Research Article
BibTex RIS Cite

Solunum Bozukluğu olan Sığırlarda Bulaşıcı Sığır Pleuropnömonisine Karşı Oluşan Hümoral İmmün Yanıtın Western Blot Tekniği, Kompleman fiksasyon testi ve Kompetatif Enzyme Linked İmmunosorbent Assay Testi ile Değerlendirilmesi

Year 2021, , 113 - 117, 31.12.2021
https://doi.org/10.47027/duvetfd.995044

Abstract

Bu çalışmada, solunum bozuklukları gösteren sığırlardan alınan 253 serum numunesi, doğrulayıcı olarak Western Immunoblotting test (IBT), Kompleman fiksasyon testi (CFT) ve kompetatif enzyme linked immunosorbent assay (c-ELISA) ile test edildi. CFT ile 253 serum örneğinden ikisi (%0,8) pozitif, ikisi (%0,8) şüpheli, c-ELISA ile 7 (%2,8) serum numunesi pozitif ve 25'i (%9,9) şüpheli bulundu. Öte yandan, 110, 98, 95, 62/60 ve 48 kDa' olarak tanımlanması gereken antijenik bantlar, serum örneklerinin hiçbirinde tespit edilmedi. Bazı serum numunelerinin İmmünoblot analizi sonucunda bir veya üç arasında bant görüldü ve bu bantların bazıları oldukça soluktu.
MmmSc suşları arasındaki elektroforetik profiller ile ilgili benzerlikleri tespit etmek için sığırlardan izole edilen Mycoplasma türlerinin tam hücre proteinleri ve MmmSC referans suşları (Mycoplasma mycoides subsp. mycoides SC Botswana (Afrika suşu), Mycoplasma mycoides subsp. mycoides SC PG1 (referans suş), Mycoplasma mycoides subsp. mycoides SC V5 (Australian strain), M. bovigenitalium, M. bovirhinis, M. capricolum subsp. capricolum, M.canadense, M.alkalescens and M. bovis). sodium dodecyl sulphate polyacrilamide gel elektroforezi (SDS-PAGE). ile analiz edildi
The result of this analysis indicated that antigens of 220, 78, 82, 68, 24 kDa were common among all tested strains. According to these results, it was thought that positive results by both CFT and c-ELISA, also non specific band patterns observed by IBT might be caused by a cross reaction between tested mycoplasma strains. As conclusion, it is recommended that all the positive results by CFT and c-ELISA be confirmed by IBT.
Bu analizin sonucu, 220, 78, 82, 68, 24 kDa antijenlerinin test edilen tüm suşlar arasında yaygın olduğunu gösterdi. Bu sonuçlara göre, CFT ve c-ELISA pozitif sonuçlarının ve IBT ile gözlemlenen spesifik olmayan bant örneklerinin, test edilen mikoplazma suşları arasındaki çapraz reaksiyonlardan kaynaklanabileceği düşünüldü., CFT ve c-ELISA ile elde edilen tüm pozitif sonuçların IBT testi ile doğrulanması gerektiği sonucuna varıldı.

References

  • 1. Nicholas RA, Bashiruddin JB, Ayling RD, Miles RJ (2000). Contagious bovine pleuropneumonia: a review of recent developments. Vet Bull. 70:827-838.
  • 2. OIE. (2015). Contagious bovine pleuropneumonia. In: Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Parisi
  • 3. Wawegama NK., & Browning GF (2017). Improvements in diagnosis of disease caused by Mycoplasma bovis in cattle. Animal Production Science. 57(7): 1482-1487.
  • 4. Amanfu W. (2019) Contagious Bovine Pleuropneumonia. In: Kardjadj M., Diallo A., Lancelot R. (eds) Transboundary Animal Diseases in Sahelian Africa and Connected Regions. Springer, Cham.
  • 5. Nicholas RAJ, Santini FG, Clark KM, Palmer NM, Santis P De, Bashiruddin JB (1996). A comparison of serological tests and gross lung pathology for detecting contagious bovine pleuropneumonia in two groups of Italian cattle, Vet Rec. 139: 89-93,
  • 6. Regalla J, Gonçalves R, Ribeiro N, Duarte L, Penha Gonçalves A (1996). Use of immunoblotting test for carrier state definition in contagious bovine pleuropneumonia, In Mycoplasmas of ruminants: pathogenicity, diagnostics, epidemiology and molecular genetics, Ed. J. Frey and K . Sarris, Rep, No. EUR 16934 EN, European Commission, Luxembourg, 72-74.
  • 7. Regalla J, Gonçalves R, Niza Ribeiro J, Duarte L, Nicholas R, Bashiruddin J, De Santis P, Garrido Abellan F, Penha Gonçalves A (1999). Development of immunoblotting as a diagnostic tool for contagious bovine pleuropneumonia. In, COST 826 Agriculture and biotechnology “Mycoplasmas of ruminants: pathogenicity, diagnostics, epidemiology and molecular genetics”, (Ed. D. Bergonier, X. Berthelot, and J. Frey), Rep. No. EUR 19245 EN, European Commission, Luxembourg, 109-112,
  • 8. Gonçalves R, Regalla J, Nicolet J, Frey J, Nicholas R, Bashiruddin J, De Santis P, Gonçalves AP (1998). Antigen heterogeneity among Mycoplasma mycoides subsp. mycoides SC isolates: discrimination of major surface proteins. Veterinary Microbiology, 63(1):13-28.
  • 9. Erdağ O, Türkaslan J (1994). Sığırların süreğen solunum yolu hastalığı (CBPP)’nin serolojik testler ile araştırılması. Pendik Vet Mikrobiyol Derg. 25(1-2): 49-53.
  • 10. Çetinkaya B, Öngör H, Karahan M, Kalender H, Lorenzon S, Thiaucourt F (2003). Abattoir-based survey of contagious bovine pleuropneumonia in cattle in Turkey, Veterinary Record.152:254-258.
  • 11. .Miles, R. J., & Nicholas, R. (1998). Mycoplasma Protocols (Methods in Molecular Biology, vol. 104).
  • 12. . Noordhuizen, JPTM, Frankena, K, Thrusfield, MV, Graat, EAM (1997). Application of quantitative methods in veterinary epidemiology. Wageningen Pers Publ. 2nd ed. 2001, Wageningen The Netherlands, 429.coplasma protocols Ed. R.A.J Miles and R.A.J Nicholas, Humana Pres Inc. 37-51.
  • 13. Duffy, MF., Noormohammadi, AH., Baseggio, N., Browning, GF. and Markham, PF (1998). Polyacrylamide gel-electrophoresis separation of whole-cell proteins. Methods in molecular biology, Vol 104: Mycoplasma protocols Ed. R.A.J Miles and R.A.J Nicholas, Humana Pres Inc., Totowa, NJ, 267-277.
  • 14. Towbin, S.T., Staehelin, T. and Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrilamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Academy of Science, USA, 76, 4350-4354.
  • 15. Amanfu, W., Sediadie S., Masupu K.V., Benkirane A., Geiger R. & Thiaucourt F. (1998). Field validation of a competitive ELISA for the detection of contagious bovine pleuropneumonia in Botswana. Rev. Elev. Med. Vet. Pays Trop. 51:189–193.
  • 16. . Ayling, R, Miles, RJ., Nicholas, RAJ. (1999). Development of improved serological tests for the diagnosis of CBPP. Proceedings of the International Symposium on Mycoplasmas of Ruminants. Toulouse, France, p.35.
  • 17. Yansambou MS, Ferreira AC, Bada-Alambedji R, Botelho A (2018) Seroprevalence of Contagious Bovine Pleuropneumonia in Niamey Region, Niger. J Vet Med Res 5(8): 1151.
  • 18. Regalla, J., Gonçalves, R., Niza Ribeiro, J., Duarte, L., Nicholas, R., Bashiruddin, J., De Santis, P., Garrido Abellan, F. and Penha Gonçalves, A. (1999). Development of immunoblotting as a diagnostic tool for contagious bovine pleuropneumonia. In COST 826 Agriculture and biotechnology “Mycoplasmas of ruminants: pathogenicity, diagnostics, epidemiology and molecular genetics”, (Ed. D. Bergonier, X. Berthelot, and J. Frey), Rep. No. EUR 19245 EN, European Commission, Luxembourg, 109-112.
  • 19. Abdo, E. M., Nicolet, J., Miserez, R., Goncalves, R., Regalla, J., Griot, C., Bensaide, A., Krampe, M. & Frey, J. (1998). Humoral and bronchial immune responses in cattle experimentally infected with Mycoplasma mycoides subsp. mycoides small colony type, Vet Microbiol, 59, 109–122.
  • 20. Cheng, X., J. Nicolet, R. Miserez, P. Kuhnert, M. Krampe, T. Pilloud, E. M. Abdo, C. Griot, and J. Frey. (1996). Characterization of the gene for an immunodominant 72 kDa lipoprotein of Mycoplasma mycoides subsp. mycoides small colony type, Microbiology, 142,3515-3524
  • 21. Le Goff, C. & Thiaucourt, F. (1998). A competitive ELISA for the specific diagnosis of contagious bovine pleuropneumonia (CBPP), Vet Microbiol, 60, 179–191.

The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle with Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-bent Assay and Complement Fixation Test

Year 2021, , 113 - 117, 31.12.2021
https://doi.org/10.47027/duvetfd.995044

Abstract

In this study 253 serum samples from cattle showing respiratory disorders were tested by confirmatory Western Immunoblotting test (IBT), complement fixation test (CFT) and competitive enzyme linked immunosorbent assay (c-ELISA). Two (0.8%) out of 253 serum samples were found to be positive and two were (0.8%) doubtful by CFT while 7 (2.8%) serum samples were found as positive and 25 (9.9%) of them were found to be doubtful by c-ELISA. On the other hand, a core profile of antigenic bands needed to be identified at 110, 98, 95, 62/60 and 48 kDa was not detected in any of the test serum sample. Immunoblot analysis some of serum samples displayed the number of bands between one or three and some of them were highly faint. In order to detect similarities between MmmSc strains related to their electrophoretic profiles, whole cell proteins of Mycoplasma species isolated from cattle and MmmSC reference strains (Mycoplasma mycoides subsp. mycoides SC Botswana (African strain), Mycoplasma mycoides subsp. mycoides SC PG1 (reference strain), Mycoplasma mycoides subsp. mycoides SC V5 (Australian strain), M. bovigenitalium, M. bovirhinis, M. capricolum subsp. capricolum, M.canadense, M.alkalescens and M. bovis) were analysed by sodium dodecyl sulphate polyacrilamide gel elektrophoresis (SDS-PAGE). The result of this analysis indicated that antigens of 220, 78, 82, 68, 24 kDa were common among all tested strains. According to these results, it was thought that positive results by both CFT and c-ELISA, also non specific band patterns observed by IBT might be caused by a cross reaction between tested mycoplasma strains. As conclusion, it is recommended that all the positive results by CFT and c-ELISA be confirmed by IBT.

References

  • 1. Nicholas RA, Bashiruddin JB, Ayling RD, Miles RJ (2000). Contagious bovine pleuropneumonia: a review of recent developments. Vet Bull. 70:827-838.
  • 2. OIE. (2015). Contagious bovine pleuropneumonia. In: Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Parisi
  • 3. Wawegama NK., & Browning GF (2017). Improvements in diagnosis of disease caused by Mycoplasma bovis in cattle. Animal Production Science. 57(7): 1482-1487.
  • 4. Amanfu W. (2019) Contagious Bovine Pleuropneumonia. In: Kardjadj M., Diallo A., Lancelot R. (eds) Transboundary Animal Diseases in Sahelian Africa and Connected Regions. Springer, Cham.
  • 5. Nicholas RAJ, Santini FG, Clark KM, Palmer NM, Santis P De, Bashiruddin JB (1996). A comparison of serological tests and gross lung pathology for detecting contagious bovine pleuropneumonia in two groups of Italian cattle, Vet Rec. 139: 89-93,
  • 6. Regalla J, Gonçalves R, Ribeiro N, Duarte L, Penha Gonçalves A (1996). Use of immunoblotting test for carrier state definition in contagious bovine pleuropneumonia, In Mycoplasmas of ruminants: pathogenicity, diagnostics, epidemiology and molecular genetics, Ed. J. Frey and K . Sarris, Rep, No. EUR 16934 EN, European Commission, Luxembourg, 72-74.
  • 7. Regalla J, Gonçalves R, Niza Ribeiro J, Duarte L, Nicholas R, Bashiruddin J, De Santis P, Garrido Abellan F, Penha Gonçalves A (1999). Development of immunoblotting as a diagnostic tool for contagious bovine pleuropneumonia. In, COST 826 Agriculture and biotechnology “Mycoplasmas of ruminants: pathogenicity, diagnostics, epidemiology and molecular genetics”, (Ed. D. Bergonier, X. Berthelot, and J. Frey), Rep. No. EUR 19245 EN, European Commission, Luxembourg, 109-112,
  • 8. Gonçalves R, Regalla J, Nicolet J, Frey J, Nicholas R, Bashiruddin J, De Santis P, Gonçalves AP (1998). Antigen heterogeneity among Mycoplasma mycoides subsp. mycoides SC isolates: discrimination of major surface proteins. Veterinary Microbiology, 63(1):13-28.
  • 9. Erdağ O, Türkaslan J (1994). Sığırların süreğen solunum yolu hastalığı (CBPP)’nin serolojik testler ile araştırılması. Pendik Vet Mikrobiyol Derg. 25(1-2): 49-53.
  • 10. Çetinkaya B, Öngör H, Karahan M, Kalender H, Lorenzon S, Thiaucourt F (2003). Abattoir-based survey of contagious bovine pleuropneumonia in cattle in Turkey, Veterinary Record.152:254-258.
  • 11. .Miles, R. J., & Nicholas, R. (1998). Mycoplasma Protocols (Methods in Molecular Biology, vol. 104).
  • 12. . Noordhuizen, JPTM, Frankena, K, Thrusfield, MV, Graat, EAM (1997). Application of quantitative methods in veterinary epidemiology. Wageningen Pers Publ. 2nd ed. 2001, Wageningen The Netherlands, 429.coplasma protocols Ed. R.A.J Miles and R.A.J Nicholas, Humana Pres Inc. 37-51.
  • 13. Duffy, MF., Noormohammadi, AH., Baseggio, N., Browning, GF. and Markham, PF (1998). Polyacrylamide gel-electrophoresis separation of whole-cell proteins. Methods in molecular biology, Vol 104: Mycoplasma protocols Ed. R.A.J Miles and R.A.J Nicholas, Humana Pres Inc., Totowa, NJ, 267-277.
  • 14. Towbin, S.T., Staehelin, T. and Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrilamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Academy of Science, USA, 76, 4350-4354.
  • 15. Amanfu, W., Sediadie S., Masupu K.V., Benkirane A., Geiger R. & Thiaucourt F. (1998). Field validation of a competitive ELISA for the detection of contagious bovine pleuropneumonia in Botswana. Rev. Elev. Med. Vet. Pays Trop. 51:189–193.
  • 16. . Ayling, R, Miles, RJ., Nicholas, RAJ. (1999). Development of improved serological tests for the diagnosis of CBPP. Proceedings of the International Symposium on Mycoplasmas of Ruminants. Toulouse, France, p.35.
  • 17. Yansambou MS, Ferreira AC, Bada-Alambedji R, Botelho A (2018) Seroprevalence of Contagious Bovine Pleuropneumonia in Niamey Region, Niger. J Vet Med Res 5(8): 1151.
  • 18. Regalla, J., Gonçalves, R., Niza Ribeiro, J., Duarte, L., Nicholas, R., Bashiruddin, J., De Santis, P., Garrido Abellan, F. and Penha Gonçalves, A. (1999). Development of immunoblotting as a diagnostic tool for contagious bovine pleuropneumonia. In COST 826 Agriculture and biotechnology “Mycoplasmas of ruminants: pathogenicity, diagnostics, epidemiology and molecular genetics”, (Ed. D. Bergonier, X. Berthelot, and J. Frey), Rep. No. EUR 19245 EN, European Commission, Luxembourg, 109-112.
  • 19. Abdo, E. M., Nicolet, J., Miserez, R., Goncalves, R., Regalla, J., Griot, C., Bensaide, A., Krampe, M. & Frey, J. (1998). Humoral and bronchial immune responses in cattle experimentally infected with Mycoplasma mycoides subsp. mycoides small colony type, Vet Microbiol, 59, 109–122.
  • 20. Cheng, X., J. Nicolet, R. Miserez, P. Kuhnert, M. Krampe, T. Pilloud, E. M. Abdo, C. Griot, and J. Frey. (1996). Characterization of the gene for an immunodominant 72 kDa lipoprotein of Mycoplasma mycoides subsp. mycoides small colony type, Microbiology, 142,3515-3524
  • 21. Le Goff, C. & Thiaucourt, F. (1998). A competitive ELISA for the specific diagnosis of contagious bovine pleuropneumonia (CBPP), Vet Microbiol, 60, 179–191.
There are 21 citations in total.

Details

Primary Language English
Subjects Veterinary Surgery
Journal Section Research
Authors

Sevil Erdenliğ Gürbilek 0000-0002-0377-2650

Ümit Özdemir

Osman Yaşar Tel 0000-0001-7848-3899

Publication Date December 31, 2021
Acceptance Date October 15, 2021
Published in Issue Year 2021

Cite

APA Erdenliğ Gürbilek, S., Özdemir, Ü., & Tel, O. Y. (2021). The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle with Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-bent Assay and Complement Fixation Test. Dicle Üniversitesi Veteriner Fakültesi Dergisi, 14(2), 113-117. https://doi.org/10.47027/duvetfd.995044
AMA Erdenliğ Gürbilek S, Özdemir Ü, Tel OY. The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle with Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-bent Assay and Complement Fixation Test. Dicle Üniv Vet Fak Derg. December 2021;14(2):113-117. doi:10.47027/duvetfd.995044
Chicago Erdenliğ Gürbilek, Sevil, Ümit Özdemir, and Osman Yaşar Tel. “The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle With Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-Bent Assay and Complement Fixation Test”. Dicle Üniversitesi Veteriner Fakültesi Dergisi 14, no. 2 (December 2021): 113-17. https://doi.org/10.47027/duvetfd.995044.
EndNote Erdenliğ Gürbilek S, Özdemir Ü, Tel OY (December 1, 2021) The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle with Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-bent Assay and Complement Fixation Test. Dicle Üniversitesi Veteriner Fakültesi Dergisi 14 2 113–117.
IEEE S. Erdenliğ Gürbilek, Ü. Özdemir, and O. Y. Tel, “The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle with Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-bent Assay and Complement Fixation Test”, Dicle Üniv Vet Fak Derg, vol. 14, no. 2, pp. 113–117, 2021, doi: 10.47027/duvetfd.995044.
ISNAD Erdenliğ Gürbilek, Sevil et al. “The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle With Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-Bent Assay and Complement Fixation Test”. Dicle Üniversitesi Veteriner Fakültesi Dergisi 14/2 (December 2021), 113-117. https://doi.org/10.47027/duvetfd.995044.
JAMA Erdenliğ Gürbilek S, Özdemir Ü, Tel OY. The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle with Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-bent Assay and Complement Fixation Test. Dicle Üniv Vet Fak Derg. 2021;14:113–117.
MLA Erdenliğ Gürbilek, Sevil et al. “The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle With Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-Bent Assay and Complement Fixation Test”. Dicle Üniversitesi Veteriner Fakültesi Dergisi, vol. 14, no. 2, 2021, pp. 113-7, doi:10.47027/duvetfd.995044.
Vancouver Erdenliğ Gürbilek S, Özdemir Ü, Tel OY. The Evaluation of Humoral Immune Response to Contagious Bovine Pleuropneumonia in Cattle with Respiratory Disorders by Western Blot Technique, Competitive Enzyme Linked Immunosor-bent Assay and Complement Fixation Test. Dicle Üniv Vet Fak Derg. 2021;14(2):113-7.