Use of adapted Aspergillus niger in the bioleaching of spent refinery processing catalyst

Spent refinery processing catalyst is listed as a hazardous waste; the toxicity characteristic leaching procedure (TCLP) extracts of the catalyst are found to contain heavy metals at concentrations exceeding the regulated levels. In the present investigation, Aspergillus niger was adapted to single metal ions Ni, Mo or Al (at 100-2,000 mg/L in steps of 100mg/L) and then to a mixture of Ni, Mo and Al (at a mass ratio of 1:2:6, as approximately present in the spent catalyst). Adaptation experiments with single metals showed that the fungus could tolerate up to 1,000 mg/L Ni, 1,200 mg/L Mo and 2,000 mg/L Al. In the presence of a mixture of these metals, the fungus was able to tolerate up to 100mg/L Ni, 200mg/L Mo and 600 mg/L Al. One-step bioleaching experiments with 1 wt% spent catalyst (of particle size <37 microm) were carried out using un-adapted and various adapted fungal strains. In contrast to the adapted strains, the un-adapted strain showed no growth in the presence of the catalyst. Ni:Mo:Al-adapted strain was the most efficient in the leaching of metals from the catalyst (at 78.5% Ni, 82.3% Mo and 65.2% Al) over 30 days due to its tolerance to the toxic elements at 1 wt%. More importantly, the Ni:Mo:Al-adapted strain was capable of bioleaching up to 3 wt% spent catalyst. The TCLP extracts of the spent catalyst after bioleaching using the Ni:Mo:Al-adapted strain showed the concentrations of Ni and Mo were well within the regulated levels.     

Biosorption behavior of heavy metals in bioleaching process of MSWI fly ash by Aspergillus niger

In this study, it was considered that the biosorption of heavy metals by biomass might occur during the bioleaching of fly ash. This work is focused on the biosorption behavior of Al, Fe, Pb and Zn by Aspergillus niger during the bioleaching process. The fungal biomass was contacted with heavy metals solution which extracted from fly ash by using gluconic acid as leaching agent. The equilibrium time for biosorption was about 120 min. The biosorption experiment data at initial pH 6.5 was used to fit the biosorption kinetics and isotherm models. The results indicated that the biosorption of Al, Fe and Zn by A. niger biomass were well described by the pseudo-first order kinetic model. The pseudo-second order kinetic model was more suitable for that of Pb. The Langmuir isotherm model could well describe the biosorption of Fe, Pb and Zn while the Freundlich model could well describe the biosorption of Al. Furthermore, the biosorption of metal ions decreased evidently in the presence of fly ash as compared to that in the absence of fly ash. This research showed that although the biomass sorption occurred during the bioleaching process, it did not inhibit the removal of Al, Fe, Pb and Zn evidently from fly ash.     

Bioleaching of spent fluid catalytic cracking catalyst using Aspergillus niger

The use of the fungus Aspergillus niger for the bioleaching of heavy metals from spent catalyst was investigated, with fluid catalytic cracking (FCC) catalyst as a model. Bioleaching was examined in batch cultures with the spent catalysts at various pulp densities (1-12%). Chemical leaching was also performed using mineral acids (sulphuric and nitric acids) and organic acids (citric, oxalic and gluconic acids), as well as a mixture of organic acids at the same concentrations as that biogenically produced. It was shown that bioleaching realised higher metal extraction than chemical leaching, with A. niger mobilizing Ni (9%), Fe (23%), Al (30%), V (36%) and Sb (64%) at 1% pulp density. Extraction efficiency generally decreased with increased pulp density. Compared with abiotic controls, bioleaching gave rise to higher metal extractions than leaching using fresh medium and cell-free spent medium. pH decreased during bioleaching, but remained relatively constant in both leaching using fresh medium and cell-free spent medium, thus indicating that the fungus played a role in effecting metal extraction from the spent catalyst.     

Column bioleaching of low-grade mining ore containing high level of smithsonite,talc, sphaerocobaltite and azurite

Present work describes the bioleaching potential of metals from low-grade mining ore containing smithsonite, sphaerocobaltite, azurite and talc as main gangue minerals with adapted consortium of Sulfobacillus thermosulfidooxidans strain-RDB and Thermoplasma acidophilum. Bioleaching potential improved markedly by added energy source, acid preleaching and adaptation of microbial consortium with mixed metal ions. During whole leaching period including acid preleaching stage of 960 h and bioleaching stage of 212 days about 76% Co, 70% Zn, 84% Cu, 72% Ni and 63% Fe leached out.     

Bioleaching of spent refinery processing catalyst using Aspergillus niger with high-yield oxalic acid

A spent refinery processing catalyst was physically and chemically characterized, and subjected to one-step and two-step bioleaching processes using Aspergillus niger. During bioleaching of the spent catalysts of various particle sizes ("as received", 100-150 microm, <37 microm, and x =2.97 (average) microm) and pulp densities, the biomass dry weight and pH were determined. The corresponding leach liquor was analysed for excreted organic acids along with heavy metal values extracted from the catalyst. Chemical characterization of the spent catalyst confirmed the presence of heavy metal including Al (33.3%), Ni (6.09%) and Mo (13.72%). In general, the presence of the spent catalyst caused a decrease in the biomass yield and an increase in oxalic acid secretion by A. niger. The increase in oxalic acid secretion with a decrease in the catalyst particle size (up to <37 microm) led to corresponding increase in the extraction of metal values. The highest extraction of metal values from the spent catalyst (at 1% w/v pulp density and particle size <37 microm) were found to be 54.5% Al, 58.2% Ni and 82.3% Mo in 60 days of bioleaching. Oxalic acid secretion by A. niger in the presence of the spent catalyst was stimulated using 2-[N-Morpholino]ethanesulfonic acid (MES) buffer (pH 6), which resulted in comparable metal extraction (58% Al, 62.8% Ni and 78.9% Mo) in half the time required by the fungus in the absence of the buffer. Spent medium of A. niger grown in the absence and in the presence of MES buffer were found to leach almost similar amounts of Al and Ni, except Mo for which the spent medium of buffered culture was significantly more effective than the non-buffered culture. Overall, this study shows the possible use of bioleaching for the extraction of metal resources from spent catalysts. It also demonstrated the advantages of buffer-stimulated excretion of organic acids by A. niger in bioleaching of the spent catalyst.     

Metal Removal from Contaminated Soils Through Bioleaching with Oxidizing Bacteria and Rhamnolipid Biosurfactants

The use of surfactants as a method for solubilization and removal of heavy metal contamination from soil has been reported before. Biosurfactants produced by some microorganisms are able to modify the surface of various metals and aggregate on interphases favoring the metal separation process from contaminated environments. We evaluated the feasibility of enhancing the removal of metal ions from mineral waste/contaminated soils using alternate cycles of treatment with rhamnolipid biosurfactants and bioleaching with a mixed bacterial culture of Acidithiobacillus thiooxidans and Acidithiobacillus ferrooxidans. Bioleaching alone removed 50% Zn and 19% Fe. When rhamnolipids were used at low concentration (0.4 mg/mL), 11% Fe and 25% Zn were removed, while at 1 mg/mL 19% Fe and 52% Zn removal were achieved. When using a cyclic treatment combining bioleaching and biosurfactants, metal removal reached up to 36% for Fe and 63% to 70% for Zn.     

Remediation of AMD Contaminated Soil by Two Types of Reeds

Acid mine drainage (AMD) adversely impacts many regions in the world. The interactions among citric acid (CA), rhizosphere bacteria and metal uptake in different types of Phragmites australis cultured in spiked AMD contaminated soil were investigated. Compared with non-contaminated reeds cultured under the same conditions, wild reeds harvested from a contaminated site accumulated more metals into tissues. Rhizosphere iron oxidizing bacteria (Fe(II)OB) enhanced the development of Fe plaque but had no significant impact on the formation of Mn and Al plaque on the root surface of either reeds. Plaque may restrain the accumulation of Fe and Mn into tissues of reeds. CA inhibited the growth of Fe(II)OB, reduced the formation of metal plaque and significantly elevated metal accumulations into both underground and aboveground biomass of reeds. The concentrations of Fe, Al and Mn were higher in belowground organs than aboveground tissues. The roots contained 0.28 ± 0.01 mg/g Mn, 3.09 ± 0.51 mg/g Al, 94.47 ± 5.75 mg/g Fe, while the stems accumulated 0.19 ± 0.01 mg/g Mn, 1.34 ± 0.02 mg/g Al, 10.32 ± 0.60 mg/g Fe in wild reeds cultured in soil added with 33,616 ppm CA. Further field investigations may be required to study the effect of CA to enhance phytoremediation of metals from real AMD contaminated sites.     

Metal phytoremediation potential of naturally growing plants on fly ash dumpsite of Patratu thermal power station,Jharkhand, India

Three naturally growing plants Ipomoea carnea, Lantana camara, and Solanum surattense were found in fly ash dumpsite of Patratu thermal power station, Jharkhand, India. They were assessed for their metal uptake potential. The fly ash was slightly alkaline with very less nitrogen and organic carbon but enriched with phosphorus and heavy metals. Lantana camara and Ipomoea carnea showed good translocation from root to shoot for most of the metals except Mn and Pb. The order of metal accumulation in stem of both the plants were Fe(205mg/kg)>Mn(65mg/kg)>Cu(22.35mg/kg)>Pb(6.6mg/kg)>Cr(3.05mg/kg)>Ni(1 mg/kg)>Cd(0.5 mg/kg) and Fe(741 mg/kg)>Mn(154.05 mg/kg)>Cu(20.75 mg/kg)>Pb(6.75 mg/kg)>Ni(4.0 mg/kg)>Cr(3.3mg/kg)>Cd(0.05mg/kg), respectively. But Solanum surattense accumulated most of the metals in roots. The order was in the following order, Mn (382.2mg/kg) >Fe (264.1mg/kg) > Cu (25.35mg/kg) >Pb (5.95 mg/kg) > Ni (1.9 mg/kg) > Cr (1.8mg/kg) > Cd (0.55 mg/kg). The order of Bioconcentration factor (BCF) in root and shoot followed almost the same order as, Mn>Fe>Ni>Pb>Cu>Cr≈ Cd in all the three species. ANOVA showed significant variation in metal accumulation by root and stem between the species. Finally, it can be concluded that Solanum surattense can be used as phytostabilizer and other two species as phytoextractor of metal for fly ash dumpsite reclamation.     

Metal storage in reeds from an acid mine drainage contaminated field

Phragmites australis has been used to treat acid mine drainage (AMD)-contaminated soil. However, the mechanism about metal translocation in reeds was not widely reported. This study investigated metal (Fe, Al, and Mn) storage location in reeds grown in five different sampling sites of an AMD field. As expected, the more metals in soil, the more metals entered the belowground organs of plants. Reeds grown in soils with the highest levels of metals accumulated 0.16 ± 0.04 mg/g Mn, 16.29 ± 4.15 mg/g Fe, and 1.31 ± 0.22 mg/g Al in roots. Most of the iron was sequestered in the roots, while Al was transferred to the shoots. Histological staining found that most of the iron was sequestered in the exodermis, while Al extended the endodermis of roots. Al even entered the stele of roots grown in soil with higher Al levels. The epidermis, cortex, and central cylinder of rhizomes were the main tissues for Fe and Al storage. The more metals in rhizomes, the stronger intensity of the staining was observed around the vascular systems of rhizomes. No structural difference was observed among reeds collected from different sites. Further studies may be needed to enhance the transfer of metals in reeds and increase the phytoremediation efficiency.     

Sulfidogenic fluidized bed treatment of real acid mine drainage water

The treatment of real acid mine drainage water (pH 2.7-4.3) containing sulfate (1.5-3.34 g/L) and various metals was studied in an ethanol-fed sulfate-reducing fluidized bed reactor at 35 °C. The robustness of the process was tested by increasing stepwise sulfate, ethanol and metal loading rates and decreasing feed pH and hydraulic retention time. Highest sulfate reduction rate (4.6 g/L day) was obtained with feed sulfate concentration of 2.5 g/L, COD/sulfate ratio of 0.85 and HRT of 12 h. The corresponding sulfate and COD removal efficiencies were about 90% and 80%, respectively. The alkalinity produced in sulfidogenic ethanol oxidation neutralized the acidic mine water. Highest metal precipitation efficiencies were observed at HRT of 24 h, the percent metal removal being over 99.9% for Al (initial concentration 55 mg/L), Co (9.0 mg/L), Cu (49 mg/L), Fe (435 mg/L), Ni (3.8 mg/L), Pb (7.5 mg/L) and Zn (6.6 mg/L), and 94% for Mn (7.21 mg/L).     

嗜酸异养菌对自养菌Acidithiobacillus ferrooxidans金属离子抗性和生物浸出的影响

【目的】了解嗜酸异养菌在诸如酸性矿坑水(AMD)和生物浸出体系等极端酸性环境中对浸矿微生物产生的影响。【方法】研究由嗜酸异养菌Acidiphilium acidophilum和自养菌Acidithiobacillus ferrooxidans经长期驯化后形成的共培养体系分别在Cd2+、Cu2+、Ni2+和Mg2+胁迫下的稳定性;并将此共培养体系应用于黄铁矿和低品位黄铜矿的生物浸出实验。【结果】在上述4种金属离子分别存在的条件下,异养菌Aph.acidophilum均能促进At.ferrooxidans对亚铁的氧化,提高其对能源利用的效率。共培养体系中的异养菌Aph.acidophilum使At.ferrooxidans对Cu2+的最大耐受浓度(MTC)由2.0 g/L提高到5.0 g/L,而且共培养的细胞数量与2.0 g/L Cu2+条件下生长的At.ferrooxidans纯培养相似。另外,共培养中的At.ferrooxidans对Mg2+的MTC也由12.0 g/L提高到17.0 g/L。生物浸出实验中嗜酸异养菌Aph.acidophilum促进了At.ferrooxidans对黄铁矿样品的浸出,浸出率较其纯培养提高了22.7%;但在含铁量较低的低品位黄铜矿浸出体系中共培养和At.ferrooxidans纯培养的浸出率均低于33%。在加入2.0 g/L Fe2+的低品位黄铜矿浸出体系中,共培养和At.ferrooxidans纯培养的浸出率均得到提高,分别达到52.22%和41.27%。【结论】以上结果表明,Aph.acidophilum与At.ferrooxidans共培养在一定的环境胁迫下仍能保持其稳定性并完成各自的生态功能,并且嗜酸异养菌Aph.acidophilum适合在含铁量较高的浸出体系中与铁氧化细菌共同作用来提高生物浸出的效率。     

液氮保存曲霉菌种的效果

本文报告了以蒸馏水、10%甘油和5%二甲基亚砜(DMSO)为保护剂,用液氮冻结法保存5种8株曲霉的效果,并检测了这些菌分别产生的亚甲基丁二酸、柠檬酸、蛋白酶和糖化酶的生理活性。这些菌在液氮气相(接近-150℃)中保存180天全部保持着生活能力,它们的培养特征和形态特征保留原来的形状。所测定的液氮保存8株菌种的生理活性,除两株糖化酶活力稍有降低外,其它菌株没有明显的变化。     

Bioleaching of copper from chalcopyrite ore by fungi

Microorganisms have been geologically active in mineral formation, mineral diagenesis and sedimentation via direct action of their enzymes or indirectly through chemical action of their metabolic products. This property of microorganisms is being harnessed during the recent years for extraction of metals from their ores, especially from low-grade ores. In the present study bioleaching of copper from its low-grade chalcopyrite ore using 26 isolates of acidophilic fungi is reported. Most of these fungal strains belonged to the genera Aspergillus, Penicillium and Rhizopus. The leaching experiments were conducted in Czepek Dox minimal medium containing 1% (100 mesh) ore with shaking at room temperature for 20 days. Out of these, 4 isolates exhibited significant bioleaching activities. Maximum leaching of copper (78 mg/L) was observed with Aspergillus flavus (DSF-8) and Aspergillus niger (DOF-1). Nutritional and environmental conditions for optimum bioleaching were standardized. Present study indicates the usefulness of acidophilic fungi in bioleaching of copper from its low-grade ores.     

BP神经网络优化微生物浸矿工艺

以低品位黄铜矿溶液为原料,浸矿制备Cu2＋能有效提高低品位黄铜矿的利用价值。基于浸矿过程中存在多因素影响的现象,通过正交试验与神经网络分析方法,对浸矿条件（接种量、矿石品位、Fe2＋添加量及浸矿溶液pH）实行优化。结果表明：在正交试验组中最佳试验结果为浸矿产128．753mg／LCu^2＋;BP神经网络优化后的最佳实验组合为微生物接种量12％、矿石品位0．3％、添加Fe^2＋24g/L及浸矿溶液pH1．7,该条件下验证试验产Cu^2＋ 141．352mg／L,通过正交试验及神经网络优化提高了微生物浸出低品位黄铜矿酸性溶液Cu^2＋的产量。     

Evaluation of metal mobility/immobility in fly ash induced by bacterial strains isolated from the rhizospheric zone of Typha latifolia growing on fly ash dumps

In this investigation, 11 bacterial strains were isolated from the rhizospheric zone of Typha latifolia. All the strains were aerobic, showed positive result with indole production and were able to grow in MacConkey agar. However, four strains were gram positive and others gram negative. These strains were inoculated separately in the fly ash with additional source of carbon to test their ability to increase the bioavailability or immobilization of toxic metals like Cu, Zn, Pb, Cd and Mn. It was observed that most of the bacterial strains either enhanced the mobility of Zn, Fe and Mn or immobilized Cu and Cd. However, there were a few exceptions. For example, in contrast to other bacterial strains, NBRFT6 enhanced immobility of Zn and Fe and NBRFT2 of Mn. On the other hand, in place of immobility induced by most of the bacterial strains, NBRFT8 and NBRFT9 enhanced bioavailability of Cu. However, in case of Cd, all the strains without any exception immobilized this metal. The results also indicated that the mobility/immobility of trace metals from the exchangeable fractions was the specific function of bacterial strains depending upon the several edaphic and environmental factors. Based on the extractability of metals from fly ash, a consortium of high performer bacterial strains will be further used to enhance the phytoextraction of metals from fly ash by metal accumulating plants. On the other hand, bacterial strains responsible for immobilization of metals may be used for arresting their leaching to water bodies.     

Role of Iron-Oxidizing Microorganism in Bioleaching of Heavy Metals From Sewage Sludge Contaminated with Textile Dyes

This article reports the role of indigenous iron-oxidizing microorganisms in bioleaching of heavy metals from anaerobically digested sewage sludge in presence of toxic dyes namely, methylene blue (MB), Remazol black B (RBB) and mixture of both the dyes (DM). To achieve this goal, different concentrations of dyes (0, 500, 10,000, 15,000 and 25,000 mg/L) were added to the sewage sludge (initial pH ～ 7) and subsequently, the solubilization of heavy metals (Cu, Ni, Zn and Cr) was measured at time intervals of 48 h for 16 days. The results showed that an increase in dye concentration adversely affected the population of indigenous iron-oxidizing microorganisms, thereby decreased the bioleaching of metals. The metal solubilization from sludge is more adversely affected in presence of RBB and DM than MB. In majority of cases, the maximum metal solubilization was recorded at dye concentration of 5,000 mg/L. Two factor analyses (ANOVA) suggests the impact of both factors: dye concentration and dye type on metal leaching rate in sludge system.     

Development of a Laboratory-Scale Leaching Plant for Metal Extraction from Fly Ash by Thiobacillus Strains

Semicontinuous biohydrometallurgical processing of fly ash from municipal waste incineration was performed in a laboratory-scale leaching plant (LSLP) by using a mixed culture of Thiobacillus thiooxidans and Thiobacillus ferrooxidans. The LSLP consisted of three serially connected reaction vessels, reservoirs for a fly ash suspension and a bacterial stock culture, and a vacuum filter unit. The LSLP was operated with an ash concentration of 50 g liter⁻¹, and the mean residence time was 6 days (2 days in each reaction vessel). The leaching efficiencies (expressed as percentages of the amounts applied) obtained for the economically most interesting metal, Zn, were up to 81%, and the leaching efficiencies for Al were up to 52%. Highly toxic Cd was completely solubilized (100%), and the leaching efficiencies for Cu, Ni, and Cr were 89, 64, and 12%, respectively. The role of T. ferrooxidans in metal mobilization was examined in a series of shake flask experiments. The release of copper present in the fly ash as chalcocite (Cu₂S) or cuprite (Cu₂O) was dependent on the metabolic activity of T. ferrooxidans, whereas other metals, such as Al, Cd, Cr, Ni, and Zn, were solubilized by biotically formed sulfuric acid. Chemical leaching with 5 N H₂SO₄ resulted in significantly increased solubilization only for Zn. The LSLP developed in this study is a promising first step toward a pilot plant with a high capacity to detoxify fly ash for reuse for construction purposes and economical recovery of valuable metals.     

Assessment of bioaccumulation of heavy metal by Pteris vittata L. growing in the vicinity of fly ash

Pteris vittata L. subsp. vittata, a potential arsenic hyperaccumulator fern, growing naturally in the vicinity offly ash was analyzed for the concentration of nine heavy metals (Fe, Cu, Zn Ni, Al, Cr, Pb, Si, and As) from five different sites around of Kanti Thermal Power Station at Muzaffarpur in Bihar State, India. Metal accumulation in P. vittata was correlated with the level of pollution at five selected sampling sites. The results revealed significantly more accumulation of these metals in the above ground parts of the plant than the parts below ground. Statistical parameters such as the coefficient of variation (CV%) showed a higher for As, Cu, Cr, and a lower one for Fe, Ni, Al. There was high spatial variability in the total metal concentration at different sites. The present study confirmed that P. vittata is a heavy metals accumulator and that it is a highly suitable candidate for phytoremediation of metal contaminated wastelands.     

Spatial distribution of metals in the constructed wetlands

Investigation of the spatial distribution of metals was conducted for two constructed wetlands used as tertiary treatment in Chia Nan University of Pharmacy and Science (CNU) and Metal Processing Industries (MPI) located in Tainan, Taiwan. These two distinguished sites were selected to compare the distribution of metals for constructed wetlands treating different types of wastewater. Along the distance, samples of water, sediment, and macrophytes were analyzed for metals including Al, Cd, Cr, Cu, Fe, Mn, Ni, Pb, and Zn. Additionally, measurements of water quality including temperature, pH, EC, ORP, DO, TSS, BOD, COD, and turbidity were performed. Results show that, at CNU, wastewater contained higher organic consititute (BOD 29.3 +/- 11.7 mg/, COD 46.7 +/- 33.6 mg/L) with low metals content. Wastewater at MPI contained low level of organic consititute (BOD 7.1 +/- 3.3 mg/L, and COD 66.0 +/- 56.5 mg/L) and higher metals content. Metals distribution of both sites showed similar results where metals in the sediments in the inlet zone have greater concentrations than other areas. The constructed wetlands can remove Cd, Cu, Ni, Pb, and Zn. However, there was no removal of Al, Cr, Fe, and Mn. A distance along the constructed wetlands had no effect on metal concentrations in macrophyte and water.