The purpose of this study was to develop a pilot scale bio-filter system removing ammonia gas with microorganisms. The system consisted of chaff(filter medium), a blower, a temperature sensor, a moisture sensor, a solenoid valve, and a heating system. Temperature and moisture contents were controlled via a PC to provide the microorganisms with proper environment. The microorganisms used in this study were Bacillius. coagulans NLRI T-6 and Pseudononas. putida NLRI S-21 of bacilli. Performance tests were performed to evaluate gas removal rate during 20 days. The result was shown that the removal rate was high in early days and gradually dropped below 90% without injecting the microbes. However, it was shown that when injecting the microbes, the removal rate was almost 100% and pH value was maintained at between 7 and 9 during the whole twenty-day period.