Fungal Bioremediation Research Articles

Trichoderma: a potential bioremediator for environmental clean up

Environmental awareness has resulted in development of regulatory measures that aim to straighten past mistakes and protect the environment from future contamination and exploitation. However, much consideration and research needs to go into the decision-making process for an effective clean up of a particular contaminated site. Each technology developed has its advantages and limitations for the treatment of specific contaminants. Bioremediation and phytoremediation in association with microbes are innovative technologies having a potential to alleviate numerous environmental pollution problems. Owing to its dominant presence in contaminated sites, the application of the fungi in bioremediation is well documented. The genus Trichoderma is genetically very diverse with a number of capabilities among different strains with agricultural and industrial significance. It is also tolerant to a range of recalcitrant pollutants including heavy metals, pesticides, and polyaromatic hydrocarbons. This review presents an updated overview of application of Trichoderma for biological or phytobial remediation of environmental contaminants.

Enhanced oxidation of benzo[a]pyrene by crude enzyme extracts produced during interspecific fungal interaction of Trametes versicolor and Phanerochaete chrysosporium

The effects of interspecific fungal interactions between Trametes versicolor and Phanerochaete chrysosporium on laccase activity and enzymatic oxidation of polycyclic aromatic hydrocarbons (PAHs) were investigated. A deadlock between the two mycelia rather than replacement of one fungus by another was observed on an agar medium. The laccase activity in crude enzyme extracts from interaction zones reached a maximum after a 5-day incubation, which was significantly higher than that from regions of T. versicolor or P. chrysosporium alone. The enhanced induction of laccase activity lasted longer in half nutrition than in normal nutrition. A higher potential to oxidize benzo[a]pyrene by a crude enzyme preparation extracted from the interaction zones was demonstrated. After a 48 hr incubation period, the oxidation of benzo[a]pyrene by crude enzyme extracts from interaction zones reached 26.2%, while only 9.5% of benzo[a]pyrene was oxidized by crude extracts from T. versicolor. The oxidation was promoted by the co-oxidant 2,2′-azinobis-3-ethylbenzthiazoline-6-sulphonate diammonium salt (ABTS). These findings indicate that the application of co-culturing of white-rot fungi in bioremediation is a potential ameliorating technique for the restoration of PAH-contaminated soil.

Degradation pathway of pentachlorophenol by Mucor plumbeus involves phase II conjugation and oxidation–reduction reactions

Environmental pollution by pentachlorophenol (PCP) is a critical concern worldwide and fungal bioremediation constitutes an elegant and environment-friendly solution. Mucorales from the Zygomycota phylum are often observed to be competitive in field conditions and Mucor plumbeus, in particular, can efficiently deplete PCP from media. The pathway for PCP degradation used by this fungus has not been investigated. In this study, PCP-derived metabolites were identified by liquid chromatography coupled with quadrupole time-of-flight mass spectrometry, including tetra- and tri-chlorohydroquinones and phase II-conjugated metabolites. Amongst the latter are the previously reported glucose, sulfate and ribose conjugates, and identified for the first time in fungi sulfate–glucose conjugates. A PCP transformation pathway for M. plumbeus is proposed, which excludes the involvement of cytochrome P-450 and extracellular ligninolytic enzymes.

Untapped potential: exploiting fungi in bioremediation of hazardous chemicals

Fungi possess the biochemical and ecological capacity to degrade environmental organic chemicals and to decrease the risk associated with metals, metalloids and radionuclides, either by chemical modification or by influencing chemical bioavailability. Furthermore, the ability of these fungi to form extended mycelial networks, the low specificity of their catabolic enzymes and their independence from using pollutants as a growth substrate make these fungi well suited for bioremediation processes. However, despite dominating the living biomass in soil and being abundant in aqueous systems, fungi have not been exploited for the bioremediation of such environments. In this Review, we describe the metabolic and ecological features that make fungi suited for use in bioremediation and waste treatment processes, and discuss their potential for applications on the basis of these strengths.

Screening and selection of fungi for bioremediation of olive mill wastewater

Olive mill wastewater (OMWW) is a significant pollutant and its phytotoxicity is attributed mostly to the phenols present. 220 fungi were screened for their ability to produce detoxifying enzymes and/or grow in OMWW. Four isolates, species of Cerrena, Byssochlamys, Lasiodiplodia and Bionectria were selected and compared against Phanerochaete chrysosporium for their ability to bioremediate OMWW in the presence of a competing indigenous microflora. For the first time we report that a Cerrena sp. achieved a 75% reduction of phenolics in OMWW and that, unusually, the reduction occurred within 2 h of the addition to the OMWW.

Four marine-derived fungi for bioremediation of raw textile mill effluents

Textile dye effluents pose environmental hazards because of color and toxicity. Bioremediation of these has been widely attempted. However, their widely differing characteristics and high salt contents have required application of different microorganisms and high dilutions. We report here decolorization and detoxification of two raw textile effluents, with extreme variations in their pH and dye composition, used at 20-90% concentrations by each of the four marine-derived fungi. Textile effluent A (TEA) contained an azo dye and had a pH of 8.9 and textile effluent B (TEB) with a pH of 2.5 contained a mixture of eight reactive dyes. The fungi isolated from mangroves and identified by 18S and ITS sequencing corresponded to two ascomycetes and two basidiomycetes. Each of these fungi decolorized TEA by 30-60% and TEB by 33-80% used at 20-90% concentrations and salinity of 15 ppt within 6 days. This was accompanied by two to threefold reduction in toxicity as measured by LC(50) values against Artemia larvae and 70-80% reduction in chemical oxygen demand and total phenolics. Mass spectrometric scan of effluents after fungal treatment revealed degradation of most of the components. The ascomycetes appeared to remove color primarily by adsorption, whereas laccase played a major role in decolorization by basidiomycetes. A process consisting of a combination of sorption by fungal biomass of an ascomycete and biodegradation by laccase from a basidiomycete was used in two separate steps or simultaneously for bioremediation of these two effluents.

Fungal bioremediation of chromates: Conformational changes of biomass during sequestration, binding, and reduction of hexavalent chromium ions

This paper highlights the mechanistic aspects of white rot fungus Coriolus versicolor as a complexing/reducing agent for chromium bioremediation. The chemical reduction of Cr(VI) to Cr(III) via the formation of Cr(VI) thio ester as an intermediate, is pH dependent and controls the overall chromium adsorption kinetics. The strong adsorption affinity of the biomass towards Cr(VI) anions was evaluated by the Freundlich and the Langmuir adsorption isotherms. The FTIR spectroscopic analysis suggested the involvement of amino, carboxylate, and thiol groups from fungal cell wall in chromium binding and reduction. The mechanism of the adsorption was preferential sequestration along with binding of the metal to the ligating groups present in the biomass followed by reduction to trivalent state. The results indicate step-wise progression of overall reaction dictated and modulated by structural and conformation effects in the biomass that lead to saturation, acceleration, and ultimate saturation kinetics effects.

Fungal bioremediation of the creosote-contaminated soil: Influence of Pleurotus ostreatus and Irpex lacteus on polycyclic aromatic hydrocarbons removal and soil microbial community composition in the laboratory-scale study

The aim of this study was to determine the efficacy of selected basidiomycetes in the removing of polycyclic aromatic hydrocarbons (PAH) from the creosote-contaminated soil. Fungi Pleurotus ostreatus and Irpex lacteus were supplemented with creosote-contaminated (50–200 mg kg −1 PAH) soil originating from a wood-preserving plant and incubated at 15 °C for 120 d. Either fungus degraded PAH with 4–6 aromatic rings more efficiently than the microbial community present initially in the soil. PAH removal was higher in P. ostreatus treatments (55–67%) than in I. lacteus treatments (27–36%) in general. P. ostreatus (respectively, I. lacteus) removed 86–96% (47–59%) of 2-rings PAH, 63–72% (33–45%) of 3-rings PAH, 32–49% (9–14%) of 4-rings PAH and 31–38% (11–13%) of 5–6-rings PAH. MIS (Microbial Identification System) Sherlock analysis of the bacterial community determined the presence of dominant Gram-negative bacteria (G−) Pseudomonas in the inoculated soil before the application of fungi. Complex soil microbial community was characterized by phospholipid fatty acids analysis followed by GC–MS/MS. Either fungus induced the decrease of bacterial biomass (G− bacteria in particular), but the soil microbial community was influenced by P. ostreatus in a different way than by I. lacteus. The bacterial community was stressed more by the presence of I. lacteus than P. ostreatus (as proved by the ratio of the fungal/bacterial markers and by the ratio of trans/ cis mono-unsaturated fatty acids). Moreover, P. ostreatus stimulated the growth of Gram-positive bacteria (G+), especially actinobacteria and these results indicate the potential of the positive synergistic interaction of this fungus and actinobacteria in creosote biodegradation.

Fungal Bioremediation of Phenolic Wastewaters in an Airlift Reactor

Of the various types of industry-generated effluents, those containing organic pollutants such as phenols are generally difficult to remediate. There is a need to develop new technologies that emphasize the destruction of these pollutants rather than their disposal. In this work the white rot fungus, Trametes pubescens, was demonstrated to be an effective bioremediation agent for the treatment of phenolic wastewaters. An airlift loop reactor was optimized, in terms of volumetric oxygen transfer rate (K(L)a = 0.45 s(-1)), to provide an environment suited to rapid growth of T.pubescens (mu = 0.25 day(-1)) and a particularly efficient growth yield on glucose of 0.87 g biomass.g glucose(-1). The phenolic effluent was shown to be a paramorphogen, influencing fungal pellet morphology in the reactor, as well as increasing laccase enzyme activity by a factor of 5 over the control, to a maximum of 11.8 U.mL(-1). This increased activity was aided by the feeding of nonrepressing amounts (0.5 g.L(-1)) of glucose to the reactor culture. To our knowledge the degradation results represent the highest rate of removal (0.033 g phenol.g biomass(-1).day(-1)) of phenolic compounds from water reported for white rot fungi.

Pilot Study of Bioaccumulation and Distribution of Cesium, Potassium, Sodium and Calcium in King Oyster Mushroom (Pleurotus Eryngii) Grown Under Controlled Conditions

This pilot study presents preliminary results on interrelations between alkali and alkaline earth elements during their transfer to mycelium and fruitbodies of saprophytic fungi. The accumulation and distribution of four elements (cesium, potassium, sodium, and calcium) was evaluated in king oyster mushroom (Pleurotus eryngii) cultivated under controlled conditions. Elemental composition of caps, stipes, and the substrate was analyzed by atomic absorption/emission spectroscopy to evaluate discrimination, concentration, and transfer factors. The transfer factors determined for all the investigated elements were different and can be put in the following order: Cs > K > Na > Ca. There has been a higher accumulation of cesium in caps than in stipes. Distribution of cesium in fruitbodies depended on the presence of other ions in the substrate. The addition of Ca2+ limited the transport of cesium and potassium from stipes to caps. Sodium and calcium were mainly accumulated in the stipes. In a control experiment, without supplementation with K+, Na+, and Ca2+, ∼ 62% of the cesium present in the substrate was extracted by mycelium and transported to the fruitbodies. Possible applications of fruiting saprophytic fungi in bioremediation are discussed.

Fungi and ionizing radiation from radionuclides

Radionuclides in the environment are one of the major concerns to human health and ecotoxicology. The explosion at the Chernobyl nuclear power plant renewed interest in the role played by fungi in mediating radionuclide movement in ecosystems. As a result of these studies, our knowledge of the importance of fungi, especially in their mycorrhizal habit, in long-term accumulation of radionuclides, transfer up the food chain and regulation of accumulation by their host plants was increased. Micro-fungi have been found to be highly resilient to exposure to ionizing radiation, with fungi having been isolated from within and around the Chernobyl plant. Radioresistance of some fungal species has been linked to the presence of melanin, which has been shown to have emerging properties of acting as an energy transporter for metabolism and has been implicated in enhancing hyphal growth and directed growth of sensitized hyphae towards sources of radiation. Using this recently acquired knowledge, we may be in a better position to suggest the use of fungi in bioremediation of radioactively contaminated sites and cleanup of industrial effluent.

Book Reviews

Book Reviews

The fine‐tuning of heavy metals in mycorrhizal fungi

The fine‐tuning of heavy metals in mycorrhizal fungi

Bioremediation of Hazardous Wastes—A Review

Intensive industrialization generates hazardous wastes comprising organics, inorganics, heavy metals, and munitions that need to be tackled in a safe manner. Commonly employed physicochemical technologies have paved the way to ecofriendly bioremediation processes. Bioremediation uses natural as well as recombinant microorganisms to break down toxic and hazardous substances by aerobic and anaerobic means. They can be applied on site (in situ) or off site (ex situ), mediated by mixed microbial consortia and/or pure microbial strains and. plants (phytoremediation) or even natural attenuation. They include several processes—bioventing, biosparging, biostimulation, bioaugmentation, bioleaching, fungal bioremediation, and biosorption. Bioremediation also encompasses ex situ engineered methods like bioreactors and enzyme catalyzed breakdown. The success of bioremediation is governed by three important factors—availability of microbes, accessibility of contaminants, and a conducive environment. This review discusses various bioremediation technologies, listing the advantages as well as disadvantages and field application, if any.

Bioavailable cadmium during the bioremediation of phenanthrene-contaminated soils using the diffusive gradients in thin-film technique

To study the impact of fungal bioremediation of phenanthrene on trace cadmium solid-solution fluxes and solution phase concentration. The bioremediation of phenanthrene in soils was performed using the fungus Penicillium frequentans. Metal behaviour was evaluated by the techniques of diffusive gradient in thin-films (DGT) and filtration. Fluxes of cadmium (Cd) show a significant (P < 0.002) increase after the start of bioremediation, indicating that the bioremediation process itself releases significant amount of Cd into solution from the soil solid-phase. Unlike DGT devices, the solution concentration from filtration shows a clear bimodal distribution. We postulate that the initial action of the fungi is most likely to breakdown the surface of the solid phase to smaller, 'solution-phase' material (<0.45 microm) leading to a peak in Cd concentration in solution. Phenanthrene removal from soils by bioremediation ironically results in the mobilization of another toxic pollutant (Cd). Bioremediation of organic pollutants in contaminated soil will likely lead to large increases in the mobilization of toxic metals, increasing metal bio-uptake and incorporation into the wider food chain. Bioremediation strategies need to account for this behaviour and further research is required both to understand the generality of this behaviour and the operative mechanisms.

Role of Autochthonous Filamentous Fungi in Bioremediation of a Soil Historically Contaminated with Aromatic Hydrocarbons

Nine fungal strains isolated from an aged and heavily contaminated soil were identified and screened to assess their degradative potential. Among them, Allescheriella sp. strain DABAC 1, Stachybotrys sp. strain DABAC 3, and Phlebia sp. strain DABAC 9 were selected for remediation trials on the basis of Poly R-478 decolorization associated with lignin-modifying enzyme (LME) production. These autochthonous fungi were tested for the abilities to grow under nonsterile conditions and to degrade various aromatic hydrocarbons in the same contaminated soil. After 30 days, fungal colonization was clearly visible and was confirmed by ergosterol determination. In spite of subalkaline pH conditions and the presence of heavy metals, the autochthonous fungi produced laccase and Mn and lignin peroxidases. No LME activities were detected in control microcosms. All of the isolates led to a marked removal of naphthalene, dichloroaniline isomers, o-hydroxybiphenyl, and 1,1'-binaphthalene. Stachybotrys sp. strain DABAC 3 was the most effective isolate due to its ability to partially deplete the predominant contaminants 9,10-anthracenedione and 7H-benz[DE]anthracen-7-one. A release of chloride ions was observed in soil treated with either Allescheriella sp. strain DABAC 1 or Stachybotrys sp. strain DABAC 3, suggesting the occurrence of oxidative dehalogenation. The autochthonous fungi led to a significant decrease in soil toxicity, as assessed by both the Lepidium sativum L. germination test and the Collembola mortality test.

Influence of corn steep liquor and glucose on colonization of control and CCB (Cu/Cr/B)-treated wood by brown rot fungi

There are increasing problems with regard to the disposal of treated wood waste. Due to heavy metals or arsenic in impregnated wood waste, burning and landfill disposal options are not considered to be environmentally friendly solutions for dealing with this problem. Extraction of the heavy metals and recycling of the preservatives from the wood waste is a much more promising and environmentally friendly solution. In order to study the scale up of this process, copper/chromium/boron-treated wood specimens were exposed to copper tolerant ( Antrodia vaillantii and Leucogyrophana pinastri) and copper sensitive wood decay fungi ( Gloeophyllum trabeum and Poria monticola). Afterwards, the ability of fungal hyphae to penetrate and overgrow the wood specimens was investigated. The fungal growths were stimulated by immersing the specimens into aqueous solution of glucose or corn steep liquor prior to exposure to the fungi. The fastest colonization of the impregnated wood was by the copper tolerant A. vaillantii. Addition of glucose onto the surface of the wood specimens increased the fungi colonization of the specimens; however, immersion of the specimens into the solution of corn steep liquor did not have the same positive influence. These results are important in elucidating copper toxicity in wood decay fungi and for using these fungi for bioremediation of treated wood wastes.

Impact of microbial activity on copper, lead and nickel mobilization during the bioremediation of soil PAHs

A fungal bioremediation method using P. frequentans removed up to 75% of phenanthrene with the addition of water and nutrients over a period of 30 d. During the bioremediation process, changes in metal behavior were monitored by an in situ technique (diffusive gradients in thin-films, DGT) and by soil solution chemistry. DGT provided absolute data on fluxes from the solid phase to the DGT device and relative trends of concentrations of the most labile metal species. DGT response indicated that bioremediation increases metal mobilization from the solid phase. Filtration provided data on the concentrations of solution phase (<0.45 μm) metal. In all case, metal fluxes and concentrations significantly increased after the bioremediation process began. Fluxes increased from <0.1 pg cm −2 s −1 before bioremediation to between 0.2 and 0.5 pg cm −2 s −1 after bioremediation. Metal concentrations in the soils solution (filtration at 0.45 μm) increased from 2 to 10 μg l −1 (Cu), 1–4 μg l −1 (Pb) and from 40 to 140 μg l −1 (Ni) after bioremediation. Although over a short time period, these data strongly indicated that there was remobilization of metal from solid to solution (and thus to the DGT device) directly due to the bioremediation process. Although the mechanism was not unambiguously identified, it was shown not to be related to small changes in bulk pH over time and was attributed to the microbial action on the surface of the soil solid phase, releasing metal into solution. Additionally, differences in metal concentration and flux were observed in sterilized and non-sterilized soils and in the absence or presence of phenanthrene. The results indicated that the bioremediation of soil by P. frequentans increased the flux, lability and mobility of trace metal species and therefore the likely metal bioavailability to plants.

Fungal Diversity and Use in Decomposition of Environmental Pollutants

This article presents a critical review of the actual state of fungal activities on environmental pollutants, fungal diversity, the use of fungi in the degradation of chemical pollutants, enzyme degrading systems and perspectives on the use of fungi in bioremediation and unexplored research. The ability of fungi to transform or metabolize chemical pollutants has received much attention due to environmental persistence and chemical toxicity. The fungal degradation of xenobiotics is looked upon as an effective method of removing these pollutants from the environment by a process which is currently known as bioremediation. This review summarizes information from fundamental works that have revealed that a wide variety of fungi are capable of degrading an equally wide range of toxical chemical. The capacity of non-ligninolytic and ligninolytic fungi in the bioremediation of polycyclic aromatic hydrocarbon (PAHs), benzene-toluene-ethylbenzene-xylene (BTEX), chlorophenols, polychlorinated biphenyl, munitions waste and pesticides have been discussed. Besides this, several extracellular enzymes are involved in the metabolism of xenobiotic compounds as well as other factors related to these processes.

Ligninolytic fungi in bioremediation: extracellular enzyme production and degradation rate

Ligninolytic fungi can be used for remediation of pollutants in water and soil. Extracellular peroxidases and laccases have been shown to oxidize recalcitrant compounds in vitro but the likely significance of individual enzyme levels in vivo remains unclear. This study documents the amounts and activities of Mn-dependent peroxidase (MnP), lignin peroxidase and laccase (LAC) in various species of ligninolytic fungi grown in liquid medium and soil and their effect on degradation of polycyclic aromatic hydrocarbons (anthracene and pyrene), a polychlorinated biphenyl mixture (Delor 106) and a number of synthetic dyes. Stationary cultures of a highly degradative strain Irpex lacteus exhibited 380-fold and 2-fold increase in production of MnP and LAC, respectively, compared to submerged cultures. Addition of Tween 80 to the submerged culture increased MnP levels 260-fold. High levels of MnP correlated with efficient decolorization of Reactive Orange 16 azo dye but not of Remazol Brilliant Blue R anthraquinone dye. Degradation of anthracene and pyrene in spiked soil by straw-grown explorative mycelium of Phanerochaete chrysosporium, Trametes versicolor and Pleurotus ostreatus showed the importance of MnP and LAC levels secreted into the soil. The importance of high fungal enzyme levels for efficient degradation of recalcitrant compounds was better demonstrated in liquid media compared to the same strains growing in soil.