Resistance of Bordetella pertussis Phase I to Mucociliary Clearance by Rabbit Tracheal Mucous Membrane

Abstract

Cultivated mucous membrane of rabbit trachea was point inoculated with Bordetella pertussis phase I or III. Phase I (virulent) bacteria were found to be infective at the point-inoculated site, but phase III (avirulent) bacteria rarely showed such behavior. After inoculation, homogenized segments of mucous membrane were spread on plates. Large numbers of phase I bacteria were recovered from the inoculated segment; however, the laryngeal segment was the site of recovery of large numbers of phase III bacteria. The difference was not due to the ciliostatic bacterial toxin. After coinoculation with phase I and III bacteria, phase III bacteria were recognized at the laryngeal end, whereas phase I bacteria were recognized at the inoculated site. Therefore, the adherence of the bacteria to the ciliated epithelium is considered to be the most probable mechanism of the resistance to the mucociliary stream. Scanning electron microscopy facilitated visualization of phase I bacteria adhering to the inoculated site.

Scanning Electronmicroscopic Studies of Bordetella Bronchiseptica on the Rabbit Tracheal Mucosa

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PLATES VIII-XIIN previous studies Bordetella pertussis was shown to be resistant to the mucociliary clearance mechanism of the rabbit trachea (Matsuyama, 1977). Scanning electron microscopy readily demonstrated B.pertussis organisms associated with clusters of the cilia of the epithelial cells.

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“…Each explant was washed with 2 ml of saline, expelled through a 22-gauge needle at the rate of 0.6-0.8 ml/s to free the ciliated surfaces from mucus. Fixation and observation were as described previously (Matsuyama, 1977).…”

“…5). (Matsuyama, 1977), resulted in intimate interactions between bacteria and ciliated epithelial cells. On the mucous membrane infected with B. bronchiseptica most of the remaining cilia were entangled with large numbers of bacteria.…”

“…B. bronchiseptica infection usually started from the point-inoculated site on the ERTC, as in B. pertussis infection (Matsuyama, 1977). It seems possible that the interaction between cilia and organisms assists B. bronchiseptica in persisting in the respiratory tract.…”

An Assay of Bordetella Pertussis Adhesion to Tissue-Culture Cells

“…The ability of B. pertussis to adhere to cell surfaces has been proposed as one important factor associated with the virulence of the organism (Matsuyama, 1977;Sat0 et al, 1979). This property has been examined by various methods including haemagglutination tests (Keogh and North, 1948), microscopic observations of bacteria attached to tissue-culture cells (Holt, 1972;Sat0 et al, 1981), microscopic and viable counts of bacteria bound to rabbit, hamster or chick tracheal organ cultures (Iida and Ajiki, 1974;Collier, Peterson and Baseman, 1977;Matsuyama, 1977) and the assessment of bacterial adhesion to mouse lungs by radiolabelling techniques (Burns and Freer, 1982). However, attempts to quantify these methods are prone to difficulties with enumeration, contamination or uneven distribution of surface cell types, which may interfere with accurate and reproducible measurements of bacterial adhesion.…”