Improvement in the growth performance of white shrimp, Litopenaeus vannamei, by a protease-producing probiotic, Bacillus subtilis E20, from natto

Aims: To isolate and identify a benefic bacterium, Bacillus subtilis E20, from natto (fermented soybeans), and incorporate it into shrimp feed to promote shrimp growth performance. Methods and Results: A protease-producing bacterium, E20, isolated from natto was identified as B. subtilis by an API 50 CHB kit and the 16S rDNA sequence. B. subtilis E20 was able to grow at a broad range of temperatures (10-50 degrees C), pH values (5-10), and NaCl levels (0-9%). The best culture conditions for B. subtilis E20 to produce the protease were 40 degrees C, a pH of 6-8 and 0% NaCl. No shrimp died after being injected with B. subtilis E20 [up to 10(9) colony-forming units (CFU) per shrimp]. Bacillus subtilis E20 was incorporated in diets at the levels of 0 (control), 10(6), 10(7), and 10(8) CFU kg(-1) for shrimp grow-out culture, and results showed that after feeding on B. subtilis E20-containing diets (10(8) CFU kg(-1) of diet), shrimp had excellent growth performance and production compared to the control because protease activities in the digestive tract were improved by B. subtilis E20. Conclusions: Bacillus subtilis E20 isolated from natto is a great protease producer and is able to improve shrimp growth performance through increasing the digestibility of food. Significance and Impact of the Study: Results suggest that B. subtilis E20 is a potential candidate for use as a probiotic to improve shrimp growth performance, and consequently reduce feed costs.