I have indicated some pathogenic genes of P aeruginosa identified by genome sequencing in a paper, where the reviewer wanted to do a functional study for the genes which I claimed, but I have mentioned multiple genes responsible for virulence and pathogenesis of P aeruginosa which is present in other strains to, mainly exotoxin U and A. can anyone suggest a protocol with out animal study (because animal study not possible in my lab scenario) to validate the strains pathogenicity and virulence factors of P aeruginosa.
Can you perform the cell culture, which specific to the bacteria and function of the cell that you want to study. Then you infect the cell line with your interesting bacteria among various condition of the bacteria or the environment by point of time or amount of bacteria. You should to concern about various confounding factors that can occur in your experiment. May you need to duplicate of the test or repeat the test within day or between day. After that, you observe changing of gene expression (up or down regulate) or secretion of particular protein (increase or decrease), which relate to the pathogenesis of the bacteria, which express or secrete from the cell line.
Hello Nandagopal. I agree with Ditthawat. You can go for cell culture pathway, instead of the animal model. When I read about Pseudomonas aeruginosa and exotoxin U (as attached), they mentioned that the model used was mouse embryonic fibroblasts. So you can potentially do your study using MEF cell line (CF-1) (ATCC® SCRC-1040™), from ATCC.
Article Apoptosis induced by Pseudomonas aeruginosa: A lonely killer?
Pathogenesis defines the capacity to induce or maintain disease. Disease is confined to organisms. Therefore, in the end after having defined virulence factors in vitro, it will not be possible to perform animal experimentation. Sorry for that, but it is unavoidable.
Definitely you can! You can check the virulent factors like RBC haemolysis, Presence of extracellular enzymes etc....
Study the effect of haemolysin on sheep blood agar, effect on tissue culture
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