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Animals
Special Issues
New Insights into Male Fertility and Sperm Preservation in Animals
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New Insights into Male Fertility and Sperm Preservation in Animals

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

Deadline for manuscript submissions: 30 April 2026 | Viewed by 3173

Special Issue Editor

Prof. Dr. Alexandre Rodrigues Silva

E-Mail Website
Guest Editor
Laboratory of Animal Germplasm Conservation, Department of Animal Sciences, Federal University of Semiarid Region—UFERSA, Mossoró 59625-900, RN, Brazil
Interests: wildlife reproductive physiology and biotechnologies; assisted reproductive technology; wildlife reproduction
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The development of methods to preserve male fertility and sperm in animals is crucial to the conservation of valuable genotypes and is essential for assisted reproduction programs, facilitating artificial insemination and embryo transfer, thereby increasing the efficiency of controlled breeding efforts. For domestic animals, studies in this area can contribute to the continuous improvement of breeds, increasing the productivity and sustainability of breeding systems. Furthermore, knowledge on this subject is highly applicable to wildlife, thus contributing to the preservation of biodiversity. Over several years, methodologies have focused mainly on improving sperm conservation protocols, whether from samples from ejaculates or obtained through recovery of the epididymis. More recently, it has been possible to observe a great advent in the development of methods of cryopreservation and culture of testicular tissues and cells, which have enabled the in vitro resumption of spermatogenesis, thus expanding the possibilities of strategies for preserving male fertility. In this sense, this issue aims to highlight new insights into the preservation of male fertility, showing not only advances in sperm preservation, but also highlighting the possibilities related to the conservation of gonadal tissues, both in domestic and wild animals.

Prof. Dr. Alexandre Rodrigues Silva
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
  • epidydimal sperm
  • testicular tissue
  • spermatogonia
  • biobank

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Published Papers (4 papers)

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Research

13 pages, 5411 KiB  
Article
Solid Surface Vitrification Is Better than Slow Freezing for the Long-Term Preservation of Testicular Fragments from Prepubertal Collared Peccaries (Pecari tajacu Linnaeus, 1758)
by Andréia M. Silva, Ana G. Pereira, Gabriel S. C. Bezerra, Yuri G. Matos, Luana G. P. Bezerra, Alexsandra F. Pereira, Moacir F. Oliveira, Pierre Comizzoli and Alexandre R. Silva
Animals 2025, 15(10), 1488; https://doi.org/10.3390/ani15101488 - 20 May 2025
Viewed by 300
Abstract
The cryopreservation of male gonadal tissue is critical to conserve genetic material and use it later via assisted reproduction. This study aimed to evaluate cryopreservation methods (slow freezing, SF; solid surface vitrification, SSV) as well as the optimal concentrations of intracellular cryoprotectants during [...] Read more.
The cryopreservation of male gonadal tissue is critical to conserve genetic material and use it later via assisted reproduction. This study aimed to evaluate cryopreservation methods (slow freezing, SF; solid surface vitrification, SSV) as well as the optimal concentrations of intracellular cryoprotectants during the SSV of testicular tissue from prepubertal collared peccaries. Five pairs of testes were dissected on different days into small fragments (3 mm3) and allocated to a non-cryopreserved, a control group or one of three treatment groups: SF; SSV 3 M (1.5 M dimethyl sulfoxide [DMSO] plus 1.5 M ethylene glycol [EG]); or SSV 6 M (3 M DMSO plus 3 M EG). After one week of storage in liquid nitrogen, tissue samples were warmed and evaluated in terms of histology, viability, proliferative capacity potential, and DNA integrity. The scores for histological integrity and cellular damage for SF (2.08 ± 0.05 and 2.33 ± 0.07, respectively) were similar to the results found in SSV 6 M (1.93 ± 0.04 and 2.30 ± 0.07; p > 0.05). However, these scores were better when compared to SSV 3 M (1.87 ± 0.05 and 2.08 ± 0.06; p < 0.05). The percentage of cellular viability was around 57% after all preservation treatments (p > 0.05), which was lower than in the control group (88.8 ± 1.9%; p < 0.05). The SSV 6 M treatment was better than the other treatments regarding the proliferative capacity potential of spermatogonia cells (3.52 ± 0.03) (p < 0.05), although it was lower than in the control group (4.00 ± 0.12) (p < 0.05). Additionally, SSV 6 M led to the same DNA integrity (97.0 ± 0.7%) as in the control group (99.4 ± 0.3%). These collective findings suggest that the combination of SSV with 6 M cryoprotectants is the most efficient for the cryopreservation of testes from prepubertal collared peccaries. Full article
(This article belongs to the Special Issue New Insights into Male Fertility and Sperm Preservation in Animals)
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9 pages, 1033 KiB  
Article
Effect of Tunicamycin on Viability, Motility, Reactive Oxygen Species, Nitric Oxide, and Lipid Peroxidation in Boar Sperm
by Seunghyung Lee
Animals 2025, 15(10), 1422; https://doi.org/10.3390/ani15101422 - 14 May 2025
Viewed by 303
Abstract
Tunicamycin induces endoplasmic reticulum stress in mammalian cells. Our study aimed to investigate the effect of tunicamycin on the motility and viability of sperm, reactive oxygen species, nitric oxide, and lipid peroxidation in boar sperm. We treated 1.0, 2.0, 5.0, and 10 μM [...] Read more.
Tunicamycin induces endoplasmic reticulum stress in mammalian cells. Our study aimed to investigate the effect of tunicamycin on the motility and viability of sperm, reactive oxygen species, nitric oxide, and lipid peroxidation in boar sperm. We treated 1.0, 2.0, 5.0, and 10 μM of tunicamycin in boar semen, and experimental treatments were performed. The viability (55.44%, 53.20, and 40.00%, p < 0.05) and motility (73.28%, 71.48%, and 54.48%, p < 0.05) of sperm at 2.0, 5.0, and 10.0 μM were decreased by tunicamycin, and the levels of reactive oxygen species and lipid peroxidation in tunicamycin-treated boar semen were increased (p < 0.05). However, the nitric oxide level was not changed by tunicamycin. Based on the results, we indicated that tunicamycin induces cell death by increasing oxidative stress in boar sperm, which may be the cause of decreased sperm viability and motility. Thus, we suggest that tunicamycin may induce cell death due to oxidative stress by reactive oxygen species and lipid peroxidation. Full article
(This article belongs to the Special Issue New Insights into Male Fertility and Sperm Preservation in Animals)
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14 pages, 4409 KiB  
Article
Letrozole and Crocin: Protecting Leydig Cells and Modulating Androgen Receptor and CYP19 Gene Expression in Busulfan-Induced Azoospermia
by Shahrzad Nokhbeh Zaeem, Mitra Heydari Nasrabadi, Masoud Salehipour and Somayeh Ehtesham
Animals 2025, 15(5), 697; https://doi.org/10.3390/ani15050697 - 27 Feb 2025
Viewed by 468
Abstract
This study aimed to investigate the impact of letrozole and crocin on Leydig cells on busulfan-induced azoospermia using a rat model. A sample population consisting of thirty male rats was randomly assigned to five groups: (1) the control group, (2) azoospermia group, (3) [...] Read more.
This study aimed to investigate the impact of letrozole and crocin on Leydig cells on busulfan-induced azoospermia using a rat model. A sample population consisting of thirty male rats was randomly assigned to five groups: (1) the control group, (2) azoospermia group, (3) azoospermia group treated with letrozole, (4) azoospermia group treated with crocin, and (5) azoospermia group treated with both letrozole and crocin. The treatment period with letrozole and crocin lasted for four weeks following busulfan administration. Subsequently, comprehensive analyses, including histopathological, molecular, and hormonal assessments, were performed, followed by immunohistochemical staining. This study found that the control group exhibited normal Leydig cell morphology, while the azoospermia group showed reduced Leydig cells and tissue disruptions. Letrozole and crocin treatments were associated with increased testicular fibrosis in the AZO and AZO + Cro groups, while their combination notably reduced fibrosis to approximately 15%. Furthermore, the combination treatment enhanced antioxidant enzyme activity and upregulated androgen receptor expression. Although a number of improvements were noted in sperm motility, they were not statistically significant. Further research is required to clarify the therapeutic implications of these findings in azoospermia treatment. Full article
(This article belongs to the Special Issue New Insights into Male Fertility and Sperm Preservation in Animals)
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15 pages, 1264 KiB  
Article
Effects of Supplementation of Different Antioxidants to Cryopreservation Extender on the Post-Thaw Quality of Rooster Semen—A Meta-Analysis
by Esther Díaz Ruiz, Francisco Javier Navas González, José Manuel León Jurado, Ander Arando Arbulu, Juan Vicente Delgado Bermejo and Antonio González Ariza
Animals 2024, 14(20), 2936; https://doi.org/10.3390/ani14202936 - 11 Oct 2024
Cited by 1 | Viewed by 1492
Abstract
The standardization of the semen cryopreservation technique could be an effective tool in poultry for the conservation of genetic resources. During this process, the production of reactive oxygen species increases, leading to oxidative stress that causes damage to the spermatozoa. To reduce this [...] Read more.
The standardization of the semen cryopreservation technique could be an effective tool in poultry for the conservation of genetic resources. During this process, the production of reactive oxygen species increases, leading to oxidative stress that causes damage to the spermatozoa. To reduce this effect, the addition of exogenous antioxidants in the cryopreservation diluent has been reported to be effective. Multiple antioxidants such as catalase, vitamin E, cysteamine, ergothioneine, and serine have been studied in roosters. Therefore, the present investigation aims to perform a meta-analysis to determine if the use of the aforementioned antioxidants added to the cryopreservation extender produces an improvement in semen quality parameters in roosters after thawing. After collecting the data, a discriminant canonical analysis was performed to determine which of the selected semen quality traits provided the most information, with hypo-osmotic swelling test (HOST), viability, and total motility variables showing the highest discriminatory power. However, according to the descriptive statistics, catalase and serine are the antioxidants that improve a greater number of seminal quality parameters, and since catalase gives the most favorable results for most of the discriminating variables, it could be the antioxidant of choice. Full article
(This article belongs to the Special Issue New Insights into Male Fertility and Sperm Preservation in Animals)
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