I have got following results can you please tell me my result are fine or not if not what I should do to improve my result. Moreover, I haven't seen inhibition zone in that experiment. 10-6 I have got 160 colony calculated as 0.062 is it correct calculation??
Hi Alia Alia , to check MIC its better not to go with absorbance. You can perform fluorometric methods which are way more trustworthy. Why don't you try Resazurin assay or MTT assay? That could be a better way to determine bacterial viability against the antibacterial agent.
The image that you added: could you attach another image with your control groups? (positive/negative/blank/media control)
That can help me to give you a better justification.
Hi Alia Alia , the Amp OD600 result doesn't make any sense. If 50 ng/mL is better than 6.25 ng/mL and have better antimicrobial property, the absorbance should be way lower. In your case, its the opposite. You controls have problems too. Negative control should be where there is microbial growth, so you should get high OD value. Positive control means it should have an agent which kills bacteria, so it should have the least OD value.
I think you should repeat the whole experiment. Check for pipetting errors, contaminations, and labeling errors. As I suggested before, do fluorometric methods to check bacteria viability... don't go for OD, because sometimes even agents show absorbance and they interfere in the density check.
To check cfu/mL, carry out a 4 or 5 fold dilution and then pour 10 ul in MHA plates. You may also have to increase the concentration of the agents you are using.
When you get a data that is similar to your media control (only media, no bacteria, no drug), and positive control (media, drug that inhibited bacteria totally), then you can say that it is the MIC of the tested agent.
Good luck.
You may go through the following articles: It will help you.
Article Prevalence and mechanisms of antibiotic resistance in Escher...
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https://medwinpublishers.com/AABSc/antimicrobial-assessment-of-a-novel-humectant-hand-sanitizer-against-microbes-transferred-to-human-palms-from-mobile-phones-with-without-flip-covers.pdf
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