Probiotics are widely applied in aquaculture, which can improve the growth, survival and health of aquatic animals. The aim of this study was to isolate and evaluate a Lactobacillus strain from the intestine of healthy whiteleg shrimp, Litopenaeus vannamei. A total of 20 Lactobacillus isolates showed rod-shaped morphology, gram-positive, non-motile, non-spore forming, and catalase and oxidase negative. Among the isolates, strain CMT1 exhibited the highest antibacterial activity against Vibrio parahaemolyticus and extracellular enzyme activity, which was selected and identified as Lactobacillus plantarum CMT1 based on 16S rDNA gene sequencing. This strain was also chosen to evaluate its effects on growth and survival of the whiteleg shrimp as a feed supplement. Four different diets were prepared, including the control (a commercial diet without L. plantarum CMT1) and three different concentrations of L. plantarum CMT1, viz. T1 (10⁶ CFU kg diet^-1), T2 (10⁷ CFU kg diet^-1), and T3 (10⁸ CFU kg diet^-1). After a 56-day feeding trial, the growth parameters in the T3 group had the highest values and showed a significant difference compared to the other groups. The survival rates of shrimp fed T2 and T3 diets were significantly higher than those in the control diet (p < 0.05). The lowest FCR value was recorded in the T3 group and showed a significant difference compared to the control group (p < 0.05). In addition, Lactobacillus counts in T2 and T3 groups increased significantly from day 28, which indicated that L. plantarum CMT1 could sustain its population in the intestine of shrimp. However, the total Vibrio sp. count in the T2 and T3 groups was significantly lower than that in the control group (p < 0.05). In conclusion, L. plantarum CMT1 could be used as a potential probiotic for shrimp aquaculture, and the recommended level of the probiotic strain CMT1 is 10⁸ CFU kg diet^-1.