Düşük BVDV prevalansı ile seyreden olguların izlenmesinde ekonomik bir izleme tekniği olarak havuzlama ve gerçek zamanlı RT-PZR

Abstract

Bu çalışmanın amacı, havuzlanmış örnek ve gerçek zamanlı RT-PCR kullanımının viremik veya PI hayvanlarda BVDV'yi tespit etmek için uygun olup olmadığının belirlenmesidir. Bu amaçla, Aksaray Üniversitesi Veteriner Fakültesi Teşhis ve Analiz Laboratuvarına bovine viral diarrhoea virus (BVDV) enfeksiyonu açısından test edilmek üzere 62 farklı çiftlikten getirilen, 2701 sığırdan sağlanan kan örneği kullanıldı. Türkiye’nin tüm cografi bölgelerinden örnekleme yapıldı. Kan örnekleri sekizli gruplar halinde havuzlandı, toplam 342 adet havuz oluşturuldu. Her bir havuz BVDV yönünden real time RT-PCR testi ile kontrol edildi. Toplam 342 havuzdan 18 adedinde (%5,26) BVDV antijeni saptandı. Kontrol edilen 62 çiftliğin 18 tanesinde virus tespit edildi. Bu araştırmadan elde edilen en önemli sonuç, hastalık prevalansının düşük (<%10) olduğu durumlarda havuzlama ve real time RT-PCR ile BVDV izlemesinin çok ekonomik olarak yapılabileceğinin gösterilmesidir. Aşılama programlarına başlamak isteyen yetiştiriciler için bir risk / fayda tahmini yapılabilir.

Keywords

• BVDV
• Havuzlama
• Gerçek Zamanlı RT-PZR

BVDV monitoring by pooling and real time RT-PCR as economical monitoring technique with low BVDV prevalence

Abstract

The aim of this study was to determine whether the use of pooled blood samples and real-time RT-PCR are feasible for detecting BVDV in viremic or PI animals. For this purpose, blood samples obtained from 2701 cattle, brought from 62 different farms, were used to test for the presence of bovine viral diarrhoea virus (BVDV) at the Veterinary Virology Department of Aksaray University Faculty of Veterinary Medicine. The sampling was done from all geographical regions in Turkey. Blood samples were pooled in groups of eight, creating a total of 342 pools. Each pool was checked for BVDV with a real-time RT-PCR test. BVDV nucleic acid was detected in 18 (5.26%) of a total of 342 pools. BVDV was detected in 18 of 62 farms. The most important result obtained from this study is that BVDV monitoring by pooling and real time RT-PCR can be done very economically when the disease prevalence is low (<10%). A risk/benefit estimation can be done for breeders who want to start vaccination programs.

Keywords

• BVDV
• Pooling
• Real Time RT-PCR

Supporting Institution

MSD Hayvan Sağlığı, Türkiye (https://www.msd-hayvan-sagligi.com/) ve Diagen Biyoteknolojik Sistemler Sağlık Hizmetleri ve Otomasyonu San. Tic. A.Ş., Ankara (www.diagen.com.tr).

Thanks

This work was supported by MSD Animal Health, Turkey (https://www.msd-hayvan-sagligi.com/) and Diagen Biyoteknolojik Sistemler Sağlık Hizmetleri ve Otomasyonu San. Tic. A.Ş., Ankara (www.diagen.com.tr). The authors would like to acknowledge Ahmet Enis Yağar for his assistance with laboratory procedures.

References

1. Alkan, F., Burgu, İ. (1993) Investigation on the incidence of Bovine Viral Diarrhoea Virus in calves in Turkey. Dtsch Tierarztl Wochenschr, 100:107-109.
2. Ames, T.R. (1986) The causative agent of BVD - its epidemiology and pathogenesis. Veterinary Medicine, 81:848-869.
3. Anonim. (2017) Bovine viral diarrhoea (Chapter 2.4.7), in: Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, http://www.oie.int/international-standard-setting/terrestrial-manual/access-online/, erişim tarihi: 27.01.2018.
4. Avcı, O., Yavru, S. (2013) Konya’da bir süt sığırcılığı işletmesinde doğal enfekte hayvanlarda Bovine Herpesvirus-1 (BHV-1), Bovine Viral Diarrhea Virus (BVDV) ve Bovine Herpesvirus-4 (BHV-4) enfeksiyonlarının araştırılması. Eurasian Journal of Veterinary Science, 29(2):82-86.
5. Baker, J.C. (1995) The clinical manifestations of bovine viral diarrhea infection. Veterinary Clinics of North America-Food Animal Practice, 11:425-445.
6. Blanchard, P.C., Ridpath, J.F., Walker, J.B., Hietala, S.K. (2010) An outbreak of late-term abortions, premature births, and congenital deformities associated with a bovine viral diarrhea virus 1 subtype b induces thrombocytopenia. Journal of Veterinary Diagnostic Investigation, 22(1):128-131.
7. Bolin, S.R. (1995) Control of bovine viral diarrhea infection by use of vaccination. Veterinary Clinics of North America, Food Animal Practice. 11(3): 615-625.
8. Boulard, C., Villejoubert, C. (1991) Use of pooled serum or milk samples for the epidemiological surveillance of bovine hypodermosis. Vet Parasitol, 39:171–183.

9. Bowen, R.A. (2011) Flaviviridae, in: Fenner’s Veterinary Virology, Eds: MacLahlan, NJ and Dubovi EJ, 4th Edition, ss: 467-481, Elsevier Academic Press, Amsterdam.
10. Brock, K.V. (2003) The persistence of bovine viral diarrhea virus. Biologicals, 31:133-135.
11. Brownlie, J. (1991) The pathways for bovine virus diarrhoea virus biotypes in the pathogenesis of disease. Arch Virol. Suppl, 3:79-96.
12. Brownlie, J., Thompson, I., Curwen, A. (2000) Bovine virus diarrhoea virus – strategic decisions for diagnosis and control. In Practice 22(4): 176-187.
13. Burgu, İ., Alkan, F., Özkul, A., Yeşilbağ, K., Karaoğlu, T., Güngör, B. (2003) Türkiye'de süt sığırcılığı işletmelerinde bovine viral diarrhea virus (BVDV) enfeksiyonunun epidemiyolojisi ve kontrolü. Ankara Üniv Vet Fak Derg, 50:127-133.
14. Cowling, D.W., Gardner, I.A., Johnson, W.O. (1999) Comparison of methods for estimation of individuallevel prevalence based on pooled samples, Preventive Veterinary Medicine, 39: 211-225.
15. Çabalar, M., Karaoğlu, T. (1999) Sığırlarda BVD Virus enfeksiyonuna karşı antikor varlığının araştırılmasında Nötralizasyon İmmunperoksidaz (NPLA) ve Serum Nötralizasyon (SN) testlerinin karşılaştırılması. Ankara Üniv Vet Fak Derg, 46:249-255.
16. Dorfman, R. (1943) The detection of defective members of large populations. Annals of Mathematical Statistics, 14(4):436–440. https://doi.org/10.1214/aoms/1177731363.
17. Dubovi, E.J. (1992) Genetic diversity and BVD virus. Comp. Immun. Microbiol. Infect. Dis. 15(3): 155-162.
18. Duman, R., Yavru, S., Kale, K., Avcı, O. (2009) Seroprevalence of viral upper respiratory infections in dairy cattle. Kafkas Univ Vet Fak Derg, 15(4):539-542.
19. Edwards, S. (1990) The diagnosis of bovine virus diarrhoea mucosal disease in cattle. Rev Sci Tech, 9:115-130.
20. Furstenau, T.N., Cocking, J.H., Hepp, C.M., Fofanov, V.Y. (2020) Sample pooling methods for efficient pathogen screening: Practical implications, PLoS ONE 15(11):e0236849. doi.org/10.1371/journal.pone.0236849.
21. Grooms, D.L. (2004) Reproductive consequences of infection with bovine viral diarrhea virus. Vet Clin North Am Food Anim Pract, 20:5-19.
22. Gunn, G.J., Saatkamp, H.W., Humphry, R.W., Stott, A.W. (2005) Asessing economic and social pressure fort he control of bovine viral diarrhoea virus. Prev Vet Med, 72:149-162.
23. Hanon, J.B., De Baere, M., De la Ferté, C., Roelandt, S., Van der Stede, Y., Cay, B. (2017) Evaluation of 16 commercial antibody ELISAs for the detection of bovine viral diarrhea virus–specific antibodies in serum and milk using well-characterized sample panels. Journal of Veterinary Diagnostic Investigation, 29(6):833-843.
24. Houe, H. (1999) Epidemiological features and economical importance of bovine virus diarrhoea virus (BVDV) infections. Veterinary Microbiology, 64(2-3):89-107.
25. Houe, H. (2003) Economic impact of BVDV infection in dairies. Biologicals, 31:137-143. https://doi.org/10.1016/S1045-1056(03)00030-7.
26. Kennedy, J.A., Mortimer, R.G., Powers, B. (2006) Reverse transcription-polymerase chain reaction on pooled samples to detect bovine viral diarrhea virus by using fresh ear-notch-sample supernatants. J Vet Diagn Invest. 18(1):89-93. doi: 10.1177/104063870601800113.
27. Kline, R.L., Brothers, T.A., Brookmeyer, R., Zeger, S., Quinn, T.C. (1989) Evaluation of human immunodeficiency virus seroprevalence in population surveys using pooled sera. J. Clin. Microbiol. 27: 1449-1452.
28. Koonin, E.V., Dolja, V.V., Krupovic, M., Varsani, A., Wolf, Y.I., Yutin, N., Zerbini, F.M., Kuhn, J.H. (2020) Global organization and proposed megataxonomy of the virus world. Microbiol Mol Biol Rev. Mar 4;84(2):e00061-19. doi: 10.1128/MMBR.00061-19.
29. Lanyon, S.R., Anderson, M.L., Reichel, M.P. (2014) Pooling serum to identify cohorts of nonmilking cattle likely to be infected with Bovine viral diarrhea virus by testing for specific antibodies. Journal of Veterinary Diagnostic Investigation, 26 (3):346–353, doi: 10.1177/1040638714526596.
30. Laureyns, J., Ribbens, S., de Kruif, A. (2010) Control of bovine virus diarrhoea at the herd level: Reducing the risk of false negatives in the detection of persistently infected cattle. Vet J, 184(1):21-26.
31. Lindberg, A.L.E. (2003) Bovine viral diarrhoea virus infections and its control: A review. Veterinary Quarterly, 25(1):1-16. Mars, M.H., Van Maanen, C. (2005) Diagnostic assays applied in BVDV control in the Netherlands. Prev Vet Med, 72(1-2):43-48.
32. McArthur, M.J. (2004) Foetal protection provided by viracare 3 vaccine in seronegative heifers when challenged with a new zealand strain of bovine viral diarrhoea virus, Proc 34th Annual Seminar, Society of Sheep & Beef Cattle Veterinarians NZVA, 2004 © VetLearn® Foundation (NZ).
33. McGowan, M.R, Kirkland, P.D., Rodwell, B.J., Kerr, D.R., Carroll, C.L. (1993) A field investigation of the effects of bovine viral diarrhea virus infection around the time of insemination on the reproductive performance of cattle. Theriogenology, 39:443-9.
34. Moennig, V., Becher, P. (2018) Control of bovine viral diarrhea. Pathogens, doi:10.3390/pathogens7010029. Moerman, A., Straver, P., de Jong, M., Quak, J., Baanvinger, T., van Oirschot, J. (1993) A long term epidemiological study of bovine viral diarrhoea infections in a large herd of dairy cattle. Veterinary Record, 132:622-626.
35. Muñoz-Zanzi, C.A, Johnson, W.O., Thurmond, M.C., Hietala, S.K. (2000) Pooled-sample testing as a herd-screening tool for detection of bovine viral diarrhea virus persistently infected cattle, J VET Diagn Invest,12: 195, doi: 10.1177/104063870001200301.
36. Neill, J.D., Ridpath, J.F., Fischer, N., Grundhoff, A., Postel, A., Becher, P. (2014) Complete genome sequence of pronghorn virus, a pestivirus. Genome Announc, Jun 12;2(3):e00575-14. doi: 10.1128/genomeA.00575-14.
37. Nelson, D.D., Duprau, J.L., Wolff, P.W., Evermann, J.F. (2015) Persistent bovine viral diarrhea virus ınfection in domestic and wild small ruminants and camelids ıncluding the mountain goat (Oreamnos americanus). Front Microbiol, Vol 6: Article No. 01415, doi: 10.3389/fmicb.2015.01415.
38. Newcomer, B.W., Walz, P.H., Givens, M.D., Wilson, A.E. (2015) Efficacy of bovine viral diarrhea virus vaccination to prevent reproductive disease: a meta-analysis. Theriogenology, 83:360-365.
39. Okur, G.S., Yazıcı, Z., Albayrak, H., Çakıroğlu, D. (2007) Seroprevalence of bovine viral respiratory diseases. Acta Veterinaria (Beograd), 57(1):11-16.
40. Olafson, R., MacCallum, A.D., Fox, F.H. (1946) An apparently new transmissible disease of cattle. Cornell Vet, 36:205-213.
41. Öncül, S., Meriç, İ., Korkut, F. (1964) Türkiye’de ilk defa Lalahan Zootekni Araştırma Enstitüsü sığırlarında tespit edilen Mucosal Disease’in klinik yönü. Lalahan Zoo Arş Enst Derg, 4:186-189.
42. Patel, J.R., Shiletto, R.W., Williams, J., Alexander, D.C.S. (2002) Prevention of transplacental infection of bovine foetus by bovine viral diarrhoea virus through vaccination. Archives of Virology. 147:2453-2463.
43. Peterhans, E., Jungi, T.W., Schweizer, M. (2003). BVDV and innate immunity. Biologicals, 31:107-112.
44. Polak, M.P., Antos, A., Rola, J., Żmudziński, J.F. (2016) Viral shedders in a herd vaccinated against infection with bovine viral diarrhoea virus (BVDV) without prior testing for the presence of persistently infected animals, J Vet Res 60:379-384, doi: 10.1515/jvetres-2016-0056.
45. Richter, V., Lebl, K., Baumgartner, W., Obritzhauser, W., Käsbohrer, A., Pinior, B. (2017) A systematic worldwide review of the direct monetary losses in cattle due to bovine viral diarrhoea virus infection. Vet J, 220:80-87. https://doi.org/10.1016/j.tvjl.2017.01.005.
46. Ridpath, J. (2010) Bovine Viral Diarrhea Virus: Global status. Vet Clin North Am Food Anim Pract, 26(1):105-121.
47. Rodake, L., Granatova, M., Vesely, T., Nevorankova, Z. (1997) Monoclonal antibody for the demonstration by ELISA of antibodies to protein p24 of enzootic bovine leukosis virus in individual and pooled blood serum and milk samples. J Vet Med, B 44:425–436.
48. Rypuła, K., Płoneczka-Janeczko, K., Bania, J., Wałecka, E., Bierowiec, K., Rozpędek, W. (2013) Reduction of prevalence of persistent BVDV infection in cattle herds by long-term vaccination program (preliminary clinical study). Polish Journal of Veterinary Sciences, 16:381-383.
49. Sarıkaya, B., Azkur, A.K., Gazyagci, S., Aslan, M.E. (2012) Genetic variability of bovine viral diarrhea virus in the 5’-UTR in the Central Anatolia of Turkey, Acta Scientiae Veterinariae. 40(1):1013.
50. Smith, R.L., Sanderson, M.W., Walz, P.H., Givens, M.D. (2008) Sensitivity of polymerase chain reaction for detection of bovine viral diarrhea virus in pooled serum samples and use of pooled polymerase chain reaction to determine prevalence of bovine viral diarrhea virus in auction market cattle. J Vet Diagn Invest, 20:75-78.
51. Stahl, K., Alenius, S. (2012) BVDV control and eradication in Europe-an update. Jpn J Vet Res, 60:31-39.
52. Stokstad, M., Loken, T. (2002) Pestivirus in cattle: Experimentally induced persistent infection in calves. J Vet Med, 49:494-501.
53. Şimşek, A. (1997) Investigation of persistent bovine viral diarrhea virus infection in clinically healthy cattle herds and its epizootiologic importance. Vet Bil Derg, 13:113-119.
54. Şimşek, Ş., Gürçay, M., Parmaksız, A., İçen, İ., Sekin, S., Koçhan, A., Çelik, Ö.Y., Çakmak, F. (2017) Diyarbakır yöresindeki sığırların sindirim ve solunum sistemi problemlerinde enzootik bovine leukosis (ebl), bovine viral diare (bvd), ınfeksiyöz bovine rhinotracheitis (ibr) ve mavi dil (bt) enfeksiyonlarının rollerinin araştırılması. Dicle Üniv Vet Fak Derg, 10(1):13-18.
55. Tan, M.T., Karaoğlu, T., Erol, N., Yıldırım, Y. (2006) Serological and virological investigations of bovine viral diarrhoea virus (BVDV) infection in dairy cattle herds in Aydın province. Turk J Vet Anim Sci, 30:299-304.
56. Timurkan, M.O., Aydın, H. (2019) Increased genetic diversity of BVDV strains circulating in Eastern Anatolia, Turkey: first detection of BVDV-3 in Turkey. Tropical Animal Health and Production, 51(7):1953-1961.
57. Yeşilbağ, K., Alpay, G., Tuncer, P. (2012) Bir süt sığırcılığı işletmesinde bovine viral diarrhoea (bvd) virus enfeksiyonunun kontrol ve eliminasyonu. Uludag Univ J Fac Vet Med, 31(1):11-17.
58. Yeşilbağ, K., Förster, C., Özyiğit, M.O., Alpay, G., Tuncer, P., Thiel, H.J., König, M. (2014) Characterisation of bovine viral diarrhea virus (BVDV) isolates from an outbreak with haemorrhagic enteritis and severe pneumonia. Vet Microbiol, 169(1-2): 42-49.
59. Yılmaz, H., Altan, E., Ridpath, J., Turan, N. (2012) Genetic diversity and frequency of bovine viral diarrhea virus (BVDV) detected in cattle in Turkey. Comp Immunol Microbiol Infect Dis, 35(5):411-416.
60. Yılmaz, V. (2016) Prevalence of antibodies to bovine viral diarrhea virus (BVDV) in blood and milk serum in dairy cattle in Kars district of Turkey. Indian J. Anim. Res, 50(5):811-815.