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METHODOLOGY

Bacterial strains

One Gram-positive (Staphylococcus aureus) and two Gram-negative (Pseudomonas


aeruginosa and Klebsiella pneumonia) bacteria were selected for antibacterial activity assay.

Antimicrobial activity assay by disc diffusion method

The peppermint extract was dissolved in ethanol to a final concentration of 1 mg/ml. The
bacterial strains were grown on nutrient agar (NA) by spread plate method. The bacteria were
pipetted onto surface of agar plate (0.1 ml or less) and were spread evenly over surface of
agar using sterile glass spreader. Blank discs were impregnated with peppermint extract using
ethanol. The discs were allowed to dry at room temperature before placed on the agar plate. A
disc impregnated with distilled water was used as a negative control. Bacitracin disc,
cefepime disc and ampicillin disc was used as a positive reference standard to determine the
sensitivity of the bacterial strains respectively. The plates were incubated at 37°C for 24 h.
Antimicrobial activity was evaluated by measuring the diameter zone of inhibition around the
disc. The resultant clear zones around the discs were measured in mm. The antibacterial
activity of peppermint extracts were indicated by clear zones of growth inhibition.

REFERENCES

Bupesh, G., Amutha, C., Nandagopal, S., Ganeshkumar, A., Sureshkumar, P., & Murali, K.
(2007). Antibacterial activity of Mentha piperita L. (peppermint) from leaf extracts - a
medicinal plant. Acta Agriculturae Slovenica, 89(1), 73-79. doi:10.2478/v10014-007-
0009-7

Basheer, A. A., & Abdullah, A. A. (2013). Antimicrobial activity of the aqueous extract of
mint plant. Science Journal of Clinical Medicine, 2(3), 110-113.
doi:10.11648/j.sjcm.20130203.19

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