Isolation and characterization of outer membrane proteins (OMPs) from Salmonella Gallinarum in chicken and antibiogram of the isolates
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Abstract
Salmonella isolates should be distinguished as it may assist in tracing the source of an outbreak and monitoring trends in antimicrobial resistance associated with a particular type. The specific detection of these Salmonella serotypes is therefore extremely important in order to attribute an isolate to a previously known epidemic outbreak. The present investigation was to isolate and identify S. Gallinarum, to study variation in the profile of outer membrane proteins (OMPs) and to determine in vitro antibiogram of S. Gallinarum in poultry. A total of 228 faecal samples and 22 visceral samples suspected for Salmonellosis were collected, of these 15 samples (6.0%) were found positive for S. Gallinarum. In the present study, rfbS gene sequence was helpful in the serotype-specific detection of S. Gallinarum giving a 187 bp product. Salmonella Gallinarum crude protein extracts determined by SDSPAGE showed migration of OMPs as several bands at approximate moleculer weights of appx. 45 kDa, 55 kDa, 64 kDa, 65 kDa, 74 kDa, 110 kDa, 120 kDa, 135 kDa, 150 kDa,155 kDa, 200 kDa and above 200 kDa. The study indicated a definite variation in the profile of OMPs of various Salmonella Gallinarum strains with major OMPs in the range of appx 80-100 kDa which could be the target for vaccine production. All the isolates tested against 14 antimicrobial agents showed variable susceptibility pattern with highest resistance to nalidixic acid, ampicillin and sulphadiazine and sensitivity to chloramphenicol, gentamicin and enrofloxacin.
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Keywords
Antibiogram, Outer membrane protein (OMP), PCR, Salmonella Gallinarum, SDS-PAGE
References
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Bauer, A.W., Bauer, Kirby, W.M., Kirby, Sherris, J.C., Sher-ris and Turck , M., Turck. (1966). Antibiotic suscepti-bility testing by a standardized single disk method. American Journal of Clinical Pathology, 45(4): 493-496
Bouzoubaa, K. Bouzoubaa, Nagaraja, K.V. Nagaraja, Kab-baj, F.Z. Kabbaj, Newman, J.A. Newman and Pomeroy, B.S. Pomeroy (1989). Feasibility of using proteins from Salmonella Gallinarum vs. 9R live vaccine for the pre-vention of fowl typhoid in chickens. Avian. Dis., 33: 385-391
Bouzoubaa, K. Bouzoubaa, Nagaraja, K.V. Nagaraja, New-man, J.A. Newman and Pomeroy, B.S. Pomeroy (1987). Use of membrane proteins from Salmonella Gallinarum for prevention of fowl typhoid infection in chickens. Avian. Dis., 31: 699-704
Carraminana, J.J. Carraminana, Rota, C. Rota, Agustín, I. Agustín and Herrera, A. Herrera (2004). High preva-lence of multiple resistance to antibiotics in Salmonella serovars isolated from a poultry slaughterhouse in Spain. Veterinary Microbiology, 104: 133-139
Choi-Kim, K. Choi-Kim, Maheswaran, S.K. Maheswaran, Felice, L.J. Felice and Molitor, T.W. Molitor (1991). Relationship between the iron regulated outer mem-brane proteins and the outer membrane proteins of in vivo grown Pasteurella multocida. Veterinary Microbi-ology, 28(1): 75-92
Christensen, J.P. Christensen, Olsen, J.E. Olsen and Bis-gaard, M. Bisgaard. (1992a). Characterization of Salmo-nella enterica serotype gallinarum biotypes gallinarum and pullorum by plasmid profiling and biochemical analysis. Avian Pathology, 21: 461 -470
Christensen, J.P. Christensen, Olsen, J.E. Olsen and Bis-gaard, M. Bisgaard (1992b). Ribotypes of Salmonella enterica serovar Gallinarum biovars Gallinarum and Pullorum. Avian Pathology, 22: 725–738
Davies, R.H. Davies and Breslin, M. Breslin (2003). Obser-vations on Salmonella contamination of commercial laying farms before and after cleaning and disinfection. Veterinary Records, 152: 283-287
Desai, A.R., Shah, D.H., Shringi, S., Lee, M.J., Li, Y. H., Cho, M.R., Park, J.H., Eo, S.K, Lee, J.H,, Chae, J. S. (2005). An allele-specific PCR assay for the rapid and serotype-specific detection of Salmonella Pullorum. Avian Dis., 49(4): 558-61
Gast, R.K. Gast (1997). Salmonella infections in Diseases of Poultry. Iowa State University Press, Ames, pp. 81-121.
Holt, J.G Holt, Krieg, N.R. Krieg, Sneath, P.H.A. Sneath and Williams, S.T. Williams. (1994). Bergey’s manual of determinative bacteriology, 9th Ed. Lippincott, Wil-liams & Wilkins, Baltimore, Maryland.
Laemmli, U.K. Laemmli. (1970). Cleavage of structural proteins during the assembly of the head of bacterio-phage T4. Nature, 227(5259): 680–685
Lee, Y.J. Lee, Kim, K.S. Kim, Kwon, Y.K. Kwon and Tak, R.B. Tak. (2003). Biochemical characteristics and anti-microbials susceptibility of Salmonella Gallinarum isolated in Korea. Journal of Veterinary Sciences, 4(2): 161-166
Lee, Y.S. Lee, Chol, K.S. Chol, Kim, M.C. Kim, Han, J.C. Han, Park, J.H. Park, Shah, D.H. Shah and Chae, J.S. Chae. (2005). Purification of outer membrane protein (OMP C) from Salmonella Gallinarum for characteriza-tion of its immunogenicity. Indian Journal of Animal Sciences, 75(2): 164-167
Liu, D., Liu, Verma N.K., Verma, Romana, L.K. Ro,mana and Reeves, P.R. Reeves. (1991). Completing the se-quence of the rfb region of Salmonella serovars typhi (group D) and paratyphi (group A): relationship with group B. Journal of Bacteriology, 173: 4814-4819
Liu, W. Liu, Yang, Y. Yang, Chung, N. Chung and Kwang, J. Kwang. (2001). Induction of humoral immune re-sponse and protective immunity in chickens against Salmonella enteritidis after a single dose of killed bacte-rium loaded microspheres. Avian Dis., 45: 797-806
Luk, J.M.C. Luk, Kongmuang, U. Kongmuang, Reeves, P.R. Reeves and Lindberg, A.A. Lindberg. (1993). Selective amplification of abequose and paratose synthetase (rfb) by polymerase chain reaction for identification of Sal-monella major serogroups (A, B, C2, and D). Journal of Clinical Microbiology, 31: 2118–2123
Mastroeni, P. Mastroeni and Menager, N. Menager. (2003). Development of acquired immunity to Salmonella. Journal of Medical Microbiology, 52 : 453-n. mising
Menghistu, H.T. Menghistu, Rathore, R. Rathore, Dhama, K. Dhama and Agarwal, R.K. Agarwal (2011). Isolation, Identification and Polymerase Chain Reaction (PCR) Detection of Salmonella Species from Field Materials of Poultry Origin. International Journal of Microbio-logical Research, 2(2) : 135-142
Menghistu, H.T. Menghistu, Rathore, R. Rathore, Dhama, K. Dhama and Agarwal, R.K. Agarwal (2011). Isolation, identification and polymerase chain reaction (PCR) detection of salmonella species from field materials of poultry origin. International Journal of Microbiological Research, 2(2): 135-142
Mir, I.A. Mir, Wani, S.A. Wani, Hussain, I. Hussain, Qureshi, S.D. Qureshi, Bhat, M.A. Bhat and Nishika-wa, Y. Nishikawa. (2010). Molecular epidemiology and in vitro antimicrobial susceptibility of Salmonella iso-lated from poultry in Kashmir. Rev. Sci. Tech. Off. Int. Epiz., 29(3): 677-686
Murugkar, H.V. Murugkar, Rahman, H. Rahman, Kumar, A. Kumar and Bhattacharya, D. Bhattacharya. (2005). Isolation, phage typing and antibiogram of Salmonella from man and animals in northeastern India. Indian Journal of Medical Research, 122: 237-242
Myint, M.S. Myint. (2004). Epidemiology of Salmonella contamination of poultry products; Knowledge gaps in the farm to store products, M.Sc. thesis, Maryland Univ., Faculty of the Graduate School. 459
Park, M.K. Park, Choi, K.S. Choi, Kim, M.C. Kim and Chae, J.S. Chae (2001). Differential diagnosis of Salmonella Gallinarum and S. Pullorum using PCR-RFLP. Journal of . Veterinary Sciences, 2: 213-219
Plant, J. Plant, Glynn, A.A. Glynn and Wilson, B.M. Wilson B.M. (1978). Protective effects of a supernatant factor from Salmonella Typhimurium on Salmonella Typhi-murium infection of inbred mice. Infect. Immun., 22: 125-131
Pomeroy, B.S. Pomeroy and Nagaraja, K.V. Nagaraja (1991). Fowl typhoid. Diseases of Poultry, 9th edition. Iowa State University press, Ames, IA, pp. 87-99
Prakash, B., Suryanarayana, T., Muniyappa, L. and Krish-nappa, G. (2005) . Evaluation of salmonella gallinarum outer membrane protein based enzyme linked immuno-sorbent assay for detecting antibodies in vaccinated and infected chicken. International Journal of Poultry Sci-ence, 4 (4): 222-227
Sahar, A., Zoo El-Fakar and Nagwa, Rabie, S. (2009). Im-munogenic properties of outer membrane proteins of salmonella in chicken. Global Veterinaria, 3 (2): 75-79
Shah, D.H. Shah, Park, J.H. Park, Cho, M.R. Cho, Kim M.C. Kim and Chae J.S. Chae. (2005). Allele specific PCR method based on rfbS sequence for distinguishing Salmonella Gallinarum from Salmonella Pullorum: serotype-specific rfbS sequence polymorphism. Journal of Microbiological Methods, 60: 169-177
Shivaprasad, H.L. Shivaprasad. (2000). Fowl typhoid and pullorum disease. Review in Scientific Technologies (Off. Inter. Des Epi.), 19: 405-424
Sonne-Hansen, J. Sonne-Hansen and Jenabian, S.M. Jenabi-an. (2005). Molecular serotyping of Salmonella: Identi-fication of the phase 1H antigen based on partial se-quencing of the fliC gene. Acta Pathologica, Microbio-logica et Immunologica Scandinavica, 113: 340-348
Timms, L.M. Timms, Marshall, R.N. Marshall and Breslin, M.F. Breslin. (1994). Laboratory and field trial assess-ment of protection given by a Salmonella enteritidis PT4 inactivated, adjuvant vaccine, Br. Vet. J., 150: 93–102
Verma, N. Verma and Reeves, P.R. Reeves. (1989). Iden-tification and sequence of rfbS and rfbE, which determine antigenic specificity of group A and group D Salmonellae. Journal of Bacteriology, 171: 5694-5701
Al-Zenki, S., Al-Nasser, A., Al-Safar, A., Alomirah, H., Al-Haddad, A., Hendriksen, R.S., Aarestrup, F.M., Aarestrup. (2007). Prevalence and antibiotic resistance of Salmonella isolated from a poultry farm and pro-cessing plant environment in the State of Kuwait. Food-borne Pathog Dis., 4(3): 367-373
Bauer, A.W., Bauer, Kirby, W.M., Kirby, Sherris, J.C., Sher-ris and Turck , M., Turck. (1966). Antibiotic suscepti-bility testing by a standardized single disk method. American Journal of Clinical Pathology, 45(4): 493-496
Bouzoubaa, K. Bouzoubaa, Nagaraja, K.V. Nagaraja, Kab-baj, F.Z. Kabbaj, Newman, J.A. Newman and Pomeroy, B.S. Pomeroy (1989). Feasibility of using proteins from Salmonella Gallinarum vs. 9R live vaccine for the pre-vention of fowl typhoid in chickens. Avian. Dis., 33: 385-391
Bouzoubaa, K. Bouzoubaa, Nagaraja, K.V. Nagaraja, New-man, J.A. Newman and Pomeroy, B.S. Pomeroy (1987). Use of membrane proteins from Salmonella Gallinarum for prevention of fowl typhoid infection in chickens. Avian. Dis., 31: 699-704
Carraminana, J.J. Carraminana, Rota, C. Rota, Agustín, I. Agustín and Herrera, A. Herrera (2004). High preva-lence of multiple resistance to antibiotics in Salmonella serovars isolated from a poultry slaughterhouse in Spain. Veterinary Microbiology, 104: 133-139
Choi-Kim, K. Choi-Kim, Maheswaran, S.K. Maheswaran, Felice, L.J. Felice and Molitor, T.W. Molitor (1991). Relationship between the iron regulated outer mem-brane proteins and the outer membrane proteins of in vivo grown Pasteurella multocida. Veterinary Microbi-ology, 28(1): 75-92
Christensen, J.P. Christensen, Olsen, J.E. Olsen and Bis-gaard, M. Bisgaard. (1992a). Characterization of Salmo-nella enterica serotype gallinarum biotypes gallinarum and pullorum by plasmid profiling and biochemical analysis. Avian Pathology, 21: 461 -470
Christensen, J.P. Christensen, Olsen, J.E. Olsen and Bis-gaard, M. Bisgaard (1992b). Ribotypes of Salmonella enterica serovar Gallinarum biovars Gallinarum and Pullorum. Avian Pathology, 22: 725–738
Davies, R.H. Davies and Breslin, M. Breslin (2003). Obser-vations on Salmonella contamination of commercial laying farms before and after cleaning and disinfection. Veterinary Records, 152: 283-287
Desai, A.R., Shah, D.H., Shringi, S., Lee, M.J., Li, Y. H., Cho, M.R., Park, J.H., Eo, S.K, Lee, J.H,, Chae, J. S. (2005). An allele-specific PCR assay for the rapid and serotype-specific detection of Salmonella Pullorum. Avian Dis., 49(4): 558-61
Gast, R.K. Gast (1997). Salmonella infections in Diseases of Poultry. Iowa State University Press, Ames, pp. 81-121.
Holt, J.G Holt, Krieg, N.R. Krieg, Sneath, P.H.A. Sneath and Williams, S.T. Williams. (1994). Bergey’s manual of determinative bacteriology, 9th Ed. Lippincott, Wil-liams & Wilkins, Baltimore, Maryland.
Laemmli, U.K. Laemmli. (1970). Cleavage of structural proteins during the assembly of the head of bacterio-phage T4. Nature, 227(5259): 680–685
Lee, Y.J. Lee, Kim, K.S. Kim, Kwon, Y.K. Kwon and Tak, R.B. Tak. (2003). Biochemical characteristics and anti-microbials susceptibility of Salmonella Gallinarum isolated in Korea. Journal of Veterinary Sciences, 4(2): 161-166
Lee, Y.S. Lee, Chol, K.S. Chol, Kim, M.C. Kim, Han, J.C. Han, Park, J.H. Park, Shah, D.H. Shah and Chae, J.S. Chae. (2005). Purification of outer membrane protein (OMP C) from Salmonella Gallinarum for characteriza-tion of its immunogenicity. Indian Journal of Animal Sciences, 75(2): 164-167
Liu, D., Liu, Verma N.K., Verma, Romana, L.K. Ro,mana and Reeves, P.R. Reeves. (1991). Completing the se-quence of the rfb region of Salmonella serovars typhi (group D) and paratyphi (group A): relationship with group B. Journal of Bacteriology, 173: 4814-4819
Liu, W. Liu, Yang, Y. Yang, Chung, N. Chung and Kwang, J. Kwang. (2001). Induction of humoral immune re-sponse and protective immunity in chickens against Salmonella enteritidis after a single dose of killed bacte-rium loaded microspheres. Avian Dis., 45: 797-806
Luk, J.M.C. Luk, Kongmuang, U. Kongmuang, Reeves, P.R. Reeves and Lindberg, A.A. Lindberg. (1993). Selective amplification of abequose and paratose synthetase (rfb) by polymerase chain reaction for identification of Sal-monella major serogroups (A, B, C2, and D). Journal of Clinical Microbiology, 31: 2118–2123
Mastroeni, P. Mastroeni and Menager, N. Menager. (2003). Development of acquired immunity to Salmonella. Journal of Medical Microbiology, 52 : 453-n. mising
Menghistu, H.T. Menghistu, Rathore, R. Rathore, Dhama, K. Dhama and Agarwal, R.K. Agarwal (2011). Isolation, Identification and Polymerase Chain Reaction (PCR) Detection of Salmonella Species from Field Materials of Poultry Origin. International Journal of Microbio-logical Research, 2(2) : 135-142
Menghistu, H.T. Menghistu, Rathore, R. Rathore, Dhama, K. Dhama and Agarwal, R.K. Agarwal (2011). Isolation, identification and polymerase chain reaction (PCR) detection of salmonella species from field materials of poultry origin. International Journal of Microbiological Research, 2(2): 135-142
Mir, I.A. Mir, Wani, S.A. Wani, Hussain, I. Hussain, Qureshi, S.D. Qureshi, Bhat, M.A. Bhat and Nishika-wa, Y. Nishikawa. (2010). Molecular epidemiology and in vitro antimicrobial susceptibility of Salmonella iso-lated from poultry in Kashmir. Rev. Sci. Tech. Off. Int. Epiz., 29(3): 677-686
Murugkar, H.V. Murugkar, Rahman, H. Rahman, Kumar, A. Kumar and Bhattacharya, D. Bhattacharya. (2005). Isolation, phage typing and antibiogram of Salmonella from man and animals in northeastern India. Indian Journal of Medical Research, 122: 237-242
Myint, M.S. Myint. (2004). Epidemiology of Salmonella contamination of poultry products; Knowledge gaps in the farm to store products, M.Sc. thesis, Maryland Univ., Faculty of the Graduate School. 459
Park, M.K. Park, Choi, K.S. Choi, Kim, M.C. Kim and Chae, J.S. Chae (2001). Differential diagnosis of Salmonella Gallinarum and S. Pullorum using PCR-RFLP. Journal of . Veterinary Sciences, 2: 213-219
Plant, J. Plant, Glynn, A.A. Glynn and Wilson, B.M. Wilson B.M. (1978). Protective effects of a supernatant factor from Salmonella Typhimurium on Salmonella Typhi-murium infection of inbred mice. Infect. Immun., 22: 125-131
Pomeroy, B.S. Pomeroy and Nagaraja, K.V. Nagaraja (1991). Fowl typhoid. Diseases of Poultry, 9th edition. Iowa State University press, Ames, IA, pp. 87-99
Prakash, B., Suryanarayana, T., Muniyappa, L. and Krish-nappa, G. (2005) . Evaluation of salmonella gallinarum outer membrane protein based enzyme linked immuno-sorbent assay for detecting antibodies in vaccinated and infected chicken. International Journal of Poultry Sci-ence, 4 (4): 222-227
Sahar, A., Zoo El-Fakar and Nagwa, Rabie, S. (2009). Im-munogenic properties of outer membrane proteins of salmonella in chicken. Global Veterinaria, 3 (2): 75-79
Shah, D.H. Shah, Park, J.H. Park, Cho, M.R. Cho, Kim M.C. Kim and Chae J.S. Chae. (2005). Allele specific PCR method based on rfbS sequence for distinguishing Salmonella Gallinarum from Salmonella Pullorum: serotype-specific rfbS sequence polymorphism. Journal of Microbiological Methods, 60: 169-177
Shivaprasad, H.L. Shivaprasad. (2000). Fowl typhoid and pullorum disease. Review in Scientific Technologies (Off. Inter. Des Epi.), 19: 405-424
Sonne-Hansen, J. Sonne-Hansen and Jenabian, S.M. Jenabi-an. (2005). Molecular serotyping of Salmonella: Identi-fication of the phase 1H antigen based on partial se-quencing of the fliC gene. Acta Pathologica, Microbio-logica et Immunologica Scandinavica, 113: 340-348
Timms, L.M. Timms, Marshall, R.N. Marshall and Breslin, M.F. Breslin. (1994). Laboratory and field trial assess-ment of protection given by a Salmonella enteritidis PT4 inactivated, adjuvant vaccine, Br. Vet. J., 150: 93–102
Verma, N. Verma and Reeves, P.R. Reeves. (1989). Iden-tification and sequence of rfbS and rfbE, which determine antigenic specificity of group A and group D Salmonellae. Journal of Bacteriology, 171: 5694-5701
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Isolation and characterization of outer membrane proteins (OMPs) from Salmonella Gallinarum in chicken and antibiogram of the isolates. (2016). Journal of Applied and Natural Science, 8(4), 2292-2297. https://doi.org/10.31018/jans.v8i4.1127
