Oil reservoir souring by the microbial reduction of sulfate to sulfide is unwanted, because it enhances corrosion of metal infrastructure used for oil production and processing. Reservoir souring can be prevented or remediated by the injection of nitrate or biocides, although injection of biocides into reservoirs is not commonly done. Whether combined application of these agents may give synergistic reservoir souring control is unknown. In order to address this we have used up-flow sand-packed bioreactors injected with 2 mM sulfate and volatile fatty acids (VFA, 3 mM each of acetate, propionate and butyrate) at a flow rate of 3 or 6 pore volumes (PV) per day. Pulsed injection of the biocides glutaraldehyde (Glut), benzalkonium chloride (BAC) and cocodiamine was used to control souring. Souring control was determined as the recovery time (RT) needed to re-establish an aqueous sulfide concentration of 0.8–1 mM (of the 1.7–2 mM before the pulse). Pulses were either for a long time (120 h) at low concentration (long-low) or for a short time (1 h) at high concentration (short-high). The short-high strategy gave better souring control with Glut, whereas the long-low strategy was better with cocodiamine. Continuous injection of 2 mM nitrate alone was not effective, because 3 mM VFA can fully reduce both 2 mM nitrate to nitrite and N2 and, subsequently, 2 mM sulfate to sulfide. No synergy was observed for short-high pulsed biocides and continuously injected nitrate. However, use of continuous nitrate and long-low pulsed biocide gave synergistic souring control with BAC and Glut, as indicated by increased RTs in the presence, as compared to the absence of nitrate. Increased production of nitrite, which increases the effectiveness of souring control by biocides, is the most likely cause for this synergy.