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Caenorhabditis elegans (C. elegans) Kathy Szeniawski Clayton State University Spring 2008 Alternate Food Sources Population Growth & Behaviors C. elegans Non Parasitic “Bacteria Eating” Nematode 959 Somatic Cells Hermaphrodites and Males Egg/Embryonic Stage L1-L4 molts ? Dauer Stage (Hibernation) Adult Kathy Szeniawski C. elegans Natural Habitat Nutrient and Microorganism rich organic material Cultures obtained from compost/garden soil, rotting fruit and mushroom beds Associated with millipedes, insects, snails and slugs Photoblog.com Queensbury net Paintworks Why C. elegans? Feeds on OP-50 E. coli Bacteria in the lab Short Life Cycle/Can produce several generations in a short period of time Perfect for Genetic Research Kathy Szeniawski Easy/Inexpensive To Maintain Lives in a Petri Dish Can be easily mutated OP-50 E. coli Grow OP-50 E. coli at 37 degrees C overnight in 2X YTB (Yeast Tryptone Broth) Pipette onto NGM agar plates Grow your “Bacterial Lawn” for 2 days Kathy Szeniawski Kathy Szeniawski Kathy Szeniawski C. elegans Worm Farming Prepare NGM agar plate Grow an OP-50 E. coli lawn Chunk transfer or Single worm transfer Grow at room temp Extract eggs from Adult Worms can be frozen Kathy Szeniawski Kathy Szeniawski Kathy Szeniawski Contamination Maintain Sterile Techniques Kathy Szeniawski Kathy Szeniawski Worm Research Research papers on “Different food source” experiments used different strains of bacteria to feed to the worms. I hypothesized that wild type worms in their natural environment consumed food items since the lab worms that were collected were located in compost piles, mushroom beds and rotten fruit. N2 wild type (normal) worms used Transfer worms onto plates containing food products Observe and document population growth and behavior What Will the Worms Eat? OP-50 Bacteria Fruit Snacks Tofu Hoodia chews Trail Mix (Assorted dried fruits and nuts) Spinach Mushrooms Project: 10 Plates of Each Spinach Mushroom Tofu Bacteria Population Designation (0-5) + Swarm +++++ Few on Surface ++++ Several on Surface Results My Hypothesis is Supported Population 5 Size 4 Bacteria 3 Tofu 2 Spinach Mushroom 1 0 1 2 3 6 7 8 9 10 13 15 Days All Populations Survived Results Population 5 Size 4 Bacteria 3 Tofu 2 Spinach Mushroom 1 0 1 2 3 6 7 8 Days 9 10 13 15 Spinach 5 T test Day 8 p values B/T= 9.915x10-6 B/S = 0.017 B/M = 7.77x10-6 T test Day 15 p values B/T = 0.020 B/S = 1 B/M = 1.62x10-6 Size 4 3 Bacteria 2 Spinach 1 0 1 2 3 6 7 8 9 10 13 15 Days Bacteria and Spinach do not support population growth as well as Tofu and Mushrooms Mushrooms 5 5 4 4 3 Bacteria 2 Tofu Size Size Tofu 3 Bacteria 2 Mushroom 1 1 0 0 1 2 3 6 7 8 Days 9 10 13 15 1 2 3 6 7 8 Days 9 10 13 15 Unusual Behaviors Observed Worms eating food products Egg laying balls of Hermaphrodites Egg mounds visible to the naked eye Larval worm balls Larval worms participating in Cannibalism Larval worms killing and eating dust mites Vortexes in food Egg Mounds, Worm Balls & Vortexes Future Research Repeat experiment with controlled seeding Use equal masses of food products on all plates Collect statistical data every day – Population size – Amounts of Large worms vs. Small worms Observe behavior Conclusion Different food sources CAN sustain C. elegans Cannibalistic behavior observed several times; Only larval worms observed participating in cannibalism Larval worms observed killing/eating dust mites Overall Population size quickly spikes, then tapers off until only a few Dauer worms exist Worm size in population tapers off to all small worms at the end of the growth curve Thank-You Dr. Francine Norflus, Research Mentor Dr. Michelle Furlong Larry and MaryEllen Szeniawski Kheem Tan Clayton State University Faculty and Staff References Altn, Z.F. and Hall, D.H. 2005. Handbook of C. Elegans Anatomy. Introduction to C. Elegans Anatomy. In WormAtlas. http://www.wormatlas.ord/handbook.contents.htm Kiontke, Karin. and Sudhaus, Walter. Ecology of Caenorhabditis species. (January 09, 2006), Wormbook, ed. The C. Elegans research community, Wormbook, doi/10.1895/wormbook.1.37.1, http://www.wormbook.org Brenner, Sydney. The worm goes to school. 2003. Institute of Molecular and Cell Biology and National Institute of Education Barriere, Antoine. And Felix, Marie-Anne. Isolation of C. Elegans and related nematodes. (July 17, 2006), Wormbook, ed. The C. Elegans research community, WormBook, doi/10.1895/wormbool.1.115.1 http://www.wormbook.org Shtonda, Boris and Avery, Leon. Dietary choice behavior in Caenorhabditis elegans. 2006 The Journal of Experimental Biology 209, 89-102.