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      Detección de DNA de herpesvirus equino tipos 1 y 4 en mononucleares de sangre periférica y ganglio trigémino de equinos. Infección, latencia y una aproximación a la neuropatogénesis de la cepa circulante Translated title: Equine herpesvirus 1 and 4 DNA detection in peripheral blood mononuclear cells and trigeminal ganglion of equines: Infection, latency and approximation to neuropathogenesis of the strain Translated title: Detecção de DNA tipos de herpesvírus eqüino 1 e 4 nas células mononucleares do sangue periférico e os gânglios trigémeo de cavalos. Infecção, latência e de uma abordagem para o neuropatogénesis da cepa circulante

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          Abstract

          La infección primaria por Herpesvirus Equino tipos 1 y 4 (HVE-1 y HVE-4) se inicia en el tracto respiratorio superior; luego se produce una viremia primaria en la que intervienen linfocitos B y T, la cual le permite al virus alcanzar otros sistemas orgánicos y producir abortos en el último tercio de la gestación, muerte neonatal de potros y síndromes neurológicos, principalmente por causa del HVE-1. Debido al hallazgo de anticuerpos contra HVE-1 y HVE-4 en caballos de dos departamentos de Colombia, el propósito de este estudio fue detectar la presencia del genoma viral en Mononucleares de Sangre Periférica (MNSP) de caballos seropositivos para HVE-1 y HVE-4 y en ganglios trigéminos de equinos en una planta de faenado del departamento de Antioquia. Por medio de una PCR semianidada se amplificaron los genes que codifican por las glicoproteína Hs (gH) de HVE-1 y B (gB) de HVE-4. El 28 y el 19% de los MNSP contenían el gen de la gH y de la gB, respectivamente. En el 57.8 y 47.7% de los ganglios trigéminos evaluados se logró amplificar los genes gH y gB, respectivamente. Para determinar si la cepa de HVE-1 circulante en el departamento de Antioquia poseía potencial neuropatogénico, se amplificó y secuenció el gen de la DNA polimerasa viral, que puede presentar una mutación asociada con neuropatogénesis. Sin embargo, ninguna de las cepas secuenciadas poseía dicha mutación. Los resultados confirman la presencia de la infección por HVE -1 y HVE-4 en el departamento de Antioquia, lo que sugiere que existen animales con infección latente que podrían ser una fuente de infección para otros animales susceptibles.

          Translated abstract

          The infection with Equine Herpesvirus types 1 and 4 (EHV-1 and EHV-4) occurs at the upper respiratory tract. Soon after this takes place a primary cell associated viremia to peripheral blood mononuclear cells (PBMC, mainly on B and T lymphocytes), which allows the virus to reach other organic systems and production of abortions in the last third of gestation, neonatal foal death and neurological syndromes. After primary infection the animals remain latently infected for all the life. Because the presence of antibodies for EHV-1 and EHV-4 in plasma and serum of horses of two departments of Colombia was demonstrated, the objective of the present study as to demonstrate the presence of the viral genome in PBMC from horses diagnosed seropositive for EHV-1 and EHV-4, and in trigeminal ganglion of equines from a slaughterhouse of the Department of Antioquia. By means of a semi-nested PCR, the gene codifying for glycoprotein H (gH) of EHV-1 and gB of EHV-4 were amplified. In PBMC 28 and 19% of gH and of gB amplification were found, respectively; whereas in trigeminal ganglion 57.8 and 47.7% were amplified for gH and gB, respectively. With the aim of assessing whether the circulating strain in the department of Antioquia had a neuropathogenic potential, we amplified and sent to sequencing the gene that encodes the viral DNA polymerase, which could has a mutation that has been associated with neuropathogenic potential. We found that the circulating viral strain in Antioquia does not have such a mutation. The set of our results confirms that infection by EHV is present in the State of Antioquia, Colombia, and that there are equines latently infected which can be a source of infection for other susceptible horses.

          Translated abstract

          A infecção primária por Herpesvirus Eqüino tipo 1 e 4 (HVE-1 e HVE-4) inicia no tracto respiratório superior; depois, há uma viremia primária envolvidos no B e linfócitos T, que permite que o vírus chegue a outros sistemas e produtos biológicos abortos no último terço da gestação, morte neonatal de potros e síndromes neurológicas, devido sobretudo ao facto de HVE-1. Devido à descoberta de anticorpos contra o HVE-1 e HVE-4 cavalos em dois departamentos da Colômbia, O objetivo deste estudo foi o de detectar a presença do genoma viral em MNSP cavalo seropositivos para HVE-1 e HVE-4 4 os gânglios do trigémeo de cavalos em um curativo na planta do departamento de Antioquia. Através de uma PCR semianidada foram amplificados genes codificantes para a glicoproteína Hs (GH) HVE-1 e B (GB) de HVE-4. Em 28 e 19% do MNSP contendo o gene para a GH e no Reino Unido, respectivamente. No 57,8 e 47,7% dos gânglios trigêmeo avaliados foi atingido amplificar genes GH e GB, respectivamente. Para determinar se a estirpe do HVE-1 circulantes no departamento de Antioquia tinha potencial neuropatogénico, foi amplificado e sequenciado o gene da DNA polimerase viral, que pode apresentar uma mutação associada com neuropatogénesis. No entanto, nenhuma das cepas seqüenciadas possuía essa mutação. Os resultados confirmam a presença de infecção e de HVE -1 HVE-4, no departamento de Antioquia, sugerindo que há animais com infecção latente que poderia ser uma fonte de infecção para outros animais sensíveis.

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          Manual of diagnostic tests and vaccines for terrestrial animals

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            Detection of latency-associated transcripts of equid herpesvirus 1 in equine leukocytes but not in trigeminal ganglia.

            Results from Southern hybridization and PCR amplification experiments using a randomly synthesized reverse transcription-PCR product showed that peripheral blood leukocytes from horses showing no clinical signs of disease expressed a putative latency-associated transcript antisense to and overlapping the 3' end of the equid herpesvirus 1 (EHV-1) immediate-early gene (gene 64). A PCR product derived from this transcript has > or =96% identity with the published EHV-1 sequence. In situ hybridization studies of equine bronchial lymph nodes corroborated these findings and are consistent with reactivation data (D. A. Smith, A. Hamblin, and N. Edington, unpublished data), indicating that EHV-1 latency is established predominantly in CD5+/CD8+ leukocytes.
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              The detection of latency-associated transcripts of equine herpesvirus 1 in ganglionic neurons.

              Neural tissues from specific pathogen-free ponies that had been experimentally infected with equine herpesvirus 1 (EHV-1) were analysed by in situ hybridization. Digoxigenin-labelled EHV-1 BamHI fragments spanning almost the entire EHV-1 genome were hybridized to RNA in tissue sections from latently infected trigeminal ganglia. The BamHI E fragment detected EHV-1 RNA antisense to gene 63 (HSV-1 homologue ICP0) in a small number of neurons. Sixteen other BamHI fragments gave negative results in 20 sections tested with each fragment. Latency associated transcripts (LATs) were localized to the neuronal nuclei. EHV-1 nucleotide sequence data in the region reveals the presence of a putative EHV-1 LAT promoter that shares a similar motifs with the HSV-1 LAT promoter, including the LAT promoter-binding factor, and may have a role in EHV-1 LAT expression.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Journal
                rccp
                Revista Colombiana de Ciencias Pecuarias
                Rev Colom Cienc Pecua
                Facultad de Ciencias Agrarias, Universidad de Antioquia (Medellín )
                0120-0690
                September 2008
                : 21
                : 3
                : 372-386
                Affiliations
                [1 ] Universidad de Antioquia Colombia
                [2 ] Universidad Nacional de Colombia Colombia
                Article
                S0120-06902008000300007
                1ece3768-c166-43a4-b525-8de228312413

                http://creativecommons.org/licenses/by/4.0/

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                SciELO Colombia

                Self URI (journal page): http://www.scielo.org.co/scielo.php?script=sci_serial&pid=0120-0690&lng=en
                Categories
                AGRICULTURE, DAIRY & ANIMAL SCIENCE

                Animal agriculture
                herpesvirus pathogenesis,latent infection,viral polymerase sequencing,infecção latente,patogênese por herpesvírus,seqüenciamento de polimerase viral,infección latente,patogénesis por herpesvirus,secuenciación de polimerasa viral

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