El objetivo fundamental del grupo se basa en el estudio de enfermedades de alto impacto, ...el sector de la producción animal, mediante el análisis de sus manifestaciones morfológicas a través de la necropsia y el estudio histopatológico. Integramos nuestros resultados con diversas disciplinas (biología, clínica, microbiología, inmunología, parasitología, genética, farmacología, diagnóstico por imagen...), logrando un abordaje multidisciplinar de las enfermedades para el desarrollo de biomarcadores y técnicas de diagnóstico, o el diseño de medidas de prevención, control y tratamiento. Nuestro campo de aplicación se extiende a especies animales domésticas y salvajes, animales de experimentación y animales acuáticos, habiéndonos especializado en los últimos 20 años en el campo de la ictiopatología. La experiencia adquirida en salud de los peces y experimentación animal ha llevado al grupo a desarrollar una nueva línea de investigación en el ámbito de la biomedicina empleando modelos animales, principalmente, pez cebra.
MARIBEL QUIROGA BERDEAL 982822304
Líneas de Investigación
Desarrollo de modelos experimentales de enfermedades animales y humanas en pez cebra
La experiencia adquirida en salud de los peces y experimentación animal ha llevado al grupo a desarrollar una nueva línea de investigación empleando el pez cebra como modelo para el estudio de enfermedadades de interés en salud pública y medicina veterinaria y humana.
Evaluación de la esqueletogénesis, anomalías esqueléticas y musculares en diferentes fases de producción de peces de cultivo.
el objetivo de esta línea de investigación es el abordaje multidisciplinar de las principales anomalías esqueléticas, el crecimiento y el desarrollo muscular en especies de interés en acuicultura, principalmente el lenguado senegalés, mediante el empleo de técnicas radiológicas, morfopatológicas y moleculares, con el propósito de desarrollar herramientas encaminadas al diagnóstico y monitorización de las anomalías esqueléticas y el crecimiento en diferentes condiciones de producción.
Estudio morfopatológico y de respuesta inmunitaria local en mucosas
El objetivo de esta línea de investigación es abordar el conocimiento de las mucosas intestinal, branquial y cutánea, con el propósito de mejorar la salud y desarrollar protocolos de evaluación del efecto de diatas y tratamientos. Para ello realizamos un enfoque multidisciplinar mediante el empleo de técnicas histopatológicas, inmunitarias y genéticas.
Patogenia y diagnóstico de enfermedades comunes y emergentes en mamíferos, aves y animales acuáticos, con especial énfasis en especies de intéres en acuicultura
Caracterización morfopatológica y patogénica de enfermedades comunes y emergentes, sobre todo, en aquellas que tienen un impacto enla producción animal y el medio ambiente, mediante el empleo de técnicas histopatológica,s inmunohitoquímicas, moleculares y de microscopía electrónica. El propósito final es la obtención de nuevas herramientas de diagnóstico y biomarcadores que sean de aplicación práctica al diagnóstico, prenvención, control de enfermedades y monitorización de l salud y el binestar animal,
Proyectos de Investigación
Control de la enteromixosis del rodaballo y espáridos. Caracterización de la patogenia, la interacción parásito-hospedador y la respuesta inmunitaria.
Año: 2009 Convocatoria: Ministerio de Ciencia e Innovación (AGL2009-13282-C02-2)
Conservación ex situ de la náyade en peligro de extinción Margaritifera margritifera: Implementación de la actuación de cultivo del proyecto Life+ Margal Ulla (CAQ2016)
Año: 2017 Convocatoria: Fundación Biodiversidad. Ministerio de Agricultura y Pesca y Alimantación y Medio Ambiente
Año: 2019 Convocatoria: Consellería de Educación, Universidade e Formación Profesional. Plan Galego
Búsqueda de herramientas de diagnóstico precoz y detección de biomarcadores de salud/enfermedad intestinal en rodaballo.
Año: 2015 Convocatoria: Ministerio de Economía, Industria y Competitividad. RETOS 2015 (2015-PN267)
Servicios de Inv. transferibles / Instalaciones
Servicio de diagnóstico anatomopatológico veterinario
• Prestación de servicios a la administración pública, empresas de producción animal, profesionales veterinarios, zoos y acuarios para el diagnóstico anatomopatológico de enfermedades en diferentes especies animales.
• Prestación de servicios y colaboración científica con empresas y centros/grupos de investigación para la evaluación de los efectos de dietas / tratamientos / vacunas en especies animales, así como para la optimización de protocolos de producción.
• Asesoramiento a empresas y centros/grupos de investigación en la evaluación del estatus sanitario y bienestar en animales de producción y experimentación (los miembros del equipo disponen de los certificados de capacitación en materia de protección de animales de experimentación).
Skeletal anomalies in reared Senegalese sole (Solea senegalensis) juveniles: a radiographic approach.
Reared Senegalese sole Solea senegalensis Kaup show a high incidence of vertebral anomalies; however, little is known about its skeletal anomaly profile in the later farming phases. The purpose of this study was to provide a detailed description and quantification of the most common skeletal anomalies in reared Senegalese sole in the juvenile stage by means of computed radiography. A total of 374 Senegalese sole were classified according to the external morphology of the fish as normal or altered and then radiographed in latero-lateral and in dorso-ventral projections. Radiographic evaluation of anomalies focused especially on vertebral body anomalies (VBA) and vertebral column deviations (VCD). The 2 orthogonal projections provided a more complete visualization of the skeleton. Approximately 75% of the individuals showed at least 1 anomaly, while VBA and/or VCD were detected in 48.9% of the specimens. Regarding external morphology, 88% of the fish were categorized as normal, although about 72% of these normal fish displayed abnormalities in radiographies. The most frequent anomalies consisted of deformations of the caudal complex plates (hypurals, parhypural and epural), preurals and caudal vertebrae. Scoliosis was the most prevalent among VCD, affecting the caudal area in almost 15% of the individuals. The anomaly profile at the juvenile stages showed some differences compared to what has been reported previously in earlier stages of development. In light of these results, further investigation into the progression of skeletal anomalies over time and the causative factors at later stages is required.
First description of a natural infection with spleen and kidney necrosis virus in zebrafish.
Zebrafish has become a popular research model in the last years, and several diseases affecting zebrafish research facilities have been reported. However, only one case of naturally occurring viral infections was described for this species. In 2015, infectious spleen and kidney necrosis virus (ISKNV ) was detected in zebrafish from a research facility in Spain. Affected fish showed lethargy, loss of appetite, abnormal swimming, distention of the coelomic cavity and, in the most severe cases, respiratory distress, pale gills and petechial haemorrhages at the base of fins. Cytomegaly was the most relevant histopathological finding in organs and tissues, sometimes associated to degenerative and necrotic changes. ISKNV belongs to the relatively newly defined genus Megalocytivirus , family Iridoviridae, comprising large, icosahedral cytoplasmic DNA viruses. This is the first case of naturally occurring Megalocytivirus infection in zebrafish research facilities, associated with morbidity. The virus has been identified based on both pathologic and genetic evidence, to better understand the pathogenesis of the infection in zebrafish and the phylogenetic relationship with other iridoviruses. Given the ability of megalocytiviruses to cross‐species boundaries, it seems necessary to implement stringent biosecurity practices as these infections may invalidate experimental data and have major impact on laboratory and cultured fish.https://onlinelibrary.wiley.com/doi/abs/10.1111/jfd.12822
Susceptibility of Zebrafish to Vesicular Stomatitis Virus Infection.
The zebrafish, Danio rerio, has become recognized as a valuable model for infectious diseases. Here we evaluated the susceptibility of zebrafish to be infected with the mammalian vesicular stomatitis virus (VSV). Both zebrafish cells and embryos were highly susceptible to VSV infection. Mortalities exceeded 80% in infected embryos and were preceded by the invasion of the central nervous system by VSV. Live imaging of the infection with GFP-VSV as well as virus titration from infected fish confirmed the viral replication. Immunohistochemical analysis of embryonic fish provided evidence of viral antigens as well as of the apoptosis marker caspase-3 in the brain, eye, liver, pronephros, and skeletal muscle. So far, this is the first report describing the susceptibility of zebrafish to the mammalian virus VSV.https://doi.org/10.1089/zeb.2017.1499
Long-term affected flat oyster (Ostrea edulis) haemocytes show differential gene expression profiles from naïve oysters in response to Bonamia ostreae
European flat oyster (Ostrea edulis) production has suffered a severe decline due to bonamiosis. The responsible parasite enters in oyster haemocytes, causing an acute inflammatory response frequently leading to death. We used an immune-enriched oligo-microarray to understand the haemocyte response to Bonamia ostreae by comparing expression profiles between naïve (NS) and long-term affected (AS) populations along a time series (1 d, 30 d, 90 d). AS showed a much higher response just after challenge, which might be indicative of selection for resistance. No regulated genes were detected at 30 d in both populations while a notable reactivation was observed at 90 d, suggesting parasite latency during infection. Genes related to extracellular matrix and protease inhibitors, up-regulated in AS, and those related to histones, down-regulated in NS, might play an important role along the infection. Twenty-four candidate genes related to resistance should be further validated for selection programs aimed to control bonamiosis.https://doi.org/10.1016/j.ygeno.2018.04.002
Skeletal Anomalies in Senegalese Sole (Solea senegalensis), an Anosteocytic Boned Flatfish Species.
Skeletal anomalies affect animal welfare and cause important economic problems in aquaculture. Despite the high frequency of skeletal problems in reared Solea senegalensis, there is lack of information regarding the histological features of normal and deformed vertebrae in this flatfish. The aim of this study was to describe the histopathological and radiographical appearance of vertebral body anomalies. Sixty-seven juvenile fish were radiographically examined 104 or 105 days after hatching. Through radiographic images, vertebral segments were selected and processed for histopathological examination from 7 normal and 7 affected fish. Alterations in bone shape and vertebral fusion were the most significant anomalies in the vertebral bodies. These alterations occurred most frequently between the last 3 abdominal vertebrae and the first 10 caudal centra. Radiographically, deformed vertebrae showed flattening of the endplates and narrowing of the intervertebral spaces. The radiographic findings concurred with the histological lesions where affected vertebrae exhibited irregular endplates and changes in trabecular bone. Radiolucent cartilaginous tissue was evident in the endplates of the deformed vertebra and, in some cases, the cartilaginous material extended from the growth zone into the intervertebral space. These changes were likely the primary alterations that led to vertebral fusion. Fused vertebrae were often reshaped and showed a reorganization of the trabeculae. The formation of metaplastic cartilage is frequent in a variety of anomalies affecting teleost species.https://doi.org/10.1177/0300985818800027
Integrating genomic and morphological approaches in fish pathology research: The case of turbot (Scophthalmus maximus) enteromyxosis.
Enteromyxosis, caused by Enteromyxum scophthalmi, is one of the most devastating diseases stemming from myxozoan parasites in turbot (Scophthalmus maximus L.), being a limiting factor for its production. The disease develops as a cachectic syndrome, associated to catarrhal enteritis and leukocytic depletion, with morbidity and mortality rates usually reaching 100%. To date, no effective treatment exists and there are different unknown issues concerning its pathogenesis. The gross and microscopic lesions associated to enteromyxosis have been thoroughly described, and several morphopathological studies have been carried out to elucidate the mechanisms of this host-parasite interaction. More recently, efforts have been focused on a multidisciplinary approach, combining histopathology and transcriptome analysis, which has provided significant advances in the understanding of the pathogenesis of this parasitosis. RNA-Seq technology was applied at early and advanced stages of the disease on fishes histologically evaluated and classified based on their lesional degree. In the same way, the transcriptomic data were analyzed in relation to the morphopathological picture and the course of the disease. In this paper, a comprehensive review of turbot enteromyxosis is presented, starting from the disease description up to the most novel information extracted by an integrated approach on the infection mechanisms and host response. Further, we discuss ongoing strategies toward a full understanding of host-pathogen interaction and the identification of suitable biomarkers for early diagnosis and disease management strategies.https://doi.org/10.3389/fgene.2019.00026
Early stages of Margaritifera margaritifera glochidiosis in Atlantic salmon: Morphopathological characterization.
Freshwater mussels of the order Unionida encyst into the fish mucosa to metamorphose and complete their life cycle, causing a parasitic disease known as glochidiosis. This parasitic stage represents a bottleneck for the survival of naiads, particularly for critically endangered species as Margaritifera margaritifera; however, little is known about the events occurring during this critical stage. Therefore, this study aimed to histologically characterize the development of M. margaritifera glochidiosis in Atlantic salmon to get insight into the pathogenesis of this interaction. Fish exposed to glochidia were sampled during the first 44 days post‐exposure, and organs were observed by stereomicroscopy and light microscopy. Glochidia attached to the gills by pinching the lamellar epithelium, whereupon an acute proliferative branchitis engulfed most of the larvae. However, during the first 14 days, a severe detachment of unviable glochidia occurred, associated with the presence of pleomorphic inflammatory infiltrate and epithelial degeneration. In the cases where larvae remained attached, a chronification of the lesions with none to scarce inflammation was observed. These results provide key information to better understand the complex host–parasite interaction during the early stages of glochidiosis and provide valuable information to optimize artificial rearing of naiads in conservation of threatened freshwater mussel populations.https://doi.org/10.1111/jfd.13100
Immunohistochemical expression of E–cadherin in different tissues of the teleost fish Scophthalmus maximus.
E–cadherin is an evolutionary conserved protein, whose main role as the principal component of adherens junctions is supporting epithelial cell–cell adhesion. It is an essential molecule for the maintenance of the epithelial barrier function and the analysis of its immunohistochemical expression is a valuable resource in morphopathological, ontogenetic and pathogenesis studies in mammals. As well, there is an increasing understanding of the importance of E-cadherin in the physiology of the immune system and the development of the immune response. Mucosal health is a primary issue in aquaculture research; nevertheless, there is a lack of immunohistochemical studies of cell junction proteins in fish species. In this work, an immunohistochemical technique was optimized in Bouin- and formalin-fixed paraffin-embedded tissues of turbot Scophthalmus maximus, employing a commercial antibody raised against human E-cadherin. The specificity of the antibody in recognizing the molecule in this teleost species was tested by western blot and mass spectrometry-based proteomic analyses. The assays showed a good specificity and indicated that the antibody recognizes the well conserved cytoplasmic domain of the protein. Immunohistochemistry showed the localisation of E-cadherin at cell-cell contact in the epithelia of the different organs, between the hepatocytes and the pancreatic acinar cells, as well as in the reticulo-epithelial stroma of the thymus. Also, the immunoreaction was observed in the cells constituting the melano-macrophage centres in the spleen and kidney. No immunostaining was detected, as expected, only in the heart and brain. No significant difference was noticed between the two fixative used for collecting the tissues samples. This is the first description of E-cadherin immunohistochemical expression in several tissues of a teleost. The immunohistochemical technique represents a useful tool to be used in the different areas of fish health research.https://doi.org/10.1016/j.aquaculture.2018.12.009
Effects of Enteromyxum spp. (Myxozoa) infection in the regulation of intestinal E-cadherin: turbot against gilthead sea bream
ABSTRACT:Enteromyxoses are relevant diseases for turbot and gilthead sea bream aquaculture. The myxozoan parasites invade the intestinal mucosa, causing a cachectic syndrome associated with intestinal barrier alteration; nonetheless, their pathological impact is different. Turbot infected by Enteromyxum scophthalmi develop more severe intestinal lesions, reaching mortality rates of 100%, whereas in E. leei ‐infected gilthead sea bream, the disease progresses slowly, and mortality rates are lower. The mechanisms underlying the different pathogenesis are still unclear. We studied the distribution and expression changes of E‐cadherin, a highly conserved protein of the adherens junctions, in the intestine of both species by immunohistochemistry and quantitative PCR, using the same immunohistochemical protocol and common primers. The regular immunostaining pattern observed in control fish turned into markedly irregular in parasitized turbot, showing an intense immunoreaction at the host–parasite interface. Nevertheless, E‐cadherin gene expression was not significantly modulated in this species. On the contrary, no evident changes in the protein distribution were noticed in gilthead sea bream, whereas a significant gene downregulation occurred in advanced infection. The results contribute to the understanding of the different host–parasite interactions in enteromyxoses. Host and parasite cells appear to establish diverse relationships in these species, which could underlie the different pathological picture.https://doi.org/10.1111/jfd.13130