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Quantitative Investigations of Hatching in Brine Shrimp Cysts

Quantitative Investigations of Hatching in Brine Shrimp Cysts. Charlie Drewes Ecology, Evolution & Organismal Biology Iowa State University, Ames, IA. Artemia franciscana North American brine shrimp. Class Branchiopoda. meaning: “gilled feet”. gilled feet. ● crustacean subgroup.

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Quantitative Investigations of Hatching in Brine Shrimp Cysts

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  1. Quantitative Investigations of Hatching in Brine Shrimp Cysts Charlie Drewes Ecology, Evolution & Organismal Biology Iowa State University, Ames, IA

  2. Artemia franciscanaNorth American brine shrimp

  3. Class Branchiopoda meaning: “gilled feet” gilled feet ●crustacean subgroup ●e.g. fairy shrimp, brine shrimp, clam shrimp, tadpole shrimp & daphnids ● almost all freshwater ● use leaf-like appendages to feed, swim & respire ● often produce anabiotic eggs

  4. fertilized eggs in brood sac Artemialife cycle release of encysted gastrula stage embryos = ‘winter eggs’ Adult female

  5. Artemia franciscana(dry, encysted ‘winter eggs’)

  6. ASW Artemialife cycle X winter ‘eggs’ HATCH

  7. Advantages of “stuck-on Artemia” > easy transfer & handling of cysts >quantitate starting # of cysts and # hatched > follow development & hatching of single cysts > view hatching cysts in a single focal plane >high rates of hatching & survival >no aeration required >conservative use of cysts > open-ended investigations (many variables)

  8. ‘Paint’ cysts on double-stick tape with small, soft, dry paintbrush

  9. Method A Method Apaint cysts on sticky circle

  10. Method B plastic petri dish transparency strip with grid10 mm x 10 mm strip of double-stick tape covers grid tape holds strip to dish

  11.  Cysts painted on sticky grid with brush

  12. A1 Hatching of encystedArtemia embryos A1-A2: Encysted gastrulae in ASW and stuck to tapeB: Cyst swells and wall splitsC1-C2: ‘Umbrella’ stage emerges (membrane-bound) D1-D2: Release of free-swimming nauplius larvae C1 A2 C2 B D1 D2

  13. Each day, a pipet is used to remove and count hatched, swimming nauplius larvae

  14. CONTROL 50 0 0 0% 22 22 44% 66% 11 33 6 39 78% 1 40 80%

  15. Typical hatching curve: %HATCH

  16. Examples of experimental variables that may affect hatching curves 1. dry cysts exposed to thermal extremes 2. ambient light effects during hydration 3. temperature effects during hydration 4. salinity effects 5. second hydration 6. oxygenation effects during hydration 7. environmental toxicant effects 8. irradiation effects on dry or hydrating cysts

  17. Measurements & End-points * Hatching % *hatching time-coursedevelopmental anomalieslarval motility molting & growth

  18. 'N Cyst A sea monkey's no monkey or chimp. It's crustacean - better known as brine shrimp.This genus Artemia Can deal with extremia.As a cyst stage, its hardly a wimp.

  19. 'N Cyst A sea monkey's no monkey or chimp. It's crustacean - better known as brine shrimp.This genus Artemia Can deal with extremia.As a cyst stage, its hardly a wimp.

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