Bacillus 2종의 포스트바이오틱스 제작 조건 최적화 및 Edwardsiella piscicida와 Streptococcus parauberis에 대한 in vitro 항균 활성 평가
Optimization of postbiotic production conditions for two Bacillus strains and evaluation of in vitro antimicrobial activity against Edwardsiella piscicida and Streptococcus parauberis
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Probiotics have limitations such as the risk of inducing antibiotic resistance, release of live strains into aquatic environments, and difficulties in storage and transportation. To address these issues, this study aimed to optimize postbiotic production from shrimp powder derived Bacillus amyloliquefaciens FC820 (BA) and the reference strain Bacillus subtilis KCTC 1326 (BS). Both strains were cultured in MRS broth at 30℃ and 37℃ for 96 h and 120 h, after which the supernatants were harvested and heat-treated using a 100℃ water bath and a 105℃ dry oven. In water bath treatments, BA and BS were inactivated after 1 h and 45 min, respectively, whereas in dry oven treatments, inactivation occurred at 3 h for BA and 2.5 h for BS. When the supernatant cultured at 30℃ for 120 h was treated in a 100℃ water bath for 1 h, it produced the largest inhibition zones of 8.67 ± 0.29 mm and 12.17 ± 0.58 mm against Edwardsiella piscicida, and 16 mm and 15.17 ± 0.58 mm against Streptococcus parauberis, respectively. Cytotoxicity assays using EPC cells and CCK-8 showed that postbiotic concentrations up to 12.5% maintained cell viability above 90%, indicating safety at this dilution. This study underscores the potential of postbiotics as a sustainable and effective strategy for disease control and health management in aquaculture.
Abstract
Probiotics have limitations such as the risk of inducing antibiotic resistance, release of live strains into aquatic environments, and difficulties in storage and transportation. To address these issues, this study aimed to optimize postbiotic production from shrimp powder derived Bacillus amyloliquefaciens FC820 (BA) and the reference strain Bacillus subtilis KCTC 1326 (BS). Both strains were cultured in MRS broth at 30℃ and 37℃ for 96 h and 120 h, after which the supernatants were harvested and heat-treated using a 100℃ water bath and a 105℃ dry oven. In water bath treatments, BA and BS were inactivated after 1 h and 45 min, respectively, whereas in dry oven treatments, inactivation occurred at 3 h for BA and 2.5 h for BS. When the supernatant cultured at 30℃ for 120 h was treated in a 100℃ water bath for 1 h, it produced the largest inhibition zones of 8.67 ± 0.29 mm and 12.17 ± 0.58 mm against Edwardsiella piscicida, and 16 mm and 15.17 ± 0.58 mm against Streptococcus parauberis, respectively. Cytotoxicity assays using EPC cells and CCK-8 showed that postbiotic concentrations up to 12.5% maintained cell viability above 90%, indicating safety at this dilution. This study underscores the potential of postbiotics as a sustainable and effective strategy for disease control and health management in aquaculture.