Even thoughdigestate, which is continually generated in anaerobic digestion process, can only be used as fertilizer during the growing season, digestate treatment is still a critical, environmental problem. That is why the present work aims to develop a method to manage digestate in agricultural biogas plant in periods when its use as fertilizer is not possible. A lab-scale system for the biological treatment of the digestate liquid fraction using the activated sludge method with a separate denitrification chamber was constructed and tested. The nitrogen load that was added tothe digestate liquid fraction accounted for 78.53% of the total nitrogen load fed into the reactor. External carbon sources, such as acetic acid, as well as flume water and molasses, i.e., wastewater and by-products from a sugar factory, were used to support the denitrification process. The best results were obtained using an acetic acid and COD (Chemical Oxygen Demand)/NO3–N (Nitrate Nitrogen) ratio of 7.5. The removal efficiency of TN (Total Nitrogen), NH4–N (Ammonia Nitrogen) and COD was 83.73%, 99.94%, 86.26%, respectively. It was interesting to see results obtained that were similar to those obtained when using flume water and COD/NO3–N at a ratio of 8.7. This indicates that flume water can be used as an alternative carbon source to intensify biological nitrogen removal from digestate.