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The effect of Equex STM in freezing media on post thaw motility, viability and dna integrity of frozen - thawed ram spermatozoa


Sterbenc, N; Kosec, M; Bollwein, H; Klinc, P (2014). The effect of Equex STM in freezing media on post thaw motility, viability and dna integrity of frozen - thawed ram spermatozoa. Slovenian Veterinary Research, 51(1):35-42.

Abstract

In this study we investigated the effect of Equex STM® on quality and in-vitro survival of ram spermatozoa frozen in Tris egg yolk based extender. Ejaculates from 6 crossbreed rams were frozen according to the standard procedure after two step dilution with Tris-egg yolk extender (1). The second extender, added to the semen at 5°C, contained 14% of glycerol and was supplemented with detergent 0.75% Equex STM® (group OEP) or contained no detergent (control group). After thawing the samples were incubated in a water bath at 37°C and analysis were performed 10 minutes, 6, 12 and 24 hours later. Motility and the viability (Viadent®) of the semen were analysed with Hamilton Thorne Biosciences, Version 12.3 and membrane integrity with HOS (hypoosmotic swelling test). DNA fragmentation (DFI %) of F/T spermatozoa was analyzed 10 minutes and 3 hours after thawing using sperm chromatin structure assay (SCSATM). The sperm membrane integrity was analysed 15 minutes and 3 hours after thawing by Sybr-14/PI test. Percentage of motile spermatozoa was significantly higher in OEP group in comparison to control group at 0, 6, 12 and 24 h (P<0.001). Viability of spermatozoa was significantly higher (P<0.001) in OEP compared to control group in all analysed times after thawing. Percentage of HOS positive spermatozoa was significantly higher in OEP compared to control group respectively for 0 (P=0.001), 6 (P=<0.001), 12 and 24 h (P=0.002) after thawing.

Abstract

In this study we investigated the effect of Equex STM® on quality and in-vitro survival of ram spermatozoa frozen in Tris egg yolk based extender. Ejaculates from 6 crossbreed rams were frozen according to the standard procedure after two step dilution with Tris-egg yolk extender (1). The second extender, added to the semen at 5°C, contained 14% of glycerol and was supplemented with detergent 0.75% Equex STM® (group OEP) or contained no detergent (control group). After thawing the samples were incubated in a water bath at 37°C and analysis were performed 10 minutes, 6, 12 and 24 hours later. Motility and the viability (Viadent®) of the semen were analysed with Hamilton Thorne Biosciences, Version 12.3 and membrane integrity with HOS (hypoosmotic swelling test). DNA fragmentation (DFI %) of F/T spermatozoa was analyzed 10 minutes and 3 hours after thawing using sperm chromatin structure assay (SCSATM). The sperm membrane integrity was analysed 15 minutes and 3 hours after thawing by Sybr-14/PI test. Percentage of motile spermatozoa was significantly higher in OEP group in comparison to control group at 0, 6, 12 and 24 h (P<0.001). Viability of spermatozoa was significantly higher (P<0.001) in OEP compared to control group in all analysed times after thawing. Percentage of HOS positive spermatozoa was significantly higher in OEP compared to control group respectively for 0 (P=0.001), 6 (P=<0.001), 12 and 24 h (P=0.002) after thawing.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Veterinary Clinic > Department of Farm Animals
Dewey Decimal Classification:570 Life sciences; biology
630 Agriculture
Date:2014
Deposited On:12 Feb 2015 10:06
Last Modified:08 Dec 2017 11:20
Publisher:University of Ljubljana
ISSN:1580-4003
Official URL:http://www.slovetres.si/
Related URLs:http://connection.ebscohost.com/c/articles/95805335/effect-equex-stm-freezing-media-post-thaw-motility-viability-dna-integrity-frozen-thawed-ram-spermatozoa
http://www.cabdirect.org/abstracts/20143171329.html

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