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In this study, two bacterial species, Salipiger thiooxidans and Exiguobacterium aestuarii, were extracted and screened from the Saemangeum Reservoir. This study examined these species’ suitability as a probiotic by confirming the effects of S. thiooxidans and E. aestuarii added to rearing water for L. vannamei. Three experimental groups were evaluated for 6 weeks: (1) a control group reared in natural (i.e., untreated) water (CON), (2) an experimental group in which S. thiooxidans was added to natural water (SMG-A), and (3) natural water inoculated with E. aestuarii (SMG-B). The SMG-B group inoculated with E. aestuarii showed significantly higher final body weight, weight gain, specific growth rates, and feed efficiency than the control group. The SMG-B group inoculated with E. aestuarii exhibited significantly higher levels of serum lysozyme, and ACP and ALP activity than the control and SMG-A groups. The SMG-A and SMG-B groups inoculated with probiotics showed significantly lower total ammonia nitrogen and nitrite than the control group. Our findings suggest that S. thiooxidans and E. aestuarii extracted from the Saemangeum Reservoir can improve the water quality of aquaculture water, and, in particular, E. aestuarii is a potential probiotic for L. vannamei.