1. Effects of Caffeine and Ibuprofen on the Growth of Arab Kyle Butzine, Jasmine Crafton, and Dr. Catherine Chan University of Wisconsin – Whitewater, Department of Biological Sciences and Chemistry Introduction Pharmaceuticals and personal care products (PPCPs) are often released into the environment, even after wastewater treatment (e.g., application of biosolids and wastewater runoff) Two commonly consumed pharmaceuticals are caffeine and ibuprofen. The effects of caffeine and ibuprofen in the environment have not been well studied. We test the effects of caffeine and ibuprofen on various growth aspects of a model land plant, Arabidopsis thaliana, at different life stages. The goal is to examine how these drugs affect plant growth and their effective concentrations. Ibuprofen Methods Germination Experiment Grow A. thaliana seeds in standard nutrient agar media enriched with various concentrations of caffeine and ibuprofen. Early Growth Experiment Grow A. thaliana seeds in regular nutrient media. Transplant seedlings into caffeine- and ibuprofen-containing media when plants have reached the 4 leaf growth stage.. Compare primary root length of plants transplanted into caffeine and ibuprofen media to plants grown in regular nutrient media. Record the length of time it takes for roots, stem, 2 leaves, 4 leaves, and 6 leaves to emerge. Later Stage Growth Experiment Continuous Exposure Grow A. thaliana in regular nutrient media until the 4 leaf developmental stage is reached. Transfer plants into soil and divide plants into control and experimental group. Water control group with water and the experimental with water supplemented with 20 mM of calcium chloride. Water every 3 days. Direct Application Separate flower clusters and leaves into control and experimental groups. Apply a set volume of ibuprofen or caffeine at different concentrations on control and experimental flowers and leaves. Repeat the same with solvent control. Repeat every 3 days and record differences between the control and experimental groups. Acknowledgements Merck/ AAAS Summer Undergraduate Research Program WiscAMP College of Letters and Sciences University of Wisconsin Whitewater Undergraduate Research Program Figure 1: Average % of seeds that germinated in different concentrations of ibuprofen within 317 hours (about 13 days). Without ibuprofen, 89% of seeds germinated. Error bars are calculated based on 95% confidence. As concentration of ibuprofen increases, a statistically significant percentage of seeds that germinate decreases. Figure 3: Average time for appearance of different plant organs of plants grown in various concentrations of ibuprofen. Plants exposed to 10 ppm of ibuprofen did not grow 4 leaves or 6 leaves within the experimental time frame. Plants exposed to 10 PPM of Ibuprofen show a statistically significant delay of all organs. Figure 2: Average % of seeds that germinated in different concentrations of caffeine within 317 hours (about 13 days). Without caffeine, 78% of seeds germinated. Error bars are based on 95% confidence. No differences in germination between the presence and absence of caffeine or exposure to different concentrations of caffeine are observed. Figure 6: Average amount of primary root growth after plants were exposed to various caffeine concentrations during the 4 leaf stage for 24 hours. Statistically significant differences are seen at 100 ppm caffeine. Preliminary data suggests that 2 leaf stage is unaffected by concentrations up to 1 ppm and 6 leaf is unaffected up to 100 ppm caffeine. Germination Results Caffeine Figure 4: Average time for appearance of different plant organs of plants grown in various concentrations of caffeine. Caffeine appears to not significantly delay organ emergence. Figure 5: Average amount of primary root growth after plants were exposed to various ibuprofen concentrations during the 4 leaf stage for 24 hours. Plants exposed to 10 ppm and 100 ppm of ibuprofen have a significant decrease in root length. Preliminary data suggests that 2 and 4 leaf stages plants have similar responses toward ibuprofen. Figure 3 Figure 5 Figure 7: Plants grown in 10 ppm and 100 ppm ibuprofen showed lateral root growth, as oppose to the normal downward growth aligned with the gravity vector. No Ibuprofen10 ppm Ibuprofen Figure 1 Figure 3 Figure 5 Figure 4 Figure 6 Caffeine Early Growth Results Ibuprofen Caffeine