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Animals
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Further Approaches on Sperm Cryopreservation
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Further Approaches on Sperm Cryopreservation

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

Deadline for manuscript submissions: closed (31 January 2022) | Viewed by 14292

Special Issue Editor

Dr. Maria-Jesus Palomo Peiró

E-Mail Website
Guest Editor
Department of Animal Medicine and Surgery, Autonomous University of Barcelona, 08193 Bellaterra, Spain
Interests: small ruminant semen analysis; cryopreservation

Special Issue Information

Dear Colleagues,

Sperm cryopreservation has been a useful tool for the storage and restoration of valuable genetic resources in combination with assisted reproductive techniques This semen technology has become indispensable for genetic improvement toward breeding management in the livestock industry, for the preservation of endangered breeds or species toward preventing the loss of animal genetic variability or for management of men’s fertility either because of medical reasons, age, or other concerns.

During the last 60 years, numerous techniques have been developed and perfected in order to obtain good fertility through the application of frozen seminal doses. Despite similarities in most sperm cryopreservation protocols, differences exist between and within species due to breed or species characteristics and individual response of sperm to cryopreservation protocols. Although great advances in freezing semen and its utilization have been successful, the fertility of frozen thawed spermatozoa on different species is still low. Therefore, further knowledge and investigation are needed to improve the quality of frozen–thawed semen in various species.

In order to supply this demand, a Special Issue will be focused on exploring different approaches to provide revealing information as regards sperm processes for cryopreservation and their usefulness for further application in domestic, wild, and endangered animals.

Dr. Maria-Jesus Palomo Peiró
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

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. Animals is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2400 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • semen cryopreservation
  • spermatozoa
  • quality assessment
  • freeze-thawing
  • resilience
  • assisted reproduction technologies
  • fertility

Published Papers (6 papers)

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Research

13 pages, 2342 KiB  
Article
The Cryopreserved Sperm Traits of Various Ram Breeds: Towards Biodiversity Conservation
by Jakub Vozaf, Andrea Svoradová, Andrej Baláži, Jaromír Vašíček, Lucia Olexiková, Linda Dujíčková, Alexander V. Makarevich, Rastislav Jurčík, Hana Ďúranová and Peter Chrenek
Animals 2022, 12(10), 1311; https://0-doi-org.brum.beds.ac.uk/10.3390/ani12101311 - 20 May 2022
Cited by 8 | Viewed by 1780
Abstract
The aim of our research was to compare three Slovak sheep breeds in the quality parameters of cryopreserved sperm. The ejaculates of Slovak Dairy (SD), Native Wallachian (NW), and Improved Wallachian (IW) sheep rams (n = 12) were collected by electro-ejaculation. Heterospermic samples [...] Read more.
The aim of our research was to compare three Slovak sheep breeds in the quality parameters of cryopreserved sperm. The ejaculates of Slovak Dairy (SD), Native Wallachian (NW), and Improved Wallachian (IW) sheep rams (n = 12) were collected by electro-ejaculation. Heterospermic samples were created from suitable ejaculates, separately for each breed (at least 90% of total and 80% of progressive motility). Samples were equilibrated in a Triladyl® diluent and frozen by automated freezing. Sperm samples were subjected to the motility, morphology, (CASA), viability and apoptosis (DRAQ7/Yo-Pro-1), fertilizing capability (penetration/fertilization test (P/F) in vitro) and acrosomal status (transmission electron microscopy) assays before freezing and after thawing. It was found that there were no significant differences (p < 0.05) between the evaluated breeds in motility, viability, apoptosis, morphological properties, and fertilizing ability of cryopreserved sperm. Significant differences occurred in acrosomal status. Our results demonstrate that the use of the selected cryopreservation protocol is suitable for at least three different sheep breeds, which can greatly benefit the biodiversity protection and simplifies the creation of an animal genetic resources gene bank. Full article
(This article belongs to the Special Issue Further Approaches on Sperm Cryopreservation)
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16 pages, 2316 KiB  
Article
Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and Pentaisomaltose
by Ondrej Simonik, Filipa Bubenickova, Lucie Tumova, Michaela Frolikova, Vishma Pratap Sur, Jan Beran, Katerina Havlikova, Lenka Hackerova, Daniela Spevakova, Katerina Komrskova and Pavla Postlerova
Animals 2022, 12(7), 868; https://0-doi-org.brum.beds.ac.uk/10.3390/ani12070868 - 30 Mar 2022
Cited by 7 | Viewed by 3597
Abstract
The long-term storage of boar sperm presents an ongoing challenge, and the modification of the cryoprotective compounds in semen extenders is crucial for improving cryopreservation’s success rate. The aim of our study was to reduce the percentage of glycerol in the extender by [...] Read more.
The long-term storage of boar sperm presents an ongoing challenge, and the modification of the cryoprotective compounds in semen extenders is crucial for improving cryopreservation’s success rate. The aim of our study was to reduce the percentage of glycerol in the extender by elimination or substitution with biocompatible, non-toxic polysaccharides. For boar semen extender improvement, we tested a novel modification with the polysaccharides dextran and pentaisomaltose in combination with unique in silico predictive modeling. We targeted the analysis of in vitro qualitative sperm parameters such as motility, viability, mitochondrial activity, acrosome integrity, and DNA integrity. Non-penetrating polysaccharide-based cryoprotective agents interact with sperm surface proteins such as spermadhesins, which are recognized as fertility markers of boar sperm quality. The in silico docking study showed a moderate binding affinity of dextran and pentaisomaltose toward one specific spermadhesin known as AWN, which is located in the sperm plasma membrane. Pentaisomaltose formed a hydrophobic pocket for the AWN protein, and the higher energy of this protein–ligand complex compared with dextran was calculated. In addition, the root mean square deviation (RMSD) analysis for the molecular dynamics (MD) of both polysaccharides and AWN simulation suggests their interaction was highly stable. The in silico results were supported by in vitro experiments. In the experimental groups where glycerol was partially or entirely substituted, the use of pentaisomaltose resulted in improved sperm mitochondrial activity and DNA integrity after thawing when compared with dextran. In this paper, we demonstrate that pentaisomaltose, previously used for cryopreservation in hematopoietic stem cells, represents a promising compound for the elimination or reduction of glycerol in extenders for boar semen cryopreservation. This novel approach, using in silico computer prediction and in vitro testing, represents a promising technique to help identify new cryoprotectants for use in animal breeding or genetic resource programs. Full article
(This article belongs to the Special Issue Further Approaches on Sperm Cryopreservation)
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10 pages, 7931 KiB  
Article
Cryopreservation of Giraffe Epidydimal Spermatozoa Using Different Extenders and Cryoprotectants
by Robert Hermes, Alexis Lecu, Romain Potier, Frank Goeritz, Jessica P. Rickard, Julia Bohner, Rudy Wedlarski, Jiri Hruby and Thomas B. Hildebrandt
Animals 2022, 12(7), 857; https://0-doi-org.brum.beds.ac.uk/10.3390/ani12070857 - 29 Mar 2022
Cited by 2 | Viewed by 2220
Abstract
Giraffe numbers have plummeted over the last 30 years by 30–40%. Thus, their conservation status has been raised from least concern to vulnerable. Efforts to manage in situ and ex situ populations are increasing. Assisted reproduction techniques (ART) such as sperm cryopreservation could [...] Read more.
Giraffe numbers have plummeted over the last 30 years by 30–40%. Thus, their conservation status has been raised from least concern to vulnerable. Efforts to manage in situ and ex situ populations are increasing. Assisted reproduction techniques (ART) such as sperm cryopreservation could help preserve the genetic diversity of giraffe subspecies and, when used for artificial inseminations, enhance genetic exchange between isolated populations. However, to date, the post-thaw motility of recovered sperm has been low and inconsistent. In this study, epididymal sperm collected from the testes of giraffes (n = 7) was frozen in three different extenders, namely, BotuCrio, Steridyl, and test egg yolk (TEY), each supplemented with one of two different cryoprotectants (5% glycerol or a mix of 1% glycerol and 4% methylformamide) and frozen over liquid nitrogen vapor. Across all three extenders, sperm showed significantly better post-thaw results when frozen with a mix of glycerol and methylformamide compared with glycerol alone. Sperm frozen with TEY and a mix of glycerol and methylformamide achieved superior post-thaw total and progressive sperm motility of 57 ± 3% and 45 ± 3%, respectively. These results show the benefit of using alternative cryoprotectants for freezing giraffe spermatozoa and could aid in the application of ARTs for giraffe subspecies or the closely related endangered Okapi. Full article
(This article belongs to the Special Issue Further Approaches on Sperm Cryopreservation)
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9 pages, 561 KiB  
Article
The Concentration of ProAKAP4 and Other Indicators of Cryopotential of Spermatozoa Cryopreserved in Extender with Holothuroidea Extract Addition
by Alicja Kowalczyk, Elżbieta Gałęska and Anna Bubel
Animals 2022, 12(4), 521; https://0-doi-org.brum.beds.ac.uk/10.3390/ani12040521 - 20 Feb 2022
Cited by 7 | Viewed by 1884
Abstract
The aim of this study was to determine the concentration of proAKAP4 and other indicators of cryopotential of spermatozoa cryopreserved in extender with Holothuroidea extract addition. Nine Holstein Friesian bulls, 3.5 years old, of known health status, were used for the study. The [...] Read more.
The aim of this study was to determine the concentration of proAKAP4 and other indicators of cryopotential of spermatozoa cryopreserved in extender with Holothuroidea extract addition. Nine Holstein Friesian bulls, 3.5 years old, of known health status, were used for the study. The animals were kept and fed equally. Semen was collected once a week using an artificial vagina. The commercially available Holothuroidea extract was used as a supplement to the commercial extender (0, 2, 4 and 6 µL/mL) before the freezing/thawing process. The viability, motility, motion parameters, and acrosome integrity of the sperm were analyzed with (test) or without (control) extract samples. Furthermore, the concentration of the proAKAP4 biomarker in frozen sperm was assessed. It was shown that the addition of 4 and 6 µL of the extract may have a positive effect on the quality parameters of the sperm after thawing. The results indicate that extender supplementation with the above extract modulates (increases) the concentration of proAKAP4 in sperm at all tested levels. Additionally, this indicator has become helpful in identifying sperm of poor biological quality. Moreover, it has been proven that the proAKAP4 biomarker can be successfully used to evaluate the effectiveness of the use of various extenders for semen cryopreservation. Full article
(This article belongs to the Special Issue Further Approaches on Sperm Cryopreservation)
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14 pages, 309 KiB  
Article
The Re-Addition of Seminal Plasma after Thawing Does Not Improve Buck Sperm Quality Parameters
by Uchechi Linda Ohaneje, Uchebuchi Ike Osuagwuh, Manuel Alvarez-Rodríguez, Iván Yánez-Ortiz, Abigail Tabarez and María Jesús Palomo
Animals 2021, 11(12), 3452; https://0-doi-org.brum.beds.ac.uk/10.3390/ani11123452 - 03 Dec 2021
Cited by 1 | Viewed by 1558
Abstract
In order to achieve a higher post-thaw buck sperm quality, an approach in the thawing protocol of cryopreserved sperm doses under in vitro capacitation conditions mimicking the in vivo female environment was studied. Therefore, functional and kinetic characteristics of buck thawed sperm from [...] Read more.
In order to achieve a higher post-thaw buck sperm quality, an approach in the thawing protocol of cryopreserved sperm doses under in vitro capacitation conditions mimicking the in vivo female environment was studied. Therefore, functional and kinetic characteristics of buck thawed sperm from males of different ages, the season of collection, and melatonin implanted males in the non-breeding season were assessed after 3 h of incubation in an in vitro fertilization (IVF) media with 20% of buck seminal plasma (SP). Previously, fresh ejaculates were collected via artificial vagina from eight males of the Cabra Blanca de Rasquera breed during two consecutive years in breeding and non-breeding periods. Prior to semen collection in non-breeding seasons, males were split into two groups: one group was implanted with melatonin, while the other was not. In each group, semen samples were pooled, centrifuged, and diluted in an extender containing 15% powdered egg yolk and 5% glycerol before freezing. After thawing, sperm were washed and incubated in three different media: (a) control media (modified phosphate-buffered saline (PBS), (b) IVF commercial media, and (c) IVF media + 20% SP. Sperm motility was evaluated by CASA, while plasma and acrosome membrane integrity, mitochondria activity, and DNA fragmentation were analysed by flow cytometer at 0 h and after 3 h incubation. A significant reduction in motility, mitochondrial activity, plasma, and acrosome membrane integrity were observed after incubation in the presence of SP, although similar to that observed in IVF media alone. DNA integrity was not affected under in vitro capacitation conditions, regardless of SP addition. In conclusion, the addition of SP failed to improve post-thaw buck sperm quality under in vitro conditions irrespective of male age, the season of collection, and melatonin implant. Full article
(This article belongs to the Special Issue Further Approaches on Sperm Cryopreservation)
14 pages, 2520 KiB  
Article
Association between Fatty Acid Composition, Cryotolerance and Fertility Competence of Progressively Motile Bovine Spermatozoa
by Tanya Kogan, Dana Grossman Dahan, Ronit Laor, Nurit Argov-Argaman, Yoel Zeron, Alisa Komsky-Elbaz, Dorit Kalo and Zvi Roth
Animals 2021, 11(10), 2948; https://0-doi-org.brum.beds.ac.uk/10.3390/ani11102948 - 13 Oct 2021
Cited by 7 | Viewed by 1956
Abstract
An association between progressive motility (PM) and spermatozoa fertility competence has been suggested. However, the mechanism that underlies PM is not clear enough. We examined physiological characteristics and fatty acid composition of fresh spermatozoa with high and low PM. Additional analysis of fatty [...] Read more.
An association between progressive motility (PM) and spermatozoa fertility competence has been suggested. However, the mechanism that underlies PM is not clear enough. We examined physiological characteristics and fatty acid composition of fresh spermatozoa with high and low PM. Additional analysis of fatty acid composition and structural characteristics was performed on spermatozoa samples with high and low progressively motile spermatozoa’s survival (PMSS), i.e., the ratio between the proportion of progressively motile spermatozoa after and before cryopreservation. Finally, a fertility field trial was conducted to examine the association between the number of PM spermatozoa within the insemination straw post thawing and conception rate. Analysis of fresh spermatozoa revealed a higher omega-6 to omega-3 ratio in ejaculates with low PM relative to those with high PM (p < 0.01). The proportion of polyunsaturated fatty acids was higher in low-PMSS fresh samples (p < 0.05) relative to their high-PMSS counterparts. Fresh samples with high-PMSS expressed a higher mitochondrial membrane potential (p < 0.05) and a higher proportion of viable cells that expressed reactive oxygen species (ROS; p < 0.05). Post-thawing evaluation revealed a reduced proportion of progressively motile sperm, with a prominent effect in samples with high PM relative to low PM, defined before freezing (p < 0.01). No differences in spermatozoa mitochondrial membrane potential or ROS level were found post-thawing. A fertility study revealed a positive correlation between the number of progressively motile spermatozoa within a standard insemination straw and conception rate (p < 0.05). Considering these, the bull PMSS is suggested to be taken into account at the time of straw preparation. Full article
(This article belongs to the Special Issue Further Approaches on Sperm Cryopreservation)
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