A Simple Procedure for Extraction of Surface Protein of Salmonella Serotypes and Escherichia coli Strains Isolated from Poultry and Pigs

Tran Thi Quynh Lan (Department of Veterinary Biosciences, Faculty of Animal Science and Veterinary Medicine Nong Lam University, Ho Chi Minh City)
Doan Thi Da Linh (Faculty of Animal Science and Veterinary Medicine, Nong Lam University, Ho Chi Minh City)
Le Ho Truc Phuong (Faculty of Animal Science and Veterinary Medicine, Nong Lam University, Ho Chi Minh City)

Abstract


Salmonella and E.coli possess different surface protein structures that can induce protective immune responses. Identification of these proteins capacitates development of diverse applications in prevention and diagnosis that contribute to effectively control disease-causing enterobacteria pathogens such as Salmonella and E.coli. A simple procedure for obtaining protein complexes of Salmonella serotypes and E.coli is performed in this study. A sonication process with heat treatment of whole bacteria induced the release of protein complexes. Concentration of the protein extract was quantified using protein quantification Kits-Rapid, and protein complex profile was obtained by SDS-PAGE (Sodium dodecyl sulfate polyacrylamide gel electrophoresis) and silver staining. The concentrations of protein ranged from 29.45 to 45.35 µg/mL in the  Salmonella protein extracts, and from 25.35 to 36.72 µg/mL in the E.coli protein extracts. Six major groups of proteins from E. coli (YfiO, NipB, OmpF, YfgL, Talc, YaeT) and four major groups of proteins from Salmonella (Flagellin, OmpA, Porin, SEF21) were preliminarily determined by a simple procedure of extraction based on the molecular weight.


Keywords


Salmonella serotypes; E.coli strains; Extraction; Protein complexes extract; SDS-PAGE

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References


[1] Giannella RA. Salmonella. In S. Baron, Medical microbiology, University of Texas Medical Branch at Galveston, Galveston, Texas, 1996, 4th.

[2] Quinn PJ, Markey BK, Leonard FC, Hartigan P, Fanning S and Jayaprakash NS. Veterinary Microbiology and Microbial Disease, III. 2nd edition. Willey Blackwell, USA, 2011: 263-287.

[3] Edens FW. An alternative for antibiotic use in poultry: probiotics, Department of Poultry Science of North Caroline State University, December 10th, 2015.

[4] http://www.scielo.br/scielo.php?script=sci_arttext&pid=s1516-635x2003000200001#end

[5] Tizzard IR. Veterinary immunology. W.B. Saunders Co.,United States, 2004: 272-280, 7th edition.

[6] Singh K and Lahiri T. An improved protein surface sxtraction method using rotating cylinder probe. Interdisciplinary Sciences: Computational Life Sciences, 2017, 9 (1): 65-71.

[7] Padmanabhan V, Govind GA, Kamalanathan AS and Yayprakash NS. Extraction of Antigenic Membrane Proteins from Salmonella Using Detergent and Phase Partition Method. Res. J. Microbiol, 2018, 13 (1): 47-52.

[8] Tran TQ Lan, Marie-Lou G, Nguyen TM. Nhan, Letellier A and Quessy S. Distribution of Virulence Genes among Salmonella Serotypes Isolated from Pigs in Southern Vietnam. J. Food Protect., 2018, 81(9): 1459-1466.

[9] Tran TQ Lan, Nguyen TM Nhan, Nguyen T Huyen. Survey on the prevalence of Salmonella group and serotype isolated from pig and poultry feces. J. Vet. Sci. & Technol. 2013, (In Vietnamese) XX (3): 30-36.

[10] Tran TQ Lan, Lewandowski SL, Quessy S, Letellier A and Boulianne M. Identification of Immunoreactive Antigens of Salmonella Enteritidis, Processding of 8th science conference at Nong Lam University, 2011.

[11] Ochoa-Reparaz Javier, Begona Sesma, Miguel Alvarez, M. Jesus Renedo, Juan Irache, Carlos Gamazo. Humoral immune response in hens naturally infected with Salmonella Enteritidis against outer membrane proteins and other surface structural antigens. EDP sciences Vet. Res. 2004, 35: 291-298.

[12] Stenberg F, Chovanec P, Maslen SL, Robinson CV, Ilag LL, von Heijne G and Daley DO. Protein Complexes of the Escherichia coli Cell Envelope. J. Biol. Chem, 2005, 41: 34409-34419.

[13] Lasserre JP, Beyne E, Pyndiah S, Lapaillerie D, Claverol S and Bonneu M. A complexomic study of Escherichia coli using two-dimensional blue native/ SDS polyacrylamide gel electrophoresis. Electrophoresis, 2006, 27: 3306-3321.



DOI: https://doi.org/10.30564/vsr.v1i2.1357

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