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Motility and fertilizing capacity of fresh and frozen-thawed spermatozoa in sturgeons Acipenser Baeri and A. ruthenus
Authors:L I Tsvetkova  J Cosson  O Linhart  R Billard
Institution:Laboratory of Cryobiology, All-Russian Institute of Pond Fisheries, Rybnoye, Dmitrov Region, Moscow Area, Russia;CNRS URA 671, Station Marine, Villefranche sur Mer, France;Research Institute of Culture and Hydrobiology, University of South Bohemia, Department of Fish Genetics and Breeding, Vodnany, Czech Republic;Laboratoire d'Ichtyologie, Museum National d'Histoire Naturelle, Paris Cedex, France
Abstract:The Siberian sturgeon ( Acipenser baeri ) and the sterlet ( A. ruthenus ) were injected with dried sturgeon pituitary (2 mg kg−1), yielding 24 h later respectively 1.71 ± 0.5 and 1.65 ± 0.5 1011 spermatozoa kg−1 body weight. Spermatozoa were best activated with a solution of Tris HC1 50 mM, pH 8.0. The percentage of activated cells was 88 ± 4.4 in A. baeri (n = 5) and 68 ± 19 in A. ruthenus. (n = 5). In A. baeri , immediately after activation, the beat frequency of the flagellum and the mean velocity were in the range of 48–52 Hz and 100–300 μm−1s, respectively. The beat frequency declined to 15 Hz at 30–40 and velocity to 100 μm s−1 at 60 s post-activation. Only a small percentage of the spermatozoa remained motile after 3–4 min. In all cases spermatozoa showed mostly quasi-linear trajectories. Sperm was frozen in liquid nitrogen vapor immediately after dilution 1 v: 1 v in a cryopreservation medium (23.4 mM saccharose, 118 mM Tris-HCl pH 8.0, 20% egg yolk to which 15% DMSO were added). After fast-thawing procedure (25 s at 40°C), the percentage of motile spermatozoa (once activated in 118 mM Tris-HCl, pH 8.0) decreased to 23 ± 8.8 in A. baeri and to 15 ± 11 in A. ruthenus. The fertilizing capacity also decreased: 53 ± 8.3% vs. 89 ± 7.6% in control (P < 0.05) in A. baeri and 23 ± 11% vs 53 ± 8.3% in control (P < 0.05) in A. ruthenus. The motility pattern of the surviving frozen/thawed spermatozoa was the same as in fresh spermatozoa.
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