By: Patrick Leech Pittsburgh Central Catholic PJAS 2010 Grade 10

Entry into the water system Many pharmaceuticals enter the water through human excretion. Direct disposal through household plumbing Water treatment plants only remove about 60% of the drugs.

Benadryl Used as an antihistamine Works by blocking the effect of histamine receptor sites Active Ingredient: Diphenhydramine hydrochloride Does not have FDA safe to flush label on it Trace amounts have been found in drinking water in parts of the country Over 25 common over the counter drugs contain the same active ingredient of Benadryl (Diphenhydramine hydrochloride)

Ibuprofen A non-steroidal anti-inflammatory drug Used for relief of symptoms of arthritis, fever, and joint pains 3 rd most consumed drug in the world About 30% of the amount of Ibuprofen bought enters the water Studies have shown that even low concentrations have been found to affect the way fish spawn. As Ibuprofen decomposes, it becomes more toxic. Does not have FDA approved “safe to flush” label on it

Synergy? The two medicines Benadryl and Ibuprofen mixed together

Algae Large diverse group of simple and usually autotrophic organisms Base of aquatic food chain Used as a bio-indicator for aquatic environments

Euglena Generally cylindrical in shape with many flagella Common algal like protist A partial heterotroph can attain food through endocytosis and photosynthesis Nutrient-rich freshwater or in sewage systems Capable of survival in both salt and water environments Frequently employed as an experimental model

Purpose To discern the effect(s) of various concentrations of Ibuprofen, Benadryl, and a combination of both on the growth/survivorship of Euglena

Hypothesis Null: The presence of Benadryl, Ibuprofen, and a combination of both will NOT have a significant effect on algal growth. Alternative: The presence of Benadryl, Ibuprofen, and a combination of both WILL have a significant on algal growth.

Materials 10% Benadryl solution (10 grams Benadryl per 100mL Spring water) 10% Ibuprofen solution (10 grams Benadryl per 100mL Spring water) Euglena gracilis Soil water (sterile) Spring water 90 test tubes (13 x 100 mm borosilicate culture tubes) Pipettes (macro + micro) Pipette tips Science Kit Educator spectrophotometer Test tube rack

Procedure 1. A desk lamp was placed in experimental site a) approximately 45 centimeters away from the test tube racks b) Kept on a 12 hour on 12 hour off cycle c) Kept at 22 degrees celsius throughout the duration of the study 2. Created a 10% stock solution of Ibuprofen, Benadryl, and a combination of both 3. Created the concentrations of 0%, 0.01%, and 0.1% for Ibuprofen, Benadryl, and the combination a)There were ten replicates per concentration 4. Readings were taken using a spectrophotometer set with a wavelength of 430 nm once a day on days 1-10

Test Tube Ingredients Concentration0%0.01%0.1% Algae2mL Distilled Water3mL2.995mL2.95mL 10% stock solution0mL0.005mL0.05mL Total5mL

Benadryl Influence on Euglena Population Growth

Ibuprofen Influence on Euglena Population

Synergy Influence on Euglena Population

Results Is algal growth affected by the presence of Benadryl or Ibuprofen in its environment? Ibuprofen? P value NOT SIGNIFICANT Benadryl? P value NOT SIGNIFICANT Synergy P value NOT SIGNIFCANT Alpha =.05

Conclusions The null hypothesis was supported because any variation that occurred was insignificant according to the ANOVA statistical analysis After reviewing the statistical analysis the P value for all three variables were greater than the standard cut off of.05 Because the P-value was greater than.05 on all concentrations a Pair-wise test (Dunnett’s Test) was not needed

Limitations and Extensions Limitations Euglena not fresh (Used 4 days after arrival) Extensions Use more diverse groups of algae Use a higher concentration of the drugs Use different kinds of drugs both over the counter and prescription Use hemocytometer to obtain more accurate results

References =usccc Science-Examiner~y2010m1d19-Benadryl-Motrin-and- Rolaids-added-to-recall-of-Tylenolmakers-products