Selective method for identification and quantification of Bifidobacterium animalis subspecies lactis BB-12 (BB-12) from the gastrointestinal tract of healthy volunteers ingesting a combination probiotic of BB-12 and Lactobacillus rhamnosus GG

J Appl Microbiol. 2017 May;122(5):1321-1332. doi: 10.1111/jam.13436. Epub 2017 Apr 4.

Abstract

Aim: To develop a novel validated method for the isolation of Bifidobacterium animalis ssp. lactis BB-12 (BB-12) from faecal specimens and apply it to studies of BB-12 and Lactobacillus rhamnosus GG (LGG) recovered from the healthy human gastrointestinal (GI) tract.

Methods and results: A novel method for isolating and enumerating BB-12 was developed based on its morphologic features of growth on tetracycline-containing agar. The method identified BB-12 correctly from spiked stool close to 100% of the time as validated by PCR confirmation of identity, and resulted in 97-104% recovery of BB-12. The method was then applied in a study of the recovery of BB-12 and LGG from the GI tract of healthy humans consuming ProNutrients® Probiotic powder sachet containing BB-12 and LGG. Viable BB-12 and LGG were recovered from stool after 21 days of probiotic ingestion compared to baseline. In contrast, no organisms were recovered 21 days after baseline in the nonsupplemented control group.

Conclusions: We demonstrated recovery of viable BB-12, using a validated novel method specific for the isolation of BB-12, and LGG from the GI tract of healthy humans who consumed the probiotic supplement.

Significance and impact of the study: This method will enable more detailed and specific studies of BB-12 in probiotic supplements, including when in combination with LGG.

Keywords: Lactobacillus; enumeration; intestinal microbiology; isolation; probiotics.

Publication types

  • Randomized Controlled Trial

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • Anti-Bacterial Agents
  • Bifidobacterium animalis / classification
  • Bifidobacterium animalis / genetics
  • Bifidobacterium animalis / isolation & purification*
  • Bifidobacterium animalis / physiology
  • Dietary Supplements
  • Feces / microbiology
  • Female
  • Gastrointestinal Tract / microbiology*
  • Healthy Volunteers
  • Humans
  • Lacticaseibacillus rhamnosus / genetics
  • Lacticaseibacillus rhamnosus / isolation & purification
  • Lacticaseibacillus rhamnosus / physiology*
  • Male
  • Middle Aged
  • Probiotics / administration & dosage*
  • Tetracycline
  • Young Adult

Substances

  • Anti-Bacterial Agents
  • Tetracycline