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Topic:Pregnancy

Pregnancy in horses, or equine gestation, is a physiological process that involves the development of a foal within the mare over approximately 11 months. This period is characterized by distinct stages, including fertilization, embryonic development, and fetal growth. Throughout gestation, mares undergo various physiological and hormonal changes to support the developing fetus. Monitoring pregnancy in horses involves assessing fetal health and mare well-being through veterinary examinations and diagnostic tools such as ultrasound. This page compiles peer-reviewed research studies and scholarly articles that explore the physiological processes, management practices, and health considerations associated with equine pregnancy.
Effect of Using Two Cryopreservation Methods on Viability and Fertility of Frozen Stallion Sperm.
Journal of equine veterinary science    October 24, 2018   Volume 72 37-40 doi: 10.1016/j.jevs.2018.10.008
Maziero RRD, Guaitolini CRF, Guasti PN, Monteiro GA, Martin I, Silva JPMD, Crespilho AM, Papa FO.Studies involving different methods and techniques of cryopreservation and its interactions with the conception rates in artificial insemination (AI) programs are reported in the literature. This study evaluated the sperm kinetics, plasma membrane integrity, and fertility rates of mares inseminated with cryopreserved stallion semen subjected to different freezing methods. For this, four ejaculates from five stallions were collected and frozen in conventional (Styrofoam box) or automated system in Mini-Digitcool ZH 400. Seminal samples were evaluated after thawing for sperm motion parameters by...
Factors affecting the likelihood of pregnancy and embryonic loss after transfer of cryopreserved in vitro produced equine embryos.
Equine veterinary journal    October 24, 2018   Volume 51, Issue 4 446-450 doi: 10.1111/evj.13028
Claes A, Cuervo-Arango J, van den Broek J, Galli C, Colleoni S, Lazzari G, Deelen C, Beitsma M, Stout TA.In vitro embryo production (IVEP) is increasingly popular but data assessing the outcome of transferred embryos are scarce. Objective: To determine the likelihood of pregnancy and embryonic loss after transfer of frozen-thawed IVP embryos and identify factors influencing success. Methods: Retrospective clinical study. Methods: Blastocysts (n = 261) were produced from immature oocytes of Warmblood mares (n = 116) by Intracytoplasmic Sperm Injection (ICSI) and in vitro culture, and cryopreserved. Thawed IVP embryos were transferred into recipient mares on day 4, 5 or 6 after ovulation. The ...
Multivariable analysis to determine risk factors associated with early pregnancy loss in thoroughbred broodmares.
Theriogenology    October 9, 2018   Volume 124 18-23 doi: 10.1016/j.theriogenology.2018.10.008
de Mestre AM, Rose BV, Chang YM, Wathes DC, Verheyen KLP.Early pregnancy loss (EPL) between days 15-65 after breeding has been shown to occur in 7.9% of equine pregnancies with substantial economical, welfare and safety implications. Whilst maternal age has been recognised as an important risk factor in relation to the incidence of EPL, few other risk factors have been conclusively identified. Further, multivariable data analysis of risk factors for EPL is sparse. A prospective cohort investigation of thoroughbred broodmares in the United Kingdom was conducted over the 2013 and 2014 breeding seasons. Information relating to 28 factors including mare...
Dynamic changes in gene expression and signalling during trophoblast development in the horse.
Reproduction (Cambridge, England)    October 1, 2018   Volume 156, Issue 4 313-330 doi: 10.1530/REP-18-0270
Read JE, Cabrera-Sharp V, Offord V, Mirczuk SM, Allen SP, Fowkes RC, de Mestre AM.Equine chorionic girdle trophoblast cells play important endocrine and immune functions critical in supporting pregnancy. Very little is known about the genes and pathways that regulate chorionic girdle trophoblast development. Our aim was to identify genes and signalling pathways active in vivo in equine chorionic girdle trophoblast within a critical 7-days window. We exploited the late implantation of the equine conceptus to obtain trophoblast tissue. An Agilent equine 44K microarray was performed using RNA extracted from chorionic girdle and chorion (control) from equine pregnancy days 27, ...
Dynamic changes in gene expression and signalling during trophoblast development in the horse.
Reproduction (Cambridge, England)    October 1, 2018   Volume 156, Issue 4 313–330 doi: 10.1530/REP-18-0270
Read JE, Cabrera-Sharp V, Offord V, Mirczuk SM, Allen SP, Fowkes RC, de Mestre AM.Equine chorionic girdle trophoblast cells play important endocrine and immune functions critical in supporting pregnancy. Very little is known about the genes and pathways that regulate chorionic girdle trophoblast development. Our aim was to identify genes and signalling pathways active in vivo in equine chorionic girdle trophoblast within a critical 7-days window. We exploited the late implantation of the equine conceptus to obtain trophoblast tissue. An Agilent equine 44K microarray was performed using RNA extracted from chorionic girdle and chorion (control) from equine pregnancy days 27, ...
Cell type-specific endometrial transcriptome changes during initial recognition of pregnancy in the mare.
Reproduction, fertility, and development    September 27, 2018   Volume 31, Issue 3 496-508 doi: 10.1071/RD18144
Scaravaggi I, Borel N, Romer R, Imboden I, Ulbrich SE, Zeng S, Bollwein H, Bauersachs S.Previous endometrial gene expression studies during the time of conceptus migration did not provide final conclusions on the mechanisms of maternal recognition of pregnancy (MRP) in the mare. This called for a cell type-specific endometrial gene expression analysis in response to embryo signals to improve the understanding of gene expression regulation in the context of MRP. Laser capture microdissection was used to collect luminal epithelium (LE), glandular epithelium and stroma from endometrial biopsies from Day 12 of pregnancy and Day 12 of the oestrous cycle. RNA sequencing (RNA-Seq) showe...
The effect of flunixin meglumine, firocoxib and meloxicam on the uterine mobility of equine embryos.
Theriogenology    September 25, 2018   Volume 123 132-138 doi: 10.1016/j.theriogenology.2018.09.026
Okada CTC, Andrade VP, Freitas-Dell'Aqua CP, Nichi M, Fernandes CB, Papa FO, Alvarenga MA.Embryo mobility occurs as a result of prostaglandin production by the embryo and endometrium, promoting uterine smooth muscle contractions, which propels the embryonic vesicle through the lumen. Non-steroidal anti-inflammatory drugs (NSAIDs), as flunixin meglumine, are routinely used in equine medicine and can alter the conceptus mobility if applied in early pregnancy, which may impair maternal recognition of pregnancy. The objective of this study was to evaluate and compare the effect of flunixin meglumine (FM; 1.1 mg/kg IV), firocoxib (FIRO; 0.2 mg/kg PO), and meloxicam (ML; 0.6 mg/kg,...
Ultrastructural and histological characteristics of the endometrium during early embryo development in mares.
Theriogenology    September 18, 2018   Volume 123 1-10 doi: 10.1016/j.theriogenology.2018.09.018
Camozzato GC, Martinez MN, Bastos HBA, Fiala-Rechsteiner S, Meikle A, Jobim MIM, Gregory RM, Mattos RC.The aim of this study was to evaluate ultrastructural and histological changes in the endometrium on days 7, 10 and 13 post-ovulation in pregnant and cyclic mares. Mares were routinely examined by transrectal palpation and ultrasonographic examination of the reproductive tract until estrus was detected. In the first cycle, endometrial biopsies from 30 cyclic mares (Cyclic group) were collected on days 7, 10 and 13 post-ovulation. In the second cycle, the same mares were bred by a fertile stallion. At days 7, 10 and 13 post-ovulation intrauterine biopsies were collected. Immediately after sampl...
Non-surgical embryo transfer technique and recipient mare pregnancy rate.
The Veterinary record    September 16, 2018   Volume 183, Issue 10 320-322 doi: 10.1136/vr.k3700
Card C.No abstract available
Assessment of equine alpha-fetoprotein levels in mares and newborn foals in the periparturient period.
Theriogenology    September 8, 2018   Volume 122 53-60 doi: 10.1016/j.theriogenology.2018.08.026
Vincze B, Solymosi N, Debnár V, Kútvölgyi G, Krikó E, Wölfling A, Szenci O.Alpha-fetoprotein (AFP) is best known in human obstetrics for its association with fetal anomalies recognized in the 1970s. Although this fetal protein had been shown to be present in the sera of many mammalian species, its possible diagnostic role in the detection of abnormalities was evaluated only later, when a research laboratory published variable levels of AFP in different groups of mares with pregnancy problems (twins, conception failure, placentitis, embryonic loss), and subsequently differences were demonstrated in its serum levels between aborted and healthy mares. In this study, per...
Changes in maternal pregnane concentrations in mares with experimentally-induced, ascending placentitis.
Theriogenology    September 7, 2018   Volume 122 130-136 doi: 10.1016/j.theriogenology.2018.09.001
Wynn MAA, Ball BA, May J, Esteller-Vico A, Canisso I, Squires E, Troedsson M.The objectives of this study were to compare via liquid chromatography-tandem mass spectrometry (LC-MS/MS) progesterone (P4), 5α-dihydroprogesterone (DHP), allopregnanolone, 3β-hydroxy-5α-pregnan-20-one (3β5P), 20α-hydroxy-5α-pregnan-3-one (20α5P), 5α-pregnan-3β,20α-diol (βα-diol), and 5α-pregnan-3β,20β-diol (ββ-diol) concentrations in plasma of mares with experimentally-induced, ascending placentitis compared to gestationally age-matched control mares. Placentitis was induced via intracervical inoculation of Streptococcus equi spp. zooepidemicus between 260 and 280 days of ge...
Advanced mare age impairs the ability of in vitro-matured oocytes to correctly align chromosomes on the metaphase plate.
Equine veterinary journal    August 9, 2018   Volume 51, Issue 2 252-257 doi: 10.1111/evj.12995
Rizzo M, Ducheyne KD, Deelen C, Beitsma M, Cristarella S, Quartuccio M, Stout TAE, de Ruijter-Villani M.Advanced mare age is associated with declining fertility and an increased risk of early pregnancy loss. Compromised oocyte quality is probably the primary reason for reduced fertility, but the defects predisposing to embryonic death are unknown. In women, advanced age predisposes to chromosome segregation errors during meiosis, which lead to embryonic aneuploidy and a heightened risk of miscarriage. Objective: To evaluate the effect of advanced mare age on chromosome alignment and meiotic spindle morphology in in vitro-matured (IVM) oocytes. Methods: Morphometric and morphological analysis. M...
Intracellular calcium chelating agent (BAPTA-AM) aids stallion semen cooling and freezing-thawing.
Reproduction in domestic animals = Zuchthygiene    July 9, 2018   Volume 53, Issue 5 1235-1242 doi: 10.1111/rda.13245
Wu S, Canisso IF, Yang W, Ul Haq I, Liu Q, Han Y, Zeng S.This study aimed to investigate the effects of different concentrations of 1,2-bis-(o-aminophenoxy)-ethane-N,N,N0 N0-tetraacetic acid, tetra-acetoxymethyl ester (BAPTA-AM), an intracellular calcium chelating agent, on stallion semen cooling and freezing-thawing. After collection, semen was extended (1:1 v/v) on a skim milk-based extender, centrifuged and resuspended at 400 million/ml into cooling or freezing extenders containing 0, 5, 25, 50, 100 and 200 μΜ BAPTA-AM. Motility parameters were assessed after cooling in Equitainer at 5°C for 12, 24, 48, 72 and 120 hr and after freezing-tha...
Expression of activin receptors in the equine uteroplacental tissue: an immunohistochemical analysis.
Journal of equine science    July 6, 2018   Volume 29, Issue 2 33-37 doi: 10.1294/jes.29.33
Kimura Y, Sasaki M, Watanabe K, Dhakal P, Sato F, Taya K, Nambo Y.Activin is secreted from equine uterine glands and plays important roles in establishment and maintenance of pregnancy in mares. This study aimed to localize activin receptors (ActRs) IA/B and IIA/B using immunohistochemistry in the uteroplacental tissues of seven pregnant Thoroughbred mares. At the time of tissue collection, the mares were at the following days of pregnancy: 88, 120, 161, 269, 290, 313, and 335 days. We fixed the uteroplacental tissues in 4% paraformaldehyde and obtained serial sections that were subsequently stained for analysis. All four isoforms of ActR were expressed in t...
Association of equine sperm population parameters with outcome of intracytoplasmic sperm injections.
Theriogenology    July 5, 2018   Volume 119 114-120 doi: 10.1016/j.theriogenology.2018.06.027
Gonzalez-Castro RA, Carnevale EM.Limited clinical information is available regarding sperm population parameters that are important for use with equine intracytoplasmic sperm injection (ICSI). Therefore, the appropriateness of a sample of sperm is typically not known before ICSI. The aim of our study was to determine which sperm population characteristics were predictive of ICSI outcome. Frozen-thawed sperm samples (n = 114) from 37 stallions in a clinical program were analyzed after ICSI for percentages of normal morphology (MORPH+), live as assessed by eosin/nigrosin stain (LIVE+), membrane intact as assessed by hypoosmot...
Expression of glucose transporters in the endometrium and early conceptus membranes of the horse.
Placenta    June 25, 2018   Volume 68 23-32 doi: 10.1016/j.placenta.2018.06.308
Gibson C, de Ruijter-Villani M, Rietveld J, Stout TAE.Glucose is the primary energy substrate for early conceptus development and, for the first 40 days of gestation, the equine conceptus depends solely on glucose available in the histotroph; thereafter, histotrophic glucose provision continues to support transport across the definitive placenta. To investigate glucose provision routes during early equine pregnancy we examined expression of glucose transporters in conceptus membranes and endometrium recovered on days 7, 14, 21 and 28 after ovulation. To further differentiate the contributions of maternal progesterone priming and conceptus-endomet...
Electrocardiographic confirmation of a twin pregnancy in a mare at 8 months of gestation.
Journal of veterinary cardiology : the official journal of the European Society of Veterinary Cardiology    June 23, 2018   Volume 20, Issue 4 294-299 doi: 10.1016/j.jvc.2018.05.003
Vera L, Decloedt A, Van Steenkiste G, De Clercq D, Govaere J, van Loon G.A 15-year-old Warmblood mare, at 8 months of gestation, was presented to the Ghent University Faculty of Veterinary Medicine for evaluation of suspected stage one labor. Transrectal ultrasonography revealed no clear signs of placentitis or stage one labor. The combined thickness of uterus and placenta was 11 mm and the cervix was closed. Twin pregnancy was suspected by transabdominal ultrasonography but could not be confirmed with certainty. An electrocardiogram (ECG) was recorded from the mare's abdominal wall to register a fetal ECG. To confirm twin pregnancy, registration of both fetal ECG...
Amino acid transporter expression in the endometrium and conceptus membranes during early equine pregnancy.
Reproduction, fertility, and development    June 16, 2018   Volume 30, Issue 12 1675-1688 doi: 10.1071/RD17352
Gibson C, de Ruijter-Villani M, Rietveld J, Stout TAE.Maternally derived amino acids (AA) are essential for early conceptus development, and specific transporters enhance histotrophic AA content during early ruminant pregnancy. In the present study we investigated AA transporter expression in early equine conceptuses and endometrium, during normal pregnancy and after induction of embryo-uterus asynchrony. 'Normal' conceptuses and endometrium were recovered on Days 7, 14, 21 and 28 after ovulation. To investigate asynchrony, Day 8 embryos were transferred to recipient mares on Day 8 or Day 3, and conceptuses were recovered 6 or 11 days later. Endo...
[Clinical signs in late pregnant mares].
Tierarztliche Praxis. Ausgabe G, Grosstiere/Nutztiere    June 14, 2018   Volume 46, Issue 3 164-171 doi: 10.15653/TPG-180125
Neuhauser S, Gösele P, Handler J.During the peripartal period, interpretation of basic clinical signs may be challenging. In the present study, heart rate (HR), respiratory rate (RR) and body temperature (BT) were evaluated in healthy mares of different breed types and compared to reference values for adult horses from the literature. Methods: During daily physical exams of periparturient mares, the HR, RR and BT were evaluated. Differences according to the horse's size were investigated and in large breeds, the influence of dystocia or retained placenta was analysed. Results: During the last weeks before parturition (a. p.),...
Effects of mares’ age and day of gestation on efficacy of transvaginal ultrasound-guided twin reduction.
Tierarztliche Praxis. Ausgabe G, Grosstiere/Nutztiere    June 14, 2018   Volume 46, Issue 3 172-177 doi: 10.15653/TPG-170741
Rau J, Tiedemann D, Sielhorst J, Tönissen A, Burger D, Martinsson G, Rohn K, Oldenhof H, Sieme H.Transvaginal ultrasound-guided aspiration (TUA) is a procedure which can be used for the reduction of twins post-fixation in the mare. The aim of this study was to evaluate the effect of the age of mares and the day of gestation on the outcome of TUA treatment. Methods: In 88 mares, diagnosed pregnant with twins, TUA of the yolk sac or allantoic fluid was performed between day 30 and 62 of gestation. Mares were aged 3-22 years. Ultrasonographic examination for a viable singleton pregnancy was performed by referring veterinarians 5-7 days and 4 weeks after TUA. Based on reported findings, effec...
Equine foetal gender determination in mid- to late gestational mares: A practical inquiry.
Reproduction in domestic animals = Zuchthygiene    June 12, 2018   Volume 53, Issue 5 1027-1032 doi: 10.1111/rda.13211
Van de Velde M, Roels K, Ververs C, Gerits I, Govaere J.In recent years, the interest in equine foetal gender determination (FGD) during gestation increased remarkably. Ultrasonographic FGD can be performed in two different periods during gestation. The earliest examination can take place at a gestational age of 60-70 days, whereby the genital tubercle is used to differentiate between male and female foeti. The time window of the second approach is wider (120-210 days), and there are more characteristics to take into consideration. In this article, the feasibility and accuracy of ultrasonographic FGD in mid- to late gestation are evaluated. One h...
5α-dihydroprogesterone concentrations and synthesis in non-pregnant mares.
The Journal of endocrinology    June 2, 2018   Volume 238, Issue 1 25-32 doi: 10.1530/JOE-18-0215
Conley AJ, Scholtz EL, Legacki EL, Corbin CJ, Knych HK, Dujovne GD, Ball BA, Moeller BC, Stanley SD. and evidence indicates that the bioactive, 5α-reduced progesterone metabolite, 5α-dihydroprogesterone (DHP) is synthesized in the placenta, supporting equine pregnancy, but its appearance in early pregnancy argues for other sites of synthesis also. It remains unknown if DHP circulates at relevant concentrations in cyclic mares and, if so, does synthesis involve the non-pregnant uterus? Jugular blood was drawn daily from cyclic mares ( = 5). Additionally, ovariectomized mares (OVX) and geldings were administered progesterone (300 mg) intramuscularly. Blood was drawn before and after t...
Influences of intrauterine semen administration on regulatory T lymphocytes in the oestrous mare (Equus caballus).
Theriogenology    June 1, 2018   Volume 118 119-125 doi: 10.1016/j.theriogenology.2018.05.030
Hartmann C, Gerner W, Walter I, Saalmüller A, Aurich C.In the mare, early pregnancy loss is common, but involvement of the maternal immune system in the pathogenesis of this condition has not been investigated in detail so far. In the present study, we assessed effects of exposure of the endometrium to semen or seminal plasma in oestrous mares on the response of regulatory T lymphocytes (Tregs) in the peripheral circulation as well as in the endometrium. Raw semen, seminal plasma or PBS (control) were introduced into the uterus of oestrous mares (n = 12). Blood was collected immediately before insemination or PBS infusion (time 0), and 12, 24 ...
Temporal relationships of GH, IGF-I and fructosamine concentrations in pregnant Spanish Purebred mares: A substantial contribution from the hormonal standpoint.
Theriogenology    May 30, 2018   Volume 118 164-171 doi: 10.1016/j.theriogenology.2018.05.040
Satué K, Marcilla M, Medica P, Cravana C, Fazio E.Growth hormone (GH) and insulin-like growth factor (IGF-I) exert essential functions to maintain an adequate metabolic activity for correct placental and fetal development. Although fructosamine (FRUCT) is used in clinical evaluation to assess the glycaemic state, the relationships between GH, IGF-I and FRUCT remain unknown in the mare. Therefore, the objectives of this study were to evaluate the relationships among these parameters in pregnant mares. A total of 30 healthy Spanish Purebred mares, with an age range of 9.33 ± 3.31 years, were studied during the 11 months of gestation. Serum...
The development of in vitro embryo production in the horse.
Equine veterinary journal    May 25, 2018   Volume 50, Issue 6 712-720 doi: 10.1111/evj.12839
Morris LHA.The development of techniques to produce equine embryos in vitro is reviewed with specific reference to intracytoplasmic sperm injection (ICSI). Unexplored 50 years ago, this technology has progressed rapidly in the last 20 years to become a commercial reality for the equine breeding industry. Improvements in our understanding of oocyte and embryo competence in the horse have been key factors in overcoming some of the initial problems associated with ICSI. It is now possible to obtain high nuclear maturation and cleavage rates in vitro and the most limiting factor, presently, is the low rate o...
Effect of embryo transfer technique on the likelihood of pregnancy in the mare: a comparison of conventional and Wilsher’s forceps-assisted transfer.
The Veterinary record    May 24, 2018   Volume 183, Issue 10 323 doi: 10.1136/vr.104808
Cuervo-Arango J, Claes AN, Stout TA.The aim of this study was to compare the success of embryo transfer (ET) performed by veterinarians with different degrees of experience using one of two ET techniques. Over three years, 179 embryos were transferred by three operators with moderate to high experience using a 'conventional' manual technique, and 170 embryos were transferred by four operators with little or no previous ET experience using a 'Wilsher' technique (Polansky speculum and Wilsher's cervical forceps). The pregnancy status of recipient mares at the first pregnancy diagnosis and the embryo loss by the last examination we...
Evaluation of environmental effects on reproductive characteristics of Mangalarga Marchador mares in a commercial embryo transfer program.
Animal reproduction science    May 15, 2018   Volume 195 131-138 doi: 10.1016/j.anireprosci.2018.05.016
Rua MAS, Quirino CR, Rodrigues ACC, Christo MM, Barreto MAP.The objective of this study was to evaluate the environmental effects on embryo recovery rate and pregnancy rate of Mangalarga Marchador mares. The reproductive characteristics of donor and recipient mares were evaluated during five years in Brazilian tropical environment. The mares were used throughout the year and seasons were classified as: October to April (breeding season - BS); May (autumn transition out of the breeding season - ATBS); June to August (non-breeding season - nBS); and September (vernal transition into the breeding season - VTBS). Daily temperature rainfall and hours of day...
Interferon epsilon is constitutively expressed in equine endometrium and up-regulated during the luteal phase.
Animal reproduction science    May 12, 2018   Volume 195 38-43 doi: 10.1016/j.anireprosci.2018.05.003
Fischer CD, Wachoski-Dark GL, Grant DM, Bramer SA, Klein C.Interferon epsilon (IFNE) is type I interferon which stands out through its unusual expression profile and differing regulation compared to classic type I interferons such as interferon alpha and interferon beta. Unlike other type I interferons, the expression of IFNE is not stimulated through exposure to viral agents. Expression of IFNE is most abundant in mouse and human endometrium where it is constitutively expressed in luminal and glandular epithelial cells and expression levels are up-regulated with estrogen exposure. The aim of the current study was to determine whether a cycle or pregn...
Sequential concentrations of placental growth factor and haptoglobin, and their relation to oestrone sulphate and progesterone in pregnant Spanish Purebred mare.
Theriogenology    April 27, 2018   Volume 115 77-83 doi: 10.1016/j.theriogenology.2018.04.033
Satué K, Marcilla M, Medica P, Ferlazzo A, Fazio E.The objectives of this study were to establish reference values for serum concentrations of placental growth factor (PlGF) and haptoglobin (Hp), and to analyze whether the levels of oestrone sulphate (E1S) and progesterone (P4) are physiologically involved in the dynamic modifications of the above parameters in pregnant mares. A total of 30 healthy Spanish Purebred mares ranging in age 9.33 ± 3.31 years were studied during the 11 months of gestation. Serum concentrations of PlGF were detected by EIA, Hp using commercial Phase Haptoglobin assay and E1S and P4 levels through RIA. The serum ...
Horse embryo diameter is influenced by the embryonic age but not by the type of semen used to inseminate donor mares.
Theriogenology    April 27, 2018   Volume 115 90-93 doi: 10.1016/j.theriogenology.2018.04.023
Cuervo-Arango J, Claes AN, Stout TAE.The diameter of embryos recovered from mares on Day 8 after ovulation varies greatly, from as little as 130 μm to as much as 2500 μm. Several factors have been proposed to affect embryo size at recovery, one of which is the type of semen (frozen vs fresh or cooled-transported) used to inseminate the mare. In addition, it has been shown that smaller embryos (<300 μm) recovered on Day 8 are less likely than larger embryos to result in successful pregnancy after transfer. However, whether the actual age of the embryo (interval from fertilization to flushing) in relation to its size also ...
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