Reactions to environmental allergens in cats with feline lower airway disease.

Objectives: Aeroallergens have been discussed as potential triggers for feline asthma (FA), which can be induced experimentally by allergen sensitization. To date, only few studies have investigated reactions to environmental allergens in cats with naturally occurring feline lower airway disease (FLAD). The aim of the study was to compare results of intradermal testing (IDT) and serum allergen-specific immunoglobulin E-(IgE) testing (SAT) in cats with FLAD, and to investigate possible associations with allergen exposure. Material and methods: Eight cats with eosinophilic airway inflammation (EI), ten cats with mixed inflammation (MI), six with neutrophilic inflammation (NI), and 24 healthy cats (HC) were included. Cats diagnosed with FLAD were assigned to the different inflammatory groups based on bronchoalveolar lavage fluid (BLAF) cytology. SAT was performed in all cats; IDT was only carried out in cats with FLAD. Information about the cats' environment and potential allergen exposure was obtained using an owner questionnaire. Results: In comparison to 83% of HC with positive reactions on SAT only 52% of cats with FLAD had positive responses (p = 0.051). Significantly more positive reactions per cat were detected on IDT than on SAT (p = 0.001). No significant difference was found for positive reactions per cat on SAT when compared between HC, NI, EI, and MI (p = 0.377). Only "slight" agreement was found for most allergens when reactions obtained in both tests in cats with FLAD were compared, except for "moderate" agreement for English plantain (k = 0.504) and Alternaria alternata (k = 0.488). Overall, no clear association between the cats' environment and allergen reactions were detected. Conclusions and clinical importance: Interpretation of allergy test results in cats with FLAD should be done in the context of clinical signs and individual factors.

Serum Allergen-Specific Immunoglobulin E in Cats with Inflammatory Bronchial Disease.

The etiology of feline inflammatory bronchial disease is poorly understood. This study compares the degree of allergen-specific serum IgE responses between cats with feline asthma, chronic bronchitis, mixed inflammation, and clinically healthy cats (HCs). The retrospective case-control study used serum from eighteen cats with eosinophilic inflammation (EI), ten with neutrophilic inflammation (NI), six with mixed inflammation (MI), and fourteen HCs. Affected cats were categorized into groups based on bronchoalveolar lavage cytology. The measurement of IgE for 34 different allergens including fungal organisms, weeds, grasses, trees, mites, and insects was performed using an indirect ELISA. Positive reactions to allergens were detected in the serum of 17/18 cats with EI, 8/10 with NI, 6/6 with MI, and 11/14 HCs (p = 0.364). When overall positive reactions were compared between groups, cats with MI (p = <0.01) had significantly more positive reactions against mite allergens than HCs. Blood eosinophils inversely correlated with the absolute amount of allergen-specific serum IgE expressed in ELISA absorbance units (EAs) (p = 0.014). Sensitization against dust mites seems to be more prevalent in cats with MI. However, positive IgE reactions can be observed in healthy and diseased cats, and, therefore, need to be interpreted in the light of clinical findings and environmental conditions of individual patients.

Cytologic, histopathologic, and immunohistochemical features of keloidal fibroma in a dog.

A five-year-old male English Bulldog was presented with a firm, well-circumscribed, 1 cm in diameter cutaneous mass on the left flank. Fine-needle aspiration (FNA) biopsy samples were collected for cytologic analysis. Cytology revealed a highly cellular sample consisting of spindle cells, numerous bundles of thick, glassy eosinophilic material (hyalinized collagen), and inflammatory cells. Spindle cells showed moderate anisocytosis and anisokaryosis, had oval nuclei with coarsely stippled chromatin, 1-3 prominent round nucleoli, and moderate amounts of wispy cytoplasm. Cells were occasionally associated with an eosinophilic extracellular matrix. Binucleated and trinucleated spindle cells were often noted. Low numbers of macrophages, small lymphocytes, and individual well-granulated mast cells were also present. The lesion was excised and submitted for histopathologic examination, revealing a well-delineated, nonencapsulated mass composed of hyalinized collagen fibers separated by spindle-shaped mesenchymal cells in the deep dermis and subcutis. Mild anisocytosis and anisokaryosis and less than one mitosis per 10 × high power fields were present. Excision of the mass was complete. The findings were consistent with a keloidal fibroma, a rare benign variant of fibroma. Neoplastic cells showed positive immunoreactivity for vimentin, and a small-to-moderate number of tumor cells showed positive immunoreactivity for α-smooth muscle actin. This is the first cytologic description of a keloidal fibroma correlated with histopathologic findings and immunolabeling. In cases where keloidal neoplasia is suspected, and since moderate cellular atypia can be present on cytologic examination even in cases of keloidal fibroma, histopathologic examination is necessary to differentiate between keloidal fibroma and keloidal fibrosarcoma.

Correlation of clinical and radiographic variables in cats with lower airway disease.

BACKGROUND: Feline lower airway disease (FLAD) is frequently associated with radiographic abnormalities. OBJECTIVES: To evaluate whether radiographic changes in cats with naturally occurring FLAD improve with treatment and if radiographic changes correlate with clinical signs. ANIMALS: Twenty-four client-owned cats newly diagnosed with FLAD, based on medical history, typical clinical signs, radiographic findings, and examination of bronchoalveolar lavage fluid, were included in the prospective study. METHODS: At 2 examination time points (days 0 and 60), an owner questionnaire, clinical examination, and thoracic radiography were carried out. Information from the questionnaire and clinical examination were evaluated on the basis of a 12-point clinical score. Radiographs were assessed using a 10-point radiographic score. Individual treatment was given to all cats over the study period, based on severity of the disease and compliance of the cat. Clinical and radiographic scores were compared statistically for both examination time points and evaluated for correlation. RESULTS: All cats showed radiographic abnormalities at initial presentation. In addition to significant improvement in clinical variables, the total radiographic score improved significantly (P = .01) during the study period, with significant improvement in the severity of bronchial (P = .01) and interstitial lung pattern (P = .04). Improvement of the clinical and radiographic score was not correlated. CONCLUSION AND CLINICAL IMPORTANCE: In addition to clinical signs, repeated radiographic examination can be used as a diagnostic tool to evaluate treatment response in cats with FLAD.

[Evaluierung der Langzeittherapie bei Katzen mit felinem Asthma und chronischer Bronchitis]. / Evaluation of long-term therapy in cats with feline asthma and chronic bronchitis.

GEGENSTAND UND ZIEL: Felines Asthma (FA) und chronische Bronchitis (CB) sind häufige entzündliche Erkrankungen der Atemwege der Katze. Obwohl beide Krankheitsbilder durch eine Infiltration mit unterschiedlichen Entzündungszelltypen gekennzeichnet sind, sind die therapeutischen Maßnahmen oft ähnlich. Über mögliche Unterschiede im therapeutischen Management dieser beiden Atemwegserkrankungen ist wenig bekannt. Ziel der Studie war es daher, bei Katzen mit FA und CB die Erst- und Langzeitbehandlung, Therapieerfolg, Nebenwirkungen und Besitzerzufriedenheit zu vergleichen. MATERIAL UND METHODEN: 35 Katzen mit FA und 11 Katzen mit CB wurden in die retrospektive Querschnittstudie eingeschlossen. Einschlusskriterien waren kompatible klinische und radiologische Befunde sowie der zytologische Nachweis einer eosinophilen Entzündung (FA) oder einer sterilen neutrophilen Entzündung (CB) in der bronchoalveolären Lavage-Flüssigkeit (BALF). Katzen mit CB wurden ausgeschlossen, wenn Hinweise auf pathologische Bakterien vorlagen. Besitzer wurden gebeten einen standardisierten Fragebogen zum therapeutischen Management und Ansprechen auf die Behandlung auszufüllen. ERGEBNISSE: Im Gruppenvergleich wurden keine statistisch signifikanten Unterschiede der Therapie festgestellt. Die meisten Katzen wurden anfänglich mit Kortikosteroiden mittels einer oralen (FA 63%/CB 64%, p=1), inhalativen (FA 34%/CB 55%, p=0,296) oder injizierbaren Applikationsform (FA 20%/CB 0%, p=0,171) behandelt. Zusätzlich wurden in einigen Fällen orale Bronchodilatatoren (FA 43%/CB 45%, p=1) und Antibiotika (FA 20%/CB 27%, p=0,682) verabreicht. In der Langzeittherapie erhielten 43% der Katzen mit FA und 36% der Katzen mit CB inhalative Kortikosteroide (p=1), orale Kortikosteroide (FA 17%/CB 36%, p=0,220) und orale Bronchodilatatoren (FA 6%/CB 27%, p=0,084) sowie phasenweise Antibiotika (FA 6%/CB 18%, p=0,238). Behandlungsbedingte Nebenwirkungen (Polyurie/Polydipsie, Pilzinfektion im Gesicht und Diabetes mellitus) wurden bei 4 Katzen mit FA und 2 Katzen mit CB registriert. Die Mehrheit der Besitzer gab an, mit dem Ansprechen auf die Behandlung äußerst oder sehr zufrieden zu sein (FA 57%/CB 64%, p=1). SCHLUSSFOLGERUNG: Signifikante Unterschiede hinsichtlich des Managements und des Therapieansprechens konnten bei beiden Erkrankungen laut Besitzerbefragung nicht festgestellt werden. KLINISCHE RELEVANZ: Laut Besitzerumfrage können chronische Bronchialerkrankungen der Katze wie Asthma und chronische Bronchitis können mit einer ähnlichen Behandlungsstrategie erfolgreich therapiert werden.

Comparison of the respiratory bacterial microbiome in cats with feline asthma and chronic bronchitis.

Objectives: While feline chronic bronchitis (CB) is known as neutrophilic bronchial inflammation (NI), feline asthma (FA) is defined as an eosinophilic airway inflammation (EI). Feline chronic bronchial disease refers to both syndromes, with similar clinical presentations and applied treatment strategies. Recent studies described alterations of the microbiota composition in cats with FA, but little is known about the comparison of the lung microbiota between different types of feline bronchial disease. The study aimed to describe the bacterial microbiota of the lower respiratory tracts of cats with FA and CB and to identify potential differences. Methods: Twenty-two client-owned cats with FA (n = 15) or CB (n = 7) confirmed via bronchoalveolar-lavage (BALF)-cytology were included. Next-generation sequencing analysis of 16S rRNA genes was performed on bacterial DNA derived from BALF samples. QIIME was used to compare microbial composition and diversity between groups. Results: Evenness and alpha-diversity-indices did not significantly differ between cats with FA and CB (Shannon p = 0.084, Chao 1 p = 0.698, observed ASVs p = 0.944). Based on a PERMANOVA analysis, no significant differences were observed in microbial composition between animals of both groups (Bray-Curtis metric, R-value 0.086, p = 0.785; unweighted UniFrac metric, R-value -0.089, p = 0.799; weighted Unifrac metric, R-value -0.072, p = 0.823). Regarding taxonomic composition, significant differences were detected for Actinobacteria on the phylum level (p = 0.026), Mycoplasma spp. (p = 0.048), and Acinetobacteria (p = 0.049) on the genus level between cats with FA and CB, with generally strong interindividual differences seen. There was a significant difference in the duration of clinical signs before diagnosis in animals dominated by Bacteriodetes (median 12 months, range 2-58 months) compared to animals dominated by Proteobacteria (median 1 month, range 1 day to 18 months; p = 0.003). Conclusions and relevance: Lung microbiota composition is very similar in cat populations with spontaneous FA and CB besides small differences in some bacterial groups. However, with disease progression, the lung microbiome of cats with both diseases appears to shift away from dominantly Proteobacteria to a pattern more dominated by Bacteriodetes. A substantial proportion of cats tested positive for Mycoplasma spp. via sequencing, while none of them tested positive using classical PCR.

Case of mastocytemia and systemic mastocytosis in a dog: ProCyte Dx and Sysmex XT-2000i scattergram findings, morphologic features, and c-kit somatic mutation analysis.

A 10-year-old female Golden Retriever was presented for a recheck after the complete removal of low-grade complex mammary carcinoma. The in-house ProCyte Dx automated counts revealed moderate regenerative anemia and moderate eosinophilia. The ProCyte Dx WBC scattergram showed a cloud in an unusual place parallel and to the right of the monocyte dot plot location. Cells were classified as either monocytes or neutrophils with no clear separation. Complete blood count analysis performed in the laboratory on a Sysmex XT-2000iV analyzer showed moderate regenerative anemia and WBC count within RI; a differential count was not provided by the instrument. On the Sysmex XT-2000iV DIFF scattergram, neutrophil and eosinophil dot plots were present at the respective locations and appeared separated, but the instrument did not provide numerical results. In addition to the normal lymphocyte dot plot location, the second cloud of cells classified as lymphocytes was displayed to the right of the monocyte dot plot area. On the WBC/BASO scattergram, the second population of cells was present above and to the right of the leukocyte cluster. Morphologic assessment of the blood smear detected mastocytemia with 16% poorly granulated and degranulated mast cells. FNAs from the liver and spleen contained large aggregates of poorly granulated mast cells. C-kit somatic mutation screening detected the presence of point mutation S479I in exon 9 of the canine c-KIT gene. This is the first description of abnormal scattergrams from ProCyte Dx and Sysmex XT-2000iV analyzers in a dog with concurrent mastocytemia and systemic mastocytosis, and where cytologic assessments of a blood smear, liver, and spleen, and c-kit somatic mutation analysis were performed.

Evaluation of barometric whole-body plethysmography for therapy monitoring in cats with feline lower airway disease.

OBJECTIVES: Feline lower airway disease (FLAD) is a common respiratory condition in cats. Traditionally, response to therapy is monitored only by evaluation of clinical signs and radiographic examination of the lungs. Barometric whole-body plethysmography (BWBP) is considered a non-invasive, well-tolerated form of measuring airway reactivity in cats. The aim of the study was to assess pulmonary function testing by BWBP for non-invasive evaluation of response to therapy in cats with FLAD and to investigate whether BWBP parameters correlate with clinical severity. MATERIAL AND METHODS: The prospective study included 25 client-owned cats, diagnosed with FLAD on the basis of their medical history, clinical signs, radiographic findings, and bronchoalveolar lavage fluid (BALF) examination. At three time points (day 0, 14, and 60), a standardised owner questionnaire, a clinical examination and BWBP measurements were carried out. Results of the questionnaire and the clinical examination were evaluated using a clinical 12-point score. Individual therapy was administered to all patients after diagnosis, based on the severity of disease and compliance of the cat. RESULTS: The total clinical score significantly improved over the entire study period (p<0.001). Significant improvement was detected for the frequency of coughing (p = 0.009), respiratory distress (p = 0.001), lung auscultation findings (p = 0.002), and general condition and appetite (p = 0.045). The BWBP parameter Penh, an indicator of bronchoconstriction, improved significantly under initial therapy between day 0 and 14 (p = 0.009). A significant correlation between Penh and the severity of auscultation findings was seen on day 0 (r = 0.40; p = 0.013). CONCLUSION: The study supports the role of Penh as a non-invasive parameter for monitoring initial treatment response in cats with FLAD. Further studies are needed to address whether other BWBP parameters might be suitable for non-invasive therapy monitoring of FLAD. Clinical evaluation is always essential in cats with FLAD to evaluate treatment response.

Cytologic features of an acanthomatous ameloblastoma in a dog.

An 8-year-old mixed breed male dog was presented with a mass on the rostral mandibular gingiva that quickly emerged 2-3 weeks prior to presentation. The mass was firm, smooth, well-circumscribed, and approximately 2 × 1 × 0.5 cm in size rostral to the left mandibular canine tooth (304). Clinical examination and radiographs were unremarkable. Cytology revealed two distinct cell populations, consisting of numerous uniform-appearing epithelial cell clusters and low numbers of individual spindle cells. Epithelial cells had mild anisocytosis and anisokaryosis, round nuclei with finely stippled chromatin, no prominent nucleoli, high N:C ratios, and low amounts of pale basophilic cytoplasm. Slender spindle cells observed had oval nuclei with no prominent nucleoli and wispy cytoplasm. On histopathologic examination, the lamina propria of the gingiva was dissected by numerous irregular and anastomosing trabeculae and islands of neoplastic epithelial cells. Neoplastic cells were focally in connection with the hyperplastic overlying epithelium. The trabeculae were surrounded and embedded by cell-rich fibrous stroma. Peripheral to the islands and trabeculae, cells were arranged in palisades, and the nuclei had an antibasilar location. The epithelial cells had prominent intercellular bridges, low amounts of cytoplasm, and one round to oval nucleus. Anisocytosis and anisokaryosis were mild to moderate, and six mitoses/10 HPF were present. Tumor cells reached the deep sample margins. Histopathologic evaluation was consistent with acanthomatous ameloblastoma. This locally aggressive neoplasm causes alveolar bone lysis and often extends beyond alveolar bone margins. Acanthomatous ameloblastoma is an important differential for rostral mandibular gingival masses containing numerous uniform epithelial cell clusters with rare slender spindle cells.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

A challenging case of lymphoproliferative disease in a cat.

A 7-year-old male castrated Maine Coon cat presented with edema of the right hindlimb and a markedly enlarged right popliteal lymph node. A CBC showed a neutropenia of 1.5 × 103 /µL. Radiographs and ultrasonographic examination were unremarkable. Cytology of the right popliteal lymph node revealed a mixed population of cells, consisting predominantly of medium to large plasmacytoid lymphocytes, low to moderate numbers of well-differentiated plasma cells and low numbers of small lymphocytes. Plasmacytoid lymphocytes had round nuclei with finely stippled chromatin and one prominent round nucleolus. Low numbers of binucleated cells and bizarre mitotic figures, and rare multinucleated cells were observed. Histopathologic examination of the lymph node showed effacement of the normal lymph node architecture by dense sheets of neoplastic cells. Round to polygonal tumor cells of intermediate size had a low to moderate amount of cytoplasm. Round to indented hyperchromatic nuclei were often eccentrically located and contained one distinct nucleolus. Anisocytosis and anisokaryosis were moderate and 21 mitoses/10 high power field (HPF) were present. Congo red staining was negative. High numbers of tumor cells were positive for lambda light chain immunoglobulin; moderate numbers stained positive for MUM-1. A clonal BCR gene rearrangement was detected with an immunoglobulin heavy chain target (IGH), immunoglobulin lambda light chain (IgL), and kappa deleting element (Kde). Differential diagnoses for the lymphoproliferative disease in this cat included lymphoplasmacytic lymphoma and myeloma-related disorder.

Toxoplasma gondii-associated cholecystitis in a cat receiving immunosuppressive treatment.

A 9-year-old female neutered domestic shorthair cat diagnosed with immune-mediated thrombocytopenia that was treated with prednisolone and cyclosporine, was presented for anorexia, vomiting, increased liver enzymes, and hyperbilirubinemia. Abdominal ultrasound revealed a markedly thickened gallbladder and common bile duct wall. Bile cytology detected severe neutrophilic inflammation and protozoal zoites. Suspected Toxoplasma gondii infection was confirmed by real-time PCR of bile. The cat was treated with clindamycin and ursodeoxycholic acid for 6 weeks, recovered and remained stable for 2 years despite ongoing immunosuppressive treatment. Thereafter, the cat was presented with suspicion of intestinal lymphoma, and recurrence of toxoplasmosis was diagnosed. Following treatment with clindamycin and prednisolone over 4 weeks the cat was euthanized. This is the first report of Toxoplasma gondii zoites detected in bile fluid from a cat with cholecystitis. Pathogenesis of toxoplasmosis in cats is still not fully understood. Although immunosuppression can represent a relevant predisposing factor, other factors, such as virulence of the parasite and genetic polymorphism of the host, can also play an important role. Toxoplasmosis should be included as a differential diagnosis in cats developing clinical signs of an inflammatory disease while receiving immunosuppressive treatment.

Use of rapid cytochemical staining to characterize fish blood granulocytes in species of special concern and determine potential for function testing.

Studies of innate immunity in fish species of special concern are essential for better understanding of their health status during hatchery rearing conditions. The cytochemical and morphological characterizations of blood granulocytes have been used to provide information about phylogenetic differences and determine the potential use of neutrophil function assays. Rapid, simple, cytochemical staining kits used routinely for staining mammalian granulocytes have been used to characterize granulocytes from blood of four fish species: Arctic grayling, cutthroat trout, June sucker, and shovelnose sturgeon. Blood smears were stained with Peroxidase 391 (myeloperoxidase, MPO), alkaline phosphatase (AP), Periodic Acid Schiff (PAS) and Diff-quick stain; examined using bright field and differential interference contrast microscopy. Granulocytes on blood smears were evaluated based on the cell morphology, and presence or absence of the specific chromogen. Presence of lymphocytes, monocytes, platelets/thrombocytes and granulocytes was determined in all fish species. Arctic grayling, June sucker, and cutthroat trout had MPO positive granulocytes, while shovelnose sturgeon heterophils had positive reaction for leukocyte AP, but not MPO. Presence of MPO indicated potential to measure oxidative burst and degranulation of neutrophil primary granules in Arctic grayling, cutthroat trout and June sucker. Absence of MPO in shovelnose sturgeon suggested use of different enzyme marker (AP) in degranulation assay for this species. Standardization of cytochemical techniques allowed for rapid screening of leukocyte types, reducing the number of fish, time and effort to select adequate neutrophil function assays to be used in studies of health status in species of special concern.