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Summary  The lactoferricin expressed in  Bacillus subtilis  is relatively low in yield, making it hard to apply in industrial settings. We constructed a six tandem repeat of lactoferricin cDNA driven by promoter PtrnQ. After transformation, two transformants P245 and P263 possessing a stable inheritance of plasmid and high expression of lactoferricin were selected. The bactericidal activities, 1 μl of aliquot of a total 5.5 ml of solution extracted from 5 ml of cultured P245 and P263, were equivalent to the efficacy of 238.25 and 322.7 ng of Ampicillin against  Escherichia coli , respectively, and 366.4 and 452.52 ng of Ampicillin against  Staphylococcus epidermidis  respectively. These extracts were able to kill an Ampicillin‐resistant  E. coli  strain. The bactericidal activities of P245 and P263 equivalent to the efficacy of Tetracycline against  Vibrio parahaemolyticus  and  V. alginolyticus  were also determined. Moreover, the bactericidal activities of P245 and P263 were 168.04 and 249.94 ng of Ampicillin against  Edwardsiella tarda , respectively, and 219.7 and 252.43 ng of Tetracycline against  Streptococcus iniae  respectively. Interestingly, the survival rate of  E. tarda ‐infected tilapia fry fed the P263 extract displayed a significantly greater than that of the fry‐fed control strain. Collectively, these  B .  subtilis  transgenic strains are highly promising for use in animal husbandry during a disease outbreak.

High antimicrobial activity of lactoferricin‐expressing Bacillus subtilis strains