Person: Díaz-Regañón Fernández, David Rafael
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First Name
David Rafael
Last Name
Díaz-Regañón Fernández
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Veterinaria
Department
Medicina y Cirugía Animal
Area
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14 results
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Item Prevalence and appropriateness of omeprazole prescription in dogs at a veterinary teaching hospital before and after the publication of the ACVIM consensus statement on the rational administration of gastrointestinal protectants(Frontiers in Veterinary Science, 2024) Sainz Rodríguez, Ángel; García-Sancho Téllez, Mercedes Guadalupe; Villaescusa Fernández, Alejandra; Rodríguez Franco, Fernando; Díaz-Regañón Fernández, David Rafael; Olmeda García, Patricia; Marks, Stanley L.Introduction: Overprescribing of acid suppressants is a common phenomenon in human and small animal patients, leading to potential deleterious gastrointestinal (GI) and non-GI consequences. The impact of consensus statements on veterinary prescribing habits in clinical practice have not been fully evaluated. This study aimed to compare the prescribing habits of the proton pump inhibitor (PPI), omeprazole, in dogs in an academic veterinary teaching hospital before and after the publication of the American College of Veterinary Internal Medicine (ACVIM) consensus statement on rational use of gastrointestinal protectants. Methods: Evaluation of the prescribing habits of omeprazole in dogs during the years 2017 and 2021 was retrospectively compared. These years were selected to reflect a 12-month period prior to and following the publication of the consensus statement. One hundred dogs from each year were randomly selected. Dose, frequency of administration, duration of treatment, concurrent prescription of more than one gastroprotectant and indications for prescribing omeprazole were analyzed. Results: A significant increase in the cases that received omeprazole q12h (p < 0.0001) or that underwent a tapering dose after ≥4 week-therapy (p > 0.0001) was detected after the publication of the 2018 ACVIM consensus statement. Considering the indications, there was also a significant increase in the appropriate prescription of omeprazole in the second compared to the first period of study (p < 0.0001). Fifteen of 16 clinicians (94%) involved in the prescription of omeprazole indicated that their reading of the consensus statement had changed their clinical practice regarding PPI administration in dogs. Discussion: These results support the beneficial impact of the ACVIM consensus statement on the judicious prescribing of omeprazole in an academic veterinary hospital. These results should not be extrapolated to first-opinion veterinary practices, and further efforts should be made to ensure that PPIs are prescribed prudently with a clear indication and regular review of the appropriateness of continued administration to minimize possible risks and adverse drug interactions.Item Isolation and immunophenotyping by flow cytometry of canine peripheral blood and intraepithelial and lamina propria duodenal T lymphocytes(Veterinary Immunology and Immunopathology, 2021) Agulla Pérez, Beatriz; García-Sancho Téllez, Mercedes Guadalupe; Sainz Rodríguez, Ángel; Rodríguez Franco, Fernando; Díaz-Regañón Fernández, David Rafael; Rodríguez Bertos, Antonio Manuel; Villaescusa Fernández, AlejandraThe 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.Item Assessing the Quality of Life of Dogs with Inflammatory Bowel Disease and Their Owners(Veterinary Sciences, 2023) Díaz-Regañón Fernández, David Rafael; Sainz Rodríguez, Ángel; Rodríguez Franco, Fernando; Villaescusa Fernández, Alejandra; Olmeda, Patricia; Morcillo, Ana; García-Sancho Téllez, Mercedes GuadalupeThe aim of the study was to assess the quality of life (QoL) of dogs with inflammatory bowel disease (IBD) and the impact on the life and relationship of the owner. An online questionnaire based on a Likert scale score (1–10) was designed to assess items related to dog QoL, owner QoL, and the owner–dog relationship. Responses from 110 dog owners (30 with IBD and 80 healthy dogs) were included in the study. IBD dogs had significantly lower overall QoL (p < 0.001), health (p < 0.0001), and level of activity (p = 0.049). Owners of dogs with IBD reported lower overall QoL (p < 0.001). The scores for how their dog’s QoL might affect their own QoL (p = 0.028), how much their dog limited their social life, leisure time, or daily activities (p = 0.015), and how often they felt burdened by caring for their dog (p = 0.006) were significantly higher in the IBD group when compared to the healthy group. In addition, IBD dog owners were more likely to see their dogs as children (p = 0.0004). IBD has a negative impact on affected dogs and their owners regardless of the severity of the diseasItem Animal Research in Spain: A Study of Public Perception and Attitudes(Animals, 2023) Miguel-Batuecas, Andrea; Fuertes Recuero, Manuel; Díaz-Regañón Fernández, David Rafael; Ortiz Díez, Gustavo; De Pablo Moreno, Juan AndrésSince the time of Hippocrates in the 4th century BC, animal research has been extensively used for various purposes up to the present day. However, the use of animals for research has also been controversial for a long time. We report the findings of a public, online questionnaire-based survey designed to assess the opinions of a sample of Spanish society regarding animal research. Demographic data and opinions were obtained from 806 respondents. The results indicated a high level of acceptance of animal research (73.1%). However, certain factors, such as completing the questionnaire immediately after a reading negative media report (OR = 2.41; 95%CI: 1.64–3.54; p < 0.001), being a woman (OR = 1.77; 95%CI: 1.24–2.53; p = 0.002) or having a non-scientific background (OR = 2.47; 95%CI: 1.76–3.47; p < 0.001), were associated with a tendency towards a more negative opinion. The opinions seemed to be influenced by gender, education level and by protest incidents reported in the media. Our results also indicate that a lot of information regarding animal welfare, such as care and handling protocols, along with legislation was unknown to individuals. Further, a growing popularity of companion species and opposition to animal experimentation for non-biomedical purposes were reflected in the responses obtained. The use of animals for research purposes emerged as a sensitive social issue in terms of concerns about animal ethics and welfare.Item Stray dogs in Nepal have high prevalence of vector-borne pathogens: a molecular survey(Parasites & Vectors, 2020) Díaz-Regañón Fernández, David Rafael; Agulla Pérez, Beatriz; Piya, Bidur; Fernández-Ruiz, Natalia; Villaescusa Fernández, Alejandra; García-Sancho Téllez, Mercedes Guadalupe; Rodríguez Franco, Fernando; Sainz Rodríguez, ÁngelBackground Population of stray dogs is significant in large cities of Nepal, such as Kathmandu. Most of stray dogs suffer a lack of basic health care. Considering the clinical relevance, the broad distribution and the lack of information of canine vector borne diseases (CVBD) in Nepal, the aim of this study was to evaluate the prevalence of different vector-borne pathogens (VBP) in stray dogs living in the metropolitan area of Kathmandu, and to assess different traits as possible risk factors. Methods A total of 70 canine blood samples from stray dogs attended at the Kathmandu Animal Treatment Centre during August 2017 were collected on filter paper (Flinders Technology Associates (FTA) cards). Data regarding signalment, clinical signs and epidemiological characteristics were recorded for each animal. Real-time polymerase chain reaction assays were performed for Leishmania spp., Ehrlichia spp./Anaplasma spp., Babesia spp./Theileria spp. and Hepatozoon canis. Results The overall prevalence detected was 31.43% for Hepatozoon canis, 31.43% for Anaplasma platys, 27.14% for Ehrlichia canis, 18.57% for Leishmania donovani species complex, 12.86% for isolates corresponding to Theileria spp., 12.86% for Babesia vogeli and 2.86% for B. gibsoni. A total of 81.43% of the dogs were positive to at least one of the VBP tested. Co-infections were detected in 41.43% of the dogs. Dogs positive to any of the VBP tested, and particularly to E. canis, were older than those that were negative. Conclusions To our knowledge, this is the first molecular detection of VBP in stray dogs from Kathmandu, Nepal. The high prevalence of VBP detected highlights the need to implement a surveillance programme and control strategies for these CVBD in the population of stray dogs in this area.Item Characterization of the Fecal and Mucosa-Associated Microbiota in Dogs with Chronic Inflammatory Enteropathy(Animals, 2023) Díaz-Regañón Fernández, David Rafael; García-Sancho Téllez, Mercedes Guadalupe; Villaescusa Fernández, Alejandra; Sainz Rodríguez, Ángel; Agulla Pérez, Beatriz; Reyes-Prieto, Mariana; Rodríguez Bertos, Antonio Manuel; Rodríguez Franco, FernandoCanine chronic inflammatory enteropathy implicates multifactorial pathogenesis where immunological dysregulation and gut microbiota changes have a central role. Most sequencing-based taxonomic studies have been focused on the fecal microbiota. However, the analysis of these samples does not provide complete information regarding the composition of the small intestine affected by this canine disease. Therefore, in this study, we aimed to characterize the intestinal bacterial microbiota in dogs with inflammatory bowel disease (IBD) (n = 34) by means of duodenal biopsies and fecal samples collected at the time of the diagnosis and to compare those to a group of healthy dogs (n = 12) using the 16S ribosomal RNA (16S rRNA) gene-targeted sequencing (Illumina MiSeq platform). Our study showed that IBD dogs presented differences in the fecal bacterial communities when compared with healthy dogs, with a lower relative abundance of Prevotellaceae (p = 0.005), Prevotella (p = 0.002), and Prevotellaceae Ga6A1 group (0.006); Erysipelotrichales (p = 0.019), Candidatus Stoquefichus (p < 0.001), Erysipelotrichaceae (p = 0.011), and Allobaculum (p = 0.003); Lachnospiraceae NK4A136 group (p = 0.015), Sellimonas (p = 0.042), Oscillospirales (p = 0.037), Oscillospiraceae UCG–005 (p < 0.001), Faecalibacterium (p = 0.028), and Fournierella (p = 0.034); Acidaminococcales, Acidaminococcaceae, and Phascolarctobacterium (p = 0.001); Aeromonadales (p = 0.026), Succinivibrionaceae (p = 0.037), and Succinivibrio (p = 0.031). On the other hand, a higher relative abundance of Enterococcaceae (Enterococcus; p = 0.003), Streptococcaceae (Streptococcus, p = 0.021), Enterobacterales (p = 0.027), Enterobacteriaceae (p = 0.008), and Escherichia–Shigella (p = 0.011) was detected. Moreover, when evaluating α–diversity, the dogs with IBD showed lower diversity in terms of richness and abundance of species (observed species [p = 0.031] and Shannon index [p = 0.039]). Furthermore, fecal microbiota in dogs with IBD was significantly different from healthy dogs (p = 0.006). However, only a few taxa relative abundance shifts (lower Rubrobacteria, Rubrobacterales, Rubrobacteriaceae, and Rubrobacter [p = 0.002]; Cyanobacteria [p = 0.010], Vampirivibrionia, Obscuribacterales, and Obscuribacteraceae [p = 0.005]; Neisseriaceae [p = 0.004] and Conchiformibius [p = 0.003]) were observed when assessing duodenal-associated microbiota of dogs with IBD. Thus, even if the bowel inflammation mainly affects the small intestine in the IBD-affected dogs of the study, fecal specimens may constitute a better sample due not only to their easy availability but also in terms of searching for bacterial taxa as biomarkers for canine IBD. The use of different diets in the study can also have a partial influence on the microbiota composition. Future studies encompassing multi-omics approaches should evaluate the functionality in both levels to unravel the pathophysiology of canine IBD.Item Changes of Enterocyte Morphology and Enterocyte: Goblet Cell Ratios in Dogs with Protein-Losing and Non-Protein-Losing Chronic Enteropathies(Veterinary Sciences, 2023) Díaz-Regañón Fernández, David Rafael; Gabriel, Vojtech; Livania, Vanessa; Liu, Dongjie; Ahmed, Basant H.; Lincoln, Addison; Wickham, Hannah; Ralston, Abigail; Merodio, María M.; Sahoo, Dipak K.; Zdyrski, Christopher; Meyerholz, David K.; Mochel, Jonathan P.; Allenspach, KarinThis study aimed to assess the morphometry of enterocytes as well as the goblet cell-to-enterocyte ratio in different intestinal segments of dogs with chronic enteropathies (CE). Histopathological intestinal samples from 97 dogs were included in the study (19 healthy juveniles, 21 healthy adults, 24 dogs with protein-losing enteropathy (PLE), and 33 CE dogs without PLE). Healthy adult small intestinal enterocytes showed progressively reduced epithelial cell height in the aboral direction, while juvenile dogs showed progressively increased epithelial cell height in the aboral direction. CE dogs had increased epithelial cell height in the duodenum, while PLE dogs had decreased epithelial cell heights compared to healthy adult dogs. Both the CE and PLE dogs showed decreased enterocyte width in the duodenal segment, and the ileal and colonic enterocytes of CE dogs were narrower than those of healthy adult dogs. CE dogs had a lower goblet cell-to-enterocyte ratio in the colon segment compared to healthy dogs. This study provides valuable morphometric information on enterocytes during canine chronic enteropathies, highlighting significant morphological enterocyte alterations, particularly in the small intestine, as well as a reduced goblet cell-to-enterocyte ratio in the colon of CE cases compared to healthy adult dogs.Item Enfermedad inflamatoria crónica intestinal canina : estudio clínico y caracterización de la microbiota a partir de muestras de heces y biopsias duodenales(2024) Díaz-Regañón Fernández, David Rafael; Rodríguez Franco, Fernando; Villaescusa Fernández, Alejandra; García-Sancho Téllez, Mercedes GuadalupeLa enfermedad inflamatoria crónica intestinal (EICI) es una enfermedad de base inmunomediada caracterizada por la persistencia y recurrencia de signos clínicos gastrointestinales, que constituye una de las causas más frecuentes de vómito y diarrea crónica en la especie canina. La patogenia de la EICI es multifactorial e implica la pérdida de tolerancia a la dieta y los componentes de la microbiota, causando una respuesta inmunitaria exacerbada. El diagnóstico de la enfermedad se alcanza tras realizar un protocolo de exclusión completo de todas las causas que puedan desencadenar un proceso inflamatorio a nivel intestinal y con la evidencia histológica de un infiltrado inflamatorio en la mucosa del intestino delgado, grueso o ambos.La estrecha relación entre el epitelio intestinal del hospedador, el sistema inmunitario y la microbiota, hace que cambios en el ecosistema intestinal que afecten a cualquiera de estos pilares, pueda implicar modificaciones en el resto. En esta línea, y considerando que la microbiota intestinal constituye una compleja población de microorganismos que tienen un papel determinante en el estado de salud del hospedador canino, se ha sugerido que las alteraciones en la microbiota intestinal pueden constituir un potencial biomarcador en la EICI canina. La mayoría de los estudios basados en técnicas de secuenciación realizados hasta la fecha en la especie canina se han centrado en el componente bacteriano de la microbiota fecal. Sin embargo, si la EICI afecta principalmente al intestino delgado, la microbiota asociada a la mucosa intestinal debería mostrar alteraciones más marcadas que la microbiota de muestras fecales. Las heces no proporcionan información completa sobre la potencial presencia de bacterias entero-invasivas o adheridas a la mucosa, ni sobre la composición de la microbiota del intestino delgado. Además, algunos trabajos recientes en medicina humana han mostrado que las alteraciones de la microbiota asociada a la mucosa intestinal presentan una mayor correlación con los índices clínicos de la enfermedad que aquellas observadas en las heces. Este hecho condujo al planteamiento de este trabajo de investigación, cuyo objetivo principal es la caracterización de la microbiota intestinal en la EICI canina, tanto en la mucosa duodenal como en las heces...- Project number: 356
Item Elaboración de material docente para la exploración física del gato(2020) Agulla Pérez, Beatriz; Díaz-Regañón Fernández, David Rafael; García-Sancho Téllez, Mercedes Guadalupe; Rodríguez Franco, Fernando; Sainz Rodríguez, Ángel; Villaescusa Fernández, Alejandra Item Peripheral and intestinal T lymphocyte subsets in dogs with chronic inflammatory enteropathy(Journal of Veterinary Internal Medicine, 2024) Agulla Pérez, Beatriz; Villaescusa Fernández, Alejandra; Sainz Rodríguez, Ángel; Díaz-Regañón Fernández, David Rafael; Rodríguez Franco, Fernando; Calleja Bueno, Lydia; Olmeda, Patricia; García-Sancho Téllez, Mercedes GuadalupeBackground: Dysregulated T lymphocyte response is thought to play a key role inchronic intestinal inflammation (CIE). Objectives: To evaluate the presence of changes in peripheral and intestinal T lym-phocyte subsets and to describe potential immune and inflammatory biomarkers indogs with CIE. Animals: Sixteen healthy dogs and 26 dogs were diagnosed with CIE. Methods: Prospective case-control study evaluating peripheral and intestinal T lym-phocytes using flow cytometry and inflammatory markers obtained from completeblood cell counts. Results: Dogs with CIE had higher peripheral activated T helper (Th) lymphocytes(87/μL [18-273] CIE, 44/μL [16-162] healthy control (HC,P=.013) and regulatory Tcells (Treg; 108/μL [2-257] CIE, 34/μL [1-114] HC,P=.004). In the intestinal epithe-lium, CIE dogs presented lower percentages of Th (4.55% [1.75-18.67] CIE, 8.77%[3.79-25.03] HC,P=.002), activated Th cells (0.16% [0.02-0.83] CIE, 0.33%[0.05-0.57] HC,P=.03) and CD4/CD8 ratio (0.08 [0.02-0.39] CIE, 0.21 [0.07-0.85]HC,P=.003). Conversely, higher percentage of activated T cytotoxic cells (20.24%[3.12-77.12] CIE, 12.32% [1.21-39.22] HC,P=.04) and interferon-gamma (IFN-γ)producing T lymphocytes (7.36% [0.63-55.83] CIE, 1.44% [0.00-10.56] HC,P=.01) within the epithelium was observed. In the lamina propria the percentage of Treglymphocytes was higher (6.02% [1.00-21.48] CIE, 3.52% [0.18-10.52] HC,P=.02). Conclusions and Clinical Importance: Systemic and intestinal immune alterationsoccur in dogs with CIE suggesting that blood IFN-γproducing T lymphocytes and the systemic immune-inflamation index (SII) could potentially serve as biomarkers for the disease.