Inhibition of Staphylococcus aureus by Osmorhiza longistylis Root Extracts

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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

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Osmorhiza longistylis

• Smooth Sweet Cicely
• Anise-scented
• Habitat moist woods
• Plant Height 30-90 cm
• Flower Color white
• Blooms May-June
• Native

Source USDA
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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)

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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

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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

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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 37C
• Zones of inhibition measured and compared w/DMSO
  controls

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Results

• Root Extract

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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 37C
• Incubation mixtures diluted 10,000-fold and
  1,000,000-fold and spread plates prepared with
  100 microliters of S. aureus

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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

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Results
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Future Plans

• Dose-response curve of root extracts
• Fractionate root extract using HPLC and assay
  fractions for antibacterial activity
• Isolate and Identify active ingredient

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Acknowledgment

• Paul Evans collected the plants used in this study