Being a part of the large-scale Indus-Tsangpo suture zone ophiolites of the Alpine-Himalayan orogenic system, the Neo-Tethyan ophiolites of eastern Arunachal Himalaya, northeast India, are important in understanding the formation and evolution of the Neo-Tethyan lithosphere. This study focuses on the hornblendite dykes occurring within serpentinized peridotites of the Mayodia ophiolites in eastern Arunachal Himalaya, to ascertain the origin and evolution of such hydrous mineral cumulates within the ophiolite complex. Texturally and chemically, the hornblendites contain three types of amphiboles, Amp1 (pargasite), Amp2 (Mg-hornblende), and Amp3 (actinolite). The pargasites occur as large cm-sized phenocrysts and show cumulate textures while Mg-hornblende appears as both intercumulus grains between cumulus pargasites and surrounding partially resorbed clinopyroxene grains. Actinolite occurs along the breakdown zone of pargasite and is associated with other secondary minerals. The hornblendites are characterized by high MgO (~12–20 wt%) and low SiO2 (~42–44 wt%) with depleted light rare earth elements (REE) [(La/Sm)N = 0.29–0.52] and almost flat heavy REE [(Gd/Yb)N) = 0.89–1.11]. Moreover, the samples are characteristically depleted in high field strength elements (Nb, Ta, Ti, Hf) while being enriched in Pb. Geothermobarometric calculations yield high magmatic temperatures of ~950 and ~840 °C for pargasite and Mg-hornblende, respectively, and variable pressures with pargasite having the highest pressures (1.6–1.7 GPa) followed by Mg-hornblende (1.1–1.2 GPa) and finally actinolite (~0.55 GPa). Furthermore, fO2 calculations for the cumulus pargasites revealed highly oxidizing conditions (ΔNNO+2) and water content of parental melts (H2Omelt% = ~14.25 %). Parental melt calculations using whole-rock REE concentrations suggest formation from a hydrous sub-alkaline basaltic melt. Whole-rock initial SrNd isotopic ratios of the hornblendites reveal a primary signature for them, falling in the mantle array in proximity to the high-μ (HIMU) mantle reservoir. All these signatures point towards a magmatic arc affinity for these hornblendites. The host peridotite unit shows highly enriched REE concentrations (ƩREE = 18.13 ppm) with flat REE patterns compared to the other peridotites of the locality, which show depleted signatures. Olivine chemistry of the host peridotite unit depicts primary compositions falling in the mantle array. The olivine-liquid thermometer yields temperatures ~1000 °C, which are much lower than those of peridotites occurring in spreading centers (MOR or back-arc). These evidence suggest that the hornblendites and their host peridotite unit represent an island arc root complex, which further imply the hydrous nature of the Neo-Tethyan sub-arc mantle.