Effect of Bifidobacterium breve M-16V Supplementation on Fecal Bifidobacteria in Preterm Neonates - A Randomised Double Blind Placebo Controlled Trial

PLoS ONE

Mar 2014

DOI: 10.1371/journal.pone.0089511

Citations:121

Influential Citations:3

Interventional (Human) Studies

Enhanced Details

Methods

Randomized, placebo-controlled trial in preterm neonates (<33 weeks gestation) ready to commence enteral feeds or fed within 12 hours. 159 neonates enrolled (79 probiotic, 80 placebo); primary outcome: fecal B. breve counts by quantitative PCR; secondary outcomes included total fecal bifidobacteria, NEC ≥ Stage II, all-cause death, time to reach full enteral feeds and late-onset sepsis; intention-to-treat analysis; allocation masked for stool analyses.

Intervention

Bifidobacterium breve M-16V. Dose: 3.6e9 CFU per day (1.5 mL of reconstituted solution) given as a single daily dose via an orogastric feeding tube; started when enteral feeds were possible and continued until corrected age 37 weeks; for neonates gestation ≤27 weeks, start with 1.56e9 CFU per day until milk feeds reach 50 mL/kg/day, then increase to 3.6e9 CFU per day.

Results

After 3 weeks, detectable B. breve increased to 91% (67/74) in probiotic vs 38% (25/66) in placebo (p<0.001). Median B. breve counts rose from below detection (<4.7 log CFU/g) at baseline to 8.6 log CFU/g in the probiotic group, while remaining <4.7 log CFU/g in placebo (p<0.001). No adverse events including probiotic sepsis; NEC Stage II occurred in 1 placebo neonate; no deaths. Conclusion: B. breve M-16V is a suitable probiotic strain for routine use in preterm neonates.

Limitations

Not powered to detect NEC or all-cause mortality; sample size adjustments and stool collection challenges; potential cross-contamination between groups; inability to definitively confirm M-16V in stool by PCR; single-center study with limited generalizability.

Abstract

Background Probiotic supplementation significantly reduces the risk of necrotising enterocolitis (NEC) and all cause mortality in preterm neonates. Independent quality assessment is important before introducing routine probiotic supplementation in this cohort. Aim To assess product quality, and confirm that Bifidobacterium breve (B. breve) M-16V supplementation will increase fecal B. breve counts without adverse effects. Methods and Participants Strain identity (16S rRNA gene sequencing), viability over 2 year shelf-life were confirmed, and microbial contamination of the product was ruled out. In a controlled trial preterm neonates (Gestation <33 weeks) ready to commence or on feeds for <12 hours were randomly allocated to either B. breve M-16V (3×109 cfu/day) or placebo (dextrin) supplementation until the corrected age 37 weeks. Stool samples were collected before (S1) and after 3 weeks of supplementation (S2) for studying fecal B. breve levels using quantitative PCR (Primary outcome). Secondary outcomes included total fecal bifidobacteria and NEC≥Stage II. Categorical and continuous outcomes were analysed using Chi-square and Mann-Whitney tests, and McNemar and Wilcoxon signed-rank tests for paired comparisons. Results A total of 159 neonates (Probiotic: 79, Placebo: 80) were enrolled. Maternal and neonatal demographic characteristics were comparable between the groups. The proportion of neonates with detectable B. breve increased significantly post intervention: Placebo: [S1:2/66 (3%), S2: 25/66 (38%), p<0.001] Probiotic: [S1: 29/74 (40%), S2: 67/74 (91%), p<0.001]. Median S1 B. breve counts in both groups were below detection (<4.7 log cells.g−1), increasing significantly in S2 for the probiotic group (log 8.6) while remaining <4.7 log in the control group (p<0.001). There were no adverse effects including probiotic sepsis and no deaths. NEC≥Stage II occurred in only 1 neonate (placebo group). Conclusion B. breve M-16V is a suitable probiotic strain for routine use in preterm neonates. Trial Registration Australia New Zealand Clinical Trial Registry ACTRN 12609000374268 show less