[Objective] We studied the influences of water pressure and temperature on denitrification, and detected its nitrogen removal characteristics for providing evidence to remediate the micro-polluted reservoir source water. [Methods] Mixed oligotrophic aerobic denitrification bacteria was obtained through enrichment, domestication, and screening processes, which was isolated from sediment in the source water reservoir; and the nitrogen removal characteristics was detected by an in-situ biological inoculation experiment (DO at 3-8 mg/L). [Results] Nitrate of the hard flask system (with water pressure influence) was removed completely, however, at 0.5, 5 m water layer, the nitrate removal rate of the soft flask reached 90.66%, 100%, other layers reached 99.61%, 80.55%, 67.01%, 64.73%. No nitrite accumulated. Because of bacteria death, ammonia had a slight increase. At the end of the experiment, in the 0.5, 5.0, 7.5, 10.0, 12.5 and 15.0 m water layer, the total nitrogen removal rates of hard flask reached 50.11%, 61.49%, 56.24%, 44.50%, 36.80% and 38.73%, however, that of soft system reached 33.47%, 60.61%, 43.98%, 36.28%, 27.52% and 28.57%. OD₆₀₀ and pH first rose and then dropped. The mixed bacteria had prominent nitrogen removal ability between 11℃ and 30℃. [Conclusion] The mixed bacteria have a strong adaptability to temperature and the water pressure has a disadvantage to the nitrogen removal.