The present study was conducted to identify sustainable alternative protein sources using locally available feedstuffs for Nile tilapia (Oreochromis niloticus L) production in three eco-regions of Kenya. Based on their availability, content of protein and fibre and the feasibility of removal of antinutritional factors, hydrolysed feather meal, boiled tea leaves residues, leaves of Ipomoea batatus, Manihot esculenta and Papaya carica were identified as most promising potential non-conventional feedstuffs. In a feeding experiment freshwater shrimp meal (Caridina nilotica; FSM) was gradually substituted by hydrolysed feather meal (HFM). Results indicated that substitution of FSM at HFM levels above 50% in aquaria led to significant growth reductions. However, complete substitution did not significantly affect growth of fish in low-input aquaculture. In conclusion, 100% substitution of FSM by HFM may be possible in semi-intensive culture of O. niloticus, where natural food is available. Both in pond and aquaria setup, the 50% diet gave the best performance overall and therefore has the highest potential for O.niloticus production. This is especially the case for a combination of the protein sources FSM, HFM and Pawpaw (Papaya carica) leaf meal, wich was tested in another growth experiment. From this study it can be concluded that locally available non-convectional feedstuffs can sustainably be utilized in O.niloticus production. Further research is to be directed towards analysing the effects of the feedstuffs identified herein on production traits of O.niloticus grown to market size.