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The BOD (biochemical oxygen demand) sensor was fabricated by covering a dissolved oxygen (DO)probe with a microbe-impregnated membrane and a dialysis membrane. Various microorganisms isolated fromthe soils, water and activated sludge have been evaluated for measuring biochemical oxygen demand (BOD);Bacillus species HN24 and HN93 were selected as they exhibited rapid oxygen consumption and fast recovery.Improved BOD sensor could be prepared by using mixed microbes (Bacillus subtilis, Bacillus sp. HN24 andBacillus sp. NH93) and silicon rubber gas-permeable membrane for DO probe, and by bubbling 50% O2 (N2valence) through background bufer solution. This system exhibited excellent analytical performance resultingin good linearity (r2=0.9986) from 0 to 100 mg/L level of BOD.