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Topic:Reproductive Technology

Reproductive technology in horses encompasses a range of scientific techniques and procedures aimed at assisting and enhancing equine reproduction. These technologies include artificial insemination, embryo transfer, and cryopreservation of gametes and embryos. They are employed to improve breeding efficiency, manage genetic diversity, and preserve valuable genetic material. Artificial insemination involves the collection and introduction of semen into the mare's reproductive tract, while embryo transfer allows for the harvesting and implantation of embryos from donor to recipient mares. Cryopreservation involves freezing and storing sperm, oocytes, or embryos for future use. This page compiles peer-reviewed research studies and scholarly articles that investigate the methodologies, applications, and outcomes of reproductive technologies in equine breeding and management.
Cryopreservation and fertility of ejaculated and epididymal stallion sperm.
Animal reproduction science    August 16, 2011   Volume 127, Issue 3-4 197-201 doi: 10.1016/j.anireprosci.2011.08.002
Monteiro GA, Papa FO, Zahn FS, Dellaqua JA, Melo CM, Maziero RR, Avanzi BR, Alvarenga MA, Guasti PN.The cryopreservation of epididymal sperm is important to preserve genetic material from valuable deceased males. This study evaluated the viability of sperm samples from eight stallions under three conditions: (1) collected using an artificial vagina (EJ-0h), (2) recovered from the epididymal cauda immediately after orchiectomy (EP-0h), and (3) recovered from the epididymal cauda after 24h of storage at 5°C (EP-24h). To obtain EJ-0h sperm, two ejaculates were collected from each stallion. After 1 week, the stallions were submitted to bilateral orchiectomy, and one of the removed epididymides ...
Cryopreservation of immature equine oocytes, comparing a solid surface vitrification process with open pulled straws and the use of a synthetic ice blocker.
Theriogenology    August 10, 2011   Volume 77, Issue 1 21-27 doi: 10.1016/j.theriogenology.2011.07.008
de Leon PM, Campos VF, Corcini CD, Santos EC, Rambo G, Lucia T, Deschamps JC, Collares T.The objective was to evaluate the effect of three cryopreservation methods on the in vitro maturation (IVM) and membrane integrity (MIn) of immature equine oocytes. An open pulled straw (OPS) method, a novel solid surface vitrification (SSV) process, and the addition of a synthetic ice blocker were evaluated. Compared with the control group (N=269), the OPS (N=159) and the SSV (N=202) cryopreservation methods decreased both IVM (50.9 vs. 13.3 and 9.4%, respectively; P<0.001) and MIn (76.6 vs. 31.1 and 33.7%; P<0.001) of immature equine oocytes. However, inclusion of 0.1% ice blocker in t...
Processing stored stallion semen doses by Single Layer Centrifugation.
Theriogenology    July 28, 2011   Volume 76, Issue 8 1424-1432 doi: 10.1016/j.theriogenology.2011.06.011
Morrell JM, Garcia BM, Pena FJ, Johannisson A.The purpose of this study was to determine if the quality of stored stallion semen doses could be enhanced by the scaled-up version of Single Layer Centrifugation using Androcoll-E-Large. Three semen doses from each of fifteen stallions were transported overnight to the Swedish University of Agricultural Sciences (SLU) for processing 24 h after semen collection. Sperm quality in the resulting SLC-selected samples was significantly improved compared to the uncentrifuged samples: mean progressive motility was increased by 8% on the day of processing (P < 0.001) and by 13% after 24 h cold stor...
Freezing stallion semen with the new Cáceres extender improves post thaw sperm quality and diminishes stallion-to-stallion variability.
Animal reproduction science    July 23, 2011   Volume 127, Issue 1-2 78-83 doi: 10.1016/j.anireprosci.2011.07.009
Rodríguez AM, Ferrusola CO, García BM, Morrell JM, Martínez HR, Tapia JA, Peña FJ.Ejaculates from 7 stallions were split and simultaneously frozen in three different extenders, INRA 96 egg yolk glycerol, Ghent and the newly developed extender Caceres. After thawing, samples were evaluated for motility (CASA system) sperm membrane integrity and early membrane changes (YoPro-1/Eth staining), acrosome integrity (FICT-PNA), and mitochondrial membrane potential (JC-1) (flow cytometry). Samples frozen in Caceres extender consistently showed the best results in post-thaw motility (increases ranging from 11 to 17%, p<0.05) and velocity (p<0.05), membrane integrity (increases ...
In vitro maturation and artificial activation of donkey oocytes.
Theriogenology    May 23, 2011   Volume 76, Issue 4 700-704 doi: 10.1016/j.theriogenology.2010.10.039
Zhao G, Wu K, Cui L, Zhao L, Liu Y, Tan X, Zhou H.Three media were evaluated for their ability to support in vitro maturation of donkey (Equus asinus) oocytes and their development after parthenogenetic activation. The basal medium for Medium 1 (M1) and Medium 2 (M2) was M199 and DMEM/F12 respectively, whereas, Medium 3 (M3) consisted of equal parts (v/v) of M199 and DMEM/F12. All three media were supplemented with 10% (v/v) fetal calf serum, 0.01 units/mL porcine FSH, 0.01 units/mL equine LH, 200 ng/mL insulin-like growth factor 1(IGF-I), 10 μl/mL insulin-transferrin-selenium (ITS), 0.1 mg/mL taurine, 0.1 mg/mL L-cysteine, 0.05 mg/mL L-glut...
Seminal parameters and field fertility of cryopreserved donkey jack semen after insemination of horse mares.
Equine veterinary journal    May 20, 2011   Volume 43, Issue 2 179-183 doi: 10.1111/j.2042-3306.2010.00130.x
Canisso IF, Carvalho GR, Morel MD, Ker PG, Rodrigues AL, Silva EC, Coutinho Da Silva MA.As mule production is often concentrated in remote areas of the world, a simplified semen cryopreservation protocol is required. Objective: To compare the seminal parameters of cryopreserved donkey semen in lactose-EDTA and lactose-yolk extenders and the fertility rates on horse mares. Methods: TRIAL 1: Sperm total and progressive motility, vigour (scale 0-5), morphology (major and minor defects) and plasma membrane integrity (HOST) were evaluated in 25 ejaculates from 5 donkey jacks immediately after collection (raw), after chilling to 5°C (chilled) and after freezing/thawing. The semen was ...
Density gradient centrifugation of sperm from a subfertile stallion and effect of seminal plasma addition on fertility.
Animal reproduction science    May 4, 2011   Volume 126, Issue 1-2 96-100 doi: 10.1016/j.anireprosci.2011.04.020
Mari G, Castagnetti C, Rizzato G, Mislei B, Iacono E, Merlo B.Stallions are not selected for fertility but for other criteria (pedigree, conformation, performances, progeny), therefore valuable but subfertile stallions with poor semen quality are frequently used in commercial breeding programs. The object of this study was to evaluate whether sperm selection through a silane-coated silica colloid gradient centrifugation, with or without the addition of seminal plasma of a high fertile stallion, could improve the pregnancy rates of an oligospermic valuable stallion in a commercial breeding program. In 2008 breeding season (experiment 1, n=104 mares), simp...
Follicle and systemic hormone interrelationships during induction of luteinized unruptured follicles with a prostaglandin inhibitor in mares.
Theriogenology    April 14, 2011   Volume 76, Issue 2 361-373 doi: 10.1016/j.theriogenology.2011.02.015
Cuervo-Arango J, Beg MA, Ginther OJ.The objective was to determine differences in follicle and reproductive hormone characteristics in mares with ovulatory and flunixin meglumine (FM)-induced anovulatory cycles. Estrous mares were given 1500 IU hCG when the follicle was ≥ 32 mm (0 h). In Experiment 1, control mares (n = 7) were not treated further. The remaining mares (n = 11) were given 1.7 mg/kg FM i.v. twice daily, from 0 to 36 h after hCG treatment. Blood samples and ultrasonographic examinations were performed every 12 h. All control mares ovulated normally between 36 and 48 h. In contrast, eight of 11 FM mares did not ov...
Effects of timing of induced luteolysis in embryo donor mares on reproductive performance and pregnancy rate in recipient mares.
Theriogenology    February 4, 2011   Volume 75, Issue 6 1170-1174 doi: 10.1016/j.theriogenology.2010.12.008
Goretti RG, Araújo RR, Filho AN, Araújo GH, Lopes EP, Guimarães JD.The objective was to evaluate the effects of giving prostaglandin F₂(α) (PGF) to donor mares 48 h prior to embryo collection. Non-lactating donor mares (n = 20 estrous cycles in 10 mares), ranging from 2.5 to 10 y of age and 400 to 500 kg of body weight were used from September 2004 to February 2005 in the southern hemisphere (Brazil). Donor mares were randomly assigned in a cross-over design study. During a Treated cycle, 7.5 mg PGF was given 48 h prior to embryo collection, whereas in the Control cycle, 7.5 mg PGF was given at embryo collection. In Treated Cycles, serum progesterone conce...
Quality and fertility of cooled-shipped stallion semen at the time of insemination.
Theriogenology    December 31, 2010   Volume 75, Issue 5 849-856 doi: 10.1016/j.theriogenology.2010.10.027
Heckenbichler S, Deichsel K, Peters P, Aurich C.Stallion semen processing is far from standardized and differs substantially between AI centers. Suboptimal pregnancy rates in equine AI may primarily result from breeding with low quality semen not adequately processed for shipment. It was the aim of the study to evaluate quality and fertility of cooled-shipped equine semen provided for breeding of client mares by commercial semen collection centers in Europe. Cooled shipped semen (n = 201 doses) from 67 stallions and 36 different EU-approved semen collection centers was evaluated. At arrival, semen temperature was 9.8 ± 0.2 °C, mean sperm ...
The effect of time of insemination with fresh cooled transported semen and natural mating relative to ovulation on pregnancy and embryo loss rates in the mare.
Reproduction in domestic animals = Zuchthygiene    November 30, 2010   Volume 46, Issue 4 678-681 doi: 10.1111/j.1439-0531.2010.01728.x
Newcombe JR, Cuervo-Arango J.One hundred and fifty-four mares were inseminated with fresh semen either during the pre- or post-ovulatory periods at different intervals relative to ovulation: 36-24 h (n = 17) and 24-0 h (n = 30) before ovulation; 0-8 h (n = 21), 8-16 h (n = 24), 16-24 h (n = 48) and 24-32 h (n = 14) h after ovulation. All mares received the same routine post-mating treatment consisting of an intrauterine infusion with 1 litre of saline and antibiotics followed 8 h later by an intravenous administration of oxytocin. Artificial inseminations (AI) from 36 h before ovulation up to 16 h post-ovulation were perf...
Pregnancies following artificial insemination with spermatozoa from problem stallion ejaculates processed by single layer centrifugation with Androcoll-E.
Reproduction in domestic animals = Zuchthygiene    November 30, 2010   Volume 46, Issue 4 642-645 doi: 10.1111/j.1439-0531.2010.01721.x
Morrell JM, Mari G, Kútvölgyi G, Meurling S, Mislei B, Iacono E, Rodriguez-Martinez H.Some stallions produce ejaculates of low quality and/or low fertility when used for artificial insemination (AI). The purpose of these five case studies was to use Single Layer Centrifugation (SLC) to select the best spermatozoa from 'problem' ejaculates for subsequent use in AI. Sperm quality, in terms of motility, morphology and chromatin integrity, was improved in the SLC-selected samples compared to the corresponding uncentrifuged samples, with the exception of one stallion thought to have ampullary stasis. In this stallion, neither the incidence of spermatozoa with detached heads nor the ...
Reducing the examination interval to detect ovulation below 12h does not improve pregnancy rates after postovulatory insemination with frozen/thawed semen in mares.
Animal reproduction science    November 12, 2010   Volume 123, Issue 1-2 60-63 doi: 10.1016/j.anireprosci.2010.11.004
Newcombe JR, Paccamonti D, Cuervo-Arango J.Data were analysed retrospectively from fourteen breeding seasons at an Equine Fertility Clinic for the effect of interval between pre- and postovulatory examinations for immediate postovulatory insemination on pregnancy rate (PR) and embryo loss rate (ELR). Mares of various breeds and ages were examined at intervals which varied from 0.5 to 15h between the pre- and postovulatory period over 867 cycles. When ovulation was detected they were inseminated with a single dose of commercial frozen-thawed semen. All mares were treated in the post-insemination period with intrauterine antibiotics and ...
Efficacy of tuohy needle in oocytes collection from excised mare ovaries.
Veterinary medicine international    August 5, 2010   Volume 2010 102591 doi: 10.4061/2010/102591
Cremonesi F, Anderson K, Lange-Consiglio A.TWO METHODS HAVE BEEN DESCRIBED TO RECOVER OOCYTES FROM EQUINE FOLLICLES IN EXCISED OVARIES: aspiration and scraping. Aim of this work was to develop an effective method for collecting equine oocytes using Tuohy needle and comparing this technique to aspiration and scraping, with or without tunica albuginea removal. This hollow hypodermic needle, usually employed for inserting epidural catheters, is designed with a slightly curved tip, shaped similar to a small curette. In unpeeled ovaries, the recovery rates of Tuohy needle group was higher (P < .05) than in the 16 g needle aspiration and ...
Effects of repeated embryo collection on embryo recovery rate in fertile mares.
Reproduction in domestic animals = Zuchthygiene    August 4, 2010   Volume 46, Issue 3 419-422 doi: 10.1111/j.1439-0531.2010.01682.x
Aurich C, König N, Budik S.In mares, repeated embryo collection in successive oestrous cycles is necessary if a greater number of foals should be produced. We investigated effects of repeated embryo collection in fertile donor mares on embryo recovery rates. In addition, an influence of the individual mare and season on embryo recovery rates was studied. In nine mares, a total of 153 embryo collections were performed during 30 months (17 ± 2.2 embryo collections per mare). The overall embryo recovery rate was 64% and did not differ among mares. Between successive embryo collection procedures, recovery rate varied signi...
Techniques for the insemination of low doses of stallion sperm.
Reproduction in domestic animals = Zuchthygiene    July 16, 2010   Volume 45 Suppl 2 35-39 doi: 10.1111/j.1439-0531.2010.01632.x
Samper JC, Plough T.In the last decade, there has been a significant increase in the quality and commercial use of frozen equine semen. The emergence of new reproductive technologies, coupled with the high prices for an insemination dose from some stallions, the increasing costs of import and export and the marketing policies of stallion agents or owners in the sport horse industry has stimulated the fractionation of doses for insemination. Consequently, the sperm number and the volume of an insemination dose are significantly reduced. To deliver lower doses of sperm in lower volumes compared to the standard dose...
Total RNA isolation from stallion sperm and testis biopsies.
Theriogenology    July 7, 2010   Volume 74, Issue 6 1099-1106e11062 doi: 10.1016/j.theriogenology.2010.04.023
Das PJ, Paria N, Gustafson-Seabury A, Vishnoi M, Chaki SP, Love CC, Varner DD, Chowdhary BP, Raudsepp T.Sperm mRNA transcriptional profiles can be used to evaluate male fertility, yet differences between species in sperm attributes and packaging require adjustments in sperm RNA isolation protocols. The objective was to optimize RNA isolation methodology for fresh, frozen, and extended ejaculates, and epididymal sperm of stallions. Additionally, a protocol for RNA isolation from testis biopsies was established. Separation of sperm from somatic cells was critical for assuring the isolation of sperm-specific RNA. The highest purity was obtained by centrifuging ejaculates and epididymal sperm at 200...
Factors affecting pre-ovulatory follicle diameter in the mare: the effect of mare age, season and presence of other ovulatory follicles (multiple ovulation).
Theriogenology    July 7, 2010   Volume 74, Issue 7 1241-1247 doi: 10.1016/j.theriogenology.2010.05.027
Morel MC, Newcombe JR, Hayward K.The importance of elucidating factors affecting reproductive performance and efficiency is of paramount concern to the equine industry. Oocyte viability is known to be one of the determinants of reproductive success and evidence suggests that it may be linked to follicle size. The aims of this study were, therefore, to ascertain: i) the average diameter and range of pre-ovulatory follicles in Thoroughbred mares; ii) whether this is affected by either mare age, time within the breeding season, or the presence of multiple pre-ovulatory follicles (MO). One thousand, four hundred and ninety two Th...
Reproductive parameters of Mangalarga Marchador mares in a commercial embryo transfer programme.
Reproduction in domestic animals = Zuchthygiene    June 23, 2010   Volume 46, Issue 2 261-267 doi: 10.1111/j.1439-0531.2010.01656.x
Lopes Ede P, Siqueira JB, Pinho RO, Guimarães JD, Rocha AN, de Carvalho GR, Torres CA.The objective of this study is to evaluate the reproductive efficiency in donors and recipient Mangalarga Marchador mares in commercial programmes of embryo transfer (ET) and the effects of some reproductive characteristics and ET methodology on conception rates in the recipient mares. A total of 1140 flushing procedures were performed and 830 embryos (72.8%) were recovered. There were no differences between the rates of embryonic recovery in the different breeding seasons (p > 0.05) and 92.8% of the recovered embryos were 8-9 days old. There was no difference in the embryonic recovery rega...
Reproductive efficiency of Thoroughbred and Standardbred horses in north-east Victoria.
Australian veterinary journal    June 10, 2010   Volume 88, Issue 5 169-175 doi: 10.1111/j.1751-0813.2010.00565.x
Nath LC, Anderson GA, McKinnon AO.To evaluate the reproductive efficiency of horse farms in north-east Victoria and identify aspects of management to be targeted for improving reproductive efficiency. Methods: Retrospective study. Methods: Records from seven Thoroughbred (TB) and four Standardbred (STB) studs in north-east Victoria from 1990 to 2001 were reviewed; 8813 cycles in 4455 mares were analysed. TB mares were inseminated by natural mating, whereas STB mares (89%) were artificially inseminated. Results: The overall early pregnancy rate per cycle was 68.8% for TB mares and for STB mares, 68.3%. Multiple pregnancy per cy...
Characterizing the meiotic spindle configuration and chromosome complement of in vivo matured equine oocytes.
Animal reproduction science    May 20, 2010   Volume 121, Issue 1-2 Suppl 234-236 doi: 10.1016/j.anireprosci.2010.04.136
Vanderwall DK, Baumann C, Viveiros M, Sertich PL, Kelleman AA, Maenhoudt C, Jacobson CC, De La Fuente R.No abstract available
Enhancing reproductive performance in mares.
Compendium (Yardley, PA)    May 18, 2010   Volume 32, Issue 1 E1-E7 
Scherzer J.Reproductive performance in mares can be enhanced by various techniques. Protocols hastening the onset of follicular development help establish pregnancy in mares and ensure that foals are born early in the year. The time spent breeding mares can be reduced by synchronizing estrus and inducing ovulation. After successful fertilization of the oocyte, the developing embryo can survive in the uterus only if postbreeding endometritis, if present, is treated.
Sex, science and satisfaction: a heady brew.
Animal reproduction science    April 21, 2010   Volume 121, Issue 1-2S S262-S278 doi: 10.1016/j.anireprosci.2010.04.178
Allen WR.Genus-specific reproductive processes and strategies displayed by equids, camelids and elephantids are compared and contrasted to illustrate the amazing diversity of reproductive physiology between genera and the equally surprising conservation of reproductive processes across a genus in the face of other dramatic phenotypic modifications and adaptation to prosper in the prevailing environment. From intrauterine conceptus mobility and chorionic gonadotrophic secretion by specialised invasive trophoblast cells in equids, through induced ovulation, an asymmetrical uterus and an almost absurdly s...
Single layer centrifugation of stallion spermatozoa improves sperm quality compared with sperm washing.
Reproductive biomedicine online    April 9, 2010   Volume 21, Issue 3 429-436 doi: 10.1016/j.rbmo.2010.03.027
Morrell JM, Rodriguez-Martinez H, Johannisson A.This study compared the effect on semen quality of different handling methods used in the preparation of stallion semen doses for artificial insemination. The three methods were (i) extending the ejaculate to 30-50 x 10(6)/ml, (ii) single layer centrifugation (SLC) and (iii) sperm washing (centrifugation without a colloid). An additional treatment was to add seminal plasma (SP) in various proportions to some SLC preparations. The resulting samples were evaluated for sperm motility by computer assisted sperm analysis, membrane integrity using the Nucleocounter SP-100 and chromatin integrity by ...
Addition of ficoll and disaccharides to vitrification solutions improve in vitro viability of vitrified equine embryo.
Cryo letters    March 24, 2010   Volume 30, Issue 6 408-413 
Lagares MA, Castanheira PN, Amaral DC, Vasconcelos AB, Veado JC, Arantes RM, Stahlberg R.The aim of the present study was to evaluate the in vitro viability of equine embryos vitrified in three different solutions. Day 6 and 6.5 embryos were measured and morphologically evaluated. Only grade 1 or 2 morulae and early blastocysts were vitrified. Eighteen embryos were distributed in Group 1: 40 percent ethylene glycol in PBS, Group: 2 and 3: 40 percent ethylene glycol, 18 percent Ficoll, 0.3M sucrose or 0.3M trehalose in PBS, respectively. The vitrified embryos were loaded individually into 0.25 ml straws, which were cooled and immersed in liquid nitrogen. After warming at 20 degree ...
Recovery of mare oocytes on a fixed biweekly schedule, and resulting blastocyst formation after intracytoplasmic sperm injection.
Theriogenology    March 4, 2010   Volume 73, Issue 8 1116-1126 doi: 10.1016/j.theriogenology.2010.01.013
Jacobson CC, Choi YH, Hayden SS, Hinrichs K.Oocytes may be collected from live mares from either the stimulated preovulatory follicle or from all visible immature follicles. We evaluated the yield of mature oocytes, and of blastocysts after intracytoplasmic sperm injection (ICSI), for both follicle types. In Experiment 1, mares were assigned to Progesterone (1.2g biorelease progesterone weekly) or Control treatments. Transvaginal aspiration of all follicles was performed every 14 d. Overall, 596 follicles were aspirated, with a 54% oocyte recovery rate. There was no difference between treatments in number of follicles punctured (9.0 to ...
Successful transfer of day 10 horse embryos: influence of donor-recipient asynchrony on embryo development.
Reproduction (Cambridge, England)    November 30, 2009   Volume 139, Issue 3 575-585 doi: 10.1530/REP-09-0306
Wilsher S, Clutton-Brock A, Allen WR.A total of 78 day 10 horse embryos were transferred non-surgically to recipient mares that had ovulated 9, 7, 6, 5, 4, 3, 2 or 1 day after (negative asynchrony), on the same day (synchronous), or 2 or 4 days before (positive asynchrony) the donor (n=6 or 8 mares per group). Pregnancy rates between 100% (6/6) and 63% (5/8) were seen in recipient mares that were between +2 and -6 days asynchronous. Embryo survival to the heartbeat stage declined in recipients that were -7 days asynchronous and no embryos survived in recipients that were -9 days asynchronous. Irrespective of uterine asynchrony, c...
Hormonal manipulation of endometrial gland development in the horse.
Equine veterinary journal    November 26, 2009   Volume 41, Issue 7 617-618 doi: 10.2746/042516409x467886
Spencer TE.No abstract available
The pattern of embryonic fixation and its relationship to pregnancy loss in thoroughbred mares.
Reproduction in domestic animals = Zuchthygiene    October 1, 2009   Volume 45, Issue 5 e61-e67 doi: 10.1111/j.1439-0531.2009.01523.x
Sharma S, Davies Morel MC, Dhaliwal GS, Dadarwal D.Ultrasonographic pregnancy records of 195 mares from six Thoroughbred stud farms, over a period of 7 years were retrospectively analysed to assess the effect of various factors on embryonic vesicle (EV) fixation pattern and pregnancy outcome. Of the total of 746 pregnancies analysed, significantly (p < 0.01) more EV fixations were evident in the right uterine horn than in the left (53.35% vs 46.65% respectively). There was no significant effect of either, the side of ovulation, or age of the mare, on the side of EV fixation. However, EV fixation, was significantly (p < 0.001) more likely...
The first ovulation of the breeding season in the mare: the effect of progesterone priming on pregnancy rate and breeding management (hCG response rate and number of services per cycle and mare).
Animal reproduction science    August 21, 2009   Volume 118, Issue 2-4 265-269 doi: 10.1016/j.anireprosci.2009.08.008
Cuervo-Arango J, Clark A.The mare is a seasonally polyestrus breeder. In early spring, the mare enters a "transition period" between the anovulatory season and the first ovulation of the year. This period is characterized by irregular estrus cycles and high incidence of regressing dominant follicles. There is a belief that pregnancy rates resulting from the first ovulation of the season is lower than in subsequent ovulations, however this has never been studied critically. Progestagens are often used as an aid to manage the transition period. The objective of this study was to compare pregnancy rates of mares from the...
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