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Abstract Background Rabies (RABV) is an enzootic disease in Tunisia, with dogs being the primary reservoir. Vaccinating dogs is the key to eradicate rabies. Regional Veterinary Services conduct nationwide immunisation campaigns on an annual basis. Evaluation of the immune response is still important to make sure that the vaccination is effective in the conditions of the Tunisian field. In this paper, the FAVN technique was used to test rabies antibody dynamics in dogs from three distinct Tunisian areas observed for one year following a mass vaccination campaign. Results On day 30 after vaccination, 75% of all dogs vaccinated during the campaign were sero-positive (titres greater than or equal to 0.5 transformed IU/ml). On day 180, 48% of all dogs were sero-positive. Only 25.6% of primary-vaccinated dogs remained sero-positive on day 180 and 7% on day 365, whereas 91% of previously sero-positive dogs remained sero-positive on day 365. Conclusions Although a single rabies vaccine is successful at stimulating an immunological response, it is recommended that primary-vaccinated dogs have a second booster between one and three months after the initial vaccination to maintain seropositivity. To achieve the rabies eradication objective, all dogs should receive an annual booster to maintain effective immunological protection.