)/Sb(III) oxidase, which was not shared with other Bacillus strains. Furthermore, a wide variety of genes associated with Sb(III) and other heavy metal (loid) s were also ascertained in Bacillus sp. S3, reflecting its adaptive advantage for growth in the harsh eco-environment. Based on the analysis of phylogenetic relationship and the average nucleotide identities (ANI), Bacillus sp. S3 was proved to a novel species within the Bacillus genus. The majority of mobile genetic elements (MGEs) mainly distributed on chromosomes within the Bacillus genus. Pan-genome analysis showed that the 45 genomes contained 554 core genes and many unique genes were dissected in analyzed genomes. Whole genomic alignment showed that Bacillus genus underwent frequently large-scale evolutionary events. In addition, the origin and evolution analysis of Sb(III)-resistance genes revealed the evolutionary relationships and horizontal gene transfer (HGT) events among the Bacillus genus. The assessment of functionality of heavy metal (loid) s resistance genes emphasized its indispensable role in the harsh eco-environment of Bacillus genus. Real-time quantitative PCR (RT-qPCR) analysis indicated that Sb(III)-related genes were all induced under the Sb(III) stress, while arsC gene was down-regulated. CONCLUSIONS: The results in this study shed light on the molecular mechanisms of Bacillus sp. S3 coping with Sb(III), extended our understanding on the evolutionary relationships between Bacillus sp. S3 and other closely related species, and further enriched the Sb(III) resistance genetic data sources.

Behavior and Mechanism of Cesium Biosorption from Aqueous Solution by Living Synechococcus PCC7002.

Many efforts have focused on the adsorption of metals from contaminated water by microbes. Synechococcus PCC7002, a major marine cyanobacteria, is widely applied to remove metals from the ocean's photic zone. However, its ability to adsorb cesium (Cs) nuclides has received little attention. In this study, the biosorption behavior of Cs(I) from ultrapure distilled water by living Synechococcus PCC7002 was investigated based on kinetic and isotherm studies, and the biosorption mechanism was characterized by Fourier-transform infrared spectroscopy, Raman spectroscopy, scanning electron microscopy, transmission electron microscopy, energy-dispersive X-ray spectrometry, and three-dimensional excitation emission matrix fluorescence spectroscopy. Synechococcus PCC7002 showed extremely high tolerance to Cs ions and its minimal inhibitory concentration was 8.6 g/L. Extracellular polymeric substances (EPS) in Synechococcus PCC7002 played a vital role in this tolerance. The biosorption of Cs by Synechococcus PCC7002 conformed to a Freundlich-type isotherm model and pseudo-second-order kinetics. The binding of Cs(I) was primarily attributed to the extracellular proteins in EPS, with the amino, hydroxyl, and phosphate groups on the cell walls contributing to Cs adsorption. The biosorption of Cs involved two mechanisms: Passive adsorption on the cell surface at low Cs concentrations and active intracellular adsorption at high Cs concentrations. The results demonstrate that the behavior and mechanism of Cs adsorption by Synechococcus PCC7002 differ based on the Cs ions concentration.

Role of extracellular polymeric substance (EPS) in toxicity response of soil bacteria Bacillus sp. S3 to multiple heavy metals.

Heavy metal resistant bacteria are of great interest because of their potential use in bioremediation. Understanding the survival and adaptive strategies of these bacteria under heavy metal stress is important for better utilization of these bacteria in remediation. The objective of this study was to investigate the role of bacterial extracellular polymeric substance (EPS) in detoxifying against different heavy metals in Bacillus sp. S3, a new hyper antimony-oxidizing bacterium previously isolated from contaminated mine soils. The results showed that Bacillus sp. S3 is a multi-metal resistant bacterial strain, especially to Sb(III), Cu(II) and Cr(VI). Toxic Cd(II), Cr(VI) and Cu(II) could stimulate the secretion of EPS in Bacillus sp. S3, significantly enhancing the adsorption and detoxification capacity of heavy metals. Both Fourier transform infrared spectroscopy (FTIR) and three-dimensional excitation-emission matrix (3D-EEM) analysis further confirmed that proteins were the main compounds of EPS for metal binding. In contrast, the EPS production was not induced under Sb(III) stress. Furthermore, the TEM-EDX micrograph showed that Bacillus sp. S3 strain preferentially transported the Sb(III) to the inside of the cell rather than adsorbed it on the extracellular surface, indicating intracellular detoxification rather than extracellular EPS precipitation played an important role in microbial resistance towards Sb(III). Together, our study suggests that the toxicity response of EPS to heavy metals is associated with difference in EPS properties, metal types and corresponding environmental conditions, which is likely to contribute to microbial-mediated remediation.

Extraction and characterization of extracellular polymeric substances from a mixed fungal culture during the adaptation process with waste printed circuit boards.

Extracellular polymeric substances (EPSs) extracted from fungal mycelium by four chemical methods (NaOH, H2SO4, formaldehyde-NaOH, glutaraldehyde-NaOH), three physical methods (heating, ultrasound, vibration), and a control method (centrifugation alone) were investigated. Results indicated formaldehyde-NaOH outperformed other methods with 186.6 ± 8.0 mg/g of polysaccharides and 23.2 ± 4.6 mg/g of protein extracted and ensured little contamination by intracellular substances. Thereafter, this method was applied in extracting EPS from a mixed fungal culture in the adaptation process with 0.5% (w/v) waste printed circuit boards (PCBs). With the four adaptation stages continuing, the culture tended to become more sensitive to respond to the external toxic environment characterized by secreting EPS more easily and quickly. The maximum amount of polysaccharides and protein could be achieved in only 3 days both at the 3rd and 4th adaptation stage. Three-dimensional excitation-emission matrix fluorescence spectrum indicated the peaks obtained for EPS were mainly associated to soluble microbial by-product-like and aromatic protein-like compounds. Transmission electron microscopic observation illustrated that although metal ions penetrated into hypha cells, parts of them could be absorbed by EPS, implying that EPS secretion may be a primary protective strategy adopted by the culture.

The potential role of brassinosteroids (BRs) in alleviating antimony (Sb) stress in Arabidopsis thaliana.

Brassinosteroids (BRs) play a crucial role in improving plant resistance to various environmental stresses. In this study, we aimed to explore the potential role of BRs in protecting plants from antimony (Sb) toxicity. In the in vitro agar-plate culture experiments, the level changes of BR in wide-type plants and BR biosynthesis mutant dwrf4-1 significantly affected the corresponding response of Arabidopsis to Sb stress. Increasing the BR content significantly enhanced Sb-induced root growth inhibition and lowering the BR level appeared to reduce the plant sensitivity to Sb stress. Foliar application of eBL, however, significantly decreased the Sb accumulation and peroxidation of membrane lipids, increased the contents of chlorophyll and proline, and further boosted and strengthened the antioxidant enzymes activities. These experiments demonstrated that BRs played an important role in regulating heavy metal stress responses in plants and exogenous foliar spray of eBL was an important method for alleviating toxicity of Sb.