The purpose of this research is to determine the bacteria that have the potential to degrade pertroleum in the waters of Ambon Bay. The research was conducted in July-September 2020 in the waters of Ambon Bay, precisely in two locations, namely PT. Perikanan Nusantara Ambon Branch Port in Galala (station 1) and Perikanan Nusantara Ambon Port in Tantui (station 2). The research method used was purposive sampling method on seawater sampling, laboratory-scale experimental method for the isolation of oil-decomposing bacteria, and morphological characterization observations using Stone Mineral Salt Solution Extract Yeast (SMSSe) media. The identification of bacteria uses macroscopic observation to observe the characteristics of the bacterial colony, while microscopic observation is used to observe the shape of the cell and the properties of the bacteria. Environmental parameters at the two stations include temperature, pH, DO, salinity, and brightness categorized as normal for bacterial growth. The results of bacterial isolation is that 7 isolates were obtained, consisting of 5 isolates found at station 1 and 2 isolates found at station 2. The highest total number of bacteria is 123.10-5 CFU/ml, while the lowest is 47.10-5 CFU/ml. In general, the characteristics of diesel oil-degrading bacterial isolates have circular and irregular colony forms, entire and lobate colony edges, raised and convex colony elevations, milky white, yellow, and cream colony colors, and bacilli and coccus cell shapes. Based on the results of gram isolate staining, there were 5gram negative bacterial isolates and 2gram positive bacterial isolates.

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