A fluorescence-based method for the detection of adhesive properties of lactic acid bacteria to Caco-2 cells.
OBJECTIVE
The ability of probiotic micro-organisms to adhere to the intestinal surface is regarded as a substantial advantage in terms of bacteria persistence in the gastrointestinal tract. The aim of the present study was the development of a method based on fluorescent staining of bacteria and subsequent spectrofluorimetric detection to quantify the adhesion of several strains of Lactobacillus and Bifidobacterium to Caco-2 cells.


RESULTS
Lactic acid bacteria strains were subjected to fluorescent staining using the viable probe carboxyfluorescein diacetate and subsequently incubated on Caco-2 monolayers. The adhesion of the micro-organisms was determined by spectrofluorimetry following the lysis of the attached bacterial cells and expressed as adhesion percentage. The values obtained for the micro-organisms tested ranged from 4% for Bifidobacterium infantis Bi1 to 10% for a Bifidobacterium mixture containing three different strains.


CONCLUSIONS
In the present study we successfully applied fluorescent labelling and fluorimetric detection to investigate the adhesive properties of some Lactobacillus and Bifidobacterium strains and a Bifidobacterium mixture to Caco-2 cells.


CONCLUSIONS
The results proved that fluorescent labelling is suitable for adhesion studies and provides a reliable and safer alternative to radioactive labelling.
Department of Public Health, University of Parma, Parma, Italy. mbianchi@nemo.unipr.it
M A
MA
Bianchi
D
D
Del Rio
N
N
Pellegrini
G
G
Sansebastiano
E
E
Neviani
F
F
Brighenti
D001422:Bacterial Adhesion; D001644:Bifidobacterium / Q000502:physiology*; D018938:Caco-2 Cells; D005452:Fluoresceins / Q000378:metabolism*; D006801:Humans; D007778:Lactobacillus / Q000502:physiology*; D008856:Microscopy, Fluorescence; D019936:Probiotics
D005452:Fluoresceins; C027780:carboxyfluoresceindiacetate