Probiotic Bacterium and Microalga Interaction on Rearing Kumamoto Oyster Crassostrea sikamea Spat.

Abstract

This study assessed in vitro interaction between Bacillus bacteria and microalgae and their posterior in vivo effect on rearing Kumamoto oyster Crassostrea sikamea. The probiotic strains Bacillus licheniformis (MAt32), B. subtilis (MAt43) and B. subtilis (GAtB1) were individually inoculated in triplicate into 250mL flasks containing 1 × 104 colony forming units (CFU) mL-1 of bacteria and 4.5 × 104 cell mL-1 of microalgae (Isochrysis galbana or Chaetoceros calcitrans) to evaluate their growth during a 7-day culture. Single cultures of microalgae or bacilli served as control. Additionally, C. sikamea spat was treated for 28days with four single/combined bacillus treatments in triplicate at a concentration of 1 × 106CFUmL-1 as follows: (a) control, without treatments; (b) combination of two antibiotics (10mg L-1); (c) B. licheniformis; (d) B. subtilis; (e) B. subtilis subtilis and (f) mixed bacilli. The results showed a significantly (P < 0.05) increased growth of Bacillus strains co-cultured with microalgae, while the growth of I. galbana co-cultured with bacteria was not reduced significantly (P > 0.05) compared with the control group. C. sikamea spat treated with Bacillus showed significantly (P < 0.05) higher growth and survival than the control group. In this study, C. calcitrans microalgae were susceptible to the presence of probiotic bacteria. Nonetheless, this reduction in microalgal growth observed in vitro increased growth and survival of C. sikamea spat exposed to probiotic bacteria when compared to spat without probiotics.