Diversity, ecological role and potential biotechnological applications of marine fungi associated to the seagrass Posidonia oceanica.

The marine environment is characterized by high salinity and exerts a strong selective pressure on the biota, favouring the development of halo-tolerant microorganisms. Part of this microbial diversity is made up of fungi, important organisms from ecological and biotechnological points of view. In this study, for the first time, the qualitative and quantitative composition of the mycoflora associated to leaves, rhizomes, roots and matte of the seagrass Posidonia oceanica was estimated. A total of 88 fungal taxa, mainly belonging to Ascomycota, were identified by morphological and molecular methods. The most represented genera were Penicillium, Cladosporium and Acremonium. Most of the species (70) were selectively associated with one district; only two species (Penicillium chrysogenum var. chrysogenum and P. janczewskii) were isolated from all the districts. Moreover the capability to produce laccases, peroxidases and tannases by 107 fungal isolated by the different districts of P. oceanica was carried out. These results show that the mycoflora associated to P. oceanica is very rich and characterized by fungi able to produce ligninolytic enzymes and tannases useful to degrade and detoxify lignocellulose residues in presence of high salt concentrations. These fungi, hence, may play important ecological roles in marine environments but can also be very useful in different biotechnological areas.

Integrated fungal biomass and activated sludge treatment for textile wastewaters bioremediation.

A combined biological process was investigated for effective textile wastewater treatment. The process consisted of a first step performed by selected fungal biomasses, mainly devoted to the effluent decolourisation, and of a subsequent stage by means of activated sludge, in order to reduce the remaining COD and toxicity. In particular, the treatment with Trametes pubescens MUT 2400, selected over nine strains, achieved very good results in respect to all parameters. The final scale-up phase in a moving bed bioreactor with the supported biomass of the fungus allowed to verify the effectiveness of the treatment with high volumes. Despite promising results, further steps must be taken in order to optimize the use of these biomasses for a full exploitation of their oxidative potential in textile wastewater treatment.

Decolourisation and detoxification in the fungal treatment of textile wastewaters from dyeing processes.

In this study a selected fungal strain, Bjerkandera adusta (Willdenow) P. Karsten MUT 3060, was tested in different culture conditions to assess its real potential for bioremediation of textile wastewaters in terms of both decolourisation and detoxification. The fungus efficiently decolourised (colour removal up to 96%) four simulated wastewaters that mimic the recalcitrance of real ones for pH values, concentration of dyes, additives and salts. In the culture condition with the lowest N content, the decolourisation was coupled with an important detoxification of two simulated effluents, underlining the important influence of the cultural medium composition not only on the degradation but also on the detoxification of industrial wastes. In the other cases, despite an extensive decolourisation, no detoxification was observed. The fungus was further tested against a real effluent, collected from a wastewater treatment plant before and after the tertiary treatment (ozonation) to compare the two technologies in terms of chemical and toxicological parameters. The fungal treatment, although less efficient than ozonation, caused a good decolourisation of the effluent, with colour values within the threshold limits of the Italian law; both the fungal and the ozone treatment caused a detoxification, but only towards one of the three organisms used for the ecotoxicological tests. These results underline the crucial importance of the ecotoxicological analysis in assessing the applicability of a wastewater treatment.

Cunninghamella elegans biomass optimisation for textile wastewater biosorption treatment: an analytical and ecotoxicological approach.

The effect of pre-treatments on the composition of Cunninghamella elegans biomass and on its biosorption yields in the treatment of simulated textile wastewaters was investigated. The inactivated biomass was subjected to physical treatments, such as oven drying and lyophilisation, and chemical treatments using acid or alkali. The wastewater colour, COD and toxicity variations were evaluated. The lyophilisation sped up the biosorption process, whereas the chemical pre-treatment changed the affinity of biomass for different dyes. The alkali per-treated biomass achieved the highest COD reduction in the treatment of alkali wastewaters, probably because no release of alkali-soluble biomass components occurred under the alkaline pH conditions. Accordingly, only the acid pre-treated biomass decreased the COD of the acidic effluent. The ecotoxicity test showed significant toxicity reduction after biosorption treatments, indicating that decolourisation corresponds to an actual detoxification of the treated wastewaters. Fourier transform infrared spectroscopy, differential scanning calorimetry and thermogravimetric analyses of biomasses allowed highlighting their main chemical and physical properties and the changes induced by the different pre-treatments, as well as the effect of the chemical species adsorbed from wastewaters.

Survey of ectomycorrhizal, litter-degrading, and wood-degrading Basidiomycetes for dye decolorization and ligninolytic enzyme activity.

Basidiomycetes are essential in forest ecology, being deeply involved in wood and litter decomposition, humification, and mineralization of soil organic matter. The fungal oxidoreductases involved in these processes are today the focus of much attention with a view to their applications. The ecological role and potential biotechnological applications of 300 isolates of Basidiomycetes were assessed, taking into account the degradation of model dyes in different culture conditions and the production of oxidoreductase enzymes. The tested isolates belong to different ecophysiological groups (wood-degrading, litter-degrading, ectomycorrhizal, and coprophilous fungi) and represent a broad systematic and functional biodiversity among Basidiomycetes occurring in deciduous and evergreen forests of northwest Italy (Piedmont Region). The high number of species tested and the use of different culture conditions allowed the investigation of the degradation activity of several novel species, neglected to date. Oxidative enzyme activities varied widely among all ecophysiological groups and laccases were the most commonly detected enzymes. A large number of isolates (86%), belonging to all ecophysiological groups, were found to be active against at least one model dye; the wood-degrading fungi represented the most efficient group. Noteworthily, also some isolates of litter-degrading and ectomycorrhizal fungi achieved good decolorization yield. The 25 best isolates were then tested against nine industrial dyes commonly employed in textile industries. Three isolates of Bjerkandera adusta efficiently decolorized the dyes on all media and can be considered important candidates for application in textile wastewater treatment.

Industrial dye degradation and detoxification by basidiomycetes belonging to different eco-physiological groups.

Twenty-five basidiomycetes belonging to 17 species and ascribable to different eco-physiological groups were screened for their ability to decolorize 9 commercially important industrial dyes comprising a variety of anthraquinonic, azoic and phtalocyanin chromophores. The influence of the culture medium, particularly its C:N ratio, on decolourisation capacity was considered on solid substrate. Three strains of Bjerkandera adusta performed the highest decolourisation yields being able to degrade all dyes on all media and to produce a wide spectrum of oxidative enzyme activities. Hence, B. adusta strains were selected for further experiments in liquid cultures together with other 6 fungi that resulted effective in the decolourisation of the largest number of molecules in the broadest spectrum of cultural conditions. Particularly B. adusta MUT 3060 was found very effective (decolourisation percentage over 90%) in the treatment of simulated effluents composed of single and mixed dyes at high concentration (1000 mg/l). Peroxidase activity dependent (up to 362 U/l) and independent from manganese (up to 57 U/l) were detected during the decolourisation process. The Lemna minor toxicity test showed a significant reduction of toxicity after the fungal treatment indicating that decolourisation corresponded to an actual detoxification of the wastewater.

Scale-up of a bioprocess for textile wastewater treatment using Bjerkandera adusta.

Twelve basidiomycetes were investigated for their ability to degrade 13 industrial dyes and to treat four model wastewaters from textile and tannery industry, defined on the basis of discharged amounts, economic relevance and representativeness of chemical structures of the contained dyes. The best degradation yields were recorded for one strain of Bjerkandera adusta able to completely decolourise most of the dyes and to decolourise and detoxify three simulated wastewaters, showing a significant physiological versatility which is very useful for application purposes. The effects of different nutrient sources were investigated in order to optimize the yields of decolourisation and detoxification. Manganese-peroxidase and manganese-independent peroxidase were the only recorded enzymatic activities. In order to evaluate its true bioremediation potential, this strain was packed in a fixed-bed bioreactor, for treatment of large volumes of a real wastewater. The fungus resulted effective during 10 cycles of decolourisation, remaining active for a very long period, in non-sterile conditions.

Chromium removal from a real tanning effluent by autochthonous and allochthonous fungi.

Heavy metals represent an important ecological and health hazard due to their toxic effects and their accumulation throughout the food chain. Conventional techniques commonly applied to recover chromium from tanning wastewaters have several disadvantages whereas biosorption has good metal removal performance from large volume of effluents. To date most studies about chromium biosorption have been performed on simulated effluents bypassing the problems due to organic or inorganic ligands present in real industrial wastewaters that may sequestrate the Cr(III) ions. In the present study a tanning effluent was characterized from a mycological point of view and different fungal biomasses were tested for the removal of Cr(III) from the same tanning effluent in which, after the conventional treatments, Cr(III) amount was very low but not enough to guarantee the good quality of the receptor water river. The experiments gave rise to promising results with a percentage of removed Cr(III) up to 40%. Moreover, to elucidate the mechanisms involved in biosorption process, the same biomasses were tested for Cr(III) removal from synthetic aqueous solutions at different Cr(III) concentrations.

Pyrene degradation and detoxification in soil by a consortium of basidiomycetes isolated from compost: role of laccases and peroxidases.

A consortium of three basidiomycetes isolated from compost was investigated for pyrene degradation in soil microcosms. Pyrene concentration, glucose and ammonium evolution, moisture content, ligninolytic enzyme activities and phytotoxicity (germination index) on Lepidium sativum L. seeds were monitored. The fungal consortium grown on straw was found able to efficiently colonize soil and remove about 56 out of 100 mg kg(-1) of soil dry weight of pyrene in 28 days; in the meantime the germination index increased indicating a reduction of phytotoxicity. A glucose supply after 2 weeks was found useful to ensure fungal growth and activity; maintenance of moisture content below 70% allowed a good aeration of the system and improved degradation rates. Enzymatic assays showed that laccase and manganese independent peroxidase activity could have played a role in the degradation process.

Decolourisation and detoxification of textile effluents by fungal biosorption.

Textile effluents, in addition to high COD, display several problems mainly due to toxicity and recalcitrance of dyestuffs. Innovative technologies effective in removing dyes from large volumes of effluents at low cost and in a timely fashion are needed. Fungi are among the most promising organisms for dye biosorption. In this study dye decolourisation, COD and toxicity decrease of three wastewater models after the treatment with inactivated biomasses of three Mucorales fungi cultured on two different media were evaluated. Fungal biomasses displayed good sorption capabilities giving rise to decolourisation percentages up to 94% and decrease in COD up to 58%. The Lemna minor toxicity test showed a significant reduction of toxicity after biosorption treatments, indicating that decolourisation corresponds to an actual detoxification of the treated wastewaters.

Bioremediation potential of basidiomycetes isolated from compost.

The potential of a consortium of three basidiomycete mycelia isolated from compost to degrade polycyclic aromatic hydrocarbons (PAH) was first evaluated using a test based on decolorization of Poly R-478 dye. When pre-grown on straw, the consortium decolorized the dye by 83% in 7 days and generated a laccase activity of 663 IU l(-1). Its ability to degrade naphthalene was investigated in soil microcosms specially suited for this volatile PAH. The kinetic study was conducted at a maximal naphthalene concentration of 500 mg kg(-1) of soil. Naphthalene concentration, CO(2) evolution and phytotoxicity (germination index, GI%) on Lepidium sativum seeds were monitored. The naphthalene concentration decreased by about 70% in three weeks in the presence of metabolic activity, while the GI% increased indicating reduced phytotoxicity.

Isolation and identification of fungal communities in compost and vermicompost.

This research illustrates the qualitative and quantitative composition of the mycoflora of both a green compost (thermophilically produced from plant debris) and a vermicompost (mesophilically produced by the action of earthworms on plant and animal wastes after thermophilic preconditioning). Fungi were isolated using three media (PDA, CMC, PDA plus cycloheximide), incubated at three temperatures (24, 37 and 45 C). Substantial qualiquantitative differences in the species composition of the two composts were observed. The total fungal load was up to 8.2 X 10(5) CFU/g dwt in compost and 4.0 x 10(5) CFU/g dwt in vermicompost. A total of 194 entities were isolated: 118 from green compost, 142 from vermicompost; 66 were common to both. Structural characterization of this kind is necessary to determine the most appropriate application of a compost and its hygienic quality.