Effect of dialysis or centrifugation on post-thaw motility and fertility of Santa Gertrudis bull semen collected by electroejaculation

Tibary, Ahmed; Graham, E.F.; Asri, Alfian; Boukhliq, R.; Deyo, R.

doi:10.1016/0093-691x(90)90550-d

Cited by 6 publications

(4 citation statements)

References 6 publications

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“…Furthermore, in bulls reared in the field, semen is collected by electroejaculation. The ejaculates obtained by this method differ in terms of the quantity and quality of SP from those obtained using artificial vagina, a fact that considerably alters the SP/sperm ratio in semen [8,9].…”

Section: Introductionmentioning

confidence: 99%

Effect of seminal plasma removal before cryopreservation of bovine semen obtained by electroejaculation on semen quality and in vitro fertility

et al. 2017

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Cryopreservation of bull semen is a common biotechnology procedure in cattle breeding. However, when the ejaculate is obtained by electroejaculation, wide variation is observed in the sperm/seminal plasma (SP) ratio that can affect the freezability of semen in this species. The removal of SP may improve the quality of frozen bull semen. The objective of this study was to evaluate the effect of SP removal from the ejaculate on the cryopreservation of semen from 38 Nellore bulls collected by electroejaculation. After collection, the ejaculate was divided into three aliquots: (1) control (N) diluted to a concentration of 60 × 10 spermatozoa/mL and frozen with SP; (2) centrifugation (C) at ×600g for 10 minutes and the pellet resuspended and frozen at the same concentration as N; and (3) filtration (F) through SpermFilter and sperm recovered and frozen at the same concentration as N. After thawing, sperm kinetics, plasma and acrosome membrane integrity, mitochondrial membrane potential, oxidative stress, and in vitro fertility were evaluated. Statistical analysis was performed using the SAS 9.2 package, and differences were considered significant when P < 0.05. Higher average path velocity and straight-line velocity were observed in the groups submitted to SP removal compared to the control group (P < 0.01). In contrast, filtered samples exhibited higher beat cross frequency, straightness, and linearity compared to the other groups. Plasma membrane integrity was reduced when SP was removed, but lower oxidative stress was observed in groups C and F (34.91 ± 2.95% and 31.63 ± 2.95%, respectively) compared to group N (57.39 ± 2.95%). However, the percentage of hatched blastocysts was similar in the N and F groups (21.22 ± 1.05% and 24.00 ± 1.05%, respectively) and higher compared to group C (18.83 ± 1.05%). In conclusion, removal of SP by centrifugation for bull semen freezing reduced the rate of in vitro-produced embryos, whereas filtration of prefrozen semen was found to be an efficient alternative in terms of semen freezability and in vitro production of bovine embryos.

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Section: Introductionmentioning

confidence: 99%

Effect of seminal plasma removal before cryopreservation of bovine semen obtained by electroejaculation on semen quality and in vitro fertility

et al. 2017

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“…Not only did sperm sort more efficiently when seminal plasma was removed, but post-thaw motility also increased by 17%. An increase in post-thaw motility was also seen by Tibary et al (1990) after sperm cryopreservation with seminal plasma removed.…”

Section: Discussionmentioning

confidence: 77%

Removing Seminal Plasma Improves Sex-Sorting of Bovine Sperm.

Burroughs

Graham

Lenz

et al. 2011

Biology of Reproduction

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REMOVING SEMINAL PLASMA IMPROVES SEX-SORTING OF BOVINE SPERMExperiments for this thesis evaluated how characteristics of bovine ejaculates affect efficiency of sorting X-from Y-chromosome bearing sperm by flow cytometry/cell sorting, and further, document that removal of seminal plasma improves efficacy of sexsorting bovine sperm. In Experiment I, ejaculates were collected by artificial vagina, two each from 10 bulls with an average of one hour between collections. Semen was centrifuged to separate sperm from seminal plasma, and then sperm were re-diluted to 160x10 6 sperm per ml with staining TALP (Tyrode's albumin, lactate, pyruvate) and 0, 5, 10 or 20% seminal plasma from the same ejaculate or reciprocally from first/second ejaculates. Following incubation with Hoechst 33342, sperm were sorted and analyzed with a flow cytometer/cell sorter. The % live-oriented sperm was higher for treatments with 0% seminal plasma (64.4%) than 5 (59.6%), 10 (59.0%) and 20% (57.8%) seminal plasma (p<0.01). The % live-oriented sperm was higher for second (63.0%) than first ejaculates (56.2%). Sort rate was higher for samples with 0% seminal plasma (p<0.05).The percentages of membrane-impaired sperm were lower for 0% (16.5%) than 5 (21.9%), 10 (23.6%) or 20% (23.4%) seminal plasma (p<0.003), and for second iii ejaculates (18.2%) compared to first ejaculates (25.9%). Whether seminal plasma was from first versus second ejaculates had no effect (p>0.1). Effects of seminal plasma originating from bulls different from those whose sperm were sorted was evaluated in Experiment II. Semen collection and initial analysis were performed as in Experiment I and for all successive experiments. Seminal plasma from ejaculates of 6 bulls (3 that sorted well; 3 that sorted poorly) used in Experiment I was used with sperm collected for Experiment II as well as seminal plasma from sperm donors. Sperm were stained with Hoechst 33342 as in Experiment I, but with 10% seminal plasma in staining TALP.Only seminal plasma from one bull resulted in differences for any sorting parameters; 31% of sperm were membrane-impaired with seminal plasma from this bull compared to 16-19% for seminal plasma from other bulls (p<0.05). Sort rate was decreased (3.02x10 3 sperm per sec) compared to a range of 3.64-3.97x10 3 sperm per sec with the other seminal plasmas (p<0.05). This may have been due to contamination of the original semen sample, since this effect was only seen with one of six bulls.Experiment III investigated effects of adding bovine serum albumin (BSA) during staining with Hoechst 33342 using semen from 10 bulls. BSA was added at 0, 0.3 or 0.9% to staining TALP with either 0 or 10% seminal plasma. Although BSA has been shown to be beneficial to sperm, there was a possibility that BSA would bind Hoechst 33342, thereby reducing sorting efficiency. There was no evidence of either effect; however, sperm with 0% seminal plasma had higher % live oriented cells (65%) than sperm incubated with 10% seminal plasma (61%; p<0.05). In addition, samples conta...

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“…Similarly, Tibary et al . () reported increased post‐thaw sperm motility in samples where the seminal plasma had been removed. Therefore, seminal plasma impairs the ability of bull sperm to be sex‐sorted and cryopreserved.…”

Section: Discussionmentioning

confidence: 99%

Removing seminal plasma improves bovine sperm sex‐sorting

et al. 2016

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Bull ejaculates with sperm concentrations of less than 1 billion sperm sort poorly for sex chromosomes, but whether this is because of the sperm concentration or the concomitant seminal plasma content has not been elucidated. Experiments were conducted to determine why ejaculates with lower sperm concentrations sort poorly and develop a protocol to increase sorting efficiency. In Experiment I, spermatozoa at 160 or 240 × 10 sperm/mL were stained at 49, 65 or 81 μm Hoechst 33342 with 0 or 10% seminal plasma and then sex-sorted. In Experiment II, seminal plasma was adjusted to create samples with sperm concentrations of 0.7, 1.4 and 2.1 × 10 sperm/mL, prior to sex-sorting. In Experiment III, spermatozoa were diluted to 0.7, 1.4 and 2.1 × 10 sperm/mL using TALP containing 0 or 10% seminal plasma prior to sex-sorting and cryopreservation. In Experiment I, the optimal staining combination was 160 × 10 sperm/mL stained with 65 μm Hoechst 33342 and no seminal plasma. In Experiment II, the percentages of membrane-impaired sperm were lower for sample concentrations of 2.1 × 10 sperm/mL (15%) than for samples at 1.4 × 10 (17%) or 0.7 × 10 sperm/mL (18%; p < 0.01). The X sort rate was slower for samples stored at 0.7 × 10 sperm/mL (3.45 × 10 sperm/sec) than for samples stored at 1.4 × 10 and 2.1 × 10 sperm/mL (3.85 and 3.94 × 10 sperm/sec, respectively; p < 0.05). In Experiment III, samples containing 0% seminal plasma had higher percentages of live-oriented cells (54 vs. 50%; p < 0.05), fewer dead sperm (19 vs. 22%; p < 0.01) and higher post-thaw motility (41 vs. 35%; p < 0.05) than samples containing 10% seminal plasma. Ejaculates with high sperm concentrations result in superior sorting because these samples have less seminal plasma during staining than ejaculates with lower initial sperm concentrations as all samples are diluted to 160 × 10 sperm/mL for staining. Therefore, sorting efficiency appears to be affected by seminal plasma concentration, not by the original sperm concentration.

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Effect of dialysis or centrifugation on post-thaw motility and fertility of Santa Gertrudis bull semen collected by electroejaculation

Cited by 6 publications

References 6 publications

Effect of seminal plasma removal before cryopreservation of bovine semen obtained by electroejaculation on semen quality and in vitro fertility

Effect of seminal plasma removal before cryopreservation of bovine semen obtained by electroejaculation on semen quality and in vitro fertility

Removing Seminal Plasma Improves Sex-Sorting of Bovine Sperm.

Removing seminal plasma improves bovine sperm sex‐sorting

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