In the present study, sera from 1146 sheep from eight different locations in the northern provinces of Turkey were investigated against bovine herpesvirus type 1 (BHV-1) by a conventional method, as serum neutralization. Prevalence of antibodies detected against BHV-1 in sheep was found to be 1.74% (20/1146). Serum samples obtained from sheep from 4 different provinces were detected negative against BHV-1. As a result, with this study presence of BHV-1 was detected for the first time in sheep in the northern provinces of
Turkey.Uzorci seruma 1146 ovaca s osam različitih mjesta u sjevernom području Turske bili su pretraženi na goveđi herpesvirus 1 (GHV-1) uobičajenim serum neutralizacijskim testom. Specifična protutijela za GHV-1 dokazana su u 1,74% (20/1146) ovaca. Protutijela za GHV-1 nisu bila dokazana u uzorcima uzetima od ovaca u četirima područjima. Time je prvi put dokazana zaraza tim virusom u ovaca u sjevernim provincijama Turske

Harun Albayrak

Semra Okur-Gumusova

Zafer Yazici

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257ISSN 0372-5480Printed in CroatiaVeterinarski arhiV 77 (3), 257-263, 2007Seroprevalence to bovine herpesvirus type 1 in sheep in TurkeyHarun Albayrak1, Zafer Yazici2*, and Semra Okur-Gumusova2 1Virology Laboratory, Veterinary Control and research institute, Ministry of agriculture, samsun, turkey2Department of Virology, Faculty of Veterinary Medicine, Ondokuz Mayis University, kurupelit, samsun, turkeyAlbAYrAk, H., Z. YAZici, S. Okur-GumuSOvA: Seroprevalence to bovine herpesvirus type 1 in sheep in Turkey. vet. arhiv 77, 257-263, 2007.AbSTrAcTIn the present study, sera from 1146 sheep from eight different locations in the northern provinces of Turkey were investigated against bovine herpesvirus type 1 (BHV-1) by a conventional method, as serum neutralization. Prevalence of antibodies detected against BHV-1 in sheep was found to be 1.74% (20/1146). Serum samples obtained from sheep from 4 different provinces were detected negative against BHV-1. As a result, with this study presence of BHV-1 was detected for the first time in sheep in the northern provinces of Turkey.key words: sheep, bovine herpes virus-1, neutralization, seroprevalence, antibody, TurkeyintroductionBovine herpesvirus-1 (BHV-1) is a member of the Varicellovirus genus, alphaherpesvirinae subfamily, herpesviridae family and has a positive sense double-stranded DNA genome (ALY et al., 2003; MWEENE et al., 2003). The virus creates different infections in ruminants associated with respiratory symptoms such as infectious bovine rhinotracheitis (IBR), genital infection as pustular vulvovaginitis (IPV) and balanoposthitis (IBP) in bulls, conjunctivitis, encephalitis, abortions and fatal multi-systemic infections (MWEENE et al., 2003). BHV-1 can establish latent infections in cattle and this fact serves as a constant source of infection during viral reactivation and re-excretion periods. The major economic losses in the cattle industry occur due to respiratory and reproductive disease caused by this virus (CASTRUCCI et al., 1997). BHV-1 is currently widespread all over the world, for example the USA, Canada, Zaire, Italy, Belgium, India and Turkey (CASTRUCCI et al., 1997; BILGE DAGALP, 1998; *Contact address:Zafer Yazici, DVM, Ph.D, Ondokuz Mayis University, Faculty of Veterinary Medicine, Department of Virology, Kurupelit, 55139, Samsun Turkey, Phone: +90 362 457 60 00 /2811; E-mail: zyazici@omu.edu.tr 258BOELAERT et al., 2000; YESILBAG et al., 2003; RAJKHOWA et al., 2004). The BHV-1 serological surveys with cattle carried out in Turkey ascertained that this virus spreads among dairy and beef cattle in many regions of the country (CABALAR and AKCA, 1994; ALKAN et al., 2000; ALY et al., 2003; AKCA et al., 2004). Previous research by CABALAR and CAN-SAHNA (2000) revealed that the seropositivity rate of BHV-1 was between 20 and 74%. BHV-1 especially infects domestic and wild cattle, and other ruminants may be similarly infected (ANONYM., 2004). In previous studies, cross-species infection bovine and caprine herpesviruses has been demonstrated (LEHMKUHL et al., 1985; YESILBAG et al., 2003). Detection of antibodies against BHV-1 in sheep suggest that sheep may play a role in the epidemiology of BHV-1 (ELAZHARY et al., 1984; JETTEUR et al., 1990; SURESH and SURIBABU, 1993), but sheep do not play a major role in BHV-1 transmission (HAGE et al., 1997).The aim of this study was to investigate the prevalence of BHV-1 in sheep in the northern provinces of Turkey within a broad spectrum for the first time.materials and methodsSheep blood samples (a total of 1146) were collected from animals of both sexes in eight locations in the northern provinces of Turkey and were shipped to our laboratory at 4 °C (Fig. 1). All sampled animals were unvaccinated.H. Albayrak et al.: Seroprevalence to bovine herpesvirus type 1 in sheep in TurkeyFig. 1. Map of Turkey showing the eight locations in the northern provinces of Turkey where serum samples were collected from sheepVet. arhiv 77 (3), 257-263, 2007259Vet. arhiv 77 (3), 257-263, 2007Each blood sample was centrifuged at 3000 rpm for 20 min. at 4º C and sera were obtained. After centrifugation, each serum was poured into a separate tube (Eppendorf, Germany), heat inactivated at 56 °C for 30 min. and stored at -20 ºC until tested. Madin Darby Bovine Kidney cells (MDBK) were used for propagation, titrate and serum neutralization test for BHV-1. Cells were maintained at 37 °C in Dulbecco’s minimal essential medium (DMEM, PAA, Inc., Austria) containing 10% heat-inactivated foetal bovine sera (FCS, PAA Inc., Austria) and DMEM containing 1% foetal serum. In the microneutralization test, DMEM was used with 10% foetal calf serum.BHV-1 (Cooper strain, ATCC Cat. No. VR-864) was used in this study. The infectivity (TCID50) of the virus was determined by the microtitre technique on MDBK cells as described by KAERBER (1964). For investigation of antibody profile against BHV-1, the microneutralization method protocol was used, as previously described by FREY and LIESS (1971). Antibody titer at ½ or greater were accepted as positive.resultsInfectivity of BHV-1 was calculated as 107.0 TCID50 /0.1 mL. The results of antibody profile against BHV-1 from sheep are detailed in Table 1. Overall percentage of BHV-1 in sheep tested was 1.74% (20/1146).Seropositivity rates were 0.51% (1/196), 4.5% (13/288), 2.4% (5/206), 0.58% (1/172) for the locations of Samsun, Tokat, Amasya and Sinop, respectively. All serum samples collected from sheep were negative for the locations of Ordu, Giresun, Rize and Trabzon. Table 1. Seroprevalence of neutralizing antibodies against BHV-1 in sheep sera in eight different locations in northern TurkeyProvince No of sheep tested No of positives (%)Samsun 196 1 (0.51)Tokat 288 13 (4.5)Amasya 206 5 (5.2)Sinop 172 1 (0.58)Ordu 31 -Rize 49 -Giresun 99 -Trabzon 105 -Total 1146 20 (1.74)H. Albayrak et al.: Seroprevalence to bovine herpesvirus type 1 in sheep in Turkey260 Vet. arhiv 77 (3), 257-263, 2007The different age groups of tested animal are detailed in Table 2. Four hundred and eighty of 1146 sheep were aged between 14 and 24 months, and 666 sheep were aged above 24 months. Zero point eight three per cent (4/480) of sheep aged between 14 and 24 months age were found seropositive, while 2.40% (16/666) sheep aged above 24 months were found seropositive for BHV-1 antibodies.Table 2. Distribution of antibodies against BHV-1 in the different age groups of the tested sheepAge No of tested sheep BHV-1 positiveBetween 1 to 2 years 480 4 (0.83%)Above 2 years 666 16 (2.40%)Total 1146 20 (1.74)The antibody titer for BHV-1 ranged between 1/2 and 1/18. Two of twenty positive serum (10%) had titres that ranged between 1/2 to 1/8, and 6 samples (30%) had titres that ranged between 1/32 to 1/64. Twelve (60%) had an antibody titer that ranged between 1/64 to >1/128.DiscussionBHV-1 infects both domestic and wild cattle. Other ruminant species may also be infected with BHV-1 but no other reservoir of BHV-1, apart from ruminants, are known (ANONYM., 2004). Through this, the first wide-ranging study in Turkey, the sero prevalence of BHV-1 in sheep was investigated. As a result, 20 of 1146 (1.74%) sheep were found to be seropositive. The results obtained from this study indicate that BHV-1 infection in sheep exists at low levels in Turkey. In the previous studies, serological evidence of BHV-1 in sheep had been similarly indicated. LEHMKUHL et al. (1985) reported that the prevalence of BHV-1 antibodies among lambs was between 0.4 and 5.4%. ELAZHARY et al. (1984) reported 10.8% BHV-1 seropositivity in tested sheep in Quebec, Canada, while JETTEUR et al. (1990) and CELEDON et al. (2001) detected that 8% tested sera were positive in Zaire and Chile. Prevalence of BHV-1 seropositivity in our study was determined to be between 0.51% and 5.2% depending on the province.Reasons for this seropositivity obtained may be explained by the results of an experimental study by HAGE et al. (1997). In that study it was observed that sheep did not become easily infected with BHV-1 infection, and sheep which possess a latent infection with BHV-1 transferred the agent to cattle during reinfections at a minimal level. In seroprevalence studies performed by ALKAN et al. (1997; 2005) in Turkey, positivity H. Albayrak et al.: Seroprevalence to bovine herpesvirus type 1 in sheep in Turkey261Vet. arhiv 77 (3), 257-263, 2007between 48.3 and 59.7% was determined in the fields we investigated. When the previous studies are compared to our present study, it is evident that BHV-1 infection is encountered rarely in sheep in the area, the reason for this being the restricted behaviour of the BHV-1 host range and the prevention by species barriers. In conclusion, BHV-1 presence in sheep in the northern region of Turkey was determined, although no widespread presence was encountered. referencesAKCA, Y., I. BURGU, S. GUR, S. BILGE-DAGALP (2004): A study on investigations of occurence of some virus infection in Buffaloes in Turkey. Revue Med. Vet. 156, 268-271.ALKAN, F., I. BURGU, S. BILGE-DAGALP, Y. YILDIRIM, A. GENCAY, B. GUNGOR, V. S. ATASEVEN, Y. AKCA (2005): The seroprevalence of BHV-1 infection on selected dairy cattle herds in Turkey. Revue Med.Vet. 156, 166-169.ALKAN, F., A. OZKUL, S. BILGE-DAGALP, K. YESILBAG, T. C. OGUZOGLU, Y. AKCA, I. BURGU (2000): Virological and serological studies on the role of PI-3 virus, BRSV, BVDV and BHV-1 on respiratory infections of cattle:The detection of etiological agents by direct immunofluorescence technique. Dtsch. Tierärztl. Wochenschr. 107, 193-195.ALKAN, F., A. OZKUL, M. T. KARAOGLU, S. BILGE, Y. AKCA, I. BURGU, K. YESILBAG (1997): Siğirlarda viral nedenli solunum sistemi enfeksiyonlarinin seroepidemiyolojisi. A. Ü. Vet. Fak. Derg. 44, 73-80.ALY, N. M., G. G. SHEHAB, I. H. ABD EL-RAHIM (2003): Bovine viral diarrhoea, bovine herpesvirus and parainfluenza-3 virus infection in three cattle herds in Egypt in 2000. Rev. Sci. Tech. 22, 879-892.ANONYMOUS (2004): International Terrestial Animal Health Code. Office International des Epizooties, Paris, http//www.oie.int/BILGE DAĞALP, S. (1998): Kan ve süt serumlarinda IBR-IPV antikorlarinin nötralizasyon testi ile saptanmasi ve süt örneklerinden virus izolasyonu. A. Ü. Vet. Fak. Derg. 45, 313-321.BOELAERT, F., P. BIRONT, B. SOUMARE, M. DISPAS, E. VANOPDENBOSCH, J. P. VERMEERSCH, A. RASKIN, J. DUFEY, D. BERKVENS, P. KERKHOFS (2000): Prevalence of bovine herpesvirus-1 in the Belgian cattle population. Prev. Vet. Med. 45, 285-295.CABALAR, M., K. CAN-ŞAHNA (2000): Doğu ve Güneydoğu Anadolu Bölgesinde süt siğirlarinda Parainfluenza Virus -3, Bovine Herpes Virus -1 ve Respiratory Syncytial Virüs Enfeksiyonlarinin Seroepidemiyolojisi. YYÜ. Vet. Fak. Derg. 11, 101-105.CABALAR, M., Y. AKCA (1994): Fertilite problemli ineklerde infeksiyöz bovine rhinotracheitis, infeksiyöz pustuler vulvovaginitis (IBR/IPV) virus izolasyonu ve seroepidemiyolojisi. A. Ü. Vet. Fak. Derg, 41, 337-349.CASTRUCCI, G., W. B. MARTIN, F. FRIGERI, M. FERRARI, D. SALVATORI, S. TAGLIATI, V. CUTER (1997): Serological survey of herpesvirus 1 infection in selected dairy herds in Northern and Central Italy. Comp. Immunol. Microbiol. Infect. Dis. 20, 315-317.H. Albayrak et al.: Seroprevalence to bovine herpesvirus type 1 in sheep in Turkey262CELEDÓN, M., A. SANDOVA, J. DROGUETT, R. CALFIO, L. ASCENCIO, J. PIZARRO, C. B. NAVARRO (2001): Survey for antibodies to pestivirus and herpesvirus in sheep, goats, alpacas (Lama Pacos), llamas (Lama Glama), guanacos (Lama Guanicoe) and vicuña (Vicugna Vicugna) from Chile. Arch. Med. Vet. 33, 165-172. ELAZHARY, M. A., A. SILIM, S. DEA (1984): Prevalence of antibodies to bovine respiratory syncytial virus, bovine viral diarrhea virus, bovine herpesvirus-1, and bovine parainfluenza-3 virus in sheep and goats in Quebec. Am. J. Vet. Res. 45, 1660-1662.FREY, H. R., B. LIESS (1971): Vermehrungskinetik und verwendbarkeit einer stark cythopathogenen VD-MD virusstammes für diagnostische untersuchugen mit der Mikrotiter Method. Zbl. Vet. Med. B. 18, 61-71.HAGE, J. J., P. VELLEMA, Y. H. SCHUKKEN, H. W. BARKEMA, F. A. M. RIJSEWIJK, J. T. VAN OIRSCHOT, G. H. WENTINK (1997): Sheep do not have a major role in bovine herpesvirus 1 transmission. Vet. Microbiol. 57, 4l-54.JETTEUR, P., E. THIRY, P. P. PASTORET (1990): Serological survey concerning the IBR, CHV2, BVD, PI3, BRS and rinderpest viruses in small ruminants in Zaire. Rev. Elev. Med. Vet. Pays.Trop. 43, 435-437.KAERBER, G (1964): In diagnostic procedures for viral and ricketsial disease. Public Health Ass. New York. 3, 48-50.LEHMKUHL, H. D., R. C. CUTLIP, S. R. BOLIN, K. A. BROGDEN (1985): Seroepidemiologic survey for antibodies to selected viruses in the respiratory tract of lambs. Am. J. Vet. Res. 46, 2601-2604. MWEENE, A. S., H. FUKUSHI, G. S. PANDEY, M. SYAKALIMA, M. SIMUUNZA, M. MALARMO, A. NAMBOTA, K. L. SAMUI, T. TSUBOTA, Y. NAKAZATO, M. ONUMA, J. YASUDA (2003): The prevalance of bovine herpesvirus in traditional cattle in Southern Province, Zambia. Rev. Sci. Tech. 22, 873-877.RAJKHOWA, S., C. RAJKHOWA, H. RAHMAN, K. M. BUJARBARUAH (2004): Seroprevalence of infectious bovine rhinotracheitis in mithun in India. Rev. Sci. Tech. 23, 821-829.SURESH, P., T. SURIBABU (1993): Serological survey of bovine herpesvirus 1 (BHV-l) in sheep in Andrha Pradesh. Ind. J. Anim. Sci. 63, 146-147.YESILBAG, K., S. BILGE-DAGALP, S. OKUR-GUMUSOVA, B. GUNGOR (2003): Studies on herpesvirus infections of goats in Turkey: Prevalence of antibodies to bovine herpesvirus 1. Revue Med. Vet. 154, 772-774.Vet. arhiv 77 (3), 257-263, 2007H. Albayrak et al.: Seroprevalence to bovine herpesvirus type 1 in sheep in TurkeyReceived: 23 February 2006Accepted: 28 May 2007263AlbAYrAk, H., Z. YAZici, S. Okur-GumuSOvA: Seroprevalencija goveđeg herpevirusa 1 u ovaca u Turskoj. vet. arhiv 77, 257-263, 2007.SAŽETAK Uzorci seruma 1146 ovaca s osam različitih mjesta u sjevernom području Turske bili su pretraženi na goveđi herpesvirus 1 (GHV-1) uobičajenim serum neutralizacijskim testom. Specifična protutijela za GHV-1 dokazana su u 1,74% (20/1146) ovaca. Protutijela za GHV-1 nisu bila dokazana u uzorcima uzetima od ovaca u četirima područjima. Time je prvi put dokazana zaraza tim virusom u ovaca u sjevernim provincijama Turske.Ključne riječi: ovce, goveđi herpesvirus 1, neutralizacijski test, seroprevalencija, TurskaVet. arhiv 77 (3), 257-263, 2007H. Albayrak et al.: Seroprevalence to bovine herpesvirus type 1 in sheep in Turkey.

Seroprevalencija goveđeg herpevirusa 1 u ovaca u Turskoj.

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