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

First Oral Vaccination of Eurasian Wild Boar Against African Swine Fever Virus Genotype II

African swine fever (ASF), the most significant threat to the pig industry worldwide, has spread to more than 55 countries on three continents, and it affects more than 77% of the world swine population. In the European Union, wild boar (Sus scrofa) is the most severely affected host. The main reasons for the unprecedented and constant spread of ASF in Europe are the trade activities, the continuous movement of infected-wild boar populations among regions and the lack of vaccine to prevent ASF infection. In this study, we demonstrate that oral immunization of wild boar with a non-hemadsorbing, attenuated ASF virus of genotype II isolated in Latvia in 2017 (Lv17/WB/Rie1) conferred 92% protection against challenge with a virulent ASF virus isolate (Arm07). This is, to our knowledge, the first report of a promising vaccine against ASF virus in wild boar by oral administration. Further studies should assess the safety of repeated administration and overdose, characterize long-term shedding and verify the genetic stability of the vaccine virus to confirm if Lv17/WB/Rie1 could be used for free-ranging wild boar in ASF control programs

Skin and subcutaneous mycoses in tilapia (Oreochromis niloticus) caused by fusarium oxyspoum in coinfection with Aeromonas hydrophila

Subcutaneous mycoses in freshwater fish are rare infections usually caused by oomycetes of the genus Saprolegnia and some filamentous fungi. To date, Fusarium infections in farmed fish have only been described in marine fish. Here, we report the presence of Fusarium oxysporum in subcutaneous lesions of Nile tilapia (Oreochromis niloticus). Histopathologic evaluation revealed granuloma formation with fungal structures, and the identity of the etiological agent was demonstrated by morphological and molecular analyses. Some of the animals died as a result of systemic coinfection with Aeromonashydrophila

Experimental infection with a low virulence isolate of Neospora caninum at 70 days gestation in cattle did not result in foetopathy

The Nc-Spain 1H isolate of Neospora caninum, which was newly obtained from the brain of a congenitally asymptomatic infected calf, demonstrated a reduced in vitro tachyzoite yield and viability rate, as well as low virulence in mouse models. The objective of the present study was to determine the ability of this isolate to induce foetal death in a pregnant bovine model. For this purpose, 13 naïve pregnant heifers were divided into three groups and were experimentally challenged with either 107 tachyzoites of Nc-1 (group 1, n = 5), Nc-Spain 1H (group 2, n = 5) isolates or phosphate-buffered saline (group 3, n = 3) intravenously at 70 days of gestation. After inoculation, pregnancy was monitored and dams were sacrificed when foetal death was detected. The remaining animals were slaughtered at 45 days post-infection. Maternal and foetal samples were collected for examination by histology and parasite DNA detection. Parasitaemia, specific anti-N. caninum IgG and interferon γ responses were also studied. At 3–4 weeks after infection, foetal death was detected in 3 out of 5 Nc-1-infected dams. However, no evidence of foetal death was observed in either Nc-Spain 1H-infected or control groups during the period studied. The most severe histopathological lesions were observed in the placenta and foetal organs from Nc-1-infected cattle that exhibited foetal death. It was in these animals that N. caninum DNA was more frequently detected. Parasitaemia was observed in all Nc-1-infected dams and in only 3 out of 5 Nc-Spain 1H-infected animals. The magnitude of the immune response was significantly higher in the Nc-1-inoculated group than in the group inoculated with the Nc-Spain 1H isolate. These data reveal the reduced virulence of the Nc-Spain 1H isolate in cattle

Nuevo sistema de predicción de dificultad en la ventilación a través de dispositivos supraglóticos

The aim of this study was to propose and validate a new clinical score to predict difficult ventilation through a supraglottic airway device.The authors developed a scoring system that was prospectively validated in 5532 patients. They assigned point values to each of the identified four risk factors: male, age >45 yr., short thyromental distance, and limited neck movement, their sum composing the score. The score ranged between 0 and 7 points. A score 4 or greater is associated with an increased risk of difficult ventilation through a supraglottic airway device. Our conclusion is that the new score for prediction of difficult ventilation through a supraglottic airway device is easy to use and could help anesthetists plan for difficult airway management.El objetivo de este estudio fue proponer y validar un nuevo sistema de puntuación clínica para predecir la dificultad de ventilación a través de un dispositivo supraglótico (DSG). Los autores desarrollaron un score que se validó prospectivamente en 5532 pacientes. Se asignaron valores numéricos a cada uno de los 4 factores de riesgo que se identificaron: varón, mayor de 45 años, distancia tiromentoniana (DTM) corta y movilidad cervical limitada, y su suma compuso el score. El rango de dicho score va de 0 a 7 puntos. Un score de 4 o mayor se asocia a un riesgo incrementado de dificultad de ventilación a través de DSG. Nuestra conclusión es que el nuevo score para predicción de dificultad de ventilación a través de dispositivos supraglóticos es fácil de usar y podría ayudar en el plan de manejo de la VAD por los anestesiólogos.

Human adipose tissue–derived mesenchymal stromal cells and their phagocytic capacity

This study was supported by Research Group BIO277 (Junta de Andalucia), the Department of Nursing of the University of Granada and The Cell Production Unit of the Virgen de las Nieves Hospital of Granada (Servicio Andaluz de Salud).Mesenchymal stromal cells (MSCs) have evidenced considerable therapeutic potential in numerous clinical fields, especially in tissue regeneration. The immunological characteristics of this cell population include the expression of Toll-like receptors and mannose receptors, among others. The study objective was to determine whether MSCs have phagocytic capacity against different target particles. We isolated and characterized three human adipose tissue MSC (HAT-MSC) lines from three patients and analysed their phagocytic capacity by flow cytometry, using fluorescent latex beads, and by transmission electron microscopy, using Escherichia coli, Staphylococcus aureus and Candida albicans as biological materials and latex beads as non-biological material. The results demonstrate that HAT-MSCs can phagocyte particles of different nature and size. The percentage of phagocytic cells ranged between 33.8% and 56.2% (mean of 44.37% +/- 11.253) according to the cell line, and a high phagocytic index was observed. The high phagocytic capacity observed in MSCs, which have known regenerative potential, may offer an advance in the approach to certain local and systemic infections.Junta de Andalucia BIO277Department of Nursing of the University of GranadaCell Production Unit of the Virgen de las Nieves Hospital of Granada (Servicio Andaluz de Salud

Evidence of Leishmania infantum infection in rabbits (Oryctolagus cuniculus) in a natural area in Madrid, Spain

Leishmaniasis is one of most important neglected zoonosis and remains endemic in at least 88 developing countries in the world. In addition, anthropogenic environmental changes in urban areas are leading to its emergency world-wide. Zoonotic leishmaniasis control might only be achieved by an integrated approach targeting both the human host and the animal reservoirs, which in certain sylvatic cycles are yet to be identified. Recently, hares have been pointed out as competent reservoirs of Leishmania infantum in Spain, but the role of other lagomorphs has not been clarified. Here, 69 rabbits (Oryctolagus cuniculus) from a natural area in Madrid in which a high density was present were analyzed using indirect (immunofluorescence antibody test, IFAT) and direct (PCR, culture) techniques. Fifty-seven (82.6%) of the animals were positive to at least one technique, with IFAT yielding the highest proportion of positive samples. L. infantum was isolated in 13% animals demonstrating the occurrence of infection in this setting. Our results suggest that rabbits could play a role of competent reservoir of L. infantum and demonstrate that the prevalence of infection is high in the analyzed area

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

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)