title: In vitro adherence properties of Lactobacillus rhamnosus DR20 and Bifidobacterium lactis DR10 strains and their antagonistic activity against an enterotoxigenic Escherichia coli.
abstract: Adhesion and colonisation properties of three probiotic strains namely, Lactobacillus rhamnosus DR20, L. acidophilus HN017, and Bifidobacterium lactis DR10, were determined in vitro using the differentiated human intestinal cell-lines including HT-29, Caco-2, and HT29-MTX, and compared with properties of L. acidophilus LA-1 and L. rhamnosus GG (two commercial probiotic strains). Two independent methods were employed to quantitate the "adhesiveness" of each strain. In the first method, the bacteria adhered to human cells were detected by Gram staining and counted in different fields under a microscope. Bacteria were also radio-labelled and extent of adhesion determined by scintillation counting. All three strains showed strong adhesion with the human intestinal cell lines in vitro. Adhesion indices of the three strains to two cell lines, i.e. HT-29, and Caco-2 varied between 99 +/- 17 and 219 +/- 36. With mucus-secreting cell-line HT29-MTX, the adhesion indices of all the strains were 2-3 times higher. The adhesion indices of L. acidophilus LA-1 and L. rhamnosus GG were comparable to the other three probiotic strains. We also investigated the inhibitory effect of adhering strains against the intestinal cell monolayer colonization by a known enterotoxigenic strain of Escherichia coli (strain O157:H7). Pre-treatment of E. coli O157:H7 with 2.5-fold concentrated cell-free culture supernatants from L. acidophilus HN017, L. rhamnosus DR20 and B. lactis DR10 reduced the culturable E. coli numbers on TSB plates and also reduced the invasiveness and cell association characteristics of this toxic strain. The inhibitory molecules secreted into the spent media by these strains were partially affected by treatments with lactate dehydrogenase, trypsin and proteinase K suggesting that overall inhibition may be due to a synergistic action of lactic acid and proteinaceous substances.
authors.affiliation: New Zealand Dairy Research Institute, Palmerston North. pramod.gopal@nzdri.org.nz
authors.forename: P K
authors.initials: PK
authors.lastname: Gopal
authors.forename: J
authors.initials: J
authors.lastname: Prasad
authors.forename: J
authors.initials: J
authors.lastname: Smart
authors.forename: H S
authors.initials: HS
authors.lastname: Gill
mesh_terms: D001422:Bacterial Adhesion / Q000502:physiology*; D001644:Bifidobacterium / Q000502:physiology*; D001667:Binding, Competitive; D018938:Caco-2 Cells; D002460:Cell Line; D015169:Colony Count, Microbial; D019453:Escherichia coli O157 / Q000254:growth & development*; D005840:Gentian Violet; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism / Q000382:microbiology; D007778:Lactobacillus / Q000502:physiology*; D008855:Microscopy, Electron, Scanning; D010619:Phenazines; D019936:Probiotics / Q000378:metabolism*; D012588:Scintillation Counting
chemical_list: C035863:Gram's stain; D010619:Phenazines; D005840:Gentian Violet