Adhesion of probiotic micro-organisms to intestinal mucus

doi:10.1016/S0958-6946(99)00132-6

International Dairy Journal

Volume 9, Issue 9, September 1999, Pages 623-630

https://doi.org/10.1016/S0958-6946(99)00132-6 Get rights and content

Abstract

For many of the proposed health effects of probiotic micro-organisms it is desirable that the organism at least transiently colonises the gastro-intestinal tract. Interaction with the intestinal mucosa may enhance the possibility for colonisation. In this study, we investigated the adhesion to human intestinal mucus of a human faecal isolate, probiotic, dairy and type culture strains. A significant variation in adhesion between the strains was observed; ranging from 3% (Lactobacillus casei 01) to 43% (L. rhamnosus GG) adhesion of the applied cells. Interestingly, two of the tested dairy strains; L. bulgaricus and Lactococcus lactis ssp. cremoris, were found to adhere well. Adhered micro-organisms were found to be tightly bound to the immobilised mucus. No correlation was observed between cell surface hydrophobicity and adhesive ability of the strains. The observed differences in adhesion may help to explain why certain strains are excreted longer than others after cessation of their administration and have different health effects.

Introduction

Lactic acid bacteria and other probiotic micro-organisms have many documented health effects (Salminen, Isolauri & Salminen, 1996). When selecting probiotic micro-organisms with beneficial health effects for the host, many criteria have to be met. Among these criteria is the ability to adhere to host tissue (Ouwehand, Kirjavainen, Shortt & Salminen, 1999a). Adhesion to the intestinal mucosa is considered a prerequisite for successful colonisation (Finlay & Falkow, 1997; Doig & Trust, 1993) and is important for immune modulation by the probiotics (Ouwehand et al., 1999a; Famularo, Moretti, Marcellini & De Simone, 1997).

Adhesion of micro-organisms is generally tested with mono-layers of intestinal tissue culture cells. Although this is a valuable method for assessing the adhesive abilities of bacteria to the intestinal mucosa, it does not take into account possible adhesion to the mucus layer covering the epithelial cells in the intestine. The mucus layer has a dual role. It serves as a barrier for adhesion of certain micro-organisms to the underlying epithelium (Freter, 1984; Carlstedt-Duke, 1989), while at the same time it can provide a habitat for the adhesion of others (Nielsen, Schlundt, Gunvig & Jacobsen, 1994; van der Waaij, Limburg, Mesander & van der Waaij, 1996). In the present study, we describe the use of intestinal mucus as a substratum for adhesion studies. The mucus was isolated from subjects of different age to determine the effect of age on adhesion of probiotic bacteria.

It has been shown previously that the composition of the intestinal Lactobacillus population changes in time (Mitsuoka & Hayakawa, 1972; Kimura, McCartney, McConnell & Tanock, 1997). The age of the subject may therefore play a role in the ability of the probiotic strain to adhere and colonise the intestinal mucosa. This in turn can influence the efficacy of the probiotic preparation used. Knowledge about the adhesive properties of the probiotic organisms can therefore give information about its possibility to colonise and modulate the immune system.

The surface hydrophobicity of the tested micro-organisms has been determined in order to test for a possible correlation between this physico-chemical property and the ability to adhere to intestinal mucus as suggested by Wadström, Andersson, Sydow, Axelsson, Lindgren and Gullmar (1987). This would provide an easy and quick method to screen for potential adhesive probiotic strains.

Section snippets

Micro-organisms and growth conditions

The micro-organisms used and their growth conditions are listed in Table 1. The bacteria were a generous gift from Dr. M. Saxelin (Valio Dairies Ltd, Helsinki, Finland). Saccharomyces boulardii was isolated from Precosa^® (Astra, Södertälje, Sweden). All media (3 ml) contained 30 μl of methyl-1,2[ $^{3} H$ ]thymidine (6.7 Ci mmol⁻¹; Nen products) to metabolically label the bacteria. Bacteria were harvested by centrifugation (1000×g), washed in HEPES (N-2-hydroxy-ethylpiperazine-N′-2-ethanesulfonic

Mucus characterisation

The results of the mucus characterisation are shown in Table 2. Mucus prepared from 6 month old and adult subjects did not contain detectable amounts of sulphate (<0.05%). The results show that in all mucus preparations little or no H-antigens were present. All mucus preparations show high A-antigen activity and, except for mucus isolated from 6 month old infants, also relatively high B-antigen activity.

The results of the adhesion of the tested probiotic bacteria to intestinal mucus are shown

Discussion

The ability of micro-organisms to adhere is often considered one of the main selection criteria for potential probiotics (Ouwehand et al., 1999a). Adhesion to the intestinal mucosa is thought to be an important property for colonisation by preventing wash-out (Wadström, 1988), especially in the small intestine where flow rates are relatively high (Sanford, 1992). Adhesion to the intestinal mucosa has also been suggested to enhance the ability to stimulate the immune system (Ouwehand et al.,

Acknowledgements

The authors wish to thank P. Gröndal (Finnish Red Cross, Turku) for providing erythrocytes for the blood-group antigen determination. Financial support was obtained from the Academy of Finland.

References (46)

P.S. Cohen et al.
Bacterial adhesion to and penetration of intestinal mucus in vitro
Methods in Enzymology
(1995)
P. Doig et al.
Methodological approaches of assessing microbial binding to extracellular matrix components
Journal of Microbiological Methods
(1993)
P.V. Kirjavainen et al.
The ability of probiotic bacteria to bind to human intestinal mucus
FEMS Microbiology Letters
(1998)
O.H. Lowry et al.
Protein measurement with the folin phenol reagent
Journal of Biological Chemistry
(1951)
R.S. Miller et al.
Mucus degradation in human colon ecosystems. Fecal population densities of mucus-degrading bacteria estimated by a `most probable number’ method
Gastroenterology
(1981)
A.C. Ouwehand et al.
Probioticsmechanisms and established effects
International Dairy Journal
(1999)
A.C. Ouwehand et al.
The normal faecal microflora does not affect the adhesion of probiotic bacteria in vitro
FEMS Microbiology Letters
(1999)
P.T.J. Willemsen et al.
Age and serotype dependent binding of K88 fimbriae to porcine intestinal receptors
Microbiology and Pathology
(1992)
S. Ashkenazi et al.
The effect of postnatal age on the adherence of Shigella flexneri, Escherichia coli 0124, and E. coli 0128 to guinea pig intestinal cells
Pediatric Research
(1984)
S. Ashkenazi et al.
The effect of postnatal age on the adherence of enterohemorrhagic Escherichia coli to rabit intestinal cells
Pediatric Research
(1991)

M.F. Bernet et al.

Lactobacillus acidophilus LA 1 binds to cultured human intestinal cell lines and inhibits cell attachment and cell invasion by enterovirulent bacteria

Gut

(1994)

Bianchi-Salvadori, B. (1986). Intestinal microflora: the role of yoghurt in the equilibrium of the gut ecosystem....

Brassart, D., Neeser, J.-R., Michetti, P., Servin, A. L. (1994). The selection of dairy bacterial strains with...

Carlstedt-Duke, B. (1989). The normal microflora and mucin. In R. Grubb, T. Midtvedt, & E. Norin, The regulatory and...

M.-H. Coconnier et al.

Protein-mediated adhesion of Lactobacillus acidophilus BG2FO4 on human enterocyte and mucus-secreting cell lins in culture

Applied and Environmental Microbiology

(1992)

P.L. Conway et al.

Presence of K88-specific receptors in porcine ileal mucus is age dependent

Infection and Immunity

(1990)

E.A. Dean

Comparison of receptors for 987P pili of enterotoxigenic Escherichia coli in the small intestines of neonatal and older pigs

Infection and Immunity

(1990)

M. Dubois et al.

Colorimetric method for determination of sugars and related substances

Analytical Chemistry

(1956)

A.M.A. El Monem et al.

The effect of postnatal development on the adherence of nonfimbriated and fimbriated Salmonella typhimurium to isolated small intestinal enterocytes

Pediatric Research

(1988)

Elliot, S. N., Buret, A., McKnight, W., Miller, M. J. S., & Wallace, J. L. (1998). Bacteria rapidly colonize and...

S. Elo et al.

Attachement of Lactobacillus casei strain GG to human colon carcinoma cell line Caco-2comparison with other dairy strains

Letters in Applied Microbiology

(1991)

G. Famularo et al.

Stimulation of immunity by probiotics

B.B. Finlay et al.

Common themes in microbial pathogenicity revisited

Microbiology and Molelcular Biology Reviews

(1997)

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Adhesion of probiotic micro-organisms to intestinal mucus

Abstract

Introduction

Section snippets

Micro-organisms and growth conditions

Mucus characterisation

Discussion

Acknowledgements

Methods in Enzymology

Journal of Microbiological Methods

FEMS Microbiology Letters

Journal of Biological Chemistry

Gastroenterology

International Dairy Journal

FEMS Microbiology Letters

Microbiology and Pathology

The effect of postnatal age on the adherence of Shigella flexneri, Escherichia coli 0124, and E. coli 0128 to guinea pig intestinal cells

Pediatric Research

The effect of postnatal age on the adherence of enterohemorrhagic Escherichia coli to rabit intestinal cells

Pediatric Research

Lactobacillus acidophilus LA 1 binds to cultured human intestinal cell lines and inhibits cell attachment and cell invasion by enterovirulent bacteria

Gut

Protein-mediated adhesion of Lactobacillus acidophilus BG2FO4 on human enterocyte and mucus-secreting cell lins in culture

Applied and Environmental Microbiology

Presence of K88-specific receptors in porcine ileal mucus is age dependent

Infection and Immunity

Comparison of receptors for 987P pili of enterotoxigenic Escherichia coli in the small intestines of neonatal and older pigs

Infection and Immunity

Colorimetric method for determination of sugars and related substances

Analytical Chemistry

The effect of postnatal development on the adherence of nonfimbriated and fimbriated Salmonella typhimurium to isolated small intestinal enterocytes

Pediatric Research

Attachement of Lactobacillus casei strain GG to human colon carcinoma cell line Caco-2comparison with other dairy strains

Letters in Applied Microbiology

Stimulation of immunity by probiotics

Common themes in microbial pathogenicity revisited

Microbiology and Molelcular Biology Reviews