Detection of Salmonella spp. antigens in Tolima poultry products by Western Blot

Abstract

Salmonella spp., is a Gram-negative bacterium transmitted to human by consumption of contaminated water and food consumption, mainly poultry products like eggs and chicken meat. The aim of this study was to implement the Western Blot technic to detect the presence of antigenic proteins of Salmonella spp., in chicken carcasses and egg surface and if possible to compare it with the traditional microbiological isolation. A total of 18 chicken carcasses and 18 eggs were collected from the 13 communes and 5 marketplaces of Ibagué city. A chicken carcass and egg surface washes were obtained and an aliquot from each one was processed by using standard international guidelines ISO 6579-1:2017 for bacterial isolation. Another aliquot was centrifuged and the pellet separated by polyacrylamide gel electrophoresis (SDS-PAGE). Separated proteins were transferred in to a nitrocellulose membrane by Western blot. Primary antibody was an in-house rabbit polyclonal anti-Salmonella enteritidis antiserum and the secondary antibody was a goat anti-rabbit IgG conjugated with alkaline phosphatase. The reaction of the two antibodies was detected with the addition of the BCIP-NBT enzyme substrate and the image recorded with a digital camera. Antigenic bands of Salmonella spp. of 10, 15, 17 and 40 kDa and 10, 17, 25, 37 and 75 kDa were detected in 15 out of 18 (83,3 %) and 4 out of 18 (22,2 %) samples from chicken carcasses and egg surface respectively. A total of 4 out of 36 samples were positive to Salmonella spp., by microbiological isolation. It is concluded that the SDS-PAGE and Western blot technic can successfully detectSalmonella antigens in chicken carcasses and egg surface and it may constitute a valuable complementary tool for the detection of this microorganism in poultry products.

Keywords:

aislamiento microbiológico

anticuerpos

diagnóstico

electroforesis

proteínas

antibodies

bacteria

diagnosis

electrophoresis

isolation

proteins

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