The pathogenicity and acute toxicity of Lactobacillus (L.) pentosus PL11 from eel (Anguilla japonica) were investigated using male and female Sprague-Dawley rats. The pathogenicity of L. pentosus PL11 was examined after treating the rats with 10∨11 CFU/mL, 10∨9 CFU/mL or 10∨7 CFU/mL doses of L. pentosus PL11 culture or 0.85% NaCl (Control) intragastrically. For acute toxicity studies, rats were treated with dried culture broth of L. pentosus PL11 at doses of 5,000 mg/mL, 2,500 mg/mL, 1,250 mg/mL or 625 mg/mL or Lactobacilli MRS broth (Control), and clinical signs or mortalities were monitored for two weeks. The results of the present investigation revealed no mortalities or obvious clinical signs in rats administered with the live bacterial cultures or dried culture broth at any investigated dose level. Also, no significant differences were observed in net body weight gain, gross pathological findings, feed and water consumption and body temperature among the different treatment groups and between the treated and control rats. It can be concluded from the above findings that L. pentosus PL11 is a safe probiotic strain with potential as feed additive to increase the feed efficiency or health of fish.