Wood extractives promote pitch formation during pulp and paper manufacturing. To date, this problem has been controlled by extended storage of the chips and/or chemical additives. Biotreatment of the wood prior to pulping provides an alternative that not only decreases the negative impact of the extractives but may also improve the kraft pulping efficiency. This initiative seeks to verify the quantity and chemical composition of Eucalyptus nitens wood extractives following biotreatment with three albino fungi species (Ophiostoma floccosum, Ophiostoma piceae and Ophiostoma piliferum). Eucalyptus nitens wood chips were sprayed with spore suspensions of Ophiostoma piliferum, Ophiostoma piceae and Ophiostoma floccosum albino strains (1 × 108 spore concentration). After 7 and 21 days of fungal treatment, the extractive content was determined via Soxhlet extraction with an 80:20% n-hexane:ethyl acetate solvent mixture. The Ophiostoma floccosum F1A94, Ophiostoma piliferum F2D8 and Ophiostoma piceae F2A68 strains proved to be most capable of bioreduction with reductions of 35.1%, 33.2% and 29.3%, respectively. The chemical composition of the extract was analyzed via gas chromatography coupled with mass spectrometry, which demonstrated that most of the tested strains could reduce the β-sitosterol content.