Isolation and Molecular Characterization of Equine herpes virus-1 from Horses Localized in Giza Governorate, Egypt | ||||
| Journal of Applied Veterinary Sciences | ||||
| Article 7, Volume 7, Issue 3, July 2022, Page 50-58 PDF (699 K) | ||||
| Document Type: Original Article | ||||
| DOI: 10.21608/javs.2022.134025.1145 | ||||
 View on SCiNiTO | ||||
| Authors | ||||
Hayat S. Emam  1; Abdel Hamid I. Bazid2; Yasser G. M. Abd-Elhafeiz1; Mohammed A. Abo-Elkhair2; Ata E. Beshir3
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| 1Virology Research Unit, Animal Reproduction Research Institute (ARRI), Giza: Egypt | ||||
| 2Department of virology, Faculty of Veterinary medicine,university of Sadat city, Sadat city, Egypt | ||||
| 3Department of Parasitology and Animal Diseases, Veterinary Research Division, National Research Centre, Dokki, Giza, Egypt | ||||
| Abstract | ||||
| Equine herpesvirus-1 (EHV-1) is endemic in many world areas, including Egypt with significant economic losses in the equine industry through causing a high fatality rate among foals associated with respiratory manifestations and abortion in pregnant mares. The present study conducted to investigate the isolation,, molecular characterization, and phylogeny of the circulating EHV-1 isolates among native breeds of horses in Giza governorate. A total of 72 samples (aborted foeti, fetal fluids, placenta, vaginal and nasal swabs) were inoculated into Vero cell culture. The DNA was extracted from the infected tissue culture and used for molecular identification by TaqMan real-time PCR (qRT-PCR), DNA amplification by conventional PCR, and DNA sequencing. The results showed successful isolation of 9 out of 72 tested samples (12.5%) after the third passage in Vero cells with a characteristic cytopathic effect for EHV-1. The obtained results were confirmed by qRT-PCR. The positive samples were subjected to conventional PCR to amplify 869 bp of the glycoprotein B gene. Sequencing and phylogenetic analysis revealed that EHV-1 isolates were closely related to each other and those previously isolated from Egypt and the European EHV-1 strains. In conclusion, the molecular characterization and phylogeny analysis of the circulating EHV-1 improve the understanding of virus epidemiology in Egypt and implicate good control measures. | ||||
| Keywords | ||||
| Equine; Herpes Virus-1; Giza governorate; Molecular characterization; Virus isolation | ||||
| References | ||||
|
abd El-Hafeiz, Y.G., Amal, M.A.M., and Darwish, S.F., 2010. Isolation of Equine Herpesvirus-1( EHV-1) as a cause of reproductive disorders with emphasis on antigenic and genetic identifications. Int. J. Microbiol. Res. 1, 26–32.
Ali, A.A., Refat, N.A., Algabri, N.A., and Sobh, M.S., 2020. Fetal lesions of EHV-1 in equine. Anais Da Academia Brasiliera De Ciencias.92. doi:10.1590/0001-3765202020180837
ALKHALEFA, N. F., ELKON, I.. I., TOLBA, N. A., ELSAYED, E.. L., 2018.Molecular Characterization Of Equine Herpes Virus -1 (Ehv-1) In Egypt. Kafrelsheikh Vet. Med. J. 16, 61–73.
doi:10.21608/kvmj.2018.110208
Ata, E.B., Allam, A.M., Elbayoumy, M.K., and Mahmoud, M.A.E.-F., 2021. Electron microscopy and phylogenetic analysis of Bovine papillomavirus infection in cattle from four Egyptian governorates. Trop. Anim. Health Prod. 53, 160. doi:10.1007/s11250-021-02607-4
Ata, E.B., Ghazy, A.A., and Shaapan, R.M., 2020a. Equine Herpesvirus-1 Infection, Clinical Features, Treatment and Control. Adv. Anim. Vet. Sci. 8, 368–378. doi:10.17582/journal.aavs/2020/8.6.668.679
Ata, E.B., Mahmoud, M.A.E.-F., and Madboli, A.A., 2018a. Molecular detection and immunopathological examination of Deltapapillomavirus 4 in skin and udder of Egyptian cattle. Vet. World 11, 915–920. doi:10.14202/vetworld.2018.915-920
ATA, EB, ZAGHAWA, A., GHAZY, AA, ELSIFY, A., and SHAAPAN, RM, 2018b.Equine Herpes Virus Type-1 Infection: Etiology, Epidemiology, Pathogenesis, Identification and Recent Diagnosis. Asian J. Epidemiol. 11, 34–45. doi:10.3923/aje.2018.34.45
ATA, EBB, SALAMA, A., ZAGHAWA, A., GHAZY, AAA, ELSIFY, A., NAYEL, M., HEGAZY, Y., ABDEL-RAHMAN, E.H.H., and WARDA, S., 2020b.Seroprevalence of equine herpes virus-1 in endemic area of Egypt with risk factors assessment. Bulg. J. Vet. Med. 23, 102–111.doi:10.15547/bjvm.2168
Azab, W., Bedair, S., Abdelgawad, A., Eschke, K., Farag, G.K., Abdel-Raheim, A., Greenwood, A.D., Osterrieder, N., and Ali, A.A.H., 2019. Detection of equid herpesviruses among different Arabian horse populations in Egypt. Vet. Med. Sci. 5, 361–371. doi:10.1002/vms3.176
BRYANT, NA, WILKIE, GS, RUSSELL, CA, COMPSTON, L., GRAFHAM, D., CLISSOLD, L., MCLAY, K., MEDCALF, L., NEWTON, R., DAVISON, AJ., and ELTON, DM, 2018. Genetic diversity of equine herpesvirus 1 isolated from neurological, abortigenic and respiratory disease outbreaks. Transbound. Emerg. Dis. 65, 817–832. doi:10.1111/tbed.12809
Bueno, I., Pearce, P., and Dunowska, M., 2020. Frequency of latent equine herpesvirus type-1 infection among a sample of horses in the central North Island of New Zealand. N. Z. Vet. J. 68, 23–30. doi:10.1080/00480169.2019.1653238
EDINGTON, N., BRIDGES, CG., and PATEL, JR., 1986. Endothelial cell infection and thrombosis in paralysis caused by equid herpesvirus-1: Equine stroke. Arch. Virol. 90, 111–124. doi:10.1007/BF01314149
Elia, G., Decaro, N., Martella, V., Campolo, M., Desario, C., Lorusso, E., Cirone, F., and Buonavoglia, C., 2006. Detection of equine herpesvirus type 1 by real time PCR. J. Virol. Methods 133, 70–75. doi:10.1016/j.jviromet.2005.10.024
Felsenstein, J., 1985. Confidence Limits on Phylogenies: An Approach Using the Bootstrap. Evolution (N. Y). 39, 783. doi:10.2307/2408678
Ghanem, Y.M., Ibrahim, E.-S.M., Yamada, S., Matsumura, T., Osterrieder, N., Yamaguchi, T., and Fukushi, H., 2007. Molecular characterization of the equine herpesvirus 1 strains RacL11 and Kentucky D. J. Vet. Med. Sci. 69, 573–6. doi:10.1292/jvms.69.573
GUO, P.X., GOEBEL, S., PERKUS, M.E., TAYLOR, J., NORTON, E., ALLEN, G., LANGUET, B., DESMETTRE, P., and PAOLETTI, E., 1990. Coexpression by vaccinia virus recombinants of equine herpesvirus 1 glycoproteins gp13 and gp14 results in potentiated immunity. J. Virol. 64, 2399–406. doi:10.1128/JVI.64.5.2399-2406.1990
GUO, X., IZUME, S., OKADA, A., OHYA, K., KIMURA, T., and FUKUSHI, H., 2014. Full genome sequences of zebra-borne equine herpesvirus type 1 isolated from zebra, onager and Thomson’s gazelle. J. Vet. Med. Sci. 76, 1309–12. doi:10.1292/jvms.14-0183
Hassanien, R.T., El-Nahas, E.M., Mahmoud, N.A., and El-Bagoury, G.F., 2020. Molecular characterization of equine herpesvirus-1 and asinine herpesvirus-5 isolated from aborted fetuses of Arabian horses. Thai J. Vet. Med. 50, 397–403.
JELOCNIK, M., NYARI, S., ANSTEY, S., PLAYFORD, N., FRASER, TA, MITCHELL, K., BLISHEN, A., POLLAK, NM, CARRICK, J., CHICKEN, C., and JENKINS, C., 2021. Real-time fluorometric and end-point colorimetric isothermal assays for detection of equine pathogens C. psittaci and equine herpes virus 1: validation, comparison and application at the point of care. BMC Vet. Res. 17, 279. doi:10.1186/s12917-021-02986-8
Kubacki, J., Lechmann, J., Fraefel, C., and Bachofen, C., 2021. Genome Sequence of Equid Alphaherpesvirus 1 (EHV-1) from a Nasal Swab of a Swiss Horse Associated with a Major EHV-1 Outbreak following a Show Jumping Event in Valencia, Spain. Microbiol. Resour. Announc. 10. doi:10.1128/MRA.00732-21
Kumar, S., Stecher, G., and Tamura, K., 2016. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol. Biol. Evol. 33, 1870–1874. doi:10.1093/molbev/msw054
Ma, G., Azab, W., and Osterrieder, N., 2013. Equine herpesviruses type 1 (EHV-1) and 4 (EHV-4)—Masters of co-evolution and a constant threat to equids and beyond. Vet. Microbiol. 167, 123–134. doi:10.1016/j.vetmic.2013.06.018
Maeda, K., Yasumoto, S., Tsuruda, A., Andoh, K., Kai, K., Otoi, T., and Matsumura, T., 2007. Establishment of a novel equine cell line for isolation and propagation of equine herpesviruses. J. Vet. Med. Sci. 69, 989–991. doi:10.1292/jvms.69.989
MAHY, BWJ, and KANGR, HO, 1996. Virological methods manual. Academic press limited, London.
Matumoto, M., Ishizaki, R., and Shimizu, T., 1965. Serologic survey of equine rhinopneumonitis virus infection among horses in various countries. Arch. Gesamte Virusforsch. 15, 609–24. doi:10.1007/BF01245208
MESELHY, NK, ABO EL-KHAIR, MA, AHMED, BM, EL-SOALLY, SA, FARES, AMAM, NAYEL, MA, and HUSSEIN, HA, 2019. Sequence Analysis of Seven Equine Herpes Type 1 Viruses Circulating In Non- vaccinated Arabian and Foreign Horses In Egypt. J. Virol. Sci. 5, 11–21.
OIE, 2018.OIE manual for terrestial animals. Equine rhinopneumonitis (Chapter 2.5.9). http://www.oie.int/standard-se tting/terrestrial-manual/access-online/2.05.09_EQUINE_ RHINO.pdf
OIE, 2017.Equine rhinopneumonitis ( infection with Equid herpes 1 and 4.https://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.05.09_EQUINE_RHINO.pdf
Pavulraj, S., Eschke, K., Theisen, J., Westhoff, S., Reimers, G., Andreotti, S., Osterrieder, N., and Azab, W., 2021. Equine Herpesvirus Type 4 (EHV-4) Outbreak in Germany: Virological, Serological, and Molecular Investigations. Pathog. (Basel, Switzerland) 10. doi:10.3390/pathogens10070810
Pusterla, N., Mapes, S., Wademan, C., White, A., Estell, K., and Swain, E., 2012. Investigation of the role of mules as silent shedders of EHV-1 during an outbreak of EHV-1 myeloencephalopathy in California. Vet. Rec. 170, 465–465. doi:10.1136/vr.100598
Rashwan, Y.S., Alkhalefa, N.F., Nayel, M.A., Bazid, A.-H.I., Badr, Y., Kasem, S.G., El Balal, S., Wasfy, M., and El- Sayed,Magdy, 2019.Molecular detection and phylogenetic analysis of equine herpesviruses associated with respiratory manifestation in Egyptian horses. Biosci. Res. 16, 71.
RECZKO, E., and MAYR, A., 1963. On the fine structure of a virus of the herpes group isolated from horses (short report)]. Arch. Gesamte Virusforsch. 13, 591–3.
Seo, M.-G., Ouh, I.-O., Lee, S.K., Lee, J.-S., Kwon, O.-D., and Kwak, D., 2020. Molecular Detection and Genetic Characteristics of Equine Herpesvirus in Korea. Pathogens 9, 110. doi:10.3390/pathogens9020110
AL-SHAMMARI Z., AHMED B., HAROUN M., AFIFY A., ELSANOUSI A.A., and SHALABY M., 2016. A first molecular phylogeny of an Egyptian equine herpesvirus-4 strain derived from a fetal Arabian horse. Journal of Veterinary Science & Medical Diagnosis 5. https://doi.org/10.4172/2325-9590.1000186
Vargas-Bermudez, D., Corredor F, A., Ramírez-Nieto, G., Vera A, V., and Jaime C, J., 2018. Detection of equine herpesvirus 1 and 4 and its association with latency-associated transcripts in naturally infected horses in Colombia. Rev. MVZ Córdoba 6826–6837. doi:10.21897/rmvz.1371
VAZ, P.K., HORSINGTON, J., HARTLEY, C.A., BROWNING, G.F., FICORILLI, N.P., STUDDERT, MJ, GILKERSON, J.R., and DEVLIN, J.M., 2016. Evidence of widespread natural recombination among field isolates of equine herpesvirus 4 but not among field isolates of equine herpesvirus 1. J. Gen. Virol. 97, 747–755. doi:10.1099/jgv.0.000378
WHALLEY, J.M., ROBERTSON, G.R., SCOTT, N.A., HUDSON, G.C., BELL, C.W., and WOODWORTH, LM, 1989. Identification and nucleotide sequence of a gene in equine herpesvirus 1 analogous to the herpes simplex virus gene encoding the major envelope glycoprotein gB. J. Gen. Virol. 70 ( Pt 2), 383–94. doi:10.1099/0022-1317-70-2-383
Yanni, M. A., Abouelyazeed, E., Ali, H., and Hanna, N., 2021. Verification Of Molecular And Conventional Techniques Used In The Diagnosis Of Equine Herpes Virus In Some Egyptian Governorates. J. Appl. Vet. Sci. 6, 1–8.doi:10.21608/javs.2021.140052
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