Nowadays, soil contamination by total petroleum hydrocarbons is still one of the most widespread forms of contamination. Intervention technologies are consolidated; however, full-scale interventions turn out to be not sustainable. Sustainability is essential not only in terms of costs, but also in terms of restoration of the soil resilience. Bioremediation has the possibility to fill the gap of sustainability with proper knowledge. Bioremediation should be optimized by the exploitation of the recent “omic” approaches to the study of hydrocarburoclastic microbiomes. To reach the goal, an extensive and deep knowledge in the study of bacterial and fungal degradative pathways, their interactions within microbiomes and of microbiomes with the soil matrix has to be gained. “Omic” approaches permits to study both the culturable and the unculturable soil microbial communities active in degradation processes, offering the instruments to identify the key organisms responsible for soil contaminant depletion and restoration of soil resilience. Tools for the investigation of both microbial communities, their degradation pathways and their interaction, will be discussed, describing the dedicated genomic and metagenomic approaches, as well as the interpretative tools of the deriving data, that are exploitable for both optimizing bio-based approaches for the treatment of total petroleum hydrocarbon contaminated soils and for the correct scaling up of the technologies at the industrial scale.

Recalcitrant compounds limit the efficiency of conventional biological processes for wastewater treatment, representing one of the major issues in the field. This study focused on the treatment of three effluents with White-Rot-Fungus (WRF) Bjerkandera adusta MUT 2295 in batch tests, with biomass cultivated in attached form on polyurethane foam cubes (PUFs) to test its efficiency in the removal of the target effluents’ recalcitrant fraction. Treatment efficiency of B. adusta was evaluated on landfill leachate (Canada) and two solutions containing synthetic recalcitrant compounds, which were prepared with tannic and humic acid. Chemical Oxygen Demand (COD) and color removal, the production of manganese peroxidases, and the consumption of a co-substrate (glucose) were monitored during the experiment. Biological Oxygen Demand (BOD₅) and fungal dry weight were measured at the beginning and at the end of the experiment. After co-substrate addition, effluent COD was 2300 ± 85, 2545 ± 84, and 2580 ± 95 (mg/L) in raw leachate and tannic and humic acids, respectively. COD removal of 48%, 61%, and 48% was obtained in raw leachate and in the synthetic effluents containing tannic and humic acids, respectively. Color removal of 49%, 25%, and 42% was detected in raw leachate and in tannic and humic acid solutions, respectively. COD and color removals were associated with the increase of fungal dry weight, which was observed in all the trials. These results encourage the use of the selected fungal strain to remove tannic acid, while further investigations are required to optimize leachate and humic acid bioremediation.

The aim of this study is to investigate the performance of a pilot-scale plug-flow reactor (PFR) as a biorefinery system to recover chemicals (i.e., volatile fatty acids (VFAs)), and biogas during the dry thermophilic anaerobic digestion (AD) of the organic fraction of municipal solid waste (OFMSW). The effects of the hydraulic retention time (HRT) on both outputs were studied, reducing the parameter from 22 to 16 days. In addition, VFA variation along the PFR was also evaluated to identify a section for a further valorization of VFA-rich digestate stream. A particular focus was dedicated for characterizing the community responsible for the production of VFAs during hydrolysis and acidogenesis. The VFA concentration reached 4421.8 mg/L in a section located before the end of the PFR when the HRT was set to 16 days. Meanwhile, biogas production achieved 145 NLbiogas/d, increasing 2.7 times when compared to the lowest HRT tested. Defluviitoga sp. was the most abundant bacterial genus, contributing to 72.7% of the overall bacterial population. The genus is responsible for the hydrolysis of complex polysaccharides at the inlet and outlet sections since a bimodal distribution of the genus was found. The central zone of the reactor was distinctly characterized by protein degradation, following the same trend of propionate production.

In the present study, an Ascomycete fungal strain, Lambertella sp., isolated from environmental polluted matrices, was tested for the capacity to reduce the contamination and the toxicity of intermediate and old landfill leachates. Batch tests in flasks, under co-metabolic conditions, were performed with two different old leachates, with suspended and immobilized Lambertella sp. biomass, resulting in a soluble chemical oxygen demand depletion of 70% and 45%, after 13 and 30 days, respectively. An intermediate landfill leachate was treated in lab-scale reactors operating in continuous conditions for three months, inoculated with immobilized Lambertella sp. biomass, in absence of co-substrates. The Lambertella sp. depleted the corresponding total organic carbon by 90.2%. The exploitability of the Lambertella sp. strain was evaluated also in terms of reduction of phyto-, cyto-, and mutagenicity of the different Landfill Leachates at the end of the myco-based treatment, resulting in an efficient depletion of leachate clastogenicity.

Innovative culturomic approaches were adopted to isolate hydrocarbonoclastic bacteria capable of degrading diesel oil, bitumen and a selection of polycyclic aromatic hydrocarbons (PAH), e.g., pyrene, anthracene, and dibenzothiophene, from a soil historically contaminated by total petroleum hydrocarbons (TPH) (10,347 ± 98 mg TPH/kg). The culturomic approach focussed on the isolation of saprophytic microorganisms and specialist bacteria utilising the contaminants as sole carbon sources. Bacterial isolates belonging to Pseudomonas, Arthrobacter, Achromobacter, Bacillus, Lysinibacillus, Microbacterium sps. were isolated for their capacity to utilise diesel oil, bitumen, pyrene, anthracene, dibenzothiphene, and their mixture as sole carbon sources. Pseudomonas, Arthrobacter, Achromobacter and Microbacterium sps. showed plant growth promoting activity, producing indole-3-acetic acid and expressing 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. In parallel to the culturomic approach, in the microbial community of interest, bacterial community metabarcoding and predictive functional metagenomic analysis were adopted to confirm the potentiality of the isolates in terms of their functional representativeness. The combination of isolation and molecular approaches for the characterisation of a TPH contaminated soil microbial community is proposed as an instrument for the construction of an artificial hydrocarbonoclastic microbiota for environmental restoration.

Dark fermentation (DF) is a simple method for hydrogen (H₂) production through the valorization of various organic wastes that can be used as feedstock. In particular, an organic fraction of municipal solid waste (OFMSW) is a fermentation substrate that can easily be gathered and provides high yields in biogas and value-added organic compounds such as volatile fatty acids (VFAs). DF is coupled with a methanogenic reactor to enhance biogas production from the OFMSW. In this study, a two-stage reactor was conducted and monitored to optimize the methane yield by reducing the HRT at the DF reactor. A focus of the functional inference based on a next-generation sequence (NGS) metabarcoding analysis and comparison of microbial communities that populate each reactor stage was performed. Concerning gas quality, the two-stage system observed a hydrogen-rich biogas in the first fermentative reactor (on average 20.2%) and an improvement in the methane content in the second methanogenic digester, which shifted from 61.2% obtained for the one-stage experiment to 73.5%. Such increases were due to the improvement in substrate hydrolysis. As for the specific biogas production, the results showed an overall increase of 50%.