The study focuses on managing microbial souring in alkali-surfactant-polymer (ASP) flooded reservoirs, a promising oil recovery technique. It identifies four alkaline-tolerant hydrogen sulfide-producing bacteria (SPB) strains with diverse sulfur metabolism. Nitrate partially inhibited some strains, while nitrite was more effective on others. Glutaraldehyde effectively prevented H2S production in both pure cultures and microcosmic simulation systems. This research aims to bridge the gap in souring management in high alkaline ASP-flooded reservoirs by proposing tailored strategies for preventing hydrogen sulfide production. The study also explores the sensitivity of SPB strains to nitrate, nitrite, and biocides, crucial for developing effective souring prevention technology in oil reservoirs. By comparing the diversity of SPB enriched in different culture media, the study sheds light on physiological characteristics and mitigation strategies for souring in ASP-flooded oil fields. Produced water samples from ASP-flooded reservoirs were collected, and noticeable differences were observed in the types of SPB enriched based on the nutrient components of the media used for enrichment.