The frog palate mucosa as a model for studying bacterial adhesion to mucus-coated respiratory epithelium

Abstract

Most of the methods proposed to quantify bacterial adherence to respiratory mucosa differ mainly from in vivo conditions in the absence of the mucus blanket and in the exposure of the sub-mucosal connective tissue (SMCT) to the micro-organisms. We propose the frog palate as a model to study bacterial adhesion to the respiratory mucosa, with a system which allows the mucus to be preserved and the bacterial adhesion to be quantified in a standardized mucosal area, where mucociliary transport is still active. In order to evaluate the role of respiratory mucus in bacteria-mucosa interaction, we compared the adhesion of radiolabelled pneumococci to 12 mucus-coated and 10 non-mucus-coated frog palate mucosae. The presence or absence of mucus was controlled by scanning electron microscopy (SEM). After a 10 min incubation period, the bacterial adhesion to mucus-coated palate mucosa was five times greater ( P < 0·01) than that to uncoated mucosa. By SEM, bacteria were never seen attached to ciliated cells but could be detected on small areas where mucus was not totally eliminated. Even after a 120 min contact of bacteria to uncoated mucosa, bacterial adhesion remained only half that to mucus-coated epithelium. In order to ascertain whether the exposure of the SMCT represented a means of attraction to bacteria, we incubated the frog palate mucosa face-down with radiolabelled Pseudomonas aeruginosa. As much as 44 per cent of added bacteria adhered to exposed SMCT and, by SEM, numerous micro-organisms were seen attached to connective tissue. In contrast, only a few bacteria were observed adhering to the mucosa, mainly to granules of mucus.