Cryopreservation of stallion spermatozoa with different glycerol concentrations

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62157124%3A16170%2F20%3A43878650" target="_blank" >RIV/62157124:16170/20:43878650 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Cryopreservation of stallion spermatozoa with different glycerol concentrations

Popis výsledku v původním jazyce

Glycerol is the most widely used cryoprotectant to freeze stallion spermatozoa. Its effect is not only protective but also toxic and this could be a cause for the variation of stallion sperm motility after thawing. The optimal concentration in extenders is still not clear but almost all conventional extenders use the concentrations between 2% and 6%. The present study aimed to improve the success of cryopreservation of stallion spermatozoa with different glycerol concentrations and to evaluate its effect on stallion sperm motility after thawing. Overall, 228 ejaculates used in this study. Ejaculates from 49 stallions of different breeds were collected, the semen was filtrated to remove gel fraction, macroscopically and microscopically evaluated and only ejaculates with progressive motility higher than 60% (motility I) were used for statistical analysis. After evaluation the ejaculates were centrifuged, the supernatant was removed and spermatozoa were cryopreserved in French diluent (own preparation) supplemented with 2% of centrifuged egg yolk and with different concentrations of glycerol (2.0; 2.5; 4.0 and 6.0%). The choice of concentration of glycerol added to a particular ejaculate was completely random. Based on the concentration used, ejaculates were divided into 4 groups: A (2.0%, 84 ejaculates), B (2.5%, 58 ejaculates), C (4.0%, 69 ejaculates) and D (6.0%, 16 ejaculates). The spermatozoa were packed into 0.5 ml straws and placed in the fridge (4°C) for 2 hours. Then the straws were placed in liquid nitrogen vapor (-80 to -100°C) and after 10 minutes plunged into liquid nitrogen and stored at -196°C for at least 48 hours. The selected straws were individually thawed in a 38°C water bath for 30 seconds before post-freezing analysis. Two progressive sperm motilities were evaluated post-thawing using a phase-contrast microscope (magnification x 400). Because of the large variance of obtained results, nonparametric correlation tests were used for statistical analysis. The Spearmen/Kendall rang correlation test demonstrated a strong relationship between glycerol concentration and motility II but no relationship was found between glycerol concentration and motility III. After the statistical evaluation using the Steel-Dwass test, it was evident that the highest motility II was found in group C and the highest motility III in group D. Because the Spearman and Kendall rank correlation proved a relationship between motility II and glycerol concentration, it can be stated that in this study, the best glycerol concentration for freezing stallion spermatozoa was found to be at a 4%.

Název v anglickém jazyce

Cryopreservation of stallion spermatozoa with different glycerol concentrations

Popis výsledku anglicky

Glycerol is the most widely used cryoprotectant to freeze stallion spermatozoa. Its effect is not only protective but also toxic and this could be a cause for the variation of stallion sperm motility after thawing. The optimal concentration in extenders is still not clear but almost all conventional extenders use the concentrations between 2% and 6%. The present study aimed to improve the success of cryopreservation of stallion spermatozoa with different glycerol concentrations and to evaluate its effect on stallion sperm motility after thawing. Overall, 228 ejaculates used in this study. Ejaculates from 49 stallions of different breeds were collected, the semen was filtrated to remove gel fraction, macroscopically and microscopically evaluated and only ejaculates with progressive motility higher than 60% (motility I) were used for statistical analysis. After evaluation the ejaculates were centrifuged, the supernatant was removed and spermatozoa were cryopreserved in French diluent (own preparation) supplemented with 2% of centrifuged egg yolk and with different concentrations of glycerol (2.0; 2.5; 4.0 and 6.0%). The choice of concentration of glycerol added to a particular ejaculate was completely random. Based on the concentration used, ejaculates were divided into 4 groups: A (2.0%, 84 ejaculates), B (2.5%, 58 ejaculates), C (4.0%, 69 ejaculates) and D (6.0%, 16 ejaculates). The spermatozoa were packed into 0.5 ml straws and placed in the fridge (4°C) for 2 hours. Then the straws were placed in liquid nitrogen vapor (-80 to -100°C) and after 10 minutes plunged into liquid nitrogen and stored at -196°C for at least 48 hours. The selected straws were individually thawed in a 38°C water bath for 30 seconds before post-freezing analysis. Two progressive sperm motilities were evaluated post-thawing using a phase-contrast microscope (magnification x 400). Because of the large variance of obtained results, nonparametric correlation tests were used for statistical analysis. The Spearmen/Kendall rang correlation test demonstrated a strong relationship between glycerol concentration and motility II but no relationship was found between glycerol concentration and motility III. After the statistical evaluation using the Steel-Dwass test, it was evident that the highest motility II was found in group C and the highest motility III in group D. Because the Spearman and Kendall rank correlation proved a relationship between motility II and glycerol concentration, it can be stated that in this study, the best glycerol concentration for freezing stallion spermatozoa was found to be at a 4%.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

40301 - Veterinary science

Projekt

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Návaznosti

S - Specificky vyzkum na vysokych skolach

Rok uplatnění

2020

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů