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Improvement of common carp artificial reproduction using enzyme for elimination of egg stickiness

Published online by Cambridge University Press:  15 October 2003

Otomar Linhart
Affiliation:
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
Marek Rodina
Affiliation:
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
David Gela
Affiliation:
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
Martin Kocour
Affiliation:
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
Martha Rodriguez
Affiliation:
Autonomus University Metropolitane, Mexico City 04960, Mexico
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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 23672 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 withα-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.

Type
Research Article
Copyright
© Elsevier, IRD, Inra, Ifremer, Cemagref, 2003

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