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Factors Affecting In Vitro Assessment of Sperm Quality
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Factors Affecting In Vitro Assessment of Sperm Quality

A special issue of Biology (ISSN 2079-7737). This special issue belongs to the section "Medical Biology".

Deadline for manuscript submissions: closed (31 October 2021) | Viewed by 26807

Special Issue Editors

Dr. Miguel A. Silvestre

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Guest Editor
Celular Biology, Functional Biology and Physical Antropology, University of Valencia, Valencia, Spain
Interests: spermatozoa; sperm assessment, cryopreservation; in vitro fertilization, artificial insemination
Dr. Jesús L. Yániz

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Guest Editor
Institute of Environmental Sciences (IUCA), University of Zaragoza, Huesca, Spain
Interests: animal reproduction; spermatology; artificial insemination; preservation of animal resources
Dr. Eva Mocé

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Guest Editor
Valencian Institute of Agricultural Research (IVIA), Center of Animal Technology (CITA), Segorbe, Spain
Interests: animal reproduction; andrology; cryobiology; preservation of animal resources
Dr. Carles Soler

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Guest Editor
Department of Cellular Biology, Functional Biology and Physical Anthropology, University of Valencia, Campus Burjassot, C/Dr Moliner 50, 46100 Burjassot, Spain
Interests: sperm motility and kinematics; sperm morphology and morphometrics; CASA technology applications; DNA fragmentation; maturation and stability
Prof. Dr. Eduardo R. S. Roldán

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Guest Editor
Department of Biodiversity and Evolutionary Biology, Museo Nacional de Ciencias Naturales (CSIC), Madrid, Spain
Interests: sperm biology; fertilization; reproductive biology; evolution; history and philosophy of biology
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The study of the different in vitro sperm parameters is essential to evaluate sperm quality, to predict the fertilizing potential of semen samples and to detect male infertility. These aspects are important in the development of Assisted Reproduction Technologies (IVF, ICSI) in human, the application of artificial insemination in livestock species, the conservation of animal biodiversity and in the study of factors with possible effect on male fertility. Despite the advances in recent years, the predictive ability of in vitro analysis on the fertility potential of semen remains still limited. The combination of different procedures of sperm evaluation, mainly flow cytometry and computer-assisted sperm analysis (CASA), as well as obtaining new knowledge about sperm parameters, could improve this predictive ability. Flow cytometry and CASA systems allow to objectively assess a high number of cells and parameters in a short period of time.  The fact that sperm is the most diverse animal cell in the animal kingdom implies that sperm quality analysis techniques often have to be adapted for each species. This special issue will focus on the most used and advanced in vitro methodologies to evaluate sperm quality for its application in the preservation of seminal doses and the prediction of semen and male fertility in different animal groups, such as mammals, birds, fishes or insects.

Dr. Miguel A. Silvestre
Dr. Jesús L. Yániz
Dr. Eva Mocé
Dr. Carles Soler 
Dr. Eduardo Roldán
Guest Editors

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. Biology 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 2700 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
  • In vitro assessment
  • Cytometry
  • Computer-Assisted Sperm Analysis
  • Male fertility
  • Artificial Insemination
  • sperm preservation
  • Assisted Reproductive Technologies
  • Andrology
  • Classification criteria

Published Papers (7 papers)

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Editorial

Jump to: Research, Review

3 pages, 226 KiB  
Editorial
The Importance of Studying Factors That Affect the In Vitro Evaluation of Semen Quality to Predict Potential Fertility in Males
by Miguel Angel Silvestre, Carles Soler, Eva Mocé, Eduardo R. S. Roldan and Jesús L. Yániz
Biology 2023, 12(2), 235; https://0-doi-org.brum.beds.ac.uk/10.3390/biology12020235 - 02 Feb 2023
Viewed by 1230
Abstract
The presence of sub-fertile or infertile males in farms or artificial insemination (AI) centres has a great impact on the reproductive and economic performance of the livestock industry [...] Full article
(This article belongs to the Special Issue Factors Affecting In Vitro Assessment of Sperm Quality)

Research

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17 pages, 1836 KiB  
Article
Impact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions
by Patricia Peris-Frau, Irene Sánchez-Ajofrín, Alicia Martín Maestro, Carolina Maside, Daniela Alejandra Medina-Chávez, Olga García-Álvarez, María del Rocío Fernández-Santos, Vidal Montoro, José Julián Garde, Manuel Ramón and Ana Josefa Soler
Biology 2021, 10(11), 1213; https://0-doi-org.brum.beds.ac.uk/10.3390/biology10111213 - 20 Nov 2021
Cited by 5 | Viewed by 1902
Abstract
The heterogeneous nature of ejaculates highlights the relevance of studying the behavior of different sperm subpopulations. Changes in sperm motility and the increase in tyrosine phosphorylation are key events that usually occur during capacitation and can be modified by the cryopreservation process. However, [...] Read more.
The heterogeneous nature of ejaculates highlights the relevance of studying the behavior of different sperm subpopulations. Changes in sperm motility and the increase in tyrosine phosphorylation are key events that usually occur during capacitation and can be modified by the cryopreservation process. However, the relationship between both events remains poorly defined throughout capacitation in the different sperm subpopulations. Fresh and frozen-thawed spermatozoa were incubated in capacitating (CAP) and non-capacitating (NC) media up to 240 min. Sperm kinematics, tyrosine phosphorylation and mitochondrial activity were measured by the CASA system and imaging flow cytometry. Four motile sperm subpopulations (SP) were identified in fresh and frozen-thawed ram semen after the cluster analysis. Incubation under CAP conditions over time led to greater changes in the percentage of spermatozoa included in each subpopulation compared to NC conditions, being different between fresh and frozen-thawed spermatozoa. The SP1, characterized by slow spermatozoa, progressively increased after 15 min in frozen-thawed samples incubated in both media but not in fresh ones. The SP4, characterized by fast and non-linear spermatozoa, showed a marked increase during CAP, but not under NC conditions, occurring more rapidly in frozen-thawed spermatozoa. This subpopulation (SP4) was also the only one positively and strongly correlated with mitochondrial activity and all phosphorylated sperm regions during capacitation, either in fresh or frozen-thawed samples. Our results indicated that in vitro capacitation induced significant changes in the distribution of motile sperm subpopulations, being affected by cryopreservation. Notwithstanding, the subpopulation which probably represents hyperactivated-like spermatozoa (SP4) also increased in frozen-thawed samples, occurring faster and simultaneously to the increment of mitochondrial activity and tyrosine phosphorylation of different sperm regions. Full article
(This article belongs to the Special Issue Factors Affecting In Vitro Assessment of Sperm Quality)
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17 pages, 1073 KiB  
Article
Characterization of Bottlenose Dolphin (Tursiops truncatus) Sperm Based on Morphometric Traits
by María del Carmen Fuentes-Albero, Silvia Abril Sánchez, José Luis Ros-Santaella, Eliana Pintus, Chiara Luongo, Sara Ruiz Díaz, Carlos Barros García, María Jesús Sánchez Calabuig, Daniel García Párraga and Francisco Alberto García Vázquez
Biology 2021, 10(5), 355; https://0-doi-org.brum.beds.ac.uk/10.3390/biology10050355 - 22 Apr 2021
Cited by 2 | Viewed by 3378
Abstract
Bottlenose dolphin (Tursiops truncatus) males follow many reproductive strategies to ensure their paternity. However, little is known about the sperm traits, including morphometric features, that contribute to their reproductive success. Our aim was to study dolphin sperm morphometry (a total of [...] Read more.
Bottlenose dolphin (Tursiops truncatus) males follow many reproductive strategies to ensure their paternity. However, little is known about the sperm traits, including morphometric features, that contribute to their reproductive success. Our aim was to study dolphin sperm morphometry (a total of 13 parameters) in two adult males to evaluate (i) presumptive sperm subpopulations, (ii) the correlation of sperm morphometry with testosterone levels and (iii) the effect of refrigerated storage on the sperm morphometry. Sperm populations were classified into four principal components (PCs) based on morphometry (>94% of cumulative variance). The PCs clustered into two different sperm subpopulations, which differed between males. Furthermore, the levels of serum testosterone were positively correlated with the length of the midpiece but negatively correlated with head width and the principal piece, flagellum and total sperm lengths. Most of the sperm morphometric parameters changed during the storage period (day 1 vs. day 7), but only the principal piece length was affected by the storage temperature (5 °C vs. 15 °C). This is the first study to identify dolphin sperm subpopulations based on morphometry and the influence of serum testosterone and refrigeration on sperm morphometry. Full article
(This article belongs to the Special Issue Factors Affecting In Vitro Assessment of Sperm Quality)
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13 pages, 797 KiB  
Article
Effect of Sperm Concentration and Storage Temperature on Goat Spermatozoa during Liquid Storage
by Sara Sadeghi, Raquel Del Gallego, Balma García-Colomer, Ernesto A. Gómez, Jesús L. Yániz, Jaime Gosálvez, Carmen López-Fernández and Miguel A. Silvestre
Biology 2020, 9(9), 300; https://0-doi-org.brum.beds.ac.uk/10.3390/biology9090300 - 19 Sep 2020
Cited by 15 | Viewed by 4279
Abstract
The use of cooled semen is relatively common in goats. There are a number of advantages of cooled semen doses, including easier handling of artificial insemination (AI) doses, transport, more AI doses per ejaculate, and higher fertility rates in comparison with frozen AI [...] Read more.
The use of cooled semen is relatively common in goats. There are a number of advantages of cooled semen doses, including easier handling of artificial insemination (AI) doses, transport, more AI doses per ejaculate, and higher fertility rates in comparison with frozen AI doses. However, cooled semen has a short shelf life. The objective of this study was to examine the effect of temperature and sperm concentration on the in vitro sperm quality during liquid storage for 48 h, including sperm motility and kinetics, response to oxidation, mitochondrial membrane potential (MMP) and DNA fragmentation in goats. Three experiments were performed. In the first, the effects of liquid preservation of semen at different temperatures (5 °C or 17 °C), durations (0, 24 and 48 h) and sperm concentrations (250 × 106 sperm/mL (1:2 dilution rate), 166.7 × 106 sperm/mL (1:3 dilution rate) or 50 × 106 sperm/mL (1:10 dilution rate)) on sperm motility and kinetics were studied. In the second experiment, the effect of temperature, sperm washing and concentration on sperm motility and DNA fragmentation was studied. Finally, the effect of sperm concentration and duration of storage at 5 °C on sperm motility, response to oxidative stress and MMP was examined. We found that refrigerated liquid storage of goat sperm impaired sperm quality, such as motility, MMP and response to oxidation, as storage time increased; however, sperm DNA fragmentation index was not significantly affected. Liquid storage at 5 °C preserved higher total motility than at 17 °C. Moreover, we observed that the reduction of sperm concentration below 500 × 106 sperm/mL did not seem to improve the quality of spermatozoa conserved in milk-based extender in the conditions tested. Full article
(This article belongs to the Special Issue Factors Affecting In Vitro Assessment of Sperm Quality)
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16 pages, 3333 KiB  
Article
Expanding the Limits of Computer-Assisted Sperm Analysis through the Development of Open Software
by Jesús Yániz, Carlos Alquézar-Baeta, Jorge Yagüe-Martínez, Jesús Alastruey-Benedé, Inmaculada Palacín, Sergii Boryshpolets, Vitaliy Kholodnyy, Hermes Gadêlha and Rosaura Pérez-Pe
Biology 2020, 9(8), 207; https://0-doi-org.brum.beds.ac.uk/10.3390/biology9080207 - 05 Aug 2020
Cited by 7 | Viewed by 4478
Abstract
Computer assisted sperm analysis (CASA) systems can reduce errors occurring in manual analysis. However, commercial CASA systems are frequently not applicable at the forefront of challenging research endeavors. The development of open source software may offer important solutions for researchers working in related [...] Read more.
Computer assisted sperm analysis (CASA) systems can reduce errors occurring in manual analysis. However, commercial CASA systems are frequently not applicable at the forefront of challenging research endeavors. The development of open source software may offer important solutions for researchers working in related areas. Here, we present an example of this, with the development of three new modules for the OpenCASA software (hosted at Github). The first is the Chemotactic Sperm Accumulation Module, a powerful tool for studying sperm chemotactic behavior, analyzing the sperm accumulation in the direct vicinity of the stimuli. This module was validated by comparing fish sperm accumulation, with or without the influence of an attractant. The analysis clearly indicated cell accumulation in the treatment group, while the distribution of sperm was random in the control group. The second is the Sperm Functionality Module, based on the ability to recognize five sperm subpopulations according to their fluorescence patterns associated with the plasma membrane and acrosomal status. The last module is the Sperm Concentration Module, which expands the utilities of OpenCASA. These last two modules were validated, using bull sperm, by comparing them with visual counting by an observer. A high level of correlation was achieved in almost all the data, and a good agreement between both methods was obtained. With these newly developed modules, OpenCASA is consolidated as a powerful free and open-source tool that allows different aspects of sperm quality to be evaluated, with many potential applications for researchers. Full article
(This article belongs to the Special Issue Factors Affecting In Vitro Assessment of Sperm Quality)
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16 pages, 516 KiB  
Article
Kinematic Sub-Populations in Bull Spermatozoa: A Comparison of Classical and Bayesian Approaches
by Luis Víquez, Vinicio Barquero, Carles Soler, Eduardo R.S. Roldan and Anthony Valverde
Biology 2020, 9(6), 138; https://0-doi-org.brum.beds.ac.uk/10.3390/biology9060138 - 26 Jun 2020
Cited by 22 | Viewed by 3975
Abstract
The ejaculate is heterogenous and sperm sub-populations with different kinematic patterns can be identified in various species. Nevertheless, although these sub-populations are statistically well defined, the statistical differences are not always relevant. The aim of the present study was to characterize kinematic sub-populations [...] Read more.
The ejaculate is heterogenous and sperm sub-populations with different kinematic patterns can be identified in various species. Nevertheless, although these sub-populations are statistically well defined, the statistical differences are not always relevant. The aim of the present study was to characterize kinematic sub-populations in sperm from two bovine species, and diluted with different commercial extenders, and to determine the statistical relevance of sub-populations through Bayesian analysis. Semen from 10 bulls was evaluated after thawing. An ISAS®v1 computer-assisted sperm analysis (CASA)-Mot system was employed with an image acquisition rate of 50 Hz and ISAS®D4C20 counting chambers. Sub-populations of motile spermatozoa were characterized using multivariate procedures such as principal components (PCs) analysis and clustering methods (k-means model). Four different sperm sub-populations were identified from three PCs that involved progressiveness, velocity, and cell undulatory movement. The proportions of the different sperm sub-populations varied with the extender used and in the two species. Despite a statistical difference (p < 0.05) between extenders, the Bayesian analysis confirmed that only one of them (Triladyl®) presented relevant differences in kinematic patterns when compared with Tris-EY and OptiXcell®. Extenders differed in the proportion of sperm cells in each of the kinematic sub-populations. Similar patterns were identified in Bos taurus and Bos indicus. Bayesian results indicate that sub-populations SP1, SP2, and SP3 were different for PC criteria and these differences were relevant. For velocity, linearity, and progressiveness, the SP4 did not show a relevant difference regarding the other sperm sub-populations. The classical approach of clustering or sperm subpopulation thus may not have a direct biological meaning. Therefore, the biological relevance of sperm sub-populations needs to be reevaluated. Full article
(This article belongs to the Special Issue Factors Affecting In Vitro Assessment of Sperm Quality)
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Review

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16 pages, 627 KiB  
Review
Sperm Quality Assessment in Honey Bee Drones
by Jesús L. Yániz, Miguel A. Silvestre and Pilar Santolaria
Biology 2020, 9(7), 174; https://0-doi-org.brum.beds.ac.uk/10.3390/biology9070174 - 18 Jul 2020
Cited by 23 | Viewed by 5496
Abstract
The quality of honey bee drone semen is relevant in different contexts, ranging from colony productivity to pathology, toxicology and biodiversity preservation. Despite its importance, considerably less knowledge is available on this subject for the honey bee when compared to other domestic animal [...] Read more.
The quality of honey bee drone semen is relevant in different contexts, ranging from colony productivity to pathology, toxicology and biodiversity preservation. Despite its importance, considerably less knowledge is available on this subject for the honey bee when compared to other domestic animal species. A proper assessment of sperm quality requires a multiple testing approach which discriminates between the different aspects of sperm integrity and functionality. Most studies on drone semen quality have only assessed a few parameters, such as sperm volume, sperm concentration and/or sperm plasma membrane integrity. Although more recent studies have focused on a broader variety of aspects of semen quality, some techniques currently used in vertebrates, such as computer-assisted sperm analysis (CASA) or multiparametric sperm quality testing, still remain to be developed in the honey bee. This may be attributed to the particular sperm morphology and physiology in this species, requiring the development of technologies specifically adapted to it. This article reviews the present knowledge of sperm quality in honey bee drones, highlighting its peculiarities and proposing future lines of research. Full article
(This article belongs to the Special Issue Factors Affecting In Vitro Assessment of Sperm Quality)
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