The efficiency of sodium hydroxide treated rubber (Hevea brasiliensis) leaves powder (NHBL) for removing copper ions from aqueous solutions has been investigated. The effects of physicochemical parameters on biosorption capacities such as stirring speed, pH, biosorbent dose, initial concentrations of copper, and ionic strength were studied. The biosorption capacities of NHBL increased with increase in pH, stirring speed and copper concentration but decreased with increase in biosorbent dose and ionic strength. The isotherm study indicated that NHBL fitted well with Langmuir model compared to Freundlich and Dubinin-Radushkevich models. The maximum biosorption capacity determined from Langmuir isotherm was 14.97 mg/g at 27 degrees C. The kinetic study revealed that pseudosecond order model fitted well the kinetic data, while Boyd kinetic model indicated that film diffusion was the main rate determining step in biosorption process. Based on surface area analysis, NHBL has low surface area and categorized as macroporous. Fourier transform infrared (FT-IR) analyses revealed that hydroxyl, carboxyl, and amino are the main functional groups involved in the binding of copper ions. Complexation was one of the main mechanisms for the removal of copper ions as indicated by FT-IR spectra. Ion exchange was another possible mechanism since the ratio of adsorbed cations (Cu2+ and H+) to the released cations (Na+, Ca2+, and Mg2+) from NHBL was almost unity. Copper ions bound on NHBL were able to be desorbed at > 99% using 0.05 mol/L HCl, 0.01 mol/L HNO3, and 0.01 mol/L EDTA solutions.
Most components of petroleum oily sludge (POS) are toxic, mutagenic and cancer-causing. Often bioremediation using microorganisms is hindered by the toxicity of POS. Under this circumstance, phytoremediation is the main option as it can overcome the toxicity of POS. Cajanus cajan a legume plant, was evaluated as a phyto-remediating agent for petroleum oily sludge-spiked soil. Culture dependent and independent methods were used to determine the rhizosphere microorganisms' composition. Degradation rates were estimated gravimetrically. The population of total heterotrophic bacteria (THRB) was significantly higher in the uncontaminated soil compared to the contaminated rhizosphere soil with C. cajan, but the population of hydrocarbon-utilizing bacteria (HUB) was higher in the contaminated rhizosphere soil. The results show that for 1 to 3% oily sludge concentrations, an increase in microbial counts for all treatments from day 0 to 90 d was observed with the contaminated rhizosphere CR showing the highest significant increase (p
Spent Pleurotus sajor-caju compost mixed with livestock excreta, i.e. cow dung or goat manure, was contaminated with landfill leachate and vermiremediated in 75 days. Results showed an extreme decrease of heavy metals, i.e. Cd, Cr and Pb up to 99.81% removal as effect of vermiconversion process employing epigeic earthworms i.e. Lumbricus rubellus. In addition, there were increments of Cu and Zn from 15.01% to 85.63%, which was expected as non-accumulative in L. rubellus and secreted out as contained in vermicompost. This phenomenon is due to dual effects of heavy metal excretion period and mineralisation. Nonetheless, the increments were 50-fold below the limit set by EU and USA compost limits and the Malaysian Recommended Site Screening Levels for Contaminated Land (SSLs). Moreover, the vermicompost C:N ratio range is 20.65-22.93 and it can be an advantageous tool to revitalise insalubrious soil by acting as soil stabiliser or conditioner.
Co-composting of poultry manure and rubber wood sawdust was performed with the ratio of 2:1 (V/V) for a period of 60 days. An investigation was carried out to study the extracellular enzymatic activities and structural degradation utilizing Fourier transform infrared spectroscopy (FT-IR), thermogravimetry and differential thermal analysis (TG/DTA) and scanning electron microscopy (SEM). The microbial succession was also determined by using denaturing gel gradient electrophoresis (DGGE). The compost was able to reach its highest temperature of 71°C at day 3 and stabilized between 30 and 40°C for 8 weeks. CMCase, FPase and β-glucosidase acted synergistically in order to degrade the cellulosic substrate. The xylanase activities increased gradually during the composting and reached the peak value of 11.637 U/g on day 35, followed by a sharp decline. Both LiP and MnP activities reached their peak values on day 35 with 0.431 and 0.132 U/g respectively. The FT-IR spectra revealed an increase in aromaticity and a decrease in aliphatic compounds such as carbohydrates as decomposition proceeded. TGA/DTG data exhibited significant changes in weight loss in compost samples, indicating degradation of organic matter. SEM micrographs showed higher amounts of parenchyma exposed on the surface of rubber wood sawdust at day 60, showing significant degradation. DGGE and 16S rDNA analyses showed that Burkholderia sp., Pandoraea sp., and Pseudomonas sp. were present throughout the composting process. Ornithinibacillus sp. and Castellaniella ginsengisoli were only found in the initial stage of the composting, while different strains of Burkholderia sp. also occurred in the later stage of composting.
Indian jujuba seed powder (IJSP) has been investigated as a low-cost and an eco-friendly biosorbent, prepared for the removal of Acid Blue 25 (AB25) from aqueous solution. The prepared biomaterial was characterized by using FTIR and scanning electron microscopic studies. The effect of operation variables, such as IJSP dosage, contact time, concentration, pH, and temperature on the removal of AB25 was investigated, using batch biosorption technique. Removal efficiency increased with increase of IJSP dosage but decreased with increase of temperature. The equilibrium data were analyzed by the Langmuir and the Freundlich isotherm models. The data fitted well with the Langmuir model with a maximum biosorption capacity of 54.95 mg g(-1). The pseudo-second-order kinetics was the best for the biosorption of AB25 by IJSP, with good correlation. Thermodynamic parameters such as standard free energy change (ΔG(0)), standard enthalpy changes (ΔH(0)), and standard entropy changes (ΔS(0)) were analyzed. The removal of AB25 from aqueous solution by IJSP was a spontaneous and exothermic adsorption process. The results suggest that IJSP is a potential low-cost and an eco-friendly biosorbent for the AB25 removal from synthetic AB25 wastewater.
The cell-free extract of locally isolated Rhodococcus UKMP-5M strain was used as an alternative to develop greener and cost effective cyanide removal technology. The present study aims to assess the viability of the cell-free extract to detoxify high concentrations of cyanide which is measured through the monitoring of protein concentration and specific cyanide-degrading activity. When cyanide-grown cells were subjected to grinding in liquid nitrogen which is relatively an inexpressive and fast cell disruption method, highest cyanide-degrading activity of 0.63 mM min(-1) mg(-1) protein was obtained in comparison to enzymatic lysis and agitation with fine glass beads. The cell-free extracts managed to degrade 80% of 20 mM KCN within 80 min and the rate of cyanide consumption increased linearly as the concentration of protein was raised. In both cases, the addition of co-factor was not required which proved to be advantageous economically. The successful formation of ammonia and formate as endproducts indicated that the degradation of cyanide by Rhodococcus UKMP-5M proceeded via the activity of cyanidase and the resulting non-toxic products are safe for disposal into the environment. Further verification with SDS-PAGE revealed that the molecular weight of the active enzyme was estimated to be 38 kDa, which is consistent with previously reported cyanidases. Thus, the utilization of cell-free extracts as an alternative to live microbial in cyanide degradation offers numerous advantageous such as the potential to tolerate and degrade higher concentration of cyanide and total reduction in the overall cost of operation since the requirement for nutrient support is irrelevant.
Free laccase and fungal biomass from white-rot fungi were compared in the thermokinetics study of the laccase-catalyzed decolorization of an azo dye, i.e., Trypan Blue. The decolorization in both systems followed a first-order kinetics. The apparent first-order rate constant, k1', value increases with temperature. Apparent activation energy of decolorization was similar for both systems at ∼ 22 kJ mol(-1), while energy for laccase inactivation was 18 kJ mol(-1). Although both systems were endothermic, fungal biomass showed higher enthalpy, entropy, and Gibbs free energy changes for the decolorization compared to free laccase. On the other hand, free laccase showed reaction spontaneity over a wider range of temperature (ΔT = 40 K) as opposed to fungal biomass (ΔT = 15 K). Comparison of entropy change (ΔS) values indicated metabolism of the dye by the biomass.
Nine aerobic cellulolytic bacterial cultures were obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ) and the American Type Culture Collection (ATCC). The objectives of this study were to characterize the cellulolytic bacteria and to determine the optimum moisture ratio required for solid state fermentation (SSF) of palm kernel cake (PKC). The bacteria cultures were grown on reconstituted nutrient broth, incubated at 30°C and agitated at 200 rpm. Carboxymethyl cellulase, xylanase, and mannanase activities were determined using different substrates and after SSF of PKC. The SSF was conducted for 4 and 7 days with inoculum size of 10% (v/w) on different PKC concentration-to-moisture ratios: 1 : 0.2, 1 : 0.3, 1 : 0.4, and 1 : 0.5. Results showed that Bacillus amyloliquefaciens 1067 DSMZ, Bacillus megaterium 9885 ATCC, Paenibacillus curdlanolyticus 10248 DSMZ, and Paenibacillus polymyxa 842 ATCC produced higher enzyme activities as compared to other bacterial cultures grown on different substrates. The cultures mentioned above also produced higher enzyme activities when they were incubated under SSF using PKC as a substrate in different PKC-to-moisture ratios after 4 days of incubation, indicating that these cellulolytic bacteria can be used to degrade and improve the nutrient quality of PKC.
Toxic inorganic and organic chemicals are major contributors to environmental contamination and pose major health risks to human population. In this work, Dracaena reflexa and Podocarpus polystachyus were investigated for their potential to remove hydrocarbons from 2.5% and 1% diesel fuel-contaminated soil amended individually with 5% organic wastes (tea leaf, soy cake and potato skin) for a period of 270 days. Loss of 90% and 99% oil was recorded in soil contaminated with 2.5% and 1% oil with soy cake amendment, respectively, compared with 52% and 62% in unamended soil with D. reflexa at the end of 270 days. Similarly, 84% and 91% oil loss was recorded for P. polystachyus amended with organic wastes in 2.5% and 1% oil, respectively. Diesel fuel disappeared more rapidly in the soil amendment with SC than in other organic waste supplementation. It was evident that plants did not accumulate hydrocarbon from the soil, while the number of hydrocarbon-utilizing bacteria was high in the rhizosphere, thus suggesting that the mechanism of the oil degradation was rhizodegradation. The kinetic model result indicated a high rate of degradation in soil amendment with SC at 1% with D. reflexa compared with other treatments. Thus, a positive relationship was observed between diesel hydrocarbon degradation with plant biomass production. Dracaena reflexa with organic wastes amendment has a greater potential of restoring hydrocarbon-contaminated soil compared to P. polystachyus plant.
Major degraders of petroleum hydrocarbons in tropical seas have been indicated only by laboratory culturing and never through observing the bacterial community structure in actual environments. To demonstrate the major degraders of petroleum hydrocarbons spilt in actual tropical seas, indigenous bacterial community in seawater at Sentosa (close to a port) and East Coast Park (far from a port) in Singapore was analyzed. Bacterial species was more diverse at Sentosa than at the Park, and the composition was different: γ-Proteobacteria (57.3%) dominated at Sentosa, while they did not at the Park. Specialized hydrocarbonoclastic bacteria (SHCB), which use limited carbon sources with a preference for petroleum hydrocarbons, were found as abundant species at Sentosa, indicating petroleum contamination. On the other hand, SHCB were not the abundant species at the Park. The abundant species of SHCB at Sentosa were Oleibacter marinus and Alcanivorax species (strain 2A75 type), which have previously been indicated by laboratory culturing as important petroleum-aliphatic-hydrocarbon degraders in tropical seas. Together with the fact that SHCB have been identified as major degraders of petroleum hydrocarbons in marine environments, these results demonstrate that the O. marinus and Alcanivorax species (strain 2A75 type) would be major degraders of petroleum aliphatic hydrocarbons spilt in actual tropical seas.
A diverse surfactant, including the nonionic Tween 80 and Brij 30, the anionic sodium dodecyl sulphate, the cationic surfactant Tetradecyltrimethylammonium bromide, and biosurfactant Rhamnolipid were investigated under fluorine-enriched medium by Armilaria sp. F022. The cultures were performed at 25 °C in malt extract medium containing 1 % of surfactant and 5 mg/L of fluorene. The results showed among the tested surfactants, Tween-80 harvested the highest cell density and obtained the maximum specific growth rate. This due Tween-80 provide a suitable carbon source for fungi. Fluorane was also successfully eliminated (>95 %) from the cultures within 30 days in all flasks. During the experiment, laccase production was the highest among other enzymes and Armillaria sp. F022-enriched culture containing Non-ionic Tween 80 showed a significant result for laccase activity (1,945 U/L). The increased enzyme activity was resulted by the increased biodegradation activity as results of the addition of suitable surfactants. The biotransformation of fluorene was accelerated by Tween 80 at the concentration level of 10 mg/L. Fluorene was initially oxidized at C-2,3 positions resulting 9-fluorenone. Through oxidative decarboxylation, 9-fluorenone subjected to meta-cleavage to form salicylic acid. One metabolite detected in the end of experiment, was identified as catechol. Armillaria sp. F022 evidently posses efficient, high effective degrader and potential for further application on the enhanced bioremediation technologies for treating fluorene-contaminated soil.
Phytoremediation is a technology to clean the environment from heavy metals contamination. The objectives of this study are to threat Pb contaminated wastewater by using phytoremediation technology and to determine if the plant can be mention as hyperaccumulator. Fifty plants of Scirpus grossus were grown in sand medium and 600 L spiked water in various Pb concentration (10, 30 and 50 mg/L) was exposed. The experiment was conducted with single exposure method, sampling time on day-1, day-14, day-28, day-42, day-70, and day-98. The analysis of Pb concentration in water, sand medium and inside the plant tissue was conducted by ICP-OES. Water samples were filtered and Pb concentration were directly analyzed, Pb in sand samples were extracted by EDTA method before analyzed, and Pb in plant tissues were extracted by wet digestion method and analyzed. The results showed that on day-28, Pb concentration in water decreased 100%, 99.9%, 99.7%, and the highest Pb uptake by plant were 1343, 4909, 3236 mg/kg for the treatment of 10, 30, and 50 mg/L respectively. The highest BC and TF were 485,261 on day-42 and 2.5295 on day-70 of treatment 30 mg/L, it can be mentioned that Scirpus grossus is a hyperaccumulator.
Polyhydroxyalkanoates (PHAs) are hydrophobic biodegradable thermoplastics that have received considerable attention in biomedical applications due to their biocompatibility, mechanical properties, and biodegradability. In this study, the degradation rate was regulated by optimizing the interaction of parameters that influence the enzymatic degradation of P(3HB) film using response surface methodology (RSM). The RSM model was experimentally validated yielding a maximum 21 % weight loss, which represents onefold increment in percentage weight loss in comparison with the conventional method. By using the optimized condition, the enzymatic degradation by an extracellular PHA depolymerase from Acidovorax sp. DP5 was studied at 37 °C and pH 9.0 on different types of PHA films with various monomer compositions. Surface modification of scaffold was employed using enzymatic technique to create highly porous scaffold with a large surface to volume ratio, which makes them attractive as potential tissue scaffold in biomedical field. Scanning electron microscopy revealed that the surface of salt-leached films was more porous compared with the solvent-cast films, and hence, increased the degradation rate of salt-leached films. Apparently, enzymatic degradation behaviors of PHA films were determined by several factors such as monomer composition, crystallinity, molecular weight, porosity, and roughness of the surface. The hydrophilicity and water uptake of degraded salt-leached film of P(3HB-co-70%4HB) were enhanced by incorporating chitosan or alginate. Salt-leached technique followed by partial enzymatic degradation would enhance the cell attachment and suitable for biomedical as a scaffold.
A white-rot fungus of Polyporus sp. S133 was isolated from an oil-polluted soil. The metabolism of pyrene by this fungus was investigated in liquid medium with 5 mg of the compound. Depletion of pyrene was evident during the 30-day growth period and was 21% and 90%, respectively, in cometabolism and metabolism of pyrene alone. Pyrene was absorbed to fungal cells or biodegraded to form simpler structural compounds. Seventy-one percent of eliminated pyrene was transformed by Polyporus sp. S133 into other compounds, whereas only 18% was absorbed in the fungal cell. The effects of pH and temperature on biomass production of Polyporus sp. S133 for pyrene were examined; the properties of laccase and 1,2-dioxygenase produced by Polyporus sp. S133 during pyrene degradation were investigated. The optimal values of pH were 3, 5, and 4 for laccase, 1,2-dioxygenase, and biomass production, respectively, whereas the optimal values of temperature were 25 °C for laccase and 50 °C for 1,2-dioxygenase and biomass production. Under optimal conditions, pyrene was mainly metabolized to 1-hydroxypyrene and gentisic acid. The structure of 1-hydroxypyrene and gentisic acid was determined by gas chromatography-mass spectrometry after identification using thin-layer chromatography.
In this study, bulrush (Scirpus grossus) was subjected to a 72 day phytotoxicity test to assess its ability to phytoremediate diesel contamination in simulated wastewater at different concentrations (0, 8700, 17,400 and 26,100mg/L). Diesel degradation by S. grossus was measured in terms of total petroleum hydrocarbon (TPH-D). The TPH-D concentration in the synthetic wastewater was determined with the liquid-liquid extraction method and gas chromatography. S. grossus was found to reduce TPH-D by 70.0 and 80.2% for concentrations of 8700 mg/L and 17,400mg/L, respectively. At a diesel concentration of 26,100mg/L, S. grossus died after 14 days. Additionally, the biomass of S. grossus plants was found to increase throughout the phytotoxicity test, confirming the ability of the plant to survive in water contaminated with diesel at rates of less than 17,400mg/L.
A kinetic model incorporating adsorption, desorption and biodegradation processes was developed to describe the bioregeneration of granular activated carbon (GAC) loaded with 4-chlorophenol (4-CP) and 2,4-dichlorophenol (2,4-DCP), respectively, in simultaneous adsorption and biodegradation processes. The model was numerically solved and the results showed that the kinetic model was well-fitted (R(2)>0.83) to the experimental data at different GAC dosages and at various initial 4-CP and 2,4-DCP concentrations. The rate of bioregeneration in simultaneous adsorption and biodegradation processes was influenced by the ratio of initial chlorophenol concentration to GAC dosage. Enhancement in the rate of bioregeneration was achieved by using the lowest ratio under either one of the following experimental conditions: (1) increasing initial chlorophenol concentration at constant GAC dosage and (2) increasing GAC dosage at constant initial chlorophenol concentration. It was found that the rate enhancement was more pronounced under the second experimental condition.
Water hyacinth (Eichhornia crassipes) and water lettuce (Pistia stratiotes) were analyzed to determine their effectiveness in aquaculture wastewater treatment in Malaysia. Wastewater from fish farm in Semanggol Perak, Malaysia was sampled and the parameters determined included, the pH, turbidity, dissolved oxygen (DO), chemical oxygen demand (COD), biochemical oxygen demand (BOD), nitrite phosphate (PO4(3-)), nitrate (NO(3-)), nitrite (NO(-2)), ammonia (NH3), and total kjedahl nitrogen (TKN). Also, hydroponics system was set up and was added with fresh plants weights of 150 +/- 20 grams Eichhornia crassipes and 50 +/- 10 grams Pistia stratiotes during the 30 days experiment. The phytoremediation treatment with Eichhornia crassipes had pH ranging from 5.52 to 5.59 and from 4.45 to 5.5 while Pistia stratiotes had its pH value from 5.76 to 6.49 and from 6.24 to 7.07. Considerable percentage reduction was observed in all the parameters treated with the phytoremediators. Percentage reduction of turbidity for Eichhornia crassipes were 85.26% and 87.05% while Pistia stratiotes were 92.70% and 93.69% respectively. Similar reductions were observed in COD, TKN, NO(3-), NH3, and PO4(3-). The capability of these plants in removing nutrients was established from the study. Removal of aquatic macrophytes from water bodies is recommended for efficient water purification.
Armillaria sp. F022, a white rot fungus isolated from tropical rain forest (Samarinda, Indonesia) was used to biodegrade naphthalene in cultured medium. Transformation of naphthalene by Armillaria sp. F022 which is able to use naphthalene, a two ring-polycyclic aromatic hydrocarbon (PAH) as a source of carbon and energy was investigated. The metabolic pathway was elucidated by identifying metabolites, biotransformation studies and monitoring enzyme activities in cell-free extracts. The identification of metabolites suggests that Armillaria sp. F022 initiates its attack on naphthalene by dioxygenation at its C-1 and C-4 positions to give 1,4-naphthoquinone. The intermediate 2-hydroxybenzaldehyde and salicylic acid, and the characteristic of the meta-cleavage of the resulting diol were identified in the long-term incubation. A part from typical metabolites of naphthalene degradation known from mesophiles, benzoic acid was identified as the next intermediate for the naphthalene pathway of this Armillaria sp. F022. Neither phthalic acid, catechol and cis,cis-muconic acid metabolites were detected in culture extracts. Several enzymes (manganese peroxidase, lignin peroxidase, laccase, 1,2-dioxygenase and 2,3-dioxygenase) produced by Armillaria sp. F022 were detected during the incubation.
Armillaria sp. F022 is a white-rot fungus isolated from a tropical rain forest in Indonesia that is capable of utilizing pyrene as a source of carbon and energy. Enzymes production during the degradation process by Armillaria sp. F022 was certainly related to the increase in biomass. In the first week after incubation, the growth rate rapidly increased, but enzyme production decreased. After 7 days of incubation, rapid growth was observed, whereas, the enzymes were produced only after a good amount of biomass was generated. About 63 % of pyrene underwent biodegradation when incubated with this fungus in a liquid medium on a rotary shaker (120 rpm, 25 °C) for 30 days; during this period, pyrene was transformed to five stable metabolic products. These metabolites were extracted in ethyl acetate, isolated by column chromatography, and then identified using thin layer chromatography (TLC) and gas chromatography-mass spectrometry (GC-MS). 1-Hydroxypyrene was directly identified by GC-MS, while 4-phenanthroic acid, 1-hydroxy-2-naphthoic acid, phthalic acid, and protocatechuic acid were identified to be present in their derivatized forms (methylated forms and silylated forms). Protocatechuic acid was the end product of pyrene degradation by Armillaria sp. F022. Dynamic profiles of two key enzymes, namely laccase and 1,2-dioxygenase, were revealed during the degradation process, and the results indicated the presence of a complicated mechanism in the regulation of pyrene-degrading enzymes. In conclusion, Armillaria sp. F022 is a white-rot fungus with potential for application in the degradation of polycyclic aromatic hydrocarbons such as pyrene in the environment.
The biodegradation of benzo[a]pyrene (BaP) by using Polyporus sp. S133, a white-rot fungus isolated from oil-contaminated soil was investigated. Approximately 73% of the initial concentration of BaP was degraded within 30 d of incubation. The isolation and characterization of 3 metabolites by thin layer chromatography, column chromatography, and UV-vis spectrophotometry in combination with gas chromatography-mass spectrometry, indicated that Polyporus sp. S133 transformed BaP to BaP-1,6-quinone. This quinone was further degraded in 2 ways. First, BaP-1,6-quinone was decarboxylated and oxidized to form coumarin, which was then hydroxylated to hydroxycoumarin, and finally to hydroxyphenyl acetic acid by addition of an epoxide group. Second, Polyporus sp. S133 converted BaP-1,6-quinone into a major product, 1-hydroxy-2-naphthoic acid. During degradation, free extracellular laccase was detected with reduced activity of lignin peroxidase, manganese-dependent peroxidase and 2,3-dioxygenase, suggesting that laccase and 1,2-dioxygenase might play an important role in the transformation of PAHs compounds.