The tendency of use of antibiotics is declining worldwide. From the perspect of disease prevention, the method is to use probiotics as a feed additive to induce the defense ability of fish and shrimp against the infection and disease. This study aims to evaluate the probiotic properties of four lactic acid bacteria (LAB) which were isolated from the intestinal gut of Seabastiscus marmoratus, Dentex tumifrons, Penaeus monodon, and Aristeus virilis. Those LAB were identified as Enterococcus sp. (F1), Lactobacillus sakei (F2), Lactobacillus farciminis (P1) and Leuconostoc sp. (P2). They all showed tolerance under the conditions of pH 3.0, 0.4% bile salt, and 3% NaCl, and could adhere to epithelioma papillosum cyprinid and Caco-2 cells. All strains demonstrated inhibitory effects against Vibrio pathogens (V. alginolyticus; V. parahaemolyticus; V. anguillarum), and were susceptible to chloramphenicol and tetracycline. Furthermore, the antioxidant activity of LAB showed effects on scavenging ability of ABTS radical, superoxide anion and chelating ability on ferrous ions.