Isolation and identification of potential fungal species for spent engine lubrication oil remediation in Peninsular Malaysia

Improper disposal and spills of spent engine oil into the environment can result in contamination, which eventually affects humans through the food chain. Mycoremediation is an effective and inexpensive alternative to clean up spent engine oil contamination. In recent work, the potential effectiveness of fungi for degrading spent engine oil was confirmed, with the species identified through molecular identification. Fungi that were able to grow in Bushnell Haas Broth supplied with spent engine oil were identified with the potential to utilize spent engine oil as a carbon source. Six species of fungi namely Penicillium simplicissimum, Aspergillus nidulans, Aspergillus niger, Trichoderma longibrachiatum, Aspergillus ustus, and Aspergillus flavus were successfully identified in this study. Over a course of seven days, P. simplicissimum (21.11 percent) was identified as the most effective fungi in degrading spent engine oil, followed by A. nidulans (17.75 percent), A. niger (15.85 percent), T. longibrachiatum (15.12 percent), A. ustus (15.02 percent), and A. flavus (11.80 percent). As these species of fungi were isolated from the natural environment in Peninsular Malaysia, the potential of using these fungi as mycoremediation of spent engine oil was therefore confirmed.     

Changes in Extractability of Cr and Pb in a Polycontaminated Soil After Bioaugmentation With Microbial Producers of Biosurfactants, Organic Acids and Siderophores

Partly because of the low bioavailability of metals, the soil cleaning-up using phytoremediation is usually time-consuming. In order to enhance the amount of metals at the plant's disposal, the soil bioaugmentation coupled together with phytoextraction is an emerging technology. In this preliminary work, two agricultural soils which mainly differed in their Cr, Hg and Pb contents (LC, low-contaminated soil; HC, high-contaminated soil) were bioaugmented in laboratory conditions by either bacterial (Bacillus subtilis, Pseudomonas aeruginosa, Pseudomonas fluorescens or Ralstonia metallidurans) or fungal inocula (Aspergillus niger or Penicillium simplicissimum) and incubated during three weeks. The LC soil pots bioaugmented with A. niger and P. aeruginosa contained higher concentrations of Cr (0.08 and 0.25 mg.kg⁻¹ dw soil) and Pb (0.25 and 0.3 mg.kg⁻¹ dw soil) in the exchangeable fraction F1 (extraction with MgCl₂) by comparison with the non-bioaugmented soil where neither Cr nor Pb was detected. Conversely, immobilization of Cr and Pb in the soil were observed with the other microorganisms. The soil bioaugmentation not only modified the metal speciation for the most easily extractable fractions but also modified the distribution of metals in the other fractions, to a lesser extent nevertheless. The difference in microbial concentrations between the bioaugmented or not HC soils reached up to 1.8 log units. Thus the microorganisms that we chose for the soil bioaugmentation were competitive towards the indigenous microflora. The PCA analysis showed close positive relationships between the microorganisms which potentially produced siderophores in the soil and the amount of Cr and Pb in the fraction F1.     

Stabilization of heavy metals in MSWI fly ash using silica fume

The objective of this work was to investigate the feasibility and effectiveness of silica fume on stabilizing heavy metals in municipal solid waste incineration (MSWI) fly ash. In addition to compressive strength measurements, hydrated pastes were characterized by X-ray diffraction (XRD), thermal-analyses (DTA/TG), and MAS NMR (²⁷Al and ²⁹Si) techniques. It was found that silica fume additions could effectively reduce the leaching of toxic heavy metals. At the addition of 20% silica fume, leaching concentrations for Cu, Pb and Zn of the hydrated paste cured for 7 days decreased from 0.32 mg/L to 0.05 mg/L, 40.99 mg/L to 4.40 mg/L, and 6.96 mg/L to 0.21 mg/L compared with the MSWI fly ash. After curing for 135 days, Cd and Pb in the leachates were not detected, while Cu and Zn concentrations decreased to 0.02 mg/L and 0.03 mg/L. The speciation of Pb and Cd by the modified version of the European Community Bureau of Reference (BCR) extractions showed that these metals converted into more stable state in hydrated pastes of MSWI fly ash in the presence of silica fume. Although exchangeable and weak-acid soluble fractions of Cu and Zn increased with hydration time, silica fume addition of 10% can satisfy the requirement of detoxification for heavy metals investigated in terms of the identification standard of hazardous waste of China.     

Airborne contaminants at waste treatment plants

Airborne contaminants in Finnish waste treatment plants were studied at two plants using mainly manual sorting and one incineration plant. Air samples were analysed for mesophilic and thermotolerant bacteria, mesophilic fungi, endotoxin, dust and heavy metals (lead, cadmium). The concentrations of mesophilic bacteria in the working air of the waste treatment plants were under 2 × 10⁴ colony forming units (cfu) m⁻³ and the concentrations of mesophilic fungi were under 7 × 10⁴ cfu m⁻³. There was no seasonal variation in the concentrations of bacteria, but the concentrations of fungi were highest in autumn at the waste incineration plant. The concentrations of microbes recorded with two sampling methods, the Andersen and Nuclepore filter methods, differed but were usually of the same order of magnitude. The identified bacteria did not include any potent pathogens. In manual sorting the level of organic dust reached 38 mg m⁻³, whereas the 8-h hygiene limit value for organic dust is 5mg m⁻³ and the 15-min hygiene limit value is 10 mg m⁻³. The endotoxin levels were all below 30 ng m⁻³ and the concentrations of heavy metals were low.     

Centrifugal Bioreactors and Their Application in Remediation

A new approach to the maintenance of large microbial populations for bioremediation purposes has been developed in which a centrifugal bioreactor is used to immobilize microbial populations at extremely high density. The cells are ordered into a three‐dimensional array through which wastewater or groundwater volumes may be flowed, unimpeded by frits or screens. The process methodology is independent of the type, shape, or viability of the individual cells immobilized and, thus, may be adapted to many different bioremediation needs. The utilization of this new process has been explored for three different types of remediation: the removal of heavy metals from wastewater, the aerobic degradation of methyl‐tert‐butyl ether (MTBE), and the anaerobic reduction of nitrate to nitrogen gas. This article discusses the use of centrifugal bioreactors and their application in remediation. © 2001 John Wiley & Sons, Inc.     

Bioprocess technology for coal washery effluent treatment by Lemna minor

Duckweed species are promising macrophytes that can be used in wastewater treatment due to their rapid growth, ease of harvest, low fiber feed potential, and high protein contents. Waste‐water contaminants that are likely to be generated during washing of coal are total suspended solids (TSS), chemical oxygen demand (COD), acidity or alkalinity (pH), and metallic contaminants. Bioprocesses were developed to evaluate the potential of duckweed (Lemna minor) to treat coal washery effluent (CWE) as well as to study the impact on the biochemical changes of the Lemna minor. CWE samples were diluted with distilled water (DW) in different ratios as follows: T1‐CWE:DW(20:80%), T2‐CWE:DW(40:60%), T3‐CWE:DW(60:40%), T4‐CWE:DW (80:20%), T5‐CWE (100%), and Control‐DW(100%). The electrical conductivity of the effluent treated with Lemna minor was 0.035 deciSiemens per meter (dS/m) in the control at Day 10 and substantially higher at treatment ratios of 100, 80, and 60 percent (1.754, 1.842, and 1.631 dS/m). The highest amount of TSS was observed at test ratio T5 (38,834 mg/L), followed by T4 (28,816 mg/L), T3 (26,970 mg/L), T2 (15,320 mg/L), T1 (4,524 mg/L), and control (424 mg/L). Total hardness was higher (820 mg/L) in T4 compared to the control (220 mg/L). Total hardness of the effluent decreased after 30 days of incubation in all the treatment aliquots. The CWE at 20 percent concentration increased the duckweed population and no adverse impacts on its growth were observed. At higher concentration (T5) total mortality of Lemna minor was observed. The chlorophyll production was determined to be inversely proportional to the effluent concentration. Based on this study, it is concluded that Lemna minor can be used for treating CWE after dilution with fresh water.     

Biodegradation of Poly(vinyl alcohol) and Bacterial Cellulose Composites by <Emphasis Type="Italic">Aspergillus niger</Emphasis>

The ability of fungal strains to attack a composite material obtained from poly(vinyl alcohol) (PVA) and bacterial cellulose (BC) is investigated. The fungal strain tested was Aspergillus niger. This fungal strain was able to change not only the polymer surface from smoother to rougher, but also to disrupt the polymer. The degradation results were confirmed by visual observations, scanning electron microscopy (SEM) analyses, X-ray diffraction analyses and FTIR spectra of the film samples. SEM micrographs confirmed the growth of fungi on the composite film surface. The degree of microbial degradation depends on culture medium and on composition of polymeric materials, especially on PVA content. The biodegradation process is accelerated by the presence of glucose in the culture medium as an easily available carbon source.     

Accumulation of Heavy Metals by in vitro cultures of plants

The aim of the project is to study heavy metals accumulation by the selected plants in both laboratory and field conditions. Within the experiments the aspen (Populus tremula × tremuloides), sunflower (Helianthus annuus) and corn (Zea mays) plants were studied. The reasons for this selection were: a fast growth of these plants, an accumulation capacity and an ability to survive in different types of soils. The study was carried out on the aspen plantlets grown in vitro. The plants were exposed to the aqueous solutions having concentrations 0.1 mM, 0.5 mM of Pb²⁺ or Ni²⁺, respectively. The accumulation capacityfor aspen, was about 70% of Pb²⁺ originally present in the solution. The starting concentration of Pb²⁺ (0.5 mM) exhibited no negative impact on the growth. Besides in vitro expositions, a pilot-scale phytoremediation experiment was carried out at the polluted industrial area (Zn – 75000 mg/kg), (Pb – 16000 mg/kg), (Cr – 590 mg/kg), (Cd – 90 mg/kg) and (Cu – 1700 mg/kg).     

Effect of chelating agents in phytoremediation of heavy metals

Chelate‐assisted metal uptake by plants has only recently been discovered in the remediation industry. The simultaneous accumulation of lead, arsenic, copper, and cadmium in plants after application of chelating agents to soil is a promising technology enhancement for phytoremediation. One of the most powerful and commonly used chelating agents is ethylene diamine tetra acetic acid (EDTA), which forms complexes with many of the metal contaminants within the natural environment. This study was conducted to determine the efficiency of an emergent wetland plant species Typha sp. and floating wetland macrophytes such as Pistia sp., Azolla sp., Lemna sp., Salvinia sp., and Eichhornia sp. in phytoremediation of various heavy metals with addition of a chelating agent such as EDTA. EDTA addition to the treatment systems increased the uptake of heavy metals by plants, which was much pronounced with lead and copper. However, the pattern of uptake by plants was similar as that of heavy metals without EDTA amendments. © 2012 Wiley Periodicals, Inc.     

Comparison of leaching characteristics of heavy metals in APC residue from an MSW incinerator using various extraction methods

This study investigates four extraction methods (water extraction, toxicity characteristics leaching procedure (TCLP), modified TCLP with pH control, and sequential chemical extraction (SCE)), each representing different liquid-to-solid (L/S) ratios, pH controls, and types of leachant, and their effects on the leaching concentration of heavy metals in municipal solid waste (MSW) incinerator air pollution control (APC) residue. The results indicated that for extraction with distilled water, the heavy metal leaching concentration (mg/l) decreased with L/S ratio, but the amount of heavy metal released (AHMR), defined as the leached amount of heavy metals to the weight of the tested sample (mg/kg), increased with an increase in L/S ratio, in the range of 2-100. The results also showed that both the leaching concentration and the amount of released metals were strongly pH-dependent in the TCLP and modified TCLP tests. In the case of pHs lower than 6.5, the leaching concentrations of Cd, Pb, Cu, Zn, and Cr decreased with an increase in pH. As pH increased higher than 6.5, Cr and Zn were almost insoluble. Meanwhile, Cd and Cu also showed a similar trend but at pHs of 8.5 and 7.5, respectively. Due to the nature of amphoteric elements, in the case of pHs higher than 7, the Pb leaching concentration increased with increasing pH. In modified TCLP tests with the pH value controlled at the same level as in the SCE test, the heavy metal speciation approached the extractable carbonate bound fraction by the SCE. Both amounts of targeted metals leached from the SCE and modified TCLP tests were much higher than those for the regular TCLP and water extraction tests.     

Uptake of chromium by Aspergillus foetidus

Aspergillus foetidus has the ability to take up chromium during the stationary phase of growth and under growth-nonsupportive conditions. We observed a 97% decrease in hexavalent chromium (initial concentration 5 µg/g) at the end of 92 h of growth, which may be due to its reduction to Cr (III) and/or complexation with organic compounds released due to the metabolic activity of the fungus. Replacement culture studies under growth-nonsupportive conditions revealed that the maximum uptake of Cr (VI) at pH 7.0 is 2 mg/g of dry biomass. At low or high pH values, Cr (VI) uptake is significantly reduced. In addition, the initial rate of total chromium uptake is also enhanced by higher biomass concentrations and the presence of glucose. The results obtained through this investigation indicate the possibility of treating waste effluents containing hexavalent chromium using Aspergillus foetidus.     

Fluxes and Loading of Heavy Metals, Benzo[a]pyrene and Oil Products in Vilnius City

The concentrations of heavy metals Pb, Cd, Cu, Zn and Hg, benzo[a]pyrene and oil products (C₁₅–C₂₈) in bulk (wet and dry) atmospheric deposition in Vilnius city in 2005–2006 were analysed. The highest flux to the ground surface of the city residential area, reaching 1,680 mg m^?2 year^?1, was determined for oil products, which in atmospheric bulk deposition was estimated to be mainly in the form of solid sediments. Among heavy metals, the highest flux was determined for Zn (113.5 mg m^?2 year^?1), while the lowest flux was determined for Hg (0.06 mg m^?2 year^?1). The flux of investigated pollutants ranges from a few times, or for some pollutants, up to one order of magnitude higher at the urban sampling site in comparison to residential or background sites. Some hundred tons of oil products, approximately 52 tons of zinc and a considerably lower amount of mercury, benzo[a]pyrene and cadmium deposit yearly to the ground and water surface of Vilnius city. Metallic constructions related to transport and buildings, automobile exhausts, spills of fuel and lubricants are suggested to be the factors which result in the accumulation of high amounts of heavy metals, oil products and other pollutants on the ground surface of the city.     

Factors affecting polycyclic aromatic hydrocarbon biodegradation by Aspergillus flavus

This study investigated the ability of fungi isolated from highly contaminated soil to biodegrade polycyclic aromatic hydrocarbon (PAH) compounds, as well as the effect of several parameters on the biodegradation ability of these fungi. The isolated fungi were identified using ITS rDNA sequencing and tested using 2,6‐dichlorophinolendophenol to determine their preliminary ability to degrade crude oil. The top‐performing fungi, Aspergillus flavus and Aspergillus fumigatus, were selected to test their ability to biodegrade PAH compounds as single isolates. After 15 days of incubation, A. flavus degraded 82.7% of the total PAH compounds, with the complete degradation of six compounds, whereas A. fumigatus degraded 68.9% of the total PAHs, with four aromatic compounds completely degraded. We also tested whether different temperatures, pH, and nitrogen sources influenced the growth of A. flavus and the degradation rate. The degradation process was optimal at a temperature of 30°C, pH of 5.5, and with nitrogen in the form of yeast extract. Finally, the ability of the fungal candidate, A. flavus, to degrade PAH compounds under these optimum conditions was studied. The results showed that 95.87% of the total PAHs, including 11 aromatic compounds, were completely degraded after 15 days of incubation. This suggests that A. flavus is a potential microorganism for the degradation of PAH compounds in aqueous cultures.     

Enhanced phytoremediation: A study of mycorrhizoremediation of heavy metal–contaminated soil

Arbuscular mycorrhizal fungi (AMF) are microscopic fungi that occur naturally in soil and form a symbiosis with plant roots. By colonizing the roots, the fungus increases plant growth by making soil essential elements like zinc and phosphorus more accessible. AMF can play a role in the phytoremediation of heavy metal–contaminated soil (mycorrhizoremediation). Two research experiments were conducted to evaluate the impact of AMF on the extraction of different heavy metals (arsenic, cadmium, lead, selenium, and zinc) in contaminated soil. A grass mixture composed of Festuca rubra, Festuca eliator, Agropyron repens, and Trifolium repens was used in the experiments, and four different types of AMF were investigated: Glomus intraradices, Glomus mosseae, Glomus etunicatum, and Gigaspora gigantea. The results of the study showed that heavy metal extraction by Glomus intraradices colonized plants was the highest of all four AMF tested and was generally higher than nonmycorrhizal plants, depending on the heavy metal concentration in soil and whether it interacted with other metals in soil. However, metal extraction by AMF colonized grasses reached a plateau after an approximately two‐month period showing no further phytoaccumulation. © 2006 Wiley Periodicals, Inc.     

分步沉淀法处理酸性矿山废水

采用分步沉淀工艺处理酸性矿山废水,考察了工艺条件对废水中有价金属元素回收效果的影响。实验结果表明:Ca(OH)_2为适宜的废水pH调节剂;调节废水pH至4.00左右并投加0.05 mL/L的H_2O_2,可首先去除Fe~(2+)及Fe~(3+),得到富Fe渣(w(Fe)=51.00%);调节废水pH至6.00~6.50,先投加50 mg/L的Na_2S,去除废水中的Cu~(2+),获得富Cu渣(w(Cu)=10.89%),再将Na_2S的投加量增至100 mg/L,去除废水中的Zn与Mn,获得富Zn-Mn渣(w(Cu)=2.37%,w(Mn)=6.79%,w(Pb)=1.61%);进一步调节废水pH至8.40,可去除剩余的Zn、Mn及其他重金属。分步沉淀工艺处理后的废水可达标排放,产生的富Fe渣、富Cu渣及富Zn-Mn渣可直接出售或具有利用价值。分步沉淀工艺可实现有价金属元素的高效回收,大幅度降低废水处理的实际成本,值得工程应用与推广。     

Phytoremediation of zinc and cadmium: A study of arbuscular mycorrhizal hyphae

Arbuscular mycorrhizal fungi (AMF) are microscopic fungi naturally occurring in soil that form a symbiosis with plant roots, producing a highly elaborated hyphal mycelium network in soil.In vitro lab experiments were conducted to determine whether extraradical mycorrhizal hyphae are directly involved in sequestration and uptake of essential zinc and nonessential cadmium by plant roots under toxic concentrations. The research is a continuation of an initial study presented in the Spring 2005 issue of Remediation that focused on the specific role of AMF in the speciation of heavy metals. Thus, this article presents a more expanded view. Results show that zinc and cadmium heavy metals are sequestered and translocated to plant roots via extraradical AMF hyphae. Root/growth media accumulation factors reached 5:1 and 18:1 for zinc and cadmium, respectively. Phytoremediation of heavy metal–contaminated soils can be enhanced by mycorrhizal inoculation. © 2005 Wiley Periodicals, Inc.     

Successful full‐scale enhanced anaerobic dechlorination at a NAPL strength 1,1,1‐TCA source area

Bioremediation of 1,1,1‐trichloroethane (TCA) is more challenging than bioremediation of other chlorinated solvents, such as tetrachloroethene (PCE) and trichloroethene (TCE). TCA transformation often occurs under methanogenic and sulfate‐reducing conditions and is mediated by Dehalobacter. The source area at the project site contains moderately permeable medium sand with a low hydraulic gradient and is approximately 0.5 acre. TCA contamination generally extended to 35 feet, with the highest concentrations at approximately 20 feet. The concentrations then decreased with depth; several wells contained 300 to 600 mg/L of TCA prior to bioremediation. The area of treatment also contained 2 to 30 mg/L of TCE from an upgradient source. Initial site groundwater conditions indicated minimal biotic dechlorination and the presence of up to 20 mg/L of nitrate and 90 mg/L of sulfate. Microcosm testing indicated that TCA dechlorination was inhibited by the site's relatively low pH (5 to 5.5) and high TCA concentration. After the pH was adjusted and TCA concentrations were reduced to less than 35 mg/L (by dilution with site water), dechlorination proceeded rapidly using whey (or slower with sodium lactate) as an electron donor. Throughout the remediation program, increased resistance to TCA inhibition (from 35 to 200 mg/L) was observed as the microbes adapted to the elevated TCA concentrations. The article presents the results of a full‐scale enhanced anaerobic dechlorination recirculation system and the successful efforts to eliminate TCA‐ and pH‐related inhibition. © 2012 Wiley Periodicals, Inc.     

Integrated approach to PCE‐impacted site characterization,site management,and enhanced bioremediation

Tetrachloroethene (PCE)‐ and trichloroethene (TCE)‐impacted sites pose significant challenges even when site characterization activities indicate that biodegradation has occurred naturally. Although site‐specific, regulatory, and economic factors play roles in the remedy‐selection process, the application of molecular biological tools to the bioremediation field has streamlined the assessment of remedial alternatives and allowed for detailed evaluation of the chosen remedial technology. The case study described here was performed at a PCE‐impacted site at which reductive dechlorination of PCE and TCE had led to accumulation of cis‐dichlorethene (cis‐DCE) with concentrations ranging from approximately 10 to 100 mg/L. Bio‐Trap® samplers and quantitative polymerase chain reaction (qPCR) enumeration of Dehalococcoides spp. were used to evaluate three remedial options: monitored natural attenuation, biostimulation with HRC®, and biostimulation with HRC‐S®. Dehalococcoides populations in HRC‐S‐amended Bio‐Traps deployed in impacted wells were on the order of 10³ to 10⁴ cells/bead but were below detection limits in most unamended and HRC‐amended Bio‐Traps. Thus the in situ Bio‐Trap study identified biostimulation with HRC‐S as the recommended approach, which was further evaluated with a pilot study. After the pilot HRC‐S injection, Dehalococcoides populations increased to 10⁶ to 10⁷ cells/bead, and concentrations of cis‐DCE and vinyl chloride decreased with concurrent ethene production. Based on these results, a full‐scale HRC‐S injection was designed and implemented at the site. As with the pilot study, full‐scale HRC‐S injection promoted growth of Dehalococcoides spp. and stimulated reductive dechlorination of the daughter products cis‐DCE and vinyl chloride. © 2008 Wiley Periodicals, Inc.     

Biodegradation of chlorinated organic pesticides endosulfan and chlorpyrifos in soil extract broth using fungi

The present study was designed to screen 20 fungi for their potential to degrade the chlorinated organic pesticides endosulfan and chlorpyrifos. Fungi were first screened for their tolerance to various concentrations of target pesticides using soil extract agar and subsequent degradation studies were performed in soil extract broth containing 25 mg/L of the individual pesticide. Pesticide degradation was evaluated using gas chromatography. Other parameters, such as pH and mycelial weight, were also determined. Based on percent growth inhibition of test fungi and subsequent analysis of EC₅₀ values, the overall results revealed that chlorpyrifos showed significantly more growth inhibition in all tested fungi compared with endosulfan. Trametes hirsuta showed complete degradation of both α‐ and β‐endosulfan isomers and Cladosporium cladosporioides displayed maximum degradation of chlorpyrifos. All test fungi degraded endosulfan more efficiently than chlorpyrifos, except Phanerochaete chrysosporium, Trichoderma harzianum, and Trichoderma virens which showed higher degradation of chlorpyrifos than endosulfan. It was also found that all tested fungi degraded α‐endosulfan more efficiently than β‐endosulfan. Endosulfan sulfate was found to be the major degradation product with all tested fungi. Fungi which showed more endosulfan degradation also produced more endosulfan sulfate. However, less endosulfan sulfate was detected with T. hirsuta and Trametes versicolor, although they degraded endosulfan more efficiently.     

Phosphorus recovery from sewage sludge ash through an electrodialytic process

The electrodialytic separation process (ED) was applied to sewage sludge ash (SSA) aiming at phosphorus (P) recovery. As the SSA may have high heavy metals contents, their removal was also assessed. Two SSA were sampled, one immediately after incineration (SA) and the other from an open deposit (SB). Both samples were ED treated as stirred suspensions in sulphuric acid for 3, 7 and 14 days. After 14 days, phosphorus was mainly mobilized towards the anode end (approx. 60% in the SA and 70% in the SB), whereas heavy metals mainly electromigrated towards the cathode end. The anolyte presented a composition of 98% of P, mainly as orthophosphate, and 2% of heavy metals. The highest heavy metal removal was achieved for Cu (ca. 80%) and the lowest for Pb and Fe (between 4% and 6%). The ED showed to be a viable method for phosphorus recovery from SSA, as it promotes the separation of P from the heavy metals.