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Animals
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In Vitro Embryo Production in Ruminants
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In Vitro Embryo Production in Ruminants

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

Deadline for manuscript submissions: closed (31 December 2021) | Viewed by 23964

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

Special Issue Editors

Dr. Ignacio Contreras

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Guest Editor
Dipartimento di Medicina Veterinaria, Università degli Studi di Sassari, Sassari, Italy
Interests: embryo development; maternal–embryo relationships; ovarian function; estrus synchronization
Special Issues, Collections and Topics in MDPI journals
Dr. Sandra Soto

E-Mail Website
Guest Editor
University of Illinois Urbana-Champaign, Urbana, IL, USA
Interests: embryo development; oocyte; sperm reservoir; melatonin; biphasic in vitro maturation; in vitro fertilization

Special Issue Information

Dear Colleagues,

Assisted reproductive technologies applied to ruminants, such as multiovulation, artificial insemination, embryo transfer, and in vitro fertilization (IVF), have been useful tools to accelerate the genetic progress in these species. Despite the effort to improve the IVF outcome in small and large ruminants in terms of embryo yield and quality, there are still many challenges to overcome. Further optimization of these technologies will potentially lead to their widespread application in ruminant breeding programs. This includes enhancing oocyte and sperm selection procedures, as well as designing optimal culture media and protocols for oocyte maturation, IVF, and embryo culture. Thus, the aim of this Special Issue is to highlight state-of-the-art advances in IVF—including review and research articles—applied to small and large ruminants. Finally, we hope that this Special Issue will raise interest in the fascinating field of IVF.

Dr. Ignacio Contreras
Dr. Sandra Soto
Guest Editors

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Keywords

  • oocyte
  • in vitro maturation
  • in vitro fertilization
  • blastocyst yield
  • small ruminants
  • large ruminants

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Published Papers (5 papers)

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Research

Jump to: Review, Other

13 pages, 1706 KiB  
Article
Establishment of a Wisent (Bison bonasus) Germplasm Bank
by Anna Maria Duszewska, Magdalena Baraniewicz-Kołek, Jarosław Wojdan, Katarzyna Barłowska, Wojciech Bielecki, Paweł Gręda, Wojciech Niżański and Wanda Olech
Animals 2022, 12(10), 1239; https://doi.org/10.3390/ani12101239 - 11 May 2022
Cited by 5 | Viewed by 2171
Abstract
The wisent, or European bison (Bison bonasus), belongs to the same family (Bovidae) as the American bison and domestic cattle. The wisent is the largest mammal in Europe, and is called the “Forest Emperor”. The wisent is listed as “Vulnerable” on [...] Read more.
The wisent, or European bison (Bison bonasus), belongs to the same family (Bovidae) as the American bison and domestic cattle. The wisent is the largest mammal in Europe, and is called the “Forest Emperor”. The wisent is listed as “Vulnerable” on the IUCN Red List, and is protected by international law. Achievements in reproductive biotechnology have opened new possibilities for the cryoconservation of the wisent germplasm. Therefore, this research aimed to improve a strategy for the protection and preservation of the European bison through the creation of a wisent germplasm bank, based on the following procedures: isolation and in vitro maturation (IVM) of oocytes, in vitro fertilization (IVF) of matured oocytes, in vitro embryo culture (IVC), and embryo cryopreservation. Wisent ovaries were isolated from females outside the reproductive season, and eliminated from breeding for reasons other than infertility. Cumulus–oocyte complexes (COCs) were isolated from follicles greater than 2 mm in diameter and matured for 24 h and 30 h. After IVM, COCs were fertilized in vitro with wisent sperm. The obtained wisent zygotes, based on oocytes matured for 24 h and 30 h, were cultured for 216 h. Embryos at the morula and early blastocyst stages were vitrified and then warmed and transferred to interspecies recipients (Bos taurus). USG and biochemical tests were used to monitor pregnancies. This study obtained embryos in the morula and early blastocyst stages only after oocytes were fertilized and matured for 30 h. On average, per oocyte donor, 12.33 ± 0.5 COCs were isolated, and only 9.33 ± 0.61 COCs were qualified for in vitro maturation (75.68%), while 9.16 ± 0.48 COCs were matured (84.32%). On average, per donor, 5.5 ± 0.34 embryos were cleaved (59.96%) after 48 h post-fertilization (hpf), and 3.33 ± 0.21 achieved the eight-cell stage (36.52%) after 96 hpf, while 1 ± 0.21 morula and early blastocyst stages (10.71%) were achieved after 216 hpf. A total of six embryos (one morula and five early blastocysts) were obtained and vitrified; after warming, five of them were interspecies transferred to cattle (Bos taurus). On day 41 after fertilization, 3 out of 5 pregnancies were detected based on USG, P4, and PAG tests. However, no pregnancy was observed on day 86 after fertilization, indicating embryo resorption. This study shows that obtaining wisent embryos in vitro, and subsequent cryopreservation to create a wisent embryo bank, can be applied and implemented for the wisent protection program. Full article
(This article belongs to the Special Issue In Vitro Embryo Production in Ruminants)
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14 pages, 4018 KiB  
Article
Heat Shock Protein 70 Improves In Vitro Embryo Yield and Quality from Heat Stressed Bovine Oocytes
by Konstantina Stamperna, Themistoklis Giannoulis, Eleni Dovolou, Maria Kalemkeridou, Ioannis Nanas, Katerina Dadouli, Katerina Moutou, Zissis Mamuris and Georgios S. Amiridis
Animals 2021, 11(6), 1794; https://doi.org/10.3390/ani11061794 - 16 Jun 2021
Cited by 15 | Viewed by 3176
Abstract
Heat shock protein 70 (HSP70) is a chaperon that stabilizes unfolded or partially folded proteins, preventing inappropriate inter- and intramolecular interactions. Here, we examined the developmental competence of in vitro matured oocytes exposed to heat stress with or without HSP70. Bovine oocytes were [...] Read more.
Heat shock protein 70 (HSP70) is a chaperon that stabilizes unfolded or partially folded proteins, preventing inappropriate inter- and intramolecular interactions. Here, we examined the developmental competence of in vitro matured oocytes exposed to heat stress with or without HSP70. Bovine oocytes were matured for 24 h at 39 °C without (group C39) or with HSP70 (group H39) and at 41 °C for the first 6 h, followed by 16 h at 39 °C with (group H41) or without HSP70 (group C41). After insemination, zygotes were cultured for 9 days at 39 °C. Cleavage and embryo yield were assessed 48 h post insemination and on days 7, 8, 9, respectively. Gene expression was assessed by RT-PCR in oocytes, cumulus cells and blastocysts. In C41, blastocysts formation rate was lower than in C39 and on day 9 it was lower than in H41. In oocytes, HSP70 enhanced the expression of three HSP genes regardless of incubation temperature. HSP70 at 39 °C led to tight coordination of gene expression in oocytes and blastocysts, but not in cumulus cells. Our results imply that HSP70, by preventing apoptosis, supporting signal transduction, and increasing antioxidant protection of the embryo, protects heat stressed maturing bovine oocyte and restores its developmental competence. Full article
(This article belongs to the Special Issue In Vitro Embryo Production in Ruminants)
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9 pages, 1341 KiB  
Article
Reproductive Seasonality Affects In Vitro Embryo Production Outcomes in Adult Goats
by Joanna M.G. Souza-Fabjan, Lucas F.L. Correia, Ribrio I.T.P. Batista, Yann Locatelli, Vicente J.F. Freitas and Pascal Mermillod
Animals 2021, 11(3), 873; https://doi.org/10.3390/ani11030873 - 18 Mar 2021
Cited by 3 | Viewed by 2754
Abstract
Reproductive seasonality may have a considerable influence on the efficiency of assisted reproductive technologies in seasonal species. This study evaluated the effect of season on cleavage, blastocyst rates and quality of in vitro produced (IVP) goat embryos. In total, 2348 cumulus–oocyte complexes (COCs) [...] Read more.
Reproductive seasonality may have a considerable influence on the efficiency of assisted reproductive technologies in seasonal species. This study evaluated the effect of season on cleavage, blastocyst rates and quality of in vitro produced (IVP) goat embryos. In total, 2348 cumulus–oocyte complexes (COCs) were recovered from slaughterhouse ovaries and subjected to the same IVP system throughout 1.5 years (49 replicates). The odds ratio (OR) among seasons was calculated from values of cleavage and blastocyst rates in each season. Cleavage rate was lower (p < 0.05) in spring (anestrus), in comparison with either autumn (peak of breeding season) or summer, while the winter had intermediate values. Furthermore, lower OR of cleavage was observed in spring. Blastocyst formation rate (from initial number of COCs) was higher (p < 0.05) in autumn (52 ± 2.5%) when compared with the other seasons (combined rates: 40 ± 1.9%). Moreover, its OR was higher (p < 0.05) in autumn compared to all other seasons and impaired in the spring compared to winter (OR: 0.54) and summer (OR: 0.48). Embryo hatchability and blastocyst cell number were similar (p > 0.05) among seasons. In conclusion, the breeding season leads to improved oocyte developmental competence, resulting in higher cleavage and blastocyst yield, whereas embryo quality remained similar throughout the years. Full article
(This article belongs to the Special Issue In Vitro Embryo Production in Ruminants)
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Review

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20 pages, 1173 KiB  
Review
In Vitro Production of Embryos from Prepubertal Holstein Cattle and Mediterranean Water Buffalo: Problems, Progress and Potential
by Luke Currin, Hernan Baldassarre and Vilceu Bordignon
Animals 2021, 11(8), 2275; https://doi.org/10.3390/ani11082275 - 1 Aug 2021
Cited by 19 | Viewed by 9485
Abstract
Laparoscopic ovum pick-up (LOPU) coupled with in vitro embryo production (IVEP) in prepubertal cattle and buffalo accelerates genetic gain. This article reviews LOPU-IVEP technology in prepubertal Holstein Cattle and Mediterranean Water Buffalo. The recent expansion of genomic-assisted selection has renewed interest and demand [...] Read more.
Laparoscopic ovum pick-up (LOPU) coupled with in vitro embryo production (IVEP) in prepubertal cattle and buffalo accelerates genetic gain. This article reviews LOPU-IVEP technology in prepubertal Holstein Cattle and Mediterranean Water Buffalo. The recent expansion of genomic-assisted selection has renewed interest and demand for prepubertal LOPU-IVEP schemes; however, low blastocyst development rates has constrained its widespread implementation. Here, we present an overview of the current state of the technology, limitations that persist and suggest possible solutions to improve its efficiency, with a focus on gonadotropin stimulations strategies to prime oocytes prior to follicular aspiration, and IVEP procedures promoting growth factor metabolism and limiting oxidative and endoplasmic reticulum stress. Full article
(This article belongs to the Special Issue In Vitro Embryo Production in Ruminants)
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Other

Jump to: Research, Review

9 pages, 1130 KiB  
Brief Report
Ultrasonic Cutting of Frozen Semen Straws to Optimize the Use of Spermatozoa for In Vitro Fertilization
by Sung Woo Kim, Jae-Yeong Lee, Bongki Kim, Chan-Lan Kim, In-Sul Hwang, Yeoung-Gyu Ko and Sung-Soo Lee
Animals 2020, 10(11), 2152; https://doi.org/10.3390/ani10112152 - 19 Nov 2020
Cited by 2 | Viewed by 3534
Abstract
The objective of the present study was to establish conditions for using technology that can potentially enhance the efficiency of bovine embryos derived from in vitro fertilization (IVF) with frozen semen. Frozen semen from selected bulls can be stored indefinitely in liquid nitrogen [...] Read more.
The objective of the present study was to establish conditions for using technology that can potentially enhance the efficiency of bovine embryos derived from in vitro fertilization (IVF) with frozen semen. Frozen semen from selected bulls can be stored indefinitely in liquid nitrogen as genetic resources; however, these resources are considered consumable because they cannot be regenerated. Therefore, to optimize the utilization of frozen semen, as many oocytes as possible should be fertilized with one straw. However, a sufficient number of prepared oocytes might not be available for one experiment, which can limit the use of the total spermatozoa population. Thus, an economical method for producing embryos needs to be established by optimizing technology for transplantable embryos. In this study, the utilization of frozen semen was increased by dividing the straw with an ultrasonic cutter. The post-thaw survival rate of uncut straws from Korean Proven Bulls did not differ from that of half cuttings. When ultrasonic cutting was applied to frozen semen, spermatozoa could be prepared for IVF trials at least four times, and blastocysts were produced. Therefore, cutting frozen semen with an ultrasonic cutter represents a potentially useful tool to expand genetic resources from excellent breeding stocks. This approach could also be valuable in the field of IVF of endangered species or rare breeds for their preservation, as well as in ovum pick-up (OPU) techniques. Full article
(This article belongs to the Special Issue In Vitro Embryo Production in Ruminants)
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