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Agar dilution is one of two methods (along with broth dilution) used by researchers to determine the minimum inhibitory concentration (MIC) of antibiotics. It is the dilution method most frequently used to test the effectiveness of new antibiotics when a few antibiotics are tested against a large panel of different bacteria. [1]: 149 [2]
The minimum inhibitory concentration, which is the lowest concentration of the antibiotic that stops the growth of bacteria, can be estimated from the size of the zone of inhibition. Antibiotic susceptibility testing has been needed since the discovery of the beta-lactam antibiotic penicillin. Initial methods were phenotypic, and involved ...
Ampicillin is used to treat infections by many gram-positive and gram-negative bacteria. It was the first "broad spectrum" penicillin with activity against gram-positive bacteria, including Streptococcus pneumoniae , Streptococcus pyogenes , some isolates of Staphylococcus aureus (but not penicillin-resistant or methicillin-resistant strains ...
Blood agar plates (BAPs) contain mammalian blood (usually sheep or horse), typically at a 5–10% concentration. BAPs are enriched, and differential media is used to isolate fastidious organisms and detect hemolytic activity. β-Hemolytic activity will show lysis and complete digestion of red blood cell contents surrounding a colony.
The pour plate technique is the typical technique used to prepare plate count agars. Here, the inoculum is added to the molten agar before pouring the plate. The molten agar is cooled to about 45 degrees Celsius and is poured using a sterile method into a petri dish containing a specific diluted sample.
LB medium bottle and LB agar plate Plate medium agar LB. Lysogeny broth (LB) is a nutritionally rich medium primarily used for the growth of bacteria. Its creator, Giuseppe Bertani, intended LB to stand for lysogeny broth, [1] but LB has also come to colloquially mean Luria broth, Lennox broth, life broth or Luria–Bertani medium. [2]
The minimum bactericidal concentration (MBC) is the lowest concentration of an antibacterial agent required to kill a particular bacterium. [1] It can be determined from broth dilution minimum inhibitory concentration (MIC) tests by subculturing to agar plates that do not contain the test agent.
Plate 1:10 and 1:100 dilutions of the incubated cultures on selective/ screening plates (e.g. Ampicillin and/or X-gal) onto LB plates to which the antibiotics to be used for selection have been added; Incubate overnight at 37°C; Finally, observe isolated colonies on the plates