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This study was conducted to suggest effective storage methods for the use spent mushroom (Flammulina velutipes) substrates (SMS) as animal feed. SMS was storage by deep stacking, composting and vinyl mulching after treated with 1% microbes and 0.5% formic acid. During the storage periods, all of the treatment was covered with hyphae of mushroom. The external change of SMS was shown that the storage of SMS can be improved by anaerobic condition and 1% microbial treatment. The comparatively high temperature, pH and total bacterial number was shown in the SMS treated without microbes and formic acid by composting.