By: Luke BeckPittsburgh Central Catholic HSPJAS 2012Grade 11

Synergistic Drug Effects on Microbial Flora

Accompanied with drug side effects, combination of over the counter drugs can have dire side effects

Do common daily over the counter drugs cause adverse effects on human cells?

Problem

A nonsteroidal anti-inflammatory drug

Used to treat such symptoms as headache, muscular aches, the common cold and arthritis pain.

Third most consumed drug in the entire world.

Can be taken on a daily basis to prevent symptoms.

Ibuprofen

Active Ingredient: Diphenhydramine Hydrochloride

Used as an Antihistamine Over 25 over the counter

drugs share the same active ingredient as Benadryl

Taken on a daily basis to prevent allergy symptoms

Benadryl

Has been the most studied type of bacteria in biological research.

Gram negative bacteria Symbiont in intestinal tracts

of many mammals, including humans.

Mostly non-pathogenic.

Escherichia Coli

Staph. species consisting harmless skin-dwelling microorganisms

Species of Staph. are commonly used in microbiological experiments

Gram Positive Bacteria

Staphylococcus epidermidis

To determine if differing concentrations of Ibuprofen, the antihistamine Benadryl and/or synergy of the two have an adverse effect on E. Coli and Staph. Survivorship.

Purpose

Hypothesis

Null: The presence of Ibuprofen, Benadryl, and a combination of both will not cause a change in bacterial population outside of chance

Alternative: The presence of Ibuprofen, Benadryl, and a combination of both will cause a change in bacterial population outside of chance

Materials

Ibuprofen (200mg liquid gels)

Benadryl (25mg liquid gels) Escherichia coli bacteria Staphylococcus epidermidis

bacteria Ethanol 20 mL tubes Sterile Micro burner Ethanol Test tubes Test Tube Rack Microtubes

Vortex Incubator Gloves Goggles Klett Spectrophotometer SDF LB agar plates (1% Tryptone,

0.5% Yeast Extract, 1% NaCl) Sidearm flask Spreader bar SDF Test Tubes

1. Bacteria (E. coli and Staph) 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 culture was 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 105 cells/mL.

Procedure

Concentrations

(Ibuprofen, Benadryl)

Control

(0 x, 0.01x)

(0 x, 0.1 x)

(0.1 x, 0 x)

(0.1 x, 0.01 x)

(0.1 x, 0.1 x)

(1 x, 0 x)

(1 x, 0.01 x)

(1 x, 0.1 x)

MicrobeE. Coli or Staph.

0.1 mL 0.1 mL 0.1 mL 0.1 mL 0.1 mL 0.1 mL 0.1 mL 0.1 mL 0.1 mL

SDF 9.9 mL 9.89 mL 9.8 mL 9.8 mL 9.79 mL

9.7 mL 8.9 mL 8.89 mL 8.8 mL

Ibuprofen 0 mL 0 mL 0 mL 0.1 mL 0.1 mL 0.1 mL 1.0 mL 1.0 mL 1.0 mL

Benadryl 0 mL 0.01 mL 0.1 mL 0 mL 0.01 mL

0.1 mL 0 mL 0.01 mL 0.1 mL

Total 10 mL 10 mL 10 mL 10 mL 10 mL 10 mL 10 mL 10 mL 10 mL

100 µL of cell culture was then added to each solution, yielding a final volume of 10 mL and a cell density of approximately 103 cells/mL.

The solutions were vortexed and allowed to sit at room temperature for 15 minutes.

After vortexing to evenly suspend the cells, 100 µL aliquots were removed from the tubes and spread on LB agar plates.

The plates were incubated at 37°C for 48 hours.

The resulting colonies were counted visually. Each colony was assumed to have arisen from one cell.

Procedure

Abbreviation for analysis of variance Statistical test comparing variation within

and between experimental groups If the P- value is lower than the alpha value

(.05), then the result is significant (a result of the variable influence)

Two factor: verifies interaction between two variables.

ANOVA

Staph Survivorship

P-Value: .0508

Concentration (Ibuprofen, Benadryl)

Nu

mb

er o

f Colo

nie

s

E. Coli Survivorship

Concentration (Ibuprofen, Benadryl)

Nu

mb

er o

f Colo

nie

s

The presence of Ibuprofen, Benadryl, and a combination of the two drugs did not significantly change the number of Staphylococcus epidermidis or E. Coli.

The null hypothesis can be accepted. The alternative hypothesis can be rejected

Conclusion

Extensions Limitations

Try prescription medication combination.

Test synergy effects of more harmful drugs.

Use human cells rather than bacteria.

These bacteria were only models for the human cell.

Plates were not plated simultaneously.

http://www.drugs.com/pro/ibuprofen-drug-facts.html

http://textbookofbacteriology.net/staph.html http://www.bacterio.cict.fr/e/escherichia.html http://www.textbookofbacteriology.net/

e.coli.html http://www.drugs.com/mtm/benadryl.html http://www.mayoclinic.com/health/

endocarditis/DS00409

Works Cited