ALTERNATIVAS PARA LA PREVENCIN Y EL TRATAMIENTO DEL EXANTEMA COITAL EQUINO
Equine coital exantema (ECE), caused by equid alphaherpesvirus 3 (EHV-3), is a contagious vaenereal disease, characterized by the formation of papules, vesicles, pustules and ulcers on the external genitalia of both mares and stallions. EHV-3 infection does not result in systemic infection, infertil...
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Universidad de Buenos Aires. Facultad de Ciencias Veterinarias
2017
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I28-R145-HWA_7063 |
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dspace |
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Universidad de Buenos Aires |
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I-28 |
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R-145 |
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Repositorio Digital de la Universidad de Buenos Aires (UBA) |
| language |
Español |
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spa |
| topic |
ECE Antivirales TEHU3 Exantema coital equino Etiología Diagnóstico Tratamiento Ciencias Veterinarias |
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ECE Antivirales TEHU3 Exantema coital equino Etiología Diagnóstico Tratamiento Ciencias Veterinarias Vissani, María Aldana ALTERNATIVAS PARA LA PREVENCIN Y EL TRATAMIENTO DEL EXANTEMA COITAL EQUINO |
| topic_facet |
ECE Antivirales TEHU3 Exantema coital equino Etiología Diagnóstico Tratamiento Ciencias Veterinarias |
| description |
Equine coital exantema (ECE), caused by equid alphaherpesvirus 3 (EHV-3), is a contagious vaenereal disease, characterized by the formation of papules, vesicles, pustules and ulcers on the external genitalia of both mares and stallions. EHV-3 infection does not result in systemic infection, infertility and / or abortion. The negative impact of ECE lies on the need to temporarily interrupt reproductive activity,\nthe risk of iatrogenic dissemination of EHV-3, and the occurrence of outbreaks at artificial insemination (AI) and embryo transfer (ET) centers. This may translate into delayed foaling dates, and significant decreases in the number of mares mated and in pregnant rates, as opportunities of being mated are lost. ECE clinical lesions are usually characteristic enough for a clinical diagnosis, but as there are latently infected animals from which the virus can be re-activated and re-excreted without lesions, the identification of these animals is critical to\nprevent transmission, principally from mares to stallions during mating. There are quick diagnostic tests, such as conventional or real time PCR (qPCR), which can be carried out in the laboratory, but these techniques are not applicable for detection of\nexcretion of virus in situ before mating, AI and/or ET. Cessation of breeding of\nclinically affected animals until the end of the excretion period, heightened vigilance\nby the personnel for early recognition of new clinical cases and strict adherence to breeding shed hygiene procedures designed to eliminate mechanical transmission of the virus, are the primary ways to avoid the spread of infection if a case of ECE is observed during the breeding season. Treatment is palliative and is based on the application of antiseptic/astringents, anti-inflammatory agents and broad-spectrum antimicrobials, which contribute to a rapid and uncomplicated healing of genital lesions.The general objective of this work was to decrease the negative impact of ECE infections by the evaluation of antiviral compounds as topical treatment, and the implementation of a rapid detection field assay.\nInitially, the in vitro analysis of four synthetic antiviral compounds (acyclovir,\ncidofovir, ganciclovir and brivudin) and one natural (lamda-carragenan) antiviral\ncompound was carried out. With the results on the 50% effective concentration\n(CE50%) and the CE100%, three concentrations of each of the compounds acyclovir,\ncidofovir and ganciclovir, were selected to continue with the analysis of inhibition of viral production in cell cultures. Next, these three compounds were evaluated against six EHV-3 field strains, isolated in our laboratory between 2001 and 2009. By these assays, we demonstrated that 5 ug/ml of acyclovir, 2 ug/ml of cidofovir and 0.05 ug/ml of ganciclovir are effective to reduce the replication of EHV-3 in vitro, being ganciclovir the most efficient compound.\nWe proceeded then with the in vivo analysis of ganciclovir formulated in base cream at 0.01% p/p (GCV 0.01% p/p), as to be used topically, in a preventive and therapeutic treatment, in mares experimentally infected with EHV-3. While two groups of such mares received a preventive treatment, one of them at 4 h and the other at 4 and 24 h post-infection; a third group received the therapeutic treatment\nduring clinical manifestation of ECE. Two other groups were used as control: one of\nthem did not receive any treatment and the other received base cream without\nantiviral (placebo). To evaluate the in vivo efficacy of GCV 0.01% p/p, the following were assessed: clinical signs and lesions (evaluated by a numerical score); rectal temperature; viral excretion by qPCR and by infection of culture cells; and antibodies response by seroneutralization. In the groups which had received preventive treatment at 4 and 4/24 h (clinically healthy animals, but excreting virus), the clinical severity was significantly reduced, but the excretion of virus was only significantly reduced in one of them (treated 4 h post-infection). Regarding the animals treated during the clinical manifestation of ECE with GCV 0.01% p/p, the clinical severity and the intensity and period of excretion of infective virus (reduced in 8 days) were\nsignificantly reduced. This in vivo experiment represents an advance in the development of a treatment, either preventive or therapeutic, or both, based on the topical administration of ganciclovir to control infection by EHV-3. Adjustments in the concentration, presentation form or frequency of administration, should be\nconsidered in order to achieve complete inhibition of viral excretion.\nFinally, the technological platform known as insulated isothermal PCR (iiPCR),\nwas adopted for the rapid (1:30 h) diagnosis of EHV-3 infection in situ. The validation was carried out to determine the sensibility and specificity, both analytical and diagnostic, compared to qPCR for the detection of EHV-3. Results showed that iiPCR has a 98,2% concordance with qPCR, a technique which it is not practically useful for\nin situ diagnosis before mating. To conclude, both the topical administration of GCV as a treatment for the\ninfection of EHV-3 and the incorporation of the iiPCR technology for the rapid\ndiagnosis in situ, could contribute to decrease the negative impact of ECE in equine production. |
| author2 |
Barrandeguy, María |
| author_facet |
Barrandeguy, María Vissani, María Aldana |
| format |
Tesis doctoral Tesis doctoral acceptedVersion |
| author |
Vissani, María Aldana |
| author_sort |
Vissani, María Aldana |
| title |
ALTERNATIVAS PARA LA PREVENCIN Y EL TRATAMIENTO DEL EXANTEMA COITAL EQUINO |
| title_short |
ALTERNATIVAS PARA LA PREVENCIN Y EL TRATAMIENTO DEL EXANTEMA COITAL EQUINO |
| title_full |
ALTERNATIVAS PARA LA PREVENCIN Y EL TRATAMIENTO DEL EXANTEMA COITAL EQUINO |
| title_fullStr |
ALTERNATIVAS PARA LA PREVENCIN Y EL TRATAMIENTO DEL EXANTEMA COITAL EQUINO |
| title_full_unstemmed |
ALTERNATIVAS PARA LA PREVENCIN Y EL TRATAMIENTO DEL EXANTEMA COITAL EQUINO |
| title_sort |
alternativas para la prevencin y el tratamiento del exantema coital equino |
| publisher |
Universidad de Buenos Aires. Facultad de Ciencias Veterinarias |
| publishDate |
2017 |
| url |
http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=avaposgra&cl=CL1&d=HWA_7063 https://repositoriouba.sisbi.uba.ar/gsdl/collect/avaposgra/index/assoc/HWA_7063.dir/7063.PDF |
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AT vissanimariaaldana alternativasparalaprevencinyeltratamientodelexantemacoitalequino AT vissanimariaaldana alternativasparalaprevencionyeltratamientodelexantemacoitalequino |
| _version_ |
1824356459710775296 |
| spelling |
I28-R145-HWA_70632024-08-27 ALTERNATIVAS PARA LA PREVENCIN Y EL TRATAMIENTO DEL EXANTEMA COITAL EQUINO Equine coital exantema (ECE), caused by equid alphaherpesvirus 3 (EHV-3), is a contagious vaenereal disease, characterized by the formation of papules, vesicles, pustules and ulcers on the external genitalia of both mares and stallions. EHV-3 infection does not result in systemic infection, infertility and / or abortion. The negative impact of ECE lies on the need to temporarily interrupt reproductive activity,\nthe risk of iatrogenic dissemination of EHV-3, and the occurrence of outbreaks at artificial insemination (AI) and embryo transfer (ET) centers. This may translate into delayed foaling dates, and significant decreases in the number of mares mated and in pregnant rates, as opportunities of being mated are lost. ECE clinical lesions are usually characteristic enough for a clinical diagnosis, but as there are latently infected animals from which the virus can be re-activated and re-excreted without lesions, the identification of these animals is critical to\nprevent transmission, principally from mares to stallions during mating. There are quick diagnostic tests, such as conventional or real time PCR (qPCR), which can be carried out in the laboratory, but these techniques are not applicable for detection of\nexcretion of virus in situ before mating, AI and/or ET. Cessation of breeding of\nclinically affected animals until the end of the excretion period, heightened vigilance\nby the personnel for early recognition of new clinical cases and strict adherence to breeding shed hygiene procedures designed to eliminate mechanical transmission of the virus, are the primary ways to avoid the spread of infection if a case of ECE is observed during the breeding season. Treatment is palliative and is based on the application of antiseptic/astringents, anti-inflammatory agents and broad-spectrum antimicrobials, which contribute to a rapid and uncomplicated healing of genital lesions.The general objective of this work was to decrease the negative impact of ECE infections by the evaluation of antiviral compounds as topical treatment, and the implementation of a rapid detection field assay.\nInitially, the in vitro analysis of four synthetic antiviral compounds (acyclovir,\ncidofovir, ganciclovir and brivudin) and one natural (lamda-carragenan) antiviral\ncompound was carried out. With the results on the 50% effective concentration\n(CE50%) and the CE100%, three concentrations of each of the compounds acyclovir,\ncidofovir and ganciclovir, were selected to continue with the analysis of inhibition of viral production in cell cultures. Next, these three compounds were evaluated against six EHV-3 field strains, isolated in our laboratory between 2001 and 2009. By these assays, we demonstrated that 5 ug/ml of acyclovir, 2 ug/ml of cidofovir and 0.05 ug/ml of ganciclovir are effective to reduce the replication of EHV-3 in vitro, being ganciclovir the most efficient compound.\nWe proceeded then with the in vivo analysis of ganciclovir formulated in base cream at 0.01% p/p (GCV 0.01% p/p), as to be used topically, in a preventive and therapeutic treatment, in mares experimentally infected with EHV-3. While two groups of such mares received a preventive treatment, one of them at 4 h and the other at 4 and 24 h post-infection; a third group received the therapeutic treatment\nduring clinical manifestation of ECE. Two other groups were used as control: one of\nthem did not receive any treatment and the other received base cream without\nantiviral (placebo). To evaluate the in vivo efficacy of GCV 0.01% p/p, the following were assessed: clinical signs and lesions (evaluated by a numerical score); rectal temperature; viral excretion by qPCR and by infection of culture cells; and antibodies response by seroneutralization. In the groups which had received preventive treatment at 4 and 4/24 h (clinically healthy animals, but excreting virus), the clinical severity was significantly reduced, but the excretion of virus was only significantly reduced in one of them (treated 4 h post-infection). Regarding the animals treated during the clinical manifestation of ECE with GCV 0.01% p/p, the clinical severity and the intensity and period of excretion of infective virus (reduced in 8 days) were\nsignificantly reduced. This in vivo experiment represents an advance in the development of a treatment, either preventive or therapeutic, or both, based on the topical administration of ganciclovir to control infection by EHV-3. Adjustments in the concentration, presentation form or frequency of administration, should be\nconsidered in order to achieve complete inhibition of viral excretion.\nFinally, the technological platform known as insulated isothermal PCR (iiPCR),\nwas adopted for the rapid (1:30 h) diagnosis of EHV-3 infection in situ. The validation was carried out to determine the sensibility and specificity, both analytical and diagnostic, compared to qPCR for the detection of EHV-3. Results showed that iiPCR has a 98,2% concordance with qPCR, a technique which it is not practically useful for\nin situ diagnosis before mating. To conclude, both the topical administration of GCV as a treatment for the\ninfection of EHV-3 and the incorporation of the iiPCR technology for the rapid\ndiagnosis in situ, could contribute to decrease the negative impact of ECE in equine production. Fil: Vissani, María Aldana. Universidad de Buenos Aires. Facultad de Ciencias Veterinarias. Buenos Aires, Argentina Barrandeguy, María Vissani, María Aldana 2017-12-04 El exantema coital equino (ECE), causado por la infección conalfaherpesvirus equino 3 (Equid alphaherpesvirus 3, EHV-3), es una enfermedad venérea, altamente contagiosa y caracterizada por la formación de pápulas, vesículas, pústulas y úlceras en los genitales externos de yeguas y padrillos. La infección no resulta en enfermedad sistémica, ni tampoco produce infertilidad y/o aborto. El impacto negativo del ECE radica en la necesidad de interrumpir temporalmente la actividad reproductiva; el riesgo de diseminación iatrogénica de\nEHV-3, y la ocurrencia de brotes en centros de inseminación artificial (IA) y transferencia embrionaria (TE). Esto se ve reflejado en retrasos de las fechas de parto, disminuciones significativas del número de yeguas servidas y en las tasas de preñez en las yeguas afectadas, al reducirse las oportunidades de ser servidas. El diagnóstico se realiza en forma clínica, ya que las lesiones son muy características, pero como existen animales patentemente infectados en los que el virus se reactiva y reexcreta en forma subclínica y no predecible, la identificación de los mismos resulta crítica\npara prevenir la transmisión de la infección, principalmente de yeguas a padrillos durante el servicio. Existen técnicas de diagnóstico rápidas, como la PCR convencional y la PCR en tiempo real (qPCR), pero las mismas no son aplicables para determinar la excreción viral subclínica in situ, previo al servicio, IA y/o TE. Las medidas de control frente a brotes de ECE en centros de reproducción son, limitar la diseminación de la infección por medio del reposo sexual en animales clínicamente afectados, detectar tempranamente nuevos casos clínicos, e implementar estrictas medidas de higiene para evitar la transmisión mecánica. El tratamiento es paliativo y consiste en la utilización de antisépticos/astringentes, antiinflamatorios y antimicrobianos, que contribuyen a acortar el período de reposo sexual.\nEl objetivo general de este trabajo fue disminuir el impacto negativo de las infecciones por EHV-3 mediante la evaluación de compuestos antivirales para su uso tópico, y la incorporación de un método rápido de diagnóstico rápido a campo. Inicialmente se realizó un análisis in vitro de cuatro compuestos antivirales sintéticos\n(aciclovir, cidofovir, ganciclovir y brivudin) y uno natural (lamda-carragenano). Con los resultados de concentración efectiva 50 (CE50) y CE100, se seleccionaron tres concentraciones de aciclovir, cidofovir y ganciclovir para continuar con el análisis de\ninhibición de la producción viral en cultivos celulares. Posteriormente, estos tres compuestos fueron evaluados contra seis cepas de campo de EHV-3 aisladas en nuestro laboratorio entre los años 2001 y 2009. Se demostró que 5 ug/ml de aciclovir, 2 ug/ml de cidofovir y 0,05 ug/ml de ganciclovir son efectivos para reducir la replicación de EHV-3 in vitro, siendo ganciclovir el compuesto más eficiente.\nSe continuó entonces, con el análisis de ganciclovir in vivo y el mismo fue preparado en crema base al 0,01% p/p (GCV 0,01% p/p) para su uso tópico, como tratamiento preventivo o terapéutico, en yeguas infectadas experimentalmente con EHV-3. Dos grupos de animales recibieron tratamiento preventivo, uno a las 4 h y el otro a las 4 y 24 h postinfección; mientras que un tercer grupo recibió tratamiento terapéutico con este compuesto una vez iniciado el cuadro clínico. Un grupo de yeguas, experimentalmente infectadas, que no recibió tratamiento antiviral y otro tratado con crema base sin antiviral (placebo) fueron\nlos grupos controles. Como parámetros de la eficacia del compuesto in vivo, se evaluaron: signos clínicos y lesiones (mediante un score numérico); temperatura rectal; excreción viral por qPCR y por infectividad en cultivos celulares; y respuesta de anticuerpos séricos por seroneutralización. En los grupos tratados a las 4 y 4/24 h (animales clínicamente sanos,\npero excretando virus), se redujo significativamente la severidad del cuadro clínico y en uno de ellos se observó una disminución significativa en el tiempo de excreción de virus infectivo (animales tratados a las 4 h post-infección). Con respecto a los animales tratados con GCV 0,01% p/p en forma terapéutica, se observó una reducción de la severidad clínica y una disminución significativa en la intensidad y en el tiempo de excreción de virus infectivo (ocho\ndías menos). Este ensayo in vivo, representa un avance en el desarrollo de un tratamiento preventivo y/o terapéutico, basado en la aplicación tópica de ganciclovir para controlar la\ninfección por EHV-3. Ajustes en la concentración, forma de presentación o frecuencia del tratamiento deberían considerarse para alcanzar una inhibición completa de la excreción\nviral. Por último, se adoptó la plataforma tecnológica conocida como PCR isotérmica aislada\n(iiPCR), para el diagnóstico rápido (1:30 h) de la infección por EHV-3, al pie del animal. Se realizó la validación de este método estableciendo la sensibilidad y especificidad (analítica y\ndiagnóstica), comparado con la qPCR. Los resultados obtenidos con la iiPCR tiene una concordancia de 98,2% con la qPCR, metodología que si bien permitía el diagnóstico rápido de EHV-3 en el laboratorio, no era útil desde el punto de vista práctico para el diagnóstico rápido e in situ, previo a las maniobras reproductivas.\nPara concluir, tanto el uso de GCV en forma tópica como tratamiento de las lesiones producidas por la infección con EHV-3, así como también la incorporación de la tecnología de iiPCR para el diagnóstico rápido al pie del animal, podrían contribuir a disminuir el impacto negativo del ECE en la producción equina. application/pdf ECE Antivirales TEHU3 spa Universidad de Buenos Aires. Facultad de Ciencias Veterinarias info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-ncnd/2.5/ar/ Exantema coital equino Etiología Diagnóstico Tratamiento Ciencias Veterinarias Farmacología y microbiología Doctora de la Universidad de Buenos Aires en Ciencias Veterinarias Alternativas para la prevención y el tratamiento del exantema coital equino info:eu-repo/semantics/doctoralThesis info:ar-repo/semantics/tesis doctoral info:eu-repo/semantics/acceptedVersion http://repositoriouba.sisbi.uba.ar/gsdl/cgi-bin/library.cgi?a=d&c=avaposgra&cl=CL1&d=HWA_7063 https://repositoriouba.sisbi.uba.ar/gsdl/collect/avaposgra/index/assoc/HWA_7063.dir/7063.PDF |