The effect of calcium ions on adhesion and competitive exclusion of Lactobacillus ssp. and E. coli O138

Abstract

The adhesion abilities of 11 strains of Lactobacillus were determined in vitro using the IPEC-J2 cell line as a model system. Bacteria cultures included the probiotic strains L. rhamnosus GG, L. reuteri ATCC 55730, L. johnsonii NCC 533 and L. reuteri DSM 12246, and new isolates of Lactobacillus ssp. Adhesion was quantified by scintillation counting of radiolabelled bound bacteria. The highest adhesion of 38%, was determined for L. reuteri DSM 12246 followed by L. plantarum Q47 with an adhesion level of 24%. Other strains showed moderate to low binding of less than 16%. Competitive adhesion experiments on IPEC-J2 cells demonstrated that strongly adhesive strains, as L. reuteri DSM 12246 and L. plantarum Q47, significantly reduced the attachment of the less adhesive strains, such as L. rhamnosus GG and L. johnsonii NCC 533, both under condition of co-incubation and in displacement assays, indicating that bacteria may share the same binding sites for attachment to intestinal cells. Furthermore, it was revealed that calcium ions significantly increased the binding of tested lactobacilli to IPEC-J2 cells; and therefore, added calcium may be useful in enhancing the adhesion of normally weakly adhesive probiotic cultures. In contrast, no significant change in adhesion of lactobacilli was observed in the presence of Mg and Zn ions. Displacement assays performed with pathogenic E. coli O138 showed that all tested Lactobacillus strains reduced the attachment of E. coli O138 to IPEC-J2 by more than 2-fold both in the presence and the absence of calcium ions. The strains of Lactobacillus did not differ significantly in the extent of their inhibition of E. coli O138 adhesion, indicating that the reduced adhesion of E. coli O138 was due to steric hindrance of the binding sites rather than to specific interactions.