The effects of metal nanoparticle (NP) addition during anaerobic digestion (AD) of poultry litter was tested using two sequential experiments: Exp. A) four NPs (Fe, Ni, Co, and Fe3O4) at three concentrations; and Exp. B) NP combinations (Fe, Ni, and Co) at four concentrations. Scanning electronic microscopy (SEM) and elemental analysis were used to confirm NP inclusion after dispersion (before AD) and track nanoparticles post-AD, and new technique for NP extraction post-AD was developed. Before AD, NPs ranged from 30.0 to 80.9 nm for Fe, Ni, and Co, and 94.3 to 400 nm for Fe3O4. Methane production increased with NPs addition compared to poultry litter-only, with the highest increases observed with NPs concentrations (in mg/L) of 12 Ni (38.4% increase), 5.4 Co (29.7% increase), 100 Fe (29.1% increase), and 15 Fe3O4 (27.5% increase). Nanoparticle mixtures greatly decreased H2S production. The SEM post-AD detected Fe, Ni, and Fe3O4 at concentrations >= 100 mg/L.