Andrea Ceci

Uranium phosphate biomineralization by fungi

Geoactive soil fungi were investigated for phosphatase-mediated uranium precipitation during growth on an organic phosphorus source. Aspergillus niger and Paecilomyces javanicus were grown on modified Czapek-Dox medium amended with glycerol 2-phosphate (G2P) as sole P source and uranium nitrate. Both organisms showed reduced growth on uranium-containing media but were able to extensively precipitate uranium and phosphorus-containing minerals on hyphal surfaces, and these were identified by X-ray powder diffraction as uranyl phosphate species, including potassium uranyl phosphate hydrate (KPUO6 .3H2 O), meta-ankoleite [(K1.7 Ba0.2 )(UO2 )2 (PO4 )2 .6H2 O], uranyl phosphate hydrate [(UO2 )3 (PO4 )2 .4H2 O], meta-ankoleite (K(UO2 )(PO4 ).3H2 O), uramphite (NH4 UO2 PO4 .3H2 O) and chernikovite [(H3 O)2 (UO2 )2 (PO4 )2 .6H2 O]. Some minerals with a morphology similar to bacterial hydrogen uranyl phosphate were detected on A. niger biomass. Geochemical modelling confirmed the complexity of uranium speciation, and the presence of meta-ankoleite, uramphite and uranyl phosphate hydrate between pH 3 and 8 closely matched the experimental data, with potassium as the dominant cation. We have therefore demonstrated that fungi can precipitate U-containing phosphate biominerals when grown with an organic source of P, with the hyphal matrix serving to localize the resultant uranium minerals. The findings throw further light on potential fungal roles in U and P biogeochemistry as well as the application of these mechanisms for element recovery or bioremediation.

Phosphatase‐mediated bioprecipitation of lead by soil fungi

Geoactive soil fungi were examined for their ability to release inorganic phosphate (Pi ) and mediate lead bioprecipitation during growth on organic phosphate substrates. Aspergillus niger and Paecilomyces javanicus grew in 5 mM Pb(NO3)2-containing media amended with glycerol 2-phosphate (G2P) or phytic acid (PyA) as sole P sources, and liberated Pi into the medium. This resulted in almost complete removal of Pb from solution and extensive precipitation of lead-containing minerals around the biomass, confirming the importance of the mycelium as a reactive network for biomineralization. The minerals were identified as pyromorphite (Pb5(PO4)3Cl), only produced by P. javanicus, and lead oxalate (PbC2O4), produced by A. niger and P. javanicus. Geochemical modelling of lead and lead mineral speciation as a function of pH and oxalate closely correlated with experimental conditions and data. Two main lead biomineralization mechanisms were therefore distinguished: pyromorphite formation depending on organic phosphate hydrolysis and lead oxalate formation depending on oxalate excretion. This also indicated species specificity in biomineralization depending on nutrition and physiology. Our findings provide further understanding of lead geomycology and organic phosphates as a biomineralization substrate, and are also relevant to metal immobilization biotechnologies for bioremediation, metal and P biorecovery, and utilization of waste organic phosphates.

Phenotype MicroArray™ system in the study of fungal functional diversity and catabolic versatility

Fungi cover a range of important ecological functions associated with nutrient and carbon cycling in leaf litter and soil. As a result, research on existing relationships between fungal functional diversity, decomposition rates and competition is of key interest. Indeed, availability of nutrients in soil is largely the consequence of organic matter degradation dynamics. The Biolog® Phenotype MicroArrays™ (PM) system allows for the testing of fungi against many different carbon sources at any one time. The use and potential of the PM system as a tool for studying niche overlap and catabolic versatility of saprotrophic fungi is discussed here, and examples of its application are provided.

Transformation of vanadinite [Pb5(VO4)3Cl] by fungi

Saprotrophic fungi were investigated for their bioweathering effects on the vanadium- and lead-containing insoluble apatite group mineral, vanadinite [Pb5 (VO4 )3 Cl]. Despite the insolubility of vanadinite, fungi exerted both biochemical and biophysical effects on the mineral including etching, penetration and formation of new biominerals. Lead oxalate was precipitated by Aspergillus niger during bioleaching of natural and synthetic vanadinite. Some calcium oxalate monohydrate (whewellite) was formed with natural vanadinite because of the presence of associated ankerite [Ca(Fe(2+) ,Mg)(CO3 )2 ]. Aspergillus niger also precipitated lead oxalate during growth in the presence of lead carbonate, vanadium(V) oxide and ammonium metavanadate, while abiotic tests confirmed the efficacy of oxalic acid in solubilizing vanadinite and precipitating lead as oxalate. Geochemical modelling confirmed the complexity of vanadium speciation, and the significant effect of oxalate. Oxalate-vanadium complexes markedly reduced the vanadinite stability field, with cationic lead(II) and lead oxalate also occurring. In all treatments and geochemical simulations, no other lead vanadate, or vanadium minerals were detected. This research highlights the importance of oxalate in vanadinite bioweathering and suggests a general fungal transformation of lead-containing apatite group minerals (e.g. vanadinite, pyromorphite, mimetite) by this mechanism. The findings are also relevant to remedial treatments for lead/vanadium contamination, and novel approaches for vanadium recovery.

Biotransformation of β-hexachlorocyclohexane by the saprotrophic soil fungus Penicillium griseofulvum.

β-Hexachlorocyclohexane (β-HCH) is a persistent organic pollutant (POP) of global concern with potentially toxic effects on humans and ecosystems. Fungal tolerance and biotransformation of toxic substances hold considerable promise in environmental remediation technologies as many fungi can tolerate extreme environmental conditions and possess efficient extracellular degradative enzymes with relatively non-specific activities. In this research, we have investigated the potential of a saprotrophic soil fungus, Penicillium griseofulvum Dierckx, isolated from soils with high concentrations of isomers of hexachlorocyclohexane, to biotransform β-HCH, the most recalcitrant isomer to microbial activity. The growth kinetics of the fungus were characterized after growth in stirred liquid Czapek-Dox medium. It was found that P. griseofulvum was able to grow in the presence of 1 mg L(-1) β-HCH and in stressful nutritional conditions at different concentrations of sucrose in the medium (0 and 5 g L(-1)). The effects of β-HCH and the toluene, used as a solvent for β-HCH addition, on P. griseofulvum were investigated by means of a Phenotype MicroArray™ technique, which suggested the activation of certain metabolic pathways as a response to oxidative stress due to the presence of the xenobiotics. Gas chromatographic analysis of β-HCH concentration confirmed biodegradation of the isomer with a minimum value of β-HCH residual concentration of 18.6%. The formation of benzoic acid derivatives as dead-end products of β-HCH biotransformation was observed and this could arise from a possible biodegradation pathway for β-HCH with important connections to fungal secondary metabolism.

Fungal Bioweathering of Mimetite and a General Geomycological Model for Lead Apatite Mineral Biotransformations

ABSTRACT Fungi play important roles in biogeochemical processes such as organic matter decomposition, bioweathering of minerals and rocks, and metal transformations and therefore influence elemental cycles for essential and potentially toxic elements, e.g., P, S, Pb, and As. Arsenic is a potentially toxic metalloid for most organisms and naturally occurs in trace quantities in soil, rocks, water, air, and living organisms. Among more than 300 arsenic minerals occurring in nature, mimetite [Pb5(AsO4)3Cl] is the most stable lead arsenate and holds considerable promise in metal stabilization for in situ and ex situ sequestration and remediation through precipitation, as do other insoluble lead apatites, such as pyromorphite [Pb5(PO4)3Cl] and vanadinite [Pb5(VO4)3Cl]. Despite the insolubility of mimetite, the organic acid-producing soil fungus Aspergillus niger was able to solubilize mimetite with simultaneous precipitation of lead oxalate as a new mycogenic biomineral. Since fungal biotransformation of both pyromorphite and vanadinite has been previously documented, a new biogeochemical model for the biogenic transformation of lead apatites (mimetite, pyromorphite, and vanadinite) by fungi is hypothesized in this study by application of geochemical modeling together with experimental data. The models closely agreed with experimental data and provided accurate simulation of As and Pb complexation and biomineral formation dependent on, e.g., pH, cation-anion composition, and concentration. A general pattern for fungal biotransformation of lead apatite minerals is proposed, proving new understanding of ecological implications of the biogeochemical cycling of component elements as well as industrial applications in metal stabilization, bioremediation, and biorecovery.

Saprotrophic soil fungi to improve phosphorus solubilisation and release: In vitro abilities of several species

Modern agriculture is dependent on phosphate rock (PR), which is a nonrenewable resource. Improvement of phosphorus (P) availability for crops in agricultural soils represents a key strategy to slow down the depletion of PR. The aim of this study was to identify potential P biofertilisers among saprotrophic fungal species. We tested 30 fungal strains belonging to 28 taxa (4 Zygomycota and 24 Ascomycota) and with different life strategies. The study showed that many saprotrophic fungi have the ability to mobilise P from insoluble forms according to a variety of mechanisms. Our results expand the pool of P solubilising fungal species, also suggesting a new solubilisation index and shedding light on parameters that could be basic in the selection of efficient soil P-biofertilisers fungi. Rhizopus stolonifer var. stolonifer, Aspergillus niger and Alternaria alternata were found to be the best performing strains in terms of amounts of TCP solubilisation.

Metabolic synergies in the biotransformation of organic and metallic toxic compounds by a saprotrophic soil fungus

The saprotrophic fungus Penicillium griseofulvum was chosen as model organism to study responses to a mixture of hexachlorocyclohexane (HCH) isomers (α-HCH, β-HCH, γ-HCH, δ-HCH) and potentially toxic metals (vanadium, lead) in solid and liquid media. The P. griseofulvum FBL 500 strain was isolated from polluted soil containing high concentrations of HCH isomers and potentially toxic elements (Pb, V). Experiments were performed in order to analyse the tolerance/resistance of this fungus to xenobiotics and to shed further light on fungal potential in inorganic and organic biotransformations. The aim was to examine the ecological and bioremedial potential of this fungus verifying the presence of mechanisms that allow it to transform HCH isomers and metals under different extreme test conditions. To our knowledge, this work is the first to provide evidence on the biotransformation of HCH mixtures, in combination with toxic metals, by a saprotrophic non-white-rot fungus and on the metabolic synergies involved.

The Geological Roles Played by Microfungi in Interaction with Sulfide Minerals from Libiola Mine, Liguria, Italy

ABSTRACT The potential role played by fungi in the weathering of sulfide abandoned mines and waste rock dumps is scarcely investigated, yet. In particular microfungi may produce biofilms that work as sites of metals and minerals precipitation. This study aimed to investigate interactions, bioalteration, and biocorrosion between three microfungi (Trichoderma harzianum group, Penicillium glandicola, P. brevicompactum) isolated from the Libiola sulfide mine (Liguria, Italy) and pyrite-rich mineralizations occurring within the waste rock dumps. After six weeks of incubation, Environmental Scanning Electron Microscope (ESEM) analyses showed how single pyrite crystals were completely corroded and altered by all the selected species. These results represent the first step to establish that fungi play a central role in the biogeochemical cycles of extreme and contaminated sites such as sulfide mines, and that they actively contribute to the evolution of the degraded ecosystem to more harmonized scenery.

Roles of saprotrophic fungi in biodegradation or transformation of organic and inorganic pollutants in co-contaminated sites

For decades, human activities, industrialization, and agriculture have contaminated soils and water with several compounds, including potentially toxic metals and organic persistent xenobiotics. The co-occurrence of those toxicants poses challenging environmental problems, as complicated chemical interactions and synergies can arise and lead to severe and toxic effects on organisms. The use of fungi, alone or with bacteria, for bioremediation purposes is a growing biotechnology with high potential in terms of cost-effectiveness, an environmental-friendly perspective and feasibility, and often representing a sustainable nature-based solution. This paper reviews different ecological, metabolic, and physiological aspects involved in fungal bioremediation of co-contaminated soils and water systems, not only addressing best methods and approaches to assess the simultaneous presence of metals and organic toxic compounds and their consequences on provided ecosystem services but also the interactions between fungi and bacteria, in order to suggest further study directions in this field.