Morphological growth pattern of Phanerochaete chrysosporium cultivated on different Miscanthus x giganteus biomass fractions.

BACKGROUND: Solid-state fermentation is a fungal culture technique used to produce compounds and products of industrial interest. The growth behaviour of filamentous fungi on solid media is challenging to study due to the intermixity of the substrate and the growing organism. Several strategies are available to measure indirectly the fungal biomass during the fermentation such as following the biochemical production of mycelium-specific components or microscopic observation. The microscopic observation of the development of the mycelium, on lignocellulosic substrate, has not been reported. In this study, we set up an experimental protocol based on microscopy and image processing through which we investigated the growth pattern of Phanerochaete chrysosporium on different Miscanthus x giganteus biomass fractions. RESULTS: Object coalescence, the occupied surface area, and radial expansion of the colony were measured in time. The substrate was sterilized by autoclaving, which could be considered a type of pre-treatment. The fastest growth rate was measured on the unfractionated biomass, followed by the soluble fraction of the biomass, then the residual solid fractions. The growth rate on the different fractions of the substrate was additive, suggesting that both the solid and soluble fractions were used by the fungus. Based on the FTIR analysis, there were differences in composition between the solid and soluble fractions of the substrate, but the main components for growth were always present. We propose using this novel method for measuring the very initial fungal growth by following the variation of the number of objects over time. Once growth is established, the growth can be followed by measurement of the occupied surface by the mycelium. CONCLUSION: Our data showed that the growth was affected from the very beginning by the nature of the substrate. The most extensive colonization of the surface was observed with the unfractionated substrate containing both soluble and solid components. The methodology was practical and may be applied to investigate the growth of other fungi, including the influence of environmental parameters on the fungal growth.

The white-rot fungus, Phanerochaete chrysosporium, under combinatorial stress produces variable oil profiles following analysis of secondary metabolites.

AIMS: The effects of combinatorial stress on lipid production in Phanerochaete chrysosporium remain understudied. This species of white-rot fungi was cultivated on solid-state media while under variable levels of known abiotic and biotic stressors to establish the effect upon fungal oil profiles. METHODS AND RESULTS: Environmental stressors induced upon the fungus included the following: temperature, nutrient limitation and interspecies competition to assess impact upon oil profiles. Fatty acid type and its concentration were determined using analytical methods of gas chromatography and mass spectrometry. Growth rate under stress was established using high-performance liquid chromatography with ergosterol as the biomarker. Fungi grown on solid-state agar were able to simultaneously produce short- and long-chain fatty acids which appeared to be influenced by nutritional composition as well as temperature. Addition of nitrogen supplements increased the growth rate, but lipid dynamics remained unchanged. Introducing competition-induced stress had significantly altered the production of certain fatty acids beyond that of the monoculture while under nutrient-limiting conditions. Linoleic acid concentrations, for example, increased from an average of 885 ng µl-1 at monoculture towards 13 820 ng µl-1 at co-culture, following 7 days of incubation. CONCLUSIONS: Interspecies competition produced the most notable impact on lipid production for solid-state media cultivated fungi while the addition of nitrogen supplementation presented growth and lipid accumulation to be uncorrelated. Combinatorial stress therefore influences the yield of overall lipid production as well as the number of intermediate fatty acids produced, deriving similar oil profiles to the composition of vegetable and fish oils. SIGNIFICANCE AND IMPACT OF THE STUDY: Fungal secondary metabolism remains highly sensitive following combinatorial stress. The outcome impacts the research towards optimizing fungal oil profiles for biomass and nutrition. Future investigations on fungal stress tolerance mechanisms need to address these environmental factors throughout the experimental design.

Biofilm Eradication Using Biogenic Silver Nanoparticles.

Microorganisms offer an alternative green and scalable technology for the synthesis of value added products. Fungi secrete high quantities of bioactive substances, which play dual-functional roles as both reducing and stabilizing agents in the synthesis of colloidal metal nanoparticles such as silver nanoparticles, which display potent antimicrobial properties that can be harnessed for a number of industrial applications. The aim of this work was the production of silver nanoparticles using the extracellular cell free extracts of Phanerochaete chrysosporium, and to evaluate their activity as antimicrobial and antibiofilm agents. The 45-nm diameter silver nanoparticles synthesized using this methodology possessed a high negative surface charge close to -30 mV and showed colloidal stability from pH 3-9 and under conditions of high ionic strength ([NaCl] = 10-500 mM). A combination of environmental SEM, TEM, and confocal Raman microscopy was used to study the nanoparticle-E. coli interactions to gain a first insight into their antimicrobial mechanisms. Raman data demonstrate a significant decrease in the fatty acid content of E. coli cells, which suggests a loss of the cell membrane integrity after exposure to the PchNPs, which is also commensurate with ESEM and TEM images. Additionally, these biogenic PchNPs displayed biofilm disruption activity for the eradication of E. coli and C. albicans biofilms.

Natural trypanocidal product produced by endophytic fungi through co-culturing.

Endophytic fungi live inside vegetal tissues without causing damage to the host plant and may provide lead compounds for drug discovery. The co-culture of two or more endophytic fungi can trigger silent gene clusters, which could lead to the isolation of bioactive compounds. In this study, two endophytic strains isolated from Handroanthus impetiginosus leaves, identified as Talaromyces purpurogenus H4 and Phanerochaete sp. H2, were grown in mixed and axenic cultures. The meroterpenoid austin was detected only in the extracts from the mixed culture. Once isolated, austin displayed very interesting trypanocidal activity, with an IC50 value of 36.6 ± 1.2 µg/mL against Trypanosoma cruzi in the epimastigote form. The results obtained highlight the importance of the co-culturing of endophytic fungi to obtain natural bioactive products. The findings also enhance our understanding of the ecological relationships between endophytic fungi.

Changes of heavy metal fractions during co-composting of agricultural waste and river sediment with inoculation of Phanerochaete chrysosporium.

The effects of Phanerochaete chrysosporium on the bioavailability of multiple heavy metals (Pb, Cd, Cu, and Zn) in river sediments were investigated by co-composting with the agricultural waste. The results showed that the Phanerochaete chrysosporium inoculation can greatly enhance the passivation on Cu, Pb and Cd during 60 days co-composting. The effects in the three metals followed the order: Cu > Cd > Pb. There were no differences for Zn whether inoculation with P. chrysosporium or not. Redundancy analysis (RDA) implied that more than 4/5 of the variation of all fractions data for all heavy metals was explained by all significant canonical axes. P. chrysosporium can change the significant parameters for each metal and enhance the explanatory power of RDA model. The inoculation can strengthen the effect of OM (organic matter) on the bioavailability of heavy metals, but weaken the contribution of pH.

Biochemical Characterization of CYP505D6, a Self-Sufficient Cytochrome P450 from the White-Rot Fungus Phanerochaete chrysosporium.

The activity of a self-sufficient cytochrome P450 enzyme, CYP505D6, from the lignin-degrading basidiomycete Phanerochaete chrysosporium was characterized. Recombinant CYP505D6 was produced in Escherichia coli and purified. In the presence of NADPH, CYP505D6 used a series of saturated fatty alcohols with C9-18 carbon chain lengths as the substrates. Hydroxylation occurred at the ω-1 to ω-6 positions of such substrates with C9-15 carbon chain lengths, except for 1-dodecanol, which was hydroxylated at the ω-1 to ω-7 positions. Fatty acids were also substrates of CYP505D6. Based on the sequence alignment, the corresponding amino acid of Tyr51, which is located at the entrance to the active-site pocket in CYP102A1, was Val51 in CYP505D6. To understand the diverse hydroxylation mechanism, wild-type CYP505D6 and its V51Y variant and wild-type CYP102A1 and its Y51V variant were generated, and the products of their reaction with dodecanoic acid were analyzed. Compared with wild-type CYP505D6, its V51Y variant generated few products hydroxylated at the ω-4 to ω-6 positions. The products generated by wild-type CYP102A1 were hydroxylated at the ω-1 to ω-4 positions, whereas its Y51V variant generated ω-1 to ω-7 hydroxydodecanoic acids. These observations indicated that Val51 plays an important role in determining the regiospecificity of fatty acid hydroxylation, at least that at the ω-4 to ω-6 positions. Aromatic compounds, such as naphthalene and 1-naphthol, were also hydroxylated by CYP505D6. These findings highlight a unique broad substrate spectrum of CYP505D6, rendering it an attractive candidate enzyme for the biotechnological industry.IMPORTANCEPhanerochaete chrysosporium is a white-rot fungus whose metabolism of lignin, aromatic pollutants, and lipids has been most extensively studied. This fungus harbors 154 cytochrome P450-encoding genes in the genome. As evidenced in this study, P. chrysosporium CYP505D6, a fused protein of P450 and its reductase, hydroxylates fatty alcohols (C9-15) and fatty acids (C9-15) at the ω-1 to ω-7 or ω-1 to ω-6 positions, respectively. Naphthalene and 1-naphthol were also hydroxylated, indicating that the substrate specificity of CYP505D6 is broader than those of the known fused proteins CYP102A1 and CYP505A1. The substrate versatility of CYP505D6 makes this enzyme an attractive candidate for biotechnological applications.

Antioxidative response of Phanerochaete chrysosporium against silver nanoparticle-induced toxicity and its potential mechanism.

Antioxidative response of Phanerochaete chrysosporium induced by silver nanoparticles (AgNPs) and their toxicity mechanisms were comprehensively investigated in a complex system with 2,4-dichlorophenol (2,4-DCP) and Ag+. Malondialdehyde content was elevated by 2,4-DCP, AgNPs, and/or Ag+ in concentration- and time-dependent manners within 24 h, indicating an increase in lipid peroxidation. However, beyond 48 h of exposure, lipid peroxidation was alleviated by upregulation of intracellular protein production and enhancement in the activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD). Comparatively, POD played more major roles in cell protection against oxidative damage. Furthermore, the dynamic change in reactive oxygen species (ROS) level was parallel to that of oxidized glutathione (GSSG), and ROS levels correlated well with GSSG contents (R2 = 0.953) after exposure to AgNPs for 24 h. This finding suggested that elimination of oxidative stress resulted in depletion of reduced glutathione. Coupled with the analyses of anoxidative responses of P. chrysosporium under the single and combined treatments of AgNPs and Ag+, HAADF-STEM, SEM, and EDX demonstrated that AgNP-induced cytotoxicity could originate from the original AgNPs, rather than dissolved Ag+ or the biosynthesized AgNPs.

Lignin peroxidase ligand access channel dysfunction in the presence of atrazine.

Studies have determined that the white-rot basidiomycete Phanerochaete chrysosporium is capable of biodegrading the atrazine herbicide with its broad-specificity enzymes, but the particular role of biocatalysts is still unclear. In the case of lignin peroxidase, a ligand access channel connected to the active heme cofactor provides access to the active site for potential small-sized substrates. Experimental results show that lignin peroxidase is unable to degrade atrazine, therefore, the primary goal was to determine whether there is any connection between the structural and dynamical properties of the enzyme and its incapability to degrade atrazine. The results of protein-ligand docking and molecular dynamics study correlate with relevant, published NMR and molecular dynamics data, and give the answer to the lack of atrazine degradation by lignin peroxidase which has already been established by numerous authors using experimental methods. Atrazine has no access to heme edge due to the electric charges of the delocalized s-triazine ring. The detected phenomenon suggests that the small size of the ligands only is not a sufficient condition to access the active site. Their physicochemical properties influence the structural behaviour of the channel.

Use of non-living lyophilized Phanerochaete chrysosporium cultivated in various media for phenol removal.

The biosorption of phenol on non-living lyophilized mycelial pellets of Phanerochaete chrysosporium cultivated in liquid medium of various compositions was studied in batch biosorption system. The fungal cell surfaces were characterized by FTIR spectroscopy and specific surface charge determination. The sorption kinetics and equilibrium were evaluated using linear and non-linear regression. For adsorption equilibrium, a comparative evaluation is also presented using non-linear least-square estimation and linearization of the Langmuir and anti-Langmuir equations. The presence of mineral and vitamin materials in the liquid medium enhanced the adsorption capacity of fungal biomass for phenol. At optimum pH 5-6, the values of specific surface charge were 0.023 and 0.069 meq g-1 for various cultivations, and the maximum amounts of phenol can be adsorbed at these pH values. The maximum adsorbed phenol amounts by cells cultivated in simple and complex media were 4.53 and 13.48 mg g-1, respectively, at an initial phenol concentration of 100 mg l-1. Graphical abstract ᅟ.

White rot fungi and advanced combined biotechnology with nanomaterials: promising tools for endocrine-disrupting compounds biotransformation.

Endocrine-disrupting compounds (EDCs) can interfere with endocrine systems and bio-accumulate through the food chain and even decrease biodiversity in contaminated areas. This review discusses a critical overview of recent research progress in the biotransformation of EDCs (including polychlorinated biphenyl and nonylphenol, and suspected EDCs such as heavy metals and sulfonamide antibiotics) by white rot fungi (WRF) based on techniques with an emphasis on summarizing and analyzing fungal molecular, metabolic and genetic mechanisms. Not only intracellular metabolism which seems to perform essential roles in the ability of WRF to transform EDCs, but also advanced applications are deeply discussed. This review mainly reveals the removal pathway of heavy metal and antibiotic pollutants because the single pollution almost did not exist in a real environment while the combined pollution has become more serious and close to people's life. The trends in WRF technology and its related advanced applications which use the combined technology, including biocatalysis of WRF and adsorption of nanomaterials, to degrade EDCs have also been introduced. Furthermore, challenges and future research needs EDCs biotransformation by WRF are also discussed. This research, referring to metabolic mechanisms and the combined technology of WRF with nanomaterials, undoubtedly contributes to the applications of biotechnology. This review will be of great benefit to an understanding of the trends in biotechnology for the removal of EDCs.

Bioaccumulation and toxicity of CdSe/ZnS quantum dots in Phanerochaete chrysosporium.

The growing potential of quantum dots (QDs) in biomedical applications has raised considerable concerns regarding their toxicological impact. Consequently, there has been a need to understand the underlying toxicity mechanism of QDs. In this work, we comprehensively investigated the bioaccumulation and toxicity of three CdSe/ZnS QDs (COOH CdSe/ZnS 525, NH2 CdSe/ZnS 525, and NH2 CdSe/ZnS 625) in Phanerochaete chrysosporium (P. chrysosporium) using confocal laser scanning microscopy, reactive oxygen species (ROS) measurements, and cell viability assays. Confocal laser scanning microscopy analytical results indicated that all the CdSe/ZnS QDs, with the concentration ranging from 10 to 80nM, could accumulate largely in the hyphae and induce the generation of ROS, showing a direct toxicity to P. chrysosporium. And the bioaccumulation and toxicity of CdSe/ZnS QDs presented dose-dependent and time-dependent effects on P. chrysosporium. Furthermore, the CdSe/ZnS QDs-induced cytotoxicity was also related to their physicochemical properties, including particle size and surface charges: NH2 CdSe/ZnS 525 with small size was more cytotoxic as compared to NH2 CdSe/ZnS 625 with large size, and the smaller negative charged NH2 CdSe/ZnS 525 resulted in greater cytotoxicity than the larger negative charged COOH CdSe/ZnS 525. The obtained results provide valuable information for exploring and understanding of toxicity mechanism of QDs in living cells.

Organic matters removal from landfill leachate by immobilized Phanerochaete chrysosporium loaded with graphitic carbon nitride under visible light irradiation.

This study investigated the technical applicability of a combination of Phanerochaete chrysosporium (P. chrysosporium) with photocatalyst graphitic carbon nitride (g-C3N4) for organic matters removal from landfill leachate under visible light irradiation. Photocatalyst g-C3N4 was well immobilized on the hyphae surface of P. chrysosporium by calcium alginate. The typical absorption edge in visible light region for g-C3N4 was at about 460 nm, and the optical absorption bandgap of g-C3N4 was estimated to be 2.70 eV, demonstrating the great photoresponsive ability of g-C3N4. An optimized g-C3N4 content of 0.10 g in immobilized P. chrysosporium and an optimized immobilized P. chrysosporium dosage of 1.0 g were suitable for organic matters removal. The removal efficiency of total organic carbon (TOC) reached 74.99% in 72 h with the initial TOC concentration of 100 mg L-1. In addition, the gas chromatography coupled with mass spectrometry (GC-MS) measurements showed that immobilized P. chrysosporium presented an outstanding removal performance for almost all organic compounds in landfill leachate, especially for the volatile fatty acids and long-chain hydrocarbons. The overall results indicate that the combination P. chrysosporium with photocatalyst g-C3N4 for organic matters removal from landfill leachate may provide a more comprehensive potential for the landfill leachate treatment.

Crystal structure of a family 6 cellobiohydrolase from the basidiomycete Phanerochaete chrysosporium.

Cellobiohydrolases belonging to glycoside hydrolase family 6 (CBH II, Cel6A) play key roles in the hydrolysis of crystalline cellulose. CBH II from the white-rot fungus Phanerochaete chrysosporium (PcCel6A) consists of a catalytic domain (CD) and a carbohydrate-binding module connected by a linker peptide, like other known fungal cellobiohydrolases. In the present study, the CD of PcCel6A was crystallized without ligands, and p-nitrophenyl ß-D-cellotrioside (pNPG3) was soaked into the crystals. The determined structures of the ligand-free and pNPG3-soaked crystals revealed that binding of cellobiose at substrate subsites +1 and +2 induces a conformational change of the N-terminal and C-terminal loops, switching the tunnel-shaped active site from the open to the closed form.

Biosorption of landfill leachate by Phanerochaete sp. ISTL01: isotherms, kinetics and toxicological assessment.

The study investigates the ability of fungus Phanerochaete sp. ISTL01 for biosorption of color from landfill leachate. Batch mode experiments were conducted to study the effects of pH, temperature, adsorbent dose, contact time and initial leachate concentration on biosorption. Maximum biosorption capacity was determined as 17.73 mg g-1 of biomass. Equilibrium isotherms and kinetics were further studied. The biosorption data were found to fit well to the Freundlich isotherm and pseudo-second-order kinetic model. The value of activation energy suggested that chemisorption mechanism was involved. Biosorption efficiency was also evaluated by the Methyltetrazolium (MTT) assay for cytotoxicity and alkaline comet assay in HepG2 human hepato-carcinoma cells. The fungus reduced toxicity as shown by 1.3-fold increase in MTT EC50 and 1.5- and 1.1-fold reduction in Tail moment and Olive tail moment, respectively, after 12 h biosorption. The fungus showed good biosorption characteristics in terms of contaminant-level reduction per unit mass of adsorbent, process kinetics and toxicity reduction, envisaging its application in leachate treatment.

Sorption of zinc onto elemental selenium nanoparticles immobilized in Phanerochaete chrysosporium pellets.

The use of a novel hybrid biosorbent, elemental selenium nanoparticles (nSe0) immobilized in pellets of Phanerochaete chrysosporium, to remove Zn from aqueous solutions was investigated. Fungal pellets containing nSe0 (nSe0-pellets) showed to be better biosorbents as they removed more Zn (88.1 ± 5.3 %) compared to Se-free fungal pellets (56.2 ± 2.8 %) at pH 4.5 and an initial Zn concentration of 10 mg L-1. The enhanced sorption capacity of nSe0-pellets was attributed to a higher concentration of sorption sites resulting in a more negative surface charge density, as determined by analysis of the potentiometric titration data. Fourier transform infrared spectroscopy (FT-IR) analysis of fungal pellets prior to and after being loaded with Zn showed the functional groups, including hydroxyl and carboxyl groups, involved in the sorption process. The experimental data indicated that the sorption rate of the nSe0-pellets fitted well to the pseudo-second order kinetic model (R 2  = 0.99), and the sorption isotherm was best represented by the Sips model (Langmuir-Freundlich) with heterogeneous factor n = 1 (R 2  = 0.99), which is equivalent to the Langmuir model. Operational advantages of fungal pelleted reactors and the Zn removal efficiencies achieved by nSe0-pellets under mild acidic conditions make nSe0-pellet based bioreactors an efficient biosorption process.

Chitosan beads immobilized manganese peroxidase catalytic potential for detoxification and decolorization of textile effluent.

Textile industry has led to severe environmental pollution and is posing a serious threat to the ecosystems. Immobilized biocatalysts have gained importance as potential bio-remediating agent. Manganese peroxidase (MnP) was immobilized onto glutaraldehyde activated chitosan beads by crosslinking and employed for the degradation and detoxification of dyes in textile effluents. The efficiency of chitosan-immobilized MnP (CI-MnP) was evaluated on the basis of decolorization, water quality improvement and toxicity reduction. Maximum color removal of 97.31% was recorded and up to 82.40%, 78.30% and 91.7% reductions in COD, TOC, and BOD were achieved, respectively. The cytotoxicity of bio-treated effluents reduced significantly and 38.46%, 43.47% and 41.83% Allium cepa root length, root count and mitotic index were increased, respectively, whereas brine shrimp nauplii death reduced up to 63.64%. Mutagenicity (Ames test) reduced up to 73.44% and 75.43% for TA98 and TA100 strains, respectively. The CI-MnP retained 60% activity after 10 repeated decolorization batches. The CI-MnP showed excellent efficiency for the bioremediation of textile effluents and can be used for the remediation of toxic agents in wastewater. The monitoring of processed wastewater using bioassays is suggested to evaluate bio-efficiency of treatment method for safe disposal of effluents into water bodies.

Removal of phenol in phenolic resin wastewater by a novel biomaterial: the Phanerochaete chrysosporium pellet containing chlamydospore-like cells.

A novel biomaterial, the Phanerochaete chrysosporium pellet (CP) composed of chlamydospore-like cells (CLCs), was prepared and its potential in treating phenolic resin wastewater was evaluated. CP possesses higher phenol removal ability in contrast with mycelial pellets of P. chrysosporium, and CLC can be seen as the naturally immobilized enzymes. At shake-flask level, the ideal pH value, temperature, and inoculation quantity of CP for treatment of 1430 mg/l phenol wastewater were pH 4-6, 30 °C, and 5.0 g/l, respectively, and the maximum specific removal rate, 41.1 mg phenol/g CP/h, was obtained in fixed bed reactor (FBR) when the flow rate of wastewater was 3.4 l/h. During the treatment, FBR harbored amounts of bacteria (135 genera) and eukaryotes, as analyzed by metagenomic sequencing. Bacterial pollution not only decreased reactor performance but also had a negative impact on reusability of CP. Hot water treatment (80-85 °C) is effective to inhibit bacterial pollution, and heat resistance of CLC makes the repeated regrowing of CP be feasible. This work presents an innovative and low-cost biomaterial for phenol removal and will be helpful for the practical application of P. chrysosporium in wastewater treatment.

[Comparison of two mycelial pellets formation methods to immobilize o-chlorophenol degradation bacteria].

Objective: To find an efficient and fast method for microbial immobilization, we compared simultaneity culture method and adsorption method on morphology and intrastructure of combined mycelial pellets, as well as their o-chlorophenol biodegradation efficiency. Methods: The o-chlorophenol degrading photosynthetic bacterium PSB-1D was immobilized onto mycelial pellets formed by Phanerochaete chrysosporium DH-1 to form combined mycelial pellets. The morphology and intrastructure of pellets formed by two immobilization methods were observed by optical microscope and scanning electron microscope. Then, their differences were analyzed. Using the sterile medium as control, o-chlorophenol removal efficiency of free photosynthetic bacteria, single mycelial pellets and combined mycelial pellets formed by two methods were studied. Results: Photosynthetic bacteria were largely concentrated in the core region of pellets formed by simultaneity culture method and grew in clusters on each mycelium and their intersections. As compared with simultaneity culture method, photosynthetic bacteria mainly grew in the transition region of pellets formed by adsorption method. With the same inoculation amount of spores and photosynthetic bacteria, the simultaneity culture method could immobilize more bacteria with little time. Moreover, average diameter, dry weight and dry wet ratio of pellets formed by simultaneity culture method were bigger than that by adsorption method, and their desorption amount were less. The o-chlorophenol degradation followed a first-order kinetics model. The combined mycelial pellets formed by simultaneity culture method could degrade above 89% of o-chlorophenol in medium with an initial concentration of 50 mg/L after incubation for 7 days. And the half-life periods (t1/2) were shortened to 2.8 days. Conclusion: The study provides the theoretical foundation for the practical application of the new biomass carrier to organic wastewater treatment.

Treatment of landfill leachate using immobilized Phanerochaete chrysosporium loaded with nitrogen-doped TiO2 nanoparticles.

This study investigated the performance of immobilized Phanerochaete chrysosporium loaded with nitrogen-doped TiO2 nanoparticles in the treatment of raw landfill leachate with a very low biodegradability ratio (BOD5/COD) of 0.09. The effects of various operating parameters, such as initial chemical oxygen demand (COD) concentration, pH, temperature, and biosorbent dosage, were evaluated with respect to the removal efficiency of total organic carbon (TOC) and ammonia nitrogen (NH3-N). For the immobilized biosorbents, an optimum pH of 6.0 for TOC and 7.0 for NH3-N were found suitable for TOC and NH3-N removal at temperature of 37°C, respectively. The most superior removal efficiencies of TOC and NH3-N of landfill leachate were over 75% and 74% in 72 h at an initial COD concentration of 200 mg L(-1), respectively. In addition, heavy metals were partly removed by the immobilized biosorbents during the process of landfill leachate treatment. The species and mass percentage of organic compounds in landfill leachate after the treatment were found to have considerably declined according to the gas chromatography coupled with mass spectrometry (GC-MS) system. These results indicate that the immobilized P. chrysosporium loaded with nitrogen-doped TiO2 nanoparticles could be a convenient and efficient method for the treatment of landfill leachate.

Removal of humic acid from composted hog waste by the white-rot fungus, Phanerochaete chrysosporium.

The potential hazards of humic acid (HA) associated with hog waste effluent, coupled with increasing awareness of environmental problems, have prompted many countries to control disposal of effluents into water bodies and to maximize removal of HA. Here we employed the white-rot fungus, Phanerochaete chrysosporium, to degrade the HA in composted hog waste effluent, evaluated by the response surface method. Preliminary experiments demonstrate that pH, temperature and quantity of inoculum are significant variables determining success of the fungus. In total, 13 experiments were conducted with three variables designated as A (pH), B (temperature) and C (inoculum amount). The optimal conditions for reduction of HA by P. chrysosporium are pH 6, 31.5°C and an inoculum quantity of 5.86 g. Predicted and experimental results exhibit strong agreement, indicating efficiency in the model obtained by response surface method. Therefore, P. chrysosporium is an effective micro-organism for removal of HA from composted hog waste effluent.