Methods for in vitro evaluating

antimicrobial activity

Broth dilution methods (liquid media)

Agar diffusion methods (solid media)

 About MIC and MBC

The  range  of  an+bio+c  concentra+ons  used  for  determining  MICs  is  universally  accepted  to  be  
in   doubling   dilu*on   steps   up   and   down   from   1   mg/L   as   required.   The   MIC   is   defined   as   the  
lowest   concentra+on   of   a   drug   that   will   inhibit   the   visible   growth   of   an   organism   a?er  
overnight  incuba+on  (this  period  is  extended  for  organisms  such  as  anaerobes,  which  require  
prolonged  incuba+on  for  growth).  
 Broth dilution method
(microdilution = using microtiter plates)
About MIC and MBC (liquid
About MIC and NIC
 About MIC and NIC

MIC  is  a  difficult  subject:  

 
-­‐  It’s  a  RANGE  of  concentra+ons  depending  on  the  
dilu+on  series  used  

-­‐  Comparison  of  values  obtained  from  the  

mul+tude  of  different  techniques  is  problema+c  

-­‐  MIC  and  NIC  are  dependent  on  the  experimental  

condi+ons:  

-­‐  Incuba+on  temperature  

-­‐  Microorganism  
-­‐  Inoculum  size  
-­‐  Incuba+on  +me  
-­‐  Growth  medium  
-­‐  etc.  
Lambert  and  Pearson,  2000.  J  Appl  Microbiol  88:784  
Use  only  pure  cultures  !!  
 Agar diffusion methods !

Agar-disc diffusion method:

diffusion of antibiotic! Antimicrobial gradient method!
from a paper disc! i.e., E-test® commercial strips !