| 1. |
- Johannisson, Anders, et al.
(författare)
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Colloidal centrifugation with Androcoll-E (TM) prolongs stallion sperm motility, viability and chromatin integrity
- 2009
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Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 116:1-2, s. 119-128
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Forskningsöversikt (refereegranskat)abstract
- The objective was to investigate the changes in stallion sperm quality (sperm motility, viability, membrane integrity and chromatin integrity) occurring during cool storage, and to study the effect of sperm selection by single layer colloidal centrifugation on these parameters of sperm quality. Spermatozoa from 3 stallions (10 ejaculates, 3-4 per stallion) were selected by centrifugation through a single layer of colloid (SLC). The resulting sperm preparations and the control samples (extended but unselected semen samples) were stored at 5 degrees C for 48 h. Assessments of sperm quality, such as sperm motility, viability (SYBR-14/PI staining), membrane stability (Annexin-V/PI staining) and chromatin integrity, were performed on aliquots of the selected sperm preparations and unselected samples on the day of collection (3 h) and after 24 and 48 h of storage. In the SLC-selected sperm samples, sperm motility, sperm viability, proportions of spermatozoa with normal morphology and with intact chromatin were significantly better than in unselected samples (motility: 77 +/- 4% vs. 64 +/- 8% at 3 h; P less than 0.001; viability: 79.5 +/- 9% vs. 64.7 +/- 9%, P less than 0.001: normal morphology 89 +/- 6% vs. 69 9%; chromatin integrity DFI 11.3 +/- 5% vs. 22.1 +/- 10%). Membrane stability, however, was not different in the SLC-selected and unselected samples (74.6 +/- 8% vs. 69.3 +/- 8%). The deterioration seen in sperm quality in the unselected samples was prevented by SLC, so that sperm viability, membrane stability and chromatin integrity were unchanged in the selected samples by 48 h compared to 3 h (Pless than0.001), whereas the unselected samples were significantly worse by 48 h (Pless than0.001). Furthermore, it should be possible to send an aliquot of a normal insemination dose (i.e. unselected spermatozoa) overnight to a reference laboratory for analysis of both plasma membrane and chromatin integrity. In conclusion, centrifugation of stallion spermatozoa through a single layer of colloid is a useful technique for selecting the best spermatozoa from an ejaculate and, moreover, sperm quality is maintained during storage. (C) 2009 Elsevier B.V. All rights reserved.
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| 2. |
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| 3. |
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| 4. |
- Macias Garcia, B., et al.
(författare)
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Centrifugation on a single layer of colloid selects improved quality spermatozoa from frozen-thawed stallion semen
- 2009
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Ingår i: Animal Reproduction Science. - : Elsevier Masson. - 0378-4320 .- 1873-2232. ; 114:1-3, s. 193-202
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Tidskriftsartikel (refereegranskat)abstract
- The present study attempted to select the subpopulation of stallion spermatozoa that best survived a conventional freezing and thawing procedure, using centrifugation of post-thawed semen samples through a single layer of a glycidoxypropyltrimethoxysilane-coated silica colloid with a species-specific formulation (Androcoll-E (TM)). Sperm motility, sperm chromatin structure, membrane integrity and mitorchondrial membrane potential were studied in filtered and non-filtered spermatozoa. Single-layer centrifugation (SLC) using Androcoll-E (TM) significantly improved all the sperm parameters studied, implying SLC may be a simple approach to improve the quality of frozen-thawed (FT) spermatozoa for AI. (c) 2008 Elsevier B.V. All rights reserved.
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| 5. |
- Macias Garcia, B., et al.
(författare)
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Single-Layer Centrifugation Through Colloid Positively Modifies the Sperm Subpopulation Structure of Frozen-Thawed Stallion Spermatozoa
- 2009
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Ingår i: Reproduction in domestic animals. - : Wiley-Blackwell. - 0936-6768 .- 1439-0531. ; 44:3, s. 523-526
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Tidskriftsartikel (refereegranskat)abstract
- The present study attempted to select the subpopulation of stallion spermatozoa that best survived a conventional freezing and thawing procedure, using centrifugation of post-thawed semen samples through a single layer of a glycidoxypropyltrimethoxysilane-coated silica colloid with a species-specific formulation (Androcoll-E (TM)). After freezing and thawing, four sperm subpopulations were identified, listed as FT1 to FT4. While subpopulations FT1 and FT2 were characterized by low sperm velocity, high velocities characterized the ones called FT3 and FT4. The single-layer centrifugation (SLC)-handled sperm sample was enriched in subpopulation FT3, reaching a proportion of 82.6% of the present spermatozoa, in contrast with the non-filtered control post-thawed semen, where this sperm subpopulation only accounted for 16.3% of the total. It is concluded that in the equine industry, the SLC is a practical, easy-to-perform approach to improve the quality of equine frozen-thawed semen samples.
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| 6. |
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| 7. |
- Morrell, Jane, et al.
(författare)
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Colloidal Centrifugation of Stallion Semen: Changes in Sperm Motility, Velocity, and Chromatin Integrity during Storage
- 2009
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Ingår i: Journal of Equine Veterinary Science. - : WB Saunders. - 0737-0806 .- 1542-7412. ; 29:1, s. 24-32
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Tidskriftsartikel (refereegranskat)abstract
- The current study investigated the changes in sperm quality (motility, velocity, and chromatin integrity) occurring during storage at room temperature or 5 degrees C for up to 48 hours in spermatozoa after extension or single-layer centrifugation (SLC) throught Androcoll-E. In unselected samples, all parameters of sperm quality deteriorated significantly during storage (P less than .01), although the deterioration was faster at room temperature (22-30 degrees C) than for cool storage (P less than .01). The SLC-selected spermatozoa had higher motility, velocity, and chromatin integrity than the overall unselected population (motility: selected 85 +/- 10%, unselected 56 +/- 13%; P less than .001; velocity: selected 85.1 +/- 13 mu m/second, unselected 63.5 +/- 15 mu m/second; P less than .001; and DFI selected 12.2 +/- 4.8 mu m/second; unselected 23.6 +/- 7.4 mu m/second; P less than .001). Furthermore, sperm quality did not deteriorate with storage in the SLC-selected samples, either at room temperature (22-30 degrees C for 24 hours) or cooled to 4 degrees C (for at least 48 hours), whereas a significant deterioration in sperm quality was observed in the unselected sperm samples (P less than .01). Thus, room temperature storage of SLC-selected spermatozoa may be an option for insemination doses from stallions whose spermatozoa do not tolerate cooling. In addition, a new sperm analyzer, the Qualisperm, showed good correlation with subjective motility assessment (r = 0.8, P less than .001), was user-friendly, and provided a reasonable volume of data. This instrument may be a useful adjunct to sperm quality assessment at the study.
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| 8. |
- Morrell, Jane, et al.
(författare)
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Comparison of density gradient and single layer centrifugation of stallion spermatozoa: Yield, motility and survival
- 2009
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Ingår i: Equine Veterinary Journal. - : Wiley-Blackwell. - 0425-1644 .- 2042-3306. ; 41:1, s. 53-58
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Tidskriftsartikel (refereegranskat)abstract
- Reasons for performing study: A new, simpler, technique of colloidal centrifugation has recently been developed, designated single layer centrifugation (SLC). This technique requires evaluation by comparison with a density gradient for its ability to select the best quality spermatozoa and its practicality of use on studfarms. Objective: To compare the effect of 2 methods of colloidal centrifugation, density gradient centrifugation and single layer centrifugation, on stallion sperm motility, yield and survival. using freshly collected extended stallion semen. Methods: Aliquots of extended stallion semen from 10 stallions (38 ejaculates) were processed by the 2 methods of colloidal centrifugation. For both uncentrifuged and centrifuged samples, sperm yield was calculated and subjective sperm motility assessed over several days to provide an estimate of sperm survival. Some stored semen samples, held at TV overnight, were also available for testing. Results: For fresh, extended semen, a similar recovery yield of motile spermatozoa was seen for the 2 methods of preparation for single layers and density gradients, respectively. Sperm motility and survival rate Were significantly improved by colloidal centrifugation compared to unprocessed ejaculate, without any significant difference between methods (SLC vs. gradient). However, the yield was reduced by 18-20% when cold-stored semen was used for centrifugation compared to fresh semen, and more variation between ejaculates was observed than for fresh ejaculates. Again, sperm motility and sperm survival were improved in the centrifuged sperm preparations compared to stored, unprocessed ejaculates. Potential relevance: The 2 colloid centrifugation techniques produce equivalent sperm preparations in terms of sperm quality. However, the SIX method would be more practical and convenient for use in the field.
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| 9. |
- Morrell, Jane, et al.
(författare)
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Lipid peroxidation, assessed with BODIPY-C-11, increases after cryopreservation of stallion spermatozoa, is stallion-dependent and is related to apoptotic-like changes
- 2009
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Ingår i: Reproduction. - 1470-1626 .- 1741-7899. ; 138, s. 55-63
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Tidskriftsartikel (refereegranskat)abstract
- Lipid peroxidation (LPO) of stallion spermatozoa was assessed in fresh semen and in samples of the same ejaculates after freezing and thawing. Particular attention was paid to individual differences in the susceptibility to LPO and its possible relationship with freezability. Innate levels of LPO were very low in fresh spermatozoa but increased after thawing, a change that was largely stallion-dependent. The level of LPO in fresh spermatozoa was not correlated with that of the thawed spermatozoa. Negative correlations existed between LPO and intact membranes post-thaw (r= -0.789, P<0.001), and also between LPO and spermatozoa with high mitochondrial membrane potential (Delta psi m) post-thaw (r= -0.689, P<0.001). LPO was also highly and significantly correlated with caspase activity. The correlation between caspase activity in ethidium positive cells and LPO was r=0.772, P<0.001. This LPO is unlikely to represent, per se, a sign of cryopreservation-induced injury, but it is apparently capable of triggering 'apoptotic-like changes' that could result in the sub-lethal cryodamage often seen among surviving spermatozoa. Reproduction (2009) 138 55-63
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| 10. |
- Morrell, Jane M., et al.
(författare)
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Selection of Boar Spermatozoa Using Centrifugation on a Glycidoxypropyltrimethoxysilane-Coated Silica Colloid
- 2009
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Ingår i: Journal of reproduction and development. - : Society for Reproduction and Development. - 0916-8818 .- 1348-4400. ; 55:5, s. 547-552
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Tidskriftsartikel (refereegranskat)abstract
- Use of sperm separation methods such as density gradient centrifugation for selecting the best spermatozoa for animal breeding is constrained by the problem of dealing with the large volumes of ejaculate produced by the males of some species, such as boars. The purpose Of this Study was to compare density gradient centrifugation (DGC) with centrifugation on a single layer of colloid (SLC) for the preparation of ejaculated boar spermatozoa using Androcoll (TM)-P. There was no difference between the two techniques in terms of sperm motility or duration of motility after selection, and sperm motility was retained for at least 24 h longer in the centrifuged sperm preparations than in controls (uncentrifuged aliquots). Sperm motility was significantly better (Pless than0.001) in the centrifuged sperm preparations (means +/- sd: SLC 79.6 +/- 8.1% and DGC 74.2 +/- 12.0%) than in the uncentrifuged controls (62.9 +/- 12.7%). The mean yield of motile spermatozoa for SLC was 67.5 +/- 25.6%, and for DGC it was 59.6% +/- 22.3%, (not significant, ns). Sperm survival was significantly increased by colloid centrifugation (control 3.1 +/- 0.3 days, SLC 5.5 +/- 0.79 days, DGC 5.75 +/- 0.62 days; Pless than0.001 for uncentrifuged versus centrifuged, SLC vs. DGC, ns). Moreover, boar spermatozoa could be stored for 24 h before centrifugation without: any detrimental effect on sperm motility or duration of motility. In a further experiment, larger volumes of ejaculate were processed easily on a modified SLC, indicating that this method may be practical for processing large volumes of boar ejaculates.
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