Exposing a Synthetic Oral Bacterial Community to Simulated Phases of Digestion

Abstract

Source: Arroyo, M. C., et al. Assessing the Viability of a Synthetic Bacterial Consortium on the In Vitro Gut Host-microbe Interface. J. Vis. Exp. (2018)

This video demonstrates the viability assessment of a synthetic oral microbial community during simulated oral, gastric, and intestinal digestion. Bacterial survival is quantified after each phase, revealing a progressive decline due to digestive stress.

Protocol

1. Bacterial Survival Following In Vitro Gastrointestinal Transit Conditions

NOTE: Prepare simulated digestion fluids with the modifications described below. Prepare all the dilutions with ultrapure water. Filter-sterilize all solutions through a 0.22 µm filter and perform the digestion procedure under sterile conditions.

1. Oral digestion:
   1. Place 3 mL of the bacterial community in a 50 mL sterile tube, mix with 3 mL of simulated saliva fluid (SSF), containing in millimoles per liter (mmol L⁻¹): Potassium chloride (KCl), 15.1; Mon....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Mucin from porcine stomach Type II	Sigma-Aldrich	M2378
PBS	Gibco	14190250
Millipore Milli-Q academic, ultra pure water system	Millipore, Merck KGaA	-
InnOva 4080 incubator shaker	New Brunswick Scientific	8261-30-1007
Nuclease-free water	Serva Electrophoresis	28539010
BD Accuri C6 flow cytometer	BD Biosciences	653118
InnuPREP PCRpure kit	Analytik Jena	845-KS-5010250	PCR purification kit