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In recent years, flathead grey mullets (<i>Mugil cephalus</i>) cultured in Eilat (Israel) have been highly affected by <i>Vibrio harveyi</i>, showing neurological signs such as uncoordinated circular swimming followed by high mortality rates. Despite the advances in and different approaches to control vibriosis associated with <i>Vibrio harveyi</i>, including commercial vaccines, most of them have not succeeded in long-term protection. In this study, we evaluated the effectiveness, long-term protection, and antibody production of three vaccine preparations: heat-killed bacteria (HKB), membrane proteins denaturation (BME PROT), and internal proteins (INT PROT) developed specifically against <i>Vibrio harveyi</i> for grey mullets. Our results show that fish immunized with heat-killed bacteria emulsified with adjuvant presented the most effective and long-lasting protection against the bacterium, and a cross-protection against other bacteria from the <i>harveyi</i> clade. The effectiveness of each immunization treatment correlated with the levels of specific antibody production against <i>Vibrio harveyi</i> in the serum of the immunized fish.