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- Nagy, Szabolcs, et al.
(författare)
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Evaluation of domestic animal sperm head morphology via flow cytometric DNA labelling and pulse shape analysis using bull and stallion spermatozoa as model species
- 2023
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Ingår i: Reproduction in domestic animals. - : John Wiley & Sons. - 0936-6768 .- 1439-0531. ; 58:11, s. 1569-1575
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Tidskriftsartikel (refereegranskat)abstract
- The aim of the present study was to test a rapid, robust flow cytometric technique for the detection of sperm head abnormalities of domestic bulls and stallions. The so-called PulSA approach detects the pulse profiles of propidium-iodide labelled spermatozoa. In the first experiment, species-specific threshold values were established on sperm samples that were tested for sperm head abnormalities with a classic visual morphology analysis. In the second experiment, serial mixtures of bull and stallion spermatozoa mimicking different percentages of sperm head abnormalities were analysed. Non-metric multidimensional scaling showed a clear separation between the normal and mixed samples. The PulSA approach may be a useful tool in identifying sub- or infertile breeding males as well as in studying the evolutionary aspects of sperm morphology and morphometry.
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- Valeanu, Sabina, et al.
(författare)
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Seasonal variation in sperm quality parameters in Swedish red dairy bulls used for artificial insemination
- 2015
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Ingår i: Livestock Science. - : Elsevier BV. - 1871-1413 .- 1878-0490. ; 173, s. 111-118
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Tidskriftsartikel (refereegranskat)abstract
- The aim of the present study was to investigate the extent to which seasonal factors such as temperature, atmospheric pressure or visible light length affect dairy bull sperm quality. Straws from 10 dairy bulls, from semen collections in three different seasons, were available for this study. The following quality parameters were assessed: motility, measured by computer assisted sperm motility analysis (SpermVision motility analyzer), membrane integrity (staining with SYBR14/PI (Propidium lodide)with flow cytometric measurement of fluorescence), content of reactive oxygen species (ROS) measured using HE(Hydroethidine) and DCFDA (2', 7'-dichlorodihydrofluorescein diacetate), mitochondrial membrane potential (using JC-1 fluorescence stain), acrosome reaction (measured with fluorescein isothiocyanate-PNA (FITC-PNA) labeling combined with calcium ionophore A23187 and PI), DNA fragmentation index (%DFI, Sperm Chromatin Structure Assay) as well as morphology, using the William's staining protocol. Data was analyzed by analysis of variance (PROC MIXED), using SAS software. The proportion (%) (mean +/- SD) of living, dying and dead sperm cells varied between seasons, with the proportion of living spermatozoa being lowest in summer, although a significant difference (P < 0.05) was only observed between spring and summer. The %DFI assessed by SCSA was lowest in spring and differed significantly (p < 0.04) from summer. A trend towards significance was observed between spring and summer for the motility parameters linearity (LIN) (P=0.068) and straightness (STR) (P=0.062). (C) 2014 Elsevier B.V. All rights reserved.
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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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- 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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