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Interaction of Bordetella bronchiseptica and Its Lipopolysaccharide with In Vitro Culture of Respiratory Nasal Epithelium.

Gallego C, Middleton AM, Martínez N, Romero S, Iregui C.

Vet Med Int. 2013;2013:347086.

Department of Veterinary Pathology, University of Applied and Environmental Sciences, Bogotá, Colombia.

 

Abstract

 

The nasal septa of fetal rabbits at 26 days of gestation were harvested by cesarean section of the does while under anesthesia and then exposed toBordetella bronchiseptica or its lipopolysaccharide (LPS) for periods of 2 and 4 hours. A total of 240 explants were used. The tissues were examined using the Hematoxylin & Eosin technique. Then, semithin sections (0.5 um) were stained with toluidine blue and examined with indirect immunoperoxidase (IPI) and lectin histochemistry. The most frequent and statistically significant findings were as follows: (1) cell death and increased goblet cell activity when exposed to bacteria and (2) cell death, cytoplasmic vacuolation and infiltration of polymorphonuclear leukocytes when exposed to LPS. The lesions induced by the bacterium were more severe than with LPS alone, except for the cytoplasmic vacuolation in epithelial cells. IPI stained the ciliated border of the epithelium with the bacterium more intensely, while LPS lectin histochemistry preferentially labeled the cytoplasm of goblet cell. These data indicate that B. bronchiseptica and its LPS may have an affinity for specific glycoproteins that would act as adhesion receptors in both locations.

 

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Additional Information:

In addition to the published paper {Interaction of Bordetella bronchiseptica and Its Lipopolysaccharide with In Vitro Culture of Respiratory Nasal Epithelium}, and using the same in vitro protocol, we obtained similar findings working with Pasteurella multocida and its lipopolysaccharide, another rabbit pathogen, which affects a broader spectrum of mammals and bird species (currently we are writing the manuscript). The purpose of these in vitro tissue cultures, actually, is to have an easy and more “real” model than cell cultures for different assays regarding potential preventive substances and measures against the infection by these two pathogens and in the future other respiratory tract pathogens. Accordingly, we have tested lectins- carbohydrates binding glycoproteins (submitted paper)- and also carbohydrates alone using these septum cultures for preventing the adhesion of P. multocida to the respiratory epithelium with promising success (work in progress).