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- Abraham, Maria Celina, et al.
(författare)
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Effect of sperm preparation on development of bovine blastocyst in vitro
- 2016
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Ingår i: Zygote. - 0967-1994 .- 1469-8730. ; 24, s. 825-830
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Tidskriftsartikel (refereegranskat)abstract
- colloids has been used to select normal sperm for assisted reproduction in several species. Animal models can sometimes be used as a preliminary step to investigate sperm preparation methods that are potentially of use for human fertility treatments. In this study bovine semen was prepared using three variants of the single-layer centrifugation sperm selection technique (Small, Mini, Mini-EP) with Bovicoll (Androcoll-B). Computer-assisted sperm motility analysis, the hypo-osmotic swelling test, and the sperm chromatin structure assay were performed on unselected (control) and SLC-selected sperm samples. Mini and Mini-EP gave the highest yield of motile spermatozoa, progressive motility and membrane integrity. In vitro fertilization trials were performed to investigate the fertilizing ability of the frozen-thawed bovine spermatozoa selected with Bovicoll. Mini-SLC (single-layer centrifugation) and swim-up (Control) were performed and cleavage rate and blastocyst rate did not differ significantly between groups. As there was a trend to an increased number of cells in blastocysts in the SLC group, the Mini-SLC method is at least as good as swim-up for selecting frozen-thawed bull spermatozoa for in vitro fertilization (IVF). This method could potentially be used to prepare human sperm for assisted reproduction.
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- Al-Essawe, Essraa M, et al.
(författare)
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Extenders for alpaca epididymal spermatozoa: Comparison of INRA96 and andromed
- 2020
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Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 223
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Tidskriftsartikel (refereegranskat)abstract
- Artificial insemination would be a useful technique for alpaca breeders to use as an aid to breeding to increase fleece quality. The technique, however, is not well developed in alpacas, partly because of the viscous nature of their seminal plasma. Castration conducted for husbandry purposes can provide a source of epididymal spermatozoa to test semen extenders or handling regimens, thus circumventing the problem of the viscous ejaculate. In this experiment, two semen extenders (Andromed and INRA96) developed for other species (bovine and equine, respectively) were tested with alpaca spermatozoa derived from the cauda epididymis. Sperm total motility (mean ± SEM A: 29.1 ± 4.8 % compared with I: 35.4 ± 4.8 %; NS), membrane integrity (A: 58 ± 9% compared with I: 56 ± 9%; NS) and acrosome integrity (A: 65 ± 7% compared with I: 54 ± 7%; NS) were not different between the two extenders. Progressive motility with use of INRA96 was greater after incubating for 30 min than after incubating for 10 min (35 ± 4% vs. 12 ± 4%, respectively; P = 0.03). In conclusion, viable epididymal spermatozoa could be extracted from the castrated organs after overnight transport. There were no differences in sperm quality between the two extenders; therefore, it appears that either extender could be used for alpaca spermatozoa. These results could help in the development of a technique for artificial insemination in alpacas.
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- Cojkic, Aleksandar, et al.
(författare)
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Single layer centrifugation as a method for bacterial reduction in bull semen for assisted reproduction
- 2024
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Ingår i: Veterinary Research Communications. - 0165-7380 .- 1573-7446. ; 48, s. 39-48
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Tidskriftsartikel (refereegranskat)abstract
- Semen samples contain bacteria originating from the animal urogenital tract, environment, and/or contamination during semen processing, negatively affecting sperm quality by producing toxins and/or competing for nutrients in extenders. The aims of this study were to evaluate two methods of Single-layer centrifuges (SLC), high and low density colloid, as a method for bacterial removal from bull semen, and to evaluate sperm quality after treatment. In total, semen samples from 20 bulls (3 ejaculates per bull) were used in this study. Bacterial reduction was evaluated by bacterial quantification (colony forming unit - CFU/mL) while bacterial identification was performed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) after culturing bacteria on blood agar. Sperm motility parameters were evaluated by Computer Assisted Sperm Analyses (CASA), and sperm chromatin structure assay (SCSA) by Flow cytometry. Both, High and Low density SLC reduced number of bacteria significantly (p < 0.001) compared with control. The difference in bacterial count between High and Low SLC was also significant (p < 0.001). Furthermore, High density SLC was successful in removing almost all Bacillus and Proteus spp. Most CASA parameters were significantly improved after both treatments (p < 0.001, p < 0.01, p < 0.05). The Deoxyribonucleic acid (DNA) fragmentation index evaluated by SCSA in High (p < 0.01) and Low (p < 0.05) SLC group differed significantly compared with control. Single-layer centrifugation (SLC) with either a high or a low density colloid is a suitable method for bacterial removal in bull semen.
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- Deori, Sourabh, et al.
(författare)
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Single Layer Centrifugation with 20% or 30% Porcicoll separates the majority of spermatozoa from a sample without adversely affecting sperm quality
- 2020
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Ingår i: Reproduction in Domestic Animals. - : Wiley. - 0936-6768 .- 1439-0531. ; 55, s. 1337-1342
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Tidskriftsartikel (refereegranskat)abstract
- Centrifugation of boar semen through one layer of 40% colloid (Porcicoll) was previously shown to separate spermatozoa from bacteria without having a detrimental effect on sperm quality. However, some spermatozoa were lost. The purpose of the present study was to determine whether 20% or 30% Porcicoll could be used to recover most of the spermatozoa without impacting on sperm quality. Insemination doses (n = 10) from a commercial boar station were sent to the laboratory at the Swedish University of Agricultural Sciences and processed by Single Layer Centrifugation with 20% and 30% Porcicoll approximately 7 hr after semen collection. The resulting sperm samples and controls were evaluated for sperm quality immediately and again after storage at 16-18 degrees C for 4 and 7 days. Sperm recovery was 94 +/- 18% and 87 +/- 15% for 20% and 30% Porcicoll, respectively (p > .05). Sperm mitochondrial membrane potential and chromatin integrity were unaffected (p > .05). The proportion of live spermatozoa producing superoxide (9 +/- 8%, 7 +/- 6% and 3 +/- 1%;p < .05), and the proportion of spermatozoa with high stainability DNA (0.68 +/- 19%, 0.61 +/- 0.22% and 0.96 +/- 0.23%;p < .05- <0.01), were marginally increased whereas membrane integrity, although high, was lower in the centrifuged samples than in the controls (82 +/- 8%, 83 +/- 5% versus 92 +/- 4%;p < .05). In conclusion, centrifugation through 20% or 30% Porcicoll enables most spermatozoa to be recovered, without having a major effect on sperm quality. These results are encouraging for further studies involving microbiological investigation of the processed samples, and scaling-up to process larger volumes of boar ejaculates.
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- Goodla, Lavanya, et al.
(författare)
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Quality of bull spermatozoa after preparation by single-layer centrifugation
- 2014
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Ingår i: Journal of Dairy Science. - : American Dairy Science Association. - 0022-0302 .- 1525-3198. ; 97, s. 2204-2212
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Tidskriftsartikel (refereegranskat)abstract
- The present study aimed to evaluate the effect of single-layer centrifugation (SLC) through a species-specific colloid (Androcoll-B; patent pending, J. M. Morrell) on bull sperm quality. Computer-assisted sperm analysis of motility and flow cytometric analysis of sperm viability (SYBR-14/propidium iodide staining), chromatin integrity (acridine orange staining), reactive oxygen species production [Hoechst 33258-hydroethidine-2',7'-dichlorodihydrofluorescein diacetate (HO-HE-DCFDA) staining], mitochondrial membrane potential (staining with JC-1 probe), and protein tyrosine phosphorylation (specific antibody staining) were performed on unselected and SLC-selected sperm samples. Single-layer centrifugation of bull spermatozoa resulted in the selection of a sperm population that had high mitochondrial membrane potential, a higher content of phosphorylated protein, and more reactive oxygen species than control samples. Sperm chromatin damage was lower in the SLC samples although sperm viability and motility did not differ between SLC samples and controls. These observations suggest that SLC of bull semen in a soybean-containing extender improved some, but not all, parameters of sperm quality.
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- Johannisson, Anders, et al.
(författare)
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Simultaneous evaluation of superoxide content and mitochondrial membrane potential in stallion semen samples provides additional information about sperm quality
- 2018
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Ingår i: Animal Reproduction Science. - : Elsevier BV. - 0378-4320 .- 1873-2232. ; 192, s. 290-297
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Tidskriftsartikel (refereegranskat)abstract
- An improved fertility prediction for stallions is of importance for equine breeding. Here, we investigate the potential of a combined staining of stallion spermatozoa for superoxide and mitochondrial membrane potential (MMP) for this purpose. Semen samples were analysed immediately after arrival at the laboratory, as well as after 24 h. Superoxide was measured by MitoSOXRed, while MMP was measured with JC-1. Menadione was used to stimulate superoxide production. In addition, other parameters of sperm quality, namely motility, membrane integrity, chromatin integrity, sperm kinematics and Hoechst 33258 exclusion were measured and correlated to superoxide production and MMP. Both bivariate correlations between measured parameters as well as multivariate analysis were performed. Measured values in the superoxide/MMP assay did not correlate with other parameters. However, there was a strong negative correlation (r = 0.96 after 0 h, r = 0.95 after 24 h) between membrane integrity and chromatin integrity. Moderate positive correlations were found between motility parameters and membrane integrity, as well as moderate negative correlations between motility parameters and chromatin integrity. The multivariate analysis revealed that membrane integrity, chromatin integrity and motility contributed to the first principal component, while the second was influenced by superoxide/MMP parameters as well as sperm kinematics. Storage of samples for 24 h decreased motility, chromatin integrity and membrane integrity. In conclusion, combined measurement of superoxide and MMP provides additional information not obtained by other assays of sperm quality.
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