An original liquid medium and a field broth bottle method for the rapid detection of the most probable number of sulfide-producing bacteria (SPB) from sulfate and thiosulfate are described. The broth bottle method enables after inoculation with a sample (water, sediment) the growth of ubiquitous aerobic bacteria, causing oxygen depletion, required for the growth of the present various anaerobic and facultative anaerobic SPB. The medium regularly gives higher numbers of the SPB than the used control medium (Postgate’s E) for detection of sulfate-reducing bacteria and the final results are obtained just 36 h after the medium inoculation. The method is simple and suitable for the estimation of the physiological group of SPB in fresh waters, saline waters, sediments and industrial waters.