I want to do research on the synergistic/antagonistic effects of plant extracts and antibiotics against some bacteria. It will be very helpful if you give me literature/detailed methodology about this topic.
Brain heart Infusion (BHI BROTH) is required only for enriching the isolates from clinical samples which are from sterile sites (like blood).
Since you have already have the isolates from clinical samples, if they are usual aerobes, even peptone water would be enough.
If you have already extracted the compound from the plant. You have to follow the Kirby Baeur Disk diffusion method protocol for testing the effect of plant extracts on the usual pathogens. You need to test against few clinical Gram positives (eg. S. aureus) as well as Gram negatives (Eg. E. coli), do not forget to include ATCC bacterial strains as controls (any 2; E. coli and S. aureus). Please follow any recent literature (published paper- in reputed journal) for the disk diffusion protocol. After this you can follow the MIC DETERMINATION PROTOCOL (AGAR DILUTION METHOD).
Thank you for the response. How is the Checkerboard method carried out to evaluate the synergism between plant extracts and antibiotics?
Since they are pathogens, to maintain the virulence/pathogenicity Brain heart Infusion medium is required. If the strains were cultured repeatedly in a medium without any animal source, they may lose their pathogenic nature. So it is always advisable to maintain the culture in Brain heart Infusion broth.
Peptone water does have nutrients of animal source. We are talking about just culturing the isolates for tests to be carried out. We are not talking about stocking the isolates. For stocking, the media has to be enriched and should be with glycerol, to be stored at -20C or -80C. One set (duplicate) should not be moved out of freezer to avoid repeated freezing and thawing. Incase we have freeze drier that would solve all the problems in stocking.
Make sure you pick up isolated definite colonies to inoculate into your liquid media (simple media like peptone) (nutrient broth or BHI broth if fastidious bacteria) to carry out for your further susceptibility tests.
Cheers :)
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