Biosorption of Ni, Cr and Cd by metal tolerant Aspergillus niger and Penicillium sp. using single and multi-metal solution

Fungi including Aspergillus and Penicillium, resistant to Ni2+, Cd2+, and Cr6+ were isolated from soil receiving long-term application of municipal wastewater mix with untreated industrial effluents of Aligarh, India. Metal tolerance in term of minimum inhibitory concentration (MIC) was 125-550 microg/ml for Cd, 300-850 microg/ml for Ni and 300-600 microg/ml for Cr against test fungi. Two isolates, Aspergillus niger and Penicillium sp. were tested for their Cr, Ni and Cd biosorption potential using alkali treated, dried and powdered mycelium. Biosorption experiment was conducted in 100 ml of solution at three initial metal concentrations i.e., 2, 4 and 6 mM with contact time (18 hr) and pretreated fungal biomass (0.1g) at 25 degrees C. Biosorption of all metals was found higher at 4 mM initial metal concentration as compared to biosorption at 2 and 6 mM concentrations. At 4 mM initial metal concentration, chromium biosorption was 18.05 and 19.3 mg/g of Aspergillus and Penicillium biomasses, respectively. Similarly, biosorption of Cd and Ni ions was also maximum at 4 mM initial metal concentration by Aspergillus (19.4 mg/g for Cd and 25.05 mg/g of biomass for Ni) and Penicillium (18.6 mg/g for Cd and 17.9 mg/g of biomass for Ni). In general, biosorption of metal was influenced by initial metal concentration and type of the test fungi. The results indicated that fungi of metal contaminated soil have high level of metal tolerance and biosorption properties.     

Bioremediation of Heavy Metals in Liquid Media Through Fungi Isolated from Contaminated Sources

Wastewater particularly from electroplating, paint, leather, metal and tanning industries contain enormous amount of heavy metals. Microorganisms including fungi have been reported to exclude heavy metals from wastewater through bioaccumulation and biosorption at low cost and in eco-friendly way. An attempt was, therefore, made to isolate fungi from sites contaminated with heavy metals for higher tolerance and removal of heavy metals from wastewater. Seventy-six fungal isolates tolerant to heavy metals like Pb, Cd, Cr and Ni were isolated from sewage, sludge and industrial effluents containing heavy metals. Four fungi (Phanerochaete chrysosporium, Aspegillus awamori, Aspergillus flavus, Trichoderma viride) also were included in this study. The majority of the fungal isolates were able to tolerate up to 400 ppm concentration of Pb, Cd, Cr and Ni. The most heavy metal tolerant fungi were studied for removal of heavy metals from liquid media at 50 ppm concentration. Results indicated removal of substantial amount of heavy metals by some of the fungi. With respect to Pb, Cd, Cr and Ni, maximum uptake of 59.67, 16.25, 0.55, and 0.55 mg/g was observed by fungi Pb3 (Aspergillus terreus), Trichoderma viride, Cr8 (Trichoderma longibrachiatum), and isolate Ni27 (A. niger) respectively. This indicated the potential of these fungi as biosorbent for removal of heavy metals from wastewater and industrial effluents containing higher concentration of heavy metals.     

Biosorption of cobalt by fungi from serpentine soil of Andaman

Fungi belonging to Aspergillus, Mortierella, Paecilomyces, Penicillium, Pythium, Rhizopus and Trichoderma, isolated from serpentine soil of Andaman (India) were screened for cobalt-resistance. Eleven out of total 38 isolated fungi which tolerated > 6.0 mM Co(II) were evaluated for cobalt biosorption using dried mycelial biomass. Maximum Co(II)-loading (1036.5 microM/g, 60 min) was achieved with Mortierella SPS 403 biomass, which removed almost 50% of 4.0 mM cobalt from the aqueous solution. Co(II)-sorption kinetics of Mortierella SPS 403 biomass was fast and appreciable quantities of metal [562.5 microM/g] was adsorbed during first 10 min of incubation. The metal biosorption capacity of the isolate was accelerated with increasing cobalt concentration, while it was reverse with increase of initial biomass. The optimum pH and temperature for Co(II) removal were 7.0 and 30 degrees C, respectively. However, Co(II)-uptake was inhibited in presence of other metals (Pb, Cd, Cu, Ni, Cr and Zn). Freundlich adsorption isotherm appropriately describes Mortierella SPS 403 biomass as an efficient Co(II)-biosorbent.     

Biosorption of nickel and cadmium by metal resistant bacterial isolates from agricultural soil irrigated with industrial wastewater

Agricultural soil irrigated with industrial wastewater (more than two decades) analysed for heavy metals revealed high levels of Fe, Cr, Cu, Zn, Ni and Cd. Out of a total of 40 bacterial isolates obtained from these soils, 17 belonged to the family enterobacteriaceae and 10 were Pseudomonas spp. A maximum MIC of 200 for Cd, 400 for Zn and Cu, 800 for Ni, and 1600 microg/ml for Pb was observed. Biosorption of Ni and Cd studies over a range of metal ion concentrations with Escherichia coli WS11 both in single and bi-metal systems showed that the adsorption of Cd and Ni was dependent on the concentrations and followed the Freundlich adsorption isotherm. The biosorption of Ni increased from 6.96 to 55.31 mg/g of cells, and Cd from 4.96 to 45.37 mg/g of cells at a concentration ranging from 50 to 400 microg/ml after 2h of incubation in a single metal solution. A further increase in incubation time had no significant effect on the biosorption of metals.     

Comparative study of Cd(II) and Cr(VI) biosorption on Staphylococcus xylosus and Pseudomonas sp. in single and binary mixtures

Biosorption of Cd(II) and Cr(VI) ions in single solutions using Staphylococcus xylosus and Pseudomonas sp., and their selectivity in binary mixtures was investigated. Langmuir and Freundlich models were applied to describe metal biosorption and the influence of pH, biomass concentration and contact time was determined. Maximum uptake capacity of cadmium was estimated to 250 and 278 mg g(-1), whereas that of chromium to 143 and 95 mg g(-1) for S. xylosus and Pseudomonas sp., respectively. In binary mixtures with Cd(II) ions as the dominant species, there is a profound selectivity for cadmium biosorption, reaching 96% and 89% for Pseudomonas sp. and S. xylosus, respectively, at 10 mg l(-1) Cd(II) and 5 mg l(-1) Cr(VI). Interesting, when chromium (VI) ions are the dominant species, there is selectivity towards chromium around 92% with S. xylosus only.     

Microbial and plant derived biomass for removal of heavy metals from wastewater

Discharge of heavy metals from metal processing industries is known to have adverse effects on the environment. Conventional treatment technologies for removal of heavy metals from aqueous solution are not economical and generate huge quantity of toxic chemical sludge. Biosorption of heavy metals by metabolically inactive non-living biomass of microbial or plant origin is an innovative and alternative technology for removal of these pollutants from aqueous solution. Due to unique chemical composition biomass sequesters metal ions by forming metal complexes from solution and obviates the necessity to maintain special growth-supporting conditions. Biomass of Aspergillus niger, Penicillium chrysogenum, Rhizopus nigricans, Ascophyllum nodosum, Sargassum natans, Chlorella fusca, Oscillatoria anguistissima, Bacillus firmus and Streptomyces sp. have highest metal adsorption capacities ranging from 5 to 641 mg g(-1) mainly for Pb, Zn, Cd, Cr, Cu and Ni. Biomass generated as a by-product of fermentative processes offers great potential for adopting an economical metal-recovery system. The purpose of this paper is to review the available information on various attributes of utilization of microbial and plant derived biomass and explores the possibility of exploiting them for heavy metal remediation.     

Simultaneous production of pullulan and biosorption of metals by Aureobasidium pullulans strain CH-1 on peat hydrolysate

It was demonstrated that during the growth of Aureobasidium pullulans strain CH-1 on the acid hydrolysate of peat from the Vlasina Lake, the content of metals (Cu, Fe, Zn, Mn, Pb, Cd, Ni and Cr) decreased due to biosorption. The reduction in the metal content was found to be in the range (%): 38.2-62.2, 67.7-97.3, 0.02-62.05, 0.05-23.97, 0.16-4.24, 3.45-51.72, 1.18-35.82, 0.86-44.44, for Cu, Fe, Zn, Mn, Pb, Cd, Ni and Cr, respectively. During this process, the metals were accumulated in the biomass, while pullulan, an extracellular polysaccharide produced by Aureobasidium pullulans strain CH-1, was found not to bind the above-mentioned metals.     

Ternary biosorption studies of Cd(II), Cr(III) and Ni(II) on shelled Moringa oleifera seeds

Competitive biosorption of Cd(II), Cr(III) and Ni(II) on unmodified shelled Moringa oleifera seeds (SMOS) present in ternary mixture were compared with the single metal solution. The extent of adsorption capacity of the ternary metal ions tested on unmodified SMOS was low (10-20%) as compared to single metal ions. SMOS removed the target metal ions in the selectivity order of Cd(II) > Cr(III) > Ni(II). Sorption equilibria, calculated from adsorption data, explained favorable performance of biosorption system. Regeneration of exhausted biomass was also attempted for several cycles with a view to restore the sorbent to its original state.     

Improvement of heavy metal biosorption by mycelial dead biomasses (Rhizopus arrhizus, Mucor miehei and Penicillium chrysogenum): pH control and cationic activation

Abstract: Fungal mycelial by-products from fermentation industries present a considerable affinity for soluble metal ions (e.g. Zn, Cd, Ni, Pb, Cr, Ag) and could be used in biosorption processes for purification of contaminated effluents. In this work the influence of pH on sorption parameters is characterized by measuring the isotherms of five heavy metals (Ni, Zn, Cd, Ag and Pb) with Rhizopus arrhizus biomass under pH-controlled conditions. The maximum sorption capacity for lead was observed at pH 7.0 (200 mg g^-l), while silver uptake was weakly affected. The stability of metal-biosorbent complexes is regularly enhanced by pH neutralization, except for lead. A transition in sorption mechanism was observed above pH 6.0. In addition, comparison of various industrial fungal biomasses ( R. arrhizus, Mucor miehei and Penicillium chrysogenum indicated important variations in zinc-binding and buffering properties (0.24, 0.08 and 0.05 mmol g^−l, respectively). Without control, the equilibrium pH (5.8, 3.9 and 4.0) is shown to be related to the initial calcium content of the biosorbent, pH neutralization during metal adsorption increases zinc sorption in all fungi (0.57, 0.52 and 0.33 mmol g-l) but an improvement was also obtained (0.34, 0.33 and 0.10 mmol g⁻¹) by calcium saturation of the biomass before heavy metal accumulation. Breakthrough curves of fixed bed biosorbent columns demonstrated the capacity of the biosorbent process to purify zinc and lead solutions in continuous-flow systems, and confirmed the necessity for cationic activation of the biosorbent before contact with the heavy-metal solution.     

Metal-binding capacity of arbuscular mycorrhizal mycelium

Experiments with excised mycelium of several Glomus spp. with different histories of exposure to heavy metals were carried out to measure their capacities to bind Cd and Zn. Cd sorption was followed for up to 6 h of incubation to determine its time course relationships. Controls treated with a metabolic inhibitor were included to evaluate whether sorption was due to active uptake or passive adsorption. The effect of ion competition (effects of Ca or Zn on Cd sorption) and general measurements of cation exchange capacity (CEC) of roots and hyphae were also performed. The results showed that AM mycelium has a high metal sorption capacity relative to other microorganisms, and a CEC comparable to other fungi. Metal sorption was rapid (<30 min) and appeared mainly to be due to passive adsorption. Adsorption was highest in a metal-tolerant G. mosseae isolate and intermediate for a fungus isolated from a soil treated with metal-contaminated sludge. The former adsorbed up to 0.5 mg Cd per mg dry biomass, which was three times the binding capacity of non-tolerant fungi, and more than 10 times higher than reported values for, e.g., the commonly used biosorption organism Rhizopus arrhizus. The implications of these results for AM involvement in plant protection against excess heavy metal uptake are discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Metal biosorption capacity of the organic solvent tolerant Pseudomonas fluorescens TEM08

Many kinds of biomass are being tested as a biosorption material for metal removal from the contaminated waters. In the present study the biosorption capacity of an organic solvent tolerant (OST) bacterium was investigated against Cr(VI) and Ni(II). The OST strain of Pseudomonas fluorescens TEM08 was isolated from an oil contaminated soil sample and grown in normal culture conditions (type I) and in the presence of the cyclohexane (type II). Two types of cells were used in the biosorption experiments to compare the organic solvent effect on the biosorption capacity. The biosorption equilibrium was described by Langmuir and Freundlich adsorption isotherms. The value of Q(0) was higher for type I cells (40.8 for Cr(VI); 12.4 for Ni(II)) then the type II (40.7 for Cr(VI); 11.2 for Ni(II)). The adsorption capacity constants (K(F)) of Freundlich model for type I cells and for type II cells were 10.87 and 8.78 for Ni(II) and 13.60 and 10.99 for Cr(VI), respectively.     

Enhancement of the efficiency of Cd phytoextraction using bacterial endophytes isolated from Chromolaena odorata,a Cd hyperaccumulator

Phytoextraction is a technique using a hyperaccumulator to remove heavy metals from soil. The efficiency of heavy metal uptake can be enhanced by the inoculation of endophytes. In this study, we isolated and identified 23 endophytes from Chromolaena odorata, a cadmium (Cd) hyperaccumulator that consisted of 19 bacteria, 2 actinomycetes and 2 fungi. All bacteria and fungi could produce at least 1 plant growth promoting factors. However, only 4 bacterial isolates; Paenibacillus sp. SB12, Bacillus sp. SB31, Bacillus sp. LB51, and Alcaligenes sp. RB54 showed the highest minimum inhibitory concentration (MIC) value (2.9 mM), followed by Exiguobacterium sp.RB51 (1.7 mM). Then, these 5 high-MIC bacteria and 1 low-MIC bacterium, Bacillus sp. LB15 were inoculated onto sunflower grown in soil supplemented with 250 mg/kg of Cd. After 60 days, all inoculated plants accumulated significantly higher Cd concentration than the non-inoculated counterparts, and those inoculated with strain LB51 showed the highest Cd accumulation and growth. Interestingly, strain LB15 with low MIC also enhanced Cd accumulation in plants. The results suggest that these bacteria, particularly strain LB51, could be applied to improve Cd accumulation in plants, and that bacteria with low MIC also have the potential to enhance the efficiency of phytoextraction.     

Ferric reductase, superoxide dismutase and alkaline phosphatase activities in siderophore producing fungi

Enzymes associated with release of iron from internalized ferrated siderophore (ferrisiderophore reductase), with damage to the cell at high iron concentration (superoxide dismutase) and siderophore synthesis (alkaline phosphatase), were examined in 3 test fungi viz., Aspergillus sp. ABp4, Aureobasidium pullulans and Rhizopus sp. Extracellular ferrisiderophore reductase activity was present in all the three fungi, but Aureobasidium pullulans, that showed the highest activity (84.3 microM min(-1)), was the only one to produce intra-cellular ferric reductase (147.9 microM min(-1)). Superoxide dismutase was produced by Aureobasidium pullulans and Rhizopus sp., but not by Aspergillus sp. ABp4, that showed intra-cellular enzyme activity in case of ferric reductase and alkaline phosphatase. Maximum SOD activity was seen in Aureobasidium pullulans both extra-cellularly (93.83 ng ml(-1)) and intra-cellularly (57.14 ng ml(-1)). All the test fungi examined, produced intra-cellular alkaline phosphatase. There was no extracellular alkaline phosphatase. Among the three fungi, Aureobasidium pullulans showed highest alkaline phosphatase activity (129.9 microM min(-1)) and Aspergillus sp. ABp4 the least (76.4 microM min(-1)).     

Hexavalent chromium removal in vitro and from industrial wastes, using chromate-resistant strains of filamentous fungi indigenous to contaminated wastes

Two chromate-resistant filamentous fungi, strains H13 and Ed8, were selected from seven independent fungal isolates indigenous to Cr(VI)-contaminated soil because of their ability to decrease hexavalent chromium levels in the growth medium. Morphophysiological studies identified strain H13 as a Penicillium sp. isolate and Ed8 as an Aspergillus sp. isolate. When incubated in minimal medium with glucose as a carbon source and in the presence of 50 microg/mL Cr(VI), these strains caused complete disappearance of Cr(VI) in the growth medium after about 72 h of incubation. Total chromium concentration in growth medium was constant during culture growth, and no accumulation of chromium in fungal biomass was observed. Quantitative determinations of oxidized and reduced chromium species during the reduction process revealed stoichiometric conversion of Cr(VI) to Cr(III). A decrease in Cr(VI) levels from industrial wastes was also induced by Ed8 or H13 biomass. These results indicate that chromate-resistant filamentous fungi with Cr(VI)-reducing capability could be useful for the removal of Cr(VI) contamination.     

Effect of bacterial inoculation of strains of pseudomonas aeruginosa,alcaligenes feacalis and bacillus subtilis on germination,growth and heavy metal (cd,cr, and ni) uptake of brassica juncea

Bacterial inoculation may influence Brassica juncea growth and heavy metal (Ni, Cr, and Cd) accumulation. Three metal tolerant bacterial isolates (BCr3, BCd33, and BNi11) recovered from mine tailings, identified as Pseudomonas aeruginosa KP717554, Alcaligenes feacalis KP717561, and Bacillus subtilis KP717559 were used. The isolates exhibited multiple plant growth beneficial characteristics including the production of indole-3-acetic acid, hydrogen cyanide, ammonia, insoluble phosphate solubilization together with the potential to protect plants against fungal pathogens. Bacterial inoculation improved seeds germination of B. juncea plant in the presence of 0.1 mM Cr, Cd, and Ni, as compared to the control treatment. Compared with control treatment, soil inoculation with bacterial isolates significantly increased the amount of soluble heavy metals in soil by 51% (Cr), 50% (Cd), and 44% (Ni) respectively. Pot experiment of B. juncea grown in soil spiked with 100 mg kg^?1 of NiCl₂, 100 mg kg^?1 of CdCl₂, and 150 mg kg^?1 of K₂Cr₂O₇, revealed that inoculation with metal tolerant bacteria not only protected plants against the toxic effects of heavy metals, but also increased growth and metal accumulation of plants significantly. These findings suggest that such metal tolerant, plant growth promoting bacteria are valuable tools which could be used to develop bio-inoculants for enhancing the efficiency of phytoextraction.     

Biosorption of cadmium and lead ions by ethanol treated waste baker's yeast biomass

The biosorption of cadmium and lead ions from artificial aqueous solutions using waste baker's yeast biomass was investigated. The yeast cells were treated with caustic, ethanol and heat for increasing their biosorption capacity and the highest metal uptake values (15.63 and 17.49 mg g(-1) for Cd(2+) and Pb(2+), respectively) were obtained by ethanol treated yeast cells. The effect of initial metal concentration and pH on biosorption by ethanol treated yeast was studied. The Langmuir model and Freundlich equation were applied to the experimental data and the Langmuir model was found to be in better correlation with the experimental data. The maximum metal uptake values (qmax, mg g(-1)) were found as 31.75 and 60.24 for Cd(2+) and Pb(2+), respectively. Competitive biosorption experiments were performed with Cd(2+) and Pb(2+) together with Cu(2+) and the competitive biosorption capacities of the yeast biomass for all metal ions were found to be lower than in non-competitive conditions.     

Removal of cadmium(II) ion from aqueous system by dry biomass, immobilized live and heat-inactivated Oscillatoria sp. H1 isolated from freshwater (Mogan Lake)

Oscillatoria sp. H1 (Cyanobacteria, microalgae) isolated from Mogan Lake was used for the removal of cadmium ions from aqueous solutions as its dry biomass, alive and heat-inactivated immobilized form on Ca-alginate. Particularly, the effect of physicochemical parameters like pH, initial concentration and contact time were investigated. The sorption of Cd(II) ions on the sorbent used was examined for the cadmium concentrations within the range of 25-250 mg/L. The biosorption of Cd(II) increased as the initial concentration of Cd(II) ions increased in the medium up to 100 mg/L. Maximum biosorption capacities for plain alginate beads, dry biomass, immobilized live Oscillatoria sp. H1 and immobilized heat-inactivated Oscillatoria sp. H1 were 21.2, 30.1, 32.2 and 27.5 mg/g, respectively. Biosorption equilibrium was established in about 1 h for the biosorption processes. The biosorption was well described by Langmuir and Freundlich adsorption isotherms. Maximum adsorption was observed at pH 6.0. The alginate-algae beads could be regenerated using 50 mL of 0.1 mol/L HCl solution with about 85% recovery.     

Effectivity of host-Rhizobium leguminosarum symbiosis in soils receiving sewage water containing heavy metals

Disposal of sewage water in cultivated soils often containing considerable amount of potentially toxic metals such as Cu, Zn, Ni, Cd, Pb and Cr can be beneficial or harmful to plant growth, rhizobial survival, nodulation and nitrogen fixation. Soil samples from 14 such locations were collected. Symbiotic effectivity of host-Rhizobium leguminosarum symbiosis in these soils was assessed. The total metal contents of Cd, Cu, Zn and Ni in all the 14 samples collected from farmer's fields receiving sewage water ranged between 1.3 and 6.7, 55.8-353.2, 356.0-1028.0 and 90.0-199.7 mg kg(-1) of soil, respectively. In Rohtak 1 soil, levels of Cd, Cu and Zn were highest while Ni was highest in Sonipat 2 soil. The content of available Cd, Cu, Zn and Ni in these soils ranged from 1.0-29.3; 6.2-47.0; 2.4-13.5, respectively, and was 2-9 percent of their total metal contents. All the N2 fixing parameters in pea and Egyptian clover were adversely affected by the presence of heavy metals. Available Cd and Cu contents significantly affected the N contents of pea and Egyptian clover plants, whereas Ni contents were negatively correlated with the plant biomass of pea and Egyptian clover.     

Heavy metal removal in a biosorption column by immobilized M. rouxii biomass

Mucor rouxii biomass was immobilized in a polysulfone matrix. The spherical immobilized biomass beads were packed in a column. The biosorption column was able to remove metal ions such as Pb, Cd, Ni and Zn not only from single-component metal solutions but also from multi-component metal solutions. Column kinetics for metal removal were described by the Thomas model. For single-component metal solutions, the metal removal capacities of the beads for Pb, Cd, Ni and Zn were 4.06, 3.76, 0.36 and 1.36 mg/g, respectively. For a multi-component metal solution containing Cd, Ni and Zn, the capacities were 0.36, 0.31 and 0.40 mg/g for Cd, Ni and Zn, respectively. The adsorbed metal ions were easily desorbed from the beads with 0.05N HNO3 solution. After acid desorption and regeneration with deionized water, the beads could be reused to adsorb metal ions at a comparable capacity.     

Determination of the biosorption heats of heavy metal ions on Zoogloea ramigera and Rhizopus arrhizus

The biosorption of Fe(III), Cr(VI), Pb(II), Cu(II) and Ni(II) ions on Zoogloea ramigera (activated sludge bacterium) and Rhizopus arrhizus (filamentous fungus) has been studied as a function of initial metal ion concentration and temperature. The applicability of the Langmuir model for each metal-microorganism system has been tested at different temperatures. The enthalpy change for the biosorption process has been evaluated by using the Langmuir constant b, related to the energy of adsorption. Thermodynamic parameters indicate the exothermic nature of Cu(II) and Ni(II) biosorption on both microorganisms. Fe(III), Cr(VI) and Pb(II) biosorption is determined to be an endothermic process since increased binding occurs as the temperature is increased in the range 15-45 degrees C.