Preservation and artificial insemination of sexed semen in sheep

2004 ◽  
Vol 16 (4) ◽  
pp. 455 ◽  
Author(s):  
G. Evans ◽  
F. K. Hollinshead ◽  
W. M. C. Maxwell
Keyword(s):  
Artificial Insemination ◽  
In Vitro Fertilisation ◽  
Sperm Viability ◽  
Multiple Ovulation ◽  
Sexed Semen ◽  
Before And After ◽  
Advanced Stages ◽  
Sorting Process ◽  
And Function

Sperm-sexing technology using flow cytometry is in advanced stages of development for the sperm of several species. The sorting process could compromise sperm viability and sperm require specific handling procedures both before and after sorting to maintain the integrity and function of the sorted sperm. Standard freezing protocols have been modified for post-sorting cryopreservation of sperm and frozen sperm have been successfully thawed, sorted, refrozen and subsequently used to produce offspring. The relatively low numbers of available sorted sperm have, in some cases, led to modification of artificial insemination techniques to maximise efficiency of use. Multiple ovulation and embryo transfer, or in vitro fertilisation and associated technology, may lead to the more efficient use of sexed sperm.

Assessment of sperm quality parameters of six bulls showing different abilities to promote embryo development in vitro

2006 ◽  
Vol 18 (3) ◽  
pp. 395 ◽  
Author(s):  
M. Alomar ◽  
J. Mahieu ◽  
B. Verhaeghe ◽  
L. Defoin ◽  
I. Donnay
Keyword(s):  
Sperm Quality ◽  
Sharp Decrease ◽  
Quality Parameters ◽  
In Vitro Fertilisation ◽  
Sperm Viability ◽  
Fertilisation Rate ◽  
Significant Difference ◽  
Before And After ◽  
Development In Vitro

Experiments were conducted to investigate the possible origins of variation between six bulls showing various blastocyst rates after in vitro fertilisation. No significant difference was observed for the rates of cleavage and 5–8 cell stages, whereas blastocyst yields at Day 6, 7 and 8 post insemination were significantly different between bulls (P < 0.05). Fertilisation rates ranged from 59.5 to 79.3% (P < 0.05), with no difference in the incidence of polyspermy. The proportions of motile and progressive spermatozoa before and after Percoll separation were analysed. A positive effect of Percoll was noted on both parameters (P < 0.05), leading to the absence of difference between bulls after the separation process. Sperm viability and spontaneous acrosome reaction were assessed during 18 h incubation in fertilisation medium. A sharp decrease in sperm viability was observed for all bulls after 2 h incubation, with only 12.6–21.7% of spermatozoa still viable at 18 h. In contrast, the proportion of reacted acrosomes was low in five out of six bulls (<15% at 18 h). In conclusion, the fertilisation rate was the only parameter to show some correlation with blastocyst rate for all bulls.

In vitro fertilisation of mouse oocytes reconstructed by transfer of metaphase II chromosomes results in live births

Zygote ◽  
2001 ◽  
Vol 9 (1) ◽  
pp. 9-14 ◽  
Author(s):  
Min-Kang Wang ◽  
Da-Yuan Chen ◽  
Ji-Long Lui ◽  
Guang-Peng Li ◽  
Qing-Yuan Sun
Keyword(s):  
Nuclear Transfer ◽  
Normal Structure ◽  
Human Reproduction ◽  
In Vitro Fertilisation ◽  
Kunming Mouse ◽  
Mouse Oocytes ◽  
Assisted Human Reproduction ◽  
And Function ◽  
Apparatus Transfer

The interaction between nucleus and cytoplasm can be explored through nuclear transfer. We describe here another tool to investigate this interaction: MII meiotic apparatus transfer (MAT) between mouse oocytes. In this study, the MII oocyte meiotic apparatus or spindle from C57BL/6 mice, a black strain, was transferred into an enucleated metaphase oocyte from Kunming mouse, a white strain. The results showed that the enucleation rate by treating oocytes with 3% sucrose was 100%, but the electrofusion efficiency was very low, with only 17.6% of reconstructed karyoplast-recipient cytoplasm pairs fused. When the fused oocytes were exposed to spermatozoa from C57BL/6 mice, 9 of 11 (82%) were fertilised. Eight reconstructed embryos at 1- to 4-cell stages were transferred into the oviducts of two synchronously pregnant Kunming strain fosters and one delivered two normal C57BL/6 offspring. This study indicates that MII meiotic apparatus or spindle sustains normal structure and function after micromanipulation and electrofusion. MAT provides a model for further research on the application of this technique to assisted human reproduction.

Sexual activity and function in women with advanced stages of pelvic organ prolapse, before and after laparoscopic or vaginal mesh surgery

2020 ◽  
Author(s):  
Sònia Anglès-Acedo ◽  
Cristina Ros-Cerro ◽  
Sílvia Escura-Sancho ◽  
M. José Palau-Pascual ◽  
Eduardo Bataller-Sánchez ◽  
...  
Keyword(s):  
Pelvic Organ Prolapse ◽  
Sexual Activity ◽  
Pelvic Organ ◽  
Vaginal Mesh ◽  
Mesh Surgery ◽  
Before And After ◽  
Advanced Stages ◽  
And Function

92 EVALUATION OF BOAR SPERM FUNCTIONALITY AFTER A CUSHIONED CENTRIFUGATION TECHNIQUE

2006 ◽  
Vol 18 (2) ◽  
pp. 154
Author(s):  
J. Gadea ◽  
S. Martínez-Miró ◽  
G. Decuadro-Hansen ◽  
C. Matás
Keyword(s):  
Seminal Plasma ◽  
Acrosome Reaction ◽  
Sperm Viability ◽  
Lipid Packing ◽  
Lipid Disorder ◽  
Merocyanine 540 ◽  
Centrifuge Tube ◽  
Before And After ◽  
Centrifugation Technique

Separation of sperm from seminal plasma is required in most semen freezing procedures. Semen is typically subjected to centrifugation to concentrate sperm into a pellet and allow removal of the seminal plasma prior to dilution in freezing extender. Centrifugation is a relatively effective method to recover sperm, however, the process also causes considerable sperm damage. The use of a dense, inert, and isotonic solution as a cushion in the bottom of the centrifuge tube allows a greater centrifugation speed to be applied and results in greater sperm recovery. The aim of the present work was to evaluate the effects of this cushioned centrifugation technique on in vitro sperm viability and functionality. Sperm-rich fractions from 16 fertile boars were diluted and cooled to 15�C; then subsamples were centrifuged by one of two different techniques. A standard method (SM), 800 g for 10 min in 50-mL tubes (Westendorf et al. 1975 Dtsch. Tier�rztl. Wschr. 82, 261-267) and a cushioned method (CM), 1000 g for 20 min using 45 mL of diluted semen on 5 mL of an isotonic iodixanol solution (60% w/v gradient) were performed. Sperm samples were stained with merocyanine 540 (M540) and Yo-Pro 1 (Harrison et al. 1996 Mol. Rep. Dev. 45, 378-391) to detect changes in lipid packing disorder of the plasma membrane. Another set of sperm samples was incubated in the presence of (0.7 �M) 22,72-dichlorodihydrofluorescein diacetate (Gadea et al. 2005 J. Androl. 26, 396-404) to estimate production of reactive oxygen species (ROS). A final set of sperm samples was stained with peanut aggultinin-fluorscein isothiocyanate (PNA-FITC) and propidium iodide to evaluate the acrosome reaction. All of these parameters were evaluated by flow cytometry before and after centrifugation. ANOVA analysis revealed that centrifugation altered lipid packing disorder and viability. Raw semen (RS) had a larger number of viable low lipid disorder sperm than centrifuged semen (RS = 86.9a vs. SM = 81.64b vs. CM = 80.6b, P < 0.01) and a decreased number of dead sperm cells (RS = 9.5a vs. SM = 15.0b vs. CM = 16.3b, P < 0.01). However, the cushioned and standard centrifugation methods yielded similar results for all the parameters measured. No significant differences were found for generation of ROS or in the number of sperm exhibiting the acrosome reaction. In conclusion, compared to the standard centrifugation method, this simple cushioned modification is a more efficient means of processing boar semen for freezing because significantly less sample losses are detected; also, it provides similar levels of sperm viability and functionality, and consequently a higher number of doses per ejaculation can be produced.

scholarly journals Sodium bicarbonate and HEPES as buffer components for cooling the semen of pony stallions

2018 ◽  
Vol 39 (2) ◽  
pp. 631
Author(s):  
Janislene Mach Trentin ◽  
Luiz Augusto Machado Centeno ◽  
Taison De Souza Balestrin ◽  
Thainá Minela ◽  
Guilherme Machado Zanatta ◽  
...  
Keyword(s):  
Sodium Bicarbonate ◽  
Artificial Insemination ◽  
Membrane Integrity ◽  
Milk Powder ◽  
Skim Milk ◽  
Skim Milk Powder ◽  
Mitochondrial Activity ◽  
Sperm Viability ◽  
Progressive Motility ◽  
Before And After

The composition of semen diluents can modify its viability during cooling. The buffering effects of HEPES and sodium bicarbonate were evaluated considering the pH and sperm viability. The semen of seven adult Brazilian ponies was evaluated before and after cooling at 5oC for 24 h and 48 h. A non-buffered skim milk powder extender (C) and the same extender buffered with sodium bicarbonate (SB) and HEPES (H) were used. After dilution, semen (three ejaculates/pony) was centrifuged and the seminal plasma discarded. Sperm was then diluted with SB, H or C and its concentration adjusted to 50 x 106 sptz/mL. Progressive motility evaluated after dilution showed similar results with all extenders (71.42% (SB), 74.28% (H), and 74.52% (C)). Sperm motility was evaluated 24 h and 48 h after cooling for SB (44.76% and 25.23%), H (51.42% and 38.09%) and C (54.05% and 41.66%, respectively). Plasma membrane integrity was similar after exposure to the three extenders (62.71% (SB), 68.76% (H), and 69.23% (C)). Mitochondrial activity was higher in SB immediately after dilution (SB= 1.05nm, H= 0.81nm, C= 0.79nm), and after 24 h (0.83nm (SB), 0.73nm (H) and 0.64nm (C)). After 48 h, the mitochondrial activity decreased to 0.72nm (SB), 0.69nm (H), and 0.63nm (C) (P > 0.05). The pH, osmolarity and pH after 48 h of cooling of the diluted semen were higher in SB (8; 382; 7.9), intermediate in H (7.5; 362; 7.32) and lower in C (7.16; 350; 7.07). Lipid peroxidation and its induction were similar in all groups. Data were analyzed by analysis of variance (ANOVA), and Duncan’s test was used to evaluate the significant differences (P < 0.05). Sodium bicarbonate reduced the progressive motility and increased the semen pH. The extender C was considered more appropriate for immediate use in artificial insemination. The non-buffered and HEPES-buffered extenders were considered appropriate for the cooling of equine semen for 48 h at 5°C.

Acupuncture Prior to and at Embryo Transfer in An Assisted Conception Unit – a Case Series

2006 ◽  
Vol 24 (1) ◽  
pp. 23-28 ◽  
Author(s):  
David Johnson
Keyword(s):  
Success Rate ◽  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Assisted Reproduction ◽  
Case Series ◽  
In Vitro Fertilisation ◽  
Assisted Conception ◽  
Before And After ◽  
In Pregnancy

Over a period of three years, acupuncture was offered to patients entering assisted reproduction therapy. Acupuncture sessions were given at varying, but usually weekly, intervals during the in vitro fertilisation (IVF) cycle, and immediately before and after embryo transfer. Twenty two patients (average age 36.2 years) were treated over a total of 26 IVF cycles and 15 pregnancies were achieved, as determined by presence of foetal heartbeat on ultrasound at four weeks post embryo transfer. This was a success rate of 57.7% compared with 45.3% for patients in the IVF unit not treated with acupuncture (P>0.05). Relaxing effects were noted following acupuncture and it is speculated that this may have contributed to the increase in pregnancy rate for the acupuncture group.

The relation to pregnancy, the child, motherhood of women in IVF situation

2015 ◽  
Vol 6 (4) ◽  
pp. 97-104
Author(s):  
Arina Yurevna Malenova ◽  
Irina Gennadevna Kytkova
Keyword(s):  
Family Relations ◽  
First Trimester ◽  
Family Education ◽  
In Vitro Fertilisation ◽  
Control Group ◽  
The Family ◽  
Before And After ◽  
Group 2 ◽  
High Level

Research objective - studying of features of the relation to pregnancy, the child, motherhood of women in IVF situation. Selection: 100 married pregnant women aged from 28 till 42 years (the first pregnancy of the first trimester, complications in the anamnesis isn't present) representing two groups on 50 people: 1) after artificial insemination (empirical group); 2) in a situation natural pregnancy (control group). The leading motives of pregnancy, types of the attitude towards themselves, pregnancies, to the child, people around, the prevailing installations in the sphere of the family relations, features of representation of future mothers about themselves and "the ideal parent" are defined by testing. Distinctions in all respects with women from control group are found. It is established that in vitro fertilisation the high level of readiness for motherhood according to its motivational characteristics is observed. Prevalence of constructive motives of pregnancy against concern in the health and aspirations to meet social expectations is revealed. The leading types of a gestational dominant are optimum and euphoric, the hypertrophied positive emotional background of pregnancy is observed. In the future of a bike probability the dependent relations with the child, preference of the sponsoring or authoritative style of family education. Revaluation of own parental qualities when comparing with image of ideal mother is observed. Results allow to carry women to the group of risk demanding psychological maintenance before and after the childbirth.

scholarly journals Maternal thyroid function in multifetal pregnancies before and after fetal reduction

2000 ◽  
Vol 164 (1) ◽  
pp. 7-11 ◽  
Author(s):  
O Ogueh ◽  
AP Hawkins ◽  
A Abbas ◽  
GD Carter ◽  
KH Nicolaides ◽  
...  
Keyword(s):  
Thyroid Function ◽  
Thyroid Stimulating Hormone ◽  
In Vitro Fertilisation ◽  
Fetal Reduction ◽  
Before And After ◽  
Group 2 ◽  
Maternal Thyroid ◽  
Twin Pregnancies ◽  
Selective Fetal Reduction

The aim of the study was to investigate maternal thyroid function in pregnancy by monitoring the circulating concentrations of thyroid stimulating hormone (TSH), free thyroxine (fT(4)) and human chorionic gonadotrophin (hCG) in multifetal pregnancies before and after embryo reduction. We studied two groups of women: group 1 comprised singleton (n=12) and twin (n=12) pregnancies achieved after superovulation and in vitro fertilisation and embryo transfer (IVF-ET), and group 2 were multifetal pregnancies (n=39) undergoing selective fetal reduction to twin pregnancies. Blood samples were obtained initially at 10-12 weeks gestation (before fetal reduction) and then 4 and 8 weeks afterwards. Before fetal reduction, the circulating concentrations of fT(4) in multifetal pregnancies were significantly greater than those in singleton or twin pregnancies (singleton, mean 16.49 pmol/l (interquartile range 14.09-18.13 pmol/l); twins, 15.84 (15.36-16.95 pmol/l); multifetal, 21.08 (16. 64-26.29 pmol/l); P<0.005 for singleton and twins), and in a multiple regression analysis, fT(4) was significantly related to the number of fetuses (F=23.739, P=0.0001), but not to hCG. After fetal reduction to twins, the circulating concentrations of fT(4) in multifetal pregnancies decreased progressively towards those in control twin pregnancies, but remained significantly greater at both 4 (P=0.003) and 8 weeks (P=0.050). This pattern of change in the concentrations of fT(4) is similar to, but lags behind, that of hCG, which attains twin levels 4 weeks after fetal reduction. This may represent a delayed thyroid response to the decreasing concentrations of hCG, but the alternative is that the maternal thyroid function is controlled by a fetal factor in addition to hCG.

scholarly journals 269IN VITRO FERTILITY OF BOAR SPERMATOZOA PRESERVED AT 10^°C FOR 22 DAYS

2004 ◽  
Vol 16 (2) ◽  
pp. 255
Author(s):  
H. Funahashi
Keyword(s):  
Oxidative Stress ◽  
Functional Status ◽  
Pregnancy Rate ◽  
Seminal Plasma ◽  
Artificial Insemination ◽  
Fluorescence Assay ◽  
Live Cells ◽  
Sperm Viability ◽  
Boar Spermatozoa

Fertility of boar spermatozoa as determined following artificial insemination seems to be maintained during liquid preservation at 10–15°C for several days, although prolonged liquid preservations reduce the pregnancy rate rapidly. However, it is not clear if spermatozoa can penetrate into oocytes in an IVF system even after a prolonged liquid preservation. Oxidative stress could also be one of the possible detrimental factors in liquid preservation of spermatozoa. In the present study, fertility of liquid-preserved spermatozoa was examined using an IVM-IVF system. Whether cysteine can improve the fertility was also determined. Spermatozoa (from four Berkshires) was resuspended at 1×108 cells mL−1 in Modena solution containing 15% (v/v) boar seminal plasma and 0 or 5mM cysteine after washing 3 times. Sperm suspensions (1mL) were then preserved at 10°C for 22 days following a program for cooling down (to 15°C for 4h, keeping at 15°C for 12h and then to 10°C for 6h). At Days 1, 8, 15 and 22 after the start of preservation, spermatozoa (5×105 cells mL−1) were co-cultured with IVM oocytes in an IVM/IVF system (Funahashi et al., 1997 Biol Reprod 57, 49–53). Viability and functional status of spermatozoa were also examined at Days 8 and 15 of preservation by using LIVE/DEAD sperm viability kit and CTC fluorescence assay. Data (mean±SEM) from 4–6 replicates were analyzed by ANOVA and Fisher’s protected LSD test. When spermatozoa that had been preserved without cysteine (Cys−) were used, penetration rates were not different (P&gt;0.05) from those with cysteine (Cys+) at Day 8 of preservation (91.4±3.4% in Cys− and 99.3±0.7% in Cys+), but lower (P&lt;0.02) at Days 15 and 22 (72.6±13.6% and 33.8±8.4% in Cys−; 94.8±2.1% and 71.1±10.8% in Cys+, respectively). Both viability and proportion of uncapacitated live cells were higher (P&lt;0.05) in Cys+ than Cys− at Days 8 and 15. These results demonstrate that boar spermatozoa can penetrate into oocytes in vitro even after a liquid preservation at 10°C for 22 days and that cysteine can improve the viability and penetrability in vitro of spermatozoa during liquid preservation. Supported by the Ito Foundation.

Leptin expression in adipose tissue from obese humans: depot-specific regulation by insulin and dexamethasone

1998 ◽  
Vol 275 (3) ◽  
pp. E507-E515 ◽  
Author(s):  
C. D. Russell ◽  
R. N. Petersen ◽  
S. P. Rao ◽  
M. R. Ricci ◽  
A. Prasad ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Subcutaneous Tissue ◽  
Obese Women ◽  
Omental Adipose Tissue ◽  
Before And After ◽  
Severely Obese ◽  
Specific Regulation ◽  
And Function ◽  
Leptin Expression

We investigated the in vitro regulation of leptin expression in adipose tissue from severely obese women and men before and after culture with insulin (7 nM) and/or dexamethasone (25 nM). Leptin mRNA and leptin secretion were two- to threefold higher in subcutaneous vs. omental adipose tissue before culture. Dexamethasone transiently increased leptin mRNA approximately twofold in both depots after 1 day of culture [ P < 0.01 vs. basal (no hormone control)], but leptin secretion was only increased in omental adipose tissue ( P < 0.005 vs. basal). Insulin did not increase leptin mRNA in either depot but increased leptin secretion ∼1.5- to 3-fold in subcutaneous tissue throughout 7 days of culture ( P < 0.05 vs. basal). The combination of insulin and dexamethasone increased leptin mRNA and leptin secretion approximately two- to threefold in both depots at day 1( P < 0.005 vs. basal or insulin) and maintained leptin expression throughout 7 days of culture. We conclude that insulin and glucocorticoid have depot-specific effects and function synergistically as long-term regulators of leptin expression in omental and subcutaneous adipose tissue from obese subjects.

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