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Animals
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Challenges in the Reproduction of Equids
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Challenges in the Reproduction of Equids

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

Deadline for manuscript submissions: 1 May 2025 | Viewed by 26537

Special Issue Editor

Prof. Dr. Christine Aurich

E-Mail Website
Guest Editor
Artificial Insemination and Embryo Transfer, Department for Small Animals and Horses, Vetmeduni Vienna, Veterinärplatz 1, 1210 Vienna, Austria
Interests: equine reproduction; gonadal function and dysfunction; early pregnancy; late pregnancy and parturition

Special Issue Information

Dear Colleagues,

Horses are selecting for breeding purposes either because of their performance or appearance, but fertility is often neglected. This contributes to low fertility in equine reproduction. More recently, also fertility problems in donkeys have raised an increasing interest. During the past few decades, studies have focused on better understanding the mechanisms that regulate the reproductive processes in equids. There are, however, still many aspects of equine reproduction that remain unknown or have not yet been proven. Many published studies have answered some of these questions but have also raised new ones, and several of those questions are still unanswered.

This Special Issue aims to improve our understanding of reproduction in equids; therefore, we invite high-quality and original research or review papers that address new insights on this topic, with main focus on the physiology and pathology of equine and donkey reproduction.

Prof. Christine Aurich
Guest Editor

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. 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

  • equine reproduction
  • gonadal function and dysfunction
  • early pregnancy
  • late pregnancy and parturition

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Published Papers (7 papers)

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Research

Jump to: Review

12 pages, 612 KiB  
Article
Optimization of the Equine-Sperm Freeze Test in Purebred Spanish Horses by Incorporating Colloidal Centrifugation
by Luna Gutiérrez-Cepeda, Francisco Crespo, Juan Carlos Blazquez and Consuelo Serres
Animals 2023, 13(3), 382; https://doi.org/10.3390/ani13030382 - 22 Jan 2023
Cited by 5 | Viewed by 2819
Abstract
The Purebred Spanish Horse, according to our clinical experience, is characterized by having a high number of stallions that do not meet the international commercial recommendations for equine-sperm cryopreservation. This means that artificial insemination with frozen semen from these stallions is less widespread [...] Read more.
The Purebred Spanish Horse, according to our clinical experience, is characterized by having a high number of stallions that do not meet the international commercial recommendations for equine-sperm cryopreservation. This means that artificial insemination with frozen semen from these stallions is less widespread than in other breeds. In this study, we investigated if the incorporation of single-layer colloidal centrifugation prior to cryopreservation in clinical conditions could increase the number of ejaculates of Purebred Spanish stallions suitable for this processing, observing the influence of centrifugation and freezing extender protocol on post-thawed sperm motility. Using colloidal centrifugation, the percentage of ejaculates available to be frozen was increased from 35% (6/17) to 71% (12/17), doubling the number of samples that could have been subjected to cryopreservation. We only found significant differences in linearity (LIN) and lateral head displacement (ALH) after 5 min of incubation at 37 °C between colloidal and simple centrifugation processing techniques. No significant differences were found between the two different colloidal protocols in any of the variables considered. Colloidal centrifugation allowed us to obtain, from worse fresh-quality ejaculates, thawed sperm doses with similar quality to that of good-quality ejaculates. BotuCrio® produced, in general, higher motility parameters and its characteristics than the other extenders analyzed, with significant differences found in comparison to Inra-Freeze® and Lac-Edta in both total (MOT) and progressive motility (PMOT) when using colloidal centrifugation and only in PMOT when applying simple centrifugation. Colloidal centrifugation optimized the efficiency of cryopreservation, as it allowed us to increase the number of ejaculates of Purebred Spanish Horses suitable to be frozen. Including these semen processing techniques in the freeze test could help to optimize equine-sperm cryopreservation protocols, especially when dealing with individuals or breeds for which initially low sperm quality prevents or limits their inclusion in sperm cryopreservation programs. Full article
(This article belongs to the Special Issue Challenges in the Reproduction of Equids)
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14 pages, 880 KiB  
Article
Serological and Uterine Biomarkers for Detecting Endometritis in Mares
by Stefano Cecchini Gualandi, Tommaso Di Palma and Raffaele Boni
Animals 2023, 13(2), 253; https://doi.org/10.3390/ani13020253 - 11 Jan 2023
Cited by 2 | Viewed by 2452
Abstract
Serological analysis may provide relevant information on endometritis diagnostics. Therefore, mares scheduled for AI with refrigerated semen, at the time of heat signs, underwent blood and uterine fluid samplings using a swab, uterine lavage for culture analysis, and treatment with human chorionic gonadotropin [...] Read more.
Serological analysis may provide relevant information on endometritis diagnostics. Therefore, mares scheduled for AI with refrigerated semen, at the time of heat signs, underwent blood and uterine fluid samplings using a swab, uterine lavage for culture analysis, and treatment with human chorionic gonadotropin to induce ovulation. After 24–28 h, the mares were inseminated and, if positive at the culture test, treated with antibiotics chosen based on the susceptibility test. Uterine cells obtained by swabs were used for cytological examination with both classical and fluorescence techniques. Blood serum and uterine fluid samples were analyzed for assessing parameters related to redox balance, inflammation, and protease regulator potential. In blood serum, total antioxidant capacity, measured as the ferric reducing ability of plasma (FRAP), was significantly lower in cytologically endometritis-positive than -negative mares. In the uterine fluid, total thiol levels (TTL), nitric oxide metabolites (NOx), protease activity and total protein content varied significantly between groups. Although the cytological examination was more capable of discriminating between endometritis-positive and -negative mares in relation to the parameters examined, no statistically significant differences emerged in terms of pregnancy rate in relation to cytological and culture diagnosis as well as in mares diagnosed as positive and negative for endometritis. Full article
(This article belongs to the Special Issue Challenges in the Reproduction of Equids)
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15 pages, 4051 KiB  
Article
Influence of the Season and Region Factor on Phosphoproteome of Stallion Epididymal Sperm
by Katarzyna Dyrda, Aleksandra Orzołek, Joanna Ner-Kluza and Paweł Wysocki
Animals 2021, 11(12), 3487; https://doi.org/10.3390/ani11123487 - 7 Dec 2021
Cited by 1 | Viewed by 2582
Abstract
Epididymal maturation can be defined as a scope of changes occurring during epididymal transit that prepare spermatozoa to undergo capacitation. One of the most common post-translational modifications involved in the sperm maturation process and their ability to fertilise an oocyte is the phosphorylation [...] Read more.
Epididymal maturation can be defined as a scope of changes occurring during epididymal transit that prepare spermatozoa to undergo capacitation. One of the most common post-translational modifications involved in the sperm maturation process and their ability to fertilise an oocyte is the phosphorylation of sperm proteins. The aim of this study was to compare tyrosine, serine, and threonine phosphorylation patterns of sperm proteins isolated from three subsequent segments of the stallion epididymis, during and out of the breeding season. Intensities of phosphorylation signals and phosphoproteins profiles varied in consecutive regions of the epididymis. However, significant differences in the phosphorylation status were demonstrated in case of endoplasmic reticulum chaperone BiP (75 and 32 kDa), protein disulfide-isomerase A3 (50 kDa), nesprin-1 (23 kDa), peroxiredoxin-5 (17 kDa), and protein bicaudal D homolog (15 kDa) for season x type of phosphorylated residues variables. Significant differences in the phosphorylation status were also demonstrated in case of endoplasmic reticulum chaperone BiP and albumin (61 kDa), protein disulfide-isomerase A3 (50 kDa), and protein bicaudal D homolog (15 kDa) for region x type of phosphorylated residues variables. Full article
(This article belongs to the Special Issue Challenges in the Reproduction of Equids)
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18 pages, 3418 KiB  
Article
Sexual Differentiation and Primordial Germ Cell Distribution in the Early Horse Fetus
by Dragos Scarlet, Stephan Handschuh, Ursula Reichart, Giorgia Podico, Robyn E. Ellerbrock, Sebastián Demyda-Peyrás, Igor F. Canisso, Ingrid Walter and Christine Aurich
Animals 2021, 11(8), 2422; https://doi.org/10.3390/ani11082422 - 17 Aug 2021
Cited by 10 | Viewed by 6463
Abstract
It was the aim of this study to characterize the development of the gonads and genital ducts in the equine fetus around the time of sexual differentiation. This included the identification and localization of the primordial germ cell population. Equine fetuses between 45 [...] Read more.
It was the aim of this study to characterize the development of the gonads and genital ducts in the equine fetus around the time of sexual differentiation. This included the identification and localization of the primordial germ cell population. Equine fetuses between 45 and 60 days of gestation were evaluated using a combination of micro-computed tomography scanning, immunohistochemistry, and multiplex immunofluorescence. Fetal gonads increased in size 23-fold from 45 to 60 days of gestation, and an even greater increase was observed in the metanephros volume. Signs of mesonephros atrophy were detected during this time. Tubular structures of the fetal testes were present from day 50 onwards, whereas cell clusters dominated in the fetal ovary. The genital ducts were well-differentiated and presented a lumen in all samples. No sign of mesonephric or paramesonephric duct degeneration was detected. Expression of AMH was strong in the fetal testes but absent in ovaries. Irrespective of sex, primordial germ cells selectively expressed LIN28. Migration of primordial germ cells from the mesonephros to the gonad was detected at 45 days, but not at 60 days of development. Their number and distribution within the gonad were influenced (p < 0.05) by fetal sex. Most primordial germ cells (86.8 ± 3.2% in females and 84.6 ± 4.7% in males) were characterized as pluripotent according to co-localization with CD117. However, only a very small percentage of primordial germ cells were proliferating (7.5 ± 1.7% in females and 3.2 ± 1.2% in males) based on co-localization with Ki67. It can be concluded that gonadal sexual differentiation in the horse occurs asynchronously with regard to sex but already before 45 days of gestation. Full article
(This article belongs to the Special Issue Challenges in the Reproduction of Equids)
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20 pages, 2178 KiB  
Article
Epigenetic Changes in Equine Embryos after Short-Term Storage at Different Temperatures
by Gustavo D. A. Gastal, Dragos Scarlet, Maria Melchert, Reinhard Ertl and Christine Aurich
Animals 2021, 11(5), 1325; https://doi.org/10.3390/ani11051325 - 6 May 2021
Cited by 1 | Viewed by 2838
Abstract
In embryos subjected to assisted reproductive techniques, epigenetic modifications may occur that can influence embryonic development and the establishment of pregnancy. In horses, the storage temperature during transport of fresh embryos before transfer is a major concern. The aim of this study was, [...] Read more.
In embryos subjected to assisted reproductive techniques, epigenetic modifications may occur that can influence embryonic development and the establishment of pregnancy. In horses, the storage temperature during transport of fresh embryos before transfer is a major concern. The aim of this study was, therefore, to determine the effects of two storage temperatures (5 °C and 20 °C) on equine embryos, collected at day seven after ovulation and stored for 24 h, on: (i) morphological development; (ii) expression of candidate genes associated with embryo growth and development, maternal recognition of pregnancy, methylation and apoptosis, and (iii) gene-specific and global DNA methylation. Embryos (n = 80) were collected on day seven or day eight after ovulation and assigned to four groups: day seven control (E7F, fresh); day seven, stored for 24 h at 5 °C (E5C); day seven, stored for 24 h at 20 °C (E20C) and day eight control (E8F, fresh 24h time control). The embryos and the storage medium (EquiHold, holding medium, Minitube, Tiefenbach, Germany) from all treatment groups were analyzed for (i) medium temperature, pH, and lipid peroxidation (malondialdehyde; MDA) and (ii) embryo morphology, mRNA expression and DNA methylation (immunohistochemistry and gene-specific DNA methylation). The size of embryos stored at 5 °C was larger (p < 0.01), whereas embryos stored at 20 °C were smaller (p < 0.05) after 24 h. There were no changes in pH and MDA accumulation irrespective of the group. The mRNA expression of specific genes related to growth and development (POU5F1, SOX2, NANOG), maternal recognition of pregnancy (CYP19A1, PTGES2), DNA methylation (DNMT1, DNMT3A, DNMT3B) and apoptosis (BAX) in the E5C and E20C were either up or downregulated (p < 0.05) when compared to controls (E7F and E8F). The immune expression of 5mC and 5hmC was similar among treatment groups. Percentage of methylation in the CpG islands was lower in the specific genes ESR1, NANOG and DNMT1 (p < 0.001) in E20C embryos when compared to E8F (advanced embryo stage). Therefore, our study demonstrates for the first time the gene-specific and global DNA methylation status of fresh equine embryos collected on days seven and eight after ovulation. Although our results suggest some beneficial effects of storage at 20 °C in comparison to 5 °C, the short-term storage, regardless of temperature, modified gene expression and methylation of genes involved in embryo development and may compromise embryo viability and development after transfer. Full article
(This article belongs to the Special Issue Challenges in the Reproduction of Equids)
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15 pages, 4613 KiB  
Article
Expression of Enzymes Associated with Prostaglandin Synthesis in Equine Conceptuses
by Sven Budik, Ingrid Walter, Marie-Christine Leitner, Reinhard Ertl and Christine Aurich
Animals 2021, 11(4), 1180; https://doi.org/10.3390/ani11041180 - 20 Apr 2021
Cited by 8 | Viewed by 2913
Abstract
In the horse, mobility of the conceptus is required for maternal recognition of pregnancy depending on secretion of prostaglandins by the conceptus. The aim of this study was to determine the expression and localization of key enzymes of the different pathways leading to [...] Read more.
In the horse, mobility of the conceptus is required for maternal recognition of pregnancy depending on secretion of prostaglandins by the conceptus. The aim of this study was to determine the expression and localization of key enzymes of the different pathways leading to synthesis of prostaglandin E2 and F2α in the equine conceptus during the mobility phase. Enzyme expression was analyzed via quantitative RT-PCR in total RNA samples of equine conceptuses collected on days 10 (n = 5), 12 (n = 12), 14 (n = 5) and 16 (n = 7) from healthy mares. Relative abundance of cyclooxygenase (COX)-2 mRNA was higher (p < 0.05) than of COX-1 irrespective of conceptus age and for phospholipase A2 on day 16 in comparison to all other days (p < 0.01). Abundance of mRNA of cytosolic and microsomal prostaglandin E synthase (PGES) and of carbonyl reductase (CBR) 1 was not influenced by conceptus age. Immunohistochemically, COX-1, COX-2, as well as cytosolic and microsomal PGES were present in both the ectodermal and endodermal layer of the yolk sac wall. CBR-1 was restricted to periembryonic disc area. The localisation of the key enzymes explains the mechanism of embryo mobility. In vitro incubation of primary trophoblast cell cultures with oxytocin had no effect on key enzyme synthesis. Full article
(This article belongs to the Special Issue Challenges in the Reproduction of Equids)
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Review

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13 pages, 2051 KiB  
Review
Antimicrobial Resistance in Equine Reproduction
by Pongpreecha Malaluang, Elin Wilén, Johanna Lindahl, Ingrid Hansson and Jane M. Morrell
Animals 2021, 11(11), 3035; https://doi.org/10.3390/ani11113035 - 22 Oct 2021
Cited by 19 | Viewed by 4658
Abstract
Bacteria develop resistance to antibiotics following low-level “background” exposure to antimicrobial agents as well as from exposure at therapeutic levels during treatment for bacterial infections. In this review, we look specifically at antimicrobial resistance (AMR) in the equine reproductive tract and its possible [...] Read more.
Bacteria develop resistance to antibiotics following low-level “background” exposure to antimicrobial agents as well as from exposure at therapeutic levels during treatment for bacterial infections. In this review, we look specifically at antimicrobial resistance (AMR) in the equine reproductive tract and its possible origin, focusing particularly on antibiotics in semen extenders used in preparing semen doses for artificial insemination. Our review of the literature indicated that AMR in the equine uterus and vagina were reported worldwide in the last 20 years, in locations as diverse as Europe, India, and the United States. Bacteria colonizing the mucosa of the reproductive tract are transferred to semen during collection; further contamination of the semen may occur during processing, despite strict attention to hygiene at critical control points. These bacteria compete with spermatozoa for nutrients in the semen extender, producing metabolic byproducts and toxins that have a detrimental effect on sperm quality. Potential pathogens such as Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa may occasionally cause fertility issues in inseminated mares. Antibiotics are added during semen processing, according to legislation, to impede the growth of these microorganisms but may have a detrimental effect on sperm quality, depending on the antimicrobial agent and concentration used. However, this addition of antibiotics is counter to current recommendations on the prudent use of antibiotics, which recommend that antibiotics should be used only for therapeutic purposes and after establishing bacterial sensitivity. There is some evidence of resistance among bacteria found in semen samples. Potential alternatives to the addition of antibiotics are considered, especially physical removal separation of spermatozoa from bacteria. Suggestions for further research with colloid centrifugation are provided. Full article
(This article belongs to the Special Issue Challenges in the Reproduction of Equids)
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