Improvement of common carp artificial reproduction using enzyme for elimination of egg stickiness |
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| Institution: | 1. Joint Laboratory of Genetics, Physiology and Reproduction of Fish, Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, University of South Bohemia, Research Institute of Fish Culture and Hydrobiology, 38925 Vodnany, Czech Republic;2. Autonomus University Metropolitane, Mexico City 04960, Mexico;1. Department of Hepatobiliary and Pancreatic Surgery, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China;2. Collaborative Innovation Center for Cancer Medicine, Zhejiang University, Guangzhou, China;1. Instituto de Química Aplicada. Universidad del Papaloapan. Circuito Central 200, colonia Parque Industrial, Tuxtepec, Oaxaca, México 68301;2. Programa de Posgraduados en Ingeniería de la Producción y Sistemas (PPGEPS). Pontificia Universidad Católica de Paraná (PUCPR). Inmaculada Concepción 1155(80215-901). Curitiba (PR), Brasil;3. Instituto Universitario de Automática e Informática Industrial. Universidad Politécnica de Valencia. Camino de Vera s/n 46022. Valencia, España;4. Departamento de Ingeniería Química y Bioquímica. Instituto Tecnológico de Veracruz. Av. Miguel Ángel de Quevedo 2779. Veracruz, Ver., México 91860;1. Department of Neurosurgery, Antwerp University Hospital, Edegem, Belgium;2. Faculty of Medical and Health Sciences, University of Antwerp, Wilrijk, Belgium;3. Department of Neurology, Antwerp University Hospital, Edegem, Belgium;4. Department of Radiology, Antwerp University Hospital, Edegem, Belgium;5. Department of Neurosurgery, St. Augustinus Hospital, Wilrijk, Belgium;1. School of Mineral Resources Engineering, Applied Mathematics and Computers Laboratory, Technical University of Crete, University Campus, 73132 Chania, Crete, Greece;2. Institute of Applied and Computational Mathematics, Foundation for Research and Technology - Hellas, 70013 Heraklion, Greece;3. Department of Aerospace Engineering, Daytona Beach College of Engineering, Embry-Riddle Aeronautical University, 600 S. Clyde Morris Blvd. Daytona Beach, FL 32114, USA |
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| Abstract: | This study summarizes optimization of techniques for common carp artificial propagation including improvements of activation solution (AS), the process of insemination, and elimination of egg stickiness. The optimum gamete ratio for good fertilization and hatching rate ranged from 8490 to 23 672 spermatozoa per egg, when dechlorinated tap water was used. Optimal ratio between eggs (weight in g) and AS (in ml) was defined as 1:1 to 1:2. Different concentrations of AS such as NaCl from 0 to 34 mM (0–68 mOsmol kg–1) did not change fertilization and hatching rates. An AS adopted for carp spermatozoa (45 mM NaCl, 5 mM KCl, 30 mM Tris–HCl, pH 8) was compared with other saline AS; only the 51 mM (102 mOsmol kg–1) NaCl solution decreased fertilization and hatching rate. The AS containing 20 mM Tris–HCl at pH 9 increased fertilization and hatching rates compared to dechlorinated tap water of pH 7 or to AS of pH 6 and 7. Adhesiveness from the eggs was successfully removed by incubation in Alcalase DX (PLN 04715) using two successive steps with different enzyme concentrations. The first step with an enzyme concentration of 2 ml l–1 was applied from 8 to 20 min after fertilization. Later in a second step, the best time for application of alcalase enzyme at a concentration of 20 ml l–1 was for 45 and 60 s at 20 min after fertilization leading to fertilization and hatching rates of 80–87%. The α-Chymotrypsin (EC 3.4.21.1. Merck) was also found effective for elimination of stickiness. Results with α-Chymotrypsin enzyme indicate that the response to success in elimination of stickiness is highly variable mainly due to differences in the environment, quality of water and carp strains. |
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