1. The Effects of Ginseng on Microflora Survivorship
Joseph Fantini
11th grade, Pittsburgh Central Catholic High School,
2nd year in PJAS

2. Problem
Ginseng is widely known for its use in folk medicine; medicine that is not always scientifically explicable
Developed as folk medicine without understanding effect on microbial life

3. Potential mutualistic, parasitic, pathogenic, or commensal effects
Microbial Flora
Internal and external flora include eukaryotic fungi, protists, and bacteria
Little is understood about the association between humans and their flora
Potential mutualistic, parasitic, pathogenic, or commensal effects
Provides nutritional and digestive benefits, secrete vitamins, stimulate antibody production, and protect against pathogenic microbes
Ingested substances may have an impact on colony sizes and functions of flora

4. Escherichia Coli (E. Coli)
Large and diverse group of gram (-) bacteria
Free living, symbiont, or pathogen
Live in the intestinal tract of many mammals
Most strains are not pathogenic
Serve as a common prokaryotic cell model

5. Staphylococcus epidermidis Widely studied Gram-positive bacterium
Part of microflora found on skin
Multiple positive effects on the body
Inhibition and killing of nonindigenous species

6. Ginseng Capsules (Giant Eagle) Components
Ginseng being tested
Ginseng Capsules (Giant Eagle)
Components
American Ginseng Extract (Panax quinquefolius)
Chinese Red Ginseng Extract (Panax Ginseng)
Panax Ginseng (Panaz Ginseng)

7. Purpose
To determine the effects of Ginseng on two different forms of microbial life—Staphylococcus epidermidis and Escherichia coli

8. Hypothesis
Null hypothesis: Ginseng will not have a significant effect on staph and E. coli survivorship.
Alternate hypothesis: Ginseng will have a significant effect on staph and E. coli survivorship.

9. Materials Escherichia coli Bunsen Burner Staphylococcus epidermidis
Ginseng Capsules
Micropipettes
Sterile Water
Micro Rack
Sterile Pipette Tips
Micro Tubes
60 YEPD Agar plates (1% yeast extract, 2% peptone 2% glucose, 1.5% agar)
Agar Plates
YEPD Media (1% yeast extract, 2% peptone, 2% glucose)
Vortex
Incubator
Sterile dilution fluid (10 mM KH2PO4, 10mM K2HPO4, 1mM MgSO4, 0.1 mM CaCl2, 100 mM NaCl)
Spreader Bars
Ethanol

10. Procedure
Microbes were grown overnight in sterile YEPD media
A sample of the overnight culture was added to fresh media in a sterile sidearm flask
The culture was incubated at 30C until a density of 50 Klett spectrophotometer units was reached. This represents a cell density of approximately 107 cells/mL
The cell culture was diluted in sterile dilution fluid to a concentration of approximately 105 cells/mL
Test tubes were made with concentrations of 0%, 0.1%, 1%, and 10% Ginseng. (5 replicates for each of the four concentrations – 20 plates per microbe)

11. Test Tube Concentrations0%
0.1% 1%
10%
Sterile water
9.9 mL
9.89 mL
9.8 mL
8.9 mL
Microbe
0.1 mL
Ginseng
0 mL
0.01 mL
1 mL

12. Procedure continued
6. The tubes were incubated at room temperature for 5 minutes 7. Tubes were vortexed and 0.1 mL aliquots were plated onto YEPD agar 8. Plates were incubated at 30 degrees Celsius for one day and colonies were counted. Each colony was assumed to have arisen from a single cell.

13. A-Nova Dunnett's Test A-Nova & Dunnett's Test
Statistical test that allows for the comparison of means of different groups to determine significant variation
Dunnett's Test
A follow up test used to find out which variable groups produced significant variation compared to a control

14. P-Value: 5.91789E-11 P-Value: 1.20143E-14 Cell Colonies Counted
Percent Concentration of E. coli and Staph

15. Dunnett’s Test Results
T Value
Result
(E.COLI) 0% vs. 0.1% Ginseng
4.25
Significant
(E.COLI) 0% vs. 1% Ginseng
10.97
(E.COLI) 0% vs. 10% Ginseng
14.95
(STAPH) 0% vs. 0.1% Ginseng
3.81
(STAPH) 0% vs. 1% Ginseng
18.87
(STAPH) 0% vs. 10% Ginseng
24.02
t-crit = 3.239

16. Interpretation of Results
The null hypothesis can be rejected
The alternate hypothesis can be accepted
Ginseng produced a significant negative effect on E. coli and staph survivorship
Statistical analyses supported a significant effect at all concentrations

17. Limitations
Slight positioning differences in the incubation process
Slightly desynchronized plating
Limited concentration exposures
Only one exposure time
Only survivorship tested, not growth or other parameters
Study does not account for other factors that might affect the microbial flora

18. Addition of more microbial models such as Yeast
Extensions
Addition of more microbial models such as Yeast
Would give insight on effects on more advanced cells like human cells
Different Species of Ginseng can be tested in isolation
Different concentrations of the Ginseng could be tested
Agar infusion of Ginseng
Test for synergistic effects with other chemicals

19. Bibliography
anova-statistical-guide.php
support/microbiology/staphylococcus-epidermis/

20. Single Factor A-Nova (E. Coli)

21. Single Factor A-Nova (Staph)