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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9 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 Epidemiological study of hemotropic mycoplasmas (hemoplasmas) in cats from central Spain(Parasites & Vectors, 2018) Díaz-Regañón Fernández, David Rafael; Villaescusa Fernández, Alejandra; Ayllón Santiago, Tania; Rodríguez Franco, Fernando; García-Sancho Téllez, Mercedes Guadalupe; Agulla Pérez, Beatriz; Sainz Rodríguez, ÁngelHemotropic mycoplasmas (hemoplasmas) have been found infecting cats worldwide. However, studies about feline hemoplasma infections in Spain are scarce. Therefore, the purpose of the research was to evaluate the prevalence of feline hemotropic mycoplasmas and to characterize risk factors and clinical findings associated with these infections in a cat population from the Madrid area, Spain. Methods Polymerase chain reaction (PCR) was employed to detect Mycoplasma haemofelis (Mhf), “Candidatus Mycoplasma haemominutum” (CMhm) and “Candidatus Mycoplasma turicensis” (CMt) in blood samples from 456 client-owned and 138 stray cats from Madrid. In order to assess associations between these hemoplasma infections and epidemiological parameters, data regarding signalment, environment, prophylaxis measures, retrovirus status, clinical signs and laboratory findings were compiled, whenever possible. Results DNA of feline hemoplasmas was detected from the blood of 63 out of 594 cats (10.6%), with a prevalence of 3.7% (22/594) for Mhf, 8.1% (48/594) for CMhm and 0.5% (3/594) for CMt. Stray cats had statistically higher prevalences of feline hemoplasmas (15.9%) and, specifically, of Mhf (8.7%) than client-owned cats (9 and 2.2%, respectively). A total of seven cats (1.17%) were co-infected with “Candidatus M. haemominutum” and M. haemofelis, two (0.33%) with “Candidatus M. haemominutum” and “Candidatus M. turicensis” and another one (0.17%) with M. haemofelis and Candidatus “M. turicensis”. Male gender, collection of blood during warm months and FeLV/FIV positivity status were associated with hemotropic mycoplasma infection in cats from Madrid. Additionally, within the group of client-owned cats, hemoplasma infection was associated with adult age, outdoor access, and the existence of low haematocrit, erythrocyte count and haemoglobin concentration values. Conclusions To our knowledge, this is the first epidemiological survey of feline hemoplasmas performed in central Spain (Madrid). Our study confirms that “Ca. Mycoplasma haemominutum”, Mycoplasma haemofelis and “Ca. Mycoplasma turicensis” are infecting client-owned and stray cats in this region of Spain, “Ca. Mycoplasma haemominutum” being the most prevalent species. More studies are necessary to help understand the role of the natural infection by these species of hemoplasma in cats.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 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.- 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.Item Twenty-year evolution of Leishmania infantum infection in dogs in Valdeorras (Galicia, Northwestern Spain): implication of climatic factors and preventive measures(Parasites and Vectors, 2024) Olmeda García, Patricia; Díaz-Regañón Fernández, David Rafael; Villaescusa Fernández, Alejandra; Amusátegui Amusátegui, Inmaculada; García, Adolfo; Herrero, Francisco; Tesouro, Miguel A.; Rodríguez Franco, Fernando; García-Sancho Téllez, Mercedes Guadalupe; Martín Fraile, Daniel; Sainz Rodríguez, ÁngelBackground: Abiotic factors play a significant role in the evolution of Leishmania infantum infection due to its vectorial nature. This study aims to assess the evolution in the detection of new L. infantum infection cases in Valdeorras (Ourense, Northwestern Spain) over a 20-year period and how different climatic variables and preventive measures may have affected it. Methods: Indirect immunofluorescence antibody tests (IFAT) were performed on serum samples collected from dogs attending the ‘Servicios Veterinarios de Sil’ veterinary clinic (Valdeorras, Northwestern Spain) between May 2003 and April 2023 to detect L. infantum exposure. The percentage of new cases of L. infantum infection was calculated from May of one year to April of the following year. Climatic conditions in the region, global sales of ectoparasiticides and the number of vaccines against L. infantum delivered in the veterinary clinic from 2003 to 2022 were recorded. Statistical analyses were conducted to determine the associations between these factors and the percentage of new cases of L. infantum infection. Results: A total of 2909 dogs were assessed, and 3785 IFAT tests were performed between May 2003 and April 2023. The mean percentage of new seropositive cases over the 20-year period studied was 21.65 ± 10.8%, with a decline from the beginning to the end of the period studied. The percentage was significantly higher between May 2003 and April 2008 compared with the other periods (May 2008 to April 2013, May 2013 to April 2018 and May 2018 to April 2023). There was a positive correlation between the percentage of new cases of L. infantum infection and the maximum relative humidity in winter. Conversely, there was a negative correlation between the percentage of new cases and sales of ectoparasiticides and vaccination against L. infantum. Conclusions: This study is one of the longest evaluations of the evolution of L. infantum infection in a fixed location and its association with external factors including climatic conditions and preventive measures. The results confirm that Valdeorras is a high-risk area for L. infantum infection. The use of ectoparasiticides and vaccines against L. infantum has been shown to play a significant role in preventing L. infantum infection, highlighting the crucial role of veterinarians in the fight against this disease.