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Advances in Animal Fertility Preservation—Second Edition
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Advances in Animal Fertility Preservation—Second Edition

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

Deadline for manuscript submissions: closed (31 August 2024) | Viewed by 8958

Special Issue Editor

Dr. Rosa Maria Lino Neto Pereira

E-Mail Website
Guest Editor
1. Unit of Biotechnology and Genetic Resources, National Institute of Agrarian and Veterinary Research, Quinta da Fonte Boa, 2005-048 Vale de Santarém, Portugal
2. Department of Veterinary Sciences Research Centre, Vasco da Gama University School, Lordemão University Campus, 3020-210 Coimbra, Portugal
3. CIISA, Faculty of Veterinary Medicine, University of Lisbon, Av. da Universidade Técnica, 1300-477 Lisboa, Portugal
Interests: physiology; reproduction; assisted reproductive technologies; animal genetic resources characterization and conservation; gametes and embryo metabolism; cryobiology
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The preservation and sustainable management of animal genetic resources are critical for the global maintenance of biodiversity, food security, and population livelihoods. A concerted effort, including the establishment of international and national goals and strategies allied to local interventions, is urgently needed for the conservation of animal genetic diversity. Currently, most of the indicators of the state of biodiversity are in decline. This decline is global and is not slowing down. On the contrary, the extinction of species and breeds, including domestic animals, continues to increase. Germplasm preservation and assisted reproductive technologies are currently envisaged as critical tools for the conservation and management of animal genetic resources and for fertility preservation. However, germplasm quality is one of the key limiting factors in both male and female fertility. This Special Issue presents an excellent opportunity to show novel strategies, therapies, and lines of research in progress that have revealed different possibilities in the conservation of animal genetic resources, the preservation of germplasm, and the improvement in the quality and developmental competence of gametes and embryos.

Dr. Rosa Maria L. N. Pereira
Guest Editor

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Keywords

  • animal genetic resources conservation
  • assisted reproductive technologies
  • germplasm
  • cryopreservation
  • development
  • pregnancy

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Published Papers (6 papers)

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Research

24 pages, 3112 KiB  
Article
Effect of Seminal Plasma on the Freezability of Boar Sperm
by Kuanfeng Zhu, Yukun Song, Zhi He, Peng Wang, Xuguang Wang and Guoshi Liu
Animals 2024, 14(24), 3656; https://doi.org/10.3390/ani14243656 - 18 Dec 2024
Cited by 1 | Viewed by 999
Abstract
Background: Seminal plasma is an important component of semen and has a significant effect on sperm function. However, the relationship between seminal plasma and sperm freezing capacity has not been fully studied. Purpose: Exploring metabolites and proteins related to the boar sperm freezing [...] Read more.
Background: Seminal plasma is an important component of semen and has a significant effect on sperm function. However, the relationship between seminal plasma and sperm freezing capacity has not been fully studied. Purpose: Exploring metabolites and proteins related to the boar sperm freezing capacity in seminal plasma, by metabolomic and proteomic approaches, and directly verifying the protective effect of seminal plasma on the cryopreservation of boar sperm using high and low freezability seminal plasma as base freezing extender. Methods: Semen samples were collected from 30 different boars, 11 high and 11 low freezing-resistant boars were selected after freezing 2~4 times, and seminal plasma was selected at the same time. Sperm motility and movement parameters were analyzed using a CASA system. Reproductive hormones (Testosterone, progesterone, estradiol, prolactin, prostaglandin F2α, luteinoid hormone) in seminal plasma were detected by ELISA. Analysis of proteins and metabolites in high and low freezing-resistant seminal plasma by proteomics and metabolomics techniques. Results: The six reproductive hormones tested were not significantly associated with sperm freezing resistance. A total of 13 differentially expressed metabolites (DEMs) and 38 differentially expressed proteins (DEPs) were identified, while a total of 348 metabolites and 1000 proteins were identified. These DEMs were related to energy metabolism, drugs, or environmental pollutants, while the DEPs were mainly involved in the cytoskeletal dynamics and cell adhesion processes. There were 33 metabolites and 70 proteins significantly associated with mean progress motility (PM) at 10 min and 2 h after thawing. The 70 related proteins were associated with cell division and cycle regulation in gene ontology (GO) terms, as well as KEGG pathways, thermogeneration, and pyruvate metabolism. Using highly freezable boar SP as a base freezing extender made no difference from using lowly freezable boar SP, and both were not as good as the commercial control. Conclusion: There were significant differences in seminal plasma with different freezability, but the similarity was much greater than the difference. The protection effect of seminal plasma is not remarkable, and it does not exhibit superior cryoprotective properties compared to commercial semen cryoelongators. Significance: This study provides a deeper understanding of how seminal plasma composition affects sperm freezabilty. It provides potential biomarkers and targets for improving sperm cryopreservation techniques. Full article
(This article belongs to the Special Issue Advances in Animal Fertility Preservation—Second Edition)
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8 pages, 4279 KiB  
Communication
Development of Germ Cell Isolation and Optimal Cryopreservation Method for Lissachatina fulica (L. fulica)
by Jukyeong Jeong, Seungki Lee and Jung Kyu Choi
Animals 2024, 14(22), 3229; https://doi.org/10.3390/ani14223229 - 11 Nov 2024
Viewed by 777
Abstract
This study aims to develop an optimized method for cryopreserving the germ cells of Lissachatina fulica (L. fulica) using vitrification, as an alternative approach for conserving endangered snail species. First, we isolated several key reproductive organs, including the sperm oviduct, albumen [...] Read more.
This study aims to develop an optimized method for cryopreserving the germ cells of Lissachatina fulica (L. fulica) using vitrification, as an alternative approach for conserving endangered snail species. First, we isolated several key reproductive organs, including the sperm oviduct, albumen gland, hermaphrodite gland (ovotestis), and hermaphrodite duct from L. fulica. When the ovotestis was finely chopped, numerous sperm with long tails and distinct heads were observed. The staining of sperm nuclei was confirmed using Hoechst 33342 dye. Since the hermaphrodite gland, referred to as the ovotestis, contains both male and female germ cells, we performed tissue staining on the ovotestis using hematoxylin and eosin (H&E) dye. H&E staining of the ovotestis revealed numerous oval-shaped acini containing sperm and early germ cells. Spermatocytes and spermatids were observed within distinct boundaries, with mature sperm appearing following spermatogenesis. To preserve the species of the L. fulica, we introduced vitrification technology to cryopreserve its reproductive organs. The non-vitrification group showed an average cell viability of 96.6%, while the vitrification group had 86.8% after thawing. This study presents a reliable cryopreservation protocol for L. fulica, with potential applications for other endangered snails, supporting conservation efforts to preserve genetic resources and biodiversity. Full article
(This article belongs to the Special Issue Advances in Animal Fertility Preservation—Second Edition)
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15 pages, 1745 KiB  
Article
Effect of Urolithin A on Bovine Sperm Capacitation and In Vitro Fertilization
by Manuela Jorge, Filipa C. Ferreira, Carla C. Marques, Maria C. Batista, Paulo J. Oliveira, F. Lidon, Sofia C. Duarte, José Teixeira and Rosa M. L. N. Pereira
Animals 2024, 14(18), 2726; https://doi.org/10.3390/ani14182726 - 20 Sep 2024
Cited by 2 | Viewed by 1397
Abstract
Reactive oxygen species (ROS) play a critical role in the functional competence of sperm cells. Conversely, excessive generation of ROS can impair sperm function, including their fertilization ability. Urolithin A (UA), a gut bacteria-derived metabolite produced from the transformation of ellagitannins, with anti-aging [...] Read more.
Reactive oxygen species (ROS) play a critical role in the functional competence of sperm cells. Conversely, excessive generation of ROS can impair sperm function, including their fertilization ability. Urolithin A (UA), a gut bacteria-derived metabolite produced from the transformation of ellagitannins, with anti-aging and antioxidant properties, was investigated for the first time in bovine sperm cells in the present study. Firstly, different doses of UA (0, 1, and 10 μM; 8–16 sessions) were used during the capacitation process of frozen-thawed bovine sperm. Sperm motility was assessed using optical microscopy and CASA. Sperm vitality (eosin-nigrosin), ROS, and ATP levels, as well as mitochondrial membrane potential (JC1) and oxygen consumption were evaluated. A second experiment to test the effect of different doses of UA (0, 1, and 10 μM; 9 sessions) in both the capacitation medium, as above, and the fertilization medium, was also implemented. The embryonic development and quality were evaluated. UA, at a concentration of 1 μM, significantly improved sperm movement quality (p < 0.03). There was a trend towards an increase in the oxygen consumption rate (OCR) of capacitated sperm with 1 μM and 10 μM UA supplementation. Moreover, an increase in ATP levels (p < 0.01) was observed, accompanied by a reduction in ROS levels at the higher UA concentration. These results suggest that UA may enhance spermatozoa mitochondrial function, modifying their metabolic activity while reducing the oxidative stress. Also, the number of produced embryos appears to be positively affected by UA supplementation, although differences between the bulls may have mitigated this effect. In conclusion, presented results further support previous findings indicating the potential therapeutic value of UA for addressing reproductive sub/infertility problems and improving ART outcomes. In addition, our results also reinforce the important bull effect on ART and that male sperm bioenergetic parameters should be used to predict spermatozoa functionality and developmental potential. Full article
(This article belongs to the Special Issue Advances in Animal Fertility Preservation—Second Edition)
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16 pages, 4502 KiB  
Article
Effects of Cryoprotectant Concentration and Exposure Time during Vitrification of Immature Pre-Pubertal Lamb Cumulus–Oocyte Complexes on Nuclear and Cytoplasmic Maturation
by Letizia Temerario, Nicola Antonio Martino, Monika Bennink, Agnes de Wit, Sipke Joost Hiemstra, Maria Elena Dell’Aquila and Julie Lamy
Animals 2024, 14(16), 2351; https://doi.org/10.3390/ani14162351 - 14 Aug 2024
Cited by 2 | Viewed by 1243
Abstract
Oocyte vitrification allows for the storing of endangered breed female gametes. Cryoprotectant (CPA) concentration and exposure time should ensure cell protection with minimal toxicity. In the present study, a high concentration-rapid exposure (HC-RE) and a low concentration-slow exposure (LC-SE) vitrification protocol, using dimethyl [...] Read more.
Oocyte vitrification allows for the storing of endangered breed female gametes. Cryoprotectant (CPA) concentration and exposure time should ensure cell protection with minimal toxicity. In the present study, a high concentration-rapid exposure (HC-RE) and a low concentration-slow exposure (LC-SE) vitrification protocol, using dimethyl sulfoxide (DMSO) and ethylene glycol (EG) as permeating CPAs, were evaluated on meiotic competence and bioenergetic-oxidative status of pre-pubertal lamb immature COCs after in vitro maturation (IVM). For each protocol, COCs vitrified through a traditional protocol and fresh ones were used as controls. Both protocols allowed COC morphology preservation after vitrification-warming (V-W) and cumulus expansion after IVM. The maturation rate (7% and 14%) was comparable to the vitrified control (13% and 21%) but not satisfactory compared to fresh ones (58% and 64%; p < 0.001). The rate of mature oocytes displaying a perinuclear/subcortical (P/S) mitochondrial distribution pattern, an index of cytoplasmic maturity, was comparable between vitrified and fresh oocytes. The LC-SE vitrification protocol did not affect quantitative bioenergetic-oxidative parameters compared to both controls whereas HC-RE protocol significantly reduced intracellular reactive oxygen species (ROS) levels, indicating cell viability loss. In conclusion, to improve pre-pubertal lamb immature COC vitrification, the combination of low CPA concentrations with prolonged exposure time could be more promising to investigate further. Full article
(This article belongs to the Special Issue Advances in Animal Fertility Preservation—Second Edition)
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14 pages, 1556 KiB  
Article
The Influence of Selected Factors on Changes in Locomotion Activity during Estrus in Dairy Cows
by Mária Mičiaková, Peter Strapák and Eva Strapáková
Animals 2024, 14(10), 1421; https://doi.org/10.3390/ani14101421 - 9 May 2024
Cited by 2 | Viewed by 1574
Abstract
The objective of this study was the evaluation of the locomotion activity of heifers and Holstein dairy cows during estrus. We have analyzed the locomotion activity using the Heatime RuminAct device on 180 cows (32 heifers and 148 dairy cows) and we evaluated [...] Read more.
The objective of this study was the evaluation of the locomotion activity of heifers and Holstein dairy cows during estrus. We have analyzed the locomotion activity using the Heatime RuminAct device on 180 cows (32 heifers and 148 dairy cows) and we evaluated a total of 633 estrus cycles during the reference period of 3 days before estrus, 3 days after estrus, and on the day ofestrus occurrence. The datawere analyzed using the DataFlowTM II program. The locomotion of cows was expressed in the units of locomotion activity in 24 h (u.24 h−1). During the reference period of 3 days before estrus, the cows showed locomotion activity of 558 u.24 h−1, with an increase in locomotion activity on the day of estrus of 836 u.24 h−1, and, during the reference period of 3 days after estrus, the level of locomotion activity decreased to 537 836 u.24 h−1, which is a similar level of locomotion activity to the reference period before estrus. Through the statistical analysis, we evaluated the impact of parity, lactation stage, milk yield, and individuality on changes in locomotion activity during estrus and throughout the reference period, and we found a significant effect of parity (F = 13.41, p < 0.001) on changes in the locomotion activity of dairy cows during estrus. Based on these results, this research offers fresh perspectives on assessing specific factors affecting the locomotion activity of dairy cows during estrus through the practical application of electronic systems for estrus detection on dairy farms. Full article
(This article belongs to the Special Issue Advances in Animal Fertility Preservation—Second Edition)
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13 pages, 4925 KiB  
Article
Fertility and Insemination Characteristics of Sperm Storage Tubules in Old Thai-Native Hens
by Theerapat Kheawkanha, Vibuntita Chankitisakul, Maruay Pimprasert, Wuttigrai Boonkum and Thevin Vongpralub
Animals 2024, 14(5), 694; https://doi.org/10.3390/ani14050694 - 23 Feb 2024
Cited by 2 | Viewed by 2154
Abstract
We aimed to evaluate the effects of sperm concentration (150–250 × 106 spz/dose) and insemination frequency (once, twice, and thrice weekly) on fertility and sperm storage tubule (SST) characteristics. The SSTs were classified into five categories: namely, SSTs having an unscorable (SST1), [...] Read more.
We aimed to evaluate the effects of sperm concentration (150–250 × 106 spz/dose) and insemination frequency (once, twice, and thrice weekly) on fertility and sperm storage tubule (SST) characteristics. The SSTs were classified into five categories: namely, SSTs having an unscorable (SST1), empty (SST2), low (SST3), medium (SST4), and high (SST5) sperm count after insemination. The results showed that only insemination frequency affected the fertility rate (p < 0.05). The highest fertility was found in the thrice-weekly insemination group; however, this rate was not significantly different from that for the twice-weekly insemination group, except on day 7, while the once-weekly insemination group showed the lowest fertility rate (p < 0.05) from day four onward. On day 1, the SST characteristics showed no differences among the various insemination frequencies. On day 4, the SST2 and SST3 categories increased in the once-weekly insemination group (p < 0.05), while the SST4 and SST5 categories decreased compared to the twice- and thrice-weekly insemination groups (p < 0.05). On day 7, only the thrice-weekly insemination group maintained a level of SST5 category tubules like that measured on day 1 (p > 0.05). In summary, the insemination dose of 150 × 106 sperm was enough for fertilization, and thrice-weekly insemination was the appropriate frequency in old Thai native hens for maintaining a high sperm density in the SSTs throughout the week. Full article
(This article belongs to the Special Issue Advances in Animal Fertility Preservation—Second Edition)
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