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Animals
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Development and Application of Assisted Reproductive Techniques in Livestock and...
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Development and Application of Assisted Reproductive Techniques in Livestock and Companion Animals

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

Deadline for manuscript submissions: 31 May 2026 | Viewed by 7931

Special Issue Editors

Dr. Szilárd Bodó

E-Mail Website
Guest Editor
Department of Precision Livestock Farming and Animal Biotechnics, Institute of Animal Sciences, Hungarian University of Agriculture and Life Sciences, Kaposvár Campus, Guba Sándor Street 40, H-7400 Kaposvár, Hungary
Interests: in vitro embryo production; preimplantation genetic diagnosis; micromanipulation; cryopreservation; somatic nuclear transfer; assisted reproductive techniques; animal biotechnology
Dr. Vass Nóra

E-Mail Website
Guest Editor
Department of Animal Nutrition and Physiology, Faculty of Agriculture and Food Sciences and Environmental Management, University of Debrecen, Böszörményi Street 138, H-4032 Debrecen, Hungary
Interests: small ruminant assisted reproduction; farm animal reproduction and reproductive physiology; small ruminant artificial insemination; precision livestock techniques
Dr. Gabriella Kútvölgyi

E-Mail Website
Guest Editor
Department of Precision Livestock Farming and Animal Biotechnics, Institute of Animal Sciences, Hungarian University of Agriculture and Life Sciences, Kaposvár Campus, Guba Sándor Street 40, H-7400 Kaposvár, Hungary
Interests: reproductive biology; equine and small ruminant assisted reproduction; farm animal biotechnology; embryo transfer; sperm cryopreservation; precision livestock techniques; gene conservation
Dr. Tamas Somfai

E-Mail Website
Guest Editor
National Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba 305-8634, Japan
Interests: swine in vitro fertilization; oocyte cryopreservation; gene conservation; reproductive biotechnology; vitrification

Special Issue Information

Dear Colleagues,

The Special Issue will present assisted reproduction techniques currently used and under development in livestock and companion animals. In addition to presenting the results of research on these techniques, we can also give an idea of the economic rationale behind their use. How can these techniques be used for gene conservation in local breeds? Which of the assisted reproduction techniques are likely to make significant breakthroughs? From an animal welfare perspective, how can animal stress and pain levels be effectively reduced during procedures? Can the use of cell nuclear transfer cloning be integrated into animal breeding practice? Are there new methods for gamete and embryo storage? What is the status of the use of sex sorted semen in different species? For which model experiments can the animal in vitro embryo production model be used? What is the current status and potential of preimplantation genetic testing for genomic breeding value determination and genomic studies in different species. What opportunities does genome editing offer for animal breeding? The Special Issue includes review articles and research publications in this field.

Dr. Szilárd Bodó
Dr. Vass Nóra
Dr. Gabriella Kútvölgyi
Dr. Tamas Somfai
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 250 words) can be sent to the Editorial Office for assessment.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Animals is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2400 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • assisted reproduction
  • gene conservation
  • preimplantation genetic testing
  • sex sorted semen
  • animal welfare and reproduction
  • in vitro embryo production

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Published Papers (5 papers)

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Research

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16 pages, 1581 KB  
Article
Breed and Season: Key Determinants of Efficiency in Large-Scale Commercial In Vitro Sheep Embryo Production
by Yubing Wang, Ke Li, Jia Hao, Dayong Chen, Lei Cheng, Huijie He, Riga Wu, Yingjie Wu, Jianhui Tian and Guangyin Xi
Animals 2025, 15(22), 3354; https://doi.org/10.3390/ani15223354 - 20 Nov 2025
Viewed by 1198
Abstract
The efficiency of the laparoscopic ovum pick-up and in vitro embryo production (LOPU-IVEP) system in sheep is governed by a complex interplay of genetic and environmental factors. This study comprehensively evaluated the effects of breed, season, and their interaction on oocyte retrieval, embryonic [...] Read more.
The efficiency of the laparoscopic ovum pick-up and in vitro embryo production (LOPU-IVEP) system in sheep is governed by a complex interplay of genetic and environmental factors. This study comprehensively evaluated the effects of breed, season, and their interaction on oocyte retrieval, embryonic development, and pregnancy outcomes in a commercial setting. A 20-month longitudinal analysis of a large commercial cohort revealed that breed was the dominant factor determining oocyte quantity, whereas season exerted a stronger influence on developmental competence. Specifically, autumn and winter provided optimal conditions, resulting in significantly higher cleavage rate and blastocyst rate (p < 0.05). A significant breed-by-season interaction further identified synergistic combinations, such as Black-headed Suffolk in autumn or winter, which achieved superior embryo production outcomes. Importantly, correlation analysis revealed that oocyte quantity was not predictive of developmental potential or pregnancy success. These findings underscore the necessity of a precision management strategy that aligns LOPU schedules with breed-specific seasonal advantages to maximize annual embryo production efficiency. Full article
(This article belongs to the Special Issue Development and Application of Assisted Reproductive Techniques in Livestock and Companion Animals)
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11 pages, 212 KB  
Article
Establishment of a Single-Oocyte Culture System for Pigs and Its Validation Using Curcumin as a Model Antioxidant for Oocyte Maturation
by Zhao Namula, Takeshige Otoi, Theerawat Tharasanit, Kaywalee Chatdarong, Megumi Nagahara, Oky Setyo Widodo, Aya Nakai, Suong Thi Nguyen, Yuichiro Nakayama, Maki Hirata and Fuminori Tanihara
Animals 2025, 15(22), 3295; https://doi.org/10.3390/ani15223295 - 14 Nov 2025
Viewed by 723
Abstract
Since individual embryos cannot be evaluated in group culture, establishing a single culture from in vitro maturation to in vitro culture may provide new insights into oocyte and embryo quality. This study aimed to develop a single culture system for individual oocytes, from [...] Read more.
Since individual embryos cannot be evaluated in group culture, establishing a single culture from in vitro maturation to in vitro culture may provide new insights into oocyte and embryo quality. This study aimed to develop a single culture system for individual oocytes, from in vitro maturation through fertilization to embryo development. The effects of curcumin supplementation during in vitro maturation on oocyte maturation, embryo development, and embryo quality were examined in single and group culture systems. Porcine oocytes were cultured individually in 20 µL microdroplets, with one oocyte per droplet, or in groups of 50 oocytes per 500 µL. The maturation medium contained curcumin at concentrations of 20 µM or less. Supplementation with 10 µM curcumin increased oocyte maturation in both systems compared to the controls. The fertilization rates and oocyte/embryo quality did not differ among the treatment groups. Oocytes matured with 10 µM curcumin in a single culture showed a higher blastocyst formation rate (7.0%) than the control (2.3%). In the group culture, 10 µM curcumin increased cleavage rates compared to the control (75.2% vs. 63.0%), but blastocyst formation rates did not differ. Blastocyst formation rates were similar between single and group cultures under control (2.3% and 4.3%, respectively) or 10 µM curcumin (7.0% and 11.4%, respectively) conditions. Therefore, porcine oocytes can develop to the blastocyst stage in a single culture system. Incorporating antioxidants during in vitro maturation may be an effective condition for in vitro embryo culture that can be implemented in a single oocyte. Full article
(This article belongs to the Special Issue Development and Application of Assisted Reproductive Techniques in Livestock and Companion Animals)
14 pages, 5330 KB  
Article
Development of IVF Porcine Embryos in Microwell Culture System
by Kayode Balogun and Zoltan Machaty
Animals 2025, 15(17), 2528; https://doi.org/10.3390/ani15172528 - 28 Aug 2025
Viewed by 1882
Abstract
We investigated the development of IVF porcine embryos incubated in microwells of Well-of-the-Well (WOW) culture dishes. In Experiment 1, 500 embryos were cultured under five different culture conditions: 10 individual embryos in microwells covered by 50 μL PZM-3 medium (50 μL WOW); 10 [...] Read more.
We investigated the development of IVF porcine embryos incubated in microwells of Well-of-the-Well (WOW) culture dishes. In Experiment 1, 500 embryos were cultured under five different culture conditions: 10 individual embryos in microwells covered by 50 μL PZM-3 medium (50 μL WOW); 10 embryos together in a well of 50 μL PZM-3 (50 μL Well); 10 embryos in microwells covered by 20 μL PZM-3 (20 μL WOW); 10 embryos in a well of 20 μL PZM-3 (20 μL Well); and 10 embryos in a 20 μL drop of PZM-3 in traditional dishes (control). In Experiment 2, the best-performing treatment group from Experiment 1 (50 μL WOW) was further compared to control using a total of 350 embryos. There was no difference in the percentage of cleaved embryos among the groups in both experiments. In Experiment 1, the percentage of blastocysts from 50 μL WOW (42.00 ± 6.29%) was higher (p = 0.001) than in 50 μL Well (29.00 ± 4.58%), 20 μL WOW (13.00 ± 2.13%), 20 μL Well (20.00 ± 2.98%) and control (25.00 ± 5.22%). Similarly, in Experiment 2, blastocyst formation and nuclei number in the blastocysts from 50 μL WOW (37.86 ± 3.95% and 38.97 ± 1.80) were higher (p = 0.040 and p = 0.017) than in the control (28.10 ± 2.64% and 33.21 ± 1.56). The results indicate that individual culture in microwells improves the development of IVF pig embryos. Full article
(This article belongs to the Special Issue Development and Application of Assisted Reproductive Techniques in Livestock and Companion Animals)
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14 pages, 475 KB  
Article
Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and Foals
by Jenin V. Cortez, Kylie Hardwicke, Carlos E. Méndez-Calderón and Christopher G. Grupen
Animals 2025, 15(13), 1961; https://doi.org/10.3390/ani15131961 - 3 Jul 2025
Cited by 1 | Viewed by 2372
Abstract
The asynchrony of cytoplasmic and nuclear maturation in cumulus–oocyte complexes (COCs) due to prematurely declining concentrations of cyclic adenosine monophosphate (cAMP) has been shown to result in reduced oocyte developmental competence. The objective of this study was to evaluate the effect of pre-IVM [...] Read more.
The asynchrony of cytoplasmic and nuclear maturation in cumulus–oocyte complexes (COCs) due to prematurely declining concentrations of cyclic adenosine monophosphate (cAMP) has been shown to result in reduced oocyte developmental competence. The objective of this study was to evaluate the effect of pre-IVM treatment with cAMP modulators for different durations on the developmental potential of equine oocytes used for cloned embryo production. Collected COCs were transferred to cryovials filled with transport medium at 20–22 °C. Within the cryovials, the COCs were either untreated (Control) for 18 h or treated with 50 µM forskolin and 100 µM 3-isobutyl-1-methylxanthine for the first 4 h (Pre-IVM 4 h) or the entire 18 h (Pre-IVM 18 h). Oocytes were then transferred to maturation medium and incubated for a further 22–24 h at 38.5 °C in 5% CO2 in air. Somatic cell nuclear transfer embryos were then produced using the meiotically mature oocytes and donor cells from six different fibroblast cell lines. The rates of maturation and embryo development did not differ significantly between the groups, though blastocyst formation tended to be inferior in the Pre-IVM 4 h group compared with the Control group (p = 0.06). Of 67 blastocysts produced, 23 were transferred to recipient mares on Day 4 or 5 post-ovulation. Regarding the pregnancy outcomes, no significant differences were found between the groups, and four viable foals were born, each derived from a different donor cell line. The findings expand on those from previous evaluations of this biphasic IVM system, and indicate that the cAMP-modulating treatments exert limited effects under the pre-IVM conditions used here. Full article
(This article belongs to the Special Issue Development and Application of Assisted Reproductive Techniques in Livestock and Companion Animals)
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Review

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29 pages, 2290 KB  
Review
Impact of Nutrition on Embryo Production in Cattle: Mechanistic Insights
by Ramanathan Kasimanickam and Vanmathy Kasimanickam
Animals 2026, 16(6), 892; https://doi.org/10.3390/ani16060892 - 12 Mar 2026
Viewed by 1042
Abstract
Reproductive efficiency in cattle is critically dependent on embryo quantity and quality, particularly in assisted reproductive technology (ART) programs such as superovulation, embryo transfer, and embryo production. Nutrition is a key determinant of embryo yield through its regulatory effects on metabolic signaling, ovarian [...] Read more.
Reproductive efficiency in cattle is critically dependent on embryo quantity and quality, particularly in assisted reproductive technology (ART) programs such as superovulation, embryo transfer, and embryo production. Nutrition is a key determinant of embryo yield through its regulatory effects on metabolic signaling, ovarian function, oocyte competence, and early embryogenesis. This review synthesizes the current evidence describing mechanistic links between nutritional status and embryo production in dairy and beef cattle across both in vivo and in vitro systems. Energy balance, protein supply, micronutrients, and fatty acids influence metabolic hormones including insulin, insulin-like growth factor-1, and leptin, which regulate hypothalamic–pituitary–gonadal axis activity, follicular recruitment, and steroidogenesis. Negative energy balance disrupts endocrine signaling, elevates circulating non-esterified fatty acids, increases oxidative stress, and impairs oocyte mitochondrial function, resulting in reduced embryo yield, compromised blastocyst quality, and diminished cryotolerance. Targeted micronutrients such as selenium, zinc, vitamins A and E, B-complex vitamins, and omega-3 fatty acids enhance antioxidant capacity, membrane integrity, and epigenetic regulation, thereby supporting embryo viability and post-transfer survival. Furthermore, early-life nutrition programs long-term reproductive capacity by influencing ovarian reserve establishment and oocyte epigenetic competence. Strategic nutritional management is therefore essential to optimize ART outcomes and promote sustainable genetic progress in cattle production systems. Full article
(This article belongs to the Special Issue Development and Application of Assisted Reproductive Techniques in Livestock and Companion Animals)
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