The aims of this research are confirming the feasibility of hybrid immunoglobulin-binding reagents, its used in ELISAs for IgG/IgY detection and detecting specific antibodies against an infectious microorganism (Salmonella spp) in various animal species using a universal diagnostic ELISA. Hybrid immunoglobulin-binding bacterial proteins (IBP) including recombinant protein LA, recombinant protein LG, and recombinant protein AG have been produced for improvement of their binding affinity to a much larger number of immunoglobulins. This hybrid bacterial protein thus represents a powerful tool for binding, detection, and purification of immunoglobulins and their fragments. However, SpLA-LG-peroxidase and SpLAG-anti-IgY-peroxidase were produced by the periodate method. They have shown to be effective reagents. Their binding affinity to immunoglobulins surpasses previous hybrid IgG-binding proteins reported, including the most known SpAG, SpLA and SpLG. The IgY fraction was isolated from the egg yolks of a variety of birds including species of chicken, bantam hen, guinea hen, quail, goose, duck, wild and domestic pigeon, parakeet, cattle egret, pheasant, and ostrich. The IgY fraction was isolated by the chloroform-polyethylene glycol (PEG) method. An ELISA for anti-Salmonella spp antibodies was employed with some modifications to determine the presence of antibodies in humans, laying hens, geese, quails, and pigeons. Salmonella are motile, flagellated rod-shaped zoonotic pathogens which may survive with or without oxygen. They belong to the family Enterobacteriaceae and is implicated with typhoid fever and food-borne illnesses. This pathogen is associated with several diseases, which may become fatal and negatively impact the health of individuals and various economies globally. The poultry industry is most impacted and vulnerable to the onslaught of this pernicious microbe.
The aim of this study was to develop a natural cost effective method of bacterial attenuation using indigenous plants to produce a safe live attenuated vaccine for poultry salmonellosis. Several indigenous plants were used but garlic and onion gave the expected result of Salmonella attenuation. Chicken were immunized with a attenuated Salmonella cocktail (mixture of five Salmonella strains: S. Montevideo, S. Yeerongpilly, S. Augustenborg, S. Kentucky, and S. Typhimurium). ELISA for Salmonella antibodies showed high titer suggesting that the indigenous plant did not affect the immunogenic capability of the vaccine. Garlic and onion can be considered as natural bacteriostatic agents that inhibited the growth of Salmonella in vitro contributing to a safe and effective vaccine against poultry salmonellosis.
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