Safety Evaluation of Bifidobacterium breve IDCC4401 Isolated from Infant Feces for Use as a Commercial Probiotic

Abstract

Previously, our research group isolated Bifidobacterium breve IDCC4401 from infant feces as a potential probiotic. For this study, we evaluated the safety of B. breve IDCC4401 using genomic and phenotypic analyses. Whole genome sequencing was performed to identify genomic characteristics and investigate the potential presence of genes encoding virulence, antibiotic resistance, and mobile genetic elements. Phenotypic analyses including antibiotic susceptibility, enzyme activity, production of biogenic amines (BAs), and proportion of D-/L-lactate were evaluated using E-test, API ZYM test, high-performance liquid chromatography (HPLC), and D-/L-lactic acid assay respectively. The genome of B. breve IDCC4401 consists of 2,426,499 bp with a GC content of 58.70% and 2,016 coding regions. Confirmation of the genome as B. breve was provided by its 98.93% similarity with B. breve DSM20213. Furthermore, B. breve IDCC4401 genes encoding virulence and antibiotic resistance were not identified. Although B. breve IDCC4401 showed antibiotic resistance against vancomycin, we confirmed that this was an intrinsic feature since the antibiotic resistance gene was not present. B. breve IDCC4401 showed leucine arylamidase, cystine arylamidase, α-galactosidase, β-galactosidase, and α-glucosidase activities, whereas it did not show production of harmful enzymes such as β-glucosidase and β-glucuronidase. In addition, B. breve IDCC4401 did not produce any tyramine, histamine, putrescine, cadaverine, or 2-phenethylamine, which are frequently detected BAs during fermentation. B. breve IDCC4401 produced 95.08% of L-lactate and 4.92% of Dlactate. Therefore, our findings demonstrate the safety of B. breve IDCC 4401 as a potential probiotic for use in the food industry.

The Probiotic Properties and Safety of Limosilactobacillus mucosae NK41 and Bifidobacterium longum NK46

“…Lactate exists in two stereoisomeric forms, namely the L-form and D-form. Some strains of LAB, including Lactobacillus and Bifidobacterium genera, produce both D(−)-lactate and L(+)-lactate [10,49]. Since D(−)-lactate cannot be metabolized in the human gut, the excessive production and accumulation by gut bacteria could potentially lead to D-lactate acidosis and short bowel syndrome [50].…”

Screening of Lactobacillus from breast milk and infant feces and evaluation of their bile salt tolerance

“…In recent years, studies have found that breast milk is an important source of oral (Ruiz et al, 2019) and gastrointestinal bacteria in breastfed infants and this has an important impact on the early intestinal flora and the health of infants (McGuire and McGuire, 2015). With the maturity of strain isolation technology and culture conditions, a large number of probiotics, such as Lactobacillus rhamnosus (Li et al, 2020), L. plantarum, Bifidobacterium breve (Choi et al, 2021), and Bifidobacterium longum (Hikaru et al, 2020) have been isolated from breast milk and infant feces.…”

Genomic Characterization and Safety Assessment of Bifidobacterium breve BS2-PB3 as Functional Food

“…Bifidobacterium breve is notable for its antimicrobial properties against human pathogens, non-cytotoxic nature, and immunostimulatory abilities. This probiotic strain is commonly used in children as studies have demonstrated that B. breve in the GIT of infants could help reduce diarrhea occurrence, prevent allergic diseases, and protect infants from developing necrotising enterocolitis [ 6 - 10 ]. Building on its success in pediatric applications, there is a growing interest in exploring the potential use of B. breve for a wider population [ 11 , 12 ].…”