Detection of bovine respiratory syncytial virus, Pasteurella multocida, and Mannheimia haemolytica by immunohistochemical method in naturally-infected cattle

Turan Yaman 1 , Hüseyin Büyükbayram 2 , Zafer Özyıldız 3 , Funda Terzi 4 , Ahmet Uyar 5 , Ömer Faruk Keles 1 , Şule Yurdagül Özsoy 6 , and Zabit Yener 1
  • 1 Department of Pathology, Faculty of Veterinary Medicine, Van Yuzuncu Yil University, 65090, Van, Turkey
  • 2 Department of Pathology, Faculty of Medicine, Dicle University, 21280, Diyarbakir, Turkey
  • 3 Department of Pathology, Faculty of Veterinary Medicine, Mehmet Akif Ersoy University, 15030, Burdur, Turkey
  • 4 Department of Pathology, Faculty of Veterinary Medicine, Kastamonu University, 37200, Kastamonu, Turkey
  • 5 Department of Pathology, Faculty of Veterinary Medicine, Mustafa Kemal University, 31060, Hatay, Turkey
  • 6 Department of Pathology, Faculty of Veterinary Medicine, Aydın Adnan Menderes University, 09010, Aydın, Turkey


Introduction: The aim of this study was to determine the predisposing effect of bovine respiratory syncytial virus (BRSV) on Pasteurella spp. infection in naturally-induced pneumonia in cattle by immunohistochemical labelling.

Material and Methods: Lungs of cattle slaughtered in the slaughterhouse were examined macroscopically, and 100 pneumonic samples were taken. The samples were fixed in 10% neutral formalin and embedded in paraffin by routine methods. Sections 5 μm in thickness were cut. The streptavidin-peroxidase method (ABC) was used to stain the sections for immuno-histochemical examination.

Results: BRSV antigens were found in the cytoplasm of epithelial cells of bronchi, bronchioles, and alveoles and within inflammatory cell debris and inflammatory exudate in bronchial lumens. Pasteurella spp. antigens were detected in the cytoplasm of the epithelial cells of bronchi and bronchioles, and in cells in the lumens of bronchi and bronchioles. Eleven cases were positive for only one pathogen (six for BRSV and five for Pasteurella spp.), while 35 cases were positive for 2 pathogens: BRSV plus P. multocida (n = 21) or M. haemolytica (n = 14).

Conclusion: The presence of high levels of BRSV in dual infections indicates that BSRV may be the main pneumonia-inducing agent and an important predisposing factor for the formation of Pasteurella spp. infections in cattle naturally afflicted with pneumonia.

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  • 1. Ackermann M.R., Brogden K.A.: Response of the ruminant respiratory tract to Mannheimia (Pasteurella) haemolytica. Microbes Infect 2000, 2, 1079–1088.

  • 2. Ahn K.K., Lee Y.H., Ha Y., Kim D., Chae S., Kim C.H., Chae C.: Detection by in-situ hybridization of Pasteurella multocida toxin (toxA) gene in the lungs of naturally infected pigs. J Comp Pathol 2008, 139, 51–53.

  • 3. Al-Darraji A.M., Cutlip R.C., Lehmkuhl H.D., Graham D.L.: Experimental infection of lambs with bovine respiratory syncytial virus and Pasteurella haemolytica: pathologic studies. Am J Vet Res 1982, 43, 224–229.

  • 4. Al-Darraji A.M., Cutlip R.C., Lehmkuhl H.D.: Experimental infection of lambs with bovine respiratory syncytial virus and Pasteurella haemolytica: immunofluorescent and electron microscopic studies. Am J Vet Res 1982, 43, 230–235.

  • 5. Almeida R.S., Domingues H.G., Coswig L.T., d’Arce R.C.F., de Carvalho R.F., Arns C.W.: Detection of bovine respiratory syncytial virus in experimentally infected balb/c mice. Vet Res 2004, 35, 189–197.

  • 6. Ames T.R.: Dairy calf pneumonia: the disease and its impact. Vet Clin North Am Food Anim Pract 1997, 13, 379–391.

  • 7. Baker J.C.: Bovine respiratory syncytial virus: pathogenesis, clinical signs, diagnosis, treatment, and prevention. Compend Food Anim 1986, 8, 31–38.

  • 8. Belknap E.B., Baker J.C., Patterson J.S., Walker R.D., Haines D.M., Clark E.G.: The role of passive immunity in bovine respiratory syncytial virus-infected calves. J Infect Dis 1991,163, 470–476.

  • 9. Blanco-Viera F.J., Trigo F.J., Jaramillo-Meza L., Aquilar-Romero F.: Serotypes of Pasteurella multocida and Pasteurella haemolytica isolated from pneumonic lesions in cattle and sheep from Mexico. Rev Latinoamericana Microb 1995 37, 121–126.

  • 10. Bryson D.G., Evermann J.F., Liggitt H.D., Foreyt W.J., Breeze R.G.: An immunoperoxidase method of detecting respiratory syncytial virus antigens in paraffin sections of pneumonic bovine lung. Am J Vet Res 1988, 49, 1121–1126.

  • 11. Bryson D.G., McNulty M.S., Logan E.F., Cush P.F.: Respiratory syncytial virus pneumonia in young calves: clinical and pathologic findings. Am J Vet Res 1983, 44, 1648–1655.

  • 12. Ceribasi A.O., Ozkaraca M., Ceribasi S., Ozer H.: Histopathologic, immunoperoxidase, and immunofluorescent examinations on natural cattle pneumonia originated from parainfluenza type 3, respiratory syncytial virus, adenovirus type 3 and herpesvirus type 1. Rev Méd Vét 2014, 165, 201–212.

  • 13. Ceribasi S., Ozkaraca M., Ozer H., Ceribasi A.O.: Diagnosing respiratory syncytial virus using immunofluorescence and immunohistochemistry methods in caprine lungs with bronchopneumonia. Rev Méd Vét 2013, 164, 120–124.

  • 14. Ciszewski D.K., Baker J.C., Slocombe R.F., Reindel J.F., Haines D.M., Clark E.G.: Experimental reproduction of respiratory tract disease with bovine respiratory syncytial virus. Vet Microbiol 1991, 28, 39–60.

  • 15. Dağ S., Gürbüz A., Özen H., Büyük F., Çelebi Ö., Karaman M., Karakurt E.: Immunohistochemical and molecular detection of Mannheimia spp. and Pasteurella spp. in sheep with pneumonia in Kars province, Turkey. Kafkas Univ Vet Fak Derg 2018, 24, 281–288.

  • 16. Dungworth D.L.: The respiratory system. In: Pathology of Domestic Animals, edited by Jubb K.V.F., Kennedy P.C., Palmer N., Academic Press Inc., New York, 1994, pp. 539−699.

  • 17. Frank G.H.: The role of Pasteurella haemolytica in the bovine respiratory disease complex. Vet Med 1986, 12, 841–846.

  • 18. Gershwin L.J.: Bovine respiratory syncytial virus infection: immunopathogenic mechanisms. Anim Health Res Rev 2007, 8, 207–213.

  • 19. Haines D.M., Clark E.G., Chelack B.J.: The detection of bovine respiratory syncytial virus in formalin fixed bovine lung with commercially available monoclonal antibodies and avidin biotin complex immunohistochemistry. Can J Vet Res 1989, 53, 366–368.

  • 20. Haritani M., Ishino S., Oka M., Nakazawa M., Kobayashi M., Narita M., Takizawa T.: Immunoperoxidase evaluation of pneumonic lesions in calves naturally infected with Pasteurella haemolytica. Nippon Juigaku Zasshi 1989, 51, 1137–1141.

  • 21. Haritani M., Nakazawa M., Hashimoto K., Narita M., Tagawa Y., Nakagawa M.: Immunoperoxidase evaluation of the relationship between necrotic lesions and causative bacteria in lungs of calves with naturally acquired pneumonia. Am J Vet Res 1990, 51, 1975–1979.

  • 22. Haritani M., Nakazawa M., Oohashi S., Yamada Y., Haziroglu R., Narita M.: Immunoperoxidase evaluation of pneumonic lesions induced by P. haemolytica in calves. Am J Vet Res 1987, 48, 1358–13562.

  • 23. Haritani M.: Pathological investigations on bovine pneumonic pasteurellosis by use of immunoperoxidase technique. JARQ 1995, 29, 131–136.

  • 24. Haziroglu R., Diker K.S., Gulbahar M.Y., Kul O.: Immunoperoxidase examination of pneumonic bovine lungs naturally infected with Pasteurella haemolytica. Isr J Vet Med 2001, 56, 59–61.

  • 25. Haziroğlu R., Diker K.S., Turkarslan J., Gulbahar M.Y.: Detection of Mycoplasma ovipneumoniae and Pasteurella haemolytica antigens by an immunoperoxidase technique in pneumonic ovine lungs. Vet Pathol 1996, 33, 74–76.

  • 26. Masot A.J., Gomez-Tejedor C., Gazquez A., Redondo E.: Pathological study of experimentally induced bovine respiratory syncytial virus infection in lambs. Zentralbl Veterinarmed B 1996, 43, 233–243.

  • 27. Narita M., Kimura K., Tanimura N., Arai S., Katsuda T.K.: Immunohistochemical characterization of calf pneumonia produced by the combined endobronchial administration of bovine herpesvirus 1 and Pasteurella haemolytica. J Comp Pathol 2000, 123, 126–134.

  • 28. Ozyildiz Z., Tel O.Y., Yilmaz R., Ozsoy S.Y., Keskin, O.: Pathological and microbiological investigations of pneumonic pasteurellosis in sheep. Kafkas Univ Vet Fak Derg 2013, 19, 103–108.

  • 29. Rice J.A., Carrasco-Medina L., Hodgins D.C., Shewen P.E.: Mannheimia haemolytica and bovine respiratory disease. Anim Health Res Rev 2007, 8, 117–128.

  • 30. Sacco R.E., McGill J.L., Palmer M.V., Lippolis J.D., Reinhardt T.A., Nonnecke B.J.: Neonatal calf infection with respiratory syncytial virus: drawing parallels to the disease in human infants. Viruses 2012, 4, 3731–3753.

  • 31. Sacco R.E., McGill J.L., Pillatzki A.E., Palmer M.V., Ackermann M.R.: Respiratory syncytial virus infection in cattle. Vet Pathol 2014, 51, 427–436.

  • 32. Sacco R.E., Nonnecke B.J., Palmer M.V., Waters W.R., Lippolis J.D., Reinhardt T.A.: Differential expression of cytokines in response to respiratory syncytial virus infection of calves with high or low circulating 25-hydroxyvitamin D3. PloS One 2012, 7, e33074.

  • 33. Srikuraman S., Kelling C.L., Ambagala A.: Immune evasion by pathogens of bovine respiratory disease complex. Anim Health Res Rev 2007, 8, 215–229.

  • 34. Tankaew P., Srisawat W., Singhla T., Tragoolpua K., Kataoka Y., Sawada T., Sthitmatee N.: Comparison of two indirect ELISA coating antigens for the detection of dairy cow antibodies against Pasteurella multocida. J Microbiol Methods 2018, 145, 20–27.

  • 35. Thomas L.H., Stott E.J., Jones P.W., Jebbett N.J., Collins A.P.: The possible role of respiratory syncytial virus and Pasteurella spp. in calf respiratory disease. Vet Rec 1980, 107, 304–307.

  • 36. Trigo F.J., Breeze R.G., Liggitt H.D., Evermann J.F., Trigo E.: Interaction of bovine respiratory syncytial virus and Pasteurella haemolytica in the ovine lung. Am j Vet Res 1984, 45, 1671–1678.

  • 37. Yener Z., Ilhan F., Ilhan Z., Saglam Y.S.: Immunohistochemical detection of Mannheimia (Pasteurella) haemolytica antigens in goats with natural pneumonia. Vet Res Commun 2009, 33, 305–313.

  • 38. Zhang W., Liu X., Liu M., Ma B., Xu L., Wang J.: Development of a multiplex PCR for simultaneous detection of Pasteurella multocida, Mannheimia haemolytica, and Trueperella pyogenes. Acta Vet Hung 2017, 65, 327–339.


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