Cécile Clercx

CCDC39 is required for assembly of inner dynein arms and the dynein regulatory complex and for normal ciliary motility in humans and dogs

Primary ciliary dyskinesia (PCD) is an inherited disorder characterized by recurrent infections of the upper and lower respiratory tract, reduced fertility in males and situs inversus in about 50% of affected individuals (Kartagener syndrome). It is caused by motility defects in the respiratory cilia that are responsible for airway clearance, the flagella that propel sperm cells and the nodal monocilia that determine left-right asymmetry. Recessive mutations that cause PCD have been identified in genes encoding components of the outer dynein arms, radial spokes and cytoplasmic pre-assembly factors of axonemal dyneins, but these mutations account for only about 50% of cases of PCD. We exploited the unique properties of dog populations to positionally clone a new PCD gene, CCDC39. We found that loss-of-function mutations in the human ortholog underlie a substantial fraction of PCD cases with axonemal disorganization and abnormal ciliary beating. Functional analyses indicated that CCDC39 localizes to ciliary axonemes and is essential for assembly of inner dynein arms and the dynein regulatory complex.

Eosinophilic bronchopneumopathy in dogs

Eosinophilic bronchopneumopathy was diagnosed in 23 young dogs. Clinical signs included cough, gagging, and retching in all dogs, dyspnea in 21 dogs (91%), and nasal discharge in 12 dogs (52%). The most common radiographic findings were a moderate to severe bronchointerstitial pattern (68%, 13 of 19 dogs). Bronchoscopic findings included the presence of abundant yellow-green mucus or mucopurulent material (70%, 16 of 23 dogs) and severe mucosal thickening with an irregular or polypoid appearance (52%, 12 of 23 dogs), with partial airway closure during expiration in 3 dogs (13%). Peripheral blood eosinophilia was noted in 14 of 23 dogs (61%). Inflammatory cells in brush or bronchoalveolar lavage fluid cytologic preparations comprised more than 50% eosinophils in 14 of 23 dogs (61%), and 20-50% eosinophils in 6 dogs (26%). Eosinophilic infiltration of the bronchial mucosa was observed in biopsies from 19 dogs and was graded as mild (37%, 7 dogs), moderate (32%, 6 dogs), or severe (32%, 6 dogs). The mean serum immunoglobulin A concentration was almost double that of a population of 20 healthy dogs of various breeds. Oral glucocorticoids were administered on alternate days with progressive tapering of the dose; the dosage at maintenance varied between 0.1 and 1.0 mg/kg every other day. No relationship was found between the duration of clinical signs and the maintenance dosage or the cytologic and histopathologic grades.

Quantitative Bacterial Cultures and Cytological Examination of Bronchoalveolar Lavage Specimens in Dogs

Cytology and quantitative bacterial cultures of lower respiratory tract secretions are widely used in human medicine to differentiate airway infection from simple bacterial colonization. A retrospective study was conducted to determine the usefulness of quantitative aerobic cultures and Gram stain intracellular bacteria counts from bronchoalveolar lavage (BAL) specimens in dogs in diagnosing lower respiratory tract infection (LRTI) and to determine whether chronic bronchitis is associated with marked bacterial growth in dogs. The threshold determined to define clinically relevant bacterial growth was 1.7 x 10(3) colony-forming units per milliliter of BAL fluid. We used this threshold and found that diagnostic sensitivity and specificity were 86% and 100%, respectively. With a threshold for infection of >2 intracellular bacteria observed in any of 50 fields, microscopic examination of Gram stain BAL preparations had a sensitivity of 71% and a specificity of 97% in establishing LRTI. There was a high correlation between bacterial morphology on BAL Gram stain and bacterial cultures. Combining the results of intracellular bacteria counts from the BAL Gram stain with those from the quantitative cultures, the sensitivity in diagnosing LRTI was 87% and the specificity was 97%. BAL quantitative cultures as well as quantitating intracellular bacteria on Gram stain BAL cytology were revealed to be useful in identifying LRTI in dogs. Chronic bronchitis does not appear to be associated with marked bacterial growth in dogs.

Clinical, bronchoscopic, histopathologic, diagnostic imaging, and arterial oxygenation findings in West Highland White Terriers with idiopathic pulmonary fibrosis.

BACKGROUND Idiopathic pulmonary fibrosis (IPF) is a chronic, interstitial lung disease primarily affecting West Highland White Terriers (WHWTs). OBJECTIVE To describe the clinicopathological and diagnostic imaging features in WHWTs with IPF. ANIMALS Twelve WHWTs with IPF and 14 healthy control WHWTs. METHOD Prospective study. Clinical signs and findings of physical examination, blood and arterial blood gas analyses, radiography, high-resolution computed tomography (HRCT), bronchoscopy and bronchoalveolar lavage (BAL) of IPF dogs were obtained and compared with controls. Histopathologic changes in IPF dogs were evaluated. RESULTS Mean partial pressure of oxygen was significantly lower in IPF (mean ± SD, 65.5 ± 15.4 mmHg) than in controls (99.1 ± 7.8 mmHg, P<.001). The alveolar-arterial oxygen gradient was significantly higher in IPF (50.1 ± 17.3 mmHg) than in controls (17.5 ± 4.9 mmHg, P<.001). In HRCT, ground glass opacity (GGO) was detected in all IPF dogs, traction bronchiectasis in 4, and honeycombing in 1. Bronchoscopic airway changes were noted in all IPF dogs. On BAL fluid (BALF) cytology, the total cell count (TCC) was higher in IPF dogs, and the numbers but not the percentages of macrophages, neutrophils, and mast cells were increased. On histopathology, multifocal or diffuse interstitial fibrosis, type II pneumocyte hyperplasia, prominent intraalveolar macrophages, distortion of alveolar architecture, and emphysematous change were detected. CONCLUSION AND CLINICAL IMPORTANCE IPF causes substantial hypoxemia. In HRCT, GGO is a consistent finding. IPF dogs have concurrent airway changes and an increase in BALF TCC.

Description of original endoscopic findings and respiratory functional assessment using barometric whole-body plethysmography in dogs suffering from brachycephalic airway obstruction syndrome

The clinical features of brachycephalic airway obstructive disease in 11 brachycephalic dogs are described in this study. The respiratory strategy was assessed before (n=11) and after (n=6) surgery using barometric whole-body plethysmography (BWBP), with the relationship between BWBP variables and the severity of the clinical signs assessed by the use of a respiratory score based on clinical, radiographic and endoscopic findings. Partial collapse of the left main bronchus was a common finding not previously described as part of the brachycephalic airway obstructive disease syndrome. Epiglottic cysts, laryngeal granulomas and nasopharyngeal turbinates in English Bulldogs were other previously unreported findings. No significant correlation between the respiratory score and any of the BWBP variables was detected. Compared to healthy dogs, brachycephalic dogs had a significantly lower Te/Ti ratio (expiratory time over inspiratory time), peak inspiratory flow (PIF) per kg bodyweight (BW), significantly higher peak expiratory flow (PEF) per kgBW, PEF/PIF, and enhanced pause. These variations are compatible with upper airway obstructions primarily in the extrathoracic airways. Following surgery, a significant decrease in PEF/PIF was detected. The study showed that BWBP could be used to characterise the respiratory strategy in brachycephalic dogs before and after surgery.

Inhaled fluticasone reduces bronchial responsiveness and airway inflammation in cats with mild chronic bronchitis.

This study investigated the effect of inhaled fluticasone on lower airway inflammation and bronchial responsiveness (BR) to inhaled carbachol in cats with very mild, chronic bronchitis (n=5) that were compared with healthy cats serving as controls (n=6). Chest radiographs, BR tests performed non-invasively by barometric whole body plethysmography (BWBP) and bronchoalveolar lavage (BAL) were performed before and after treatment. BR was quantified by calculating the concentration of carbachol inducing bronchoconstriction (C-Penh300%), defined as a 300% increase of baseline Penh, an index of bronchoconstriction obtained by BWBP. BAL fluid was analyzed cytologically and the oxidant marker 8-iso-PGF2α was determined. At test 1, healthy cats and cats with bronchitis were untreated, whereas for test 2 inhalant fluticasone (250 μg once daily) was administrated for 2 consecutive weeks to cats with bronchitis. Control cats remained untreated. Inhaled fluticasone induced a significant increase in C-Penh300% and a significant decrease of BAL fluid total cells, macrophages, neutrophils and 8-iso-PGF2α in cats with bronchitis, whilst untreated control cats did not show significant changes over time. This study shows that a 2-week fluticasone treatment significantly reduced lower airway inflammation in very mild bronchitis. BR could be successfully monitored in cats using BWPB and decreased significantly in response to inhaled fluticasone. 8-Iso-PGF2α in BAL fluid was responsive to treatment and appeared as a sensitive biomarker of lower airway inflammation in cats.

Bronchodilators in bronchoscopy-induced airflow limitation in allergen-sensitized cats.

This study investigated the effect of bronchoscopy and bronchoalveolar lavage (BAL) on respiratory function, determined by barometric whole-body plethysmography (BWBP), of healthy and allergen-sensitized cats. Furthermore, the efficacy of inhaled bronchodilators in preventing changes in respiratory function was determined. For test 1, 18 healthy experimental cats were investigated on day 1 by BWBP. On day 2, the cats underwent BWBP after sedation (medetomidine), after anesthesia induction (propofol), and after bronchoscopy and BAL. Enhanced pause (Penh) was significantly increased after bronchoscopy and BAL (1.64 +/- 0.17 versus 1.23 +/- 0.07, P < .05). For test 2, 6 cats were sensitized to ovalbumin (OVA), 6 cats were sensitized to Ascaris suum (AS), and 6 cats served as controls. On day 0, OVA- and AS-sensitized cats underwent an inhaled allergen challenge, whereas controls were exposed to saline. On days 1 and 2, the same protocol as described for test 1 was repeated. Post-BAL Penh of the AS-sensitized cats was significantly higher than at test 1 (2.28 +/- 0.22 versus 1.69 +/- 0.33, P < .05) and was correlated with BAL fluid neutrophil count (r = 0.55, P < .05). During tests 3, 4, and 5, the same protocol as used for test 2 was applied to each cat group, with the animals being randomly treated before sedation with inhaled salbutamol (200 microg), ipratropium bromide (40 microg), or a combination of both (200 + 40 microg). Post-BAL Penh of the AS-sensitized group was significantly decreased after the salbutamol + ipratropium bromide treatment (1.56 +/- 0.18 versus 2.28 +/- 0.22, P < .05). This study suggests that bronchoscopy and BAL induce airflow limitation in cats, which is more severe in the presence of lower airway inflammation. Inhaled salbutamol + ipratropium bromide reduce BAL-induced bronchoconstriction in AS-challenged cats and might be recommended as preventive treatment of asthmatic cats undergoing bronchoscopy.

A retrospective study of non-specific rhinitis in 22 cats and the value of nasal cytology and histopathology

Case records from 40 cats subjected to rhinoscopic examination for investigation of chronic nasal disease were reviewed. Cases in which no specific underlying cause (eg neoplasia) was detected were further selected for detailed retrospective study. In these 22 cats (55% of the initial population), a final diagnosis of non-specific chronic nasal disease was made. The radiographic, rhinoscopic, cytological and histopathological findings were reviewed. Mucosal biopsy specimens were obtained in 20 cases. Despite clinical signs of more than 4 weeks duration, histopathology indicated acute inflammation in four cases. Two cases had chronic lymphoplasmacytic inflammation and 14 had mixed (lymphoplasmacytic and neutrophilic) inflammation. Specimens for cytology were obtained from 17 cases by brush sampling. Three of these samples were not diagnostic due to the poor quality of the slides; one showed normal cytology. Acute inflammation was diagnosed by cytology (n=11) more commonly than chronic (n=1) or mixed inflammation (n=1). Concurrent samples, of quality suitable for both histopathological and cytological interpretation, were collected from 12 cases only. Cytological results were in agreement with the histological results in 25% of these cases, the main discrepancy being the nature of the dominant inflammatory cell type. Therefore cytology does not appear to be a reliable means for detection of chronic inflammation. Further studies are needed in order to investigate the correlation between the nature of mucosal inflammation as defined by both histological and cytological evaluation, and the relationship of these test results to prognosis and therapy.

Bronchopulmonary and Disseminated Granulomatous Disease Associated with Aspergillus Fumigatus and Candida Species Infection in a Golden Retriever

A seven-year-old, female golden retriever was referred for a paroxysmal, chronic cough and dyspnea, dysphagia, facial pruritus, anterior uveitis, and deteriorating general condition. A severe, mixed interstitial and alveolar pattern, with poorly defined amorphous lesions, was seen on thoracic radiographs. Multiple, whitish nodules disseminated on the hyperemic respiratory mucosa were noted on bronchoscopy. Escherichia coli and Aspergillus fumigatus were cultured from the bronchoalveolar lavage. Granulomatous lesions in numerous organs were identified during necropsy, and Aspergillus fumigatus and Candida spp. were cultured from lung and kidney tissues. Microscopic granulomatous lesions were compatible with mycotic infection; however fungal organisms were not observed.

Comparison of the value of measurement of serum galactomannan and Aspergillus-specific antibodies in the diagnosis of canine sino-nasal aspergillosis.

Serology is currently used for the diagnosis of canine sino-nasal aspergillosis (SNA). However, the accuracy of serological testing using commercially available, standardized purified antigen preparations of Aspergillus (CAPurAspAg) has only been poorly documented. The aim of the present study was to assess the diagnostic value of an agar-gel double immunodiffusion (AGDD) test and an anti-Aspergillus IgG ELISA, using CAPurAspAg and the commercially available Platelia test for the detection of serum galactomannan. Sera from 17 dogs with SNA, 18 dogs with a nasal tumour (NT), 11 dogs with lymphoplasmacytic rhinitis (LPR) and 33 control dogs were tested with the 3 methods. AGDD result was positive in 76.5% of dogs with SNA, whereas all sera from dogs with non-fungal nasal disease and control dogs were negative. A positive IgG ELISA result was obtained in 88% of dogs with SNA and in 18% of dogs with LPR. All patients with NT and control dogs had a negative IgG ELISA result. The Platelia test was positive in 24% of dogs with SNA, 11% of dogs with NT, 9% of dogs with LPR and 24% of control dogs. The results of this study suggest that (1) the detection of serum Aspergillus-specific antibodies with AGDD or ELISA, using CAPurAspAg, provides excellent specificity and good sensitivity, (2) the specificity is higher for AGDD (100%) than for ELISA (96.8%) while sensitivity is higher for ELISA (88.2%) than for AGDD (76.5%) and (3) serum galactomannan quantification with the Plateliat test is unreliable for the diagnosis of canine SNA.