Características citológicas del lavado bronqueoalveolar y respuesta inmune humoral frente a Aspergillus fumigatus en caballos Chilotes con obstrucción recurrente de las vías aéreas

In this study, the presence and characteristics of the recurrent airway obstruction (RAO), in Chilote horses were investigated. The Chilote horse is a breed of small and very rustic horses, descendent from, the horses that were brought by the Spaniards during the colony and remained isolated in. the Chiloe island, south of Chile, for a long time. Twenty two animals, all from the same farm and living under similar conditions, were physically examined. In all cases broncheoalveolar lavage (BALF) was performed and the recovered resident cells from the respiratory tract were studied in smears stained with May Grewald - Giemsa; the level of antibodies against Aspergillus fumigatus was determined by ELISA. The results indicate that out of the total population of animals under investigation, 13 showed clinical signs compatible with RAO and 9 were normal. In all cases, the animals that were considered RAO positive showed a higher percentage of polymorphonuclear neutrophils in the BALF: furthermore, a clear positive correlation between the relative amount of neutrophilic cells present in the airways and the severity of the signs observed during the physical examination exists. All of the animals, healthy and RAO positive horses, showed high titres of antibodies against A. fumigatus, a well known allergen, commonly distributed in. the region and present in the hay/straw. Our results show that Chilote horses, even though they are a rustic bread, are very susceptible to become RAO positive with similar characteristics to those described in other breeds, moreover, all animals were sensitized to A. fumigatus antigens showing the immunogenicity and the important distribution of this fungal allergen.El objetivo de este estudio fue analizar las características citológicas del lavado bronquealveolar (LBA) y la respuesta inmune humoral en caballos Chilote con obstrucción recurrente de las vías aéreas (ORVA) contra Aspergillus fumigatus y su rol en la inducción de este cuadro alérgico. Para esto, se utilizaron 22 caballos Chilote, de los cuales 13 presentaban ORVA y 9 estaban clínicamente sanos, los cuales fueron utilizados como controles. A todos los animales se les realizó un examen físico, se les evaluó la citología pulmonar mediante un lavado bronqueoalveolar (LBA) y se les extrajo suero sanguíneo, con el propósito de determinar anticuerpos específicos contra Aspergillus fumigatus a través de un ELISA indirecto. Los resultados obtenidos de los extendidos celulares hechos a partir del LBA muestran que el porcentaje relativo de neutrófilos encontrados en los caballos con ORVA era concordante con los hallazgos obtenidos en el examen clínico. Con respecto a la prueba de ELISA se determinó que todos los animales, incluidos los controles, tenían niveles similares de anticuerpos específicos contra Aspergillus fumigatus no existiendo diferencias entre los individuos con ORVA y el grupo de animales sanos (P > 0,05). Se puede concluir que los caballos Chilote, al igual que las otras razas de caballo, tienen la predisposición de hacer ORVA al estar sensibilizados con los alérgenos inductores de este cuadro alérgico pulmonar y que existe concordancia entre el porcentaje de neutrófilos encontrados en el LBA y la gravedad de los síntomas clínicos de estos animales. Además, los antígenos de Aspergillus fumigatus tienen un rol importante en la inducción de ORVA en esta específica raza de caballo

Eosinophils in lung tissue in 2 Chilean Criollo horses with exacerbated recurrent airway obstructions "heaves"

We report on 2 cases of recurrent airway obstruction (RAO) observed in Chilean Criollo horses aged 13 and 15 years. Both animals were classified as having severe RAO. The analysis of the recovered cells of bronchoalveolar lavage fl uid (BALF) showed high percentages of eosinophils (82% and 80% in the diff erential cell counts of the 2 horses) and poor responses to bronchodilator and corticosteroid drugs. The histopathological lung evaluation showed eosinophilic infiltration in the peribronchial tissue, as well as in the lumen of the bronchi and bronchioli. In both horses, immunohistochemistry confirmed high p65 homodimer activity in bronchial cells. This report describes exacerbated heaves with eosinophilic infiltration and p65 activity in the lung tissue without any neutrophil infiltration, which is usually characteristic for this disease. © TÜBİTAK

Inhalación de esporas de Aspergillus fumigatus induce en ratones una inflamación de las vías aéreas similar a la observada en equinos con Obstrucción Recurrente de las Vías Aéreas

The aim of this study was to determine if mice exposed to Aspergillus (A.) fumigatus spores develop airway inflammation in a similar manner to Recurrent Airway Obstruction (RAO) in horses. Different groups of 2-, 5-, 6- and 8-month-old Rockefeller (RK) mice were used throughout the experiment. The mice were maintained in the animal house for 16 days and kept in cages containing hay contaminated with A. fumigatus. After 16 days of mold exposure, the mice were bled and sacrificed to obtain bronchoalveolar lavage fluid (BALF) or lung tissues for histological analysis. In addition, we measured the levels of A. fumigatus-specific antibodies by ELISA. The results demonstrated that inhalation of A. fumigatus spores in mice induced an inflammatory response in the lungs that was characterized by peribronchiolar and perivascular mononuclear infiltration and bronchiolar luminal exudates. Moreover, there was extensive folding of the airway epithelia, which is a sign of airway smooth muscle contraction and smooth muscle hypertrophy. In addition, we observed a high number of neutrophils and an increase in IgE, IgG1 and IgG2a antibodies in the BALF from mice that were older than 5 months and were exposed to A. fumigatus. These animals recovered from their respiratory distress when they were placed in a remission environment. We conclude that simple exposure of mice to moldy hay induces a condition that is similar to RAO in horses. Therefore, using this mouse model for the study of equine RAO offers the opportunity to approximate the immunologic response to airborne irritants and allergens as well as to evaluate pharmacological therapies.El objetivo de este estudio fue determinar si ratones expuestos a esporas de Aspergillus fumigatus desarrollan una inflamación alérgica de las vías respiratorias similar a la observada en equinos con Obstrucción Recurrente de las Vías Aéreas (ORVA). Para esto se utilizaron ratones Rockefeller (RK) de 2, 5, 6 y 8 meses respectivamente, los cuales se mantuvieron por 16 días en jaulas que contenían heno contaminado con A. fumigatus. Posterior a la exposición, los animales fueron sangrados y sacrificados para la obtención de suero sanguíneo, lavado broncoalveolar (LBA) y muestras de tejido pulmonar. Además, se midieron los niveles de anticuerpos específicos para A. fumigatus a través de un ELISA indirecto. Los resultados mostraron que la inhalación de esporas de A. fumigatus en ratones induce una respuesta alérgica inflamatoria en pulmones similar a lo observado en ORVA equino, con un aumento del porcentaje de neutrófilos en el LBA y cambios histológicos pulmonares característicos de una hipersensibilidad bronquial. Asimismo, se observó un aumento de los niveles de inmunoglubulinas IgE, IgG1 e IgG2a específicas para este hongo en fluidos bronquiales. Además, los animales remitieron el estrés respiratorio cuando fueron expuestos en un ambiente de remisión. Se puede concluir que la simple exposición de esporas de A. fumugatus produce una condición alérgica similar al ORVA en equinos. Por lo tanto, este modelo experimental puede ser de utilidad para el estudio de los mecanismos inmunológicos, y probar nuevas opciones terapéuticas para esta enfermedad respiratoria de los caballos

Descrição morfológica e imuno-histoquímica de um linfoma em um cordeiro de dois meses de idade usando os marcadores MUM1/IRF4, CD79a, CD3, CD10 e Ki67

This article contains a morphological description of a case of early presentation lymphoma in a lamb, characterised by its unusual clinical history and advanced lesions. The primary tumour was located in the skin in the carpal region, and metastases were observed in the brain, right eye, all the pulmonary lobes, the myocardium and endocardium of the heart, left adrenal gland, abdominal portion of the vagal nerve, serosa of the rumen, the body of lumbar vertebra L3, and the submandibular, mediastinal and mesenteric lymph nodes. Histopathological examination revealed that it was a large cell lymphoma. Immunohistochemical analysis was carried out with antibodies for MUM1/IRF4, CD79a, CD3, CD10 and Ki67, used here for the first time as markers to characterise a lymphoma in this species. The result revealed that it was a B-cell lymphomaEn este artículo se describe morfológicamente un caso de linfoma de presentación temprana en la especie ovina, el cual se caracterizó por lo inusual de su historia clínica y lo avanzado de las lesiones. El tumor primario se localizó en la piel del carpo y se observaron metástasis en el cerebro, el ojo derecho, todos los lóbulos pulmonares, el miocardio y endocardio del corazón, la glándula adrenal izquierda, el nervio vago en su porción abdominal, la serosa del rumen, el cuerpo vertebral lumbar L3, los linfonodos submandibulares, mediastínicos y mesentéricos. El examen histopatológico reveló un linfoma no Hodgkin de células grandes. El análisis inmunohistoquímico se realizó con anticuerpos para MUM-1/IRF-4, CD79a, CD3, CD10 y Ki-67, los cuales se usaron por primera vez como marcadores para caracterizar un linfoma en esta especie. El resultado reveló que se trató de un linfoma de células-B

Aspergillus fumigatus-sensitive IgE is associated with bronchial hypersensitivity in a murine model of neutrophilic airway inflammation

Neutrophils are the predominant inflammatory cells that infiltrate airways during acute exacerbation of asthma. The importance of A. fumigatus sensitization, and IgE response in the airways in patients with acute asthma is unclear. Rockefeller (RK) mice were sensitized with A. fumigatus extract protein. The animals were subsequently challenged with different degrees of A. fumigatus contamination in the cage bedding. All groups of mice were euthanized to obtain bronchoalveolar lavage fluid (BALF) for cytological and Elisa assays, and lung tissue for histological analysis. Moreover, several bioassays were conducted to determine whether BALF IgE antibodies can activate mast cells. In this study, we demonstrated that exposure of sensitized mice to a known concentration of A. fumigatus conidia produces bronchial hyperreactivity with marked neutrophilic bronchial infiltration and increased BALF IgE, capable of triggering mast cell degranulation. This study suggests that IgE may play a role in bronchial hyperreactivity associated to A. fumigatus exposure in mice. Mice sensitized and challenged with this fungus showed characteristics of severe asthma, with an increase of BALF neutrophils, histological changes consistent with severe asthma and an increase of IgE capable of triggering type I hypersensitivit

Tamoxifen induces apoptotic neutrophil efferocytosis in horses

Macrophages and neutrophils are important cellular components in the process of acute inflammation and its subsequent resolution, and evidence increasingly suggests that they play important functions during the resolution of chronic, adaptive inflammatory processes. Exacerbated neutrophil activity can be harmful to surrounding tissues; this is important in a range of diseases, including allergic asthma and chronic obstructive pulmonary disease in humans, and equine asthma (also known as recurrent airway obstruction (RAO). Tamoxifen (TX) is a non-steroidal estrogen receptor modulator with effects on cell growth and survival. Previous studies showed that TX treatment in horses with induced acute pulmonary inflammation promoted early apoptosis of blood and BALF neutrophils, reduction of BALF neutrophils, and improvement in animals’ clinical status. The aim of this study was to describe if TX induces in vitro efferocytosis of neutrophils by alveolar macrophages. Efferocytosis assay, myeloperoxidase (MPO) detection and translocation phosphatidylserine (PS) were performed on neutrophils isolated from peripheral blood samples from five healthy horses. In in vitro samples from heathy horses, TX treatment increases the phenomenon of efferocytosis of peripheral neutrophils by alveolar macrophages. Similar increases in supernatant MPO concentration and PS translocation were observed in TX-treated neutrophils, compared to control cells. In conclusion, these results confirm that tamoxifen has a direct effect on equine peripheral blood neutrophils, through stimulation of the engulfment of apoptotic neutrophils by alveolar macrophage

Eosinophils in lung tissue in 2 Chilean Criollo horses with exacerbated recurrent airway obstructions 'heaves'

We report on 2 cases of recurrent airway obstruction (RAO) observed in Chilean Criollo horses aged 13 and 15 years. Both animals were classified as having severe RAO. The analysis of the recovered cells of bronchoalveolar lavage fluid (BALF) showed high percentages of eosinophils (82% and 80% in the differential cell counts of the 2 horses) and poor responses to bronchodilator and corticosteroid drugs. The histopathological lung evaluation showed eosinophilic infiltration in the peribronchial tissue, as well as in the lumen of the bronchi and bronchioli. In both horses, immunohistochemistry confirmed high p65 homodimer activity in bronchial cells. This report describes exacerbated heaves with eosinophilic infiltration and p65 activity in the lung tissue without any neutrophil infiltration, which is usually characteristic for this disease

Concurrent Gliosarcoma and Choroid Plexus Carcinoma in a Cow

Brain tumours in cattle are uncommon and the spontaneous development of primary brain tumours of different histological types is rare in both man and animals. In man, multiple concurrent primary tumours of different types are occasionally described. We report the rare simultaneous occurrence of two different primary brain tumours, gliosarcoma and choroid plexus carcinoma, diagnosed by microscopical and immunofluorescence evaluation in an 8-year-old cow with a 2-month history of neurological disease. Gliosarcoma is a rare variant of glioblastoma multiforme, characterized by the presence of malignant filial cells and mesenchymal tissue. This tumour has not been reported previously in animals. (C) 2016 Elsevier Ltd. All rights reserved

Effect of three transport conditions on the appearance time of autolysis in samples of rainbow trout fry (Oncorhynchus mykiss)

The aim of this work was to compare three types of transport conditions on the appearance and progress of autolytic changes in samples of dead fry sent to be processed for histopathological analysis. 300 whole fry corpses were subjected to three transport conditions: submerged in water at 4 degrees C, submerged in water at 15 degrees C and dry atmosphere at 4 degrees C. Every hour from 0 to 24 hours, groups of 4 fry per condition were fixed in 10% formalin and processed using the conventional hematoxylin and eosin stain technique (H&E). The appearance time of the autolytic changes in the different segments of the kidney and liver were determined in each group, classifying the changes as early autolytic, advanced autolytic and complete autolysis. It was observed that the autolytic changes appeared first in the livers of the corpses at 4 degrees C under dry condition while last ones to be affected were the corpses submerged in water at 4 degrees C (1 and 6 hours, respectively). The first autolytic changes appeared in the kidneys of the corpses at 4 degrees C in the dry atmosphere and the last in the corpses submerged in water at 4 degrees C (3 and 9 hours, respectively). It was concluded that out of the three transport conditions studied, the one preserving for longer the histological and cytological structure of the fish tissue for histological analysis was water preservation at 4 degrees C, demonstrating the key role of the temperature and atmospheric humidity in the progress of fish autolysis

Aspergillus fumigatus-sensitive IgE is associated with bronchial hypersensitivity in a murine model of neutrophilic airway inflammation

Neutrophils are the predominant inflammatory cells that infiltrate airways during acute exacerbation of asthma. The importance of A. fumigatus sensitization, and IgE response in the airways in patients with acute asthma is unclear. Rockefeller (RK) mice were sensitized with A. fumigatus extract protein. The animals were subsequently challenged with different degrees of A. fumigatus contamination in the cage bedding. All groups of mice were euthanized to obtain bronchoalveolar lavage fluid (BALF) for cytological and Elisa assays, and lung tissue for histological analysis. Moreover, several bioassays were conducted to determine whether BALF IgE antibodies can activate mast cells. In this study, we demonstrated that exposure of sensitized mice to a known concentration of A. fumigatus conidia produces bronchial hyperreactivity with marked neutrophilic bronchial infiltration and increased BALF IgE, capable of triggering mast cell degranulation. This study suggests that IgE may play a role in bronchial hyperreactivity associated to A. fumigatus exposure in mice. Mice sensitized and challenged with this fungus showed characteristics of severe asthma, with an increase of BALF neutrophils, histological changes consistent with severe asthma and an increase of IgE capable of triggering type I hypersensitivity. (C) 2017 Elsevier Masson SAS. All rights reserved