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The Effects of Different Strains of Pseudomonas aeruginosa on Drosophila melanogastar Need http://knol.google.com/k/-/-/UtI7gr91/rIGkBA/Slide4.jpg Knowledge Base Characteristics of pseudomonas aeruginosa • Tolerant to a wide variety of physical conditions http://cmbi.bjmu.edu.cn/news/0310/21.htm Knowledge Base Immune Response of Drosophila melanogastar • Innate immunity is activated when microorganisms succeed in entering the body cavity of the fly. • Invading microorganisms are killed by the combined action of cellular and humoral processes • phagocytosed by specialized blood cells, surrounded by toxic melanin, or lysed by antibacterial peptides secreted into the hemolymph by fat body cells Knowledge Base Quorum Sensing • enables bacteria to co-ordinate their behavior http://www.quiestech.com/content/science/quorum-sensing_more.asp Knowledge Base Quorum Sensing • Signal molecule used by pseudomonas aeruginosa http://www.nottingham.ac.uk/quorum/what.htm Literature Review • Stoltz, et al. (2008) – pricking the fly’s abdomen with a needle previously dipped in a P. aeruginosa suspension – PON1 transgenic flies are protected from lethality due to P. aeruginosa infection Literature Review • D’Argenio, et al (2000) – PAO1, the best-characterized strain of P. aeruginosa was used – Adult female flies 2 to 4 days old were pricked in the dorsal thorax with a 25-gauge needle dipped directly into the PAO1 culture Time course of infection of wild-type flies with PAO1 ( ), a pilD mutant ( ), and a chpA mutant ( ). Bacteria were grown as described for PAO1 and introduced into flies by pricking with a syringe needle. The chpA37 mutant was used for this and subsequent experiments. The values plotted are the averages of five replicate experiments, each with 10 flies, and the standard error of the mean is shown for each point. Purpose The purpose of this experiment is to see if Drosophila melanogastar can be protected from the lethal strain of pseudomonas aeruginosa by being injected with the non-lethal strain first Hypothesis Null hypothesis- The non-lethal strain of pseudomonas aeruginosa will have no effect on the protection of the flies from the lethal strain Alternate hypothesis- The non-lethal strain will have an effect on the protection of the flies from the lethal strain Methodology Drosophila melanogastar N=60 Flies put under anesthesia Injected with non lethal strain Injected with lethal strain N=30 N=30 Injected with lethal strain Amount of living flies are counted Statistical analysis by ANOVA with p<.05 Protocols For Measurement • The amount of dead or living flies will be counted manually Do-Ability • Flies are easily obtained, cheap, and easy to keep alive • Bacteria can be kept alive using culturing techniques from articles that were read Budget Vendor Catalogue # Page Carolina Biological Frey Scientific Carolina Biological 025 97 Item Item # Quantity Unit $ Total $ FlyNap 173010 5 $12.50 $62.50 Drosophila Culture 56348501 6 1 $13.50 $13.50 Pseudomo nas aeruginosa MicroKwik Culture®, Vial 155250A 1 $12.00 $12.00 $88.00 Bibliography