Villaescusa Fernández, Alejandra

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First Name
Last Name
Villaescusa Fernández
Universidad Complutense de Madrid
Faculty / Institute
Medicina y Cirugía Animal
Medicina y Cirugía Animal
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Now showing 1 - 10 of 10
  • Publication
    Elaboración de material docente para la exploración física del gato
    (2020-06-17) Agulla Pérez, Beatriz; Díaz-Regañón Fernández, David Rafael; García-Sancho Téllez, Mercedes Guadalupe; Rodríguez Franco, Fernando; Sainz Rodríguez, Ángel; Villaescusa Fernández, Alejandra
  • Publication
    Evaluación del inmunofenotipo en la especie canina: estudio del efecto de la administración de doxiciclina en perros sanos y con ehrlichiosis
    (Universidad Complutense de Madrid, Servicio de Publicaciones, 2011-08-09) Villaescusa Fernández, Alejandra; Tesouro Diez, Miguel Ángel; Sainz Rodríguez, Ángel
  • Publication
    Isolation and immunophenotyping by flow cytometry of canine peripheral blood and intraepithelial and lamina propria duodenal T lymphocytes
    (Elsevier, 2021-07-28) 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, Alejandra
    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.
  • Publication
    Characterization of the Fecal and Mucosa-Associated Microbiota in Dogs with Chronic Inflammatory Enteropathy
    (MDPI, 2023-01-17) 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, Fernando
    Canine 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.
  • Publication
    Epidemiological study of hemotropic mycoplasmas (hemoplasmas) in cats from central Spain
    (Springer Nature, 2018-03-20) Díaz-Regañón Fernández, David Rafael; Villaescusa Fernández, Alejandra; Ayllón Santiago, Tania; Rodríguez Franco, Fernando; García-Sancho Téllez, Mercedes Guadalupe; Agulla Pérez, Beatriz; Sainz Rodríguez, Ángel
    Hemotropic 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.
  • Publication
    Stray dogs in Nepal have high prevalence of vector-borne pathogens: a molecular survey
    (Springer Nature, 2020-04-21) 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, Ángel
    Background 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.
  • Publication
    Effect of chemically modified tetracycline-8 (CMT-8) on hematology, blood chemistry, cytokines and peripheral blood lymphocyte subsets of healthy dogs
    (Elsevier, 2021-05) Sainz Rodríguez, Ángel; García-Sancho Téllez, Mercedes Guadalupe; Rodríguez Franco, Fernando; San Andrés Larrea, Manuel Ignacio; Rodríguez Fernández, Casilda; Lucas Burneo, José Julio De; San Andrés Larrea, María Dolores De; Agulla Pérez, Beatriz; Villaescusa Fernández, Alejandra
    Tetracyclines are antibiotics widely used in human and veterinary medicine. Effects on the immune system and inflammatory response, including effects on blood leukocytes proliferation and function and in cytokines synthesis, have been described. Chemically modified tetracyclines (CMT) have lost their antimicrobial activity, but maintain these other properties. This study analyzes the effect of chemically modified tetracycline-8 (CMT-8) on the evolution of complete blood count, blood chemistry, the mRNA expression of selected cytokines and peripheral blood lymphocyte subpopulations distribution in healthy dogs. CMT-8 at a dose of 10 mg/kg once daily was administered per os to six healthy dogs. A control group of five healthy dogs, living in the same conditions than dogs treated with CMT-8, received placebo with an identical therapeutic regimen. When given at the doses used in this study, no side effects of CMT-8 were detected, suggesting a good tolerance and a limited toxicity of the drug. Dogs treated with CMT-8 showed a gradual increase in mean corpuscular hemoglobin. The administration of CMT-8 in healthy dogs did not affect blood mRNA expression of IFN-γ, TNFα, IL-4, IL-6, IL-10, IL-12 p40 and IL-13. However, the lymphocytes expressing class II MHC on their surface decreased during the first two weeks of CMT-8 treatment and subsequently increased for the next three months. Considering the absence of antimicrobial properties of the drug, the effects of CMT-8 detected in this study seem to be unrelated to the classical antimicrobial activity attributed to tetracyclines.
  • Publication
    Atlas interactivo de ecografía reproductiva bovina
    (2015) Re, Michela Tatiana; Blanco Murcia, Francisco Javier; Gaspar Simón, Ignacio De; Villaescusa Fernández, Alejandra
  • Publication
    Elaboración de material docente para la enseñanza de la ecografía pulmonar en el bovino
    (2022) Re, Michela Tatiana; Villaescusa Fernández, Alejandra; Sánchez Calabuig, María Jesús; Coronel Araujo, Carlos Críspulo; Romero Crespo, Isabel; Mejías López, Elena; Blanco Murcia, Francisco Javier
    Con este proyecto se quiere desarrollar material audiovisual que sirva de apoyo a las prácticas que reciben los alumnos de Veterinaria. Dicho material incluirá descripciones de la técnica ecográfica pulmonar y de la interpretación de las imágenes, incluyendo material visual de tipo multimedia sobre la ecografía del aparato respiratorio fisiológico y patológico en el bovino. Con este proyecto se pretende facilitar el acceso a material docente útil para el aprendizaje de la ecografía, de forma que los estudiantes y los profesionales veterinarios puedan afianzar sus conocimientos de dicha técnica de forma previa a su empleo en los animales vivos.
  • Publication
    Elaboración de material docente para la enseñanza de la exploración física del caballo
    (2022-02-14) Fores Jackson, Paloma; Gutiérrez Cepeda, Luna; Villaescusa Fernández, Alejandra; Villalba Orero, María; Ruiz de León Robledo, María de los Ángeles; Crespo Castejón, Francisco; Dominuez Gimbernat, Monica
    El objetivo principal del presente proyecto fue incorporar una serie de recursos educativos virtuales que facilitaran e implementaran la enseñanza de la exploración física del caballo.