Alfalfa (Medicago sativa L.) production in the Coastal Plain of the southeastern United States requires enhanced soil fertility management to capitalize on increased dry matter yield and nutritive value. Poultry litter (PL) is an abundant fertilizer source that has been recognized as an economical choice for pasture and hay production. A field trial was established in east‐central Mississippi in which PL and inorganic fertilizer sources were compared in three alfalfa cultivars in 2018 and 2019. Fertilizer treatments included (a) 2 ton PL acre−1 (Treatment 1); (b) 4 ton PL acre−1 (Treatment 2); (c) inorganic N, P, and K delivered at similar rates to Treatment 1; and (d) inorganic P, and K (no N) supplied at same rates as Treatment 3. The cultivars were Bulldog 505, Bulldog 805, and AlfaGraze 600RR. Forage mass (FM), plant persistence (PER), nutritive value [crude protein (CP), total digestible nutrients (TDN), and relative forage quality (RFQ)], and economic comparisons were conducted. Repeated measures analysis observed no differences between cultivars or treatments for FM, PER, or CP. Bulldog 505 had superior TDN and RFQ values for 7 out of 11 total harvests. Plots with applied PL (2 or 4 ton acre−1) recorded greater TDN values than inorganic sources in 4 of 11 harvests. In 2018, Bulldog 505 produced the lowest cost per ton ($96.29 ton−1); in 2019, similar costs were observed between Bulldog 505 ($147.57 ton−1) and Bulldog 805 ($147.96 ton−1). Poultry litter sources resulted in the lowest cost per ton for both years of the trial.