Sperm Preservation and Sexing Technologies in Large- and Small-Ruminant Reproduction: Current Status and Future Perspectives

A special issue of Animals (ISSN 2076-2615). This special issue belongs to the section "Animal Reproduction".

Deadline for manuscript submissions: 15 February 2026 | Viewed by 1527

Special Issue Editors


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Guest Editor
Department of Animal Sciences, Faculty of Agrobiology, Food, and Natural Resources, Czech University of Life Sciences Prague, 165 00 Prague, Czech Republic
Interests: ram and buck sperm cryopreservation; ram and buck semen analysis; flow cytometry of sperm from large and small ruminants; sperm motility evaluation; semen diluents for small ruminants; artificial insemination of sheep and goats
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E-Mail Website
Guest Editor
Department of Animal Sciences, Faculty of Agrobiology, Food, and Natural Resources, Czech University of Life Sciences Prague, 165 00 Prague, Czech Republic
Interests: breeding of small ruminants; artificial insemination; sheep and goats synchronization; sheep husbandry; goats husbandry; animal biotechnology; sheep production; sperm cryopreservation
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

This Special Issue plans to provide an overview of the most recent advances in the field of sperm preservation and sexing technologies used in large- and small-ruminant reproduction.

This Special Issue aims to provide selected contributions on topics including, but not limited to, the following:

  • Optimization of insemination dose processing;
  • In vitro assays that predict the fertilizing ability of insemination doses;
  • Ethical questions related to the artificial insemination of large and small ruminants;
  • Development of sperm diluents for fresh, chilled, or cryopreserved insemination doses;
  • Technologies of sperm preservation (especially alternative techniques for sperm preservation without liquid nitrogen);
  • Technologies of sperm sexing (especially alternatives to the flow-cytometry-based method);
  • Optimizing artificial insemination methods (especially those appropriate for small ruminants);
  • Hormonal and non-hormonal synchronization schemes.

Dr. Filipp Georgijevič Savvulidi
Dr. Martin Ptáček
Guest Editors

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Keywords

  • semen
  • cryopreservation
  • liquid storage
  • sperm lyophilization
  • sexed semen
  • sperm quality
  • biotech-nology
  • domestic and wild ruminants

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Published Papers (2 papers)

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Research

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19 pages, 8767 KB  
Article
Lipidomic Landscapes of Cryopreserved Sperm from Alpine and Spanish–Creole Bucks
by Mustafa Bodu, Mustafa Hitit, Selamawit Woldesenbet, Muhammet Raşit Uğur, Zeynep Erdoğan, Olivia Chika Greenwood, Raheem Davian Murray, Andres Pech Cervantes and Erdoğan Memili
Animals 2025, 15(13), 1897; https://doi.org/10.3390/ani15131897 - 27 Jun 2025
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Abstract
This study provides a comparative analysis of the post-thaw sperm lipidomic profiles of Alpine and Spanish–Creole goat breeds to explore breed-specific differences in fatty acid composition and their implications for sperm function and reproductive efficiency. Lipids were extracted from cryopreserved semen samples of [...] Read more.
This study provides a comparative analysis of the post-thaw sperm lipidomic profiles of Alpine and Spanish–Creole goat breeds to explore breed-specific differences in fatty acid composition and their implications for sperm function and reproductive efficiency. Lipids were extracted from cryopreserved semen samples of Alpine (n = 7) and Spanish–Creole (n = 4) mature bucks and subsequently analyzed by gas chromatography–mass spectrometry (GC-MS), with 21 fatty acids identified within the two breeds. Eight of these fatty acids, namely 13:0, 16:0, 18:0, 24:0, 14:1, 18:1 (cis-9), 24:1, and 18:2 showed significant differences (p < 0.05). The levels of 16:0, 18:0, 24:0, 18:1 (cis-9), 18:1, and 18:2 were higher in the Alpine breed, whereas the levels of 13:0, 14:1, and 24:1 were higher in the Spanish–Creole breed (p < 0.05). Of those, 16:0, 18:1 (cis-9), and 18:2 were both statistically and biologically significant (p < 0.05, FC > 2). Concentrations of the total fatty acids, total saturated fatty acids (Total-SFA), and total polyunsaturated fatty acids (Total-PUFA) were significantly higher in the Alpine breed, whereas the concentrations of the total cis-monounsaturated fatty acid (Total cis-MUFA) were significantly higher in the Spanish–Creole breed (p < 0.05). Network and pathway analyses revealed that 16:0, 18:1 (cis-9), and 18:2 contributed to the most central nodes of the lipidomic network, which may support membrane stability and cryotolerance. The lipidomic differences observed between breeds may be attributed to both genetic and environmental factors and may provide valuable tools for enhancing breeding strategies, artificial insemination programs, and sperm cryopreservation techniques. Full article
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Review

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14 pages, 700 KB  
Review
Molecular Constraints of Sperm Sex Sorting via TLR7/8 Activation
by Alikhan Magauiya, Kausar Torebek, Filipp Georgijevič Savvulidi, Martin Ptáček, Christopher LeBrun, Lucie Langerová, Elbosyn Sagdat, Saltanat Baikoshkarova and Nurlan Malmakov
Animals 2025, 15(20), 2976; https://doi.org/10.3390/ani15202976 - 14 Oct 2025
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Abstract
In modern livestock, the demand for sperm sex selection technologies is high, as the ability to deliberately produce offspring of a specific sex offers significant economic advantages. Traditionally, sperm sorting is performed using Fluorescence-Activated Cell Sorting. However, the flow cytometric method is expensive, [...] Read more.
In modern livestock, the demand for sperm sex selection technologies is high, as the ability to deliberately produce offspring of a specific sex offers significant economic advantages. Traditionally, sperm sorting is performed using Fluorescence-Activated Cell Sorting. However, the flow cytometric method is expensive, technically complex, and associated with reduced sperm viability. An alternative promising method involves the use of Toll-like TLR7/8 receptors for the selective binding of spermatozoa of a particular sex. It was discovered previously that the activation of TLR7/8 by its ligand(s) selectively inhibits the motility of X-bearing sperm without affecting the motility of Y-bearing sperm. The swim-up technique, which separates sperm based on sex chromosome type by isolating fractions enriched in either X- or Y-bearing gametes due to differences in their motility, can be used with this method. Sperm sex sorting via the TLR7/8 activation is cheap, technically non-complex, and does not affect sperm viability negatively. The goal of this review is to provide an overview of the TLR7/8-dependent sperm sorting method. Further, we discuss why the method of sperm sorting via TLR7/8 activation is successfully implemented in some animal species (such as murine, caprine, ovine, and bovine) but fails in others, like swine and canine. Full article
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