The Behavioral Effects of Aspartame Exposure to Black Planaria (Dugesia dorotocephala) http://s399.photobucket.com/user/chelseybrownfield/media/Lab%2022%20Specimesns/151.jpg.html Jessica L. Pratt Department of Biological Sciences, York College of Pennsylvania Abstract Pilot Study 10 planaria Control No aspartame Objective To determine if there are behavioral and mobility changes after periodic exposure at a low and high aspartame concentration. Hypotheses H1: There are changes in behavior and mobility of black planaria when exposed to aspartame. H2: Changes in behavior and mobility are more frequent as aspartame concentrations increase. 10 planaria Low concentration High concentration 0.001 M 0.01 M Placed in recording petri dish & video recorded planaria for 8 minutes Behavior Time Subjects Interaction Head Bop 0.0754 0.0321* 0.9978 Squirming 0.0176* 0.8850 0.0036** Twitching Head Swing 0.0001*** 0.0082** 0.7247 0.0256* 0.1792 0.1366 Discussion http://www.mindsetsonline.co.uk/Catalogue/ProductDetail/petri-dish?productID=c52d0927-e398-4efc-906e74c0ca95d5bf&catalogueLevelItemID=09ce19f0-e14f-4f2f-99f3-a017466a71e6 http://www.carolina.com/lab-dishes/stender-dish-stacking-type-37-x-25mm/741012.pr;jsessionid=DED2236040ED1740770F47D6EBA4AC27.stageworker2?catId=10261&mCat=&sCat=&ssCat=&questio n= Behavior P < 0.05* P < 0.01** P < 0.001*** Placed in dilution dish • Planaria are freshwater flatworms which are used as an animal model in developmental biology and neuropharmacology research (Raffa and Desai 2005, Pagán et al. 2009). Planaria are known to respond behaviorally to various stimuli such as cocaine, amphetamines, and opiates (Pagán et al. 2009). • There is debate if aspartame is a safe product although it has been approved by the FDA because of its metabolic components. Some of aspartame’s metabolic components, however, can cause changes in the production of neurotransmitters, or when broken down can be very cytotoxic. There is an ongoing debate among researchers whether aspartame causes developmental, behavioral, and neurological effects (Humphries et al. 2007). 10 planaria Exposed for 120 minutes Introduction • Aspartame is a popular artificial sweetener which is used in a large assortment of products. The presence of aspartame in products that are used on a daily basis is growing and not restricted to only sugarfree or unsweetened products. Table 1. Two-way ANOVA summary. Methods Aspartame is a popular artificial sweetener used in a large array of products. There is an ongoing debate among researchers whether aspartame causes developmental, behavioral, and neurological effects. This study investigated whether changes in behavior and mobility of the black planaria (Dugesia dorotocephala) occurred after periodic exposures to low (0.001 M) and high (0.01 M) aspartame concentrations. The control, low, and high concentration groups were kept in separate glass dishes and given fresh water every 3 days after a feeding. Exposures to aspartame or control water occurred in microcentrifuge tubes for 2 hours. This protocol proceeded for 29 days where single-blind observations were recorded every other day. The behaviors searched for were head bopping, head swinging, twitching, cork screwing, squirming, and inch worming. As treatment to aspartame occurred over time a decrease in mobility was observed in planaria. A decrease in mobility was also observed in the control group which gives reason to believe an outside factor could be responsible. Treatment groups differed significantly in the number of head bops and head swings observed. Squirming behavior amounts differed significantly when aspartame concentrations increased. The amount of time the planaria were treated significantly affected the amount of squirming, twitching, and head swinging observed. The amount of time the planaria were treated on the first day against the last significantly affected the amounts of head swings and squirms observed. There were changes in behavior and mobility of black planaria when exposed to aspartame. Changes in behavior and mobility were not more frequent as aspartame concentrations increased. Chemdraw Mobility • As treatment to aspartame occurred over time a decrease in mobility was observed in planaria, along with the control group. The outside factor hypothesized to be responsible was contaminated river water from the collection site at Tyler Run Creek (Figure 1). • Treatment groups differed significantly in the number of head bops and head swings (Table 1). • Squirming behavior amounts differed significantly when aspartame concentrations increased (Table 1, Figure 4). • The amount of time the planaria were treated significantly affected the amount of squirming, twitching, and head swinging (Table 1). • The amount of time the planaria were treated on the first day vs the last day, significantly affected the amounts of head swings and squirms observed (Figure 2, Figure 4). Conclusions Head Bopping Head Swinging Twitching Cork Screwing Results Squirming Inch Worming • There were changes in behavior and mobility of black planaria when exposed to aspartame. • Changes in behavior and mobility were not more frequent as aspartame concentrations increased. • The decrease in mobility within the control group gives reason to believe an outside factor was responsible. Literature Cited • Humphries, P.P., Pretorius, E.E., and Naudé, H.H. (2008). Direct and indirect cellular effects of aspartame on the brain. European Journal Of Clinical Nutrition  62(4): 451-462. • Lavin, J., French, S., & Read, N. (1997). The effect of sucrose- and aspartamesweetened drinks on energy intake, hunger and food choice of female, moderately restrained eaters. International Journal Of Obesity & Related Metabolic Disorders  21(1): 37. • Pagán, R.O., Coudron, T., and Kaneria, T. (2009). The Flatworm Planaria as a Toxicology and Behavioral Pharmacology Animal Model in Undergraduate Research Experiences. The Journal of Undergraduate Neuroscience Education [serial online] 7(2): A48-A52. • Pagán R.O., Rowlands, L.A., and Urban R.K. 2006. Toxicity and behavioral effects of dimethylsulfoxide in planaria. Neuroscience Letters. [0304-3940] 407(3): 274-278. • Raffa, B.R. and Desai, P. 2005. Description and quantification of cocaine withdrawal signs in Planaria. Brain Research [0006-8993] 1032(1-2): 200-202.0 Acknowledgements I would like to thank York College of Pennsylvania for funding my research. I would like to thank Dr. Rehnberg for his guidance throughout the entirety of my research. I would like to thank Dr. Hagerty for her advice in the analysis of my data. I would like to thank Joan Carpenter for her continuous help in ensuring lab access throughout the summer months. Lastly, I would like to thank TJ Steckbeck, Masi Luttner, and Emily Springer for assisting me during the course of my research.