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To evaluate the probiotic characteristics and safety of  Enterococcus durans  isolate A8-1 from a fecal sample of a healthy Chinese infant, we determined the tolerance to low pH, survival in bile salts and NaCl, adhesion ability, biofilm formation, antimicrobial activity, toxin gene distribution, hemolysis, gelatinase activity, antibiotic resistance, and virulence to  Galleria mellonella  and interpreted the characters by genome resequencing. Phenotypically,  E. durans  A8-1 survived at pH 5.0 in 7.0% NaCl and 3% bile salt under aerobic and anaerobic condition. The bacterium had higher adhesion ability toward mucin, collagen, and Bovine Serum Albumin (BSA)  in vitro  and showed high hydrophobicity (79.2% in chloroform, 49.2% in xylene), auto-aggregation activity (51.7%), and could co-aggregate (66.2%) with  Salmonella typhimurium . It had adhesion capability to intestinal epithelial Caco-2 cells (38.74%) with moderate biofilm production and antimicrobial activity against several Gram-positive pathogenic bacteria. A8-1 can antagonize the adhesion of  S. typhimurium  ATCC14028 on Caco-2 cells to protect the integrity of the cell membrane by detection of lactate dehydrogenase (LDH) and AKP activities. A8-1 also helps the cell relieve the inflammation induced by lipopolysaccharide by reducing the expression of cytokine IL-8 ( P  = 0.002) and TNF-α ( P  &amp;gt; 0.05), and increasing the IL-10 ( P  &amp;lt; 0.001). For the safety evaluation, A8-1 showed no hemolytic activity, no gelatinase activity, and had only  asa1  positive in the seven detected virulence genes in polymerase chain reaction (PCR), whereas it was not predicted in the genome sequence. It was susceptible to benzylpenicillin, ampicillin, ciprofloxacin, levofloxacin, moxifloxacin, tigecycline, nitrofurantoin, linezolid, vancomycin, erythromycin, and quinupristin/dalofopine except clindamycin, which was verified by the predicted  lasA ,  lmrB ,  lmrC , and  lmrD  genes contributing to the clindamycin resistance. The virulence test of  G. mellonella  showed that it had toxicity lower than 10% at 1 × 10 7  CFU. According to the results of these evaluated attributes,  E. durans  strain A8-1 could be a promising probiotic candidate for applications.

Probiotic Potential Analysis and Safety Evaluation of Enterococcus durans A8-1 Isolated From a Healthy Chinese Infant