Single cell proteins such as that of edible filamentous fungal biomass are considered as a promising sustainable source of animal feed supplementation. Filamentous fungi can be cultivated on different organic substrates including volatile fatty acids (VFAs) such as acetic, propionic, and butyric acids. These VFAs can be generated through the famous waste valorisation approach of anaerobic digestion (AD) as intermediate metabolites. This project investigates a sustainable approach for the production of animal feed supplementation through cultivation of fungal biomass on waste derived VFAs along with the in vitro analysis of fungal biomass digestibility as ruminant feed. In this regard, optimum conditions for the production of Aspergillus oryzae biomass on different VFAs effluents derived from anaerobic digestion process of food waste plus chicken manure (FWCKM) and potato protein liquor (PPL) at different pH, nitrogen sources, and feed mixture was studied. Accordingly, analyses showed that PPL has the highest biomass yield with 0.4 (g biomass/g consumed VFAs) based on the volatile solids (VS) by adjusting pH to 6.2. Furthermore, the digestibility of the produced fungal biomass is analysed by using three different in vitro digestion methods including Tilley and Terry (TT) method, Gas Production Method (GPM), and Nylon Bag Method (NBM) and the results are compared with the conventional feed (silage and rapeseed meal). Results obtained from different digestibility methods illustrate that different A. oryzae fungal biomass had approximately 10-15 % higher dry matter digestibility fraction compared to silage and rapeseed meal (reference feeds). Hence, these results revealed that A. oryzae fungal biomass can grow on VFAs effluents and produce protein-rich fungal biomass while this biomass has better digestibility compared to conventional feeds and confirmed the initial hypothesis of the study.