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  • 1.
    Colleoni, Silvia
    et al.
    AVANTEA.
    Lagutina, Irina
    AVANTEA.
    Lazzari, Giovanna
    AVANTEA.
    Rodriguez-Martinez, Heriberto
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Clinical and Experimental Medicine, Medical Genetics.
    Galli, Cesare
    AVANTEA.
    Morrell, Jane M
    Swedish University Agriculture Science.
    New Methods for Selecting Stallion Spermatozoa for Assisted Reproduction2011In: Journal of Equine Veterinary Science, ISSN 0737-0806, E-ISSN 1542-7412, Vol. 31, no 9, p. 536-541Article in journal (Refereed)
    Abstract [en]

    Improved sperm selection techniques are needed to increase the efficiency of equine-assisted reproduction. Single layer centrifugation (SLC) of spermatozoa has been shown to improve the quality of stallion sperm samples. In this study, the functionality of selected stallion spermatozoa was tested by intracytoplasmic sperm injection of equine oocytes after selection by SLC through Androcoll-E or by discontinuous density gradient centrifugation (DGC) through Redigrad and Tyrodes medium with added albumin, lactate, and pyruvate. The mean cleavage rates of the injected oocytes from SLC- and DGC-selected spermatozoa were 67% and 66%, respectively, whereas the proportion of blastocysts developing from cleaved oocytes was 28% and 22%, respectively (P andgt; .05, not significant). An incidental finding was that there was a tendency for SLC-selected spermatozoa to have a higher percentage of spermatozoa with normal morphology than DGC (70% +/- 22% vs. 58% +/- 38%) and for more blastocysts to be obtained from subfertile ejaculates (21 [19.6%] vs. 15 [14.4%], respectively). In further experiments, stallion spermatozoa bound to hyaluronan, although binding may depend on the semen extender and sperm treatment as well as incubation time. In conclusion, SLC-selected stallion spermatozoa function normally when injected into oocytes. SLC may potentially be better than DGC at selecting spermatozoa from subfertile ejaculates, but this effect needs rigorous investigation with a much larger sample size. Use of the hyaluronan-binding assay for assessing the potential fertility of stallion spermatozoa may be useful but requires further evaluation.

  • 2.
    Morrell, J.M.
    et al.
    Swedish University of Agriculture Science, Sweden .
    Johannisson, A.
    Swedish University of Agriculture Science, Sweden .
    Strutz, H.
    Swedish University of Agriculture Science, Sweden .
    Dalin, A.-M.
    Swedish University of Agriculture Science, Sweden .
    Rodriguez-Martinez, Heriberto
    Swedish University of Agriculture Science, Sweden .
    Colloidal Centrifugation of Stallion Semen: Changes in Sperm Motility, Velocity, and Chromatin Integrity during Storage2009In: Journal of Equine Veterinary Science, ISSN 0737-0806, E-ISSN 1542-7412, Vol. 29, no 1, p. 24-32Article in journal (Refereed)
    Abstract [en]

    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.

  • 3.
    Ortega-Ferrusola, Cristina
    et al.
    University Extremadura.
    Johannisson, Anders
    SLU.
    Pena Vega, Fernando J
    University Extremadura.
    Tapia, Jose A
    University Extremadura.
    Rodriguez-Martinez, Heriberto
    Linköping University, Faculty of Health Sciences. Linköping University, Department of Clinical and Experimental Medicine, Medical Genetics.
    Dalin, Ann M
    SLU.
    Morrell, Jane M
    SLU.
    Effect of Different Extenders and Seminal Plasma on the Susceptibility of Equine Spermatozoa to Lipid Peroxidation After Single-Layer Centrifugation, Through Androcoll-E2011In: Journal of Equine Veterinary Science, ISSN 0737-0806, E-ISSN 1542-7412, Vol. 31, no 7, p. 411-416Article in journal (Refereed)
    Abstract [en]

    his study was conducted in an attempt to see whether single-layer centrifugation (SLC) increases the susceptibility of stallion spermatozoa to lipid peroxidation (LPO), in different extenders after removing all seminal plasma (SP). The susceptibility of stallion spermatozoa to LPO was studied before and after SLC. Each ejaculate was split, and aliquots extended with one of the three different extenders: INRA 96, Kenneys, or Equipro, and stored for 24 hours at 5 degrees C (i). From the extended samples, an aliquot was kept as a control and the other was subjected to SLC through Androcoll-E. The selected spermatozoa were re-suspended in the appropriate extenders, without (ii) or with (iii) addition of 50% (v/v) pooled homologous SP for 24 hours at 5 degrees C. Using ferrous sulfate as pro-oxidant, the susceptibility for LPO was flow-cytometrically assessed using the probe Bodipy(581/591)-C(11). Sperm motility, monitored with a Qualisperm motility analyzer, increased after SLC treatment (P andlt; .001). No significant correlations were found between motility and induced LPO with ferrous sulfate. The SP and extenders, per se, did not have a significant protective effect against LPO, but the interaction between SP and Kenney increased the susceptibility to LPO. However, the selected spermatozoa through Androcoll-E and the subsequent dilution in INRA had a significant protective effect against LPO (P andlt; .05), especially when the oxidative insults were higher (80 mu M).

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