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The Era of Assisted Reproductive Technologies Tailored to the Specific...
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The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports

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

Deadline for manuscript submissions: closed (1 November 2020) | Viewed by 51834

Printed Edition Available!
A printed edition of this Special Issue is available here.

Special Issue Editor

Dr. David Martín Hidalgo

E-Mail Website1 Website2
Guest Editor
Department of Biochemistry, Molecular Biology and Genetics, Universidad de Extremadura, 10003 Cáceres, Spain
Interests: sperm; reproduction; fertility; Assisted Reproductive Technologies; cryopreservation; Artificial Insemination; ICSI; sex sorting; flow cytometry

Special Issue Information

Dear Colleagues,

Significant development of Assisted Reproductive Technologies (ART) has occurred in recent years. Prior to the first description of spermatozoa by Leeuwenhoek in 1677, Arabic tales described a primitive form of ART involving the transfer of semen from one mare into the vagina of another.

Nowadays, ART has a pivotal role not only in achieving fertilization in subfertile animals, but it is also involved in the management of the herd, the decrease of disease spread, and even allows the offspring sex selection. Nonetheless, there are differences between species or even intraspecies that have led researchers worldwide to focus on those differences in order to bypass these specific difficulties.

This Special Issue of Animals is open to original and review manuscripts that cover current issues in ART from biological and molecular points of view. The goal is to recognize the advances in ART in recent years as well as to describe the current problems and solutions continuing to the future of this astounding branch of the biology of reproduction. It will provide a balanced overview of emerging techniques and technologies that can be used to preserve, improve, salvage, or even create fertility for domestic animals with high economic weight in our society.

Dr. David Martín Hidalgo
Guest Editor

Manuscript Submission Information

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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

  • sperm
  • reproduction
  • fertility
  • assisted reproductive technologies
  • cryopreservation
  • artificial insemination
  • ICSI
  • sex sorting
  • flow cytometry
  • embryo
  • oocyte

Published Papers (15 papers)

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Research

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19 pages, 1914 KiB  
Article
The Effects of Red Light on Mammalian Sperm Rely upon the Color of the Straw and the Medium Used
by Jaime Catalán, Iván Yánez-Ortiz, Sabrina Gacem, Marion Papas, Sergi Bonet, Joan E. Rodríguez-Gil, Marc Yeste and Jordi Miró
Animals 2021, 11(1), 122; https://doi.org/10.3390/ani11010122 - 8 Jan 2021
Cited by 4 | Viewed by 2917
Abstract
Previous research has determined that irradiation of mammalian sperm with red light increases motility, mitochondrial activity, and fertilization capacity. In spite of this, no study has considered the potential influence of the color of the straw and the extender used. Therefore, this study [...] Read more.
Previous research has determined that irradiation of mammalian sperm with red light increases motility, mitochondrial activity, and fertilization capacity. In spite of this, no study has considered the potential influence of the color of the straw and the extender used. Therefore, this study tests the hypothesis that the response of mammalian sperm to red light is influenced by the color of the straw and the turbidity/composition of the extender. Using the horse as a model, 13 ejaculates from 13 stallions were split into two equal fractions, diluted with Kenney or Equiplus extender, and stored at 4 °C for 24 h. Thereafter, each diluted fraction was split into five equal aliquots and subsequently packed into 0.5-mL straws of red, blue, yellow, white, or transparent color. Straws were either nonirradiated (control) or irradiated with a light–dark–light pattern of 3–3–3 (i.e., light: 3 min, dark: 3 min; light: 3 min) prior to evaluating sperm motility, acrosome and plasma membrane integrity, mitochondrial membrane potential, and intracellular ROS and calcium levels. Our results showed that irradiation increased some motion variables, mitochondrial membrane potential, and intracellular ROS without affecting the integrities of the plasma membrane and acrosome. Remarkably, the extent of those changes varied with the color of the straw and the extender used; the effects of irradiation were more apparent when sperm were diluted with Equiplus extender and packed into red-colored straws or when samples were diluted with Kenney extender and packed into transparent straws. As the increase in sperm motility and intracellular ROS levels was parallel to that of mitochondrial activity, we suggest that the impact of red light on sperm function relies upon the specific rates of energy provided to the mitochondria, which, in turn, vary with the color of the straw and the turbidity/composition of the extender. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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18 pages, 2652 KiB  
Article
Effects of In Vitro Interactions of Oviduct Epithelial Cells with Frozen–Thawed Stallion Spermatozoa on Their Motility, Viability and Capacitation Status
by Brenda Florencia Gimeno, María Victoria Bariani, Lucía Laiz-Quiroga, Eduardo Martínez-León, Micaela Von-Meyeren, Osvaldo Rey, Adrián Ángel Mutto and Claudia Elena Osycka-Salut
Animals 2021, 11(1), 74; https://doi.org/10.3390/ani11010074 - 3 Jan 2021
Cited by 4 | Viewed by 2758
Abstract
Cryopreservation by negatively affecting sperm quality decreases the efficiency of assisted reproduction techniques (ARTs). Thus, we first evaluated sperm motility at different conditions for the manipulation of equine cryopreserved spermatozoa. Higher motility was observed when spermatozoa were incubated for 30 min at 30 [...] Read more.
Cryopreservation by negatively affecting sperm quality decreases the efficiency of assisted reproduction techniques (ARTs). Thus, we first evaluated sperm motility at different conditions for the manipulation of equine cryopreserved spermatozoa. Higher motility was observed when spermatozoa were incubated for 30 min at 30 × 106/mL compared to lower concentrations (p < 0.05) and when a short centrifugation at 200× g was performed (p < 0.05). Moreover, because sperm suitable for oocyte fertilization is released from oviduct epithelial cells (OECs), in response to the capacitation process, we established an in vitro OEC culture model to select a sperm population with potential fertilizing capacity in this species. We demonstrated E-cadherin and cytokeratin expression in cultures of OECs obtained. When sperm–OEC cocultures were performed, the attached spermatozoa were motile and presented an intact acrosome, suggesting a selection by the oviductal model. When co-cultures were incubated in capacitating conditions a greater number of alive (p < 0.05), capacitated (p < 0.05), with progressive motility (p < 0.05) and with the intact acrosome sperm population was observed (p < 0.05) suggesting that the sperm population released from OECs in vitro presents potential fertilizing capacity. Improvements in handling and selection of cryopreserved sperm would improve efficiencies in ARTs allowing the use of a population of higher-quality sperm. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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18 pages, 979 KiB  
Article
Selection of Boar Sperm by Reproductive Biofluids as Chemoattractants
by Luis Alberto Vieira, Alessia Diana, Cristina Soriano-Úbeda and Carmen Matás
Animals 2021, 11(1), 53; https://doi.org/10.3390/ani11010053 - 30 Dec 2020
Cited by 5 | Viewed by 2442
Abstract
Chemotaxis is a spermatozoa guidance mechanism demonstrated in vitro in several mammalian species including porcine. This work focused on follicular fluid (FF), periovulatory oviductal fluid (pOF), the medium surrounding oocytes during in vitro maturation (conditioned medium; CM), progesterone (P4), and the combination of [...] Read more.
Chemotaxis is a spermatozoa guidance mechanism demonstrated in vitro in several mammalian species including porcine. This work focused on follicular fluid (FF), periovulatory oviductal fluid (pOF), the medium surrounding oocytes during in vitro maturation (conditioned medium; CM), progesterone (P4), and the combination of those biofluids (Σ) as chemotactic agents and modulators of spermatozoa fertility in vitro. A chemotaxis chamber was designed consisting of two independent wells, A and B, connected by a tube. The spermatozoa are deposited in well A, and the chemoattractants in well B. The concentrations of biofluids that attracted a higher proportion of spermatozoa to well B were 0.25% FF, 0.25% OF, 0.06% CM, 10 pM P4 and 0.25% of a combination of biofluids (Σ2), which attracted between 3.3 and 12.3% of spermatozoa (p < 0.05). The motility of spermatozoa recovered in well B was determined and the chemotactic potential when the sperm calcium channel CatSper was inhibited, which significantly reduced the % of spermatozoa attracted (p < 0.05). Regarding the in vitro fertility, the spermatozoa attracted by FF produced higher rates of penetration of oocytes and development of expanded blastocysts. In conclusion, porcine reproductive biofluids show an in vitro chemotactic effect on spermatozoa and modulate their fertilizing potential. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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15 pages, 2126 KiB  
Article
Cellular and Molecular Events that Occur in the Oocyte during Prolonged Ovarian Storage in Sheep
by Alicia Martín-Maestro, Irene Sánchez-Ajofrín, Carolina Maside, Patricia Peris-Frau, Daniela-Alejandra Medina-Chávez, Beatriz Cardoso, José Carlos Navarro, María Rocío Fernández-Santos, José Julián Garde and Ana Josefa Soler
Animals 2020, 10(12), 2414; https://doi.org/10.3390/ani10122414 - 17 Dec 2020
Cited by 7 | Viewed by 2745
Abstract
For the past two decades, there has been a growing interest in the application of in vitro embryo production (IVP) in small ruminants such as sheep. To improve efficiency, a large number abattoir-derived ovaries must be used, and long distances from the laboratory [...] Read more.
For the past two decades, there has been a growing interest in the application of in vitro embryo production (IVP) in small ruminants such as sheep. To improve efficiency, a large number abattoir-derived ovaries must be used, and long distances from the laboratory are usually inevitable when adult animals are used. In that scenario, prolonged sheep ovary transportation may negatively affect oocyte developmental competence. Here, we evaluated the effect of ovary storage time (3, 5, 7, 9, 11 and 13 h) and the medium in which they were transported (TCM199 and saline solution) on oocyte quality. Thus, live/dead status, early apoptosis, DNA fragmentation, reduced glutathione (GSH) and reactive oxygen species (ROS) content, caspase-3 activity, mitochondrial membrane potential and distribution, and relative abundance of mRNA transcript levels were assessed in oocytes. After in vitro maturation (IVM), cumulus cell viability and quality, meiotic and fertilization competence, embryo rates and blastocyst quality were also evaluated. The results revealed that, after 7 h of storage, oocyte quality and developmental potential were significantly impaired since higher rates of dead oocytes and DNA fragmentation and lower rates of viable, matured and fertilized oocytes were observed. The percentage of cleavage, blastocyst rates and cumulus cell parameters (viability, active mitochondria and GSH/ROS ratio) were also decreased. Moreover, the preservation of ovaries in medium TCM199 had a detrimental effect on cumulus cells and oocyte competence. In conclusion, ovary transport times up to 5 h in saline solution are the most adequate storage conditions to maintain oocyte quality as well as developmental capacity in sheep. A strategy to rescue the poor developmental potential of stored oocytes will be necessary for successful production of high-quality embryos when longer ovarian preservation times are necessary. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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17 pages, 2922 KiB  
Article
Seminal Plasma Triggers the Differential Expression of the Glucocorticoid Receptor (NR3C1/GR) in the Rabbit Reproductive Tract
by Mateo Ruiz-Conca, Jaume Gardela, Amaia Jauregi-Miguel, Cristina A. Martinez, Heriberto Rodríguez-Martinez, Manel López-Béjar and Manuel Alvarez-Rodriguez
Animals 2020, 10(11), 2158; https://doi.org/10.3390/ani10112158 - 19 Nov 2020
Cited by 3 | Viewed by 2172
Abstract
Rabbits are interesting as research animal models for reproduction, due to their condition of species of induced ovulation, with the release of endogenous gonadotropin-releasing hormone (GnRH) due to coitus. Glucocorticoid (GC) signaling, crucial for physiological homeostasis, is mediated through a yet unclear mechanism, [...] Read more.
Rabbits are interesting as research animal models for reproduction, due to their condition of species of induced ovulation, with the release of endogenous gonadotropin-releasing hormone (GnRH) due to coitus. Glucocorticoid (GC) signaling, crucial for physiological homeostasis, is mediated through a yet unclear mechanism, by the GC receptor (NR3C1/GR). After mating, the female reproductive tract undergoes dynamic modifications, triggered by gene transcription, a pre-amble for fertilization and pregnancy. This study tested the hypothesis that when ovulation is induced, the expression of NR3C1 is influenced by sperm-free seminal plasma (SP), similarly to after mating (whole semen), along the different segments of the internal reproductive tract of female rabbits. Semen (mating) was compared to vaginal infusion of sperm-free SP (Experiment 1), and changes over time were also evaluated, i.e., 10, 24, 36, 68, and 72 h post-mating, corresponding to specific stages, i.e., ovulation, fertilization, and the interval of early embryo development up to the morula stage (Experiment 2). All does were treated with GnRH to induce ovulation. Samples were retrieved from seven segments of the reproductive tract (from the cervix to infundibulum), at 20 h post-mating or sperm-free SP infusion (Experiment 1) or at 10, 24, 36, 68, and 72 h post-mating (Experiment 2). Gene expression of NR3C1 was analyzed by qPCR. Results showed an increase in NR3C1 expression in the infundibulum compared to the other anatomical regions in the absence of spermatozoa when sperm-free SP infusion was performed (Experiment 1). Moreover, during the embryo transport through the oviduct, the distal isthmus was time-course upregulated, especially at 72 h, when morulae are retained in this anatomical region, while it was downregulated in the distal uterus at 68 h (Experiment 2). The overall results suggest that NR3C1, the GC receptor gene, assessed in the reproductive tract of does for the first time, shows differential expression changes during the interval of oviductal and uterine embryo transport that may imply a relevant role of the GC action, not only close to the site of ovulation and fertilization, but also in the endometrium. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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17 pages, 4023 KiB  
Article
Perfluorooctane Sulfonate (PFOS) and Perfluorohexane Sulfonate (PFHxS) Alters Protein Phosphorylation, Increase ROS Levels and DNA Fragmentation during In Vitro Capacitation of Boar Spermatozoa
by Iván Oseguera-López, Serafín Pérez-Cerezales, Paola Berenice Ortiz-Sánchez, Oscar Mondragon-Payne, Raúl Sánchez-Sánchez, Irma Jiménez-Morales, Reyna Fierro and Humberto González-Márquez
Animals 2020, 10(10), 1934; https://doi.org/10.3390/ani10101934 - 21 Oct 2020
Cited by 14 | Viewed by 3000
Abstract
Perfluorooctane sulfonate (PFOS) and perfluorohexane sulfonate (PFHxS) are toxic and bioaccumulative, included in the Stockholm Convention’s list as persistent organic pollutants. Due to their toxicity, worldwide distribution, and lack of information in spermatozoa physiology during pre-fertilization processes, the present study seeks to analyze [...] Read more.
Perfluorooctane sulfonate (PFOS) and perfluorohexane sulfonate (PFHxS) are toxic and bioaccumulative, included in the Stockholm Convention’s list as persistent organic pollutants. Due to their toxicity, worldwide distribution, and lack of information in spermatozoa physiology during pre-fertilization processes, the present study seeks to analyze the toxic effects and possible alterations caused by the presence of these compounds in boar sperm during the in vitro capacitation. The spermatozoa capacitation was performed in supplemented TALP-Hepes media and mean lethal concentration values of 460.55 μM for PFOS, and 1930.60 μM for PFHxS were obtained. Results by chlortetracycline staining showed that intracellular Ca2+ patterns bound to membrane proteins were scarcely affected by PFOS. The spontaneous acrosome reaction determined by FITC-PNA was significantly reduced by PFOS and slightly increased by PFHxS. Both toxic compounds significantly alter the normal capacitation process from 30 min of exposure. An increase in ROS production was observed by flow cytometry and considerable DNA fragmentation by the comet assay. The immunocytochemistry showed a decrease of tyrosine phosphorylation in proteins of the equatorial and acrosomal zone of the spermatozoa head. In conclusion, PFOS and PFHxS have toxic effects on the sperm, causing mortality and altering vital parameters for proper sperm capacitation. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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9 pages, 370 KiB  
Article
Efficiency of Tris-Based Extender Steridyl for Semen Cryopreservation in Stallions
by Elena Nikitkina, Artem Musidray, Anna Krutikova, Polina Anipchenko, Kirill Plemyashov and Gennadiy Shiryaev
Animals 2020, 10(10), 1801; https://doi.org/10.3390/ani10101801 - 4 Oct 2020
Cited by 5 | Viewed by 2680
Abstract
The fertilizing ability of stallion sperm after freezing is lower than in other species. The search for the optimal extender, combination of extenders, and the freezing protocol is relevant. The aim of this study was to compare lactose-chelate-citrate-yolk (LCCY) extender, usually used in [...] Read more.
The fertilizing ability of stallion sperm after freezing is lower than in other species. The search for the optimal extender, combination of extenders, and the freezing protocol is relevant. The aim of this study was to compare lactose-chelate-citrate-yolk (LCCY) extender, usually used in Russia, and Steridyl® (Minitube) for freezing sperm of stallions. Steridyl is a concentrated extender medium for freezing ruminant semen. It already contains sterilized egg yolk. Semen was collected from nine stallions, aged from 7 to 12 years old. The total and progressive motility of sperm frozen in Steridyl was significantly higher than in semen frozen in LCCY. The number of spermatozoa with normal morphology in samples frozen in LCCY was 60.4 ± 1.72%, and with Steridyl, 72.4 ± 2.10% (p < 0.01). Semen frozen in Steridyl showed good stimulation of respiration by 2.4-DNP, which indicates that oxidative phosphorylation was retained after freezing–thawing. No differences among the extenders were seen with the DNA integrity of spermatozoa. Six out of ten (60%) mares were pregnant after artificial insemination (AI) by LCCY frozen semen, and 9/12 (75%) by Steridyl frozen semen. No differences among extenders were seen in pregnancy rate. In conclusion, Steridyl was proven to be a good diluent for freezing stallion semen, even though it was developed for ruminants. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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9 pages, 454 KiB  
Article
Semen Quality of Rasa Aragonesa Rams Carrying the FecXR Allele of the BMP15 Gene
by José Alfonso Abecia, Ángel Macías, Adriana Casao, Clara Burillo, Elena Martín, Rosaura Pérez-Pé and Adolfo Laviña
Animals 2020, 10(9), 1628; https://doi.org/10.3390/ani10091628 - 11 Sep 2020
Viewed by 2516
Abstract
The FecXR mutation is a variant of the ovine gene BMP15 in the Rasa Aragonesa breed. Information on the physiological importance of carrying the FecX polymorphism in rams is limited. The aim of this study was to compare semen quality, testicle characteristics, and [...] Read more.
The FecXR mutation is a variant of the ovine gene BMP15 in the Rasa Aragonesa breed. Information on the physiological importance of carrying the FecX polymorphism in rams is limited. The aim of this study was to compare semen quality, testicle characteristics, and fertility rate of rams that carry the FecXR allele. Rams (n = 15) were either FecXR allele carriers (n = 10) or non-carriers, wild type (++) (n = 5). FecXR rams exhibited higher mass motility (p < 0.05), proportion of rapid sperm (p < 0.05), and a lower proportion of slow sperm (p < 0.0001) than did ++ rams. The presence of the FecXR allele was not associated with mean scrotal circumference or testicular length and diameter, although season had a significant (p < 0.05) effect on these traits. Genotype (p < 0.05) and season (p < 0.01) had a significant effect on mean fertility rate, FecXR rams had a higher proportion of pregnant ewes than did ++ rams (p < 0.05). In conclusion, the FecXR allele produced high-quality semen throughout the year, and corresponded with an improvement in some sperm parameters, particularly, mass motility and the proportion of rapid sperm. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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18 pages, 1552 KiB  
Article
The Presence of D-Penicillamine during the In Vitro Capacitation of Stallion Spermatozoa Prolongs Hyperactive-Like Motility and Allows for Sperm Selection by Thermotaxis
by Sara Ruiz-Díaz, Ivan Oseguera-López, David De La Cuesta-Díaz, Belén García-López, Consuelo Serres, Maria José Sanchez-Calabuig, Alfonso Gutiérrez-Adán and Serafin Perez-Cerezales
Animals 2020, 10(9), 1467; https://doi.org/10.3390/ani10091467 - 21 Aug 2020
Cited by 9 | Viewed by 2932
Abstract
Assisted reproductive technologies (ARTs) in the horse still yield suboptimal results in terms of pregnancy rates. One of the reasons for this is the lack of optimal conditions for the sperm capacitation in vitro. This study assesses the use of synthetic human tubal [...] Read more.
Assisted reproductive technologies (ARTs) in the horse still yield suboptimal results in terms of pregnancy rates. One of the reasons for this is the lack of optimal conditions for the sperm capacitation in vitro. This study assesses the use of synthetic human tubal fluid (HTF) supplemented with D-penicillamine (HTF + PEN) for the in vitro capacitation of frozen/thawed stallion spermatozoa by examining capacitation-related events over 180 min of incubation. Besides these events, we explored the in vitro capacity of the spermatozoa to migrate by thermotaxis and give rise to a population of high-quality spermatozoa. We found that HTF induced higher levels of hyperactive-like motility and protein tyrosine phosphorylation (PTP) compared to the use of a medium commonly used in this species (Whitten’s). Also, HTF + PEN was able to maintain this hyperactive-like motility, otherwise lost in the absence of PEN, for 180 min, and also allowed for sperm selection by thermotaxis in vitro. Remarkably, the selected fraction was enriched in spermatozoa showing PTP along the whole flagellum and lower levels of DNA fragmentation when compared to the unselected fraction (38% ± 11% vs 4.4% ± 1.1% and 4.2% ± 0.4% vs 11% ± 2% respectively, t-test p < 0.003, n = 6). This procedure of in vitro capacitation of frozen/thawed stallion spermatozoa in HTF + PEN followed by in vitro sperm selection by thermotaxis represents a promising sperm preparation strategy for in vitro fertilization and intracytoplasmic sperm injection in this species. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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15 pages, 1281 KiB  
Article
Rederivation by Cryopreservation of a Paternal Line of Rabbits Suggests Exhaustion of Selection for Post-Weaning Daily Weight Gain after 37 Generations
by Jorge Daniel Juarez, Francisco Marco-Jiménez, Raquel Lavara and José Salvador Vicente
Animals 2020, 10(8), 1436; https://doi.org/10.3390/ani10081436 - 17 Aug 2020
Cited by 6 | Viewed by 2486
Abstract
Rabbit selection programmes have mainly been evaluated using unselected or divergently selected populations, or populations rederived from cryopreserved embryos after a reduced number of generations. Nevertheless, unselected and divergent populations do not avoid genetic drift, while rederived animals seem to influence phenotypic traits [...] Read more.
Rabbit selection programmes have mainly been evaluated using unselected or divergently selected populations, or populations rederived from cryopreserved embryos after a reduced number of generations. Nevertheless, unselected and divergent populations do not avoid genetic drift, while rederived animals seem to influence phenotypic traits such as birth and adult weights or prolificacy. The study aimed to evaluate the effect of a long-term selection for post-weaning average daily weight gain (ADG) over 37 generations with two rederived populations. Specifically, two coetaneous populations were derived from vitrified embryos with 18 generational intervals (R19 and R37), reducing or avoiding genetic drift and environmental and cryopreservation effects. After two generations of both rederived populations (R21 vs. R39 generations), all evaluated traits showed some progress as a result of the selection, the response being 0.113 g/day by generation. This response does not seem to affect the estimated Gompertz growth curve parameters in terms of the day, the weight at the inflexion point or the adult weight. Moreover, a sexual dimorphism favouring females was observed in this paternal line. Results demonstrated that the selection programme had improved ADG without variations in adult body weight but, after 37 generations of selection, this trait seems exhausted. Given the reduction in the cumulative reproductive performance and as a consequence in the selection pressure, or possibly/perhaps due to an unexpected effect, rederivation could be the cause of this weak selection response observed from generation 18 onwards. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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14 pages, 1137 KiB  
Article
Reproductive Outcomes and Endocrine Profile in Artificially Inseminated versus Embryo Transferred Cows
by Jordana S. Lopes, Estefanía Alcázar-Triviño, Cristina Soriano-Úbeda, Meriem Hamdi, Sebastian Cánovas, Dimitrios Rizos and Pilar Coy
Animals 2020, 10(8), 1359; https://doi.org/10.3390/ani10081359 - 6 Aug 2020
Cited by 14 | Viewed by 3703
Abstract
The increasing use of in vitro embryo production (IVP) followed by embryo transfer (ET), alongside with cryopreservation of embryos, has risen concerns regarding the possible altered pregnancy rates, calving or even neonatal mortality. One of the hypotheses for these alterations is the current [...] Read more.
The increasing use of in vitro embryo production (IVP) followed by embryo transfer (ET), alongside with cryopreservation of embryos, has risen concerns regarding the possible altered pregnancy rates, calving or even neonatal mortality. One of the hypotheses for these alterations is the current culture conditions of the IVP. In an attempt to better mimic the physiological milieu, embryos were produced with female reproductive fluids (RF) as supplements to culture medium, and another group of embryos were supplemented with bovine serum albumin (BSA) as in vitro control. Embryos were cryopreserved and transferred while, in parallel, an in vivo control (artificial insemination, AI) with the same bull used for IVP was included. An overview on pregnancy rates, recipients’ hormonal levels, parturition, and resulting calves were recorded. Results show much similarity between groups in terms of pregnancy rates, gestation length and calves’ weight. Nonetheless, several differences on hormonal levels were noted between recipients carrying AI embryos especially when compared to BSA. Some calving issues and neonatal mortality were observed in both IVP groups. In conclusion, most of the parameters studied were similar between both types of IVP derived embryos and the in vivo-derived embryos, suggesting that the IVP technology used was efficient enough for the safe production of calves. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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12 pages, 284 KiB  
Article
Study of the Metabolomics of Equine Preovulatory Follicular Fluid: A Way to Improve Current In Vitro Maturation Media
by Pablo Fernández-Hernández, María Jesús Sánchez-Calabuig, Luis Jesús García-Marín, María J. Bragado, Alfonso Gutiérrez-Adán, Óscar Millet, Chiara Bruzzone, Lauro González-Fernández and Beatriz Macías-García
Animals 2020, 10(5), 883; https://doi.org/10.3390/ani10050883 - 19 May 2020
Cited by 11 | Viewed by 2977
Abstract
Production of equine embryos in vitro is currently a commercial technique and a reliable way of obtaining offspring. In order to produce those embryos, immature oocytes are retrieved from postmortem ovaries or live mares by ovum pick-up (OPU), matured in vitro (IVM), fertilized [...] Read more.
Production of equine embryos in vitro is currently a commercial technique and a reliable way of obtaining offspring. In order to produce those embryos, immature oocytes are retrieved from postmortem ovaries or live mares by ovum pick-up (OPU), matured in vitro (IVM), fertilized by intracytoplasmic sperm injection (ICSI), and cultured until day 8–10 of development. However, at best, roughly 10% of the oocytes matured in vitro and followed by ICSI end up in successful pregnancy and foaling, and this could be due to suboptimal IVM conditions. Hence, in the present work, we aimed to elucidate the major metabolites present in equine preovulatory follicular fluid (FF) obtained from postmortem mares using proton nuclear magnetic resonance spectroscopy (1H-NMR). The results were contrasted against the composition of the most commonly used media for equine oocyte IVM: tissue culture medium 199 (TCM-199) and Dulbecco’s modified eagle medium/nutrient mixture F-12 Ham (DMEM/F-12). Twenty-two metabolites were identified in equine FF; among these, nine of them are not included in the composition of DMEM/F-12 or TCM-199 media, including (mean ± SEM): acetylcarnitine (0.37 ± 0.2 mM), carnitine (0.09 ± 0.01 mM), citrate (0.4 ± 0.04 mM), creatine (0.36 ± 0.14 mM), creatine phosphate (0.36 ± 0.05 mM), fumarate (0.05 ± 0.007 mM), glucose-1-phosphate (6.9 ± 0.4 mM), histamine (0.25 ± 0.01 mM), or lactate (27.3 ± 2.2 mM). Besides, the mean concentration of core metabolites such as glucose varied (4.3 mM in FF vs. 5.55 mM in TCM-199 vs. 17.5 mM in DMEM/F-12). Hence, our data suggest that the currently used media for equine oocyte IVM can be further improved. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)

Review

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16 pages, 1622 KiB  
Review
The Current Trends in Using Nanoparticles, Liposomes, and Exosomes for Semen Cryopreservation
by Islam M. Saadeldin, Wael A. Khalil, Mona G. Alharbi and Seok Hee Lee
Animals 2020, 10(12), 2281; https://doi.org/10.3390/ani10122281 - 3 Dec 2020
Cited by 45 | Viewed by 5341
Abstract
Cryopreservation is an essential tool to preserve sperm cells for zootechnical management and artificial insemination purposes. Cryopreservation is associated with sperm damage via different levels of plasma membrane injury and oxidative stress. Nanoparticles are often used to defend against free radicals and oxidative [...] Read more.
Cryopreservation is an essential tool to preserve sperm cells for zootechnical management and artificial insemination purposes. Cryopreservation is associated with sperm damage via different levels of plasma membrane injury and oxidative stress. Nanoparticles are often used to defend against free radicals and oxidative stress generated through the entire process of cryopreservation. Recently, artificial or natural nanovesicles including liposomes and exosomes, respectively, have shown regenerative capabilities to repair damaged sperm during the freeze–thaw process. Exosomes possess a potential pleiotropic effect because they contain antioxidants, lipids, and other bioactive molecules regulating and repairing spermatozoa. In this review, we highlight the current strategies of using nanoparticles and nanovesicles (liposomes and exosomes) to combat the cryoinjuries associated with semen cryopreservation. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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24 pages, 1362 KiB  
Review
Oocyte Selection for In Vitro Embryo Production in Bovine Species: Noninvasive Approaches for New Challenges of Oocyte Competence
by Luis Aguila, Favian Treulen, Jacinthe Therrien, Ricardo Felmer, Martha Valdivia and Lawrence C Smith
Animals 2020, 10(12), 2196; https://doi.org/10.3390/ani10122196 - 24 Nov 2020
Cited by 25 | Viewed by 8360
Abstract
The efficiency of producing embryos using in vitro technologies in livestock species rarely exceeds the 30–40% threshold, indicating that the proportion of oocytes that fail to develop after in vitro fertilization and culture is considerably large. Considering that the intrinsic quality of the [...] Read more.
The efficiency of producing embryos using in vitro technologies in livestock species rarely exceeds the 30–40% threshold, indicating that the proportion of oocytes that fail to develop after in vitro fertilization and culture is considerably large. Considering that the intrinsic quality of the oocyte is one of the main factors affecting blastocyst yield, the precise identification of noninvasive cellular or molecular markers that predict oocyte competence is of major interest to research and practical applications. The aim of this review was to explore the current literature on different noninvasive markers associated with oocyte quality in the bovine model. Apart from some controversial findings, the presence of cycle-related structures in ovaries, a follicle size between 6 and 10 mm, large number of surrounding cumulus cells, slightly expanded investment without dark areas, large oocyte diameter (>120 microns), dark cytoplasm, and the presence of a round and smooth first polar body have been associated with better competence. In addition, the combination of oocyte and zygote selection via brilliant cresyl blue (BCB) test, spindle imaging, and the anti-Stokes Raman scattering microscopy together with studies decoding molecular cues in oocyte maturation have the potential to further optimize the identification of oocytes with better developmental competence for in-vitro-derived technologies in livestock species. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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17 pages, 974 KiB  
Review
Extracellular Vesicles, the Road toward the Improvement of ART Outcomes
by Maria G. Gervasi, Ana J. Soler, Lauro González-Fernández, Marco G. Alves, Pedro F. Oliveira and David Martín-Hidalgo
Animals 2020, 10(11), 2171; https://doi.org/10.3390/ani10112171 - 21 Nov 2020
Cited by 11 | Viewed by 3710
Abstract
Nowadays, farm animal industries use assisted reproductive technologies (ART) as a tool to manage herds’ reproductive outcomes, for a fast dissemination of genetic improvement as well as to bypass subfertility issues. ART comprise at least one of the following procedures: collection and handling [...] Read more.
Nowadays, farm animal industries use assisted reproductive technologies (ART) as a tool to manage herds’ reproductive outcomes, for a fast dissemination of genetic improvement as well as to bypass subfertility issues. ART comprise at least one of the following procedures: collection and handling of oocytes, sperm, and embryos in in vitro conditions. Therefore, in these conditions, the interaction with the oviductal environment of gametes and early embryos during fertilization and the first stages of embryo development is lost. As a result, embryos obtained in in vitro fertilization (IVF) have less quality in comparison with those obtained in vivo, and have lower chances to implant and develop into viable offspring. In addition, media currently used for IVF are very similar to those empirically developed more than five decades ago. Recently, the importance of extracellular vesicles (EVs) in the fertility process has flourished. EVs are recognized as effective intercellular vehicles for communication as they deliver their cargo of proteins, lipids, and genetic material. Thus, during their transit through the female reproductive tract both gametes, oocyte and spermatozoa (that previously encountered EVs produced by male reproductive tract) interact with EVs produced by the female reproductive tract, passing them important information that contributes to a successful fertilization and embryo development. This fact highlights that the reproductive tract EVs cargo has an important role in reproductive events, which is missing in current ART media. This review aims to recapitulate recent advances in EVs functions on the fertilization process, highlighting the latest proposals with an applied approach to enhance ART outcome through EV utilization as an additive to the media of current ART procedures. Full article
(This article belongs to the Special Issue The Era of Assisted Reproductive Technologies Tailored to the Specific Necessities of Species, Industry and Case Reports)
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