Abstract
In this study, we trialed 6-dimethylaminopurine (6-DMAP) chemical shocks to induce meiosis I or meiosis II Pacific White shrimp, Litopenaeus vannamei, triploids for the first time, and cold temperature shocks to induce meiosis II L. vannamei triploids as done previously. Inductions were performed on 37 spawnings in total with experiments being progressively designed in a factorial manner to allow optimization of induction parameters. Treatment with a 200-μm 6-DMAP final concentration at 1 min post-spawning detection for a 6 to 8 min duration resulted in the most consistent induction of chemically induced meiosis I triploids while treatment at 7 min 30 s post-spawning detection for a 10-min duration resulted in the most consistent induction of chemically induced meiosis II triploids. A cold temperature shock of 11.7°C to 13.25°C (final treatment temperature; spawning water temperature 28.5°C) applied at 8 min post-spawning detection for a 4 to 10 min duration resulted in the most consistent induction of cold-temperature-induced meiosis II triploids. 6-DMAP shocks resulted in meiosis I induction rates from 29% to 100% in unhatched embryos and 50% in nauplii, and meiosis II induction rates from 65% to 100% in unhatched embryos and 52% to 100% in nauplii. Cold shocks resulted in induction rates from 5% to 100% in unhatched embryos and nauplii. Confocal microscopy analysis of embryos revealed that there are major developmental abnormalities in a large proportion of later stage triploid L. vannamei embryos compared to their diploid sibling controls. Despite this, however, some triploid embryos did appear normal and both shock agents induced small numbers of viable triploid L. vannamei nauplii which were successfully reared to protozoeal stage 3 as confirmed by flow cytometry. Triploids beyond this life-history stage were not observed in the present study as confirmed by flow cytometry at mysis stages. This study adds to our knowledge base of triploid induction in L. vannamei and further highlights the inherent difficulties with triploid embryonic and larval viability in this species.
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Acknowledgments
The authors would like to thank Bob Cartell, Starr Kapau, and Scott Naguwa from The Oceanic Institute for their assistance with spawning system set-up and animal handling, and Karen Selph at the University of Hawaii for her contributions to ploidy analysis. This project was funded through the Australian Academy of Sciences International Science Linkages program, Scientific Visits to North America and the CSIRO Food Futures National Research Flagship.
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Sellars, M.J., Arce, S.M. & Hertzler, P.L. Triploidy Induction in the Pacific White Shrimp Litopenaeus vannamei: An Assessment of Induction Agents and Parameters, Embryo Viability, and Early Larval Survival. Mar Biotechnol 14, 740–751 (2012). https://doi.org/10.1007/s10126-012-9439-2
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DOI: https://doi.org/10.1007/s10126-012-9439-2