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. 2016 Jul;30(4):1204-9.
doi: 10.1111/jvim.14366.

Quantitative PCR and Cytology of Bronchoalveolar Lavage Fluid in Dogs with Bordetella bronchiseptica Infection

A M Canonne  1 , F Billen  1 , C Tual  1 , E Ramery  1 , E Roels  1 , I Peters  2 , C Clercx  1
Affiliations

Quantitative PCR and Cytology of Bronchoalveolar Lavage Fluid in Dogs with Bordetella bronchiseptica Infection

A M Canonne et al. J Vet Intern Med. 2016 Jul.

Abstract

Background: The use of quantitative PCR (qPCR) for detection of Bordetella bronchiseptica in bronchoalveolar lavage fluid (BALF) and demonstration of bacteria adhering to ciliated epithelial cells in BALF or bronchial brushing fluid (BBF) has not been assessed in a series of affected dogs. Coinfections can worsen the clinical severity in bordetellosis, but the specific association with Mycoplasma cynos has not been evaluated.

Objectives: To assess the utility of culture, qPCR and cytologic examination of cytospin preparations in the diagnosis of bordetellosis in dogs and the influence of coinfection by M. cynos on disease severity.

Animals: Twenty-four referred dogs with B. bronchiseptica infection and 10 healthy dogs.

Methods: Retrospective case series. qPCR (B. bronchiseptica and M. cynos) and culture results from BALF were recorded. Cytospin preparations from BALF and BBF were reviewed. qPCR on BALF from 10 healthy dogs were used as negative control.

Results: The BALF culture and qPCR detected B. bronchiseptica in 14/24 and 18/18 dogs, respectively. Coccobacilli were found adhering to ciliated epithelial cells in 20 of the 21 BALF cytologic preparations where epithelial cells were found, and 2/3 BBF cytologic preparations. Quantitative PCR detected a low level of B. bronchiseptica in one healthy dog. The frequency of detection of M. cynos was not significantly different in B. bronchiseptica (9/17 dogs) compared with healthy dogs (2/10 dogs) (P = .09).

Conclusion and clinical importance: Quantitative PCR detection of B. bronchiseptica in BALF appears to be a useful diagnostic tool. Cytologic examination of BALF or BBF, when positive, allows a rapid and reliable diagnosis.

Keywords: Bronchial brushing fluid; Clinical pathology; Cytology; Mycoplasma cynos; qPCR.

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Figures

Figure 1
Figure 1
Bronchoscopic view of tracheobronchic bifurcation showing congested mucosa and thick and white‐yellow material (Dog n°3, sharpei, female, 6 months old).
Figure 2
Figure 2
Cytological detection of pleiomorphic cocci or coccobacilli adhering to the cilia of the epithelial cells of BALF (May‐Grünwald‐Giemsa stain, ×400 magnification, Dog n°8, Yorkshire Terrier, female, 5 months old).
See this image and copyright information in PMC

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