Development of cold-adapted microbial agent is an efficient approach for composting in low temperature. The study was conducted to evaluate the effect of semi-continuous replacements of compost materials after inoculation (SRMI) on the heat preservation of low temperature composting derived from chicken manure. Results revealed that SRMI could significantly improve the heat preservation of the pile, although the time of start-up in two inoculation groups was approximately the same. Due to the increase in the number of replacements of materials led to the changes in microbial community structures and enzyme activity. Non-metric multidimensional and colorimetric methods indicated that microbial community structures and enzyme activity was completely different in SRMI. Structural equation model was constructed by key factors involved in diversity of the microbial community, enzyme activity, temperature and bio-heat generation. In summary, SRMI decidedly increase the heat preservation time of the pile and start-up efficiency of the low temperature composting.