218 research outputs found
Uterine glands agenesia in the mare
No full textAll mammalian uteri contain endometrial glands that synthesize and secrete a substance termed histotroph, which is essential for early pregnancy nutrition. Because of that function, endometrial glands play a crucial role as regulators of survival and development of conceptus. In all mammalian females as well as in woman, endometrial glands undergo extensive hyperplasia and hypertrophy during pregnancy as a response to increasing demands of the developing conceptus for uterine histotroph. The process of endometrial glands development is called adenogenesis. Inhibition of the development of these glands through gene mutation or epigenetic strategies results in infertility.
Two cases of total intrauterine glands developmental agenesis are presented, which were diagnosed on the base of histopathological examination of endometrial biopsy specimens. Despite the fact that endometrial glands agenesis in mare is a rare condition, it is worth while to emphasize that the endometrial biopsy is currently the best diagnostic tool to recognize the above developmental disturbance. Clinicians should be aware, that the mare with normal reproductive organs during macroscopic clinical investigation, can present with uterine glands agenesis
In vitro evaluation by DAPI staining of fresh, cooled and vitrified donkey embryos
No full textThe aim of this study was to evaluate the viability of fresh, cooled and vitrified donkey embryos by the 4’,6’-diamidino-2-phenylindole dihydrochloride (DAPI) staining method, was previously used for the evaluation of embryo damage (described as the percentage of dead per total cells) in the donkey species (Panzani et al. Theriogenology 2012;77:563–9). Eighteen 7 to 8 days old, quality I donkey embryos, recovered by uterine lavages with Ringer Lactate (RL), were divided in 3 groups: 1) Fresh (n=6, 7-days old; FR); 2) Cooled for 24 hours in a 5 ml test tube filled with the same RL recovered at the first uterine lavage (n=6, 8-days old; RL24); 3) Vitrified using the Equine Vitrification Kit (Bioniche Animal Health, USA) and the technique described for the horse (Eldridge-Panuska et al. Theriogenology 2005;63:1308–19) (n=6, 7-days old; VIT). After 24 hours of refrigeration in Equitainer® (RL24), after thawing (VIT) or directly after being recovered (FR), embryos were washed three times in Emcare Holding Solution (EHS), measured, moved for 5 min into EHS containing 1μg/mL DAPI and washed three more times in EHS. Dead embryonic cells (DAPI +) were directly counted under a fluorescence microscope, while the total cell number was estimated as described previously (n= 0.0106d2 + 2.0542 d – 375.28; n= cell number, d= embryo diameter in μm) (Moussa et al., Proc 20th Scientific Meeting of the IETS 2004;160). The proportion of DAPI stained cells was compared using One-Way ANOVA and LSD post-hoc test.
The proportion of dead cells in the embryos of the 3 groups was lower than that reported to be the maximum limit for viability of horse embryos (Moussa et al. Theriogenology 2005;64:1619–32) in fresh embryos it was similar to that previously reported in donkeys (070.3 vs 0.91.17)(Panzani et al., 2011). The proportion of dead cells differed among groups (P<0.05): vitrified embryos were more damaged than fresh or refrigerated embryos (P<0.05). The transfer of fresh and vitrified donkey embryos was performed in previous studies and at 14 days resulted in pregnancy rates of 50% and of 36.4%, respectively, and to the birth of the first two live donkey foals after transfer of cryopreserved embryos (Panzani et al. J Equine Vet Sci 2012;32:419–9). The results of this study need to be extended with recipients’ pregnancy rates after transfer of a significant number of vitrified and refrigerated donkey embryos
Postmating Endometritis and Pregnancy Rate Were Not Affected by the Addition to Frozen-Thawed Semen of Filtered Seminal Plasma When Mares Without Evidence of Endometritis Were Artificially Inseminated Once 40 Hours Post-Gonadotropin-Releasing Hormone Treatment
No full textThe main aim of this study was to evaluate the effect of the addition of seminal plasma to frozen-thawed semen on post mating endometritis and embryo recovery rate in mares when only one AI is done. Forty hours following induction of ovulation, 15 fertile Standardbred mares were submitted to a single artificial insemination (AI) per cycle with frozen semen obtained from one of two different stallions, on two cycles, according 2 different protocols: routine AI (200-280x106 frozen-thawed spermatozoa in 2 ml); and seminal plasma AI, (200-280x106 of frozen-thawed spermatozoa in 2 ml to which 7.8 ml of frozen-thawed seminal plasma was added). Six and 20 hours after AI, mares were evaluated by ultrasound for the presence of uterine fluid. Six hours after AI, mares were also subjected to uterine lavage for the evaluation of the presence and number of inflammatory cells. Eight days after ovulation, pregnancy was diagnosed by embryo recovery. There was a significant effect of treatment on subjective motility, which was lower when seminal plasma was added (20%; IQR: 10) compared to undiluted semen (45%; IQR: 10) (P<0.05). There was no significant effect of stallion or treatment on post mating endometritis or embryo recovery rate. In the mares and conditions of this study, the addition of seminal plasma to frozen-thawed semen had no effect on post AI uterine inflammation and pregnancy rate
Update on Donkey Embryo Transfer and Cryopreservation
No full textDonkey species reproductive biotechnology studies had a dramatic increase after the Convention of Biological Diversity that took place in Rio de Janeiro (Brazil) in 1992. The mechanization of agriculture of the 20th century took most of the developed countries donkey breeds close to extinction, and after Rio, the development of effective reproductive programs to save them was encouraged. Moreover, the recent interest in donkey milk, meat, and skin products transformed the donkey in a potential productive animal leading to the research and dissemination of desired productive tracts into its population. Among reproductive biotechnologies, the production and cryopreservation of embryos has a key role due to the possibility of producing and stocking valuable genetics potentially forever. Each species gametes and embryos from each species need specific media and protocols due to their particular characteristics. After more than 10 years of unsatisfactory results, embryo transfer and cryopreservation in the donkey moved from horse protocols to procedures more specifically tailored to the donkey species. Nowaday, pregnancy rates of 50% and 36% after the transfer of fresh and vitrified embryos, respectively, are possible in this species
Effects of centrifugation and removal of seminal plasma on motility and viability of cooled preserved amiata donkey spermatozoa
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Transrectal and transabdominal ultrasonographic study of Amiata donkey pregnancy from day 150 to term
No full textRetrospective study of factors affecting multiple ovulations, embryo recovery, quality, and diameter in a commercial equine embryo transfer program
No full textIn this study, 198 donor mares of different breeds, ages, and reproductive category were inseminated with fresh, cooled and frozen or frozen and cooled semen at the embryo transfer station or in private artificial insemination centers during 10 breeding seasons. The results of this activity were retrospectively analyzed by Pearson Chi-square test and logistic regression to evaluate factors affecting multiple ovulations, embryo recovery, embryo quality, and embryo diameter. Out of the 661 cycles, 937 ovulations were recorded (mean ovulations/cycle: 1.42 ± 0.58). Ovulation rate and incidence of multiple ovulations were significantly affected by age, breed, and reproductive category. Uterine flushings for embryo recovery were performed between 7 and 10 days after ovulation and resulted in the recovery of 338 embryos (51.1% embryos/cycle and 36.1% embryos/ovulation, respectively). At least one embryo was recovered in 298 flushings (45.1%). The factors affecting embryo recovery were age, breed, reproductive category, type of semen, number of ovulations, and location of artificial insemination. Flushing protocol and day of flushing had no effect on embryo recovery. Age, type of semen, number of ovulations, and day of flushing had a significant influence on embryo diameter (N = 215). None of the factors included in the model had an effect on embryo quality distribution
Donkey jack (Equus asinus) semen cryopreservation: Studies of seminal parameters, post breeding inflammatory response, and fertility in donkey jennies
No full textThe aims of this study were (1) to evaluate motility parameters of donkey jack (jack; Equus asinus) semen cryopreserved in INRA-96 (INRA; IMV Technologies, France, 2% egg-yolk enriched) using either glycerol (GLY) or ethylene glycol (EG) as a cryoprotector; (2) to compare in vitro the postthaw re-extension with homologous seminal plasma (SPL) or INRA; (3) to compare fertility in donkey jennies (jennies; Equus asinus) timed artificially inseminated with jack semen cryopreserved using GLY or EG, re-extended with INRA; (4) to compare fertility in jennies timed artificially inseminated with jack semen cryopreserved using GLY re-extended with SPL, INRA, or not re-extended (NN); and (5) to describe some preliminary results of the inflammatory uterine response postbreeding. Semen from two jacks was collected and frozen in an INRA-2% egg yolk extender added of either 2.2% GLY or 1.4% EG. Postthaw motility was evaluated by a computer-assisted motility analyzer. Uterine inflammatory response and fertility were evaluated after artificial insemination (AI) of 13 jennies with frozen-thawed semen, either further extended with INRA (Group GLY-INRA, 13 cycles, and EG-INRA, 8 cycles), or with SPL (Group GLY-SPL, 13 cycles), or not re-extended (GLY-NN, 5 cycles). In each cycle, jennies were bred twice with 500 * 10(6) sperm cells (250 * 10(6) from each jack), at fixed times after induction of ovulation, and uterus was flushed at 6 and 10 h after first and second breeding, respectively. Cells in the recovered fluid were counted and distinguished as polymorphonuclear neutrophils (PMN) or other cell types. Total and progressive motility did not differ between cryoprotectants, but were higher when semen samples were re-extended in INRA, compared with SPL (P < 0.05). Pregnancy was diagnosed by transrectal palpation and ultrasonography examinations at 14 and 16 days postovulation. In 7/13 (53.8%) jennies and 12/39 (30.4%) cycles postbreeding intrauterine fluid accumulation was observed, with no differences between treatments (P < 0.05). Polymorphonuclear neutrophil numbers and concentrations were higher in the first flushing compared with the second, and PMN concentration was higher in GLY-SPL than in GLY-INRA (P < 0.05). Pregnancy rates in GLY-SPL, GLY-INRA, EG-INRA, and GLY-NN were 8/13, 3/13, 2/8, and 1/5, respectively. There was no significant difference either between the two cryoprotectants re-extended in INRA, or between re-extension groups. There was however a trend for GLY-SPL to improve pregnancy rates compared with GLY-INRA (P = 0.055). These results indicate that it is possible to obtain similar postthaw sperm motility and pregnancy rates using GLY or EG as a cryoprotectant for donkey semen, and that in the conditions of this study the re-extension in SPL of thawed semen before AI showed a trend toward the improvement of fertility and increased PMN concentration in uterine flushings. Copyright 2012 Elsevier Inc. All rights reserved
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