Dissolved organic matter (DOM) derived from biochar takes a crucial role in transport and bioavailability toward contaminants; hence, it is undeniable that a thorough analysis of its properties is important. So far, the effect of pyrolysis temperature on the functional groups, components, and evolutionary sequence of manure-based biochar DOM has not been adequately investigated. Here, DOM was released from two typical livestock manures (cow and pig) at five pyrolysis temperatures (300 similar to 700 degrees C), and it was explored in depth with the aid of moving window 2D correlation spectroscopy (MW-2D-COS) and heterogeneous 2D correlation spectroscopy (hetero-2D-COS). The results demonstrated that the concentration, aromaticity, and hydrophobicity of DOM were greater at high temperatures, and more DOM was liberated from cow manure-based biochar at identical temperature. Protein-like compounds dominated at high temperatures. The pyrolysis temperatures of final configuration transformation points of the fulvic acid-like component and the aromatic ring C=C in DOM were 400 degrees C and 500 degrees C, respectively. Moreover, Fourier transform infrared spectroscopy combined with two-dimensional correlation analysis indicated that the functional group evolution of DOM depends on the pyrolysis temperature and feedstock type. The study provides a new perspective on manure management and environmental applications of biochar.