Pregnancy and calving rates following transfer of in-vitro-produced river and F1 (river x swamp) buffalo (Bubalus bubalis) embryos in recipients on natural oestrus or synchronised for ovulation

The main objective of this study was to compare pregnancy and calving rates following transfer of in-vitro-produced fresh river and F1 (river x swamp) buffalo embryos in recipients synchronised by Ovsynch protocol or following natural oestrus. River embryos were produced from cumulus-oocyte complexes (COCs) derived by ovum pick up (OPU) on 40 Murrah and Nili-Ravi donor buffaloes over a twice-weekly collection schedule for 120 single OPUs. F1 embryos were produced by fertilisation of swamp COCs recovered from abattoir ovaries coincubated with river sperm cells. Both groups of embryos were produced following the same protocol for in vitro production. With regard to the OPU source of COCs, 923 antral follicles were punctured and 647 COCs were recovered (70%). From 462 selected COCs for IVM, 257 (55.6%) cleaved zygotes were recorded leading to 93 blastocysts (20.1%). In total, 590 swamp COCs were aspirated from abattoir ovaries and 476 were selected for IVM leading to 270 (56.7%) cleaved zygotes and resulting in 137 blastocysts (28.8%). River and F1 embryos were transferred between Day 6 to 7 of in vitro development, corresponding to blastocyst-expanding blastocyst, into F1 recipients synchronised by Ovsynch and swamp buffaloes following natural oestrus, respectively, each of them receiving two embryos. According to palpation per rectum of the ovaries at the time of embryo transfer, 26 of the 47 (55.3%) F1 recipients synchronised by Ovsynch were considered suitable for transfer, resulting in seven pregnancies (26.9%) and four calvings (15.3%) owing to three abortions occurring between 2 and 3 months of pregnancy. In total, 29 swamp recipients following natural oestrus were judged suitable as recipients, resulting in 12 pregnancies (41.4%) and 10 calvings (34.5%) owing to two abortions at 2 and 3 months of gestation respectively. Pregnancy and calving rates following transfer of river and F1 embryos were similar. Likewise, weight at birth of calves derived from transfer of river and F1 embryos was not different: 30.5 +/- 1.4 and 32.9 +/- 2.4 respectively. Pregnancy and calving rates following AI in a group of river and swamp buffaloes considered for reference in this study were similar to recipients carrying in-vitro-produced embryos. Collectively, no apparent postnatal complications were recorded in resulting live calves.     

Role of preovulatory concentrations of estradiol on timing of conception and regulation of the uterine environment in beef cattle

ABSTRACT

The ability to induce ovulation with an injection of GnRH facilitated the development of fixed-time artificial insemination (AI) protocols. However, animals that exhibit estrus prior to fixed-time AI have greater pregnancy success. Thus, the objectives of the present experiments were to determine the impact of estrus expression prior to fixed-time AI on timing of conception and to characterize the role of preovulatory estradiol in regulating changes in expression of uterine genes. In experiment 1, data were collected from 4,499 beef cows inseminated by fixed-time AI in 31 different herds. Animals that did not conceive to AI but exhibited estrus before timed-AI were more likely to conceive during cycle 1 after AI, and overall conceived earlier in the breeding season compared to animals that did not exhibit estrus. In experiment 2, beef cows were synchronized using a fixed-time AI protocol. Uterine horn biopsies and blood samples were collected on Day 0, 5, 10, or 16. Concentrations of estradiol on Day 0 did not influence expression of progesterone receptor, ER beta, or oxytocin receptor. Increased concentrations of estradiol on Day 0 increased expression of ER alpha from Days 0 to 5 of the estrous cycle. Furthermore, cows with increased concentrations of estradiol on Day 0 had increased expression of inhibin beta A, and uterine milk protein precursor. Thus, animals that do not exhibit estrus prior to fixed-time AI had decreased breeding season pregnancy success and conceived later in the breeding season, and preovulatory concentrations of estradiol likely play a major role in this establishment of pregnancy, not only directly by regulating uterine gene expression, but also indirectly throughout the subsequent estrous cycle.     

Evaluation of follicular oxidant-antioxidant balance and oxidative damage during reproductive acyclicity in water buffalo (Bubalus bubalis

ObjectiveTo investigate changes in follicular fluid concentrations of reactive oxygen species (ROS) and total antioxidant capacity (TAC) and degree of oxidative damage to follicular cells, using protein carbonyl (PC) as marker of oxidative stress, were investigated during reproductive acyclicity in buffalo.MethodsFollicular fluid was aspirated from follicles grouped into three classes depending upon their diameter [small (5.0–7.0 mm), medium (7.1–10.0 mm), and large (>10.0 mm)]. Progesterone and estradiol were estimated to determine functional status (P:E ratio) of the follicles.ResultsAcyclic buffaloes had greater concentrations of ROS (P<0.001) and PC (P=0.0412) and lower concentrations of TAC (P=0.0280) than cyclic buffaloes. An interesting novel finding was the complete absence of low P:E functionally active follicles in acyclic buffaloes. Results indicated a pronounced follicular fluid oxidant-antioxidant imbalance and oxidative damage to follicular cells during acyclicity in buffalo.ConclusionIn conclusion, this study provided evidence about role of oxidative stress in pathogenesis of reproductive acyclicity.     

Getting to Know Our Guests: Characterizing the Reproductive Microbiome by Next Generation Sequencing

Abstract

The objective of this study was to determine whether there are changes in primary, primordial, and growing follicles after dehydroepiandrosterone (DHEA) administration in rats that have diminished ovarian reserve (DOR) due to 4-vinylcyclohexene diepoxide (VCD) application, and to examine the mechanism of the probable effect of DHEA on folliculogenesis. Two groups of Wistar rats were used. In Group A unilateral oophorectomy (eight rats) was carried out on day-0. The remaining study ovary was removed by relaparotomy after VCD (160?mg/kg, intraperitoneally) was administered for 15 days. In Group B unilateral oophorectomy (eight rats) was carried out on day-0. The remaining study ovary was removed by relaparotomy after VCD (160?mg/kg, intraperitoneally) administration for 15 days followed by DHEA (60?mg/kg body weight) daily for 45 days. Primordial, primary, and growing (secondary?+?antral) follicles were counted in 1,664 sections from 32 ovaries. In all three types of follicles (primordial, primary, and growing), the number of follicles significantly decreased in the study ovaries compared to the control ovaries in both Group A and Group B. In Group B, atresia rates were significantly lower in the study ovary compared to the control ovary in all of the follicular groups: primordial (p?=?0.02), primary (p?=?0.01), and growing (p?=?.027). To demonstrate the probable effects of DHEA on follicular dynamics, we also compared the study ovaries in both groups; the primordial (p?=?0.027), primary (p?=?0.031), and growing (p?=?0.04) number of follicles were significantly higher in Group B compared to Group A. In conclusion, our findings suggest that DHEA administration in DOR rats due to VCD results in a larger follicular pool. Decreased atresia may be one of the possible effects of DHEA in DOR cases. Whatever the mechanism, DHEA treatment potentially may be useful clinically as a means to increase the number of gonadotropin-responsive follicles for ovarian stimulation.     

The fate of frozen human embryos when transferred either on the day of thawing or after overnight culture

ObjectiveTo study the performance of thawed zygotes and cleavage stage embryos transferred either on the day of thaw or after overnight culture.MethodsA retrospective study of 864 frozen embryo transfer cycles. Cryosurvival rates per thawed embryo and implantation rates were analysed for embryos frozen on Day 1, Day 2 or Day 3 relative to oocyte collection (Day 0) and transferred on the day of thaw or after overnight culture, together with clinical pregnancy rates and prevalence of multiple gestations.ResultsSurvival of Day 3 embryos was significantly lower than those frozen on Day 1 (P=0.017) or Day 2 (P=0.015). Following overnight culture, resumption of mitosis of zygotes was more frequent than Day 2 (P=0.000) which are in turn higher than Day 3 (P=0.000) embryos. The implantation rate for Day 2 embryos dividing overnight was significantly higher than those that did not divide for women <35 yrs (P=0.001) but not those women <35 yrs (P=0.055). There were no differences in the implantation rates for those dividing or not after culture, for embryos frozen on Day 3 for women <35 yrs (P=0.254) or <35 yrs (P=0.403).ConclusionsLater cleavage stage post-thaw embryos survive and resume mitosis less frequently compared to earlier stages. Embryos not resuming mitosis after culture overnight can implant, particularly Day 3 embryos, suggesting that they can further increase the cumulative pregnancy rate per oocyte collection and that discarding them is wasteful. Overnight culture is best used for logistical reasons rather than a strategy to improve pregnancy rates.     

Investigation on some biochemical parameters and effect of hormonal treatment in anoestrous dairy cows with cystic ovarian follicle

ObjectiveTo investigate some blood biochemical parameters in anoestrous dairy cows with cystic ovarian follicles (COF) during the first two months post partum and the effect of hCG-PGF2_α -GnRH treatment on their reproductive response.MethodsThe investigation was performed on 20 lactating Holstein cows, divided into two groups: control group (n=8) and treated group (n=12). All animals had cystic ovarian follicles, detected during the two consecutive transrectal ultrasound examinations at 7-day interval. Blood samples from each animal were collected on the day of second ultrasonography (Day 0) and subjected to a hormonal and biochemical analysis. Treated cows were subjected to another ultrasound examination on day of prostaglandin injection and the presence of corpus luteum was recorded. The control animals were not treated, while the treated group received 1 500 IU hCG on day 0, PGF2_α 500 mg on day 7 and 100 μg GnRH 48 hours latter. Eighteen hours after GnRH administration animals bearing corpus luteum on Day 7 were artificiallly inseminated. The last ultrasound examination was made forty days after start of the threapy. On the base of the obtained results, cumulative oestrous activity, cumulative ovulation rate, pregnancy rate and COF persistence were determined.ResultsThe blood analysis in the both groups on Day 0 showed significantly (P<0.05) lower mean concentration of progesterone, glucose and inorganic phosphate compared to the reference range values for cyclic dairy cows. Cumulative oestrous activity (91.7%) and cumulative ovulation rate (75.0%) were considerably higher (P<0.05) in treated group than control group, whereas the cystic follicle persistence (25.0%) was lower (P<0.05) in the treated group than control group.ConclusionsThe present study confirmed the thesis for low blood levels of glucose and inorganic phosphate in cows presenting cystic ovarian follicles. It also indicated significant improvement of cumulative oestrous activity and ovulation rate in animals subjected to hCG-PGF2_α -GnRH administration than non-treated animals. The used hormonal protocol could decrease cystic ovarian follicle persistence in dairy cows.     

Relationship between processed total motile sperm count of husband or donor semen and pregnancy outcome following intrauterine insemination

We retrospectively analyzed 6,360 artificial insemination cycles of husband's semen through intrauterine insemination (AIH-IUI) or artificial insemination with donor semen through intrauterine insemination (AID-IUI) in patients with infertility between August, 1998 and August, 2010. The relationship between processed total motile sperm count (PTMS) and pregnancy outcome was determined. The study was divided into 6 groups according to PTMS. Group 1:?≤?2.0 million, Group 2: 2.1-4.0 million, Group 3: 4.1-6.0 million, Group 4: 6.1-8.0 million, Group 5: 8.1-10.0 million, and Group 6: >10.0 million. There was no statistically significant difference in age, duration of infertility, unilateral tubal patency, induced ovulation, and single IUI or double IUI between the 6 groups in both AIH-IUI and AID-IUI. The total clinical pregnancy rate of AIH-IUI was 10.81 % and AID-IUI was 27.52 %. Among the 6 groups, the clinical pregnancy rate was the lowest in Group 1 (P?<?0.05) in both AIH-IUI and AID-IUI. With the increased PTMS, the clinical pregnancy rate of IUI was improved. However, a statistical difference between groups was only observed for Group 1. When PTMS is?≤?2?×?10⁶ the clinical pregnancy rate of IUI is significantly decreased. In this case in vitro fertilization (IVF) should be adopted.     

In vitro and in vivo regulation of follicular formation and activation in cattle

The establishment of a stockpile of non-growing, primordial follicles and its gradual depletion through activation of primordial follicles are essential processes for female fertility. However, the mechanisms that regulate follicle formation, the activation of primordial follicles to begin growth and the primary-to-secondary follicle transition are poorly understood, especially in domestic animals and primates. The authors' laboratory is engaged in studying early stages of follicular development in cattle and this review summarises the progress to date. Bovine follicles begin to form in fetal ovaries around the beginning of the second trimester of pregnancy (about Day 90), but the first activated, primary follicles do not appear until after Day 140. Bovine fetal ovaries produce steroids and production is highest during the first trimester. In vitro, oestradiol and progesterone inhibit follicle formation and acquisition by newly formed follicles of the capacity to activate. Meiotic arrest of the oocyte in the diplotene stage of first prophase does not occur until after follicle formation and is correlated with acquisition of the capacity to activate. This may explain the gap between follicle formation and appearance of the first activated follicles. Once capacity to activate has been acquired, it seems likely that activation in vivo is controlled by the balance between stimulators and inhibitors of activation. Insulin and kit ligand stimulate and anti-Müllerian hormone (AMH) inhibits activation in vitro. Few bovine follicles transition from the primary to the secondary stage in vitro, but this transition is increased by medium supplements, testosterone and vascular endothelial growth factor (VEGF).     

Ovarian size and vascularization as assessed by three-dimensional grayscale and power Doppler ultrasound in asymptomatic women 20-39 years old using combined oral contraceptives

BackgroundThe aim of this study is to estimate ovarian volume, number and volume of antral follicles, and ovarian power Doppler vascular indices as assessed by three-dimensional (3D) transvaginal grayscale and power Doppler ultrasound in women using combined oral contraceptives (COC).Study DesignTwo hundred thirteen gynecologically asymptomatic women 20–39 years old using COC were examined with transvaginal 3D grayscale and power Doppler ultrasound on cycle day 4–8 (first cycle day is first day of withdrawal bleeding). We used a Voluson E8 ultrasound system with a 6–12-MHz transvaginal transducer. Ovarian volume, number and volume of antral follicles ≥2 mm, vascularization index (VI), flow index (FI) and vascularization flow index (VFI) were calculated using the virtual organ computer-aided analysis (VOCAL?) and sonography-based automated volume calculation (SonoAVC?) software. Results are described separately for women with follicles 2.0–10.0 mm and for those with at least one follicle >10.0 mm for two age groups: 20–29 years (n=166) and 30–39 years (n=47). Results are also compared between women on monophasic (n=151) and triphasic (n=59) COC, and between women using COC with older (n=110) and newer (n=100) progestins and different doses of estrogen.ResultsOne hundred eighty-nine (89%) women had follicles 2.0–10.0 mm, and 24 (11%) had follicle(s) >10.0 mm. The proportion of women with follicle(s) >10.0 mm did not differ between women with different types of COC. In women with follicles 2.0–10.0 mm, the right ovary was larger (mean difference 0.5 cm³ [95% confidence interval 0.22–0.82]) and contained more follicles (mean difference 1.5 [0.52–2.56]) than the left one in the age group 20–29 years. The same differences between the right and left ovary were seen in women 30–39 years old, but they were not statistically significant. In both ovaries, the number of antral follicles 2.0–10.0 mm [median (range)] was significantly higher in women 20–29 than in those 30–39 years old [11 (2–34) vs. 8 (1–26), p=.012 for the right ovary; 9 (0–28) vs. 7 (1–28), p=.035 for the left ovary]. Ovarian volume tended to be smaller in women 20–29 than in those 30–39 years old, but the differences were not statistically significant. Size of the largest follicle, total follicular volume and vascular indices manifested no clear differences between the age groups. For all 378 ovaries with follicles ≤10 mm, ovarian volume ranged from 1 to 16 cm³ (median 5), total follicular volume ranged from 0.03 to 2.7 cm³ (median 0.7), VI ranged from 0.0 % to 13.4% (median 0.97), FI ranged from 0 to 38 (median 25), and VFI ranged from 0.0 to 4.7 (median 0.3).ConclusionsOur results show estimated ranges of 3D grayscale and power Doppler ultrasound measurements in ovaries of women using COC.     

Chronic exposure to fine particulate matter emitted by traffic affects reproductive and fetal outcomes in mice

Air pollution is an important environmental health risk factor that can result in many different gestational and reproductive negative outcomes. In this study, we have investigated the effects of two different times of exposure (before conception and during pregnancy) to urban ambient particulate matter on reproductive and pregnancy outcomes in mice. Using exposure chambers receiving filtered (F) and non-filtered (NF) air, we observed that exposed females exhibited changes in the length of estrus cycle and extended estrus and, therefore, a reduction in the number of cycles during the studied period (F 2.6±0.22 and NF 1.2±0.29, p=0.03). The mean number of antral follicles declined by 36% (p=0.04) in NF mice (75±35.2) compared to F mice (118.6±18.4). Our results further indicate a significant increase in time necessary for mating and decreased fertility and pregnancy indices (p=0.003) in NF couples. Mean post-implantation loss rates were increased by 70% (p?0.005) in the NF2 group (exposed before and during pregnancy to NF air) compared to the F1 group (exposed before and during pregnancy to F air) and were influenced by both pre-gestational (p<0.004) and gestational (p<0.01) period exposure. Fetal weight was significantly higher in the F1 group when compared with the other groups (p<0.001), at a 20% higher weight in the F1 group (0.86±0.18 g) than in the NF2 group (0.68±0.10 g). Furthermore, fetal weight was influenced by both pre-gestational and gestational period exposure, and a significant interaction between these two factors was found (p<0.001). This study demonstrated that exposure to ambient levels of urban traffic-generated particulate matter negatively affects different functions and stages of the reproductive process. Our results also reinforce the idea that maternal exposure to air pollution is linked to negative pregnancy outcomes, even if the exposure occurs only before conception.     

A randomized phase 3 trial of the immunogenicity and safety of coadministration of a live-attenuated tetravalent dengue vaccine (TAK-003) and an inactivated hepatitis a (HAV) virus vaccine in a dengue non-endemic country

BackgroundVaccination against hepatitis A virus (HAV) is largely recommended for travelers worldwide. Concurrent dengue and HAV vaccination may be desired in parallel for travelers to countries where both diseases are endemic. This randomized, observer-blind, phase 3 trial evaluated coadministration of HAV vaccine with tetravalent dengue vaccine (TAK-003) in healthy adults aged 18–60 years living in the UK.MethodsParticipants were randomized (1:1:1) to receive HAV vaccine and placebo on Day 1, and placebo on Day 90 (Group 1), TAK-003 and placebo on Day 1, and TAK-003 on Day 90 (Group 2), or TAK-003 and HAV vaccine on Day 1, and TAK-003 on Day 90 (Group 3). The primary objective was non-inferiority of HAV seroprotection rate (anti-HAV ≥ 12.5 mIU/mL) in Group 3 versus Group 1, one month post-first vaccination (Day 30) in HAV-naïve and dengue-naïve participants. Sensitivity analyses were performed on combinations of baseline HAV and dengue serostatus. Secondary objectives included dengue seropositivity one month post-second vaccination (Day 120), HAV geometric mean concentrations (GMCs), and safety.Results900 participants were randomized. On Day 30, HAV seroprotection rates were non-inferior following coadministration of HAV and TAK-003 (Group 3: 98.7 %) to HAV administration alone (Group 1: 97.1 %; difference: ?1.68, 95 % CI: ?8.91 to 4.28). Sensitivity analyses including participants who were neither HAV-naïve nor DENV-naïve at baseline supported this finding. Anti-HAV GMCs on Day 30 were 82.1 (95 % CI: 62.9–107.1) mIU/mL in Group 1 and 93.0 (76.1–113.6) mIU/mL in Group 3. By Day 120, 90.9–96.8 % of TAK-003 recipients were seropositive (neutralizing antibody titer > 10) to all four dengue serotypes. Coadministration of HAV vaccine and TAK-003 was well tolerated, with no important safety risks identified.ConclusionImmune responses following coadministration of HAV vaccine and TAK-003 were non-inferior to administration of HAV vaccine alone. The results support the coadministration of HAV vaccine and TAK-003 with no adverse impact on immunogenicity, safety, and reactogenicity of either vaccine.ClinicalTrials.gov registration: NCT03525119.     

Effect of increasing progesterone concentration from Day 3 of pregnancy on subsequent embryo survival and development in beef heifers

Higher systemic progesterone in the immediate post-conception period is associated with an increase in embryonic growth rate, interferon-tau production and pregnancy rate in cattle. The objective of this study was to examine the effect of increasing progesterone concentration on Day 3 on subsequent embryo survival and development. Oestrus (Day 0) was synchronised in beef-cross heifers (n=210) and approximately two-thirds of the heifers were inseminated with semen from a proven sire, while the remainder were not inseminated. In order to produce animals with divergent progesterone concentrations, half of the animals received a progesterone-releasing intravaginal device (PRID) on Day 3 of the oestrous cycle, which was left in situ until slaughter. The four treatment groups were: (i) pregnant, high progesterone; (ii) pregnant, normal progesterone; (iii) non-pregnant, high progesterone; and (iv) non-pregnant, normal progesterone. Animals were blood-sampled twice daily from Days 0 to 8 and once daily thereafter until slaughter on Days 5, 7, 13 or 16, corresponding to the 16-cell stage, the blastocyst stage, the beginning of elongation and the day of maternal recognition of pregnancy, respectively. Embryos were recovered by flushing the tract with phosphate-buffered saline and characterised by stage of development and, in the case of Days 13 and 16, measured. Data were analysed by mixed models ANOVA, Chi-square analysis and Student's t-test where appropriate. Insertion of a PRID on Day 3 increased (P<0.05) progesterone concentrations from Day 3.5 onwards. There was no difference between treatments in the proportion of embryos at the expected stage of development on Days 5 or 7 (P>0.05). While not significantly different, the proportion of viable embryos recovered was numerically greater in the high progesterone group on both Day 13 (58 v. 43%) and Day 16 (90 v. 50%). Elevation of progesterone significantly increased embryonic length on Day 13 (2.24+/-0.51 mm v. 1.15+/-0.16 mm, P=0.034) and Day 16 (14.06+/-1.18 cm v. 5.97+/-1.18 cm, P=0.012). In conclusion, insertion of a PRID on Day 3 of the oestrous cycle increased serum progesterone concentrations on subsequent days, which, while having no phenotypic effect on embryonic development on Days 5 or 7, was associated with an increase in embryonic size on Days 13 and 16.     

Effect of coincubation time of sperm-oocytes on fertilization,embryonic development,and subsequent pregnancy outcome

Several studies have reported improved IVF by shortening the time of sperm-oocyte coincubation from 16–18 hours to 1–4 hours. The objective of this study was to examine the advantages and disadvantages of a shortened sperm-oocyte coincubation time in order to assess the effects of this insemination method for clinical IVF practice. Two insemination methods, the shortened method (4 hours) and the standard method (16–18 hours) of coincubation of sperm-oocytes for two groups of patients based on the quality of sperm were compared. Group I, was composed of couples without male factor; Group II, involved couples with mild male factor. Fertilization, good quality embryos, clinical pregnancy, and implantation rates were compared by two different insemination methods. In Group I, fertilization, clinical pregnancy, and implantation rates were not different between the two insemination methods. However, the polyspermy rate was significantly higher (P?<?0.05) in the shortened (7.3%) than in the standard (4.1%) insemination method. In Group II, the fertilization rate was significantly lower (P?<?0.05) using the shortened insemination method (62.6%) compared to the standard insemination method (68.7%). When fertilization failed with the shortened insemination method, the clinical pregnancy and implantation rates were 34.7% and 24.1%, respectively, from the rescue intracytoplasmic sperm injection (ICSI). The live birth rate from the rescue ICSI was 32.0% with normal infants. The duration of sperm-oocyte coincubation does not affect fertilization, embryo quality, clinical pregnancy, and implantation rates. However, fertilization rates will decrease with the shortened insemination method when the sperm parameters are poor. From the results of the present study we suggest that the combination of the shortened sperm-oocyte coincubation and rescue ICSI method may be an efficient method for IVF treatment in order to prevent fertilization failure when sperm parameters were poor as mild male factor.     

Relationship between processed total motile sperm count of husband or donor semen and pregnancy outcome following intrauterine insemination

We retrospectively analyzed 6,360 artificial insemination cycles of husband's semen through intrauterine insemination (AIH-IUI) or artificial insemination with donor semen through intrauterine insemination (AID-IUI) in patients with infertility between August, 1998 and August, 2010. The relationship between processed total motile sperm count (PTMS) and pregnancy outcome was determined. The study was divided into 6 groups according to PTMS. Group 1:?≤?2.0 million, Group 2: 2.1-4.0 million, Group 3: 4.1-6.0 million, Group 4: 6.1-8.0 million, Group 5: 8.1-10.0 million, and Group 6: >10.0 million. There was no statistically significant difference in age, duration of infertility, unilateral tubal patency, induced ovulation, and single IUI or double IUI between the 6 groups in both AIH-IUI and AID-IUI. The total clinical pregnancy rate of AIH-IUI was 10.81 % and AID-IUI was 27.52 %. Among the 6 groups, the clinical pregnancy rate was the lowest in Group 1 (P?相似文献   

Reproductive characteristics and corticoid levels of female white-footed mice fed ad libitum and restricted diets containing a polychlorinated biphenyl.

Sixty female white-footed mice (Peromyscus leucopus) were fed either ad libitum or 70% ad libitum diets for 5 weeks. During the last 3 weeks of feeding a polychlorinated biphenyl (Aroclor 1254) was incorporated into the diets at levels of 0,25, or 100 ppm (based on ad libitum intake). Polychlorinated biphenyl ingestion increased liver weights, reduced pentobarbital-induced sleep times, and generally reduced plasma corticoid levels but had no effect on reproductive organ weights or the incidence of estrus (as judged by vaginal smears). Feed restriction caused marked atrophy of the ovaries and uterus and reduced the incidence of estrus.     

The viability of transferred sheep embryos after long-term cryopreservation

The survival to term of 414 sheep embryos, thawed and transferred after conventional cryopreservation and storage for 13 years, was evaluated. A concurrent experiment involving the transfer of vitrified embryos to 91 ewes and artificial insemination of 51 ewes with frozen-thawed semen from sires of the long-term cryopreserved embryos provided forms of control treatments. The donor ewes had a mean ovulation rate of 10.9, and 7.1 embryos per ewe were cryopreserved. Each recipient ewe received two embryos and pregnancy was assessed at Day 18, Day 54 and term. The pregnancy rate was lower in the long-term embryo group than the artificial insemination group at Day 18 (P<0.01) and Day 54 (P<0.05), although the difference at term (31% v. 49%) was not significant, with the vitrified embryo group being similar to the long-term group. Embryo survival to birth was 21%, with the majority of loss (80%) occurring by Day 18. The later stage of development and higher grade of transferred embryos and the older age of donor ewes resulted in a significantly higher (P<0.01) pregnancy rate at Day 54 and term, and percentage of lambs born and weaned. Other effects of donor ewes (genotype, superovulation treatment, number of ovulations and embryos cryopreserved) were not significant. Implications for the design of genetic evaluation and germplasm conservation programmes using embryo cryopreservation technology are discussed.     

Improvement of embryonic development and clinical outcomes of germinal vesicle stage oocytes using a microvibration culture system

ABSTRACT

In vitro maturation (IVM) has evolved as a clinical treatment option in assisted reproductive technology. However, the poor developmental potential of germinal vesicle (GV)-stage oocytes is still suboptimal. This study’s objective was to evaluate the effect of a microvibration culture system (MVC) during IVM and/or in vitro culture (IVC) on the clinical outcomes and the embryonic development potential of human GV-stage oocytes collected from human chorionic gonadotropin (HCG)-primed IVM and fertilization-embryo transfer (IVM/F-ET) cycles of patients with polycystic ovaries (PCO). A total of 206 HCG-primed IVM/F-ET cycles were divided into four groups according to the microvibration and static culture system applied during IVM and/or IVC: Group SS (static system during both IVM and IVC); Group SV (static system during IVM alternated with microvibration system during IVC); Group VS (microvibration system during IVM alternated with static system during IVC), and Group VV (microvibration system during both IVM and IVC). The results indicate that the rates of in vitro MII oocytes per cycle, fertilization, and cleavage were not significantly different between the groups. The rate of good-quality embryos in Group SV tended to be higher than the rate in Groups SS and VS, but there was no significant difference between Group SS and Group SV. Clinical pregnancy, implantation, and live birth rates of Groups SV and VS were slightly higher than those of Group SS. However, the rate of good-quality embryos with at least six cells on day 4, the clinical pregnancy, implantation, and live births in Group VV were significantly higher than those in Group SS. These results indicate that, compared with the static culture system, the MVC system applied for both IVM and IVC seems to improve the clinical outcomes and the quality of embryos of GV oocytes derived from HCG-primed IVM/F-ET cycles in PCO patients.

Abbreviations: PCO: polycystic ovaries; HCG: human chorionic gonadotropin; GV: germinal vesicle; MII: metaphase II; IVM: in vitro maturation; IVF: in vitro fertilization; IVC: in vitro culture: MVC: microvibration culture; SC: static culture; ICSI: intracytoplasmic sperm injection; IVM/F-ET: IVM and fertilization-embryo transfer; AMH: anti-Mullerian hormone; OHSS: ovarian hyperstimulation syndrome     

Histological Changes of the Ovary in Pregnant Mice Vaginally Exposed to Toxoplasma gondii

Background:

Congenital toxoplasmosis is one cause of abortion. Infection can disrupt ovarian cycles and because toxoplasmosis is an infectious disease may have a similar effect on the ovaries. The purpose of this study was to investigate the pathological changes in the ovaries due to toxoplasmosis.

Methods:

Tachyzoites of Toxoplasma gondii were harvested from peritoneal fluid of mice, experimentally infected. Two females and one male mouse were housed per cage for mating in the overnight. The pregnant mice were divided into experimental and control groups. Experimental group were infected by parasite but the control group received the normal saline. The experimental and control mice were euthanized. Ovaries and uterine horns of animals were removed and prepared for light microscopy.

Results:

Ovaries of infected pregnant mice presented gross morphological differences compared to the control groups. In ovaries of experimental groups, changes of corpus luteum were observed. The comparison of experimental and control groups revealed that the number of primary follicles, secondary follicle, atretic primary follicles and atretic secondary follicles had significant differences (P≤0.001).

Conclusion:

Toxoplasma gondii alters ovarian follicular growth and development in mice. In addition, it alters number of different phases of follicles and corpus luteum in ovaries of mice.     

Basal serum anti-müllerian hormone and antral follicle count are predictors of ovarian response for Asian women in Singapore

ObjectiveTo determine the basal ovarian reserve markers for prediction of poor and high responses and clinical pregnancy outcome in subfertile Asian women requiring controlled ovarian stimulation (COS) treatment in Singapore.MethodsSubfertile Asian women, aged ≤40 with basal serum follicle stimulating hormone (FSH) level of ≤12 IU/L, were enrolled prospectively during routine preliminary endocrine ovarian reserve assessment prior to COS regime for intracytoplasmic sperm insemination (ICSI) cycles. Basal serum levels of the endocrine ovarian reserve markers (anti-Müllerian hormone (AMH), estradiol, FSH, luteinizing hormone), antral follicle count (AFC) and ovarian response parameters were compared between the Poor (retrieved oocytes ≤4), Normal (retrieved oocytes, 5 to 19) and High Responder (retrieved oocytes ≥20) groups of women.ResultsBasal serum AMH and AFC were significantly correlated to age (r =−0.213 and −0.243, respectively) and to the number of retrieved oocytes (r=0.570 and 0.523, respectively) (P<0.05). Both basal serum AMH and AFC were significant discriminators of poor response (cut-off levels of ≤2.0 µg/L and ≤12, respectively) and for high response (AMH ≥3.2 µg/L and AFC of ≥20, respectively) to COS. Basal AMH was the only significant predictor for clinical pregnancy outcome, ROCAUC =0.71, cut-off level of ≥3.0 µg/L and odds ratio of 1.42.ConclusionBoth basal serum AMH and AFC were reliable ovarian reserve markers for predicting poor and high ovarian response to COS in Asian women. Basal AMH was the only significant predictor for clinical pregnancy outcome.     

Vaginal contraceptive activity of a chelated vanadocene

Bis(cyclopentadienyl) complexes of vanadium (IV) or vanadocenes are rapid and potent inhibitors of human sperm motility with potential as a new class of contraceptive agents. This study sought to determine the vaginal contraceptive activity of vanadocene dithiocarbamate (VDDTC), a stable vanadocene (IV)-chelated complex, using the standard rabbit model as well as the domestic pig as a physiologically relevant animal model for contraception. In experiment I, ovulating New Zealand White does in subgroups of eight were artificially inseminated (AI) with semen mixed with VDDTC (0.01-1 mM) or vehicle. In experiment II, ovulating does in subgroups of 18 were AI at 5 and 60 min after intravaginal application of a gel with and without 0.1% VDDTC or 2% nonoxynol-9 (N-9) (Gynol II, Ortho Pharmaceutical, Raritan, NJ), and allowed to complete term pregnancy. In experiment III, seven sexually mature Duroc gilts in standing estrus were AI with and without intravaginal application of 0.1% VDDTC gel microemulsion. Exposure of rabbit semen to VDDTC at the time of artificial insemination resulted in a dose-dependent reduction in fertility. Exposure of semen to 1 mM VDDTC led to complete inhibition of fertility as assessed by the number of embryos (control 49/94 vs. VDDTC-treated 0/117, p<.0001) or the percent embryos (52% vs. 0%, respectively) based on number of embryos to corpora lutea. Intravaginal administration of 0.1% VDDTC gel microemulsion or Gynol II prior to artificial insemination significantly inhibited term pregnancy rates (88% and 62% inhibition, respectively) when compared to control gel alone. Vanadocene dithiocarbamate gel microemulsion provided 80% inhibition of fertility even when insemination was delayed until 60 min after intravaginal application of VDDTC gel microemulsion. Rabbits that delivered litters despite intravaginal exposure of semen to VDDTC via gel microemulsion had healthy offsprings with no apparent perinatal repercussions. In domestic pigs, intravaginal applications of 0.1% VDDTC gel microemulsion prior to artificial insemination led to complete inhibition of fertility as assessed by the number of embryos (control 29/52 vs. VDDTC-treated 0/44, p<.0001) or the percent embryos (56% vs. 0%, respectively) based on the number of embryos to corpora lutea. These results suggest that VDDTC is a potent contraceptive agent in vivo. Intravaginal use of VDDTC via a gel microemulsion has clinical potential as a safe alternative to currently used detergent-type contraceptives.