Osteopontin and fibronectin in lung tissue, serum, and bronchoalveolar lavage fluid of dogs with idiopathic pulmonary fibrosis and control dogs.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) affects West Highland white terriers (WHWTs). Osteopontin (SPP1) and fibronectin (FN1) are associated with human IPF and are overexpressed by bronchoalveolar lavage fluid (BALF) macrophages in dogs with IPF. OBJECTIVE: To investigate the value of these proteins as biomarkers of IPF. ANIMALS: West Highland white terriers (WHWTs) with IPF, control WHWTs, and terriers. METHODS: Cross-sectional observational study. Immunohistochemistry was used to localize SPP1 and FN1 in lung tissue. Serum and BALF SPP1 and FN1 concentrations were measured using canine ELISA kits and compared between groups. RESULTS: Osteopontin stained ciliated epithelial cells, smooth muscular cells, and macrophages of all included dogs, and type-II pneumocytes and extracellular matrix of all 12 diseased WHWTs, 4/6 control WHWTs, and none of the 3 terriers. Osteopontin serum concentration was higher in diseased WHWTs (n = 22; 2.15 ng/mL [0.74-5.30]) compared with control WHWTs (n = 13; 0.63 ng/mL [0.41-1.63]; P = .005) and terriers (n = 15; 0.31 ng/mL [0.19-0.51]; P < .0001), and in control WHWTs compared with terriers (P = .005). Osteopontin BALF concentrations were higher in diseased (0.27 ng/mL [0.14-0.43]) and control WHWTs (0.25 ng/mL [0.14-0.40]), compared with terriers (0.02 ng/mL [0.01-0.08]; P < .0001 and P = .003, respectively). Fibronectin (FN1) serum concentrations were lower in diseased dogs (1.03 ng/mL [0.35-1.48]) and control WHWTs (0.61 ng/mL [0.24-0.65]) compared with terriers (2.72 ng/mL [0.15-5.21]; P < .0001 and P = .0001, respectively). There was no difference in FN1 immunostaining and FN1 BALF concentrations between groups. CONCLUSIONS: Results suggest that SPP1 is involved in pathogenesis of IPF and could predispose that breed to the disease. Osteopontin serum concentration could serve as a diagnostic biomarker of IPF.

Assessment of the nasal microbiota in dogs with fungal rhinitis before and after cure and in dogs with chronic idiopathic rhinitis.

BACKGROUND: Pathogenesis of canine fungal rhinitis is still not fully understood. Treatment remains challenging, after cure turbinate destruction may be associated with persistent clinical signs and recurrence of fungal rhinitis can occur. Alterations of the nasal microbiota have been demonstrated in dogs with chronic idiopathic rhinitis and nasal neoplasia, although whether they play a role in the pathogenesis or are a consequence of the disease is still unknown. The objectives of the present study were (1) to describe nasal microbiota alterations associated with fungal rhinitis in dogs, compared with chronic idiopathic rhinitis and controls, (2) to characterize the nasal microbiota modifications associated with successful treatment of fungal rhinitis. Forty dogs diagnosed with fungal rhinitis, 14 dogs with chronic idiopathic rhinitis and 29 healthy control dogs were included. Nine of the fungal rhinitis dogs were resampled after successful treatment with enilconazole infusion. RESULTS: Only disease status contributed significantly to the variability of the microbiota. The relative abundance of the genus Moraxella was decreased in the fungal rhinitis (5.4 ± 18%) and chronic idiopathic rhinitis (4.6 ± 8.7%) groups compared to controls (51.8 ± 39.7%). Fungal rhinitis and chronic idiopathic rhinitis groups also showed an increased richness and α-diversity at species level compared with controls. Increase in unique families were associated with fungal rhinitis (Staphyloccaceae, Porphyromonadaceae, Enterobacteriaceae and Neisseriaceae) and chronic idiopathic rhinitis (Pasteurellaceae and Lactobacillaceae). In dogs with fungal rhinitis at cure, only 1 dog recovered a high relative abundance of Moraxellaceae. CONCLUSIONS: Results confirm major alterations of the nasal microbiota in dogs affected with fungal rhinitis and chronic idiopathic rhinitis, consisting mainly in a decrease of Moraxella. Besides, a specific dysbiotic profile further differentiated fungal rhinitis from chronic idiopathic rhinitis. In dogs with fungal rhinitis, whether the NM returns to its pre-infection state or progresses toward chronic idiopathic rhinitis or fungal rhinitis recurrence warrants further investigation.

Extraesophageal reflux and reflux aspiration in dogs with respiratory diseases and in healthy dogs.

BACKGROUND: Salivary bile acids are used to diagnose extraesophageal reflux (EER) and to evaluate the risk of reflux aspiration that is associated with respiratory diseases in dogs. OBJECTIVES: To study total bile acid (TBA) concentrations in saliva and in bronchoalveolar lavage fluid (BALF) to investigate EER and reflux aspiration in dogs with respiratory diseases and in healthy dogs. ANIMALS:  Thirty-one West Highland White Terriers (WHWTs) with idiopathic pulmonary fibrosis (IPF), 12 dogs with inflammatory airway disease (IAD), 6 dogs with recurrent pneumonia (RP), 26 brachycephalic dogs (BD), 27 healthy WHWTs (HW), 52 healthy dogs (HD). All privately-owned dogs. METHODS: Saliva and BALF were collected from dogs in each group. RESULTS: Salivary TBA concentrations were higher in IPF (median 0.1692 µM, interquartile range [IQR] 0.1115-0.2925 µM, Cohen's d 3.4, 95% confidence interval [CI] 2.2-4.0, P < .001) and BD (0.0256 µM, IQR 0.0086-0.0417 µM, d 0.5, CI -0.1 to 1.1, P = .003) compared to HD (0 µM, IQR not quantifiable [n.q.]-0.0131 µM). Bronchoalveolar lavage fluid TBA concentrations were higher in IPF (0.0117 µM, IQR 0.0048-0.0361 µM, d 0.5, CI 0-1.1, P < .001) compared to HD (0 µM, IQR n.q.-0.0074 µM). CONCLUSION AND CLINICAL IMPORTANCE: Extraesophageal reflux and reflux aspiration occur in healthy dogs and those with respiratory diseases.

The prevalence of pulmonary hypertension assessed using the pulmonary vein-to-right pulmonary artery ratio and its association with survival in West Highland white terriers with canine idiopathic pulmonary fibrosis.

BACKGROUND: Pulmonary hypertension (PH) is a known co-morbidity in West Highland white terriers (WHWTs) affected with canine idiopathic pulmonary fibrosis (CIPF). The pulmonary vein-to-right pulmonary artery ratio (PV/PA) has recently been described for the detection of pre-capillary PH in dogs. The objective of the present study was to estimate the prevalence of PH at diagnostic, in WHWTs affected with CIPF, by using PV/PA, in comparison with a group of healthy breed-matched controls (CTRLs). Additional study objective was to explore whether the presence of PH at initial diagnosis of CIPF impacted survival time in dogs treated with sildenafil. RESULTS: Twenty-five client-owned WHWTs presented with CIPF and 19 CTRLs were included in the study. PV/PA in either two-dimensional mode (2D) or time-motion mode or both were measured from cineloops in each dog. Dogs were classified according to PV/PA value into non/mild PH (PV/PA measured in 2D ≥ 0.7) or moderate/severe PH (PV/PA < 0.7). Survival data of WHWTs affected with CIPF were extracted from medical record to assess association between presence of PH at diagnosis and outcome. 60 % overall prevalence for moderate/severe PH was estimated in this cohort of WHWTs presented with CIPF vs. 5 % in CTRLS (P = 0.0002). The presence of moderate/severe PH at initial presentation was not associated with survival. CONCLUSIONS: Results of the present study confirm a high prevalence of PH at diagnosis in WHWTs affected with CIPF and highlight the utility of PV/PA as a non-invasive surrogate for assessment of PH in this population.

Diagnosis and treatment of gastro-oesophageal junction abnormalities in dogs with brachycephalic syndrome.

OBJECTIVES: To determine whether there is a benefit of using pre- and postoperative antacid treatment in dogs undergoing surgery for brachycephalic syndrome. To assess the use of an obstruction manoeuvre during endoscopy for the detection of dynamic gastro-oesophageal junction abnormalities. MATERIALS AND METHODS: Thirty-six client-owned brachycephalic dogs were prospectively included in a randomised trial. Antacid treatment was randomly prescribed in 18 dogs before and after surgery while the other 18 dogs did not receive any gastrointestinal medical treatment. At presentation, at the time of surgery and at recheck, digestive clinical signs and gastro-oesophageal junction abnormalities were assessed using specific scores. Gastro-oesophageal junction abnormalities were assessed during endoscopy in standard conditions as well as during endotracheal tube obstruction. This manoeuvre was also applied in an unrelated control group. RESULTS: The results suggest a beneficial effect of antacid treatment on the improvement of digestive clinical signs and lesions in dogs with brachycephalic syndrome undergoing surgery. At postsurgical control 83% of dogs had a digestive clinical score ≤1 in the treated group in contrast to 44% in the non-treated group and 39% of dogs had a gastro-oesophageal abnormalities score (during obstruction manoeuvre) ≤1 in the treated group in contrast to 16.7% in the non-treated group. The use of the obstruction manoeuvre during endoscopic assessment in a control group revealed that gastro-oesophageal junction movements are negligible in healthy animals. CLINICAL SIGNIFICANCE: The addition of antacid treatment during the pre- and postoperative period for brachycephalic dogs undergoing surgery may result in a faster and greater improvement in treated dogs. The obstruction manoeuvre is an interesting technique to improve detection of gastro-oesophageal junction abnormalities.

Assessment of the lung microbiota in dogs: influence of the type of breed, living conditions and canine idiopathic pulmonary fibrosis.

BACKGROUND: Literature about the lung microbiota (LM) in dogs is sparse. Influence of breed and living conditions on the LM in healthy dogs is currently unknown, as well as the influence of chronic respiratory diseases such as canine idiopathic pulmonary fibrosis (CIPF) in West highland white terriers (WHWTs). Aims of this study were (1) to assess the characteristics of the healthy LM according to breed and living conditions, and (2) to study LM changes associated with CIPF in WHWTs. Forty-five healthy dogs divided into 5 groups: domestic terriers (n = 10), domestic shepherds (n = 11), domestic brachycephalic dogs (n = 9), domestic WHWTs (n = 6) (H-WHWTs) and experimental beagles (n = 9) and 11 diseased WHWTs affected with CIPF (D-WHWTs) were included in the study to achieve those objectives. RESULTS: In healthy domestic dogs, except in H-WHWTs, the presence of few discriminant genera in each type of breed was the only LM modification. LM of experimental dogs displayed a change in b-diversity and an increased richness compared with domestic dogs. Moreover, Prevotella_7 and Dubosiella genera were more abundant and 19 genera were discriminant in experimental dogs. LM of both H-WHWTs and D-WHWTs revealed increased abundance of 6 genera (Brochothrix, Curvibacter, Pseudarcicella, Flavobacteriaceae genus, Rhodoluna and Limnohabitans) compared with other healthy domestic dogs. Brochothrix and Pseudarcicella were also discriminant in D-WHWTs compared with H-WHWTs and other healthy domestic dogs. CONCLUSIONS: In domestic conditions, except for H-WHWT, the breed appears to have minor influence on the LM. LM modifications were found in experimental compared with domestic living conditions. LM modifications in H-WHWTs and D-WHWTs compared with other healthy domestic dogs were similar and seemed to be linked to the breed. Whether this breed difference might be related with the high susceptibility of WHWTs for CIPF requires further studies.

Identification of Pro-Fibrotic Macrophage Populations by Single-Cell Transcriptomic Analysis in West Highland White Terriers Affected With Canine Idiopathic Pulmonary Fibrosis.

Canine idiopathic pulmonary fibrosis (CIPF) affects old dogs from the West Highland white terrier (WHWT) breed and mimics idiopathic pulmonary fibrosis (IPF) in human. The disease results from deposition of fibrotic tissue in the lung parenchyma causing respiratory failure. Recent studies in IPF using single-cell RNA sequencing (scRNA-seq) revealed the presence of profibrotic macrophage populations in the lung, which could be targeted for therapeutic purpose. In dogs, scRNA-seq was recently validated for the detection of cell populations in bronchoalveolar lavage fluid (BALF) from healthy dogs. Here we used the scRNA-seq to characterize disease-related heterogeneity within cell populations of macrophages/monocytes (Ma/Mo) in the BALF from five WHWTs affected with CIPF in comparison with three healthy WHWTs. Gene set enrichment analysis was also used to assess pro-fibrotic capacities of Ma/Mo populations. Five clusters of Ma/Mo were identified. Gene set enrichment analyses revealed the presence of pro-fibrotic monocytes in higher proportion in CIPF WHWTs than in healthy WHWTs. In addition, monocyte-derived macrophages enriched in pro-fibrotic genes in CIPF compared with healthy WHWTs were also identified. These results suggest the implication of Ma/Mo clusters in CIPF processes, although, further research is needed to understand their role in disease pathogenesis. Overexpressed molecules associated with pulmonary fibrosis processes were also identified that could be used as biomarkers and/or therapeutic targets in the future.

Idiopathic pulmonary fibrosis in West Highland white terriers: An update.

Canine idiopathic pulmonary fibrosis (CIPF) affects middle-aged to older dogs of a single breed, mainly the West Highland white terrier (WHWT), which is suggestive of a genetic predisposition. CIPF causes exercise intolerance, restrictive dyspnoea and coughing. Coarse crackles are heard on thoracic auscultation. Abnormal blood gas parameters and a shortened '6-min-walking test' distance are common; secondarily induced pulmonary hypertension and/or airway collapse are frequent. These features of CIPF mimic those of idiopathic pulmonary fibrosis (IPF) in humans and therefore identify CIPF as a possible spontaneously arising model for study of human IPF. However, computed tomographic and histopathological findings of CIPF are not identical to those of human IPF. As in human IPF, the aetiology of CIPF is not yet fully elucidated. There are no curative treatments and the prognosis is poor. This paper reviews advances in understanding of the clinical description and natural history of CIPF, the investigation of biomarkers and the exploration of possible aetiologies and mechanistic hypotheses.

Effects of manipulations to detect sliding hiatal hernia in dogs with brachycephalic airway obstructive syndrome.

OBJECTIVE: To determine the influence of manipulations aimed at increasing the transdiaphragmatic pressure gradient on the gastro-esophageal junction (GEJ) of dogs with brachycephalic airway obstructive syndrome (BAOS), and to identify the manipulation that most improves the detection of GEJ abnormalities and sliding hiatal hernia (SHH) in dogs with BAOS. STUDY DESIGN: In vivo experimental pilot study and prospective clinical study. ANIMALS: Five purpose-bred Beagles and 20 dogs diagnosed with BAOS. METHODS: Respiratory and digestive clinical signs as well as respiratory and GEJ abnormalities were scored. The presence of SHH was investigated using radiography and endoscopy in standard conditions. Endoscopic investigation was repeated after manipulations including manual pressure on the cranial abdomen (MP), Trendelenburg position (30°), or temporary complete endotracheal tube obstruction (ETO). RESULTS: No SHH was detected in any normal dog under any condition. Sixty-five percent of dogs with BAOS presented with digestive clinical signs, including vomiting and/or regurgitation. SHH was observed in only one dog via radiography and was not detected via endoscopy. Manipulations during endoscopy influenced GEJ abnormalities and allowed the detection of SHH in 2 (30°), 4 (ETO), and 5 (MP) dogs, respectively. Digestive clinical signs correlated with GEJ abnormalities observed only in dogs with ETO (P = .02). CONCLUSION: Manipulations aimed at increasing the transdiaphragmatic pressure gradient during endoscopy in BAOS dogs allowed the detection of GEJ abnormalities and SHH that were not detected under standard conditions. Although MP allowed detection of SHH in more dogs than ETO, scores under MP did not correlate with digestive clinical signs. Therefore, ETO may be more accurate manipulation for the detection of GEJ abnormalities in BAOS dogs.

The Lung Microbiome in Idiopathic Pulmonary Fibrosis: A Promising Approach for Targeted Therapies.

This review focuses on the role of the lung microbiome in idiopathic pulmonary fibrosis. Although historically considered sterile, bacterial communities have now been well documented in lungs both in healthy and pathological conditions. Studies in idiopathic pulmonary fibrosis (IPF) suggest that increased bacterial burden and/or abundance of potentially pathogenic bacteria may drive disease progression, acute exacerbations, and mortality. More recent work has highlighted the interaction between the lung microbiome and the innate immune system in IPF, strengthening the argument for the role of both host and environment interaction in disease pathogenesis. Existing published data suggesting that the lung microbiome may represent a therapeutic target, via antibiotic administration, immunization against pathogenic organisms, or treatment directed at gastroesophageal reflux. Taken altogether, published literature suggests that the lung microbiome might serve in the future as a prognostic biomarker, a therapeutic target, and/or provide an explanation for disease pathogenesis in IPF.

COMPARISON BETWEEN SEDATION AND GENERAL ANESTHESIA FOR HIGH RESOLUTION COMPUTED TOMOGRAPHIC CHARACTERIZATION OF CANINE IDIOPATHIC PULMONARY FIBROSIS IN WEST HIGHLAND WHITE TERRIERS.

Canine idiopathic pulmonary fibrosis is a progressive interstitial lung disease mainly affecting West Highland white terriers. Thoracic high-resolution computed tomographic (T-HRCT) findings for Canine idiopathic pulmonary fibrosis acquired under general anesthesia have been described previously. However, the use of general anesthesia may be contraindicated for some affected dogs. Sedation may allow improved speed and safety, but it is unknown whether sedation would yield similar results in identification and grading of Canine idiopathic pulmonary fibrosis lesions. The aim of this prospective, observational, method-comparison, case-control study was to compare findings from T-HRCT images acquired under sedation versus general anesthesia for West Highland white terriers affected with Canine idiopathic pulmonary fibrosis (n = 11) and age-matched controls (n = 9), using the glossary of terms of the Fleischner Society and a scoring system. Ground-glass opacity was identified in all affected West Highland white terriers for both sedation and general anesthesia acquisitions, although the Ground-glass opacity extent varied significantly between the two acquisitions (P < 0.001). Ground-glass opacity was the sole lesion observed in control dogs (n = 6), but was less extensive compared with affected West Highland white terriers. Identification and grading of a mosaic attenuation pattern differed significantly between acquisitions (P < 0.001). Identification of lesions such as consolidations, nodules, parenchymal and subpleural bands, bronchial wall thickening, and bronchiectasis did not differ between acquisitions. The present study demonstrated that T-HRCT obtained under sedation may provide different information than T-HRCT obtained under general anesthesia for identification and grading of some Canine idiopathic pulmonary fibrosis lesions, but not all of them. These differences should be taken into consideration when general anesthesia is contraindicated and sedation is necessary for evaluating West Highland white terriers with Canine idiopathic pulmonary fibrosis.

Assessment of CCL2 and CXCL8 chemokines in serum, bronchoalveolar lavage fluid and lung tissue samples from dogs affected with canine idiopathic pulmonary fibrosis.

Canine idiopathic pulmonary fibrosis (CIPF) is a progressive disease of the lung parenchyma that is more prevalent in dogs of the West Highland white terrier (WHWT) breed. Since the chemokines (C-C motif) ligand 2 (CCL2) and (C-X-C motif) ligand 8 (CXCL8) have been implicated in pulmonary fibrosis in humans, the aim of the present study was to investigate whether these same chemokines are involved in the pathogenesis of CIPF. CCL2 and CXCL8 concentrations were measured by ELISA in serum and bronchoalveolar lavage fluid (BALF) from healthy dogs and WHWTs affected with CIPF. Expression of the genes encoding CCL2 and CXCL8 and their respective receptors, namely (C-C motif) receptor 2 (CCR2) and (C-X-C motif) receptor 2 (CXCR2), was compared in unaffected lung tissue and biopsies from dogs affected with CIPF by quantitative PCR and localisation of CCL2 and CXCL8 proteins were determined by immunohistochemistry. Significantly greater CCL2 and CXCL8 concentrations were found in the BALF from WHWTs affected with CIPF, compared with healthy dogs. Significantly greater serum concentrations of CCL2, but not CXCL8, were found in CIPF-affected dogs compared with healthy WHWTs. No differences in relative gene expression for CCL2, CXCL8, CCR2 or CXCR2 were observed when comparing lung biopsies from control dogs and those affected with CIPF. In affected lung tissues, immunolabelling for CCL2 and CXCL8 was observed in bronchial airway epithelial cells in dogs affected with CIPF. The study findings suggest that both CCL2 and CXCL8 are involved in the pathogenesis of CIPF. Further studies are required to determine whether these chemokines might have a clinical use as biomarkers of fibrosis or as targets for therapeutic intervention.

Procollagen type III amino terminal propeptide concentrations in dogs with idiopathic pulmonary fibrosis compared with chronic bronchitis and eosinophilic bronchopneumopathy.

Idiopathic pulmonary fibrosis (IPF) is characterised by an abnormal accumulation of collagen type III in the pulmonary interstitium. Procollagen type III amino terminal propeptide (PIIINP) is used as a marker of collagen type III synthesis. In this study, the concentrations of PIIINP were investigated in dogs with IPF (n=15), dogs with chronic bronchitis (CB, n=19), dogs with eosinophilic bronchopneumopathy (EBP, n=13) and healthy dogs (n=25). PIIINP concentrations in serum and bronchoalveolar lavage fluid (BALF) were analysed by radioimmunoassay. Serum PIIINP values did not differ between groups, indicating that serum PIIINP is not useful in evaluating respiratory diseases in dogs. BALF PIIINP was significantly elevated in dogs with IPF compared with healthy dogs (P=0.002) and dogs with CB (P<0.001). BALF PIIINP was significantly higher in dogs with EBP than in dogs with CB (P=0.003) or healthy dogs (P=0.022). There were no differences in BALF PIIINP concentrations between dogs with IPF and dogs with EBP or between dogs with CB and healthy dogs. These results indicate that IPF is associated with elevated BALF PIIINP concentrations. BALF PIIINP concentrations also are elevated in EBP, possibly due to secondary fibrotic changes.

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.

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.

Treatment of 46 cats with porcine lente insulin--a prospective, multicentre study.

This prospective, multicentre, non-blinded, open study followed 46 cats with diabetes mellitus during treatment with porcine lente insulin (also known as porcine insulin zinc suspension, Caninsulin, Intervet) for 16+/-1 weeks (stabilization phase), with additional monitoring of some cats (n=23) for a variable period. At least three of the following were present at initial presentation: appropriate history of clinical signs consistent with diabetes mellitus, glucosuria, blood glucose greater than 15 mmol/l and fructosamine greater than 380 micromol/l. Insulin treatment was started at a dose rate of 0.25-0.5 IU/kg body weight twice daily, with a maximum starting dose of 2 IU/injection. Twenty-eight of the cats were classed as reaching clinical stability during the study, in 23 of these cats this was during the stabilization phase. Seven cats went into remission during the stabilization phase and one of the cats in week 56. Clinical signs of hypoglycaemia, significantly associated with a dose of 3 units or 0.5 IU/kg or more per cat (twice daily), were observed in nine of the 46 cats during the stabilization phase and concomitant biochemical hypoglycaemia was recorded in most cases. Biochemical hypoglycaemia, recorded in 6% of the blood glucose curves performed during the stabilization phase, was significantly associated with a dose rate of 0.75 IU/kg or more twice daily. This further highlights the need for cautious stepwise changes in insulin dose. The protocol used in the present study is suitable for and easy to use in practice. This study confirmed the efficacy and safety of porcine lente insulin (Caninsulin) in diabetic cats under field conditions.

Whole blood and tissue fungal DNA quantification in the diagnosis of canine sino-nasal aspergillosis.

Various combinations of tests are used to confirm the diagnosis of canine sino-nasal aspergillosis (SNA) because false-positive and false-negative results can occur with each test. Therefore, the aim of this study was to evaluate whether detection of fungal DNA in blood and nasal tissue samples was of value in the clinical diagnosis of this disease. Four groups were included in the study (dogs with SNA, lymphoplasmacytic rhinitis or nasal neoplasia, and control animals). Real-time PCR assays detecting DNA from all Penicillium and Aspergillus species (PenAsp assay) or species-specific DNA from A. fumigatus, A. terreus, A. flavus and A. niger were applied to whole blood and nasal tissue samples. Results obtained by PCR were compared between the groups. Sensitivity, specificity, positive and negative predictive values (PPV and NPV) for fungal DNA detection were compared with those for alternative diagnostic procedures including histopathology, serology and fungal culture. Significantly more fungal DNA was detected by the PenAsp assay in tissue biopsies from dogs with SNA than in the three other groups. Sensitivity, specificity, PPV and NPV for this method were 1.00, 0.06, 0.32 and 1.00. A. fumigatus DNA was detected in seven tissue biopsies from dogs with SNA and in one biopsy from a dog with a nasal tumour. Sensitivity, specificity, PPV and NPV for this diagnostic test were 0.50, 0.97, 0.87 and 0.82. No significant difference was found between the groups with respect to the amount of DNA detected in blood by the PenAsp assay. Sensitivity, specificity, PPV and NPV for this method were 0.71, 0.24, 0.31 and 0.64. A. fumigatus DNA was detected in the blood of three dogs with SNA and sixteen dogs without SNA. Sensitivity, specificity, PPV and NPV for this diagnostic tool were 0.21, 0.45, 0.15 and 0.54. Detection of A. fumigatus DNA in nasal tissue had the highest specificity, PPV and NPV but sensitivity of this method was low. Detection of fungal DNA in whole blood was of no value in the diagnosis of SNA.

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-PGF2alpha was determined. At test 1, healthy cats and cats with bronchitis were untreated, whereas for test 2 inhalant fluticasone (250 microg 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-PGF2alpha 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-PGF2alpha in BAL fluid was responsive to treatment and appeared as a sensitive biomarker of lower airway inflammation in cats.

Collection of exhaled breath condensate and analysis of hydrogen peroxide as a potential marker of lower airway inflammation in cats.

The objective of this study was to describe a standardised and non-invasive method for exhaled breath condensate (EBC) collection in cats and to test whether determination of hydrogen peroxide (H(2)O(2)) in EBC might be used as marker of lower airway inflammation. The technique of barometric whole body plethysmography for cats was combined with a system to condense the effluent air from the plethysmograph, allowing simultaneous EBC collection and respiratory pattern measurement. H(2)O(2) was determined spectrophotometrically. Eighteen experimental cats were used to investigate the impact on EBC volume and EBC H(2)O(2) of plethysmograph ventilation rate, collection duration, sample stability, within-day and day-to-day variability. After determination of a standardised EBC collection procedure, correlation analyses between EBC H(2)O(2) and bronchoalveolar lavage (BAL) cytology of healthy and allergen-challenged Ascaris suum (AS)-sensitised cats were performed. A significant and positive correlation between EBC H(2)O(2) and bronchoalveolar lavage (BAL) neutrophil% was found in healthy cats (P < 0.001, r = 0.55), whereas in AS-sensitised cats, correlation with BAL eosinophil% was significant (P < 0.005, r = 0.61). H(2)O(2) was increased after an allergen challenge in AS-sensitised cats (n = 6, 0.56+/-0.12 versus 1.08+/-0.35 micromol/L, P < 0.05). This study proposes a non-invasive, well tolerated and repeatable method of EBC collection for cats and suggests that EBC H(2)O(2) might be used as non-invasive biomarker for monitoring lower airway inflammation.