Advances in Animal Sperm Conservation Techniques for Better Fertility—2nd Edition

A special issue of Veterinary Sciences (ISSN 2306-7381). This special issue belongs to the section "Veterinary Reproduction and Obstetrics".

Deadline for manuscript submissions: 31 July 2025 | Viewed by 1361

Special Issue Editors


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Guest Editor
Department of Veterinary Medicine, University of Milan, Via dell’Università 6, 26900 Lodi, Italy
Interests: local poultry breeds; poultry semen cryopreservation; poultry reproduction
Special Issues, Collections and Topics in MDPI journals

E-Mail Website
Guest Editor
Department of Veterinary Medicine and Animal Science, University of Milan, 26900 Lodi, Italy
Interests: animal science and technology; pig nutrition; poultry nutrition; microbiome; feed additives; gut health
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Sperm conservation is one of the most important biotechnologies for the advancement of assisted reproduction in domestic and wild animals. Sperm conservation is essential for the breeding management of endangered breeds or species, for genetic improvement in livestock breeding, to prevent the loss of animal genetic variability and to solve infertility problems in high-quality individuals.

Despite the large amount of progress that has been made in this field, the cellular mechanisms involved in sperm conservation have not yet been entirely explained. In particular, the fertility values recorded with the use of frozen/thawed sperm are still low in different animal species. New knowledge and investigations to improve the quality and fertilizing ability of stored semen in various species are, therefore, essential.

We are pleased to invite you to contribute to this Special Issue, which aims to publish original research articles or reviews on “Advances in Animal Sperm Conservation Techniques for Better Fertility”.

We invite researchers to contribute their recent studies, focusing principally, but not exhaustively, on the following topics:

  • Semen extenders;
  • Recent discoveries on the use of new cellular cryoprotectants;
  • Novel cryopreservation procedures;
  • Predictive markers for sperm fertility;

Technologies for the assessment of sperm integrity and functions.

Prof. Dr. Silvia Cerolini
Dr. Vera Perricone
Guest Editors

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Veterinary Sciences is an international peer-reviewed open access monthly journal published by MDPI.

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Keywords

  • spermatozoa
  • fertility
  • sperm conservation
  • cryoprotectant
  • semen extender
  • semen evaluation

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Related Special Issue

Published Papers (3 papers)

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Research

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13 pages, 563 KiB  
Article
Validating Sperm Concentration in Rabbit Cryopreservation Protocol: Implications for Fertility, Litter Size, and Offspring Growth
by Michele Di Iorio, Giusy Rusco, Fabrizio Lauriola, Emanuele Antenucci, Alessandra Roncarati, Silvia Cerolini, Michele Schiavitto and Nicolaia Iaffaldano
Vet. Sci. 2025, 12(7), 678; https://doi.org/10.3390/vetsci12070678 - 18 Jul 2025
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Abstract
The cryopreservation of rabbit semen is a valuable strategy for genetic resource preservation and efficient artificial insemination, but outcomes remain inconsistent, partly due to variations in sperm concentration per dose. This study aimed to evaluate the in vivo effects of different sperm concentrations [...] Read more.
The cryopreservation of rabbit semen is a valuable strategy for genetic resource preservation and efficient artificial insemination, but outcomes remain inconsistent, partly due to variations in sperm concentration per dose. This study aimed to evaluate the in vivo effects of different sperm concentrations (15, 25, 35, 55, and 75 million per straw) on fertility, prolificacy, and offspring growth in nulliparous and multiparous does. A total of 384 rabbit females were inseminated using frozen–thawed semen, and their reproductive performance was compared with fresh semen. Fertility and kindling rates varied with sperm concentration and parity: nulliparous does showed the highest fertility at 15 million sperm/straw (84.4%), while multiparous does reached peak values at 25–55 million/straw (78.1–81.3%). Litter size and live-born kits were consistently higher in multiparous than in nulliparous does. Offspring body weight at 19 and 60 days was influenced by both sperm concentration and maternal parity, with better growth generally observed in multiparous groups. Weaning success remained high across all groups. Our results indicate that sperm concentrations ranging from 15 to 35 × 106/straw are the most suitable for cryopreservation, as they maintain high fertility, prolificacy, and offspring growth, comparable to fresh semen. These results confirm that optimizing sperm concentration during cryopreservation improves reproductive efficiency and that tailoring insemination strategies to the physiological status of the female enhances outcomes. The results provide useful recommendations for improving cryopreservation techniques in rabbit breeding programs. Full article
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12 pages, 1190 KiB  
Article
Autophagic Degradation of GPX4 Mediates Ferroptosis During Sheep Sperm Cryopreservation
by Boyuan Li, Erhan Hai, Yukun Song and Jiaxin Zhang
Vet. Sci. 2025, 12(5), 490; https://doi.org/10.3390/vetsci12050490 - 19 May 2025
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Abstract
Ferroptosis is implicated in cryodamage to sheep sperm, potentially due to glutathione peroxidase 4 (GPX4) degradation during freezing; however, the pathway underlying GPX4 degradation remains unclear. In this study, a comparison of cryoprotective effects between the autophagy inhibitor chloroquine (CQ) and the ubiquitination [...] Read more.
Ferroptosis is implicated in cryodamage to sheep sperm, potentially due to glutathione peroxidase 4 (GPX4) degradation during freezing; however, the pathway underlying GPX4 degradation remains unclear. In this study, a comparison of cryoprotective effects between the autophagy inhibitor chloroquine (CQ) and the ubiquitination inhibitor MG132 revealed that 5 μM CQ treatment significantly enhanced the motility (p < 0.01) and sperm plasma membrane integrity rate (p < 0.01) of frozen–thawed sperm; no protective effects were observed in any MG132 treatment group. Mechanistic analysis indicated that CQ treatment substantially restored GPX4 protein expression (p < 0.01), and concurrently reduced lipid peroxidation (p < 0.01) and free iron ion accumulation (p < 0.01), in frozen–thawed sperm. These findings suggest that GPX4 degradation during cryopreservation occurs via the autophagy pathway. This study established a ferroptosis–GPX4–autophagy axis during sheep sperm cryopreservation and identified autophagy-mediated GPX4 loss as a potential target for enhancing sperm cryoprotection. Full article
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Review

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15 pages, 1423 KiB  
Review
Sperm Membrane Stability: In-Depth Analysis from Structural Basis to Functional Regulation
by Shan-Hui Xue, Bing-Bing Xu, Xiao-Chun Yan, Jia-Xin Zhang and Rui Su
Vet. Sci. 2025, 12(7), 658; https://doi.org/10.3390/vetsci12070658 - 11 Jul 2025
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Abstract
Sperm membrane stability is a key factor in determining sperm viability and fertilization capability, with broad implications ranging from basic reproductive biology to livestock breeding practices. This comprehensive review examines the structural and functional mechanisms underlying sperm membrane integrity, including defensive barrier functions, [...] Read more.
Sperm membrane stability is a key factor in determining sperm viability and fertilization capability, with broad implications ranging from basic reproductive biology to livestock breeding practices. This comprehensive review examines the structural and functional mechanisms underlying sperm membrane integrity, including defensive barrier functions, potentiometric ion channel regulation, and motility modulation that collectively optimize sperm survival, motility, and fertilization potential. Environmental factors such as temperature fluctuations, abnormal pH levels (outside the optimal 7.2–8.2 range), pathological conditions, and hormonal imbalances can compromise membrane stability by inducing oxidative stress and protein denaturation. Key regulatory proteins, notably NPC2 for cholesterol homeostasis, Flotillin proteins for lipid raft organization, and Annexin V for membrane repair mechanisms, demonstrate essential roles in maintaining structural integrity. In livestock reproduction, membrane stability research facilitates the optimization of cryoprotectant formulations and freezing protocols, resulting in 15–25% improvements in post-thaw sperm survival rates and enhanced artificial insemination success. These findings provide valuable insights for advancing assisted reproductive technologies and improving reproductive efficiency in animal husbandry. Full article
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