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Detection of Antinuclear Antibodies in Canine Serum by Immunoperoxidase in MDCK and Indirect Immunofluorescence in HEp2 Cells

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Open Journal of Veterinary Medicine 2023年第5期13卷 41-52页

作者： guillermo valdivia Anda edgar guillermo valdivia lara Ana Cristina Calderón Uribe Alejandra Andrea Escobar Patiño Gloria Elena lara Reyes Laboratorio de Patogenicidad Microbiana Unidad de Investigación Multidisciplinaria FESC UNAM México City México Departamento de Patología Universidad Complutense de Madrid Madrid Espa&#241a Laboratorio DIVeT^® </sup> Estado de México México City México

Antinuclear antibodies are found in animals suffering from Systemic Lupus Erythematosus (SLE) and some other diseases, their presence in the blood is determined by antinuclear antibody (ANA) test using indirect immunofluorescence (IF) with HEp2 cells as a substrate. In this work, an immunoperoxidase (IP) assay was developed to evaluate the ANAs in canine sera, using canine kidney cell lines (MDCK) and compared with a commercial immunofluorescence test on Hep2 cells for this system, a fluoresceinated anti-canine Ig antibody was standardized. The study was performed on 50 sera from dogs submitted to the laboratory with different clinical diagnoses of autoimmune-associated diseases. The procedures on both cells were unified to perform comparisons of the reactions, direct sera or at different dilutions were added to a monolayer of permeabilized MDCK cells, followed by a peroxidized anti-canine IgG conjugate, and a substrate for the IP reaction. The same sera were tested on the commercial IF assay on Hep2 cell system. In 22/50 cases, the presence of LE cells in peripheral blood was determined. A high correlation was found in the detection of antinuclear antibodies between both cell lines and techniques, however there were differences in the reaction patterns in the nucleus and cytoplasm between cell lines. The diffuse nuclear pattern observed in MDCK cells was more related to the presence of high percentages of LE cells in peripheral blood. The differences found in the results were possibly associated with the presence of homologous antigens between the MDCK cells and the dog. In addition, the methodology and standardization for the use and interpretation of a reference serum was developed to unify the interpretation criteria in the laboratory.

关键词： Antinuclear Antibodies Immunoperoxidase in Marbin Darbin Canine Kidney (MDCK) SLE Associate Disease

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Canine Herpesvirus Seroprevalence and Associated Factors in Dogs of Mexico

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Open Journal of Veterinary Medicine 2016年第10期6卷 149-162页

作者： edgar guillermo valdivia lara Jesica Ileana Ángeles Solis Cesar Cuenca Verde Juan Antonio Montaraz Crespo Laura Cobos Marín Juan Carlos Del Río García guillermo valdivia Anda Laboratorio de Patogenicidad Microbiana Unidad de Investigación Multidisciplinaria Facultad de Estudios Superiores Cuautitlán Universidad Nacional Autónoma de México Cuautitlan Izcalli México Laboratorio de Inmunología Facultad de Estudios Superiores Cuautitlán Universidad Nacional Autónoma de México Ciudad de México México Facultad de Medicina Veterinaria Universidad Nacional Autónoma de México Ciudad Universitaria Ciudad de México México Laboratorio de Micotoxinas Unidad de Investigación Multidisciplinaria Facultad de Estudios Superiores Cuautitlán Universidad Nacional Autónoma de México Ciudad de México México

Canine herpesvirus (CHV-1) causes disease associated with high mortality in infect-ed puppies, which represents large financial losses for dog breeders. Since CHV-1 at the time of the study he had not been reported in Mexico, the main objective of this study was to determine the prevalence of antibodies against CHV-1 in canine kennels in the metropolitan area of Mexico City. A commercial enzyme-linked immuno-sorbent assay (ELISA) was used, and the results were compared to those of a viral neutralization test. The ELISA kit uses the complete viral particle as the antigen. The plaque reduction neutralization test was combined with the immunoperoxidase technique because of the low cytopathic effect of CHV-1. Neutralizing antibodies were also detected in 20 randomly selected samples. The prevalence of CHV-1 with ELISA was 87%. The concordance between ELISA and serum neutralization (SN) was 0.1129, the sensitivity of the ELISA against SN was 1.0 (100%), the positive predic-tive value was 0.39 (39%), and the negative predictive value was 1 (100%). These results show that ELISA is useful for monitoring the dog population for CHV-1;a positive test result requires confirmation with an SN test, and a negative ELISA result indicates a high probability of being SN-negative. The only variables that were sta-tistically associated with CHV-1 prevalence were breed and kennel. A statistically significant relationship between the degree of ELISA and SN titer was obtained, with a confidence level of 95%. None of the clinical presentation factors was statistically significant. These results suggest that most of the canine population studied in Mex-ico is in a herpesvirus latency state.

关键词： Canine Herpesvirus Seroprevalence Enzyme-Linked Immunosorbent Assay Serum Neutralization Test

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Stimulation of Phagocytosis and Production of Antibodies against Canine Herpesvirus Type 1 by Pidotimod (Adimod<sup>TM</sup>)

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Open Journal of Veterinary Medicine 2017年第4期7卷 38-47页

作者： edgar G. valdivia lara Andrés ángeles Aldana Jesica I. ángeles Solis Patricia Miranda Castro Juan C. Del Río García guillermo valdivia Anda Laboratory of Microbial Pathogenicity Multidisciplinary Research Unit Faculty of Higher Education Cuautitlán National Autonomous University of Mexico Cuautitlán Izcalli Mexico Laboratory DIVETTM Cuautitlan Izcalli Mexico Biotechnology Laboratory UNAM Cuautitlan Izcalli Mexico Laboratory of Mycotoxins Multidisciplinary Research Unit Faculty of Higher Education Cuautitlan National Autonomous University of Mexico Cuautitlan Izcalli Mexico

Neutrophils are the most important circulating phagocytes. Circulating mono-cytes and precursors of tissue macrophages also have the ability to phagocytize. Pidotimod (ADIMODTM) exerts immunostimulatory and immunoregulatory effects through the stimulation and regulation of cellular immune responses by lymphocytes Canine herpesvirus (CHV) mainly affect puppies between the first and second weeks of age, causing high morbidity in the litter. To date, there is only one commercial vaccine in Europe to prevent disease. In this work, inactivated CHV cultures were inoculated in rabbits, adsorbed and not adsorbed to chitosan nanoparticles. Phagocytosis in the presence or absence of specific antibodies was measured. Response of virus neutralizing antibodies was also evaluated. AdimodTM enhanced the nonspecific and specific phagocytotic response. The association of the virus to the nanoparticles increased the phagocytic ability of blood cells;however, AdimodTM alone had a greater effect on phagocytic activity and generated a stronger immune response that corresponded to the increased phagocytosis (p TM was used.

关键词： Phagocytosis Canine Herpesvirus Pidotimod Stimulation

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Pathology Isolation and Identification of Canine Herpesvirus (CHV-1) in Mexico

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Open Journal of Pathology 2016年第3期6卷 111-121页

作者： E. guillermo valdivia lara Blanca Lilia Barrón Romero Laura Cobos Marín Jessica Ileana Ángeles Solis Sofía González Gallardo Cesar Cuenca Verde Juan Antonio Montaraz Crespo guillermo valdivia Anda Facultad de Estudios superiores Cuautitlán Universidad Nacional Autónoma de México Estado de México México Escuela Nacional de Ciencias Biológicas Instituto Politécnico Nacional Ciudad de México México Universidad Nacional Autónoma de México Ciudad Universitaria Ciudad de México México Facultad de Estudios superiores Cuautitlán Universidad Nacional Autónoma de México Estado de México México

This work presents the pathology description, isolation and identification of canine herpesvirus (CHV-1) in Mexico, a virus that causes a generalized hemorrhagic infection in puppies from the canidae family. Methods: Isolates were obtained from puppies that died within the first four weeks of life and had lesions consistent with canine herpesvirus. Results: The main gross lesions were petechial and ecchymotic hemorrhages in kidneys, liver and lungs;proliferative interstitial nephritis;multifocal necrosis in liver and kidneys;and encephalitis with intranuclear inclusion bodies. Herpesvirus was confirmed through direct immunofluorescence, electron microscopy and polymerase chain reaction for DNA polymerase and glycoprotein B genes. Discussion: Eight strains were isolated and identified as canine herpesvirus corresponding to three of the working cases with gross and microscopic lesions very similar to those described in the literature;then, isolates were confirmed by PCR gene amplification, positive reactions on immunofluorescence and observations from electron microscopy. This work represents the first report of this disease, including gross and histological lesions, and confirmation by isolation and identification of the canine herpesvirus in Mexico.

关键词： Pathology of Puppies Canine Herpesvirus PCR Electron Microscopy

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Stress Biomarkers in the Giant Manta Mobula birostris Associated to Tourism in the Revillagigedo National Park, Mexico

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Open Journal of Veterinary Medicine 2023年第7期13卷 136-146页

作者： Carolina Hernández-Navarro Fernando R. Elorriaga-Verplancken Felipe Galván-Magaña guillermo valdivia-Anda Renato Peña James T. Ketchum edgar M. Hoyos-Padilla Pelagios Kakunjá A.C. La Paz México Centro Interdisciplinario de Ciencias Marinas Instituto Politécnico Nacional La Paz México Facultad de Estudios Superiores Cuautitlán Universidad Nacional Autónoma de México México City México Centro de Investigaciones Biológicas del Noroeste La Paz México MigraMar Bodega Bay California USA Fins Attached: Marine Research and Conservation Colorado Springs USA

A constant increase in dive tourism over the past years in the Revillagigedo National Park, Mexico, could result in a stressful scenario for giant mantas (Mobula birostris). The purpose of this study was to determine the degree of oxidative stress in terms of changes in catalase units (CAT) and muscle glycogen concentration in this species during two periods of different tourism intensity in this protected area. A total of 21 muscle biopsies were collected in March (peak tourism) and November (lower tourism), 2019. Stress biomarkers were analysed by commercial kits from the company Cayman Chemical. Oxidative stress (catalase activity) was significantly higher during the period with lower tourism (p = 0.002), compared to the period with more tourism, suggesting the presence of the general adaptation syndrome. In males, there was a significant difference (p = 0.0005) in oxidative stress between periods of different tourism intensity, suggesting that the reproductive season may be a stressor. Morphotypes showed different oxidative stress (p = 0.031);however, the reason is unknown. No statistical differences were detected in glycogen concentrations between the tourism periods (p = 0.123), probably because this polysaccharide is not a proper indicator of chronic stress in giant mantas. Based on these findings, giant mantas may have an adequate response in terms of oxidative stress due to an increase in tourism;however the observed increase in catalase suggests that it is within the tolerance range of these organisms.

关键词： Catalase Conservation Elasmobranchs Glycogen Oxidative Stress

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