Comparative evaluation of outer membrane protein and whole cell antigen vaccine against avian pathogen Escherichia coli infection in broiler chicken
-
Basharat Maqbool Wani
,
Shayaib Ahmad Kamil
,
Showkat Ahmad Shah
,
Masood Saleem Mir
,
Mir Nadeem Hassan
,
Zahid Amin Kashoo
,
Mudasir Ali Rather
,
Majid Shafi
,
Zahoor Ahmad Wani
,
Nadeem Shabir
,
Nazli Tizro
,
Akeel Bashir
,
Atif Khurshid Wani
Abstract
Background and Objectives: Present study was aimed at assessing protective efficacy of outer membrane protein (OMP) vaccine in comparison to inactivated whole cell antigen vaccine after challenge with homologous serogroup (O2) of avian pathogenic Escherichia coli in broiler chickens.
Materials and Methods: The outer membrane proteins were extracted by sarcosyl method and protein concentration was determined by nanodrop spectrophotometer. The study comprised of 120 birds divided into 6 groups. The birds were subcutaneously immunized twice with primary vaccine in the first week followed by booster vaccine in second week.
Results: The protection rate of 82% was found in whole cell inactivated antigen, 91% in OMP vaccine and 27% among the unvaccinated group. The antibody (IgG) response was found significantly higher in OMP vaccine group than whole cell antigen group. In unvaccinated groups chicks, the antibody titer never reached to the protective level till the termination of experiment. The bacteria were re-isolated from the infected broiler chickens for the confirmation of induced infection and were characterized using standard cultural and biochemical tests belonging to O2 serogroup.
Conclusion: Our study demonstrated that the outer membrane protein (OMP) vaccine provided significantly higher protection (91%) and antibody response compared to the inactivated whole cell antigen vaccine (82%) against Escherichia coli O2 infection in broiler chickens. Birds vaccinated with OMP exhibited fewer pathological lesions and a stronger immune response. The findings underscore the potential of OMP-based vaccines as a safer, more immunogenic alternative for controlling colibacillosis.
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2. Amin U, Kamil SA, Wani BM, Qureshi S, Shah SA, Dar TA, et al. Haematological and biochemical alterations of broiler chicken affected naturally with colibacillosis. Int J Curr Microbiol App Sci 2020; 9: 1906-1913.
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7. Li L, Thofner I, Christensen JP, Ronco T, Pedersen K, Olsen RH. Evaluation of the efficacy of an autogenous Escherichia coli vaccine in broiler breeders. Avian Pathol 2017; 46: 300-308.
8. Ghunaim H, Abu-Madi MA, Kariyawasam S. Advances in vaccination against avian pathogenic Escherichia coli respiratory disease: Potentials and limitations. Vet Microbiol 2014; 172: 13-22.
9. Zhang XL, Yang F, Zou JT, Wu WR, Jing HM, Gou Q, et al. Immunization with Pseudomonas aeruginosa outer membrane vesicles stimulates protective immunity in mice. Vaccine 2018; 36: 1047-1054.
10. Liu Q, Liu Q, Yi J, Liang K, Liu T, Roland KL, et al. Outer membrane vesicles derived from Salmonella Typhimurium mutants with truncated LPS induce cross-protective immune responses against infection of Salmonella enterica serovars in the mouse model. Int J Med Microbiol 2016; 306: 697-706.
11. Koebnik R, Locher KP, Van Gelder P. Structure and function of bacterial outer membrane proteins: barrels in a nutshell. Mol Microbiol 2000; 37: 239-253.
12. Osman KM, Marouf SH. Comparative dendroGram analysis of OMPs of Salmonella enteric serotype Enteritidis with Typhimurium, Braendurup and Lomita isolated from pigeons. Int J Adv Biotechnol Res 2014; 2:952-960.
13. Gao S, Peng DX, Wen QY, Jiao XA, Gan, JJ, Wu YT, et al. Study on the Protection Mechanism of the O18 and O78 Isolates Based on Ultrasonic Inactivation Vaccine of Avian Escherichia coli, J. (in Chinese). Chin J Anim Vet Sci 2001 32: 345-353.
14. Liu C, Chen Z, Tan C, Liu W, Xu Z, Zhou R, et al. Immunogenic characterization of outer membrane porins OmpC and OmpF of porcine extraintestinal pathogenic Escherichia coli. FEMS Microbiology Letters 2012; 337: 104-111.
15. Tabatabai LB, Zehr ES. Identification of five outer membrane-associated proteins among cross-protective factor proteins of Pasteurella multocida. Infect Immun 2004;72:1195-1198.
16. Sambrook J, Russell DW (2001). Molecular Cloning: A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.
17. Arthur SA, Singh SD, Dhama K. Efficacy of outer membrane protein vaccine against avian pathogenic E. coli in broiler chicks. IJPS 2010; 45: 186-190.
18. Acevedo-Villanueva K, Akerele G, Al-Hakeem W, Adams D, Gourapura R, Selvaraj R. Immunization of Broiler Chickens with a killed Chitosan nanoparticle Salmonella vaccine decreases Salmonella Enterica serovar Enteritidis load. Front Physiol 2022;13:920777.
19. Boone, D. R., Castenholz, R. W., & Garrity, G. M. Bergey's manual of systematic bacteriology (2nd ed.). Springer. 2001.
20. Zapata A, Ramirez-Arcos S. A comparative study of McFarland turbidity standards and the Densimat photometer to determine bacterial cell density. Curr Microbiol 2015; 70:907-909.
21. El Jakee JK, El Amry GM, Hessain AM, Hemeg HA, Shafei SM, Moussa IM. Production and evaluation of autogenous vaccine against avian colibacillosis. J Anim Plant Sci 2016; 26: 79-87.
22. Gao D, Yu J, Dai X, Tian Y, Sun J, Xu X, Cai X. Development and evaluation of an indirect enzymelinked immunosorbent assay based on a recombinant SifA protein to detect Salmonella infection in poultry. Poult Sci 2023; 102:102513.
23. Mishra A, Shard R, Chhabra D, Moghe MN. E. coli isolates from domestic poultry. Indian J Anim Sci 2002; 72: 727-729.
24. Shah SA, Mir MS, Kamil SA, Shafi M, Adil S,Wani BM, et al. Prevalence and isolation of avian pathogenic Escherichia coli from colibacillosis affected broiler chicken in Kashmir valley. Life Sci Leafl 2020; 125: 06-13.
25. Mitra S, Barman S, Nag D, Sinha R, Saha DR, Koley H. Outer membrane vesicles of Shigella boydii type 4 induce passive immunity in neonatal mice. FEMS Immunol Med Microbiol 2012; 66: 240-250.
26. Shivachandra SB, Kumar A, Yogisharadhya R, Viswas KN. Immunogenicity of highly conserved recombinant VacJ outer membrane lipoprotein of Pasteurella multocida. Vaccine 2014;32:290-296.
27. Chen HY, Huang QY, Chen GZ. Immunogenicity of the outer membrane proteins of poultry-sourced E. coli O2 and O78, their antibiotic-resistant, and fusion strains [J] (in Chinese). J South China Agric Univ 2005; 26: 106-109.
28. Vaef Zadeh F, Esmaily F, Sharifi-Yazdi MK. Protective Immune Responses Induced in Chickens by Outer Membrane Proteins Extracted from Different Strains of Escherichia coli. Iran J Allergy Asthma Immunol 2004; 3: 133-137.
29. Tie Z, Guang CW, Xinghua Z. Clinical isolating outer membrane protein pattern from Avian Escherichia coli of China. Afr J Microbiol Res 2010; 4: 23-30.
30. Chen Y, Jie K, Li B, Yu H, Ruan H, Wu J, Huang X, Liu Q. Immunization with Outer Membrane Vesicles Derived from Major Outer Membrane Protein-Deficient Salmonella Typhimurium Mutants for Cross Protection Against Salmonella Enteritidis and Avian Pathogenic Escherichia coli O78 Infection in Chickens. Front Microbiol 2020; 11:588952.
31. Kumar A, Bhalerao D, Gupta RP, Kumari M. Pathological Studies on Natural Cases of Colibacillosis in Haryana State. Haryana Vet 2013; 52: 118-120.
32. Wani BM, Kamil SA, Shah SA, Shafi M, Shabir M, Kashani B, et al. Isolation and biochemical characterization of avian pathogenic Escherichia coli from different organs in colibacillosis affected broiler chicken. J Entomol Zool Stud 2020 ;8 :1649-1652.
33. Renu AK, Mishra SK, Londhe MS, Deepika L, Anshu S. Etio-Pathological studies on poultry mortality with reference to Escherichia coli infections. Indian J Poult Sci 2012; 47: 222-226.
34. Gangane GR, Kulkarni GB, Yeotikar PV. Studies on experimental Colibacillosis in chicks. Indian Vet J 2006; 83: 118-119.
35. Sylvester SA, Singh SD, Mahender M. In-ovo and In-vivo pathogenicity study of avian Escherichia coli isolated from cases of colibacillosis in chickens. Indian J Vet Pathol 2005; 3: 225-230.
36. Shah SA, Mir MS, Wani BM, Kamil SA, Goswami P, Amin U, et al. Pathological studies on avian pathogenic Escherichia coli infection in broilers. Pharm Innov J 2019; 8: 68-73.
37. Hegazy AM, Abd-ElSamie LK, ElSayed EM. The immunosuppressive effect of E. coli in chickens vaccinated with infectious bronchitis (IB) or infectious bursal disease (IBD) vaccines. J Am Sci 2010; 6: 762-767.
38. Ashish Hooda A, Mishra SK, Vikas Nehra VN, Deepika Lather DL. Patho-Anatomical Studies on Poultry with Special Reference to Gastro-Intestinal tract disorders. Haryana Vet 2011; 50: 80-84.
2. Amin U, Kamil SA, Wani BM, Qureshi S, Shah SA, Dar TA, et al. Haematological and biochemical alterations of broiler chicken affected naturally with colibacillosis. Int J Curr Microbiol App Sci 2020; 9: 1906-1913.
3. Wang SQ, Peng, HM, Jia XF, Zeng JL, Zhu CL, et al. Prevention of Escherichia coli infection in broiler chickens with Lactobacillus plantarum B. Poult Sci 2017; 96:2576-2586.
4. Johnson TJ, Wannemuehler Y, Johnson SJ, Stell AL, Doetkott C, Johnson JR, et al. Comparison of extraintestinal pathogenic Escherichia coli strains from human and avian sources reveals a mixed subset representing potential zoonotic pathogens. Appl Environ Microb 2008; 74: 7043-7050.
5. Bélanger L, Garenaux A, Harel J, Boulianne M, Nadeau E, Dozois CM. Escherichia coli from animal reservoirs as a potential source of human extraintestinal pathogenic E. coli. FEMS Immunol Med Mic 2011; 62: 1-10.
6. Lu Q, Zhang W, Luo L, Wang H, Shao H, Zhang T, et al. Genetic diversity and multidrug resistance of phylogenic groups B2 and D in InPEC and ExPEC isolated from chickens in Central China. BMC Microbiol 2022; 22: 60.
7. Li L, Thofner I, Christensen JP, Ronco T, Pedersen K, Olsen RH. Evaluation of the efficacy of an autogenous Escherichia coli vaccine in broiler breeders. Avian Pathol 2017; 46: 300-308.
8. Ghunaim H, Abu-Madi MA, Kariyawasam S. Advances in vaccination against avian pathogenic Escherichia coli respiratory disease: Potentials and limitations. Vet Microbiol 2014; 172: 13-22.
9. Zhang XL, Yang F, Zou JT, Wu WR, Jing HM, Gou Q, et al. Immunization with Pseudomonas aeruginosa outer membrane vesicles stimulates protective immunity in mice. Vaccine 2018; 36: 1047-1054.
10. Liu Q, Liu Q, Yi J, Liang K, Liu T, Roland KL, et al. Outer membrane vesicles derived from Salmonella Typhimurium mutants with truncated LPS induce cross-protective immune responses against infection of Salmonella enterica serovars in the mouse model. Int J Med Microbiol 2016; 306: 697-706.
11. Koebnik R, Locher KP, Van Gelder P. Structure and function of bacterial outer membrane proteins: barrels in a nutshell. Mol Microbiol 2000; 37: 239-253.
12. Osman KM, Marouf SH. Comparative dendroGram analysis of OMPs of Salmonella enteric serotype Enteritidis with Typhimurium, Braendurup and Lomita isolated from pigeons. Int J Adv Biotechnol Res 2014; 2:952-960.
13. Gao S, Peng DX, Wen QY, Jiao XA, Gan, JJ, Wu YT, et al. Study on the Protection Mechanism of the O18 and O78 Isolates Based on Ultrasonic Inactivation Vaccine of Avian Escherichia coli, J. (in Chinese). Chin J Anim Vet Sci 2001 32: 345-353.
14. Liu C, Chen Z, Tan C, Liu W, Xu Z, Zhou R, et al. Immunogenic characterization of outer membrane porins OmpC and OmpF of porcine extraintestinal pathogenic Escherichia coli. FEMS Microbiology Letters 2012; 337: 104-111.
15. Tabatabai LB, Zehr ES. Identification of five outer membrane-associated proteins among cross-protective factor proteins of Pasteurella multocida. Infect Immun 2004;72:1195-1198.
16. Sambrook J, Russell DW (2001). Molecular Cloning: A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.
17. Arthur SA, Singh SD, Dhama K. Efficacy of outer membrane protein vaccine against avian pathogenic E. coli in broiler chicks. IJPS 2010; 45: 186-190.
18. Acevedo-Villanueva K, Akerele G, Al-Hakeem W, Adams D, Gourapura R, Selvaraj R. Immunization of Broiler Chickens with a killed Chitosan nanoparticle Salmonella vaccine decreases Salmonella Enterica serovar Enteritidis load. Front Physiol 2022;13:920777.
19. Boone, D. R., Castenholz, R. W., & Garrity, G. M. Bergey's manual of systematic bacteriology (2nd ed.). Springer. 2001.
20. Zapata A, Ramirez-Arcos S. A comparative study of McFarland turbidity standards and the Densimat photometer to determine bacterial cell density. Curr Microbiol 2015; 70:907-909.
21. El Jakee JK, El Amry GM, Hessain AM, Hemeg HA, Shafei SM, Moussa IM. Production and evaluation of autogenous vaccine against avian colibacillosis. J Anim Plant Sci 2016; 26: 79-87.
22. Gao D, Yu J, Dai X, Tian Y, Sun J, Xu X, Cai X. Development and evaluation of an indirect enzymelinked immunosorbent assay based on a recombinant SifA protein to detect Salmonella infection in poultry. Poult Sci 2023; 102:102513.
23. Mishra A, Shard R, Chhabra D, Moghe MN. E. coli isolates from domestic poultry. Indian J Anim Sci 2002; 72: 727-729.
24. Shah SA, Mir MS, Kamil SA, Shafi M, Adil S,Wani BM, et al. Prevalence and isolation of avian pathogenic Escherichia coli from colibacillosis affected broiler chicken in Kashmir valley. Life Sci Leafl 2020; 125: 06-13.
25. Mitra S, Barman S, Nag D, Sinha R, Saha DR, Koley H. Outer membrane vesicles of Shigella boydii type 4 induce passive immunity in neonatal mice. FEMS Immunol Med Microbiol 2012; 66: 240-250.
26. Shivachandra SB, Kumar A, Yogisharadhya R, Viswas KN. Immunogenicity of highly conserved recombinant VacJ outer membrane lipoprotein of Pasteurella multocida. Vaccine 2014;32:290-296.
27. Chen HY, Huang QY, Chen GZ. Immunogenicity of the outer membrane proteins of poultry-sourced E. coli O2 and O78, their antibiotic-resistant, and fusion strains [J] (in Chinese). J South China Agric Univ 2005; 26: 106-109.
28. Vaef Zadeh F, Esmaily F, Sharifi-Yazdi MK. Protective Immune Responses Induced in Chickens by Outer Membrane Proteins Extracted from Different Strains of Escherichia coli. Iran J Allergy Asthma Immunol 2004; 3: 133-137.
29. Tie Z, Guang CW, Xinghua Z. Clinical isolating outer membrane protein pattern from Avian Escherichia coli of China. Afr J Microbiol Res 2010; 4: 23-30.
30. Chen Y, Jie K, Li B, Yu H, Ruan H, Wu J, Huang X, Liu Q. Immunization with Outer Membrane Vesicles Derived from Major Outer Membrane Protein-Deficient Salmonella Typhimurium Mutants for Cross Protection Against Salmonella Enteritidis and Avian Pathogenic Escherichia coli O78 Infection in Chickens. Front Microbiol 2020; 11:588952.
31. Kumar A, Bhalerao D, Gupta RP, Kumari M. Pathological Studies on Natural Cases of Colibacillosis in Haryana State. Haryana Vet 2013; 52: 118-120.
32. Wani BM, Kamil SA, Shah SA, Shafi M, Shabir M, Kashani B, et al. Isolation and biochemical characterization of avian pathogenic Escherichia coli from different organs in colibacillosis affected broiler chicken. J Entomol Zool Stud 2020 ;8 :1649-1652.
33. Renu AK, Mishra SK, Londhe MS, Deepika L, Anshu S. Etio-Pathological studies on poultry mortality with reference to Escherichia coli infections. Indian J Poult Sci 2012; 47: 222-226.
34. Gangane GR, Kulkarni GB, Yeotikar PV. Studies on experimental Colibacillosis in chicks. Indian Vet J 2006; 83: 118-119.
35. Sylvester SA, Singh SD, Mahender M. In-ovo and In-vivo pathogenicity study of avian Escherichia coli isolated from cases of colibacillosis in chickens. Indian J Vet Pathol 2005; 3: 225-230.
36. Shah SA, Mir MS, Wani BM, Kamil SA, Goswami P, Amin U, et al. Pathological studies on avian pathogenic Escherichia coli infection in broilers. Pharm Innov J 2019; 8: 68-73.
37. Hegazy AM, Abd-ElSamie LK, ElSayed EM. The immunosuppressive effect of E. coli in chickens vaccinated with infectious bronchitis (IB) or infectious bursal disease (IBD) vaccines. J Am Sci 2010; 6: 762-767.
38. Ashish Hooda A, Mishra SK, Vikas Nehra VN, Deepika Lather DL. Patho-Anatomical Studies on Poultry with Special Reference to Gastro-Intestinal tract disorders. Haryana Vet 2011; 50: 80-84.
| Files | ||
| Issue | Vol 18 No 1 (2026) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v18i1.20935 | |
| Keywords | ||
| Escherichia coli Colibacillosis Chickens Bacterial outer membrane proteins Vaccines Immune response | ||
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How to Cite
1.
Wani B, Ahmad Kamil S, Shah S, Saleem Mir M, Hassan MN, Amin Kashoo Z, Rather MA, Shafi M, Ahmad Wani Z, Shabir N, Tizro N, Bashir A, Wani A. Comparative evaluation of outer membrane protein and whole cell antigen vaccine against avian pathogen Escherichia coli infection in broiler chicken. Iran J Microbiol. 2026;18(1):91-101.
