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1.
Anim Reprod Sci ; 226: 106688, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33476904

RESUMEN

Effects of amount of anechoic intrauterine fluid (IUF) and size of largest follicle at insemination on proportion of cows pregnant as a result of AI (P/AI percentage), as well as factors associated with amount of IUF and size of the follicle were investigated. Follicle size was determined, and amount of IUF in the largest uterine segment was categorized as containing no fluid, ≤10, >10-20, and >20 mm using the ultrasonic monitor grid in 735 cows at time of insemination. Multivariable regression models were constructed to evaluate effects of different variables on P/AI percentage, and variables associated with the IUF score and follicle size. On day 30 post-insemination, the P/AI percentage was greater in cows that had >10 to 20 mm IUF than cows with no IUF (OR = 1.9, P = 0.01), but on day 70 post-insemination, the P/AI percentage was similar in cows with different amounts of IUF. Follicle size was not associated with P/AI percentage on days 30 and 70 post-insemination. Cows in spontaneous estrus, multiparous cows, cows with a cystic structure and a typically functional follicle, and Holstein cows had a greater likelihood for a larger IUF value. Primiparous cows, cows on which there was imposing of a hormonal-ovulation-synchronization regimen, and crossbred cows were more likely to have smaller follicles. In conclusion, amount of IUF at the time of insemination and size of the follicle were not associated with P/AI percentage, and several variables were associated with amount of IUF or follicle size.


Asunto(s)
Líquidos Corporales/diagnóstico por imagen , Bovinos/fisiología , Inseminación Artificial/veterinaria , Animales , Estudios de Cohortes , Femenino , Embarazo , Estudios Prospectivos , Ultrasonografía/veterinaria , Útero
2.
Int J Urol ; 28(3): 333-338, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33327043

RESUMEN

OBJECTIVE: To evaluate the safety, efficacy and cost-effectiveness of holmium enucleation of the prostate and bipolar transurethral enucleation of the prostate. METHODS: In our randomized controlled trial, 120 patients were allocated into two equal groups representing holmium enucleation of the prostate and bipolar enucleation of the prostate. Operative parameters were recorded according to operative, enucleation and resection time in addition to the intraoperative complications. Patients were followed up at 1, 3 and 12 months postoperative to assess the prostate size, post-voiding residual urine, International Prostate Symptom Score, peak urine flow rate and quality of life, and compared with the preoperative parameters. Cost analysis was evaluated for both procedures. RESULTS: We evaluated 107 patients who finished our follow up and their data were analyzed. The prostate size was 135.2 ± 34.8 mL and 125 ± 26.9 mL for holmium enucleation of the prostate and bipolar enucleation of the prostate, respectively. Holmium enucleation of the prostate was associated with a shorter operative time of 83.43 ± 6.92 min compared with 94.7 ± 12.2 min in bipolar enucleation of the prostate groups. Holmium enucleation of the prostate was associated with an earlier catheter removal time and shorter hospital stay compared with bipolar enucleation of the prostate. Postoperative International Prostate Symptom Score, quality of life, post-voiding residual urine, peak urine flow rate, prostate-specific antigen and prostate volume reduction were comparable between both groups, and they both showed statistically significant improvement compared with their preoperative parameters. In the cost analysis, holmium enucleation of the prostate was more cost-effective than bipolar enucleation of the prostate. CONCLUSION: Both holmium enucleation of the prostate and bipolar enucleation of the prostate are safe and effective in the surgical management of large prostatic adenomas. Holmium enucleation of the prostate has a shorter operative time and hospital stay with earlier catheter removal time, and is more cost-effective than bipolar enucleation of the prostate.


Asunto(s)
Terapia por Láser , Láseres de Estado Sólido , Hiperplasia Prostática , Resección Transuretral de la Próstata , Humanos , Láseres de Estado Sólido/uso terapéutico , Masculino , Hiperplasia Prostática/cirugía , Calidad de Vida , Resección Transuretral de la Próstata/efectos adversos
3.
Theriogenology ; 154: 171-180, 2020 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-32622197

RESUMEN

The efficacy of five ovulation-synchronization protocols with FTAI in treatment of cows diagnosed with follicular cysts was investigated in a nonrandomized study in a single herd. Cows identified with follicular structures >25 mm on two subsequent ultrasonographic (USG) examinations 7-10 days apart (n = 552) were assigned to one of the five treatment regimens on the day of the second USG examination. Treatment regimens were Ovsynch (GnRH-7d-PGF2α-56 h-GnRH-16 h-FTAI), New-CIDR (as Ovsynch with a new CIDR insert being fitted between days 0 and 7), Reused-CIDR (as New-CIDR but the CIDR insert was previously used for 7 days in another cow), G-New CIDR (Pre-GnRH on day 0 with the New-CIDR regimen being initiated 7 days later), and RG-Ovsynch (Pre-GnRH on day 0 and then every 7 days until detection of a luteal structure upon USG examination; at that point the Ovsynch was initiated). A subset of cows was subjected to ovarian USG examination at the time of PGF2α administration, at insemination, and 8-10 days post-insemination. Progesterone-releasing ability of new and reused CIDR inserts were evaluated in cows diagnosed with severe ovarian inactivity (n = 16). The data were analyzed using logistic regression with pregnancy per AI on days 30 (P/Al 30) and 70 (P/AI 70) post-insemination were included as outcome measures. Compared with Ovsynch, RG-Ovsynch improved the P/AI 30 (OR = 2.6, P = 0.03) and the P/AI 70 (OR = 2.5, P = 0.05). New-CIDR and G-New CIDR were associated with non-significant increase in P/AI 30 (OR = 2.1, P = 0.09 and OR = 2.3, P = 0.07, respectively) and P/AI 70 (OR = 2.01, P = 0.09 and OR = 2.2, P = 0.09, respectively). Reused-CIDR was not associated with improvement in P/AI (P = 0.93 and 0.79 for P/AI 30 and P/AI 70, respectively). RG-Ovsynch had a longer diagnosis-to-FTAI interval (median 24, IQR 17,31). The dominant ovarian structures and the presence of a cyst or a luteal structure at PGF2α administration or at insemination were not associated with P/AI. The new and re-used CIDR inserts produced comparable concentrations of serum progesterone 3 h, 3 days and 7 days post CIDR insertion. In conclusion, the RG-Ovsynch improved the P/AI whereas the New-CIDR and the G-New CIDR regimens tended to increase the P/AI compared with Ovsynch. Marginal differences in P/AI between RG-Ovsynch, New-CIDR, and G-New-CIDR together with prolonged diagnosis-to-FTAI in RG-Ovsynch should be considered if to evaluate the economic value of these regimens.


Asunto(s)
Enfermedades de los Bovinos , Quiste Folicular , Animales , Bovinos , Dinoprost/farmacología , Sincronización del Estro , Femenino , Quiste Folicular/veterinaria , Hormona Liberadora de Gonadotropina , Inseminación Artificial/veterinaria , Ovulación , Embarazo , Progesterona
4.
Anim Reprod Sci ; 216: 106356, 2020 May.
Artículo en Inglés | MEDLINE | ID: mdl-32414465

RESUMEN

In this study, there was evaluation of pregnancy per AI (P/AI) as a result of the first postpartum AI following four ovulation-synchronization treatments. Treatment regimens were Ovsynch-56 (OVS, n = 875; GnRH1-7d-PGF2α-56h-GnRH2-16h-FTAI), CIDR-OVS (n = 1001; OVS plus CIDR inserts between GnRH1 and PGF2α), Double-Ovsynch (DOVS, n = 663; imposing Pre-OVS followed by breeding-OVS 7 days later), and Modified-DOVS (M-DOVS, n = 1397; imposing Pre-OVS followed either by breeding-OVS or CIDR-OVS based upon the ovarian structure at GnRH1). Cows with a corpus luteum were assigned to a breeding-OVS treatment regimen and those that did not have a corpus luteum were assigned to the CIDR-OVS treatment regimen. Compared with OVS, the P/AI of the cows in the M-DOVS (OR = 1.5, P = 0.001) and CIDR-OVS (OR = 1.4, P = 0.017) was greater at day 30. At day 70, only in the M-DOVS group was there a greater P/AI compared with the OVS group (OR = 1.7, P < 0.001). Pregnancy loss between days 30 and 70 was greater in cows of the CIDR-OVS (OR = 1.9, P = 0.014) compared with those of the OVS group. In cows of the M-DOVS, the dominant ovarian structures (follicle, corpus luteum or cyst) at different time-points of the pre-synchronization period and occurrence of estrus at the end of this period were not associated with P/AI at day 30 post-AI. In conclusion, imposing CIDR-OVS in cows that did not respond to pre-synchronization treatments, resulted in an enhanced pregnancy percentage with the use of the DOVS.


Asunto(s)
Bovinos , Dinoprost/farmacología , Sincronización del Estro/métodos , Hormona Liberadora de Gonadotropina/farmacología , Inseminación Artificial/veterinaria , Animales , Dinoprost/administración & dosificación , Femenino , Hormona Liberadora de Gonadotropina/administración & dosificación , Embarazo , Índice de Embarazo
5.
Animals (Basel) ; 9(11)2019 Nov 19.
Artículo en Inglés | MEDLINE | ID: mdl-31752406

RESUMEN

This study compared the efficiency of commercial egg yolk-free (AndroMed, OPTIXcell) and egg yolk-supplemented (Triladyl, Steridyl) Tris-based extenders for semen cryopreservation in seven adult dromedary camels. The camel-specific extender SHOTOR was used as control. The collected semen samples were evaluated and diluted with SHOTOR, Triladyl, Steridyl, AndroMed, or OPTIXcell. The diluted semen was gradually cooled and equilibrated for two hours before liquid nitrogen freezing. Semen was evaluated prior to freezing and after freeze-thawing cycles for motility, kinetics, vitality, abnormality, plasma membrane integrity, and DNA fragmentation using computer-assisted sperm analysis. In pre-freezing evaluation, progressive sperm motility was higher in SHOTOR-diluted samples (21.54 ± 1.83) than in samples diluted with Steridyl, OPTIXcell, or AndroMed (15.76 ± 1.80, 17.43 ± 1.10, and 13.27 ± 1.07, respectively). Moreover, Triladyl and SHOTOR resulted in significantly (p < 0.05) better sperm vitality and DNA integrity than all other diluents, but Triladyl resulted in a significantly (p < 0.05) better plasma membrane integrity (87.77 ± 0.31) than SHOTOR (85.48 ± 0.58). In the post-thawing evaluation, Triladyl led to significantly (p < 0.05) higher sperm motility (38.63 ± 0.81%; p < 0.05) when compared to SHOTOR, Steridyl or AndroMed (35.09 ± 1.341%, 34.4 ± 0.84%, and 31.99 ± 1.48%, respectively), with OPTIXcell being the least efficient (28.39 ± 0.86%). Progressive sperm motility was the highest when using Triladyl. Post-thawing curvilinear, straight line and average path sperm velocities were highest with Triladyl and lowest with AndroMed. Triladyl led to the highest linearity coefficient and straightness sperm coefficient, while SHOTOR to the highest DNA and plasma membrane integrity. OPTIXcell and AndroMed resulted in poor post-thawing sperm vitality, while Steridyl was less efficient than Triladyl. The highest rate of sperm abnormalities was recorded with OPTIXcell and the lowest with SHOTOR or Triladyl. In conclusion, SHOTOR, Triladyl, Steridyl, AndroMed, and OPTIXcell can all be used for camel semen cryopreservation; however, SHOTOR and Triladyl provided the best post-thawing sperm quality. Based on our findings, Triladyl is the best commercially available extender for dromedary camel semen cryopreservation to date.

6.
Anim Reprod Sci ; 209: 106147, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31514918

RESUMEN

In this single herd observational study, there is investigation of the effects of 81 sires (with 11,424 artificial inseminations) on conception rates in 1790 Holstein cows for 5 years. Sires were catagorized based on the published sire conception rate (SCR) into different sire fertility groups (low, average and high fertility sires). The performance of different-sire fertility groups was assessed in timed artificial insemination (TAI) and repeat-breeder (RB) cows. With this aim, two logistic regression models with sire, inseminator, cow, and lactation random effects were applied to data on pregnancies assessed at days 30 and 70 post-insemination. Fixed effects of sire fertility group, sire breed, cow-fertility status, insemination type, postpartum problems, milk yield, temperature humidity index, and year were evaluated. Results from the analysis indicated there was a significant individual sire effect on conception rates, and large heterogeneity in values for this variable among sires. Results indicate that SCR could be assessed to predict low fertility sires reasonably well, and the predicted probabilities for pregnancy per AI (P/AI) at 30 and 70 days post-insemination for high fertility sires were consistent for the most part with values for the SCR. The sire breed did not affect conception rates at days 30 and 70 post-insemination nor its interactions with insemination type (estrous detection AI (EDAI) compared with TAI) and cow-fertility status (RB compared with non-RB). Predicting response probabilities for sires with at least 100 inseminations in each insemination group resulted in greater conception probabilities in cows in which there was EDAI than those in the TAI group.


Asunto(s)
Cruzamiento , Bovinos , Padre , Fertilidad/fisiología , Inseminación Artificial , Índice de Embarazo , Animales , Cruzamiento/métodos , Cruzamiento/estadística & datos numéricos , Industria Lechera/métodos , Sincronización del Estro/fisiología , Femenino , Fertilización/fisiología , Inseminación Artificial/métodos , Inseminación Artificial/estadística & datos numéricos , Inseminación Artificial/veterinaria , Masculino , Embarazo
7.
Andrologia ; 51(3): e13214, 2019 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-30488469

RESUMEN

This study was carried out to investigate the potential effects of Chlorella vulgaris (CV) on testicular function and oxidant/antioxidant status in normal and deltamethrin-intoxicated rats. Forty adult male rats were drenched either with normal saline, CV (50 mg/kg), deltamethrin (DM) (3 mg/kg), or CV combined with DM, daily for 8 weeks. At the end of the protocol, the epididymal sperm quality was evaluated and the testicular superoxide dismutase (SOD), catalase enzyme (CAT) and malondialdehyde (MDA), and the serum testosterone levels were estimated. Normal rats treated with CV showed a significant increase in the total sperm number/epididymal tail, testicular SOD, and CAT levels with a significant decrease in the testicular MDA. Deltamethrin intoxication significantly decreased the proportions of motile and live sperm, the testosterone concentration, the testicular SOD and CAT levels, whereas it significantly increased the proportion of abnormal sperm and the testicular MDA. Chlorella vulgaris treatment significantly ameliorated the adverse effects of DM-intoxication and restored most of the parameters to levels that are comparable to those of the control group. In conclusion, CV administration improved the testicular function of normal rats and ameliorated the effect of severe oxidative stress conditions.


Asunto(s)
Antioxidantes/farmacología , Chlorella vulgaris , Nitrilos/toxicidad , Estrés Oxidativo/efectos de los fármacos , Extractos Vegetales/farmacología , Piretrinas/toxicidad , Espermatozoides/efectos de los fármacos , Testículo/efectos de los fármacos , Animales , Catalasa/metabolismo , Epidídimo/efectos de los fármacos , Epidídimo/metabolismo , Glutatión/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Masculino , Malondialdehído/metabolismo , Ratas , Motilidad Espermática/efectos de los fármacos , Espermatozoides/metabolismo , Superóxido Dismutasa/metabolismo , Testículo/metabolismo
8.
World J Urol ; 36(7): 1139-1147, 2018 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-29450731

RESUMEN

PURPOSE: The extracorporeal shockwave lithotripsy (ESWL) remains the most common first line of treatment for renal stones in the pediatric population. The purpose of this study is to evaluate and compare the outcomes of the ESWL and mini-percutaneous nephrolithotomy (mini-PCNL). PATIENTS AND METHODS: A total of 108 patients younger than 12 years of age with 1-2 cm single renal stone (pelvic or calyceal) were randomized into two groups, each containing 54 patients. Patients in group A were subjected to mini-PCNL using 16.5 Fr percutaneous sheath while those in group B underwent ESWL using Dornier Compact Sigma. RESULTS: The stone-free rate (SFR) after first session was 88.9% (48 cases) and 55.6% (30 cases) for groups A and B, respectively. The difference is highly statistically significant P = 0.006. Two patients (3.7%) in group A needed 2nd session of PCNL, while 18 patients (33.3%)in group B needed a 2nd session, of theses 18 patients six patients needed a 3rd session of ESWL. After the third session of ESWL and second look PCNL the stone-free rates were 92.59% (50 cases) and 88.89% (48 cases) for groups A and B, respectively, (P = 0.639), which is statistically insignificant. The mean hospital stay and fluoroscopy exposure were significantly longer in the mini-PCNL group. The complication rate in groups A and group B were (22.2%) and (14.8%), respectively, which is statistically insignificant (P = 0.484). CONCLUSIONS: According to Clavien grade of complications mini-PCNL is a safe procedure, and after three session of ESWL, mini-PCNL has a similar stone-free rate with a lower retreatment rate. However, the mini-PCNL has more radiation exposure, and requires a longer hospital stay.


Asunto(s)
Tratamiento con Ondas de Choque Extracorpóreas/métodos , Cálculos Renales/terapia , Nefrolitotomía Percutánea/métodos , Niño , Preescolar , Tratamiento con Ondas de Choque Extracorpóreas/efectos adversos , Femenino , Humanos , Masculino , Nefrolitotomía Percutánea/efectos adversos , Estudios Prospectivos , Resultado del Tratamiento
9.
Theriogenology ; 100: 16-23, 2017 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-28708529

RESUMEN

Defining factors associated with embryonic and/or fetal losses will be helpful in overcoming such problem, either by adjusting conditions or applying therapeutic approaches to high-risk cows. The objective of this study was to investigate the association between a number of maternal and non-maternal factors and the risk of late embryonic (LED) and early fetal death (EFD) in dairy herds. Additionally, we investigated the effect of treating pregnant cows either with GnRH on day 26 post-insemination, or GnRH on day 26 plus CIDR insertion between days 26 and 33 post-insemination, on the risk of LED/EFD. From 3826 pregnancies, diagnosed at day 30 post-insemination, 851 cows lost the pregnancy by day 70 post-insemination. A mixed-effects logistic model was constructed to assess the effect of cow breed, calving difficulty, postpartum problems, lactation number, days in milk, insemination number, actual 305-day milk production, temperature humidity index (THI) at insemination, estrus synchronization protocols, and other factors, on the risk of LED/EFD. Our findings indicated that Holstein X Brown Swiss crossbreed cows had a lower risk for LED/EFD than Holstein cows (P < 0.05). Cows that had postpartum problems, were inseminated for the first time, produced more milk, or were inseminated at THI ≥75, recorded higher risks of LED/EFD (P < 0.05). Calving difficulty, lactation number, and synchronization protocols were not found to be associated with LED/EFD. Moreover, treatment of the pregnant cows with GnRH on day 26 post-insemination plus CIDR insertion between days 26 and 33 post-insemination decreased the risk of LED/EFD. In conclusion, cows that had postpartum problems, were inseminated early postpartum, produced higher milk, and/or were inseminated at high THI, were under higher risk of LED/EFD. Treating such cows with GnRH on day 26 plus CIDR insertion between days 26 and 33 may decrease the possibility of the LED/EFD.


Asunto(s)
Aborto Veterinario/etiología , Enfermedades de los Bovinos/etiología , Animales , Bovinos , Femenino , Humedad , Lactancia , Modelos Biológicos , Periodo Posparto , Embarazo , Temperatura
10.
J Reprod Dev ; 57(5): 557-63, 2011 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22052044

RESUMEN

Freeze-drying (lyophilization) has been proposed as an alternative method for sperm preservation to overcome the disadvantages of the current cryopreservation method such as the high maintenance cost of frozen stocks, the problems associated with transportation of frozen materials and the potential risk of total loss of the frozen stock. Since freeze-dried spermatozoa after rehydration lose their motility, which is an essential requirement to complete physiological fertilization, a relatively difficult microinsemination technique must be applied to rehydrated spermatozoa. Theoretically, it has been supposed that freeze-dried spermatozoa could maintain their functions and abilities to interact with the oocyte cytoplasm after prolonged storage at refrigerator temperature. However, sufficient yield of transferable blastocysts and production of live offspring derived from freeze-dried sperm samples are still subjects to be challenged and overcome in large domestic species.


Asunto(s)
Criopreservación/métodos , Preservación de Semen/métodos , Espermatozoides , Animales , Criopreservación/normas , Criopreservación/veterinaria , Liofilización/métodos , Liofilización/veterinaria , Humanos , Masculino , Modelos Biológicos , Técnicas Reproductivas Asistidas/normas , Técnicas Reproductivas Asistidas/tendencias , Técnicas Reproductivas Asistidas/veterinaria , Preservación de Semen/efectos adversos , Preservación de Semen/normas , Preservación de Semen/tendencias , Inyecciones de Esperma Intracitoplasmáticas/métodos , Inyecciones de Esperma Intracitoplasmáticas/tendencias , Inyecciones de Esperma Intracitoplasmáticas/veterinaria
11.
J Reprod Dev ; 57(3): 428-32, 2011 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-21325738

RESUMEN

This study was designed to investigate whether freeze-dried (FD) bull spermatozoa maintained the function of the microtubule-organizing center (MTOC) after rehydration and intracytoplasmic sperm injection (ICSI). In a preliminary attempt, the cleavage and blastocyst formation rates in FD-ICSI zygotes (36 and 1%, respectively) were found to be considerably lower than those in control ICSI zygotes (67 and 21%, respectively) or in IVF zygotes (78 and 43%, respectively). An alkaline comet assay indicated that the DNA fragmentation index (length of comet tail % DNA liberated) was not significantly different between fresh and FD spermatozoa. In the main experiment, formation of sperm-asters in the FD-ICSI oocytes 7 h postinsemination occurred at a similar rate when compared with the control ICSI oocytes (41 vs. 49%). Among the oocytes exhibiting sperm aster formation, the extent of microtubule network assembly was comparable between the FD-ICSI and control ICSI groups. However, the MTOC of the ICSI oocytes was not as functional as that of IVF oocytes in terms of the aster formation rate (97%) and the fluorescent intensity of the microtubule network (2.0 folds). These results suggest that the freeze-drying process per se had no adverse effect on maintaining the MTOC function in bull spermatozoa.


Asunto(s)
Centro Organizador de los Microtúbulos/fisiología , Preservación de Semen/métodos , Inyecciones de Esperma Intracitoplasmáticas/efectos adversos , Espermatozoides/fisiología , Animales , Blastocisto , Bovinos , Fragmentación del ADN , Desarrollo Embrionario , Liofilización , Masculino
12.
J Reprod Dev ; 55(4): 356-60, 2009 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-19721335

RESUMEN

Epigenetic reprogramming in early preimplantation embryos, that refers to erasing and remodeling epigenetic marks such as DNA methylation, is essential for differentiation and development. In many species, paternal genome is subjected to genome-wide active demethylation before the DNA replication commences, while maternal genome maintains its methylation status until being demethylated passively during the subsequent cleavage divisions. The purpose of this manuscript was to review the available knowledge about the paternal genome active demethylation process concerning the possible mechanisms, species variation and the factors affecting the active demethylation dynamics such as in vitro protocols for production of pronuclear-stage zygotes. Better understanding the mechanisms by which the epigenetic reprogramming is occurred may contribute to clarify the biological significance of this process.


Asunto(s)
Genoma , Cigoto/metabolismo , Animales , Metilación de ADN , Desarrollo Embrionario , Epigénesis Genética , Femenino , Fertilización , Regulación de la Expresión Génica , Humanos , Cinética , Masculino , Modelos Biológicos , Especificidad de la Especie
13.
J Reprod Dev ; 55(4): 433-9, 2009 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-19403998

RESUMEN

This study was designed to investigate the dynamics of the paternal genome demethylation in pronuclear-stage bovine zygotes produced either by in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) using freeze-thawed (FT) as well as freeze-dried (FD) bull sperm stored at +4 or -196 C for one year. Zygotes were fixed and immunostained using anti-5-methyl-cytosin at 8, 10, 14 and 18 h post IVF (hpi) and at 6 and 12 h post ICSI (hpic). In conventional IVF-derived zygotes, the overall average of the relative methylation (RM; male/female) decreased from 0.92 at 8 hpi to 0.69 at 10 hpi (P<0.05) without any additional decrease at 14 and 18 hpi (0.67 and 0.64, respectively; P>0.05). This was accompanied by higher proportions of zygotes showing RM<0.6 (45.5, 37.5 and 38.2% at 10, 14 and 18 hpi, respectively; P<0.05) compared with 3.7% at 8 hpi. The overall averages of the RM in the FT-ICSI derived zygotes (0.79 and 0.66 at 6 and 12 hpic, respectively) were similar to those in the corresponding IVF-derived zygotes (8 and 14 hpi), but a higher proportion of the 6 hpic zygotes (37.8%; P<0.05) showed an RM<0.6 compared with the 8 hpi zygotes (3.7%). The proportions of FD-ICSI derived zygotes at 12 hpic showing an RM<0.6 (60.6 and 62.4% for +4 and -196 C storage, respectively) were higher than that of the FT-ICSI derived zygotes (39.4%; P<0.05). Thus, the bovine paternal genome rapidly demethylated within 10 h after IVF and 6 h after ICSI, and the freeze-drying and/or the storage process had no adverse effect on demethylation of the paternal genome. The extent of demethylation in the pronuclear-stage bovine zygotes was moderate, with 0.4< or =RM<0.6.


Asunto(s)
Fertilización In Vitro/métodos , Oocitos/metabolismo , Espermatozoides/metabolismo , Cigoto/metabolismo , 5-Metilcitosina/farmacología , Animales , Bovinos , Femenino , Liofilización , Genoma , Masculino , Metilación , Modelos Biológicos , Modelos Estadísticos , Inyecciones de Esperma Intracitoplasmáticas/métodos , Factores de Tiempo
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