Hydrocarbons can have very harmful effects on organisms and the environment, and conventional techniques for their removal are expensive and require the use of chemicals and long-term actions. Trichoderma is an ascomycete genus known to be active on different recalcitrant substrates, since it can produce a set of nonspecific extracellular enzymes generally involved in the degradation of lignin. However, the literature concerning the use of Trichoderma to degrade hydrocarbons is still limited. In this work we aimed to investigate the ability of Trichoderma to exploit used engine oil as its sole carbon source for prospective bioremediation of contaminated substrates. Four different strains belonging to Trichoderma asperellum and Trichoderma harzianum species were tested. The fungi were inoculated in direct contact with used engine oil, and after 45 days the samples were analyzed by gas chromatography/mass spectrometry (GC/MS). The results showed that all strains (except Trichoderma asperellum F1020) significantly changed the oil composition, decreasing the aromatic fraction in favor of the aliphatic one. T. harzianum F26, especially, showed a significant reduction of the BTEX (benzene, toluene, ethylbenzene, and the three xylene isomers) and alkylbenzenes fraction and an increase in short-chain aliphatics C₁–C₂₀. Enzymatic tests for laccase and peroxidase were also carried out, demonstrating that every strain seems to express a different mode of action.

Natural attenuation (NA) processes represent a valuable option in groundwater remediation. At a heavily 1,2-dichloroethane (1,2-DCA) contaminated site, Compound-Specific Isotope Analysis (CSIA) in combination with Biological Molecular Tools (BMTs) were implemented as a rigorous characterization approach to evaluate the occurrence of Natural Attenuation in the proximity of the source area. By the use of microcosm experiments, the potential for natural and enhanced biodegradation under anaerobic conditions was documented, following the dichloroelimination pathway. Enrichment factors of −9.1‰ and −11.3‰ were obtained for ¹³C while Geobacter spp. and reductive dehalogenase genes (rdhs) were identified as main site-specific biomarkers. At pilot scale, enrichments of 13.5‰ and 6.3‰ for δ¹³C and δ³⁷Cl, respectively, high levels of reductive dehalogenase (rdh group VI) along with the dominance of Geobacter spp. indicated the occurrence of significant dichloroelimination processes in groundwater under anaerobic conditions. By using the site-specific enrichment factors, degradation extents over approximately 70–80% were estimated, highlighting the relevant potential of NA in 1,2-DCA degradation in the vicinity of the source area at the site. The proposed fine-tuned protocol, including CSIA and BMTs, is proven to be effective as a groundwater remediation strategy, properly assessing and monitoring NA at site scale.