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.

DNA aptamers specific for Legionella pneumophila: systematic evolution of ligands by exponential enrichment in whole bacterial cells.

Legionella pneumophila is the major causative agent of Legionnaires' disease and Pontiac fever, which pose major public health problems. Rapid detection of L. pneumophila is important for global control of these diseases. Aptamers, short oligonucleotides that bind to targets with high affinity and specificity, have great potential for use in pathogenic bacterium detection, diagnostics, and therapy. Here, we used a whole-cell SELEX (systematic evolution of ligands by exponential enrichment) method to isolate and characterize single-stranded DNA (ssDNA) aptamers against L. pneumophila. A total of 60 ssDNA sequences were identified after 17 rounds of selection. Other bacterial species (Escherichia coli, Bacillus subtilis, Pseudomonas syringae, Staphylococcus aureus, Legionella quateirensis, and Legionella adelaidensis) were used for counterselection to enhance the specificity of ssDNA aptamers against L. pneumophila. Four ssDNA aptamers showed strong affinity and high selectivity for L. pneumophila, with Kd values in the nanomolar range. Bioinformatic analysis of the most specific aptamers revealed predicted conserved secondary structures that might bind to L. pneumophila cell walls. In addition, the binding of these four fluorescently labeled aptamers to the surface of L. pneumophila was observed directly by fluorescence microscopy. These aptamers identified in this study could be used in the future to develop medical diagnostic tools and public environmental detection assays for L. pneumophila.

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.

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.

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.

Insights into the production of extracellular polymeric substances of Cupriavidus pauculus 1490 under the stimulation of heavy metal ions.

Three different methods (a sulfuric acid method, sodium chloride method and vibration method) were used to extract extracellular polymeric substances (EPS) from Cupriavidus pauculus 1490 (C. pauculus 1490) in the present study. The sodium chloride method was able to extract the maximum amount of EPS (86.15 ± 1.50 mg g-1-DW), and could ensure minimum cell lysis by detecting glucose-6-phosphate dehydrogenase activity and using scanning electron microscopy. This method was therefore selected as the optimal extraction method and used in subsequent experiments. On this basis, the tolerance of C. pauculus 1490 and variations in EPS secretion after the addition of different metal ions was investigated. The tolerance levels of C. pauculus 1490 to Cd(ii), Ni(ii), Cu(ii) and Co(ii) were 300 mg L-1, 400 mg L-1, 400 mg L-1 and 400 mg L-1, respectively. Low concentrations of these heavy metal ions could promote bacterial growth, while increased concentrations were found to inhibit it. The results show that metal ions, especially Cd(ii), stimulate the secretion of EPS, with an EPS yield reaching 956.12 ± 10.59 mg g-1-DW at 100 mg L-1. Real-time polymerase chain reaction (PCR) analysis showed that the key EPS synthetic genes, epsB, epsP and Wzz, were up-regulated. Fourier transform infrared spectroscopy analysis suggested that abundant functional groups in EPS play an important role in heavy metal ion complexation. These results will contribute to our understanding of the tolerance mechanism of microorganisms in the presence of different types and concentrations of metal ions.

Application of the kinetic and isotherm models for better understanding of the mechanism of biomineralization process induced by Purpureocillium lilacinum Y3.

Purpureocillium lilacinum can promote the biomineralization of jarosite by secreting extracellular polymeric substances (EPS), but the detailed mechanism is not clear. In this study, the biosynthesis process of jarosite induced by P. lilacinum Y3 and hypha cell surface characterization were investigated. X-ray diffraction (XRD) analysis indicated that P. lilacinum Y3 could induce the formation of jarosite crystal and enhance mineralization kinetics. The kinetic and isotherm models confirmed that the metal ions transferring from the solution to the mycelium surface was controlled by diffusion process and the active interfacial sites on hypha cell surface played a pivotal role in the biomineralization process. Furthermore, transmission electron microscopy (TEM) pictures illustrated that the P. lilacinum Y3 mainly induced the generation of mineral precipitate extracellularly, but not intracellularly. Three-dimensional excitation-emission matrix (3D-EEM) fluorescence spectrum results further revealed the extracellular compounds such as fulvic-acid-like and protein-like substances participated in the mineralization process.

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.

Extracellular polymeric substances (EPS) secreted by Purpureocillium lilacinum strain Y3 promote biosynthesis of jarosite.

In this study, the biosynthesis of jarosite by Purpureocillium lilacinum was investigated. Firstly, we found when the pH value was lower than 2.50 at 30 °C, the concentration of Fe3+ in the solution significantly dropped about 72% after inoculation and a yellow-ocher precipitate was observed on the mycelium surface. X-ray diffraction analysis revealed the precipitate was jarosite. Thereafter, the characterization of the biomineralization process by scanning electron microscopy showed that mineral precipitates started on the cell surface, and then thoroughly covered it. Furthermore the effect of extracellular polymeric substances (EPS) on the biosynthesis of jarosite was investigated. The results suggested Fe3+ only dropped 5.2% in 2 days when EPS were stripped. Finally, through monitoring the changes of mycelium surface groups by Fourier transform infrared spectroscopy, we found the biomineralization process originated from the existence of free P[double bond, length as m-dash]O groups in EPS which acted as crystallization nuclei to promote Fe(OH)3 transformation into jarosite by the formation of P-O-Fe bonds.

Isolation and identification of Penicillium chrysogenum strain Y5 and its copper extraction characterization from waste printed circuit boards.

Biohydrometallurgy is generally considered as a green technology for the recycling of industrial solid waste. In this study, an indigenous fungal strain named Y5 with the ability of high-yielding organic acids was isolated and applied in bioleaching of waste printed circuit boards (PCBs). The strain Y5 was identified as Penicillium chrysogenum by morphological and molecular identification. Meanwhile, we investigated that an optimal set of culturing conditions for the fungal growth and acids secretion was 15 g/L glucose with initial pH 5.0, temperature 25°C and shaking speed 120 rpm in shaken flasks culture. Moreover, three bioleaching processes such as one-step, two-step and spent medium processes were conducted to extract copper from waste PCBs. Spent medium bioleaching showed higher copper extraction percentage and it was 47% under 5%(w/v) pulp density. Transmission electron microscope (TEM) observation combining with energy dispersive analysis of X-rays (EDAX) showed that the leached metal ions did not obviously damage the hypha cells. All above results indicated that P.chrysogenum strain Y5 has the tolerance to metal ions, suggesting its potential in recycling of metals from waste PCBs in industry.

Recycling of metals from pretreated waste printed circuit boards effectively in stirred tank reactor by a moderately thermophilic culture.

To seek a feasible technique for processing waste printed circuit boards (PCBs), pretreatment of PCBs by table separation and further bioleached by moderate thermophiles in a stirred tank reactor were investigated. The shaking table separation, conducted after grinding and sieving of PCBs, produced two fractions: metal-rich parts (RPCBs), which is more suitable for pyrometallurgy process than untreated PCBs, and metal-poor parts (PPCBs) with only 8.83% metals was then bioleached by a mixed culture of moderate thermophiles effectively. After adaptation, the mixed culture could tolerate 80 g/L PPCBs. The bioleaching results showed that metals recovery was 85.23% Zn, 76.59% Cu and 70.16% Al in only 7 days. Trace Pb and Sn were detected in the leachate because of precipitating. The microorganism community structure was analyzed by amplified ribosomal DNA restriction analysis. Two moderately thermophilic bacteria species were identified as Leptospirillum ferriphilum and Acidithiobacillus caldus. Furthermore, uncultured Thermoplasmatales archaeon was also detected in the leaching system. It was also shown that moderate thermophiles revealed best bioleaching ability when compared with mesophiles and the mixture of mesophiles and moderate thermophiles. Finally, we designed a two-stage process model according to the present study to achieve semi-industrial waste PCBs recycling and economic feasibility analysis indicated that the process was profitable.