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. 1996 Dec 1;37(4):461–470. doi: 10.1186/BF03548086

Influence of Sperm Number Per Straw on the Post-Thaw Sperm Viability and Fertility of Swedish Red and White A.I. Bulls

Påverkan av spermιeantalet per strån på uptιnιngs vιabιhtet och fertιlιtet hos SRB-semιn tjurar

A Januskauskas 110, L Söderquist 110, M G Håård 210, M Ch 210, N Lundeheim 310, H Rodriguez-Martinez 110,✉
PMCID: PMC8064005  PMID: 9050279

Abstract

Semen from 5 Swedish Red and White bulls, approved in the fertility and progeny testing programme of a bull center, was split-frozen to produce straws with 15 or 10×106 spermatoza each (control and treatment dose, respectively). Post-thaw sperm viability was evaluated by visual assessment of sperm motility (MOT), measurement of ATP (Adeinosin Tri Phosphate) contents by luminometry, assessment of membrane integrity with combined fluorophore probes [Calcein AM (CAM)/Ethidium homodimer (EthD-1)] and by using a hyposmotic swelling test (ORT). The straws were used for a total of 16651 artificial inseminations (A.I.). No statistically significant difference was recorded between the 2 treatments for any of the post-thaw sperm viability parameters. In addition, a significant bull effect was evident for most post-thaw sperm traits assayed. Significant variation in overall fertility (56-days NRR) was recorded among the bulls used. A.I. with a reduced number of spermatozoa (10xl06/straw) resulted in a 2%-units decrease (n.s.) compared with controls (67.8% ± 4.8%, means ± SD) in overall fertility. In the control split-sample (15×l06 spermatozoa/straw), MOT did not show any statistically significant correlation with fertility (r = 0.41, p = 0.07). However, MOT was correlated with the percentage of spermatozoa depicting progressive motility (category A1, r = 0.45, p<0.05) as assessed with CAM/EthD-1. The latter was correlated with ATP contents (r = 0.57, p0.01), expressed as the percentage of viable spermatozoa. Both CAM/EthD-1 and ATP contents showed a statistically significant correlation with ORT (r = 0.45, p<0.05 and r = 0.61, p<0.05, respectively). In the straws with the reduced sperm number (10×l06 spermatozoa), post-thaw motility was significantly correlated with fertility (r = 0.50, p<0.05) and ATP-total contents (r = 0.48, p<0.05). CAM/EthD-1 and ATP contents (as million viable spermatozoa) were significantly correlated (r = 0.47, p<0.05). CAM/EthD-1 was significantly correlated with ORT (r = 0.48, p<0.05). In conclusion, the results indicated that the freezing-thawing of straws with a reduced sperm concentration (110×106 spermatozoa) did not alter the post-thaw viability or overall fertility of the bull semen used. However, in view of the significant bull effect found in the limited population studied, we recommend that such a reduction in sperm number/straw be based on the fertility of the bull in question.

Keywords: motility, ATP, membrane integrity, hypo-osmosis

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Acknowledgments

The technical assistance provided by Mrs. A. Rikberg and Mrs. K. Selin-Wretling is gratefully acknowledged. The authors are also very grateful to the personnel at the semen processing laboratory, Svensk Avel, Skara. Dr. Januskauskas holds a post-graduate fellowship from the Royal Swedish Academy of Forestry and Agriculture and the Swedish University of Agricultural Sciences. This work received financial support from Stiftelsen Lantbruksforskning, Sweden.

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