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Inhibition of Staphylococcus aureus
by Osmorhiza longistylis
Root Extracts
Jason N. Herold, Jennifer Antisdel, Bernadette Corbett, Del Stallwood
Metropolitan Community CollegeOmaha Nebraska
Osmorhiza longistylis
Smooth Sweet Cicely Anise-scented Habitat: moist woods Plant Height: 30-90
cm Flower Color: white Blooms: May-June Native
Source: USDA
Native American Uses Omaha
Poultice of pounded root applied to boils.
Winnebago Poultice of pounded root applied to wounds.
Gilmore, M. R. 1919 Uses of Plants used by the Indians of the Missouri River Region. SI-BAE Annual Report #33 (p.107)
Preparation of the Extracts Roots, stems and leaves separated Plant material washed with distilled
water, then blotted dry Plant material ground in Waring Blender
in 95% ethanol Plant material filtered off and discarded Ethanol distilled off, leaving an aqueous
sample
Preparation of Extracts
Aqueous samples extracted with methylene chloride
Dried with anhydrous sodium sulfate
Liquid decanted into round bottomed flask
Methylene chloride removed by distillation
Diffusion Disk Method 24 hour cultures of S. aureus plated
on brain heart agar 100 micrograms extract reconstituted
in 20 microliters of DMSO and pipetted onto filter paper disks
Disks placed on plates and incubated for 24 hours at 37°C
Zones of inhibition measured and compared w/DMSO controls
Broth Dilution Method
24 hour Brain-Heart Broth cultures 100 micrograms of plant extract with
20 microliters DMSO incubated with 180 microliters of S. aureus culture for 2 and 24 hours at 37°C
Incubation mixtures diluted 10,000-fold and 1,000,000-fold and spread plates prepared with 100 microliters of S. aureus
Results
Stem and leaf extracts did not inhibit growth when compared to DMSO control
Root extracts inhibited growth by 12.2% and 25% when incubated for 2 and 24 hours, respectively
Results
0
5
10
15
20
25
% Inhibition
Root Stem Leaf
Mean Inhibition Compared To DMSO Control
2 Hour
24 Hour
Future Plans
Dose-response curve of root extracts
Fractionate root extract using HPLC and assay fractions for antibacterial activity
Isolate and Identify active ingredient