Journal articles on the topic 'Frozen and thawed'
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Huang, Shaohua, Christina Miao, Sammy Sun, and Sameh Toma. "Monozygotic twins resulted from frozen thawed blastocyst generated from frozen thawed egg." Cryobiology 85 (December 2018): 176–77. http://dx.doi.org/10.1016/j.cryobiol.2018.10.216.
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Hollinshead, F. K., L. Gillan, J. K. O'Brien, G. Evans, and W. M. C. Maxwell. "In vitro and in vivo assessment of functional capacity of flow cytometrically sorted ram spermatozoa after freezing and thawing." Reproduction, Fertility and Development 15, no. 6 (2003): 351. http://dx.doi.org/10.1071/rd03060.
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The effect of sex sorting and freeze–thawing on the viability and fertility of ram spermatozoa was investigated in the present study. Non-sorted (control) frozen–thawed spermatozoa had a higher motility and forwards progressive motility (FPM) than sorted frozen–thawed spermatozoa (60.9 ± 2.9% v. 57.0 ± 3.3% and 4.0 ± 0.1 v. 3.5 ± 0.1 FPM, respectively; P < 0.001) after incubation (6 h at 37°C). Sorted and non-sorted (control) frozen–thawed spermatozoa had similar acrosome integrity (73.7 ± 1.8% v. 75.2 ± 2.1%, respectively) after thawing and incubation. A greater proportion of sorted spermatozoa displayed chlortetracycline staining patterns that were characteristic of capacitation (22.0 ± 2.8%; P < 0.05) than non-sorted (control) spermatozoa (15.4 ± 2.6% B pattern) before freezing. Overall, more sorted frozen–thawed spermatozoa showed patterns characteristic of being acrosome reacted (12.8 ± 0.7%; P < 0.01) and less were uncapacitated (35.5 ± 0.6%; P < 0.05) than non-sorted (control) frozen–thawed spermatozoa (7.7 ± 0.8% and 38.6 ± 0.6% for AR and F pattern, respectively). Similar numbers of non-sorted (control) and sorted frozen–thawed spermatozoa migrated through artificial cervical mucus after 1 h (76.4 ± 11.9 v. 73.9 ± 11.9 spermatozoa, respectively). The distance travelled by the vanguard spermatozoon was also similar (56.9 ± 7.8 v. 38.6 ± 5.8 mm for control and sorted spermatozoa, respectively). Sorted and control frozen–thawed spermatozoa displayed a similar pattern of binding to, and release from, an oviduct epithelial cell monolayer (OECM), but sorted frozen–thawed spermatozoa were released more rapidly (P < 0.05) than non-sorted (control) frozen–thawed spermatozoa. The pregnancy rate was higher for ewes inseminated with 100 × 106 (commercial control) frozen–thawed spermatozoa (59%) than for 5, 10, 20 and 40 × 106 total sorted frozen–thawed spermatozoa (41% overall; P < 0.001). Insemination of 16 × 106 resulted in a higher pregnancy rate (31%) than 106 (17%; P < 0.05), but was similar to ewes that received 4 × 106 sorted frozen–thawed spermatozoa (24%). Time of insemination (54, 58 and 62 h after sponge removal) had no effect on pregnancy rate. Pregnancy in gonadotrophin-releasing hormone-treated ewes was affected by insemination dose (P < 0.05) but not sperm type (sorted and non-sorted) or ram. Pregnancy was higher after insemination of 40 × 106 than 5 or 20 × 106 non-sorted (control) or sorted frozen–thawed spermatozoa (70%, 33% and 35%, respectively; P < 0.05). Sorted frozen–thawed spermatozoa may have a shorter viability within the female tract than non-sorted frozen–thawed spermatozoa.
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Diana, C., Elis Dihansih, and Dede Kardaya. "PHYSICAL AND CHEMICAL QUALITIES OF FROZEN BEEF WITHIN DIFFERENT THAWING METHOD." JURNAL PERTANIAN 9, no. 1 (May 18, 2018): 51. http://dx.doi.org/10.30997/jp.v9i1.1155.
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Different thawing metods were applied to frozen beef in order for evaluating both the physical and chemical qualities. The study used a completely randomized design with six treatments as follow:1) fresh beef as control, 2) frozen beef allowed at room temperature (27-300C) until internaltemperature of beef reached 00C (became unfrozen), 3) Frozen beef thawed at refrigerator temperature, i.e. 8-100C, 4) Frozen beef thawed at running water which its temperature range within 25-280C, 5) Frozen beef thawed by boiling water (1000C), and 6) Frozen beef thawed by hot water (<1000C). Every treatment was made in three replicates. Results of the study repealed that frozen beef thawed by running water, hot water, or boiling water resulted in better physical qualities than the one thawed by refrigerator temperature (P<0.05). All thawing methods did not significantly affect on chemical qualities of the beef (P>0.05). Moreover, all frozen beef showed similar chemical qualities to the fresh beef.
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Abie, Sisay Mebre, Ørjan Grøttem Martinsen, Bjørg Egelandsdal, Jie Hou, Frøydis Bjerke, Alex Mason, and Daniel Münch. "Feasibility of Using Electrical Impedance Spectroscopy for Assessing Biological Cell Damage during Freezing and Thawing." Sensors 21, no. 12 (June 16, 2021): 4129. http://dx.doi.org/10.3390/s21124129.
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This study was performed to test bioimpedance as a tool to detect the effect of different thawing methods on meat quality to aid in the eventual creation of an electric impedance-based food quality monitoring system. The electric impedance was measured for fresh pork, thawed pork, and during quick and slow thawing. A clear difference was observed between fresh and thawed samples for both impedance parameters. Impedance was different between the fresh and the frozen-thawed samples, but there were no impedance differences between frozen-thawed samples and the ones that were frozen-thawed and then stored at +3 °C for an additional 16 h after thawing. The phase angle was also different between fresh and the frozen-thawed samples. At high frequency, there were small, but clear phase angle differences between frozen-thawed samples and the samples that were frozen-thawed and subsequently stored for more than 16 h at +3 °C. Furthermore, the deep learning model LSTM-RNN (long short-term memory recurrent neural network) was found to be a promising way to classify the different methods of thawing.
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O'Brien, J. K., F. K. Hollinshead, G. Evans, and W. M. C. Maxwell. "332IN VIVO DEVELOPMENTAL CAPACITY OF IN VITRO-PRODUCED EMBRYOS DERIVED FROM SEX-SORTED AND RE-CRYOPRESERVED FROZEN-THAWED RAM SPERM." Reproduction, Fertility and Development 16, no. 2 (2004): 286. http://dx.doi.org/10.1071/rdv16n1ab332.
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The ability to sort and re-freeze frozen-thawed sperm would significantly increase the potential application of sperm sexing technology to species management. Frozen-thawed, sorted, re-frozen and then thawed ram sperm appear fully functional in vitro with blastocyst production greater than that for frozen-thawed, non-sorted sperm (Hollinshead FK et al. 2003 Theriogenology 59, 209 abst). The aim of this study was to evaluate the in vivo capacity of in vitro-produced embryos derived from frozen-thawed sperm after sorting and a second cryopreservation/thawing step. Frozen semen from 2 rams (n=3 ejaculates per ram) was used throughout. Post-thaw sperm treatments comprised (i) non-sorted (Control); (ii) sorted (Froz-Sort) and (iii) sorted, then re-frozen (Froz-Sort-Froz). X and Y sperm were separated using a high-speed sorter (SX MoFlo®, DakoCytomation, Fort Collins, CO, USA) after incubation with Hoechst 33342 and food dye to eliminate nonviable sperm. Reanalysis revealed high levels (mean±SEM) of purity for X- and Y-enriched samples for all treatments (89±1.2%). At Day 6 post-insemination, 2 embryos (blastocyst stage or greater) were transferred per recipient. Data were analyzed by chi-square and Fisher Exact Test. In vivo embryo survival was similar across sperm treatments (28/64, 43.8% overall) and 20 of 23 (87.0%) sexed lambs were of the predicted sex (Table 1). These results demonstrate high in vivo developmental capacity of in vitro-produced sexed embryos derived from frozen-thawed ram sperm after sorting and a second cryopreservation/thawing step, and increase the potential application of sperm sexing technology. Research support by XY, Inc., Australian Research Council and Zoological Parks Board of NSW. Table 1 In vivo survival of transferred in vitro produced embryos derived from frozen-thawed non-sorted (Control), frozen-thawed and sorted (Froz-Sort) and frozen-thawed, sorted, then frozen-thawed (Froz-Sort-Froz) ram sperm.
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de Graaf, S. P., G. Evans, W. M. C. Maxwell, and J. K. O'Brien. "In vitro characteristics of fresh and frozen - thawed ram spermatozoa after sex sorting and re-freezing." Reproduction, Fertility and Development 18, no. 8 (2006): 867. http://dx.doi.org/10.1071/rd06061.
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The in vitro function of sex-sorted, frozen–thawed ram spermatozoa derived from fresh or frozen semen was investigated. Sorted, frozen–thawed spermatozoa had higher (P < 0.05) motility, viability, acrosome integrity and mitochondrial activity than non-sorted, frozen–thawed controls immediately following thawing and after incubation at 37°C for 3 and 6 h. Similarly, frozen–thawed, sorted, re-frozen–thawed spermatozoa outperformed (P < 0.05) non-sorted controls upon thawing (mitochondrial activity) and following a 3-h incubation (motility, viability/acrosome integrity and mitochondrial activity), but there were no differences after incubation for 6 h (P > 0.05). Velocity characteristics (computer assisted sperm assessment 0–6 h post-thaw) of sorted spermatozoa derived from either fresh or frozen semen remained inferior (P < 0.05) to non-sorted spermatozoa, as did their ability to penetrate artificial cervical mucus after thawing. Direct comparison of cryopreserved spermatozoa derived from either fresh or frozen semen revealed that frozen–thawed, sorted, re-frozen–thawed spermatozoa had comparable (P > 0.05) motility, viability/acrosome integrity, mitochondrial activity, average path velocity and oviducal binding capacity immediately post-thaw, but reduced (P < 0.05) quality after 3 and 6 h of incubation. These findings indicate that, under the tested in vitro conditions, sex-sorted spermatozoa derived from fresh semen are superior in some respects to those derived from frozen semen. Further, that the use of either technique, while reducing velocity characteristics and cervical mucus penetration, results in comparable, if not enhanced motility, membrane and mitochondrial function in the post-thaw population of spermatozoa when compared with non-sorted, frozen–thawed controls.
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Ito, J., Y. Seita, K. Fujiwara, K. Furukawa, S. Sugio, and N. Kashiwazaki. "155 ESTABLISHMENT OF AN IN VITRO FERTILIZATION PROTOCOL USING CRYOPRESERVED EPIDIDYMAL AND EJACULATED RAT SPERMATOZOA." Reproduction, Fertility and Development 24, no. 1 (2012): 189. http://dx.doi.org/10.1071/rdv24n1ab155.
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In rats, successful IVF with cryopreserved rat sperm has never been reported. The objective of the present study was to establish and improve the IVF protocol using epididymal and ejaculated rat spermatozoa after cryopreservation. At first, we examined whether cryopreserved ejaculated spermatozoa would be useful for IVF in Wistar rats. Capacitation-associated tyrosine phosphorylation was accelerated in frozen-thawed ejaculated sperm in a time-dependent manner. These frozen-thawed spermatozoa were co-cultured with cumulus–oocyte complexes in modified R1ECM for 10 h. The putative zygotes were transferred to R1ECM and then cultured up to 144 h. Although the rates of insemination and 2 pronucleus (2PN) formation were low (26.5 and 23.0%, respectively), most of 2PN oocytes were developed to the 2-cell stage (91.0%). A total of 44 embryos at the 2-cell stage derived from frozen-thawed ejaculated sperm were transferred to 5 recipient females and 21 pups (47.7%) were delivered. Next, we used frozen-thawed epididymal spermatozoa for IVF in Wistar rats. After thawing, intracellular cyclic adenosine monophosphate (cAMP), free cholesterol levels and capacitation-associated protein tyrosine phosphorylation levels of the sperm were assayed. Intracellular cAMP and free cholesterol levels in frozen-thawed epididymal sperm were maintained at a low level, suppressing capacitation-associated tyrosine phosphorylation. Treatment with a phosphodiesterase inhibitor, 3-isobutyl-1-methyl-xanthin (IBMX) dramatically increased cAMP and capacitation-associated tyrosine phosphorylation levels in frozen-thawed epididymal sperm. When the IBMX-treated frozen-thawed sperm was used for IVF, the proportions of 2PN and the development to blastocysts were significantly higher (approximately 40 and 20%, respectively) than those of frozen-thawed epididymal sperm treated without IBMX (approximately 10 and 3%, respectively). These embryos were developed to term at a high success rate (49%) equivalent to the rate obtained with IVF using fresh sperm (58%). Moreover, we tried to apply our IVF system to inbred rat strains [Fischer 344 (F344) and Brown-Norway (BN)]. We examined whether the IVF protocol was available for F344 and BN rats. Fresh and frozen-thawed sperm collected from cauda epididymides in F344 and BN were used for detection of capacitation-associated tyrosine phosphorylation. In fresh F344 sperm, capacitation-associated tyrosine phosphorylation was induced in a time-dependent manner. Although tyrosine phosphorylation was inhibited in frozen-thawed F344 sperm, it was dramatically accelerated by IBMX treatment as well as frozen-thawed Wistar sperm. However, tyrosine phosphorylation in fresh and frozen-thawed BN sperm was suppressed and the phosphorylation in frozen-thawed sperm was not improved by IBMX. Taken together, we developed an IVF protocol using cryopreserved rat sperm and our data suggest that the IVF system can be applied not only to Wistar rats but also to the F344 strain. This research was also partially supported by a research project grant awarded by the Azabu University Research Services Division.
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Jahn, Egbert. "Aufgetaut und wieder eingefroren." osteuropa 70, no. 12 (2020): 81. http://dx.doi.org/10.35998/oe-2020-0097.
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Donnez, Jacques, Jean Squifflet, and Marie-Madeleine Dolmans. "Frozen-Thawed Ovarian Tissue Retransplants." Seminars in Reproductive Medicine 27, no. 06 (October 5, 2009): 472–78. http://dx.doi.org/10.1055/s-0029-1241057.
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Fancsovits, Péter, János Urbancsek, László Fónyad, Anna Sebestyén, Gézáné †Csorba, Ádám Lehner, Zita Kaszás, János Rigó jr., and Attila Bokor. "Kezdeti tapasztalataink a petefészekszövet-fagyasztás bevezetésével." Orvosi Hetilap 157, no. 49 (December 2016): 1947–54. http://dx.doi.org/10.1556/650.2016.30582.
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Introduction: The oncological treatment may damage ovarian function. To prevent this, it is possible to cryopreserve the ovarian tissue, and to keep the samples for long-term storage. The frozen-thawed tissue could be retransplanted after chemo- or radiotherapy. Aim: The aim of our study was to examine the effect of cryopreservation on the viability of ovarian tissue. Method: We analyzed the survival of frozen-thawed donated ovarian tissues. The quality of the follicles and hormone production in fresh and frozen-thawed samples were compared. Results: Histological analysis showed that the number of viable follicles was reduced by 23% in the frozen-thawed samples. However, viable follicles still presented in post thawing ovarian tissues. Maximal estradiol production in frozen-thawed tissues was 908 pg/ml and hormone production was similar to the control tissues. The maximal progesterone production was 1.95 ng/ml post thawing, but these values were lower than the progesterone production of fresh tissues. Conclusions: The method of ovarian cryopreservation used in our laboratory was able preserve the viability of follicles in frozen-thawed ovarian tissues. Orv. Hetil., 2016, 157(49), 1947–1954.
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Gillan, L., G. Evans, and W. M. C. Maxwell. "The interaction of fresh and frozen-thawed ram spermatozoa with oviducal epithelial cells in vitro." Reproduction, Fertility and Development 12, no. 6 (2000): 237. http://dx.doi.org/10.1071/rd00032.
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In order to investigate the interaction of fresh and frozen–thawed spermatozoa with oviduct epithelial cells, spermatozoa were co-incubated with ovine oviduct epithelial cell monolayers (OECM) derived from either complete oviducts, at any stage of the oestrous cycle (Experiments 1 and 2), or from different regions of the oviduct at different stages of the cycle (Experiment 3). Fresh and frozen—thawed spermatozoa displayed different patterns of binding to, and release from, the OECM. Frozen—thawed spermatozoa immediately bound to the complete oviduct OECM and were released after 2 h. A small proportion of fresh spermatozoa bound immediately, increasing to a maximum after 2 h, and were gradually released thereafter. When only the cells that were released from the OECM were observed by chlortetracycline staining in Experiment 2, it was found that the presence of an OECM increased the number of capacitated fresh spermatozoa while decreasing the number of capacitated frozen–thawed spermatozoa. Overall, the OECM advanced the membrane state of both types of spermatozoa from uncapacitated to acrosome-reacted. Fresh and frozen—thawed spermatozoa bound to OECM derived from the cells of the isthmus and the ampulla in similar proportions. However, more spermatozoa were capacitated when incubated with OECM derived from isthmic rather than ampullary cells. Higher proportions of fresh spermatozoa bound to, and were acrosome-reacted following incubation with OECM derived from post- rather than pre-ovulatory tracts. Such differences were not observed for frozen—thawed spermatozoa. The findings reported in this study show that fresh and frozen—thawed spermatozoa behave differently when in contact with oviduct cells in vitro. This may be a consequence of the more advanced membrane state of the frozen spermatozoa upon thawing.
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Pathak, P. K., A. J. Dhami, and D. V. Chaudhari. "Correlations of Motion Characteristics and KinematicAttributes of Fresh and Frozen-thawed Spermatozoaof Gir Bulls." INDIAN JOURNAL OF VETERINARY SCIENCES AND BIOTECHNOLOGY 15, no. 01 (July 25, 2019): 9–13. http://dx.doi.org/10.21887/ijvsbt.15.1.2.
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This investigation was carried out on semen of three healthy mature breeding bulls of Gir breed to evaluate the interrelationships among sperm quality attributes of fresh and frozen-thawed semen assessed by Biovis CASA. The ejaculates (n = 24) having >75% initial motility were diluted @80 million sperm/mL using TFYG extender, filled in French mini straws, and were frozen using a programmable bio freezer after 4 hours of equilibration. The straws were thawed in a water bath at 37°C for 30 sec. The freshly diluted and frozenthawed samples were assessed for routine subjective tests and various motion characteristics/kinematics by Biovis CASA. The Pearson’s correlations for sperm motility and velocity/kinematic parameters of total motile sperm as well as of progressively motile sperm were studied in freshly diluted and frozen-thawed semen. In fresh semen, total motile sperm assessed by CASA had significant (p less than 0.05, 01) correlations with rapid progressive motile sperm (r = 0.46), wobbling index (r = 0.52) and dancing frequency (r = -0.43) in fresh semen. In frozen-thawed semen, it was significantly correlated only with linearity (r = 0.46). The rapid progressive motile sperm in both fresh (r = 0.41 to 0.92) and frozen-thawed (r = 044 to 0.88) semen, however, had significant correlations with most of their velocity traits. Further, the average path velocity (VAP), curvilinear velocity (VCL), straight line velocity (VSL), linearity (LIN), straightness (STR), wobbling (WOB), beat-cross frequency (BCF), amplitude of lateral head displacement (ALH), and dancing mean (DNM) of sperm showed significant positive or negative interrelationships among each other in both fresh (r = 0.41 to 0.91) as well as post-thawed (r = 0.44 to 0.90) semen. Moreover, the correlations of motility and kinematics parameters of total motile sperm in both fresh and frozen-thawed semen were highly significant with velocity/kinematics traits of only progressively motile sperm, and the velocity traits among only motile sperm were highly significantly interrelated in both fresh (r = 0.46 to 0.98) and frozen-thawed (r = 0.43 to 0.93) semen of Girbulls, though the magnitudes of correlations were lower in frozen-thawed semen as compared to fresh semen. Thus, CASA analysis offresh semen for motility and velocity traits could predict the post-thawed sperm motility and velocity/kinematics of bovine semen.
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Maxwell, W. M. C., G. Evans, S. T. Mortimer, L. Gillan, E. S. Gellatly, and C. A. McPhie. "Normal fertility in ewes after cervical insemination with frozen - thawed spermatozoa supplemented with seminal plasma." Reproduction, Fertility and Development 11, no. 2 (1999): 123. http://dx.doi.org/10.1071/rd99046.
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The effect of seminal plasma (SP) on the motility, capacitation status, penetration through cervical mucus and fertility of frozen–thawed ram spermatozoa was examined. In the presence of SP, motility of frozen–thawed spermatozoa was better (P<0.001) and there were more uncapacitated and less acrosome-reacted cells in comparison with controls (P<0.001). Frozen–thawed spermatozoa were also better able to penetrate cervical mucus after addition of SP. Addition of SP increased the percentage of ewes pregnant after insemination of frozen–thawed (39/94, 41.5% v. 51/92, 55.4%; P<0.05) but not fresh spermatozoa (34/55, 61.8% v. 42/58, 72.4% for 0 v. 30% SP in the resuspension medium). Moreover, SP improved pregnancy rates after cervical (14/50; 28% v. 25/49; 51%; P<0.05) but not intrauterine insemination (25/44; 56.8 v. 26/43; 60.5%) with frozen–thawed spermatozoa. In a second experiment, pregnancy rates were 30/45 (66.7%), 9/37 (24.3%) and 24/40 (60.0%) for ewes inseminated with frozen–thawed spermatozoa in the uterus (control), cervix without SP and cervix after supplementation with SP, respectively (P<0.01 for unsupplemented v. supplemented spermatozoa). These experiments demonstrate that impaired function of cryopreserved spermatozoa can be overcome by addition of SP, resulting in normal fertility after cervical AI.
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Peris-Frau, Patricia, Irene Sánchez-Ajofrín, Alicia Martín Maestro, Carolina Maside, Daniela Alejandra Medina-Chávez, Olga García-Álvarez, María del Rocío Fernández-Santos, et al. "Impact of Cryopreservation on Motile Subpopulations and Tyrosine-Phosphorylated Regions of Ram Spermatozoa during Capacitating Conditions." Biology 10, no. 11 (November 20, 2021): 1213. http://dx.doi.org/10.3390/biology10111213.
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The heterogeneous nature of ejaculates highlights the relevance of studying the behavior of different sperm subpopulations. Changes in sperm motility and the increase in tyrosine phosphorylation are key events that usually occur during capacitation and can be modified by the cryopreservation process. However, the relationship between both events remains poorly defined throughout capacitation in the different sperm subpopulations. Fresh and frozen-thawed spermatozoa were incubated in capacitating (CAP) and non-capacitating (NC) media up to 240 min. Sperm kinematics, tyrosine phosphorylation and mitochondrial activity were measured by the CASA system and imaging flow cytometry. Four motile sperm subpopulations (SP) were identified in fresh and frozen-thawed ram semen after the cluster analysis. Incubation under CAP conditions over time led to greater changes in the percentage of spermatozoa included in each subpopulation compared to NC conditions, being different between fresh and frozen-thawed spermatozoa. The SP1, characterized by slow spermatozoa, progressively increased after 15 min in frozen-thawed samples incubated in both media but not in fresh ones. The SP4, characterized by fast and non-linear spermatozoa, showed a marked increase during CAP, but not under NC conditions, occurring more rapidly in frozen-thawed spermatozoa. This subpopulation (SP4) was also the only one positively and strongly correlated with mitochondrial activity and all phosphorylated sperm regions during capacitation, either in fresh or frozen-thawed samples. Our results indicated that in vitro capacitation induced significant changes in the distribution of motile sperm subpopulations, being affected by cryopreservation. Notwithstanding, the subpopulation which probably represents hyperactivated-like spermatozoa (SP4) also increased in frozen-thawed samples, occurring faster and simultaneously to the increment of mitochondrial activity and tyrosine phosphorylation of different sperm regions.
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Segino, Miwa, Mario Ikeda, Fumiki Hirahara, and Kahei Sato. "In vitro follicular development of cryopreserved mouse ovarian tissue." Reproduction 130, no. 2 (August 2005): 187–92. http://dx.doi.org/10.1530/rep.1.00515.
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In a previous report, we showed that follicles isolated from frozen/thawed mouse ovarian tissues reached the mature follicle stage on the 12th day of culture. However, the developmental ability was lower than that of fresh ovarian tissue. The purpose of this study was to define a culture system with some technical modification for preantral follicles isolated from frozen/thawed ovarian tissue and to confirm cell injury. Ovaries obtained from three-week-old female mice were cryopreserved by the rapid freezing method. Preantral follicles isolated from frozen/thawed ovarian tissues were cultured for 12–16 days. The follicles were then stimulated with human chorionic gonadotropin. In vitro fertilization was performed on the released cumulus–oocyte complexes (COCs). Preantral follicle viability was assessed by supravital staining using Hoechst 33258. Using this stain cell death was found in part of the granulosa cells of a follicle obtained from frozen/thawed ovarian tissue. On the 14th and 16th days of culture, the diameters of follicles isolated from frozen/thawed ovaries were larger than on the 12th day of culture. The released COCs were fertilized and developed to the blastocyst stage in 15.8% (12/76) of the oocytes taken from the fresh group, and in 0% (0/73), 2.9% (2/69) and 19.1% (22/115) of the oocytes taken from the frozen/thawed group that had been cultured for 12, 14 and 16 days respectively. The preantral follicles isolated from frozen/thawed mouse ovarian tissues developed slowly compared with the freshly prepared preantral follicles. During prolonged culture from 12 to 16 days, these follicles obtained the potential to fertilize and develop to the blastocyst stage.
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Ahmad, M., N. Ahmad, A. Riaz, and M. Anzar. "71 BOVINE SPERM DEATH KINETICS: CHANGES IN MOTILITY, ACROSOMES, AND PLASMA MEMBRANE." Reproduction, Fertility and Development 25, no. 1 (2013): 183. http://dx.doi.org/10.1071/rdv25n1ab71.
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Extent and timing of alterations in structures and functions of sperm after its placement in the female reproductive tract are important for successful fertilization. To our knowledge, the few reports are available on the kinetics of alterations in bovine sperm structures and functions during pathway to their death. Therefore, the present study was conducted to determine the changes in motility, acrosome and plasma membrane asymmetry in fresh and frozen–thawed semen during incubation at 37°C over the period of 24 h. Semen was collected from 3 breeding beef bulls, pooled, and considered as one replicate (total replicates = 5). Each pooled semen sample was diluted in Tris-citric acid egg yolk glycerol extender (pH 6.8), cooled to +4°C over 90 min, and then cryopreserved by a programmable cell freezer. Fresh (pooled semen) and frozen–thawed semen were incubated at 37°C for 24 h. Each semen sample was evaluated for sperm motility with computer-assisted semen analysis and acrosomal integrity and plasma membrane asymmetry using fluorescein isothiocyanate-peanut agglutinin/propidium iodide and Annexin V/propidium iodide assays, respectively, at 0, 2, 4, 6, 12, and 24 h of incubation at 37°C, with a flow cytometer. Statistical analysis was conducted using PROC MIXED model in statistical analysis system as 2 (semen types) × 6 (times) factorial model, using time as repeated measure. Progressive motility was higher (P < 0.05) in fresh than in frozen–thawed semen until 6 h. Progressive motility declined (P < 0.05) below the threshold level (i.e. 30%) much later (12 h) in fresh as compared with frozen–thawed semen (2 h). However, acrosomal integrity and plasma membrane asymmetry deteriorated (P < 0.05) below threshold at the same time interval (2 h) in both fresh and frozen–thawed semen. Viable sperm (AN–/PI–) remained higher (P < 0.05) during the first 6 h in fresh than in frozen–thawed semen and declined (P < 0.05) below the threshold at 12 h in fresh and at 6 h in frozen–thawed semen. In fresh semen, the necrotic sperm (AN–/PI+) population increased (P < 0.05) over time and reached maximum (97%) at 24 h. In frozen–thawed semen, a mixed population of late apoptotic (53%) and necrotic (34%) sperm was found at 24 h. In conclusion, the alterations in sperm motility, acrosomes, plasma membrane integrity, and asymmetry were slower in fresh than in frozen–thawed semen. Fresh sperm followed necrosis and frozen–thawed sperm underwent necrosis and apoptosis-like pathways, respectively. This study was supported by the Canadian Commonwealth Scholarship Program by the Canadian Bureau for International Education (CBIE), and Agriculture and Agri-Food Canada.
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Okazaki, T., T. Akiyoshi, M. Kan, H. Teshima, and M. Shimada. "82 CRYOPRESERVATION OF BOAR EPIDIDYMAL SPERMATOZOA; ADDITION OF SEMINAL PLASMA TO THAWING SOLUTION IMPROVES REPRODUCTIVE PERFORMANCE BY ARTIFICIAL INSEMINATION." Reproduction, Fertility and Development 23, no. 1 (2011): 146. http://dx.doi.org/10.1071/rdv23n1ab82.
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Epididymal spermatozoa are one of the available male germ cells for cryopreservation. It has been reported that frozen–thawed porcine epididymal spermatozoa have a high fertilization competence in vitro as compared with that in ejaculated one. However, there is little information about reproductive performance, such as conception rate or litter size, after artificial insemination (AI) using frozen–thawed epididymal spermatozoa. Recently, we demonstrated that the addition of seminal plasma to thawing solution improves membrane and acrosomal integrity, and enhanced both in vivo and in vitro fertilizing activity of frozen–thawed ejaculated spermatozoa. Moreover, the injection of seminal plasma to uterus with frozen–thawed spermatozoa significantly increased the number of implantation site (Okazaki et al. 2009 Theriogenology 71, 491–498). Thus, to apply those positive functions of seminal plasma to AI using frozen–thawed epididymal sperm, in this study, we added seminal plasma to thawing solution and then analysed the sperm functions including AI test using frozen–thawed epididymal spermatozoa. Epididymal spermatozoa collected by flushing caudal epididymis were frozen as described in our previous study (Okazaki et al. 2009). Frozen-spermatozoa were thawed in Modena solution with or without different percentages of seminal plasma. Protein tyrosine phosphorylation as a marker of capacitation was detected by western blotting. To examine the reproductive performance, the sows of natural oestrus were artificially inseminated two times (5 × 109 50 mL–1 per injection). When the frozen–thawed ejaculated or epididymal sperm was incubated up to 6 h, the motility of epididymal sperm was significantly higher than that of ejaculated sperm (19.6 v. 37.6%). However, the acrosomal membrane was damaged in epididymal sperm group at 3-h incubation period (15.2 v. 36.0%). The addition of seminal plasma [0, 10, 15, 20% (v/v)] in Modena solution protected the acrosomal injury (3 h; 35.2, 19.5, 15.6, 14.6%) and maintained high rate of motility (6 h; 38.8, 48.8, 62.5, 60.0%) in a dose-dependent manner. Furthermore, the addition of seminal plasma suppressed the expression of the 15 kDa phosphoprotein (early capacitation status), and the maximum effect was detected at 15% (v/v) seminal plasma. When the frozen–thawed epididymal spermatozoa with 15% (v/v) seminal plasma were artificially inseminated to swine (n = 15), the conception rate and the mean number of litter size were increased as compared with control (93 v. 43%, 10.0 v. 5.0). From these results, we concluded that the addition of seminal plasma to thawing solution was a beneficial method for artificial insemination using frozen–thawed epididymal spermatozoa in the pig. This work was supported by the Programme for Promotion of Basic and Applied Researches for Innovations in Bio-oriented Industry, and JST-Grant (No. 12-068 and No. 12-104).
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Borges-Silva, Juliana C., Márcio R. Silva, Daniel B. Marinho, Eriklis Nogueira, Deiler C. Sampaio, Luiz Orcírio F. Oliveira, Urbano G. P. Abreu, Gerson B. Mourão, and Roberto Sartori. "Cooled semen for fixed-time artificial insemination in beef cattle." Reproduction, Fertility and Development 28, no. 7 (2016): 1004. http://dx.doi.org/10.1071/rd14185.
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This study evaluated the use of cooled semen in a fixed-time artificial insemination (FTAI) program compared with frozen–thawed semen to improve pregnancy rates in beef cattle. Ejaculates of three bulls were collected and divided into two treatments: (1) frozen–thawed semen and (2) cooled semen. Egg-yolk extender without glycerol was used for the cooled semen treatment. Straws (25 × 106 spermatozoa) were submitted to cooling for preservation at 5°C for 24 h, after which FTAI was performed. Nelore cows (n = 838) submitted to FTAI were randomly inseminated using frozen–thawed semen or cooled semen. There was a 20% increase in the pregnancy per AI (P AI–1) using cooled semen compared with frozen–thawed semen (59.9 ± 4.7 vs 49.4 ± 5.0%; P < 0.005). There was no difference in P AI–1 among the bulls (P = 0.40). The frozen–thawed semen had fewer functional spermatozoa than did the cooled semen when evaluated by sperm motility (61.7 vs 81.0%), slow thermoresistance test (41.7 vs 66.7%) and hypoosmotic swelling test (38.3 vs 53.7%; P < 0.05). The percentage of sperm abnormalities did not differ between the freeze–thawing and cooling processes (18.6 vs 22.1%; P > 0.05). Because there was less damage to spermatozoa and improvement in P AI–1, the use of cooled semen instead of frozen–thawed semen is an interesting approach to increase reproductive efficiency in cattle submitted to a FTAI protocol.
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Jabbour, HN, and G. Evans. "Fertility of superovulated ewes following intrauterine or oviducal insemination with fresh or frozen-thawed semen." Reproduction, Fertility and Development 3, no. 1 (1991): 1. http://dx.doi.org/10.1071/rd9910001.
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Two experiments were conducted with mature Merino ewes to investigate the effects of time and site of insemination of fresh and frozen-thawed semen on the fertility of superovulated ewes. In Experiment 1, each ewe was treated with an intravaginal progestagen sponge and PMSG and/or FSH. They were inseminated in the uterus with fresh or frozen-thawed semen (approximately 100 x 10(6) motile sperm) at 24, 44 or 64 h after sponge withdrawal. Ova were recovered at 88 h after sponge withdrawal and classified as fertilized if they had pronuclei or had cleaved. Mean fertilization rates of recovered ova were 60.0, 93.7 and 87.8% for fresh semen and 46.1, 98.2 and 26.1% for frozen-thawed semen at each of the insemination times (24, 44 and 64 h) respectively. Overall, fertilization rates were higher for fresh semen than for frozen-thawed semen (P less than 0.01), but there was an interaction with time of insemination (P less than 0.01). Following insemination with frozen-thawed semen at 64 h, only 61% of the fertilized ova developed to the 2- to 4-cell stage by the time of embryo recovery; this was less than in any of the other groups (P less than 0.05). In Experiment 2, ewes were inseminated with 100 x 10(6) motile fresh or frozen-thawed semen in the uterus or in the oviducts at 64 h after sponge withdrawal.(ABSTRACT TRUNCATED AT 250 WORDS)
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Clulow, J. R., G. Evans, W. M. C. Maxwell, and L. H. A. Morris. "Evaluation of the function of fresh and frozen - thawed sex-sorted and non-sorted stallion spermatozoa using a heterologous oocyte binding assay." Reproduction, Fertility and Development 22, no. 4 (2010): 710. http://dx.doi.org/10.1071/rd09033.
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The aim of the present study was to evaluate the potential oocyte binding ability and functional integrity of fresh or frozen–thawed, sex-sorted or non-sorted stallion spermatozoa. In the absence of effective IVF procedures in the horse, a heterologous sperm-binding assay was used as an indicator of fertilising capacity to assess differences in the ability of stallion spermatozoa to bind to bovine oocytes. The functional integrity of four treatment groups was assessed: (1) fresh non-sorted spermatozoa; (2) fresh sex-sorted spermatozoa; (3) frozen–thawed non-sorted spermatozoa; and (4) frozen–thawed sex-sorted spermatozoa. Spermatozoa found in association with the zona pellucida of the bovine oocytes were deemed ‘attached’ or ‘bound’ depending on their characterisation as either acrosome intact or acrosome reacted, respectively. Significantly less frozen–thawed spermatozoa were found attached to the oocytes compared with fresh spermatozoa. No significant differences were identified between the number of attached sex-sorted and non-sorted frozen–thawed spermatozoa. However, significantly more sex-sorted than non-sorted fresh spermatozoa were found attached to the oocytes after 1 h coincubation, although after 3 h coincubation this difference was no longer apparent. In conclusion, sex-sorted fresh and frozen–thawed stallion spermatozoa are functionally capable of attaching and binding to bovine oocytes in vitro. Furthermore, fresh sex-sorted spermatozoa attach better than non-sorted spermatozoa, suggesting that they have a more advanced capacitation-like status.
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Suzuki, Sayuri, and Kahei Sato. "The fertilising ability of spermatogenic cells derived from cultured mouse immature testicular tissue." Zygote 11, no. 4 (November 2003): 307–16. http://dx.doi.org/10.1017/s0967199403002363.
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There are many reports about the in vitro culture of spermatogenic cells, but no-one has succeeded in inducing the differentiation from spermatogonia to intact sperm. Also the study of in vitro testicular tissue culture has hardly advanced. We studied the culture of mouse immature testicular tissue derived from 5-day-old mice. We aimed to achieve the differentiation of spermatogenic cells in order to observe spermatogenesis in testicular tissue in vitro. We also froze mature testicular tissue and immature testicular tissue cultured for 2 weeks. Furthermore, spermatogenic cells differentiated by culturing were injected into metaphase II oocytes to determine whether these differentiated cells and frozen-thawed testicular tissue have fertilising and developmental ability. Under the culture conditions employed, secondary spermatocytes and a few round spermatids differentiated from spermatogonia were observed in the immature testicular tissue cultured for 2 weeks. When spermatogenic cells derived from cultured immature testicular tissue, cultured frozen immature testicular tissue and frozen-thawed mature testicular tissue were injected into ooplasm, the oocytes were fertilised and fertilised oocytes developed to the 8-cell stage. We suggest that spermatogenic cells derived from cultured immature testicular tissue have fertilising and developmental abilities equivalent to that of sperm. Also these abilities of spermatogenic cells obtained from cultured frozen immature testicular tissue and frozen-thawed mature testicular tissue were better than those of the same cells before freezing.
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Šimoniová, A., B. A. Rohlík, T. Škorpilová, M. Petrová, and P. Pipek. "Differentiation between fresh and thawed chicken meats." Czech Journal of Food Sciences 31, No. 2 (April 18, 2013): 108–15. http://dx.doi.org/10.17221/127/2012-cjfs.
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Adulteration of fresh meat and its possible substitution with frozen and thawed meat represents a problem, since thawed meat has not only lower sensory qualities than fresh meat but also a lower price. Commercial enzymatic kits seem to be a solution to reveal such unfair practises and were applied to detect the activity of specific enzymes, citrate synthase, mitochondrial enzymes that are released from the organelles destroyed by frost. We determined, whether the meat of slaughtered chicken was fresh or frozen/thawed, and to provide convincing results. The absolute results vary with the type of meat and depend on the enzyme used. However, the enzyme activity in the exudates of frozen/thawed meat is always higher than in fresh meat. This value further increases with each subsequent freezing cycle. The determination of citrate synthase activity was done only in the exudate released from the examined meat samples. However, to determine the enzymes activity directly in unpacked meat, which have not released any exudate, is the subject of further research.
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Magarey, G., J. Herrick, K. Thiangtum, W. Tunwattana, and W. Swanson. "284 COMPARATIVE FERTILITY OF FRESHLY-COLLECTED VERSUS FROZEN - THAWED SPERMATOZOA FOR IN VITRO FERTILIZATION IN THE FISHING CAT (PRIONAILURUS VIVERRINUS)." Reproduction, Fertility and Development 18, no. 2 (2006): 249. http://dx.doi.org/10.1071/rdv18n2ab284.
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Wild populations of fishing cats (Prionailurus viverrinus) in Southeast Asia are in decline, primarily due to habitat loss. Because the fishing cat population in North American zoos is small (n = 69) and inbred (F = 0.17) with relatively low genetic variation (86%), infusion of new founder genes from Asia is a conservation priority. Importation of cryopreserved semen for use with IVF and ET may offer one alternative to the international transport of living animals. In this study, our objectives were to (1) compare motility longevity of fresh vs. frozen-thawed fishing cat spermatozoa in two culture media, (2) evaluate ovarian responses to exogenous gonadotropins, and (3) assess development of IVF embryos produced with fresh vs. frozen-thawed spermatozoa. Raw semen was collected via electroejaculation from male fishing cats (n = 4), divided into groups, and washed. Two sperm pellets were resuspended in either Ham's F10 medium (HF10; with 5% FBS) or our feline optimized culture medium (FOCM; with 0.4% BSA); another pellet was diluted in TEST egg yolk, cooled to 5�C over 3 h, glycerated (4%), and cryopreserved in straws over LN2 vapor. Frozen sperm samples were thawed, washed, and diluted in either HF10 or FOCM. Fresh and frozen-thawed sperm motility (percent motile, rate of forward progress) in each medium (10 � 106 motile sperm/mL) was assessed (at 0, 1, 3, and 6 h) in microdrops under oil during culture (38�C; 6% CO2 in air). Female fishing cats (n = 10) were treated with exogenous gonadotropins (150 IU eCG, 100 IU hCG, 85-h interval) and ovarian follicles were aspirated laparoscopically. Recovered oocytes were inseminated with fresh (2 � 105 motile sperm/mL) or frozen-thawed (5 � 105 motile sperm/mL) spermatozoa in FOCM microdrops; resulting embryos were either cryopreserved or cultured in FOCM (with 5% FBS added at 72 h post-insemination) for 7 days. Sperm motility over time did not differ (P > 0.05) between media for either fresh or frozen-thawed samples; however, across media, frozen-thawed sperm motility was lower (P < 0.05) and declined faster (P < 0.05) compared to fresh spermatozoa. Females produced an average (�SEM) of 9.8 � 2.9 mature ovarian follicles, allowing recovery of 7.3 � 2.6 high-quality oocytes per female. Oocyte cleavage percentage at 42 h p.i. was lower (P < 0.05) with frozen-thawed spermatozoa (38%, 11/29) compared to freshly collected spermatozoa (68%, 17/25). Overall, 35% (6/17) of cultured embryos developed to blastocysts with no difference (P > 0.05) between embryos produced with frozen-thawed (4/11) vs. fresh (2/6) spermatozoa. Although fishing cat sperm motility and fertility appear compromised after cryopreservation, our results demonstrate the ability of frozen-thawed spermatozoa to produce IVF embryos that are capable of developing to blastocyst stage in vitro. This work was supported by (NIH RR015388).
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Cattral, Mark S., Jonathan R. T. Lakey, Garth L. Warnock, Norman M. Kneteman, and Ray V. Rajotte. "Effect of Cryopreservation on the Survival and Function of Murine Islet Isografts and Allografts." Cell Transplantation 7, no. 4 (July 1998): 373–79. http://dx.doi.org/10.1177/096368979800700405.
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We compared the efficacy of fresh and frozen/thawed islets by determining the minimum number required to consistently reverse diabetes in mice. Defined numbers of islets, isolated from Balb/c (H-2d) and CBA/J (H-2k) mice, were transplanted into streptozotocin-induced diabetic Balb/c mice. Frozen/thawed grafts were cooled slowly to −40°C, stored at −196°C, and thawed rapidly. At 100 days after transplantation, isografts were recovered for measurement of insulin content. Mean (±SD) recovery of cryopreserved islets after thawing was 80 ± 3% (range 67–89%). For both fresh and frozen/thawed isografts and allografts, 200 islets were required to establish normoglycemia. The degree of metabolic function provided by equivalent quantities of fresh and frozen/thawed grafts was similar; and all normoglycemic isograft recipients remained so until graft nephrectomy. The insulin content of fresh and frozen/thawed isografts containing 200 and 300 islets were 151 ± 25 and 126 ± 8 mU and 259 ±36 and 278 ± 20 mU, respectively. Among allograft recipients, median survival ranged from 15 to 20 days, and was not influenced by cryopreservation or graft size. The results of this study demonstrate a high rate of recovery of viable islets following cryopreservation. The function of equivalent quantities of fresh and cryopreserved islet isografts and allografts in nonimmunosuppressed recipients is similar.
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Wang, Tian, Peng Li, Jingming Hou, Zhanbin Li, Zongping Ren, Shengdong Cheng, Guoce Xu, Yuanyi Su, and Feichao Wang. "Response of the Meltwater Erosion to Runoff Energy Consumption on Loessal Slopes." Water 10, no. 11 (October 26, 2018): 1522. http://dx.doi.org/10.3390/w10111522.
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Soil properties are influenced by freeze-thaw, which in turn influences soil erosion. Despite this, only a few studies have investigated the impacts on soil hydrodynamic processes. The objective of this study was to evaluate the impact of soil freezing conditions on runoff, its energy consumption, and soil erosion. A total of 27 laboratory-concentrated meltwater flow experiments were performed to investigate the soil erosion rate, the runoff energy consumption, and the relationship between the soil erosion rate and runoff energy consumption by concentrated flow under combinations of three flow rates (1, 2, and 4 L/min) and three soil conditions (unfrozen, shallow-thawed, and frozen). The individual and combined effects of soil condition, flow rate, and runoff energy consumption on the soil erosion rate were analyzed. For the same flow rate, the shallow-thawed and frozen slope produced mean values of 3.08 and 4.53 times the average soil erosion rates compared to the unfrozen slope, respectively. The number of rills in the unfrozen soil slope were 4, 3, and 2 under the flow rate of 1, 2, and 4 L/min, respectively. The number of rills in the thawed-shallow and frozen soil slope were all 1 under the flow rate of 1, 2, and 4 L/min. The rill displayed disconnected distribution patterns on the unfrozen slope, but a connected rill occurred on the shallow-thawed and frozen slopes. The average rill width on unfrozen, thawed-shallow, and frozen soil slopes increased by 1.87 cm, 4.38 cm, and 1.68 cm as the flow rate increased from 1 L/min to 4 L/min. There was no significant difference in the rill length on the frozen slope under different flow rates (p > 0.05). The runoff energy consumption ranged from unfrozen > shallow-thawed > frozen slopes at the same flow rate. The soil erosion rate had a linear relationship with runoff energy consumption. The spatial distribution of the runoff energy implied that soil erosion was mainly sourced from the unfrozen down slope, shallow-thawed upper slope, and frozen full slope.
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Lehloenya, K. C., N. Mahoete, J. P. C. Greyling, and T. L. Nedambale. "131 EFFECT OF BREED AND FROZEN - THAWED RAM SEMEN ON IN VITRO FERTILIZATION AND OVINE EMBRYONIC DEVELOPMENT." Reproduction, Fertility and Development 23, no. 1 (2011): 170. http://dx.doi.org/10.1071/rdv23n1ab131.
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Ovine embryonic development was evaluated 8 days following in vitro fertilization, after using fresh or frozen–thawed Merino and indigenous (Pedi and Zulu) sheep semen. Semen used was collected twice weekly over a 3-month period with the aid of an electro-ejaculator. Following collection, semen samples were evaluated and semen with acceptable sperm motility and a percentage live sperm of 60% diluted with an egg yolk-based extender (Egg-Yolk Citrate). Semen samples were cryopreserved in straws with a programmable freezer to –130°C and then plunged into liquid nitrogen (–196°C) until used for IVF. Fresh and frozen–thawed semen was used to fertilize the matured oocytes in vitro. A total of 791 oocytes were fertilized using fresh semen and 802 oocytes fertilized using frozen–thawed semen. No significant differences were recorded between the fresh and frozen–thawed semen regarding the embryonic developmental stages. The performance of fresh and frozen–thawed semen followed the same trend, with the cleavage rate gradually declining with the progression in time and the embryonic developmental stage. The lowest developmental rate recorded was the occurrence of blastocyst formation, ranging between 0.4 ± 0.4% and 2.6 ± 1.0%. Regarding breed, no significant difference was observed from cleavage to the 2- to 4-cell stages. The use of fresh and frozen–thawed Zulu semen resulted in a significantly (P < 0.05) higher percentage of 8-cell development compared with the Pedi semen. However, the 8-cell embryonic stage recorded with the use of the Zulu ram semen (fresh and frozen–thawed), did not differ significantly from that of the Merino breed. No significant difference between the breeds regarding blastocyst formation was recorded. The overall cleavage rate, 2- to 4-cell, and blastocyst embryonic developmental stages following the use of fresh and frozen–thawed semen from the different rams were generally lower than those recorded by other researchers. The low blastocyst rates obtained warrant more research regarding the in vitro embryo production technique in order to improve the ovine blastocyst formation rate. The study was funded by the University of the Free State and conducted at the Germplasm Conservation and Reproduction Biotechnologies ARC.
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Han, Ae Ra, Chan Woo Park, Hyoung-Song Lee, Kwang Moon Yang, In Ok Song, and Mi Kyoung Koong. "Blastocyst transfer in frozen-thawed cycles." Clinical and Experimental Reproductive Medicine 39, no. 3 (2012): 114. http://dx.doi.org/10.5653/cerm.2012.39.3.114.
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TOYAMA, Yoshiro, and Yoneto ITOH. "Ultrastructures of Frozen-thawed Boar Spermatozoa." Journal of Reproduction and Development 41, no. 5 (1995): j9—j13. http://dx.doi.org/10.1262/jrd.95-415j9.
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Graham, J. K., and E. Mocé. "Fertility evaluation of frozen/thawed semen." Theriogenology 64, no. 3 (August 2005): 492–504. http://dx.doi.org/10.1016/j.theriogenology.2005.05.006.
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TELIS, V. R. N., and T. G. KIECKBUSCH. "Viscoelasticity of Frozen/Thawed Egg Yolk." Journal of Food Science 62, no. 3 (May 1997): 548–50. http://dx.doi.org/10.1111/j.1365-2621.1997.tb04427.x.
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Sofikitis, N., Y. Yamamoto, and I. Miyagawa. "PREGNANCIES WITH FROZEN/THAWED TESTICULAR SPERMATOZOA." Japanese Journal of Urology 87, no. 2 (1996): 208. http://dx.doi.org/10.5980/jpnjurol.87.208_2.
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Roman, L. T., and P. I. Kotov. "Viscosity of Frozen and Thawed Soil." Soil Mechanics and Foundation Engineering 53, no. 1 (March 2016): 19–23. http://dx.doi.org/10.1007/s11204-016-9358-8.
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Reis, MM. "Near infrared spectroscopy (Vis-NIRS) applied to differentiation between chilled and frozen/thawed meat." NIR news 28, no. 7 (October 9, 2017): 10–15. http://dx.doi.org/10.1177/0960336017736246.
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The ability of Vis-NIRS to differentiate between only chilled and frozen and thawed meat from pork, lamb, beef and goat is investigated in this study. Samples were purchased as retail ready package from seven different local supermarkets. Partial least squares discriminant analysis and double cross-validation were used for analysis of the data. The discrimination between the two groups achieved an accuracy of 93%. There is 92% probability that a chilled sample is predicted correctly as chilled and 96% that frozen/thawed is a true frozen/thawed event, which demonstrates reasonable performance, i.e. independent of the species (pork, lamb, beef and goat) and source. It was possible to detect the difference between only chilled and frozen/thawed meat. The regression coefficients suggest that the differences between the two treatments are likely to be associated to changes in structure and chemical composition of samples due to the process of freezing/thawing.
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Underwood, S. L., R. Bathgate, D. C. Pereira, A. Castro, P. C. Thomson, W. M. C. Maxwell, and G. Evans. "Embryo production after in vitro fertilization with frozen-thawed, sex-sorted, re-frozen-thawed bull sperm." Theriogenology 73, no. 1 (January 2010): 97–102. http://dx.doi.org/10.1016/j.theriogenology.2009.08.005.
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Kim, M. K., S. H. Lee, H. S. Lee, I. O. Song, J. T. Seo, and Y. S. Park. "Pregnancy and delivery rates of frozen-thawed embryos fertilized with fresh- or frozen-thawed testicular sperm." Fertility and Sterility 94, no. 4 (September 2010): S112. http://dx.doi.org/10.1016/j.fertnstert.2010.07.461.
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Galkin, Aleksandr. "Calculation of the Fourier criterion in predicting the thermal regime of thawed and frozen dispersed rocks." Арктика и Антарктика, no. 3 (March 2022): 1–10. http://dx.doi.org/10.7256/2453-8922.2022.3.38555.
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The purpose of this work was to determine the range of changes in the Fourier criterion (number) when predicting the thermal regime of dispersed rocks in thawed and frozen state. And, also an assessment of the possibility of averaging the thermophysical characteristics of rocks to obtain universal values of Fourier numbers. To achieve this goal, an assessment of the influence of the thermophysical properties of dispersed rocks on the range of changes in the values of the Fourier number used in thermal calculations of technical objects of the cryolithozone is made. The calculation formulas took into account the functional dependence of the coefficient of thermal conductivity, density and specific heat capacity of rocks on humidity (iciness) in the thawed and frozen state. As an example, a mixture of quartz sandstone with water in a thawed and frozen state is considered when the ice content changes from zero (dry quartz sandstone) to full moisture saturation. It is established that the range and nature of the change in Fourier numbers for thawed and frozen dispersed rocks, depending on their humidity (iciness), differs significantly, not only quantitatively, but also qualitatively: for thawed dispersed rocks, the Fourier number decreases with increasing humidity, and for frozen rocks increases. The possibility of averaging the thermophysical characteristics of rocks to obtain universal values of Fourier numbers has been evaluated. It is shown that the use of universal Fourier numbers leads to a significant error for both thawed and frozen rocks and their use in thermal calculations with annual temperature fluctuations is impractical.
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Shaw, Lisa, Sharon F. Sneddon, Daniel R. Brison, and Susan J. Kimber. "Comparison of gene expression in fresh and frozen–thawed human preimplantation embryos." REPRODUCTION 144, no. 5 (November 2012): 569–82. http://dx.doi.org/10.1530/rep-12-0047.
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Identification and characterisation of differentially regulated genes in preimplantation human embryonic development are required to improve embryo quality and pregnancy rates in IVF. In this study, we examined expression of a number of genes known to be critical for early development and compared expression profiles in individual preimplantation human embryos to establish any differences in gene expression in fresh compared to frozen–thawed embryos used routinely in IVF. We analysed expression of 19 genes by cDNA amplification followed by quantitative real-time PCR in a panel of 44 fresh and frozen–thawed human preimplantation embryos. Fresh embryos were obtained from surplus early cleavage stage embryos and frozen–thawed embryos from cryopreserved 2PN embryos. Our aim was to determine differences in gene expression between fresh and frozen–thawed human embryos, but we also identified differences in developmental expression patterns for particular genes. We show that overall gene expression among embryos of the same stage is highly variable and our results indicate that expression levels between groups did differ and differences in expression of individual genes was detected. Our results show that gene expression from frozen–thawed embryos is more consistent when compared with fresh, suggesting that cryopreserved embryos may represent a reliable source for studying the molecular events underpinning early human embryo development.
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Colleoni, S., M. Spinaci, R. Duchi, B. Merlo, C. Tamanini, G. Lazzari, G. Mari, and C. Galli. "262 ICSI OF EQUINE OOCYTES WITH SEX-SORTED FROZEN-THAWED SEMEN RESULTS IN LOW CLEAVAGE RATE BUT NORMAL EMBRYO DEVELOPMENT AND PREGNANCIES." Reproduction, Fertility and Development 21, no. 1 (2009): 228. http://dx.doi.org/10.1071/rdv21n1ab262.
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Sorting of sperm by flow cytometer has allowed selection of offspring of predetermined sex in several species by artificial insemination, although the success rate is often lower than with non-sexed semen. In horses, the problem was partially overcome with hysteroscopic insemination using sex-sorted fresh sperm. However, when sex-sorted frozen–thawed sperm were used the pregnancy rate was heavily reduced in comparison with non-sexed frozen–thawed semen. Because it has been demonstrated that in vitro assisted reproductive techniques, namely intracytoplasmic sperm injection (ICSI), has permitted live foals to be obtained using sperm with low fertility in the field, in this study we investigated the possibility of using ICSI with sexed-sorted frozen–thawed sperm for equine embryo production in vitro. Briefly, semen was collected from two Standardbred stallions of proven fertility (Stallions A, B), sorted using a MoFlo SX flow cytometer and frozen (Johnson LA and Welch GR 1999 Theriogenology 52, 1323–1341). Sex-sorted and control non-sexed frozen semen (two stallions of in vitro proven fertility: C, D) was thawed, centrifuged on a Percoll gradient, washed and diluted 1:1 in PVP before ICSI. Oocytes were collected from ovaries of slaughtered mares and matured in vitro. Metaphase II oocytes were injected with sperm, subsequently cultured up to the blastocyst stage and frozen conventionally in 10% of glycerol (Galli et al. 2002 Theriogenology 58, 713–715). Six embryos from sexed-sorted sperm were thawed and non-surgically transferred in naturally cycling synchronous recipient mares. Results are summarized in Table 1. Overall, 70 and 58 (stallion A, B) and 30 and 15 (stallion C, D) oocytes were injected with sex-sorted or control frozen–thawed sperm, respectively. Mean cleavage rates were 20.3% for sorted sexed sperm and 71.1% for control, showing a significantly lower cleavage rate for sexed sperm. This difference was reflected in the number of blastocysts obtained (4.7% v. 20.0%). From the 6 frozen–thawed embryos derived from sexed sperm, that were transferred, 4 pregnancies resulted. One pregnancy was lost around 21 days, a second was pharmacologically aborted, and two were maintained (one from male and one from female sorted semen are currently in the 11th month of gestation). In conclusion, this study demonstrates that ICSI with sex-sorted sperm can be used for producing equine blastocysts able to establish pregnancies at a high rate following embryo transfer. However, the overall efficiency of the system is limited due to the very low cleavage rate obtained with sexed-sorted frozen–thawed sperm. Table 1.Development of embryos produced by ICSI with sorted and non-sorted frozen–thawed semen This work was supported by an RFO (ex 60%) and Camera di Commercio Cremona grant. The Authors wish to thank Società Italiana Produttori Sementi.
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Jankovičová, Jana, Michal Simon, Jana Antalíková, and Ľubica Horovská. "Acrosomal and viability status of bovine spermatozoa evaluated by two staining methods." Acta Veterinaria Hungarica 56, no. 1 (March 1, 2008): 133–38. http://dx.doi.org/10.1556/avet.56.2008.1.14.
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Artificial insemination with frozen-thawed spermatozoa is commonly used in cattle breeding. A simple and fast procedure is needed for routine evaluation of the acrosomal status of frozen-thawed bovine sperm. Therefore, the purpose of this study was to test two staining procedures used to determine the viability and integrity of acrosome of frozen-thawed bovine spermatozoa. Double staining and Hoechst/FITC- Pisum sativum agglutinin (FITC-PSA) labelling were tested for evaluating the viability and acrosome reaction induced by calcium ionophore of bull spermatozoa. In our experiments no significant differences were detected in the frequency of acrosome-reacted sperm either by double staining (37.98%) or by FITC-PSA labelling (39.33%). The viability of sperm stained by the double staining method was 67.17%, and a higher portion of viable sperm (82.67%) was observed by staining with the Hoechst procedure (P < 0.01). On the basis of the results obtained it is concluded that both methods can be used for detecting the acrosome reaction of frozen-thawed bovine spermatozoa.
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GULDAGER, HELLE SKOV, NIELS BØKNÆS, CARSTEN ØSTERBERG, JETTE NIELSEN, and PAW DALGAARD. "Thawed Cod Fillets Spoil Less Rapidly Than Unfrozen Fillets When Stored under Modified Atmosphere at 2°C." Journal of Food Protection 61, no. 9 (September 1, 1998): 1129–36. http://dx.doi.org/10.4315/0362-028x-61.9.1129.
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The effect of two months of frozen storage at −20°C on the spoilage characteristics and shelf life of thawed and modified atmosphere packed (MAP) cod fillets stored at 2°C was studied. Thawed MAP cod fillets were compared with fresh cod fillets stored in CO2-containing modified atmospheres with and without added oxygen. The shelf life of 11 to 12 days in the fresh MAP cod was extended to more than 20 days in the thawed MAP cod at 2°C. This shelf life extension was most likely due to the inactivation of the spoilage bacterium Photobacterium phosphoreum during frozen storage as reflected both in Chemical analyses and sensory evaluation. In contrast to fresh MAP cod fillets no significant production of trimethylamine occurred and almost no amine odor and taste were detected during 20 days of chill storage of thawed MAP cod fillets. The use of frozen fillets as raw material not only provides a more stable product in MAP but also allows much greater flexibility for production and distribution. However, a slightly increased concentration of dimethylamine, a larger drip loss, and detection of weak frozen storage flavor were observed in the thawed MAP cod fillets.
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Kulíková, Barbora, Marta Oravcová, Andrej Baláži, Peter Supuka, and Peter Chrenek. "Factors affecting storage of Slovak native rabbit semen in the gene bank." Zygote 25, no. 5 (August 24, 2017): 592–600. http://dx.doi.org/10.1017/s0967199417000454.
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SummaryIn this study, fresh and frozen–thawed semen of Nitra and Zobor rabbit breeds were evaluated for potential inter-breed or inter-male differences in sperm quality traits. Individual male semen from four rabbits of each breed were diluted (v:v; 1:1) in a freezing medium composed of a commercial diluent, 16% of dimethyl sulphoxide (DMSO), 4% of Ficoll 70 and 2% of sucrose and frozen in liquid nitrogen vapours before being plunged into liquid nitrogen. Different motility traits, viability and plasma membrane integrity of fresh and frozen–thawed semen were evaluated in vitro using computer-assisted sperm analysis and flow cytometry. To evaluate the sperm fertilization ability, artificial insemination of fresh and frozen–thawed sperm was performed. Our results showed the effect of breed (P ≤ 0.05) on frozen–thawed sperm viability and plasma membrane integrity. Moreover, individual variability in semen quality among the rabbits was revealed (0.31 to 0.71 among quality traits). Our results thereby confirmed that the cryopreservation procedure could not ensure comparable sperm post-thaw survival for different breeds or males. Nevertheless, correlations between numbers of fresh total motile and progressively moving sperm and several quality parameters measured post thawing were revealed. Therefore, we suggest that the objective assessment of fresh rabbit sperm motility may be an effective indicator of frozen–thawed semen quality. Consequently, regular semen assessment is required in order to preserve good-quality insemination doses from native breeds.
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Urbano, M., J. Dorado, I. Ortiz, M. J. Galvez, S. Demyda-Peyras, M. Moreno, L. Alcaraz, et al. "84 EFFECT OF A STRESSOR ON CANINE SPERM DNA FRAGMENTATION USING THE SPERM CHROMATIN DISPERSION TEST." Reproduction, Fertility and Development 25, no. 1 (2013): 189. http://dx.doi.org/10.1071/rdv25n1ab84.
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Recently, a new procedure for the analysis of sperm DNA fragmentation has been developed for human and different mammalian species (Sperm-Halomax®), based on the sperm chromatin dispersion test (SCDt); however, no studies has been performed specifically on canine frozen–thawed-stressed semen but is there for cooled semen. The aim of this work was to assess the effect of a stressor (24 h in an oven at 38°C) on canine frozen–thawed semen using the SCDt to resemble what happens in the female reproductive tract. For this purpose, ejaculates were collected by digital manipulation from 4 healthy beagle dogs and the sperm-rich fraction of the ejaculates from 3 different dogs was pooled each time. All the pooled semen samples (n = 4) used presented physiological values concerning to routine semen parameters (motility, morphology, and sperm concentration). After evaluation, semen samples were centrifuged and the sperm pellet resuspended to a final concentration of 100 × 106 sperm mL–1 in 2 steps with CaniPRO Freeze (Minitub, Tiefenbach, Germany). Sperm were slowly cooled to 5°C and then loaded into 0.5-mL plastic straws. After that, straws were frozen in liquid-nitrogen vapours for 10 min and stored into a nitrogen tank. Straws were thawed in a water bath (30 s/37°C) and incubated for 24 hours at 38°C before analysis. The sperm DNA fragmentation was assessed in fresh semen and frozen–thawed-stressed samples using the Sperm-Halomax® commercial kit specifically developed for canine semen (Halotech DNA SL, Madrid, Spain) following the manufacturer’s instructions. Slides were stained for green fluorescence staining and 500 sperm per slide were counted using fluorescence microscopy. The sperm DNA fragmentation index (%) was compared between fresh and frozen–thawed-stressed semen samples by ANOVA. Results were expressed as mean ± standard error of the mean. The results obtained showed that subjecting thawed semen to 24 h in an oven at 38°C significantly increased (P < 0.05) DNA fragmentation compared with fresh semen (2.7% ± 0.2 v. 1.4 ± 0.1%). The stress factor was performed to simulate the viability of canine thawed sperm (12–24 h) when a bitch is inseminated with frozen semen. It would be interesting to perform further studies to relate sperm DNA fragmentation and fertility of frozen–thawed canine semen. In conclusion, frozen–thawed-stressed semen samples increased the sperm DNA fragmentation index measured using a SCDt. Further studies are needed to relate sperm DNA fragmentation with fertility rates or cryopreservation success.
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Anitha, Adaboina, and Burri Sandhya Rani. "Comparison of fresh and frozen thawed embryo transfer in terms of clinical pregnancy rate." International Journal of Reproduction, Contraception, Obstetrics and Gynecology 9, no. 2 (January 28, 2020): 607. http://dx.doi.org/10.18203/2320-1770.ijrcog20200345.
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Background: In a standard IVF (in-vivo fertilization) procedure, the embryos formed after the fertilization of male and female gametes are allowed to grow for 3-5 days and then transferred back to the uterine cavity of the female, where they might get attached and start to grow. Objective of this study was to compare clinical pregnancy rate of fresh embryo transfers and frozen-thawed embryo transfers.Methods: This is a retrospective case control study in patients undergoing IVF /ICSI cycles from January 2018 to December 2018 were enrolled in assisted reproduction. Total of 200 women which contains 118 fresh embryo transfers and 82 frozen-thawed embryo transfers are studied.Results: Clinical pregnancy rates of fresh cleavage-stage embryo transfers compared with frozen-thawed cleavage-stage embryo transfers, were (53.3% versus 39.6%). Ectopic pregnancy is also significant in comparison. In patients under 35 years of ages and (57.1% versus 12.5%). In patients older than 35 years old, respectively. The multiple pregnancy rates, abortion rates and ectopic pregnancy rates did not differ significantly among the groups. Multiple pregnancy rate and abortion rate is significantly high in frozen-thawed blastocyst transfer than fresh embryo transfer. Whereas the ectopic pregnancy rates had no difference in both groups.Conclusions: The clinical pregnancy rates in fresh embryo transfer is high than that of frozen-thawed blastocyst.
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Park, Sae Young, Eun Young Kim, Xiang Shun Cui, Jin Cheol Tae, Won Don Lee, Nam Hyung Kim, Se Pill Park, and Jin Ho Lim. "Increase in DNA fragmentation and apoptosis-related gene expression in frozen-thawed bovine blastocysts." Zygote 14, no. 2 (May 2006): 125–31. http://dx.doi.org/10.1017/s0967199406003649.
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SummaryEvaluation of apoptosis and expression level of apoptosis-related genes is useful for examining the variation in embryo quality according to environmental change. The objective of this study was to investigate DNA fragmentation and apoptosis-related gene expression patterns in frozen-thawed bovine blastocysts.In vitroproduced day 7 blastocysts were frozen by two different vitrification methods (conventional 0.25 ml straw or MVC straw). After thawing, DNA fragmentation of surviving embryos was examined by TUNEL assay, and the expression patterns of their apoptotic genes (survivin, Fas, Hsp 70 and caspase-3) were evaluated using real-time quantitative reverse transcriptase polymerase chain reaction.In vitrosurvival rates of frozen-thawed embryos were higher following the MVC vitrification method (88.2% re-expanded at 24 h, 77.1% hatching at 48 h) than the conventional (C) vitrification method (77.0% re-expanded at 24 h, 66.7% hatching at 48 h). However, both vitrified methods resulted in a significantly higher apoptotic index (C vitrification method 11.9%, MVC vitrification method 11.0%) than in non-frozen embryos (3.0%). Expression levels of survivin, Fas, caspase-3, and Hsp 70 were also increased in the frozen-thawed embryos compared with non-frozen embryos. These results indicate that the cryopreservation procedure might cause damage that results in an increase in DNA fragmentation and apoptosis-related gene transcription, reducing developmental capacity of frozen-thawed embryos.
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Šichtař, J., A. Nehasilová, O. Šimoník, and F. Bubeníčková. "Effect of two freezing extenders on characteristic of fresh and frozen-thawed semen in endangered Old Kladruber stallions – A pilot study." Czech Journal of Animal Science 62, No. 6 (May 24, 2017): 227–33. http://dx.doi.org/10.17221/76/2016-cjas.
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The aim of the study was to evaluate the effect of two different extenders on sperm characteristic before equilibration and post-thaw in the endangered Old Kladruber stallions. Also, the response of individual stallions to the extenders used was tested. Semen was collected from six stallions every other day within one week. After centrifugation of the collected sperm-rich fraction, the supernatant was removed and sperm pellets were divided to two aliquots; these were diluted either with Gent (Minitube, Germany) or privately manufactured lactose-EDTA-egg yolk extender (Lact). Three cryopreserved insemination doses (IDs) from each extender (Gent and Lact) were prepared for each stallion from one collection (108 samples from six stallions in total). As a parameter of quality, the motility (computer assisted sperm analysis), viability (fluorescence staining), and morphology (eosin/nigrosine staining) were evaluated after dilution with freezing extenders (fresh) and after thawing (frozen-thawed). The different effects of chosen extenders on the quality of fresh semen were only manifested in higher kinematic parameters of sperm when the Lact extender was used. However, in frozen-thawed samples, the Gent extender yielded significantly better results in all of the evaluated parameters. The representation of sperm subpopulation was significantly influenced by extender in fresh as well as frozen-thawed samples; moreover, we found a significant effect of freezing on the distribution of these subpopulations. The response of individual stallions to chosen extenders was evident in the different quality of fresh as well as frozen-thawed IDs; Gent extender yielded better frozen-thawed IDs. Based on our results, among others describing quality parameters of ejaculate in endangered Old Kladruber stallions, we can recommend using Gent extender for the production of frozen-thawed IDs.
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Jung, Y. G., T. Sakata, E. S. Lee, and Y. Fukui. "Amino acid metabolism of bovine blastocysts derived from parthenogenetically activated or in vitro fertilized oocytes." Reproduction, Fertility and Development 10, no. 3 (1998): 279. http://dx.doi.org/10.1071/r98052.
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The uptake and synthesis of 19 amino acids by fresh or frozen–thawed bovine blastocysts produced by parthenogenesis (PT) or in vitro fertilization (IVF) were compared in the present study. Fresh blastocysts, 180 h after IVF or PT activation, and frozen–thawed blastocysts, 168 h old and cultured for 12 h post-thawing, were cultured in synthetic oviduct fluid medium (SOFM) containing polyvinyl alcohol (PVA) with both essential and non-essential amino acids (EAA and NEAA, respectively) (Medium 1: M1) or SOFM containing PVA with only EAA (Medium 2: M2). In Experiment 1, when fresh or frozen–thawed PT blastocysts were cultured in M1, the uptake of glutamate (in fresh only), aspartate and arginine, and the synthesis of glutamine and alanine were significantly enhanced. In the culture with M2, serine, asparagine, glutamate, glutamine, glycine, arginine and alanine were significantly taken up. It was found that the glutamine concentrations was significantly higher (P < 0.001) in the culture medium drops containing embryos than in the drops without embryos. In Experiment 2, when PT blastocysts were cultured in M1, the uptake of aspartate and synthesis of alanine were greater (P < 0.01) than those by IVF blastocysts. When M2 was used, a significant (P < 0.01) production of serine, asparagine, glutamate, glutamine and alanine, and the uptake of arginine by PT blastocysts were observed. In Experiment 3, when IVF blastocysts were cultured in M1, fresh blastocysts depleted more aspartate and glutamate, and produced more glutamine and alanine than frozen–thawed blastocysts. When cultured in M2, frozen–thawed blastocysts depleted more threonine (P < 0.01) than fresh blastocysts. These results indicate that the uptake and synthesis of amino acids were different in fresh or frozen–thawed bovine blastocysts derived from PT or IVF. These differences in amino acid metabolism may be related to the viability of the blastocysts.
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Eftekhar, Maryam, Banafsheh Mohammadi, Nasim Tabibnejad, and Maryam Mortazavi Lahijani. "Frozen–thawed cleavage stage versus blastocyst stage embryo transfer in high responder patients." Zygote 28, no. 6 (August 27, 2020): 511–15. http://dx.doi.org/10.1017/s0967199420000428.
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SummaryClinical outcomes following frozen–thawed cleavage embryo transfer versus frozen–thawed blastocyst transfer in high responder patients undergoing in vitro fertilisation/intracytoplasmic sperm injection cycles are still debated. In a retrospective study, 106 high responder patients who were candidate for ‘freeze-all embryos’ were recruited. Frozen–thawed embryos were transferred at the cleavage stage (n = 53) or the blastocyst stage (n = 53). Clinical pregnancy was considered as the primary outcome and chemical pregnancy, ongoing pregnancy, implantation rate, and fertilization rate, as well as miscarriage rate, were measured as the secondary outcome. Clinical (47.2% vs. 24.5%), chemical (56.6% vs. 32.1%), and ongoing pregnancy rates (37.7% vs. 17%) as well as implantation rates (33.6% vs. 13.5%) were significantly higher in the blastocyst group compared with the cleavage group respectively (P < 0.05). Miscarriage rate was comparable between groups (P > 0.05). Transfer of frozen–thawed embryos at the blastocyst stage was preferable in the high responder patients to increase implantation, pregnancy and live birth rates compared with cleavage stage embryo transfer.
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Stafford-Bell, M. A., and C. M. Copeland. "Surrogacy in Australia: implantation rates have implications for embryo quality and uterine receptivity." Reproduction, Fertility and Development 13, no. 1 (2001): 99. http://dx.doi.org/10.1071/rd00044.
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Since the passage, in November 1995, of the ACT Substitute Parents Agreement Act, The Canberra Fertility Centre has added a true gestational carrier pregnancy programme to its established infertility and IVF services. Embryos generated are transferred as frozen–thawed embryos to the carrier in an average of 2.2 embryos per transfer. Between 1 January 1996 and 31 December 1999 the results of 49 frozen embryo transfers to 25 gestational carriers were compared with 849 frozen embryo transfers on a routine IVF programme. In the carrier group, the embryo implantation rate of 13.8% per embryo transferred is double that of an exactly comparable group of patients undergoing routine frozen–thawed embryo transfer on the same IVF programme and considerably higher than those reported in large series of frozen–thawed embryo transfers. Exclusion from the carrier pregnancy programme of patients with incipient ovarian failure results in an implantation rate of 16.7%, a clinical pregnancy rate of 29.0% and a live birth rate of 19.4% per embryo transfer procedure.
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Yasri, Rakhmi, Ristika Handarini, and Muhammad Imron. "THE QUALITY OF EMBRYOS RESULTED FROM IN VITRO FERTILIZATION BY USING FROZEN SEMEN THAWED IN DIFFERENT TEMPERATURES." Jurnal Peternakan Nusantara 3, no. 1 (October 13, 2017): 23. http://dx.doi.org/10.30997/jpnu.v3i1.853.
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In vitro fertilization technology in cows is an effort done to utilize ovary waste from cows slughtered in abbatoir. This study was aimed at assessing the qualiy of embryos resulted from in vitro fertilization by using frozen semen thawed in different temperatures. In order to get qualty semen, standardized thawing method is required. It was expected from this study that an optimum thawing temperature for frozen semen was determined to obtain quality transferable embryos. Three treatments consisting of thawing with water 37°C for 30 second (T1), thawing with water 25°C for 30 second (T2), and thawing with water 10°C for 30 second (T3). Data were subjected to an an anlysis of variance (Anova) and a Duncan test. Results showed that oocytes fertilized with frozen semen thawed at 37°C and 10°C had higher fertilization rate and excellent-grade embryos (P<0.05) than did the ones fertilized with frozen semen thawed at 25°C. However, no different effect of thawing temperatures was found on transferable and degenerated embryos (P>0.05). It was concluded that embryos fertilized with Brahman frozen semen in thawed at 37°C had the highest number of embryos (49.66±2.88) and excellent-grade embryos (22.00±4.35). Key words: Embryo quality, In vitro fertilization, frozen semen thawing, Brahman bull.
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Aleksyutina, D. M., and R. G. Motenko. "Effect of soil salinity and organic matter content on thermal properties and unfrozen water content of frozen soils at west coast of Baydara Bay." Moscow University Bulletin. Series 4. Geology, no. 2 (April 28, 2016): 59–63. http://dx.doi.org/10.33623/0579-9406-2016-2-59-63.
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The paper considers laboratory results of the composition, structure and properties for frozen and thawed soils at west coast of Baydara Bay. Experimental data of unfrozen water content and thermal properties are discussed and summarized for different soils. The roles of soil salinity and organic matter content on these data were estimated for frozen and thawed soils.