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Inhibition of Staphylococcus aureus by Osmorhiza longistylis Root Extracts. Jason N. Herold, Jennifer Antisdel, Bernadette Corbett, Del Stallwood Metropolitan Community College Omaha Nebraska . Smooth Sweet Cicely Anise-scented Habitat: moist woods Plant Height: 30-90 cm
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Inhibition of Staphylococcus aureusby Osmorhiza longistylis Root Extracts Jason N. Herold, Jennifer Antisdel, Bernadette Corbett, Del Stallwood Metropolitan Community College Omaha Nebraska
Smooth Sweet Cicely Anise-scented Habitat: moist woods Plant Height: 30-90 cm Flower Color: white Blooms: May-June Native Osmorhiza longistylis 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
Results Root Extract
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
Future Plans • Dose-response curve of root extracts • Fractionate root extract using HPLC and assay fractions for antibacterial activity • Isolate and Identify active ingredient
Acknowledgment • Paul Evans collected the plants used in this study