2. Need Figure 1: This pie chart shows the percentage of deaths related from bacterial infection present in major countries in January 2004. The U.S has the greatest percentage of 44.6% http://www.nationmaster.com/red/pie/mor_bac_inf_of_uns_sit- mortality-bacterial-infection-unspecified-site

3. Knowledge Base Figure 2: the physiology of a mollusk and the main parts the of organism http://www.bio.miami.edu/dana/106/106F06_12.html

4. Knowledge Base http://www.bodyfasting.com/juice-detox-fasting.html Figure 3: Antioxidant replacing free radicals

5. Knowledge Base Figure 4: Nutrition facts of Spirulina http://www.spirulina.com/ nutrition_tech_info.html

6. Knowledge Base Figure 5: Nutrition facts of Spirulina http://www.spirulina.com/ nutrition_tech_info.html

7. Literature Review (Ottaviani, 2004) Figure 6: the evolutionary path of immune- neuroendocrine response between mammals and invertebrates Mollusks are ideal model organisms for immune response due to their similarities in immune and nuroendocrine systems

8. Literature Review Figure 7: schistosome Trichobilharzia ocellata that was exposed to one group of the pond snails When the snails that were exposed and unexposed to Trichobilharzia ocellata were compared their immune responses to an infection of Staphylococcus saprophyticus bacteria differed. (Van der Knaap, et al, 1986)

9. Literature Review (Van der Knaap, et al, 2004) the humoral defense activities of a Lymnaea stagnalis agglutination, oposonisation, and inhibition of bacterial growth.

10. Literature Review (Brambilla, et al, 2008) Antioxidants are used on a large scale to try to maintain health Antioxidants may be used to prevent free radical related health disorders

11. Purpose/Hypothesis Purpose: The purpose of this experiment is to show the effect that antioxidants have on immune responses to bacterial infections Null Hypothesis: There will be no significant difference between the immune response to the e.coli when the pond snail is either exposed or not exposed to antioxidants Alternate Hypothesis: There will be a significant difference between the immune response to the e.coli when the pond snail is exposed to the antioxidant and when it is not exposed.

12. Methodology Pond Snails (24) Control- infected with e.coli fed with chorella (no antioxidants) Sample Size: 12 Variable – infected with e.coli fed with Spirulina (antioxidants) Sample Size: 12 Measurement Immune response compared to variable and length of infection in pond snails Statistical Analysis Measurement Immune response compared to variable and length of infection in pond snails

13. Protocols for Measurement Differences in the reaction to the e.coli between the snails exposed to antioxidants and the unexposed snails Survival Rate of Snails

14. Do-Ability Freshwater pond snails: easy to keep alive and maintain No expensive equipment needed Some equipment needed is available in the lab Qualitive data is obtained by observation of the pond snails immune response to the bacterial infection

15. Budget vendorcatolog #pagedescriptionquantityunit pricetotal price carolina141214 pond snails1$17.95 wards85 V 1860 Escherichia coli1$15.50 wards86 V 0700 Spirulina1$6.75 carolina162101 Elodea1$7.25 carolina671226 Aquarium Tank, Glass, 2 1/2 gal2$17.16$34.32 carolina671227 Tank Cover, Glass, for 2 1/2 gal2$8.27$16.54 total$97.81

16. Bibliography " Bacterial infection of unspecified site by country. Definition, graph and map." NationMaster - World Statistics, Country Comparisons. 02 June 2009. Best, Ben. "GENERAL ANTIOXIDANT ACTIONS." BEN BEST's HOME PAGE. 02 June 2009. "BIL 106 - Lecture 12." University of Miami Department of Biology. 04 June 2009. Brambilla, Daria, Cesare Mancuso, Mariagrazia R. Scuderi, Paolo Bosco, Giuseppina Cantarella, Laurence Lempereur, Giulia D. Benedetto, Salvatore Pezzino, and Renato Bernardini. "The role of antioxidant supplement in immune system, neoplastic, and neurodegenerative disorders: a point of view for an assessment of the risk/benefit profile." Nutrition Journal 7 (2008). BioMed Central. 30 Sept. 2008. 10 June 2009. Lockyer, Anne E., Jennifer N. Spinks, Anthony J. Walker, Richard A. Kane, Leslie R. Noble, David Rollinson, Emmanuel Dias-Neto, and Catherine S. Jones. "Biomphalaria glabrata transcriptome: Identification of cell-signalling, transcriptional control and immune-related genes from open reading frame expressed sequence tags (ORESTES)." Development and Comparitive Immunology 8 (2007): 763-82. PubMed Central. 2007. 10 June 2009. Maule, Aaron. Parasitic flatworms molecular biology, biochemistry, immunology and physiology. Cambridge, MA: CABI Pub., 2005. Pacheco-Lopez, Gustavo, Maj-Britt Niemi, Harald Engler, and Manfred Schedlowski. "Neuro-Immune Associative Learning." Neural Plasticity and Memory (2007). CRC Press. 2007. 10 June 2009. Ottaviani, Enzo. "The mollusc as a suitable model for mammalian immune-neuroendocrine." Http://www.isj.unimo.it/articoli/ISJ001.pdf 1 (2004): 2-4. Visions and Perspectives. 2004. 13 May 2009. Pacheco-López, Gustavo, Maj-Britt Niemi, Harald Engler, and Manfred Schedlowski. "Neuro-Immune Associative Learning." Neural Plasticity and Memory. Frontiers in Neuroscience. 2007. CRC Press. 5 June 2009. "Spirulina Earthrise - Nutrition and Technical Information." Spirulina Earthrise - Health Benefits of Spirulina, The World's Healthiest Superfood,. 04 June 2009. Van der Knaap, W. P. W., E. A. Meuleman, and T. Sminia. "Alterations in the internal defence system of the pond snail Lynmaea stagnalis induced by infection with the schistosome Trichobilharzia ocellata." Parasitology Research 73 (1987): 57-65. SpringerLink. 12 Dec. 2004. 9 June 2009. Van der Knaap, W.P.W, C. M. Adema, and T. Sminia. "Invertebrate blood cells: Morphological and functional aspcts of the haemocytes in teh pond snail Lymnaea stagnalis." Comparative Haematology International 3 (1993): 20-26. SpringerLink. 11 Dec. 2004. 9 June 2009.