According to CLSI guidelines, Mueller Hinton broth is suitable for all non-fastidious bacterial that grow aerobically.

Regards

Hii

Muhammad Amir Khan. I did MIC test by microdilution method using resazurin dye as marker of cellular viability. I used Muller Hinton broth  and got excellent results.So as per my experience and experiment done by me i willl also suggest you to use MHB to take out your experiment.

thanks.

Please refer following abstract. According to our results, Planktonic  S. aureus achieved their maximum growth with BHI. We used BHI for determination of susceptibility of S. aureus to various nano particles and obtained excellent results.

Dear Gayan Kanchana Wijesinghe. Thank you for sharing your valuable suggestion and the abstract. Your study shows promising results with BHI so I will definitely try it, however, I am anticipating slightly different results in my case, as I am dealing with clinical isolates rather than planktonic aggregates.

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Preferred media is Mueller Hinton Broth (MHB). However, Nutrient broth (NB) also fine.

Dear Dr. Muhammad 

Accoding with CLSI M45-A2 "Methods for antimicrobial dilution and disk susceptibility testing of frequentlly isolated or fastidious bacteria; approved guideline" you should use Müeller Hinton Broth supplemented with cations. EUCAST, also recommend to use MH broth.

http://www.eucast.org/ast_of_bacteria/media_preparation/

Hi!! i have measured antimicrobial activity with MTT compound, and i used Müeller Hinton Broth , and also other Nutrient broth, i had not any problem.

Hi !

Although it is in French I hope to help you.

According to CLSI guidelines, Mueller Hinton broth is suitable for all non-fastidious bacterial that grow aerobically.

Regards

I suppose the medium would depend on which compounds you intend to test. Mueller-Hinton broth is the correct base for all staphylococci susceptibility testing and is absolutely required in clinical lab testing (as provided by several excellent references to CLSI/EUCAST). BHI may give higher cell densities but MIC values have not been validated / correlated with clinical efficacy data so the values can not be interpreted. Likewise, excess growth in BHI may give misleading results due to inoculum effects at high densities and bypass the effects of antibiotics that interfere with metabolite synthesis (trimethoprim, sulfamethoxazole..). BHI is a great medium for many purposes (I use it in lab daily), but not for susceptibility assays.

In terms of supplementation:

• MHB+2% NaCl is often recommended when testing beta lactams
• Supplementation to 25 mg/L Ca2+ / 12.5 mg/L Mg2+ is recommended in general but supplementation to 50 mg/L Ca2+ is required when testing calcium dependent antibiotics (such as daptomycin).

Two last considerations:

• The inoculum is important. Try to target an inoculum of 10^6 cfu/mL equivalent to 1:100 of a 0.5 McFarland standard (approx. 1:100 of 0.15 OD600 units)
• Consider adding the resazurin after 24 hours of growth and allow it to incubate for 30-60 minutes. Bacteria can reduce resazurin to resorufin as they are inactivated by antibiotic giving false-positives. Furthermore, there can be background decomposition of resazurin in growth media resulting in a slow fade-to-pink over time even in the absence of bacteria.

Hope that helped - good luck!

MH broth and BHI. MH is recommended. 

Muller-Hinton broth is recommended as per CLSI guidelines. You can also use Brain Heart infusion also. BHI is used for fastidious organisms and MHB is used for non-fastidious organisms. You can check CLSI  (in USA) or EUCAST (in UK) guidelines for further clarification.

I recommend to see CLSI ( Clinical & Laboratory Standards Institute: CLSI Guidelines) to check out which is the best culture media for this. Also, to follow up their procedures.

Consult CLSI guidelines, MH Broth is one preferred medium.

Hello, Muhammad,

CLSI and EUCAST guidelines recommend MH broth - Mueller Hinton Broth.

A lot of literature on AST are replicated through MH broth. This medium d is generally supportive of many bacterial species.

Hope this helps.

For reference:

http://www.eucast.org/clinical_breakpoints/

https://clsi.org/standards/products/microbiology/documents/m100/

Best,

Arizaldo