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Comparison of two different centrifugation extenders for preservation of frozen equine semen
Author(s) -
Leão Freitas M.,
Silva Bouéres C.,
Gonçalves de Oliveira F. J.,
Oliveira Viu M. A.,
Arruda de Oliveira R.
Publication year - 2015
Publication title -
equine veterinary education
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.304
H-Index - 31
eISSN - 2042-3292
pISSN - 0957-7734
DOI - 10.1111/eve.12357
Subject(s) - extender , semen , skimmed milk , acrosome , sperm , centrifugation , semen extender , cryopreservation , sperm motility , chromatography , andrology , membrane integrity , chemistry , food science , medicine , biology , membrane , biochemistry , embryo , organic chemistry , polyurethane , microbiology and biotechnology
Summary This study compares a commercial semen extender (control group) to ultra high temperature ( UHT ) skimmed milk (treatment group) used during centrifugation for subsequent cryopreservation of equine semen. Following post‐thawing of semen samples parameters measured included motility, sperm motion kinetics (using computerised assisted semen analysis) as well as acrosome and plasmatic membrane integrity (using fluorescent dyes). After collection and analysis, the sperm‐rich fraction was divided and diluted with either: control (1:1 dilution in a skimmed milk‐glucose extender) or treatment (1:1 dilution in UHT skimmed milk). The milk used in this experiment was of the same source, commercial brand, of only one lot. After dilution, samples were subjected to centrifugation at 600 g for 10 min and sperm pellets were resuspended in a freezing extender to a concentration of 200 × 10 6 cells/ml. Aliquots were packed into 0.5 ml straws placed in a stainless steel support and kept inside the refrigerator (5°C) for 20 min. Subsequently, these straws were placed at a height of 6 cm over liquid nitrogen for 20 min in an isotherm box. No significant differences were observed in total sperm motility (42.71 vs. 38.29%), progressive sperm motility (12.29 vs. 7.86%), plasma membrane integrity (53.43 vs. 60.14%) or acrosomal membrane integrity (93.29 vs. 93.71%) with a P>0.05 calculated between the control and the treatment groups, respectively. Considering that UHT skimmed milk has a lower cost than the commercial semen extender, this could be an option used during the centrifugation protocol to decrease the expense of the equine semen cryopreservation process and increase shelf life.