The Pharmacological Effect Of Antibiotics

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The Pharmacological effect of antibiotics.

This practical involved the determination of susceptibility of antimicrobial agents using the Kirby-Bauer sensitivity test as well as investigating the minimal inhibitory concentration of penicillin. This practical also demonstrated the reversal of penicillin inhibition in the presence or absence of β-lactamase.

Method A nutrient agar plate containing the test organism with attenuated enteric bacteria was inoculated by use of a swab dipped into bacterial suspension provided. Inoculation streaks were done on the entire plate after which the agar plate was left to dry for 5 minutes. The filter paper disc impregnated with antibiotics was picked up by a pair of forceps, dipped in alcohol and passed through a flame for sterilization purposes. The filter paper disc were then pressed onto the agar plate using the forceps and incubated for 48hours at 37 degrees Celsius. The pre-incubated annual radii between the edge of the disk and the edge of the confluent were measured and the results recorded after incubation took place.
Test tubes containing serial dilution of 1: 1000 brain-heart infusion (BHI) broth attenuated with strains of the bacteria was incubated overnight at 37 degrees Celsius. Incubation was done in the absence or presence of increasing concentration of penicillin. 200 microliter was pipetted into the 96 well plate after which it was analyzed using a microplate reader using 570nm optical density.

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