caenorhabditis elegans (c. elegans) kathy szeniawski clayton state university spring 2008
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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) - PowerPoint PPT PresentationTRANSCRIPT
Caenorhabditis elegansCaenorhabditis elegans(C. elegans)(C. elegans)
Kathy SzeniawskiKathy SzeniawskiClayton State UniversityClayton State University
Spring 2008Spring 2008
Alternate Food SourcesAlternate Food Sources
Population GrowthPopulation Growth
& Behaviors& Behaviors
C. elegansC. elegans Non ParasiticNon Parasitic ““Bacteria Bacteria
Eating” Eating” NematodeNematode
959 Somatic 959 Somatic CellsCells
Hermaphrodites Hermaphrodites and Malesand Males
Egg/Embryonic Egg/Embryonic StageStage
L1-L4 moltsL1-L4 molts ? Dauer Stage ? Dauer Stage
(Hibernation)(Hibernation) AdultAdult
Kathy Szeniawski
C. elegansC. elegans Natural Natural HabitatHabitat
Nutrient and Nutrient and Microorganism rich Microorganism rich organic materialorganic material
Cultures obtained Cultures obtained from from compost/garden compost/garden soil, rotting fruit soil, rotting fruit and mushroom and mushroom bedsbeds
Associated with Associated with millipedes, insects, millipedes, insects, snails and slugssnails and slugs
Photoblog.com Queensbury net
Paintworks
Why Why C. elegansC. elegans?? Easy/Inexpensive To Easy/Inexpensive To
MaintainMaintain Lives in a Petri DishLives in a Petri Dish Can be easily mutatedCan be easily mutated Feeds on OP-50 Feeds on OP-50 E. E.
colicoli Bacteria in the Bacteria in the lablab
Short Life Cycle/Can Short Life Cycle/Can produce several produce several generations in a short generations in a short period of timeperiod of time
Perfect for Genetic Perfect for Genetic ResearchResearch
Kathy Szeniawski
OP-50 OP-50 E. coliE. coli Grow OP-50 Grow OP-50 E. E.
colicoli at 37 degrees at 37 degrees C overnight in 2X C overnight in 2X YTB (Yeast Tryptone YTB (Yeast Tryptone Broth)Broth)
Pipette onto NGM Pipette onto NGM agar platesagar plates
Grow your Grow your “Bacterial Lawn” for “Bacterial Lawn” for 2 days2 days
Kathy Szeniawski
Kathy Szeniawski
Kathy Szeniawski
C. elegansC. elegans Worm Worm FarmingFarming
Prepare NGM agar Prepare NGM agar plateplate
Grow an OP-50 Grow an OP-50 E. E. colicoli lawn lawn
Chunk transfer or Chunk transfer or Single worm transferSingle worm transfer
Grow at room tempGrow at room temp Extract eggs from Extract eggs from
AdultAdult Worms can be frozen Worms can be frozen
Kathy Szeniawski
Kathy Szeniawski
Kathy Szeniawski
ContaminationContamination
Maintain Sterile TechniquesMaintain Sterile Techniques
Kathy Szeniawski
Kathy Szeniawski
Worm ResearchWorm Research Research papers on “Different food source” Research papers on “Different food source”
experiments used different strains of experiments used different strains of bacteriabacteria to to feed to the worms. feed to the worms.
I hypothesized that wild type worms in their I hypothesized that wild type worms in their natural environment consumed food items since natural environment consumed food items since the lab worms that were collected were located in the lab worms that were collected were located in compost piles, mushroom beds and rotten fruit. compost piles, mushroom beds and rotten fruit.
N2 wild type (normal) worms usedN2 wild type (normal) worms used Transfer worms onto plates containing food Transfer worms onto plates containing food
productsproducts Observe and document population growth and Observe and document population growth and
behaviorbehavior
What Will the Worms Eat?What Will the Worms Eat?
OP-50 BacteriaOP-50 Bacteria Fruit SnacksFruit Snacks TofuTofu Hoodia chewsHoodia chews Trail Mix (Assorted Trail Mix (Assorted
dried fruits and dried fruits and nuts)nuts)
SpinachSpinach MushroomsMushrooms
Project: 10 Plates of EachProject: 10 Plates of Each
SpinachSpinach MushroomMushroom Tofu Tofu BacteriaBacteria
Population Designation (0-5)Population Designation (0-5)
++
Swarm +++++Swarm +++++
++++++++Few on Surface Several on Surface
ResultsResultsMy Hypothesis is SupportedMy Hypothesis is Supported
Population
0
1
2
3
4
5
1 2 3 6 7 8 9 10 13 15
Days
Siz
e
Bacteria
Tofu
Spinach
Mushroom
All Populations Survived
ResultsResults
Population
0
1
2
3
4
5
1 2 3 6 7 8 9 10 13 15
Days
Siz
e
Bacteria
Tofu
Spinach
Mushroom
Bacteria and Spinach do not support population growth as well as Tofu and Mushrooms
Tofu
0
1
2
3
4
5
1 2 3 6 7 8 9 10 13 15
Days
Siz
e Bacteria
Tofu
Spinach
0
1
2
3
4
5
1 2 3 6 7 8 9 10 13 15
Days
Siz
e Bacteria
Spinach
Mushrooms
0
1
2
3
4
5
1 2 3 6 7 8 9 10 13 15
Days
Siz
e Bacteria
Mushroom
T test Day 8p valuesB/T= 9.915x10-6
B/S = 0.017B/M = 7.77x10-6
T test Day 15p valuesB/T = 0.020B/S = 1B/M = 1.62x10-6
Unusual Behaviors Unusual Behaviors ObservedObserved
Worms eating food productsWorms eating food products Egg laying balls of HermaphroditesEgg laying balls of Hermaphrodites Egg mounds visible to the naked eyeEgg mounds visible to the naked eye Larval worm ballsLarval worm balls Larval worms participating in CannibalismLarval worms participating in Cannibalism Larval worms killing and eating dust mitesLarval worms killing and eating dust mites Vortexes in foodVortexes in food
Egg Mounds, Worm Balls & Egg Mounds, Worm Balls & VortexesVortexes
Future ResearchFuture Research
Repeat experiment with controlled Repeat experiment with controlled seedingseeding
Use equal masses of food products Use equal masses of food products on all plateson all plates
Collect statistical data every day Collect statistical data every day – Population sizePopulation size– Amounts of Large worms vs. Small Amounts of Large worms vs. Small
wormsworms Observe behaviorObserve behavior
ConclusionConclusion
Different food sources CAN sustain Different food sources CAN sustain C. elegansC. elegans
Cannibalistic behavior observed several times; Cannibalistic behavior observed several times; Only larval worms observed participating in Only larval worms observed participating in cannibalismcannibalism
Larval worms observed killing/eating dust mitesLarval worms observed killing/eating dust mites Overall Population size quickly spikes, then Overall Population size quickly spikes, then
tapers off until only a few Dauer worms existtapers off until only a few Dauer worms exist Worm size in population tapers off to all small Worm size in population tapers off to all small
worms at the end of the growth curveworms at the end of the growth curve
Thank-YouThank-You
Dr. Francine Norflus, Research Dr. Francine Norflus, Research MentorMentor
Dr. Michelle FurlongDr. Michelle Furlong Larry and MaryEllen SzeniawskiLarry and MaryEllen Szeniawski Kheem TanKheem Tan Clayton State University Faculty and Clayton State University Faculty and
StaffStaff
ReferencesReferences Altn, Z.F. and Hall, D.H. 2005. Handbook of Altn, Z.F. and Hall, D.H. 2005. Handbook of C. ElegansC. Elegans
Anatomy. Introduction to C. Elegans Anatomy. In Anatomy. Introduction to C. Elegans Anatomy. In WormAtlas. WormAtlas. http://http://www.wormatlas.ord/handbook.contents.htmwww.wormatlas.ord/handbook.contents.htm
Kiontke, Karin. and Sudhaus, Walter. Ecology of Kiontke, Karin. and Sudhaus, Walter. Ecology of CaenorhabditisCaenorhabditis species. (January 09, 2006), Wormbook, ed. species. (January 09, 2006), Wormbook, ed. The The C. ElegansC. Elegans research community, Wormbook, research community, Wormbook, doi/10.1895/wormbook.1.37.1, doi/10.1895/wormbook.1.37.1, http://http://www.wormbook.orgwww.wormbook.org
Brenner, Sydney. The worm goes to school. 2003. Institute Brenner, Sydney. The worm goes to school. 2003. Institute of Molecular and Cell Biology and National Institute of of Molecular and Cell Biology and National Institute of EducationEducation
Barriere, Antoine. And Felix, Marie-Anne. Isolation of Barriere, Antoine. And Felix, Marie-Anne. Isolation of C. C. ElegansElegans and related nematodes. (July 17, 2006), Wormbook, and related nematodes. (July 17, 2006), Wormbook, ed. The C. Elegans research community, WormBook, ed. The C. Elegans research community, WormBook, doi/10.1895/wormbool.1.115.1 doi/10.1895/wormbool.1.115.1 http://http://www.wormbook.orgwww.wormbook.org
Shtonda, Boris and Avery, Leon. Dietary choice behavior in Shtonda, Boris and Avery, Leon. Dietary choice behavior in Caenorhabditis elegansCaenorhabditis elegans. 2006 The Journal of Experimental . 2006 The Journal of Experimental Biology 209, 89-102. Biology 209, 89-102.