This study was conducted to evaluate effects of mixed microbes addition on physico-chemical, fermentative and microbial parameters of sawdust-based spent mushroom substrates(SMS). The SMS was inoculated with mixed microbes(Enterobacter ludwigii, Bacillus cereus, 2 strains of Bacillus subtillis, Saccharomyces cerevisiae and Lactobacillus plantarum) at 1% level(wet basis) and anaerobically fermented during the different periods(up to 8th week). Compared with the SMS before ensiling, the ensiled one had higher CP, NDF and ADF percentages and lower DM and NFC percentages. However, levels of change were very low. The in situ ruminal disappearance of SMS DM and NDF decreased with the ensiling period prolonged. For fermentative parameters, pH reduced and lactic acid contents increased after ensiling, compared with those after ensiling. At 8th week of ensiling, pH increased and lactic acid contents reduced again, compared with those at 4th week of ensiling; however, the silage still showed favorable fermentation status. Lactic acid bacteria counts did not change throughout 8 weeks of ensiling. Counts of total microbes and yeast reduced after 4th week of ensiling period. Counts of lactic acid bacteria and yeast at 8th week of ensiling were in the levels of 108cfu/g. These results indicate that anaerobic fermentation with microbial addition could be an effective way for the long term(8 weeks) storage of the SMS.