Detección de los herpesvirus equinos 1 y 4 y su relación con transcriptos asociados a la latencia en caballos infectados naturalmente en Colombia

Detection of equine herpesvirus 1 and 4 and its association with latency-associated transcripts in naturally infected horses in Colombia

Objetivo. El objetivo de este estudio fue determinar presencia de anticuerpos y genoma viral de EHV-1 y EHV-4, como también detectar la presencia de transcriptos asociados a latencia (LATs) en una población seleccionada de caballos colombianos. Materiales y métodos. Muestras de suero, nódulos linfáticos submandibulares y ganglio trigémino se obtuvieron de 50 caballos y fueron analizadas. Los sueros se evaluaron para la presencia de anticuerpos contra EHV-1 y EHV-4 mientras que los tejidos se evaluaron inicialmente para la presencia de genoma viral por nPCR. Finalmente, las muestras se emplearon para la detección de LATs a través de RT-PCR. Resultados. En general, 6/50 muestras mostraron anticuerpos para EHV-1 y 44/50 fueron positivos para EHV-4. En cuanto a la detección del genoma viral, 10/50 muestras fueron positivas para EHV-1 y 30/50 fueron positivas para EHV-4; además, 22/35 caballos positivos para DNA de EHV fueron positivos para LATs. El empleo de estas pruebas llevó a ocho posibles combinaciones de resultados. Conclusiones. Se confirma la presencia de estos virus en la población equina colombiana. Las pruebas empleadas demuestran que los caballos pueden tener infección viral simple, co-infecciones con ambos virus, estado de latencia debido a la presencia de los LATs y presencia simultánea de LATs y de replicación de genoma viral en un momento dado. Se aporta al entendimiento del comportamiento de la enfermedad en Colombia y se llama la atención sobre la importancia de implementar diagnósticos complementarios a la serología para el control de estos virus.

1. Patel JR, Heldens J. Equine Herpesvirus 1 (EHV-1) and 4 (EHV-4) epidemiology, disease and immunoprophylaxis: A brief review. Vet J 2005; 170(1):14-23. https://doi.org/10.1016/j.tvjl.2004.04.018
3. Ma G, Azab W, Osterrieder N. Equine herpesvirus type 1 (EHV-1) and 4 (EHV-4) - Masters of co-evolution and a constant threat to equids and beyond. Vet Microbiol 2013; 167(1-2):123-134. https://doi.org/10.1016/j.vetmic.2013.06.018
5. Telford E, Watson M, Perry J, Cullinane A, Davison A. The DNA sequence of equine Herpesvirus-4. J Gen Virol 1998; 79 (5):1197-1203. https://doi.org/10.1099/0022-1317-79-5-1197
7. Osterrieder N, Van de Walle G. Pathogenic potential of equine alphaherpesvirus: The importance of the mononuclear cell compartment in disease outcome. Vet Microbiol 2010; 143 (1):21-28. https://doi.org/10.1016/j.vetmic.2010.02.010
9. Goodman L, Loregian A, Perkins G, Nugent J, Buckles E, Mercorelli B et al. A point mutation in a herpesvirus polymerase determines neuropathogenicity. Plos pathogens 2007; 3(11):1583-1592. https://doi.org/10.1371/journal.ppat.0030160
11. Borchers K, Wolfinger U, Lawrenz B, Schellenbach A, Ludwing H. Equine Herpesvirus 4 DNA in trigeminal ganglia of naturally infected horses detected by direct in situ PCR. J Gen Virol 1997; 78 (5):1109-1114. https://doi.org/10.1099/0022-1317-78-5-1109
13. Baxi M, Efstathion S, Lawrence G, Whalley J, Slater J, Field H. The detection of latency associated transcripts of equine herpesvirus 1 in ganglionic neurons. J Gen Virol 1995; 76 (12):3113-3118. https://doi.org/10.1099/0022-1317-76-12-3113
15. Arthur J, Scarpini C, Connor V, Lachmann R, Tolkovsky A, Efstathiou S. Herpes simplex virus type 1 promotor activity during latency establishment, maintenance, and reactivation in primary dorsal root neurons in vitro. J Virol 2001; 75 (8):3885-3895. https://doi.org/10.1128/JVI.75.8.3885-3895.2001
17. Dunowska M, Gopakumar G, Perrot M, Kendall A, Waropastrakul S, Hartley C, et al. Virological and serological investigation of Equid herpesvirus 1 infection in New Zealand. Vet Microbiol 2015:176(3):219-228. https://doi.org/10.1016/j.vetmic.2015.01.016
19. Borchers K, Wolfinger U, Ludwing H. Latency –associated transcripts of equine Herpesvirus type 4 in trigeminal ganglia of naturally infected horses. J Gen Virol 1999; 80 (8):2165-2171. https://doi.org/10.1099/0022-1317-80-8-2165
21. Pusterla N, Wilson D, Madigan J, Ferraro G. Equine Herpesvirus 1 myeloencephalopathy: A review of recent developments. Vet J 2009; 180 (3):279-289. https://doi.org/10.1016/j.tvjl.2008.08.004
23. Hartley C, Wilks C, Studdert M, Gilkerson J. Comparison of antibody detection assays for the diagnosis of equine herpesvirus 1 and 4 infections in horses. Am J Vet Res 2005; 66(5):921-928. https://doi.org/10.2460/ajvr.2005.66.921
25. Daillo I, Hewitson G, Wright L, kellyM, Rodwell B, Corney B. Multiplex real time PCR for the detection and differentiation of equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4). Vet Microbiol 2007; 123(1):93-103. https://doi.org/10.1016/j.vetmic.2007.02.004
27. Pusterla N, Wilson D, Mapes S, Finno C, Isbell D, Arthur R, Ferraro G. Characterization of viral loads, strain and state of equine herpesvirus-1 using real-time PCR in horses following natural exposure at a racetrack in California. Vet J 2009; 179(2):230-239. https://doi.org/10.1016/j.tvjl.2007.09.018
29. Lunn D, Davis N, Flaminio M, Horohov D, Osterrieder K, Pusterla N, et al. EHV-1 Consensus statement. J Vet Intern Med 2009; 23(3):450-461. https://doi.org/10.1111/j.1939-1676.2009.0304.x
31. Allen G, Bolin D, Bryant U, Carter C, Giles R, Harrison L et al. Prevalence of latent, neuropathogenic equine herpesvirus 1 in the thoroughbred broodmare population of central Kentucky. Equine Vet J 2008; 40(2):105-110. https://doi.org/10.2746/042516408X253127
33. Cano A, Galosi C, Ocampos G, Ramirez G, Vera V, Villamil L, Chaparro J. Equine herpesvirus 1: Characterisation of the first strain isolated in Colombia. Rev sci tech Off int Epiz 2008; 27 (3):893-897. https://doi.org/10.20506/rst.27.3.1846
35. Ruíz J, Góez Y, López A. 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. Rev Colomb Cienc Pecu 2008; 21 (3):372-386.
37. Ruíz J, Góez Y, Urcuqui S, Góngora A, López A. Serologic evidence of equine herpesvirus 1 and 4 infection in two regions of Colombia. Rev Colomb Cienc Pecu 2008; 21 (2):251-258.
39. Borchers K, Slater J. A nested PCR for the detection and differentiation of EHV-1 and EHV-4. J Virol Methods 1993; 45: 331-336. https://doi.org/10.1016/0166-0934(93)90117-A
41. Telford E, Watson M, McBride K, Davison A. The DNA sequence of equine herpesvirus-1. Virology 1992; 189 (1):304-316. https://doi.org/10.1016/0042-6822(92)90706-U
43. Telford E, Watson M, Perry J, Cullinane A, Davison A. The DNA sequence of equine herpesvirus-4. J Gen Virol 1998; 79 (5):1197-1203. https://doi.org/10.1099/0022-1317-79-5-1197
45. Rios P, Benito A, Rivera H. Rinoneumonitis Equina en caballos del valle de Lima. Rev Investig Vet Peru 2002; 13 (2):1609-9117.
47. Ataseven V, Dagalp S, Guzel M, Basaran Z, Tan M, Geraghty B. Prevalence of equine herpesvirus 1 and equine herpesvirus 4 infections in equidae species in Turkey as determined by ELISA and multiplex nested PCR. Res Vet Sci 2009; 86(2):339-344. https://doi.org/10.1016/j.rvsc.2008.06.001
49. Fortier G, Erk E, Fortier C, Richard E, Potier D, Pronost S, et al. Herperviruses in respiratory liquids of horses: Putative implication in airway inflammation and association with cytological features. Vet Microbiol 2009; 139 (1):34-41. https://doi.org/10.1016/j.vetmic.2009.04.021
51. Wilsterman S, Soboll-Hussey G, Lunn D, Ashton L, Callan R, Hussey S et al. Equine herpesvirus -1 infected peripheral blood mononuclear cell subpopulations during viremia. Vet Microbiol 2011; 149 (1):40-47. https://doi.org/10.1016/j.vetmic.2010.10.004
53. Chesters P, Allsop R, Purewal A, Edington N. Detection of latency associated transcripts of equid Herpesvirus 1 in equine leukocytes but not in trigeminal ganglia. J Virol 1997; 71 (5):3437-3443.
55. Slater J, Borchers K, Thackray AM, Field HJ. The trigeminal ganglion is a location for equine herpesvirus 1 latency and reactivation in the horse. J Gen Virol 1994; 75 (8):2007-2016. https://doi.org/10.1099/0022-1317-75-8-2007
57. Carlson J, Traub J, Lunn D, Morley P, Kohler A, Kasper K et al. Equine viral respiratory pathogen surveillance at horse shows and sales. J Equine Vet Sci 2013; 33 (4):229-237. https://doi.org/10.1016/j.jevs.2012.06.006
59. Taouji S, Collobert C, Gicquel B, Saileau C, Brisseau N et al. Detection and isolation of equine herpesviruses 1 and 4 from horses in Normandy: an autopsy study of tissue distribution in relation to vaccination status. J Vet Med B Infect Dis Vet Public Health 2002; 49 (8):394-399. https://doi.org/10.1046/j.1439-0450.2002.00590.x