Neumonias intersticiales crónicas equinas: estudio anatomopatológico e inmunocitoquímico

En este trabajo hemos realizado un estudio sobre las neumonías Intersticiales crónicas equinas estableciendo una clasificación histopatológica basada en los trabajos de Liebow (1975) y Barrios y Selman (1991) en el hombre: neumonía intersticial usual (N.I.U.=68%), neumonía bronquiolo intersticial (N.B.l.=lO%), neumonía intersticial descamativa (N.I.D.=8%), neumonía intersticial linfoide (N.l.L.=5%), neumonía intersticial bronquiolitis obliterante (N.IB .0. =5%), neumonía Intersticial eosinofílica (N.I.E.=2%) y neumonía intersticial de células gigantes (N.I.C.G.=2%), clasificándolas en función de las características lesionales y el grado de fibrosis común que presentan, Estos tipos los hemos agrupado en tres fases, las mismas que utilizamos en el estudio inmunocitoquimico. Todas estas categorías obedecen a distintas respuestas frente a diferentes agentes infecciosos, sobre todo virus (AIEqui-l) y bacterias (Streptocoeus sp., Stafi]ococos sp., Pseudomonas sp.) y presentan tres patrones macroscópicos diferentes La fase inicial se caracteriza por fenómenos exudativos-infiltrativos; la fase intermedia muestra una marcada proliferación no sólo vascular sino también celular (células fibroblásticas), y por último la fase final presenta una proliferación y acúmulo de proteínas de matriz, sobre todo tipo colágeno. Durante estas tres fases podemos observar como progresa la fibrosis pulmonar en los équidos como consecuencia del avance de la fibrosis tanto ínter como intraalveolar, dando como resultado final la imagen histológica de pulmón en panal de abeja. El estudio inmunocitoquimico reveló en la fase inicial una positividad lineal y discontlnua en el septo y lumen alveolar frente a laminina y colágeno IV, reaccionando frente a estos anticuerpos las membranas basales de los capilares y la superficie de células fibroblásticas. Esta inmunorreacción se mantiene e incluso aumenta en la fase intermedia como consecuencia del proceso de angiogénesis, quedando reducida a la membrana basal de un escaso número de vasos entre las masas fibróticas durante la fase final. La fibronectina tiene su máxima positividad en la fase inicial e intermedia presentándose no sólo como una proteína de matriz extracelular asociada a la membrana del epitelio y capilares, sino también en el citoplasma de los macrófagos activados, Ambas fases presentan una fuerte reacción inmune frente a la fibronectina especialmente en las masas perivasculares e intraalveolares con tina positividad más o menos fibrilar que es la base para la fibrinogénesis del colágeno. Sin embargo, en la fibrosis total de las neumonías intersticiales equinas se aprecia una disminución con una reacción más difusa a manera de red. Tanto el colágeno tipo 1 como el III presentaron una disminución marcada durante la primera fase, para producirse un espectacular aumento del colágeno tipo III en el intersticio y en las masas intraalveolares en la fase intermedia. El colágeno tIpo 1 presentó escasa positividad en estas primeras etapas, aunque su reacción fue muy intensa. En la fase final existe un cambio significativo en la producción de colágeno tipo III a tipo 1, pudiendo observar una positividad heterogénea en las distintas áreas de fibrosis frente a ambos tipos de colágeno. La participación de los factores de crecimiento fue muy diferente en las neumonías crónicas equinas. Así, macrófagos, células epiteliales y fibroblastos fueron identificados como fuentes del factor de crecimiento derivado de plaquetas (PDGF) durante e] proceso de fibrosis pulmonar equina, El papel que juega parece estar relacionado con un efecto mitógeno para las células epiteliales y sobre todo fibroblastos, En el caso del factor de crecimiento transformante (TGF Beta, su participación en el proceso de la fibrosis pulmonar equina, no está clara, ya que sólo actúa en las últimas fases, estimulando débilmente a los fibroblastos para incrementar la producción de colágeno, Por tanto, consideramos que el proceso de fibrosis pulmonar en las neumonías intersticiales crónicas equinas es consecuencia del estímulo mitógeno sobre los fibroblastos de la fibronectina en las primeras etapas y el PDGF en fases finales

Extracellular matrix proteins (fibronectin, collagen III, and collagen I) immunoexpression in goat tuberculous granulomas (Mycobacterium caprae)

The lesion resulting from the interaction between Mycobacterium and the host immune response is the tuberculous granuloma. Tuberculous granulomas, except in incipient stages, are partially or totally encapsulated by connective tissue. The aim of this study was to assess the immunoexpression of the extracellular matrix proteins fibronectin, collagen III, and collagen I in granulomas caused by Mycobacterium caprae in goats (Capra aegagrus hircus) to understand capsule development at diferent granuloma stages. For this purpose, a retrospective study of 56 samples of tuberculous granulomas in lung (n=30) and mediastinal lymph node (n=26) from 17 goats naturally infected with M. caprae in stages I (n=15), II (n=14) and III (n=27) was carried out. Fibronectin immunoreaction was extracellular, fibrillar-reticular in the center of stage I, II and III granulomas and peripheral in stages II and III granulomas. Collagen III immunoexpression was extracellular and fibrillar in the center of stages I, II and III tuberculous granulomas in lung and mediastinal lymph node, and progressive expression was observed in the periphery of stages II and III granulomas. Finally, collagen I immunoexpression was extracellular and fibrillar, showing a progressive loss of central expression and an increase in peripheral expression in stage III granulomas compared to stage I granulomas. Immunoexpression of these extracellular matrix proteins could help understand fibrogenesis and dating in tuberculous granuloma in both animal models and humans

Isolation and immunophenotyping by flow cytometry of canine peripheral blood and intraepithelial and lamina propria duodenal T lymphocytes

The gut associated lymphoid tissue (GALT) effector sites play a crucial role on the pathogenesis of many immunemediated gastrointestinal diseases. The lymphocytes at these effector sites are principally T cells which present important morphological, phenotypical and functional differences. Flow cytometry (FC) is one of the most commonly used techniques to characterize intestinal lymphocytes in human and animal models. Published studies with a focus on dogs for intraepithelial lymphocytes (IEL) immunophenotyping exist in very limited numbers. Moreover, no lamina propria lymphocytes (LPL) isolation protocols in the canine species have been described for FC evaluation. In addition to immune intestinal dysregulation, imbalances in the peripheral blood immune system have been described in both human and animal gastrointestinal disorders. The aim of this study was to provide a protocol for canine IEL and LPL isolation for FC immunophenotyping of T cells subsets. Specifically, T helper, T cytotoxic, activated Th and Tc lymphocytes, regulatory, double negative, double positive, IFN-γ and IL-4 producing T cells, and to compare their respective populations between these effector sites and with the blood stream compartment in healthy dogs. The potential relationship of these cells distributions with age, sex and breed was also evaluated. This study included sixteen healthy dogs of different sexes and breeds with a mean age of 4.55 ± 2.93 years old. The selected protocols for the three immune compartments showed proper cell yield, purity, viability, and the absence of phenotypic and functional disturbances. Histologically, an adequate separation of the duodenal epithelium from the lamina propria was also observed. All the proposed T cells subsets were identified in the three immune compartments studied, showing some statistically significant differences in their distributions at these locations that highlight the importance of their individual evaluation. This study provides an adequate method for canine small intestine IEL and LPL isolation for FC immunophenotyping and is key for future studies on the gastrointestinal immune system associated with different canine diseases

Coffee Silverskin Extract: Nutritional Value, Safety and Effect on Key Biological Functions

This study aimed to complete the scientific basis for the validation of a coffee silverskin extract (CSE) as a novel food ingredient according to European legislation. Nutritional value, safety, effects on biochemical biomarkers and excretion of short chain fatty acids (SCFAs) in vivo of CSE were assessed. Proteins, amino acids, fat, fatty acids, fiber, simple sugars and micronutrients were analyzed. For the first time, toxicological and physiological effects were evaluated in vivo by a repeated-dose study in healthy Wistar rats. Hormone secretion, antioxidant (enzymatic and no-enzymatic) and anti-inflammatory biomarkers, and dietary fiber fermentability of CSE (analysis of SCFAs in feces) were studied in biological samples. This unique research confirms the feasibility of CSE as a human dietary supplement with several nutrition claims: “source of proteins (16%), potassium, magnesium, calcium and vitamin C, low in fat (0.44%) and high in fiber (22%)”. This is the first report demonstrating that its oral administration (1 g/kg) for 28 days is innocuous. Hormone secretion, antioxidant or anti-inflammatory biomarkers were not affected in heathy animals. Total SCFAs derived from CSE fiber fermentation were significantly higher (p < 0.05) in male treated rats compared to male control rats. All the new information pinpoints CSE as a natural, sustainable and safe food ingredient containing fermentable fiber able to produce SCFAs with beneficial effects on gut microbiota

Prevalencia del inicio precoz de la lactancia materna

Introducción: el momento en que el recién nacido recibe la primera toma no ha sido estudiado de modo explícito y se necesitan investigaciones para evaluar las medidas de apoyo a la lactancia. Nuestro objetivo fue determinar la prevalencia del inicio precoz de la lactancia materna (IPLM) y analizar su relación con distintos factores maternos y del recién nacido. Métodos: estudio descriptivo transversal realizado durante tres años en un hospital público. La base de datos utilizada para el estudio procedió de un registro clínico electrónico. Se realizó un análisis univariado descriptivo de todas las variables y se analizó la relación existente entre el IPLM con distintos parámetros maternos y del recién nacido mediante el test de Fisher. Resultados: nuestros resultados mostraron que la prevalencia de un IPLM fue de un 88,4%, de un total de 2.683 nacimientos incluidos en el estudio. Además, se encontró asociación significativa entre este IPLM y distintos factores maternos, como la paridad (p = 0,05) y las semanas de gestación (p = 0,047), excepto con la edad (p = 0,522). Igualmente, se encontró una asociación fuerte con todos los factores del niño (p = 0,000), como el peso, el color del líquido amniótico, el test de Apgar al minuto y a los cinco minutos, el tipo de reanimación que precisaba o la necesidad de ingreso en la unidad neonatal. Conclusiones: la tasa de IPLM en nuestro ámbito de estudio es alta y está influenciada por distintos factores maternos y del recién nacido.Este estudio recibió financiación y fue apoyado por la subvención CEI2014-MP_BS_7 de CEI-BIOTIC (Ministerio de Ciencia e Innovación). Grupo de Investigación BIO277 (Junta de Andalucía). Departamento de Enfermería (Universidad de Granada)

Colistin Selection of the Mcr-1 Gene in Broiler Chicken Intestinal Microbiota

Colistin has a long story of safe use in animals for the treatment and prevention of certain bacterial diseases. Nevertheless, the first description of the mcr-1 gene showed that colistin resistance can spread by horizontal gene transfer and changed the landscape. This study aimed to assess the effect of colistin administration on the dispersion of resistance in the microbiota of day-old broiler chicks and how the presence of mcr-1 genes influences the spread of colistin resistance determinants. In this study, 100 one-day-old chicks were divided into four groups of 25 animals (G1, G2, G3, and G4). Animals from G3/G4 were challenged with mcr-1-carrying Salmonella (day 7), while colistin (600 mg/L) was administered daily to G2/G4 animals through drinking water (from day 8 to day 15). Two quantitative PCR assays were performed to compare the amount of Salmonella and mcr-1 that were present in the caecal samples. We observed that levels of mcr-1 were higher in G3/G4 animals, especially G4, due to the spread of mcr-1-carrying Salmonella. On day 21, Salmonella levels decreased in G4, reaching similar values as those for G3, but mcr-1 levels remained significantly higher, suggesting that colistin may accelerate the spreading process when mcr-1-carrying bacteria reach the gut

Desarrollo de un modelo práctico interactivo de uso docente para identificación de especies de animales mamíferos a través del pelo

Implementación de una Aplicación Interactiva en entorno Java, que permite a los estudiantes la identificación y evaluación de restos de actividad animal por la identificación a través del pelo. Los estudiantes, partiendo de una muestra biológica de pelo, pueden llegar a establecer una relación entre ésta y la especia animal a la que pertenece, mediante un proceso interactivo de preguntas y establecimiento de decisiones en cascada mediante sistema de “Árbol de Decisión

Adenovirus-vectored African Swine Fever Virus Antigens Cocktail Is Not Protective against Virulent Arm07 Isolate in Eurasian Wild Boar

African swine fever (ASF) is a viral disease of domestic and wild suids for which there is currently no vaccine or treatment available. The recent spread of ASF virus (ASFV) through Europe and Asia is causing enormous economic and animal losses. Unfortunately, the measures taken so far are insufficient and an effective vaccine against ASFV needs to be urgently developed. We hypothesized that immunization with a cocktail of thirty-five rationally selected antigens would improve the protective efficacy of subunit vaccine prototypes given that the combination of fewer immunogenic antigens (between 2 and 22) has failed to elicit protective efficacy. To this end, immunogenicity and efficacy of thirty-five adenovirus-vectored ASFV antigens were evaluated in wild boar. The treated animals were divided into different groups to test the use of BioMize adjuvant and different inoculation strategies. Forty-eight days after priming, the nine treated and two control wild boar were challenged with the virulent ASFV Arm07 isolate. All animals showed clinical signs and pathological findings consistent with ASF. This lack of protection is in line with other studies with subunit vaccine prototypes, demonstrating that there is still much room for improvement to obtain an effective subunit ASFV vaccine

Treatment with the senolytics dasatinib/quercetin reduces SARS-CoV-2-related mortality in mice

The enormous societal impact of the ongoing COVID-19 pandemic has been particularly harsh for some social groups, such as the elderly. Recently, it has been suggested that senescent cells could play a central role in pathogenesis by exacerbating the pro-inflammatory immune response against SARS-CoV-2. Therefore, the selective clearance of senescent cells by senolytic drugs may be useful as a therapy to ameliorate the symptoms of COVID-19 in some cases. Using the established COVID-19 murine model K18-hACE2, we demonstrated that a combination of the senolytics dasatinib and quercetin (D/Q) significantly reduced SARS-CoV-2-related mortality, delayed its onset, and reduced the number of other clinical symptoms. The increase in senescent markers that we detected in the lungs in response to SARS-CoV-2 may be related to the post-COVID-19 sequelae described to date. These results place senescent cells as central targets for the treatment of COVID-19, and make D/Q a new and promising therapeutic too