Oil of Oregano Effects on Oil of Oregano Effects on Microbial FloraMicrobial Flora

Cameron Herbst

Pittsburgh Central Catholic High School

The Claim

Hailed as a highly potent purifier, antiviral, antibacterial, antifungal, antiparasitic.

Benefits respiratory health, joint/muscle flexibility, destroy organisms that contribute to skin infections/digestive problems, strengthen immune system

The Claim

Countless studies have evaluated the affect of ingested material on human cells and physiology.

A key component of the body is the natural flora - especially those within the alimentary canal. These belong to the families: Clostridium, Fusobacterium, Escherichia, Bacteroides, Eubacterium, and Lactobacillus, among others.

Oil of OreganoOil of OreganoDerived from wild oregano:

Origanum vulgareTwo key compounds: carvacrol,

thymol (studies suggest may combat microorganisms which cause illness)

Ingredients: 60% carvacrol (3.2 mg), olive oil

Production and Uses of Oil of OreganoProduction and Uses of Oil of Oregano

Marketed as “supports general well-being”

Obtained from crushing the leaves of the wild oregano plant

Dosage is 2 dropsCan be ingested or applied

topically

Risks of Oil of Oregano UseRisks of Oil of Oregano Use

Reduces the body’s ability to absorb iron

Not advisable for pregnant women, stimulates blood flow in the uterus, weakening lining of fetus in the womb

Can cause skin irritation, rashes, or vomiting if you have allergies to thyme, basil, mint, or sage

E.E. colicoliOne of the most common forms of

bacteria; Free living, symbiotic or pathogenic

Has been utilized as the most studied prokaryote

There are many of different strains of E. coli, most of which are non-pathogenic. However, there are strains which can produce fatal diseases

Staphylococcus epidermidis

Part of our normal/skin flora, gram-positiveAnaerobic, but grows best in aerobic

conditionsOpportunistic pathogen, requires major

breach in hosts defensesNon-pathogenic strain utilized

PurposePurposeTo determine whether oil of

oregano in various concentrations will affect the survivorship of intestinal flora (E. coli) and Staphylococcus epidermidis.

HypothesisHypothesis

Null – Survivorship of E. coli and Staphylococcus epidermidis in varying concentrations of oil of oregano would not vary significantly from the control.

Alternative – Survivorship of E. coli and Staphylococcus epidermidis in varying concentrations of oil of oregano would vary significantly from the control.

MaterialsMaterials Ethanol (sterilization of instruments) Latex gloves E. coli DH5 alpha Micropipette Tube rack 8 Tubes SDF (per 1 liter) (100mM KH2PO4, 100mM K2HPO4, 10mM MgSO4, 1mM NaCl) Turn table LB agar plates LB media (0.5% yeast extract, 1% tryptone, 1% sodium chloride) Bunsen burner Spreader bars Matches Sterile pipette tips Incubator Vortex Klett spectrophotometer Staphylococcus epidermidis

ProcedureProcedure1) E. coli and Staphylococcus epidermidis was grown

overnight in sterile LB media.2) A sample of the overnight culture was added to

fresh media in a sterile sidearm flask.3) The cultures were placed in an incubator (37°C)

until a density of 50 Klett spectrophotometer units was reached. This represents a cell density of approximately 108 cells/mL.

4) The culture was diluted in sterile dilution fluid to a concentration of approximately 103 cells/mL.

5) The tubes were prepared as follows:

ProcedureProcedureTube # Oil of

Oregano (mL)

SDF (mL) E. coli/Staph

(mL)

Final Volume

(mL)

Final Concentration

(%)

1 (E. coli) 0 9.9 0.1 10 0

2 (E. coli) 0.01 9.89 0.1 10 0.1

3 (E. coli) 0.1 9.8 0.1 10 1

4 (E. coli) 0.5 9.4 0.1 10 5

1 (Staph) 0 9.9 0.1 10 0

2 (Staph) 0.01 9.89 0.1 10 0.1

3 (Staph) 0.1 9.8 0.1 10 1

4 (Staph) 0.5 9.4 0.1 10 5

ProcedureProcedure6) The E. coli and Staphylococcus epidermidis

was allowed to remain exposed to the oil of oregano for 15 minutes.

7) 100 µL aliquots were removed from the tubes and spread on LB plates.(6 replicates)

8) The plates were incubated at 37 degrees Celsius for 24 hours.

9) The resulting colonies were quantified. Each colony is assumed to have arisen from one cell.

E. coli Oil of Oregano Survivorship Graph

0

10

20

30

40

50

60

70

80

90

0 0.1 1 5

Concentration (%)

Num

ber o

f Col

onie

s

E. coli Colonies

p= 9.63E-8

Staph Oil of Oregano Survivorship Graph

0

20

40

60

80

100

120

140

160

0 0.1 1 5

Concentration (%)

Num

ber

of C

olon

ies

Staph Colonies

p= 6.19E-4

E.E. colicoli Dunnett’s Test Dunnett’s Test

Variable Comparison t-value Interpretation

0.1% vs control 67.75 Significant

1% vs control 53.41 Significant

5% vs control 23.75 Significant

α = 0.01t-crit = 4.21

Staph Dunnett’s Test α = 0.01t-crit = 4.21

Variable Comparison t-value Interpretation

0.1% vs control 133.70 Significant

1% vs control 122.70 Significant

5% vs control 95.20 Significant

InterpretationInterpretationThere appeared to be a correlation

between the percent concentration of oil of oregano and cell survivorship. Higher concentrations of oil of oregano resulted in fewer surviving colonies in both Staphylococcus epidermidis and E. coli.

ConclusionConclusion5% oil of oregano appeared to greatly

reduce E. coli survivorship

The null hypothesis can be rejected for all concentrations of oil of oregano for E. coli and Staphylococcus epidermidis.

LimitationsLimitationsThis experiment was limited in that

pure oil of oregano was not able to be used.

Plating was not exactly synchronized, which could have resulted in extra time for bacterial replication.

ExtensionsExtensionsVarying exposure time of E. coli and

Staphylococcus epidermidis in the tubes before plating will be tested.

To allow for longer exposure, oil of oregano will be infused directly into the LB agar.

A different symbiotic organism other than E. coli and Staphylococcus epidermidis will be tested.

Cited SourcesCited Sourceshttp://e-colibasics.com/www.oiloforegano.com/http://randi.org/site/index.php/swift-blo

g/1099-oil-of-oregano-miracle-cure.html

www.fda.govhttp://web.uconn.edu/mcbstaff/graf/Stu

dent%20presentations/S%20epidermidis/sepidermidis.html