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Caenorhabditis elegans (C. elegans) Kathy Szeniawski Clayton State University Spring 2008

Caenorhabditis elegans (C. elegans) Kathy Szeniawski Clayton State University Spring 2008. Alternate Food Sources Population Growth & Behaviors. Non Parasitic “Bacteria Eating” Nematode 959 Somatic Cells Hermaphrodites and Males Egg/Embryonic Stage L1-L4 molts ? Dauer Stage (Hibernation)

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Caenorhabditis elegans (C. elegans) Kathy Szeniawski Clayton State University Spring 2008

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  1. Caenorhabditis elegans(C. elegans)Kathy SzeniawskiClayton State UniversitySpring 2008 Alternate Food Sources Population Growth & Behaviors

  2. Non Parasitic “Bacteria Eating” Nematode 959 Somatic Cells Hermaphrodites and Males Egg/Embryonic Stage L1-L4 molts ? Dauer Stage (Hibernation) Adult C. elegans Kathy Szeniawski

  3. 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

  4. Why C. elegans? • Easy/Inexpensive To Maintain • Lives in a Petri Dish • Can be easily mutated • 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

  5. 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

  6. C. elegans Worm Farming • Prepare NGM agar plate • Grow an OP-50 E. colilawn • Chunk transfer or Single worm transfer • Grow at room temp • Extract eggs from Adult • Worms can be frozen KathySzeniawski Kathy Szeniawski Kathy Szeniawski

  7. Contamination • Maintain Sterile Techniques Kathy Szeniawski Kathy Szeniawski

  8. 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

  9. What Will the Worms Eat? • OP-50 Bacteria • Fruit Snacks • Tofu • Hoodia chews • Trail Mix (Assorted dried fruits and nuts) • Spinach • Mushrooms

  10. Project: 10 Plates of Each • Spinach • Mushroom • Tofu • Bacteria

  11. + Swarm +++++ ++++ Population Designation (0-5) Few on Surface Several on Surface

  12. ResultsMy Hypothesis is Supported All Populations Survived

  13. Results

  14. 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 Bacteria and Spinach do not support population growth as well as Tofu and Mushrooms

  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

  16. Egg Mounds, Worm Balls & Vortexes

  17. 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

  18. 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

  19. Thank-You • Dr. Francine Norflus, Research Mentor • Dr. Michelle Furlong • Larry and MaryEllen Szeniawski • Kheem Tan • Clayton State University Faculty and Staff

  20. 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.

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