The microorganisms thriving in the ruminant’s digestive tract deliver the metabolic potential for degrading recalcitrant lignocellulose-rich organic matter and rendering it digestible. Amongst the microbiota thriving in the rumen, anaerobic fungi (AF) with the unpronounceable name Neocallimastigomycota host a complex array of enzymes that are especially adapted for degradation of lignocellulosic biomass like straw. In addition, their appressoria attach to the macromolecule structures and physically penetrate them, thus enhancing the enzymatic action.Cultivation of these fungi has often ended in frustration for researchers as the understanding of cultivation requirements is very poor. However, scientists see the great potential of these organisms for biotechnological use, in particular for the utilization of lignocellulosic residues (LCR) in closed loop recycling management. For this project, a consortium from Austria (leading agency; PI: Dr. Sabine Marie Podmirseg), Germany (PI: Dr. Michael Lebuhn) and Switzerland (PI: Prof. Urs Baier), all engaged in the harnessing of AF, is cooperating in close connection with research groups in the United Kingdom and in the Czech Republic. The overarching aim of the proposed project will be to remove existing methodological obstacles preventing the biotechnological utilization of anaerobic fungi. For that purpose, the consortium starts with the crucial basics, explores the cultivation requirements of Neocallimastigomycota, develops suitable culturing and detection methods and finally evaluates their biotechnological application for LCR disintegration.The working hypotheses are, amongst others: (i) the ecological niche for AF is larger than assumed to date; (ii) distinct requirements for micronutrients or supplements have to be met for long-time cultivation; (iii) cultivation without rumen fluid is possible for many strains; (iv) novel AF can be isolated from various also non-rumen habitats. The project will combine classical microbiological methods (microscopy, batch and continuous cultivation, enzymology) with molecular approaches (fluorescence in situ hybridization, quantitative PCR, direct RNA and next generation sequencing). It is expected to develop suitable protocols for the cultivation of anaerobic fungi and provide the basics for up-scaled production. The project’s goal is to enable researchers and the society to make use of the AF for energetic and material use of LCR.