Sperm Biotechnology in Animals Reproduction

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

Deadline for manuscript submissions: 25 February 2025 | Viewed by 16380

Special Issue Editors


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Guest Editor
Faculty of Veterinary Sciences, Department of Animal Medicine and Surgery, University of Murcia, 30071 Murcia, Spain
Interests: animal reproduction; reproductive biotechnologies related to spermatozoa and embryos

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Guest Editor
Department of Agriculture, School of Agricultural Sciences, University of Western Macedonia, 53100 Florina, Greece
Interests: animals reproduction; artificial insemination; IVF; semen technology

Special Issue Information

Dear Colleagues,

The current advances of sperm technology, artificial insemination with liquid-/cryopreserved or sexed semen, depending on species, have been widely applied in practice and have facilitated, among others, the distribution of genetically strong animals worldwide. However, there are many points related to sperm biotechnological techniques that require further investigation. The improvement of sperm survival during liquid- or cryo-preservation and sex-sorting processes or the increase of the recovery rate of good quality sperm during sperm selection techniques is a requirement that is always related to high fertility rates (either in vivo or in vitro). In this field, research has focused on the extender’s composition, the cryopreservation protocols, and the discovery of agents or mechanism that could improve sperm fertilizing capacity. The molecular techniques have already given a new perspective to reproductive medicine that could further enhance our knowledge and give new insights into diagnosis by introducing new fertility markers. Modern diagnostic techniques for sperm quality and function evaluation could be applied for diagnosis in clinical practice or the prediction of animals’ fertility. New methods for sperm selection and handling in reproductive technologies could also increase the recovery rate of good quality sperm.

We are pleased to invite you to contribute to this Special Issue, which aims to publish original research articles or reviews on “Sperm biotechnology and its application”.

We invite researchers to contribute their recent findings, mainly focusing on, but not limited to, the following scientific topics:

  • Semen extenders;
  • Sperm technology;
  • Sperm sorting methods for assisted reproductive protocols;
  • New fertility markers for diagnosis;
  • Modern techniques for prediction of animal’s fertility;
  • Artificial insemination in domestic animals.

Dr. Inmaculada Parrilla Riera
Dr. Athina Basioura
Dr. Junwei Li
Guest Editors

Manuscript Submission Information

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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.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2100 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

  • spermatozoa
  • fertility
  • sperm fertilizing capacity
  • sperm sorting
  • assisted reproduction technologies
  • sperm biotechnological applications

Published Papers (7 papers)

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Research

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9 pages, 1122 KiB  
Article
Comparison of Nanoparticles and Single-Layer Centrifugation for Separation of Dead from Live Stallion Spermatozoa
by Christian Bisiau, Paula Moffett, James Graham and Patrick McCue
Vet. Sci. 2024, 11(7), 307; https://doi.org/10.3390/vetsci11070307 - 10 Jul 2024
Viewed by 361
Abstract
The goal of this study was to compare the efficacy of coated iron-core nanoparticles and single-layer centrifugation for separation of dead from live stallion spermatozoa. Our hypothesis was that nanoparticles would bind to dead sperm and allow for separation from live sperm using [...] Read more.
The goal of this study was to compare the efficacy of coated iron-core nanoparticles and single-layer centrifugation for separation of dead from live stallion spermatozoa. Our hypothesis was that nanoparticles would bind to dead sperm and allow for separation from live sperm using a magnet, resulting in a population of spermatozoa with a high percentage of total and progressive motility. Treatment Group 1 was an untreated control. Treatment Group 2 (nanoparticles, NP) utilized sperm incubated with nanoparticles followed by application of a magnet to remove dead sperm adhered to the coated nanoparticles. Treatment Group 3 (single-layer centrifugation, SLC) layered sperm above EquiPure™ followed by centrifugation. Semen samples were subsequently evaluated for sperm motility parameters, plasma membrane integrity, acrosome status, and morphology. The SLC technique yielded higher (p < 0.05) progressive motility (76 ± 9.2%) than the NP separation technique (59 ± 12.2%) or the untreated control (47.3 ± 5.1%). However, the total number of sperm recovered was higher (p < 0.05) in the NP technique (526.2 ± 96.6 × 106) than the SLC procedure (211.7 ± 70 × 106), yielding a higher total number of progressively motile sperm (317.6 ± 109 × 106) recovered using the NP technique than the SLC technique (157.8 ± 43.6 × 106). The percentage of live, acrosome intact sperm recovered was higher for SLC than NP. In summary, the SLC technique yielded a higher percentage of sperm motility, intact plasma membranes, and acrosome integrity, but yielded lower total sperm than with the nanoparticle separation technique. Full article
(This article belongs to the Special Issue Sperm Biotechnology in Animals Reproduction)
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15 pages, 2481 KiB  
Article
The Adaptation Time to the Extender as a Crucial Step for an Accurate Evaluation of Ram Sperm Quality during the Liquid Storage
by Marta Neila-Montero, Mercedes Alvarez, Marta F. Riesco, Cristina Soriano-Úbeda, Rafael Montes-Garrido, Cristina Palacin-Martinez, Paulino de Paz, Luis Anel and Luis Anel-Lopez
Vet. Sci. 2024, 11(3), 132; https://doi.org/10.3390/vetsci11030132 - 16 Mar 2024
Viewed by 1601
Abstract
Accurate assessment of ram sperm quality is crucial to optimizing assisted reproductive technologies in sheep. However, semen preservation can induce sperm due to osmotic, biochemical, and thermal stress. Stabilizing sperm with a suitable cooling rate and adaptation period to the extender could mitigate [...] Read more.
Accurate assessment of ram sperm quality is crucial to optimizing assisted reproductive technologies in sheep. However, semen preservation can induce sperm due to osmotic, biochemical, and thermal stress. Stabilizing sperm with a suitable cooling rate and adaptation period to the extender could mitigate these effects for a more reliable evaluation. This study aimed to determine: (1) the best time to assess ram sperm quality, and (2) the factor responsible for the altered state of ram sperm during the first hours of liquid storage. In Experiment 1, ejaculated sperm were diluted and assessed for sperm motility and functionality at four preservation times: 0, 3, 6, and 24 h as sperm damage control. Both sperm motility and functionality improved after 6 h. Experiment 2 investigated the factor responsible for sperm quality change by testing the interactions of seminal plasma and extender with sperm from epididymides independently and in combination. The evaluation of sperm was performed as in Experiment 1. Sperm in groups containing the extender showed altered motility at 0 and 24 h, and lower functionality at 0 h. Thus, we could assume that extender addition initially alters ram sperm, causing sublethal damage that is reversible after 3 to 6 h of semen preservation. In conclusion, ram sperm require an adaptation time of 3 to 6 h to the extender before an accurate quality assessment can be conducted. This has practical implications for reproduction centers, enabling better workflow organization and optimal expression of ram sperm attributes when cervical artificial insemination is routinely performed. Full article
(This article belongs to the Special Issue Sperm Biotechnology in Animals Reproduction)
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9 pages, 1645 KiB  
Communication
Investigating Visual Monitoring of the Scrotum as a Supplementary Tool for Boar Semen Quality Evaluation
by Vasiliki Stravogianni, Theodoros Samaras, Constantin M. Boscos, Athina Basioura, Ioannis Markakis and Ioannis A. Tsakmakidis
Vet. Sci. 2023, 10(1), 9; https://doi.org/10.3390/vetsci10010009 - 24 Dec 2022
Cited by 1 | Viewed by 2156
Abstract
Farm animals behavior research uses video cameras, mainly for visual observation and recording. The purpose of this feasibility study was to enrich the predictable methods of boar semen production capacity by correlating sperm variables with the scrotal contractions (SC) frequency and intensity. A [...] Read more.
Farm animals behavior research uses video cameras, mainly for visual observation and recording. The purpose of this feasibility study was to enrich the predictable methods of boar semen production capacity by correlating sperm variables with the scrotal contractions (SC) frequency and intensity. A video camera was used to record the reaction of the scrotum during ejaculation. The respective collected ejaculates were evaluated and semen parameters, such as viability, morphology, membranes functional integrity and kinematics, were determined. The camera recorded the scrotal contractions/relaxations and the video was handled by the Image Processing Toolbox of Matlab (Mathworks Inc., Natick, MA, USA). The SC intensity was verified as a percentage change in the scrotum size among the video frames of maximum contraction and relaxation. The archived data from the frames were analyzed statistically, using a linear mixed effects model that involved sperm assessed parameters. Correlations of the SC intensity with the average path velocity, VAP (R2 = 0.591, p = 0.043) and with the percentage of the cytoplasmic droplets (R2 = 0.509, p = 0.036) were noticed. Previous studies reported the positive correlation of VAP with the number of live-born piglets. In conclusion, video monitoring of the boar scrotal function during ejaculation is useful, but more research is needed to establish its appropriateness as a supplementary method for the prognosis of boar ability to produce high-quality semen. Full article
(This article belongs to the Special Issue Sperm Biotechnology in Animals Reproduction)
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16 pages, 1657 KiB  
Article
Proteomics Evaluation of Semen of Clinically Healthy Beagle-Breed Dogs
by Pagona G. Gouletsou, George Th. Tsangaris, Eleni I. Katsarou, Maria V. Bourganou, Mariana S. Barbagianni, Athina P. Venianaki, Efterpi Bouroutzika, Athanasios K. Anagnostopoulos, George C. Fthenakis and Angeliki I. Katsafadou
Vet. Sci. 2022, 9(12), 697; https://doi.org/10.3390/vetsci9120697 - 15 Dec 2022
Viewed by 2014
Abstract
The objectives of the present work were to evaluate the semen of dogs by means of proteomics methods and to compare with proteomics results of the blood of the animals, in order to increase available knowledge on the topic and present relevant reference [...] Read more.
The objectives of the present work were to evaluate the semen of dogs by means of proteomics methods and to compare with proteomics results of the blood of the animals, in order to increase available knowledge on the topic and present relevant reference values for semen samples. Semen samples were collected from five Beagle-breed dogs. Reproductive assessment of the animals by means of clinical, ultrasonographic and seminological examinations confirmed their reproductive health. The sperm-rich fraction and the prostatic fraction of semen were processed for proteomics evaluation. LC-MS/MS analysis was performed by means of a LTQ Orbitrap Elite system. The technology combines high separation capacity and strong qualitative ability of proteins in biological samples that require deep proteome coverage. Protein classification was performed based on their functional annotations using Gene Ontology (GO). In blood plasma, semen sperm-rich fraction, and semen prostatic fraction, 59, 42 and 43 proteins, respectively, were detected. Two proteins were identified simultaneously in plasma and the semen sperm-rich fraction, 11 proteins in plasma and the semen prostatic fraction, and three proteins in the semen sperm-rich and prostatic fractions. In semen samples, most proteins were related to cell organization and biogenesis, metabolic processes or transport of ions and molecules. Most proteins were located in the cell membrane, the cytosol or the nucleus. Finally, most proteins performed functions related to binding or enzyme regulation. There were no differences between the semen sperm-rich fraction and prostatic fractions in terms of the clustering of proteins. In conclusion, a baseline reference for proteins in the semen of Beagle-breed dogs is provided. These proteins are involved mostly in supporting spermatozoan maturation, survival and motility, enhancing the reproductive performance of male animals. There appears potential for the proteomics examination of semen to become a tool in semen evaluation. This analysis may potentially identify biomarkers for reproductive disorders. This can be particularly useful in stud animals, also given its advantage as a non-invasive method. Full article
(This article belongs to the Special Issue Sperm Biotechnology in Animals Reproduction)
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Review

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15 pages, 300 KiB  
Review
Freezing Stallion Semen—What Do We Need to Focus on for the Future?
by Ziyad Al-Kass and Jane M. Morrell
Vet. Sci. 2024, 11(2), 65; https://doi.org/10.3390/vetsci11020065 - 2 Feb 2024
Viewed by 3644
Abstract
Artificial insemination (AI) is used frequently in the breeding of sport horses, apart from Thoroughbreds. Most AIs are carried out with cooled semen rather than frozen semen because of the difficulties in identifying a protocol that is suitable for freezing most ejaculates and [...] Read more.
Artificial insemination (AI) is used frequently in the breeding of sport horses, apart from Thoroughbreds. Most AIs are carried out with cooled semen rather than frozen semen because of the difficulties in identifying a protocol that is suitable for freezing most ejaculates and the necessity to inseminate close to ovulation because of the short life of the thawed spermatozoa. More widespread use of frozen semen would improve biosecurity, allow greater choice of stallions, and offer more flexibility when managing deliveries of semen to the stud. It would even decrease the amount of antibiotics used in semen extenders, since the volume of frozen semen is smaller than when cooled semen is inseminated. However, there is considerable variability in the cryosurvival of spermatozoa from different stallions, leading to the classification of stallions as good or bad freezers. Improvements could be made at the level of stallion nutrition, the semen collection regimen, the extender, the removal of seminal plasma, and the cooling protocol, among others. Stallion sperm membranes are highly susceptible to lipid peroxidation, but research on antioxidants has failed to identify an additive that would benefit all stallions. In the future, biomarkers for sperm freezability could be used as an aid in identifying suitable ejaculates for cryopreservation. Full article
(This article belongs to the Special Issue Sperm Biotechnology in Animals Reproduction)
26 pages, 1694 KiB  
Review
Phospholipase C Zeta 1 (PLCZ1): The Function and Potential for Fertility Assessment and In Vitro Embryo Production in Cattle and Horses
by Raul A. Gonzalez-Castro and Elaine M. Carnevale
Vet. Sci. 2023, 10(12), 698; https://doi.org/10.3390/vetsci10120698 - 11 Dec 2023
Cited by 2 | Viewed by 2483
Abstract
Phospholipase C Zeta 1 (PLCZ1) is considered a major sperm-borne oocyte activation factor. After gamete fusion, PLCZ1 triggers calcium oscillations in the oocyte, resulting in oocyte activation. In assisted fertilization, oocyte activation failure is a major cause of low fertility. Most cases of [...] Read more.
Phospholipase C Zeta 1 (PLCZ1) is considered a major sperm-borne oocyte activation factor. After gamete fusion, PLCZ1 triggers calcium oscillations in the oocyte, resulting in oocyte activation. In assisted fertilization, oocyte activation failure is a major cause of low fertility. Most cases of oocyte activation failures in humans related to male infertility are associated with gene mutations and/or altered PLCZ1. Consequently, PLCZ1 evaluation could be an effective diagnostic marker and predictor of sperm fertilizing potential for in vivo and in vitro embryo production. The characterization of PLCZ1 has been principally investigated in men and mice, with less known about the PLCZ1 impact on assisted reproduction in other species, such as cattle and horses. In horses, sperm PLCZ1 varies among stallions, and sperm populations with high PLCZ1 are associated with cleavage after intracytoplasmic sperm injection (ICSI). In contrast, bull sperm is less able to initiate calcium oscillations and undergo nuclear remodeling, resulting in poor cleavage after ICSI. Advantageously, injections of PLCZ1 are able to rescue oocyte failure in mouse oocytes after ICSI, promoting full development and birth. However, further research is needed to optimize PLCZ1 diagnostic tests for consistent association with fertility and to determine whether PLCZ1 as an oocyte-activating treatment is a physiological, efficient, and safe method for improving assisted fertilization in cattle and horses. Full article
(This article belongs to the Special Issue Sperm Biotechnology in Animals Reproduction)
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17 pages, 395 KiB  
Review
The History and Prospects of Rabbit Sperm Sexing
by Patrícia Pinto-Pinho, Ana F. Ferreira, Rosário Pinto-Leite, Margarida Fardilha and Bruno Colaço
Vet. Sci. 2023, 10(8), 509; https://doi.org/10.3390/vetsci10080509 - 7 Aug 2023
Cited by 3 | Viewed by 2531
Abstract
Sperm sex selection is a longstanding challenge in the field of animal reproduction. The cuniculture industry, in particular producers of males or females for breeding purposes, would greatly benefit from the pre-selection of the offspring’s sex. This review article overviews the current and [...] Read more.
Sperm sex selection is a longstanding challenge in the field of animal reproduction. The cuniculture industry, in particular producers of males or females for breeding purposes, would greatly benefit from the pre-selection of the offspring’s sex. This review article overviews the current and future developments in rabbit sperm sexing technologies, as well as the implications of implementing these methodologies in cuniculture. The first attempts of sperm sexing were performed in rabbits; however, a both efficient and cost-effective methodology was not yet developed for this species. Those included sperm sexing according to differences in sperm density, surface electric charge, pH susceptibility, antisera reaction, and flow cytometry. Separation by flow cytometry has proven to be efficient in rabbits, yielding fractions with approximately 81% and 86% purity for X- and Y-sperm, respectively. However, it is not cost-effective for cuniculture and decreases sperm quality. The advantages, limitations, and practical considerations of each method are presented, highlighting their applicability and efficiency. Furthermore, herein we explore the potential of immunological-based techniques that overcome some of the limitations of earlier methods, as well as recent advancements in sperm sexing technologies in other animal models, which could be applied to rabbits. Finally, the challenges associated with the development and widespread implementation of rabbit sperm sexing technologies are addressed. By understanding the advantages and limitations of existing and emerging methods, researchers can direct their efforts towards the most promising directions, ultimately contributing to a more efficient, profitable, and sustainable cuniculture. Full article
(This article belongs to the Special Issue Sperm Biotechnology in Animals Reproduction)
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