Delivery of 17 -Methyltestosterone through Tilapia Embryos by Electrical Pulse Inductions for Commercial Sex Reversal
Abstract
The present study proposes an alternative technique of electrical sex reversal to enhance all-male sex reversal (MSR) of Nile tilapia using transient square pulse-electric fields for commercial benefits focusing on the study of mechanisms of electro-permibilization of 17 -Methyltestosterone (MT) androgen hormones through induced membrane of tilapia’s embryo (Oreochromis niloticus L., Chitralada), during segmentation-pharyngula (2-3 days post fertilization). The technique were carried out for large-scale inductions of embryos suspensions with MT concentrations of 1,000 – 100,000 µg l-1using square-pulse electric fields of amplitudes 100-500 volts (electric field intensities of 25-125 kV m-1), 100 µs pulse-period (50:50 mark space ratio) and 3 square pulses, respectively. It was found that the most suitable conditions of 350 V with MT concentration of 5,000 µg l-1 gave the maximum rate of male-sex reversal rate (MSR) of 89.42%±2.28%, hatching rate (HR) of 93.33%±0.58% and survival rate (SR) of 87.49%±1.71%, respectively. The off-spring rate (OSR) of male and female in the control group were 59.71%±3.00% and 46.33%%±1.53%, respectively, with hatching rate (HR) of 91.33%±1.53% and survival rate (SR) of 85.08% ± 4.16% (increasing MSR of 29.71%). Verifications of MT delivery were carried out through LC-MS/MS technique and RNA expression profiles by extraction of total RNA from induced MT embryos. Genes of male-sex development (up-regulated genes) of amh, ar, dmrt1, bmp15, gfd9 and down-regulated genes of gsdf (where house keeping genes of rpl13a) were investigated by Quantitative real-time PCR. The induced MT embryos with transient square pulse-electric fields showed expression of male-sex development genes. Keywords : tilapia; sex reversal rate; pulse electric fields; 17 -Methyltestosterone; geneReferences
Bunthawin, S., Wanichapichart, P. and Gimsa, J. (2007). An investigation of dielectric properties of biological cells
using RC-model, Songklanakarin Journal of Science and Technology, 29(4), 1163-1181.
Bunthawin, S., Wanichapichart, P., Tuantranont, A. and Coster, H. G. L. (2010). Dielectrophoretic spectra of
translational velocity and critical frequency for a spheroid in traveling electric fields, Biomicrofluidics, 4(014102), 1-13 (Doi:10.1063/1.3294082).
Bunthawin, S. Raymond J. R. and Wanichapichart, P. (2011). Dielectrophoresis of Tetraselmis sp., a unicellular
green alga, in travelling electric fields analyzed using the RC model for a spheroid. Songklanakarin Journal
of Science and Technology, 33 (5), 585-597.
Bunthawin, S. and Raymond J. R. (2013). Simulation of translational dielectrophoretic velocity spectra of
erythrocytes in traveling electric field using various volume models, Journal of Applied Physics, 113(014701), 1-10 (Doi:10.1063/1.4773326).
Bunthawin, S., Sornsilpa, T., Tuantranont, A., Jaruwongrangsri, K. and Raymond J.R. (2015). Monosex-male
sex reversal of Nile tilapia embryos using pulse-electric field inductions. Computational and Theoretical Nanoscience, 12, 1-5.
Chang, D. C., Chassy, B. M., Saunders, J. A. and Sowers, A. E. (1992). Guide to electroporation and
electrofusion. California: Academic Press.
Chervinski, J. and Rothbard, S. (1982). An aid in manually sexing tilapia. Aquaculture, 26, 389.
Department of Fisheries. (2017). Fisheries Statistics of Thailand. In Annual Report No.9/2019. (pp. 40-74).
Bangkok: Ministry of Agriculture and Cooperatives.
McAndrew, B. J. (1993). Sex control in Tilapiines. London: Blackwell.
Pohl, H. A. (1978). Dielectrophoresis: The behavior of neutral matter in non-uniform electric fields.
Cambridge: Cambridge University Press.
Srisakultiew, P. (2003). Study on status of Nile tilapia sex reversal to reduce cost. In Complete research
report. (pp. 11-12). Bangkok: Thailand Research Fund.
Tsong, T. Y. (1991). Electroporation of cell membranes. Biophysical Journal, 60, 297-306.
using RC-model, Songklanakarin Journal of Science and Technology, 29(4), 1163-1181.
Bunthawin, S., Wanichapichart, P., Tuantranont, A. and Coster, H. G. L. (2010). Dielectrophoretic spectra of
translational velocity and critical frequency for a spheroid in traveling electric fields, Biomicrofluidics, 4(014102), 1-13 (Doi:10.1063/1.3294082).
Bunthawin, S. Raymond J. R. and Wanichapichart, P. (2011). Dielectrophoresis of Tetraselmis sp., a unicellular
green alga, in travelling electric fields analyzed using the RC model for a spheroid. Songklanakarin Journal
of Science and Technology, 33 (5), 585-597.
Bunthawin, S. and Raymond J. R. (2013). Simulation of translational dielectrophoretic velocity spectra of
erythrocytes in traveling electric field using various volume models, Journal of Applied Physics, 113(014701), 1-10 (Doi:10.1063/1.4773326).
Bunthawin, S., Sornsilpa, T., Tuantranont, A., Jaruwongrangsri, K. and Raymond J.R. (2015). Monosex-male
sex reversal of Nile tilapia embryos using pulse-electric field inductions. Computational and Theoretical Nanoscience, 12, 1-5.
Chang, D. C., Chassy, B. M., Saunders, J. A. and Sowers, A. E. (1992). Guide to electroporation and
electrofusion. California: Academic Press.
Chervinski, J. and Rothbard, S. (1982). An aid in manually sexing tilapia. Aquaculture, 26, 389.
Department of Fisheries. (2017). Fisheries Statistics of Thailand. In Annual Report No.9/2019. (pp. 40-74).
Bangkok: Ministry of Agriculture and Cooperatives.
McAndrew, B. J. (1993). Sex control in Tilapiines. London: Blackwell.
Pohl, H. A. (1978). Dielectrophoresis: The behavior of neutral matter in non-uniform electric fields.
Cambridge: Cambridge University Press.
Srisakultiew, P. (2003). Study on status of Nile tilapia sex reversal to reduce cost. In Complete research
report. (pp. 11-12). Bangkok: Thailand Research Fund.
Tsong, T. Y. (1991). Electroporation of cell membranes. Biophysical Journal, 60, 297-306.
Downloads
Published
2021-05-05
Issue
Section
Research Article
