Intradermal testing and serum allergen-specific IgE-testing in cats with naturally occurring feline bronchial disease.

OBJECTIVES: While feline asthma (FA) is considered to be of allergic origin, the etiology of feline chronic bronchitis (CB) to date is unknown. Aim of the study was to compare the results of intradermal testing (IDT) and serum testing for allergen-specific immunoglobulin E (SAT) in cats diagnosed with FA and CB. MATERIAL AND METHODS: Twenty-seven client-owned cats with clinical signs, suggestive of feline inflammatory bronchial disease (FBD) were prospectively enrolled in the study. Patients were assigned to 3 groups based on results of bronchoalveolar-lavage-fluid (BALF)-cytology: FA (n=8), CB (n=10), or cats with a physiological BALF cytology (PB; n=9). A standardized IDT for 27 allergens was performed in all cats. In addition, allergen-specific IgE was measured in serum samples using an FcεRIα-ELISA. The number of positive reactions in both tests was compared between groups, and agreement between test results of both tests was evaluated. RESULTS: Regarding the number of positive reactions, no statistically significant difference was detected between groups in IDT (p=0.65) and SAT (p=0.51). When comparing the 2 test systems, a weak correlation was found for the allergens Tyrophagus putrescentiae (k=0.256), Dermatophagoides farinae (k=0.276), and rye (k=0.273). The most commonly observed reactions were to house dust mites, storage mites, rye and nettle in IDT and to sheep sorrel, storage mites, and house dust mites in SAT. CONCLUSION AND RELEVANCE: IDT and SAT in cats with feline inflammatory bronchial disease (FBD) cannot be used interchangeably for allergen detection. Sensitization to environmental allergens can occur in cats with and without airway inflammation. Therefore, a positive test result should always be assessed in context with clinical signs and allergen exposure.

Evaluation of different cleaning methods for feline inhalation chambers after bacterial contamination.

OBJECTIVES: Inhalation chambers are commonly used for the delivery of aerosol drugs to cats with respiratory disease. The aim of the study was to identify successful cleaning methods for inhalation devices after standardised bacterial contamination. METHODS: Spacer devices of two different manufacturers were used: RC Chamber (Cegla Medizintechnik) and Aerokat (Trudell Medical International). The bacterial contamination was performed using Pseudomonas aeruginosa. Previously marked areas of the chamber were contaminated with 50 µl of bacterial solution, containing between 2.2 ×105 and 2.1 ×108 colony-forming units/ml each. After cleaning the devices as recommended by each manufacturer (RC Chamber: special microwave cleaning bag [n = 5] or boiling water with liquid dish detergent for 15 mins [n = 5]; Aerokat: rinsing in a solution of lukewarm water and liquid dish detergent for 15 mins), chambers were air-dried for 24 h and samples for bacterial culture were taken from three defined areas. Sample material was applied on Müller-Hinton agar plates and subsequently incubated for 24 h at 37°C. RESULTS: Bacterial contamination was not detected in any of the examined inhalation devices using the recommended cleaning methods. CONCLUSIONS AND RELEVANCE: If inhalation chambers are cleaned following the manufacturers' recommendations, successful bacterial decontamination can be expected.

Retrospective analysis of pleural effusion in cats.

OBJECTIVES: Pleural effusion is a common presenting cause for feline patients in small animal practice. The objectives of this study were to identify possible correlations between the aetiology of effusion and clinical and laboratory findings. METHODS: In this retrospective study of 306 cats diagnosed with pleural effusion of established aetiology, cats were divided into six major groups: cardiac disease (CD), feline infectious peritonitis (FIP), neoplasia, pyothorax, chylothorax and miscellaneous. Clinical, laboratory and radiographic parameters were compared between groups. RESULTS: CD was the most common aetiology (35.3%), followed by neoplasia (30.7%), pyothorax (8.8%), FIP (8.5%), chylothorax (4.6%) and miscellaneous diseases (3.7%). In 26 (8.5%) cats, more than one underlying disease was diagnosed as a possible aetiology for pleural effusion. Cats with FIP were significantly younger than those with CD (P <0.001) and neoplasia (P <0.001). Cats with CD were presented with a significantly lower body temperature compared with cats with FIP (P = 0.022). Cats with CD had significantly higher serum alanine aminotransferase activity compared with all other cats (FIP and pyothorax, P <0.001; neoplasia and chylothorax, P = 0.02) and serum alkaline phosphatase activity compared with the pyothorax (P <0.001) and FIP groups (P = 0.04), and significantly lower protein concentrations (FIP, pyothorax and neoplasia, P <0.001; chylothorax, P = 0.04) and nucleated cell counts in the effusion than all other groups (pyothorax and neoplasia, P <0.001; chylothorax, P = 0.02; FIP, P = 0.04). The glucose level in the effusion of cats with pyothorax was significantly lower than glucose levels in patients with CD, neoplasia and chylothorax (P <0.001). Of 249 cats with a follow-up of at least 10 days, 55.8% died or were euthanased during that time. CONCLUSIONS AND RELEVANCE: CD and neoplasia were the most common causes for feline pleural effusion. Age, liver enzymes, as well as cell count, protein and glucose levels in the effusion can aid in the investigation of underlying aetiologies.

Bacterial microbiome in the nose of healthy cats and in cats with nasal disease.

BACKGROUND: Traditionally, changes in the microbial population of the nose have been assessed using conventional culture techniques. Sequencing of bacterial 16S rRNA genes demonstrated that the human nose is inhabited by a rich and diverse bacterial microbiome that cannot be detected using culture-based methods. The goal of this study was to describe the nasal microbiome of healthy cats, cats with nasal neoplasia, and cats with feline upper respiratory tract disease (FURTD). METHODOLOGY/PRINCIPAL FINDINGS: DNA was extracted from nasal swabs of healthy cats (n = 28), cats with nasal neoplasia (n = 16), and cats with FURTD (n = 15), and 16S rRNA genes were sequenced. High species richness was observed in all samples. Rarefaction analysis revealed that healthy cats living indoors had greater species richness (observed species p = 0.042) and Shannon diversity (p = 0.003) compared with healthy cats living outdoors. Higher species richness (observed species p = 0.001) and Shannon diversity (p<0.001) were found in middle-aged cats in comparison to healthy cats in different age groups. Principal coordinate analysis revealed separate clustering based on similarities in bacterial molecular phylogenetic trees of 16S rRNA genes for indoor and outdoor cats. In all groups examined, the most abundant phyla identified were Proteobacteria, Firmicutes, and Bacteroidetes. At the genus level, 375 operational taxonomic units (OTUs) were identified. In healthy cats and cats with FURTD, Moraxella spp. was the most common genus, while it was unclassified Bradyrhizobiaceae in cats with nasal neoplasia. High individual variability was observed. CONCLUSION: This study demonstrates that the nose of cats is inhabited by much more variable and diverse microbial communities than previously shown. Future research in this field might help to develop new diagnostic tools to easily identify nasal microbial changes, relate them to certain disease processes, and help clinicians in the decision process of antibiotic selection for individual patients.

Bacterial microbiome of the nose of healthy dogs and dogs with nasal disease.

The role of bacterial communities in canine nasal disease has not been studied so far using next generation sequencing methods. Sequencing of bacterial 16S rRNA genes has revealed that the canine upper respiratory tract harbors a diverse microbial community; however, changes in the composition of nasal bacterial communities in dogs with nasal disease have not been described so far. Aim of the study was to characterize the nasal microbiome of healthy dogs and compare it to that of dogs with histologically confirmed nasal neoplasia and chronic rhinitis. Nasal swabs were collected from healthy dogs (n = 23), dogs with malignant nasal neoplasia (n = 16), and dogs with chronic rhinitis (n = 8). Bacterial DNA was extracted and sequencing of the bacterial 16S rRNA gene was performed. Data were analyzed using Quantitative Insights Into Microbial Ecology (QIIME). A total of 376 Operational Taxonomic Units out of 26 bacterial phyla were detected. In healthy dogs, Moraxella spp. was the most common species, followed by Phyllobacterium spp., Cardiobacteriaceae, and Staphylococcus spp. While Moraxella spp. were significantly decreased in diseased compared to healthy dogs (p = 0.005), Pasteurellaceae were significantly increased (p = 0.001). Analysis of similarities used on the unweighted UniFrac distance metric (p = 0.027) was significantly different when nasal microbial communities of healthy dogs were compared to those of dogs with nasal disease. The study showed that the canine nasal cavity is inhabited by a highly species-rich bacterial community, and suggests significant differences between the nasal microbiome of healthy dogs and dogs with nasal disease.

Comparison of the prevalence of Mycoplasma species in dogs with and without respiratory disease.

Aim of the study was to investigate the prevalence of Mycoplasma species in dogs with and without signs of respiratory disease. Bronchoalveolarlavage fluid (BALF) and pharyngeal swabs were collected from 29 dogs with respiratory diseases (RD) and 16 dogs without signs of RD that were euthanised because of other diseases. Samples were tested for Mycoplasma species by PCR and culture, and sequencing was performed in Mycoplasma species-positive BALF samples. Pharyngeal swabs were positive for Mycoplasma species by PCR in 91.7% of dogs with RD and 86.7% of dogs without signs of RD (p = 1.000); BALF samples were PCR-positive in 37.9% of dogs with RD and 18.8% of dogs without signs of RD (p = 0.194) Mycoplasmo culture of BALF was positive in 28.6% of dogs with RD and in 18.8% without signs of RD (p = 0.730). When culture and PCR were compared, there was no significant difference in the detection rate of Mycoplasma species (p = 0.658) Sequencing detected different Mycoplasma species. Out of these, however, Mycoplasma cynos was isolated from four dogs with RD. There is no significant difference in the prevalence of Mycoplasma species between dogs with RD and dogs without evidence of RD; however, Mycoplasma cynos seems to be associated with respiratory disease.

Sampling sites for detection of feline herpesvirus-1, feline calicivirus and Chlamydia felis in cats with feline upper respiratory tract disease.

OBJECTIVES: Feline herpesvirus-1 (FHV-1), feline calicivirus (FCV) and Chlamydia felis are involved in feline upper respiratory tract disease (FURTD). Clinical signs caused by these agents can overlap, and the involvement of certain pathogens is often unpredictable. The objectives of this study were to compare detection rates of FHV-1, FCV and C felis at different sampling sites, and to investigate the correlation between positive test results and clinical signs in cats with FURTD. METHODS: Swabs were taken from the nose, pharynx, tongue and conjunctiva of 104 cats with signs of FURTD. Real-time PCR was performed on all samples for the detection of FHV-1, FCV and C felis. RESULTS: Infectious agents were identified in 93 (89.4%) cats. Of these, 55.8% were positive for FHV-1, 50.0% for FCV and 35.6% for C felis. FCV was found more frequently in the oropharynx (92.3% of FCV-positive cats) and on the tongue (90.4%) than the conjunctiva (38.5%) (P <0.001). There was no significant difference between the four sampling sites for the detection of FHV-1 and C felis. If nasal samples had also been taken, 94.9% of FHV-1-positive cats, 96.2% of FCV-positive cats and 81.1% of C felis-positive cats would have been detected. CONCLUSIONS AND RELEVANCE: The oropharynx can be recommended as the preferred single sampling site for the detection of FCV, FHV-1 and C felis if only one sample can be taken; however, taking samples at different sites significantly increases the detection rate for all pathogens studied. Interestingly, sampling from a site with FURTD-associated lesions did not increase the likelihood of detecting the infectious agents.

Detection of feline Mycoplasma species in cats with feline asthma and chronic bronchitis.

Little is known about the aetiology of inflammatory lower airway disease in cats. The aim of this study was to investigate the role of Mycoplasma species in cats with feline asthma (FA) and chronic bronchitis (CB). The study population consisted of 17 cats with FA/CB, and 14 sick cats without clinical and historical signs of respiratory disease, which were euthanased for various other reasons. Nasal swabs, nasal lavage and bronchoalveolar lavage fluid (BALF) samples were taken from patients from both groups. Mycoplasma species culture with modified Hayflick agar and Mycoplasma polymerase chain reaction (PCR) were performed on all samples followed by sequencing of all Mycoplasma species-positive samples for differentiation of subspecies. PCR testing detected significantly more Mycoplasma species-positive BALF samples than Mycoplasma culture (P = 0.021). When cats with oropharyngeal contamination were excluded from comparison, the numbers of Mycoplasma species-positive BALF samples in the group with FA/CB (6/17) and the control group (4/9) were not significantly different (P = 0.6924). While all nasal samples of the cats with FA/CB were negative for Mycoplasma organisms, five samples in the control group (P = 0.041) were positive on PCR. Sequencing revealed Mycoplasma felis in all PCR-positive samples. Mycoplasma species can be detected in the lower airways of cats with FA/CB, as well as in the BALF of sick cats without respiratory signs. Further studies are warranted to investigate the possibility that Mycoplasma species represent commensals of the lower respiratory tract of cats.

Prevalence of Angiostrongylus vasorum and Crenosomo vulpis in dogs in Bavaria.

Worldwide there is increasing awareness of canine lung worm infection in many geographic areas. Especially the metastrongyloid nematode Angiostrongylus (A.) vasorum is associated with a high morbidity and mortality rate in infected dogs. Other pulmonary parasites in dogs include the common fox parasite Crenosoma (C.) vulpis, and the pulmonary parasites Filaroides (Oslerus) osleri, Filaroides hirthi, and the capillarid lungworms Eucoleus aerophilus (Capillaria aerophila) and Eucoleus boehmi. Aim of the study was to investigate the prevalence of A. vasorum and C. vulpis in dogs in Bavaria, Germany, and to evaluate clinical, laboratory, and radiographic changes of dogs with these pulmonary parasites. In total, three separate faecal samples from three consecutive days were collected from 276 dogs from Bavaria and analyzed by Baermann faecal technique. Included were 160 sick dogs that showed acute or chronic respiratory problems (n = 90), neurological signs (n = 47), or coagulopathies (n = 23), as well as 116 healthy controls. A. vasorum was detected in one patient with respiratory signs, and C. vulpis was found in another patient with respiratory problems. All dogs exhibiting neurological signs or coagulopathies and all healthy dogs tested negative. Thus, prevalence for both A. vasorum and C. vulpis infection was 0.36% in all dogs tested, 0.63% in all sick dogs, and 1.11% in all dogs with respiratory signs. The study shows that in contrast to the situation in other parts of Germany and in endemic areas in other European countries, the risk of infection with pulmonary parasites is low for dogs in Bavaria at the moment.

Two outbreaks of virulent systemic feline calicivirus infection in cats in Germany.

Over the last years, several outbreaks of virulent systemic feline calicivirus (VS-FCV) infection have been described in the USA and several European countries. The paper describes two outbreaks of VS-FCV infection in cats in Germany. Data concerning clinical, laboratory, and histopathological features ofVS-FCV infection were collected from two outbreaks affecting 55 and 4 cats, respectively. Presence of feline calicivirus was confirmed by PCR followed by sequencing of the PCR-products. Clinical signs were variable, including severe upper respiratory tract infection, dyspnoea, oral and footpad ulceration, facial oedema, enteritis, pneumonia, bleeding disorder, high fever, and icterus. Both outbreaks were characterized by a high mortality rate.The present report describes the first documented outbreaks of VS-FCV infection in cats in Germany. Clinical and histopathological features are comparable to outbreaks described in the USA and Europe. However, phylogenetic analysis of the virus genome suggests that virus strains involved in these outbreaks were different from each other and from virulent strains isolated before, confirming the known genetic variability of FCV.

"Candidatus Neoehrlichia mikurensis" infection in a dog from Germany.

"Candidatus Neoehrlichia mikurensis" is a new intracellular pathogen associated with human infection and death. "Candidatus Neoehrlichia mikurensis" infection in a chronically neutropenic dog from Germany was confirmed by DNA sequencing. The same organism was previously described from ticks and two sick human beings from Germany.

Use of propentofylline in feline bronchial disease: prospective, randomized, positive-controlled study.

Propentofylline is a methylxanthine derivative with bronchodilating actions similar to those of theophylline. Nineteen cats with bronchial disease were enrolled in this study. All cats received a low dose of prednisolone; 10 of the cats additionally received propentofylline. Propentofylline-treated cats significantly improved in their auscultation scores, respiratory pattern scores, and radiological bronchial markings score over the observation period, and they coughed less and slept less at the end of the study. No significant changes were noted in the control group. This study provides evidence that a combination therapy with prednisolone and propentofylline in cats with bronchial disease might be superior over monotherapy with prednisolone.