Type of Document Master's Thesis Author Herbst, Eric Christopher URN etd-0711102-135713 Title Induction of Tetraploidy in Zebrafish Danio rerio and Nile Tilapia Oreochromis niloticus Degree Master of Science (M.S.) Department Forestry, Wildlife, and Fisheries Advisory Committee
Advisor Name Title Terrence R. Tiersch Committee Chair C. Greg Lutz Committee Member Robert P. Romaire Committee Member Keywords
- egg hatching system
- nile tilapia
- single female spamning
Date of Defense 2002-06-05 Availability unrestricted AbstractThe goals of this thesis were to: 1) induce tetraploidy in zebrafish; 2) characterize single-female spawning in Nile tilapia for egg collection; 3) design and construct hatching systems for tilapia eggs; 4) induce tetraploidy in Nile tilapia.
Tetraploidy was induced in zebrafish Danio rerio, by applying a 41 C heat shock to eggs for 2 min at 11, 13, 15, 17, or 20 min after fertilization (AF). A trend of increasing percent tetraploid yield occurred the later the shocks were applied. Shocks applied at 11 min AF yielded 1% tetraploidy and shocks applied at 20 min AF yielded 10%. There were no significant differences (P = 0.20) in induction among treatments. This is the first report on induction of tetraploidy in zebrafish.
Stocking of single-female Nile tilapia Oreochromis niloticus in ten 80-L aquaria yielded a reliable supply of eggs for polyploidy research conducted in this study. In one year, 72 spawns were recorded, yielding ~90,000 eggs. Peak spawning occurred in summer.
The two egg hatching systems for artificial incubation of tilapia eggs proved to be effective, with 75% of control fish surviving to hatch. Methylene blue and ultraviolet sterilization reduced losses of eggs to disease and use of glass test tubes as hatching vessels allowed for constant rolling of the eggs and easy viewing and access. The systems occupied little space (< 1 m2) and could accommodate up to 30 treatments of eggs.
The use of egg baskets allowed for rapid and uniform heat shocking of multiple treatments of Nile tilapia eggs for polyploidy induction experiments. Tetraploidy and triploidy were induced by application of 42.8 ± 0.2 C heat shocks to eggs for 3 min at 22, 24, 26 and 28 min AF. Shocks applied at 24 min AF resulted in the highest tetraploid yield (10%) while shocks applied at 22 min AF resulted in the highest triploid yield (7%). This is the first study to report the simultaneous induction of tetraploidy and triploidy in Nile tilapia within treatments from a single spawn and raises questions about technical, maternal and genetic effects on synchrony of zygotic development in this species.
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