Evaluation of Viral Migration of Different Variants of Equid Alphaherpesvirus 1 in the Central Nervous System of Hamsters by Immunohistochemistry
Animal and Veterinary Sciences
Volume 7, Issue 2, March 2019, Pages: 59-68
Received: Mar. 12, 2019; Accepted: Apr. 12, 2019; Published: Jun. 13, 2019
Views 536      Downloads 140
Aline Aparecida da Silva, Biological Institute, Avenida Conselheiro Rodrigues Alves, São Paulo/SP, Brasil
Elenice Maria Sequetin Cunha, Biological Institute, Avenida Conselheiro Rodrigues Alves, São Paulo/SP, Brasil
Maria do Carmo Custódio de Souza Hunold Lara, Biological Institute, Avenida Conselheiro Rodrigues Alves, São Paulo/SP, Brasil
Eliana Monteforte Cassaro Villalobos, Biological Institute, Avenida Conselheiro Rodrigues Alves, São Paulo/SP, Brasil
Alessandra Figueiredo de Castro Nassar, Biological Institute, Avenida Conselheiro Rodrigues Alves, São Paulo/SP, Brasil
Enio Mori, Pasteur Institute, Avenida Paulista, São Paulo/SP, Brasil
Carolina Natalia Zanuzzi, Department of Histology and Embryology, Faculty of Veterinary Sciences, National University of La Plata, La Plata, Buenos Aires, Argentina
Cecília Mónica Galosi, Department of Histology and Embryology, Faculty of Veterinary Sciences, National University of La Plata, La Plata, Buenos Aires, Argentina
Claudia Del Fava, Biological Institute, Avenida Conselheiro Rodrigues Alves, São Paulo/SP, Brasil
Article Tools
Follow on us
The intranasal inoculation of equid alphaherpesvirus1 (EHV-1) Brazilian variants A4/72, A9/92, A3/97, Iso72/10 and the Argentine variant AR4 in a Syrian hamster model Mesocricetus auratus induced severe encephalitis. Clinical signs included weight loss, lethargy, somnolence, anorexia, and intense salivation two days post-inoculation (dpi), followed by neurological signs such as loss of proprioception, walking in circles, spastic paralysis, seizures, recumbency and death at 3rd dpi (A9/92 and A4/72 variants) and 4th dpi. Respiratory signs such as dyspnea and serosanguinous nasal discharge were also observed. Histopathological changes in brain included mixed inflammatory infiltrate with predominance of mononuclear cells, neuronal degeneration, liquefactive necrosis, hemorrhagic foci, leptomeningitis, perivascular edema, mononuclear infiltration, and perivascular cuffing. Immunohistochemical examination showed viral replication in neurons restrict predominantly to olfactory bulb and frontal cortex (variants AR4 and A3/97) and in groups of cells from distant regions, such as the caudal diencephalon and rostral mesencephalon (variants Iso72/10) and absence of viral antigen labeling of variants A9/92 and A4/72 despite these variants were the most neurovirulent, so new experiments not staggered in days but in hours post inoculation are needed to better understand the viral migration of these variants.
EHV-1, Neurovirulence, Neuropathogenicity, Immunohistochemistry, Myeloencephalopathy
To cite this article
Aline Aparecida da Silva, Elenice Maria Sequetin Cunha, Maria do Carmo Custódio de Souza Hunold Lara, Eliana Monteforte Cassaro Villalobos, Alessandra Figueiredo de Castro Nassar, Enio Mori, Carolina Natalia Zanuzzi, Cecília Mónica Galosi, Claudia Del Fava, Evaluation of Viral Migration of Different Variants of Equid Alphaherpesvirus 1 in the Central Nervous System of Hamsters by Immunohistochemistry, Animal and Veterinary Sciences. Vol. 7, No. 2, 2019, pp. 59-68. doi: 10.11648/j.avs.20190702.15
Copyright © 2019 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Nilsson, M. R.; Corrêa, W. M. Isolamento do vírus do aborto eqüino no Estado de São Paulo. Arq. Inst. Biol. 1966, 33, 23-25.
Laugier, C.; Foucher, N.; Sevin, C.; Leon, A.; Tapprest, J. A 24-Year Retrospective Study of Equine Abortion in Normandy (France). J. Equi. Vet. Sci. 2011, 31, 116-123; DOI: org/10.1016/j.jevs.2010.12.012.
Dunowska, M. A review of Equid herpesvirus 1 for the veterinary practitioner. Part A: clinical presentation, diagnosis and treatment. N. Z. Vet. J. 2014, 62, 171-178.
APHIS (2007). Equine herpesvirus myeloencephalopathy: A Potentially Emerging Disease. Available on line:.(Accessed on 04 January 2016).
Lara, M. C. C. S. H.; Cunha, E. M. S.; Villalobos, E. M. C.; Nassar, A. F. C.; Asano, K. M.; Fernandes, W. R.; Richtzenhain, L. J.; Brandão, P. E.; Mori, E. First isolation of equine Herpesvirus type 1 from a horse with neurological disease in Brazil. Arq. Inst. Biol. 2008, 75, 221-224.
Vandekerckhove, A. P.; Glorieux, S.; Gryspeerdt, A. C.; Steukers, L.; Duchateau, L.; Osterrieder, N.; Van De Walle, G. R.; Nauwynck, H. J. Replication kinetics of neurovirulent versus non-neurovirulent equine herpesvirus type 1 strains in equine nasal mucosal explants. J. Gen. Virol. 2010, 91, 2019-2028; DOI: 10.1099/vir.0.019257-0.
Mori, E.; Borges, A. S.; Delfiol, D. J. Z.; Oliveira Filho, J. P.; Gonçalves, R. C.; Cagnini, D. Q.; Lara, M. C. C. S. H.; Cunha, E. M. S.; Villalobos, E. M. C.; Nassar, A. F. C.; Castro, A. M. M. G.; Brandão, P. E.; Richtzenhain, L. J. First detection of the equine herpesvirus 1 neuropathogenic variant in Brazil. Rev. Sci. Tech. Off. Int. Epiz. 2011, 30, 949-954.
Allen, G. P.; Kydd, J. H.; Slater, J. D.; Smith, K. C. Equid herpesvirus-1 (EHV-1) and -4 (EHV-4) infections. In: Infectious Diseases of Livestock, 2ed. Oxford, 2004; pp 829-859.
Wilson, W. D. Equine herpesvirus 1 myeloencephalopathy. Vet. Clin. North America Equi. Practice. 1997, 13, 53–72.
Schultheiss, P. C.; Collins, J. K.; Hotaling, S. Immunohistochemical demonstration of Equine Herpesvirus-1 antigen in neurons and astrocytes of horses with acute paralysis. Vet. Pathol. 1997, 34, 52-54.
Silva, A. A., Cunha, E. M. S., Lara, M. C. C. S. H., Villalobos, E. M. C., Nassar, A. F. C., Mori, E., Zanuzzi, C. N., Galosi, C. M., and Del Fava, C. Arq. Inst. Biol., 2018, 85, 1-7.
OIE (World Organization For Animal Health) (2015). Equine rhinopneumonitis. OIE Terrestrial Manual. Chapter 2.5.9. p. 897. Available online:.(Accessed on 04 August 2016).
Awan, A. R.; Chong, Y. C.; Field, H. J. The pathogenesis of equine herpesvirus type 1 in the mouse: a new model for studying host responses to the infection. J. Gen. Virol. 1990, 71, 1131-1140.
Gibson, J. S.; Slater, J. D.; Awan, A. R.; Field, H. J. Pathogenesis of equine herpesvirus-1 in specific pathogen-free foals: primary and secondary infections and reactivation. Arch. Virol. 1992, 123, 351–366.
Hasebe, R.; Kimura, T.; Sato, E.; Okazaki, K.; Ochiai, K.; Wada, R.; Ememura, T. Equine herpesvirus-1 induced encephalomyelitis in mice: a comparative study of neuroadapted virus and its parental strain. J. Comp. Path. 2002, 127, 118-125.
Gostonyi, G.; Borchers, K.; Ludwig, H. Pathogenesis of equine herpesvirus-1 infection in the mouse model. APMIS – Acta Path. Microbiol. Immunol. Scandinava. 2009, 117, 10-21; DOI: 10.1111/j.1600-0463.2008.00008.x.
Mori, E.; Mori, C. M. C.; Massironi, S. M. G.; Cunha, E. M. S.; Villalobos, E. M. C.; Lara, M. C. C. S. H.; Fernandes, W. R. Detection of equid herpesvirus 1 DNA by Polymerase Chain Reaction after experimental inoculation of horses with a Brazilian A4/72 strain. Braz. J. Vet. Res. Anim. Sci. 2009, 46, 253-261.
Mori, C. M. C.; Mori, E.; Favaro, L. L.; Santos, C. R.; Lara, M. C. C. S. H.; Villalobos, E. M. C.; Cunha, E. M. S.; Brandão, P. E.; Richtzenhain, L. J.; Maiorka, P. C. Equid Herpesvirus type-1 exhibits neurotropism and neurovirulence in a mouse model. J. Comp. Path. 2012, 146, 202-210; DOI: 10.1016/j.jcpa.2011.04.003.
Smith, K. L.; Allen, G. P.; Branscum, A. J.; Frank Cook, R.; Vickers, M. L.; Timoney, J. P.; Balasuriya, U. B. R. The increased prevalence of neuropathogenic strains of EHV-1 in equine abortions. Vet. Microbiol. 2010, 141, 5–11; DOI: 10.1016/j.vetmic.2009.07.030.
Doll, E. R.; Richards, M. G.; Wallace, M. E. Adaptation of the equine abortion virus to suckling Syrian hamsters. Cornell Vet. 1953, 43, 551-558.
Reiner, U. R.; Lucca Neto, D.; Nilsson, M. R.; Nilsson, T. T.; Kotait, I. Isolamento do vírus do aborto eqüino em Campinas, Estado de São Paulo. In: Congresso Brasileiro de Medicina Veterinária, 13, Brasília. Anais... Brasília: Associação Brasileira de Medicina Veterinária, 1972.
Kotait, I. Herpesvírus eqüino tipo 1: incidência e diagnóstico laboratorial. Rev. Bras. Reprod. Ani. 1991, 1, 164-171.
Cunha, E. M. S.; Peixoto, Z. M. P.; Kroeff, S. S.; Queiroz, L. H.; Kotait, I. Isolamento e identificação do herpesvírus eqüino tipo 1 (EHV-1): confirmação do diagnóstico clínico. In: Reunião Anual do Instituto Biológico, 6 ed, São Paulo, 1993; 15.
Carvalho, R. F.; Spilki, F. R.; Cunha, E. M. S.; Stocco, R. C.; Arns, C. W. Molecular data of UL24 homolog gene (ORF37) from Brazilian isolates of equine herpesvirus type 1. Res. Vet. Sci. 2012, 93, 494-497.
Rollinson, E. A.; White, G. Relative activities of acyclovir and BW759 against Aujeszky’s disease and equine rhinopneumonitis viruses. Antimicrob. Agents Chemother. 1983, 24, 221-226.
Stokes, A.; Allen, G. P.; Pullen, L. A.; Murray, P. K. A hamster model of Equine Herpesvirus Type 1 (EHV-I) infection: passive protection by monoclonal antibodies to EHV-1 glycoproteins 13/14 and 17/18. J. Gen. Virol. 1989, 70, 1173-1183.
Tsujimura, K.; Yamanaka, T.; Kondo, T.; Fukushi, H.; Matsumura, T. Pathogenicity and immunogenicity of equine herpesvirus type 1 mutants defective in either gI or gE gene in murine and hamster models. J. Vet. Medic. Sci. 2006, 68, 1029–1038.
Fukushi, H.; Taniguchi, A.; Yasuda, K.; Yanai, T.; Masegi, T.; Yamaguchi, T.; Hirai, K. A hamster model of equine herpesvirus 9 induced encephalitis. J. Neuro. Virol. 2000, 6, 314-319.
El-Habashi, N.; El-Nahass, E.; Fukushi, H.; Hibi, D.; Sakai, H.; Sasseville, V.; Yanai, T. Experimental intranasal infection of equine herpesvirus 9 (EHV-9) in suckling hamsters: Kinetics of viral transmission and inflammation in the nasal cavity and brain. J. Neuro. Virol. 2010, 16, 242-248; DOI: 10.3109/13550284.2010.489596.
El-Habashi, N.; Murakami, M.; El-Nahass, E.; Hibi, D.; Sakai, H.; Fukushi, H.; Sasseville, V.; Yanai, T. Study on the infectivity of Equine Herpesvirus 9 (EHV-9) by different routes of inoculation in hamsters. Vet. Path. 2011, 47, 1-7; DOI: 10.1177/0300985810375053.
El-Nahass, E.; El-Habashi, N.; Abdelaziz, A. A.; Nayel, M.; Kasem, S.; Fukushi, H.; Tuji, H.; Hirata, A.; Sakai, H.; Yanai, T. Kinetics and pathogenicity of oral infection by Equine Herpesvirus-9 in mice and suckling hamsters. J. Comp. Path. 2012, 146, 211-222; DOI: 10.1016/j.jcpa.2011.05.003.
Mesquita, L. P.; Arévalo, A. F.; Zanatto, D. A.; Miyashiro, S. I.; Cunha, E. M. S.; Souza, M. C. C.; Villalobos, E. M. C.; Mori, C. M. C.; Maiorka, P. C.; Mori, E. Equine herpesvirus type 1 induces both neurological and respiratory disease in Syrian hamsters. Vet. Microbio. 2017, 203, 117–124; DOI: http://dx.doi.org/10.1016/j.vetmic.2017.03.007.
Pagamjav, O.; Sakata, T.; Matsumura, T.; Yamaguchi, T.; Fukushi, H. Natural recombinant between equine herpesviruses 1 and 4 in the ICP4 gene. Microbiol. Immunol. 2005, 49, 167-179.
Nugent, J.; Birch-Machin, I.; Smith, K. C.; Mumford, J. A.; Swann, Z.; Newton, J. R.; Bowden, R. J.; Allen, G. P.; Davis-Poynter, N. Analysis of Equid Herpesvirus 1 strain variation reveals a point mutation of the DNA polymerase strongly associated with neuropathogenic versus non-neuropathogenic disease outbreaks. J. Virol. 2006, 80, 4047–4060; DOI: 10.1128/JVI.80.8.4047-4060.2006.
Goodman, L. B.; Loregian, A.; Perkins, G. A.; Nugent, J.; Buckles, E. L.; Mercorelli, B.; Kydd, J. H.; Palù, G.; Smith, K. C.; Osterrieder, N.; Davis-Poynter, N. A point mutation in a herpesvirus polymerase determines neuropathogenicity. PLoS Pathog. 2007, 11, 1583-1592; DOI: 10.1371/journal.ppat.0030160.
Abo-Sakaya, R.; Ebeid, M. H.; Galila, E. M.; Nayel, M.; Kasem, S.; Abdelwahab, M. G.; Moustafa, A. M.; Arnaout, F. K.; Fukushi, H. D/N752 coding change in DNA polymerase gene (ORF30) plays no role in equid herpesvirus type 1 (EHV-1) growth in vitro. Benha Vet. Med. J. 2014, 26, 1-9.
Kasem, S.; Yu, M. H. H.; Yamada, S.; Kodaira, A.; Matsumura, T.; Tsujimura, K.; Madbouly, H.; Yamaguchi, T.; Ohya, K.; Fukushi, H. The ORF37 (UL24) is a neuropathogenicity determinant of equine herpesvirus 1 (EHV-1) in the mouse encephalitis model. Virology. 2010, 400, 259-270; DOI: 10.1016/j.virol.2010.02.012.
Carvalho, R. F.; Oliveira, A. M.; Souza, A. M.; Passos, L. M.; Martins, A. S. Prevalence of equine herpesvirus type 1 latency detected by polymerase chain reaction. Arch. Virol. 2000, 145, 1773-1787.
Galosi, C. M.; Echeverria, M. G.; Vila Roza, M. V.; Cid De LaPaz, V.; Oliva, G. A.; Etcheverrigaray, M. E. Virus herpes eqüino tipo 1 (EHV-1): patrones de restricción de ADN, perfiles protéicos y estúdios de patogenicidad em ratones. Analecta Vet. 1998, 18, 35-40.
Martín Ocampo, G. P.; Barbeito, C. G.; Galosi, C. M. Genomic Studies and genetic characterization of equid herpesvirus type 1 (EHV-1) strains: estimating the global similarity by distance methods. Int. Res. J. Microbiol. 2011, 2, 079-084.
Crabb, B. S.; Studdert, M. J. Equine herpesvirus 4 (equine rhinopneumonitis virus) and 1 (equine abortation virus). Adv. Virus Res.1995, 45, 135-190.
Allen, G. P. Epidemic disease caused by equine herpesvirus-1:recommendations for prevention and control. Equ.Vet. Educ. 2002, 14, 136-142.
Costa, E. A.; Lima, G. B. L.; Castro, R. T.; Furtini, R.; Portilho, R. V.; Resende, M. Meningo encephalitis in a horse associated with equine herpesvirus 1. Arq. Bras. Med. Vet. Zootec. 2008, 60, 1580-1583.
Pusterla, N.; Hussey, G. S. Equine herpesvirus 1 myeloencephalopathy. Vet. Clin. North Am. Equine Pract. 2014, 30, 489-506; DOI: 10.1016/j.cveq.2014.08.006.
El-Nahass, E.; El-Dakhly, K. M.; El-Habashi, N.; Anwar, S. I.; Sakai, H.; Hirata, A.; Okada, A.; Abo-Sakaya, R.; Fukushi, H.; Yanai, T. Susceptibility of BALB/c-nu/nu mice and BALB/c mice to Equine Herpesvirus 9 infection. Vet. Pathol. 2014, 51, 581-590; DOI: 10.1177/0300985813493932.
Eöry, M. L.; Zanuzzi, C. N.; Fuentealba, N. A.; Sguazza, G. H.; Gimeno, E. J.; Galosi, C. M.; Barbeito, C. G. Effects of different anesthetics in the murine model of EHV-1 infection. Vet. Pathol. 2013, 5, 849-856; DOI: 10.1177/0300985813476062.
Allen, G. P.; Timoney, P. J. Recent advances in our understanding of equine herpesvirus-1 (EHV-1) myeloencephalopathy. In: 107th Annual Meeting of the United States Animal Health Association, 2007, 373–380.
Franz, M.; Goodman, L. B.; VandeWalle, G. R.; Osterrieder, N.; Greenwood, A. D. A Point Mutation in a Herpesvirus Co-Determines Neuropathogenicity and Viral Shedding. Viruses. 2017, 9, 1-7; DOI: 10.3390/v9010006.
Mori, E.; Lara, M. C. C. S. H.; Cunha, E. M. S.; Villalobos, E. M. C.; Mori, C. M. C.; Soares, R. M.; Brandão, P. E.; Fernandes, W. R.; Richtzenhain, L. J. Molecular characterization of Brazilian equid herpesvirus type 1 strains based on neuropathogenicity markers. Braz. J. Microbiol. 2015, 46, 565-570; DOI: 10.1590/S1517-838246220140096.
Cymerys, J.; Słońska, A.; Brzezicka, J.; Tucholska, A.; Chmielewska, A.; Rola, J.; Malik, P.; Bańbura, M. W. Replication kinetics of neuropathogenic and non-neuropathogenic equine herpesvirus type1 (EHV-1) strains in primary murine neurons and ED cell line. Polish. Jour. Vet. Scien. 2016, 4, 777–784; DOI: 10.1515/pjvs-2016-0098.
Perkins, G. A.; Goodman, L. B.; Tsujimura, K.; VandeWalle, G. R.; Kim, S. G.; Dubovi, E. J.; Osterrieder, N. Investigation of the prevalence of neurologic equine herpesvirus type 1 (EHV-1) in a 23-year retrospective analysis (1984–2007). Vet. Microbiol. 2009, 139, 375–378; DOI: 10.1016/j.vetmic.2009.06.033.
Pronost, S.; Léon, A.; Legrand, L.; Fortier, C.; Miszczak, F.; Freymuth, F.; Fortier, G. Neuropathogenic and non-neuropathogenic variants of equine herpesvirus 1 in France. Vet. Microbiol. 2010, 145, 329-33. DOI: 10.1016/j.vetmic.2010.03.031.
Hussey, G. S.; Ashton, L. V.; Quintana, A. M.; VandeWalle, G. R.; Osterrieder, N.; Lunn, D. P. Equine herpesvirus type 1 pUL56 modulates innate responses of airway epithelial cells. Virology. 2014, 464-465, 76-86; DOI: 10.1016/j.virol.2014.05.023.
Holz, C. L.; Nelli, R. K.; Wilson, M. E.; Zarski, L. M.; Azab, W.; Baumgardner, R.; Osterrieder, N.; Pease, A.; Zhang, L.; Hession, S.; Goehring, L. S.; Hussey, S. B.; Hussey, G. S. Viral genes and cellular markers associated with neurological complications during herpesvirus infections. J. Gen. Virol. 2017, 6, 1439-1454; DOI: 10.1099/jgv.0.000773.
Oladunni, F. S.; Sarkar, S.; Reedy, S.; Balasuriya, U. B. R.; Horohov, D. W.; Chambers, T. M. Absence of relationship between type-I interferon suppression and neuropathogenicity of EHV-1. Vet. Immunol. Immunopathol. 2018, 197, 24-30; DOI: 10.1016/j.vetimm.2018.01.007.
Science Publishing Group
1 Rockefeller Plaza,
10th and 11th Floors,
New York, NY 10020
Tel: (001)347-983-5186