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Changeability of sperm chromatin structure during liquid storage of ovine semen in milk-egg yolk- and soybean lecithin-based extenders and their relationships to field-fertility

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Abstract

The aim of this experiment was to study the effect of semen extender on sperm chromatin structure and to correlate chromatin integrity with field-fertility of preserved ram semen. Ejaculates of at least 2 × 109 sperm/ml and 70 % progressive motility were collected using an artificial vagina from Chios rams (n = 11, 4–6 years old), split-diluted to 1 × 109 sperm/ml with milk-egg yolk- and soybean lecithin (Ovixcell®)-based extenders, packaged in 0.5-ml straws and examined after 6, 24 and 48 h of storage at 5 ± 1 °C. Evaluation endpoints were computer-assisted sperm motion analysis, fluorescence-based analysis of chromatin structure by chromomycin A3 and acridine orange assays, and 65-day pregnancy rate (PR) of 34- to 36-h preserved semen after intra-cervical insemination of ewes (n = 154) in progestagen-synchronized estrus. Neither extender nor storage time had any influence on incidence of decondensed chromatin. Unlike Ovixcell® extender, deterioration of sperm motility (P < 0.01) and chromatin stability (P < 0.005) was detected after 48 h of storage in milk-egg yolk extender. Sperm motility accounted for 14.4–18.5 % of variations in chromatin integrity (P < 0.001). No significant difference was found in PR of Ovixcell®- and milk-egg yolk-stored semen. Nevertheless, PR differed between rams (14.3–71.4 %; P < 0.025). Chromatin integrity explained 10.2–56.3 % of variations in PR (P < 0.05–0.01). A pronounced decline in PR (19.1 %) was observed when percentages of decondensed and destabilized chromatin have reached thresholds of 10.5–30 % and 4–9 %, respectively. In conclusion, Ovixcell® is superior to milk-egg yolk extender in preserving chromatin stability and motility. Chromatin defects are negatively associated with sperm fertility.

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Acknowledgments

The authors are indebted to Ms. Angelina Vasiliadou (ATEI of Thessaloniki) for her valued laboratory support and technical assistance. We thank the farm stuff at Thesprotia (Greece) for care of the experimental animals. This work was supported by NAGREF-Veterinary Research Institute, Thessaloniki, Greece.

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The authors have no conflict of interest to declare.

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Authors and Affiliations

  1. EquiBiotech Inc-Research Services in Farm Animal Breeding, 19 Gravias street, 54645, Thessaloniki, Greece

    Tarek Khalifa

  2. Department of Animal Production, ATEI of Thessaloniki, P.O. Box 141, 574 00, Sindos, Thessaloniki, Greece

    Aristotelis Lymberopoulos

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  1. Tarek Khalifa
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Correspondence to Tarek Khalifa.

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Tarek Khalifa and Aristotelis Lymberopoulos contributed equally to the manuscript

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Khalifa, T., Lymberopoulos, A. Changeability of sperm chromatin structure during liquid storage of ovine semen in milk-egg yolk- and soybean lecithin-based extenders and their relationships to field-fertility. Cell Tissue Bank 14, 687–698 (2013). https://doi.org/10.1007/s10561-012-9357-6

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