Total Number Of Embryos Research Articles

Serum progesterone on the day of human chorionic gonadotropin (hCG) trigger as a predictor of in-vitro fertilization (IVF) outcome - a retrospective analysis of seven years.

To investigate the relationship between progesterone (P4) levels on the day of hCG trigger and IVF outcomes. This is a retrospective analysis of IVF cycles from January-2013 to December-2019 from a single center. Women (21-39 years) submitted to IVF treatment for various infertility factors were included, while donor oocyte cycles and cancelled cycles were excluded from the study. The primary outcome measure was live birth rate. A total of 2149 cycles were analyzed. Of these, 223 (10.38%) were in the low P4 group (<0.5 ng/ml), 1163 (54.12%) in the normal P4 group (0.5-1.5 ng/ml), and 763 (35.50%) in the high P4 group (>1.5ng/ml). The groups were comparable with respect to age, factor of infertility and baseline AMH. The antagonist protocol was significantly more prescribed to the high P4 group (p<0.001). Live birth rates were 14.4%, 21.6%, and 21% (p<0.001), respectively, in three groups. Univariate analysis found that total cetrotide dose, total number of retrieved and fertilized oocytes, total number of embryos formed, transferred, and vitrified, and P4 on the day of hCG (p<0.001) were statistically significant after adjusting for age and BMI. In multivariate logistic regression after adjusting for age and BMI, only high P4 (aOR:0.60; p<0.001), total cetrotide dose (aOR: 0.82; p<0.001), and total utilizable embryos (aOR:1.11; p=0.029) were statistically significant. Having an elevated serum progesterone level on the day of hCG trigger was associated with lower pregnancy rates, but this is still not a robust marker to predict live births. More good quality evidence is needed.

Genetic relationship between superovulatory response traits and carcass traits in Japanese Black cattle.

We estimated the genetic correlations between superovulatory response traits and carcass traits in Japanese Black cattle. As regards the superovulatory response traits in cows, we analyzed the phenotypic records of the total number of embryos and oocytes (TNE) and the number of good embryos (NGE) collected from 1532 donors between 2008 and 2018. As regards the carcass traits in fattened animals, we analyzed the phenotypic records for cold carcass weight, rib eye area, rib thickness, subcutaneous fat thickness, estimated yield percent, and marbling score for 1448 progenies derived from 596 donors and slaughtered between 2004 and 2020. Variance components were estimated using single-trait and two-trait animal models and the restricted maximum likelihood approach. The estimated genetic correlations with the carcass traits ranged from -0.05 to 0.04 for TNE and from -0.14 to 0.04 for NGE, and their standard errors ranged from 0.10 to 0.14. These results imply that the genetic relationship between the superovulatory response traits in Japanese Black donor cows and the carcass traits in their fattened progenies was weak to negligible. Therefore, we concluded that selecting donors with superior genetic ability for superovulatory responses would not have antagonistic effects on carcass performance in their fattened progenies.

​Effect of Season on Quantity and Competence of Oocytes Recovered Transvaginally from Holstein Cows for In vitro Fertilization

Background: Season of the year can affect the reproductive behavior in Holstein cows, altering the competition of the oocytes, reflecting a reduced production of embryos. The objective of this study was to evaluate the average of total oocytes, competition of oocytes and embryos in the in vitro production process at different season of the year in Holstein cows. Methods: During the four seasons of the year, was performed on each of the oocyte donor cows (winter, n = 957; spring, n = 1571; summer, n = 1776; autumn, n = 1128), by in vivo transvaginal follicular aspiration technique after the collection were subjected to the embryos production in vitro. Result: The highest number of total embryos were produced in winter and autumn, compared to spring and summer (3.76±0.16 and 3.54±0.18 vs. 2.73±0.11 and 2.45±0.10; respectively, P less than 0.05). During winter, a higher percentage of oocyte competition was observed, followed by autumn and spring and less competition shown in summer (26.03±0.39, 19.08±0.29, respectively, P less than 0.05). The quantity and competence of the oocytes collected and in vitro embryo production were drastically reduced during the hottest months of the year in this area of intense heat.

Analysis of IVF live birth outcomes with and without preimplantation genetic testing for aneuploidy (PGT-A): UK Human Fertilisation and Embryology Authority data collection 2016\u20132018

PurposeTo examine the live birth and other outcomes reported with and without preimplantation genetic testing for aneuploidy (PGT-A) in the United Kingdom (UK) Human Embryology and Fertilization Authority (HFEA) data collection.MethodsA retrospective cohort analysis was conducted following freedom of information (FoI) requests to the HFEA for the PGT-A and non-PGT-A cycle outcomes for 2016–2018. Statistical analysis of differences between PGT-A and non-PGT-A cycles was performed. Other than grouping by maternal age, no further confounders were controlled for; fresh and frozen transfers were included.ResultsOutcomes collected between 2016 and 2018 included total number of cycles, cycles with no embryo transfer, total number of embryos transferred, live birth rate (LBR) per embryo transferred and live birth rate per treatment cycle. Data was available for 2464 PGT-A out of a total 190,010 cycles. LBR per embryo transferred and LBR per treatment cycle (including cycles with no transfer) were significantly higher for all PGT-A vs non-PGT-A age groups (including under 35), with nearly all single embryo transfers (SET) after PGT-A (significantly more in non-PGT-A) and a reduced number of transfers per live birth particularly for cycles with maternal age over 40 years.ConclusionThe retrospective study provides strong evidence for the benefits of PGT-A in terms of live births per embryo transferred and per cycle started but is limited in terms of matching PGT-A and non-PGT-A cohorts (e.g. in future studies, other confounders could be controlled for). This data challenges the HFEA “red traffic light” guidance that states there is “no evidence that PGT-A is effective or safe” and hence suggests the statement be revisited in the light of this and other new data.

Randomized Controlled Study of the Effects of DHEA on the Outcome of IVF in Endometriosis.

Objective To investigate the effect of dehydroepiandrosterone (DHEA) on the outcome of in vitro fertilization (IVF) in patients with endometriosis (EMT). Methods Female patients diagnosed with EMT in our hospital from May 2018 to May 2019 were selected. The patients were divided into the control group (n = 22) and the DHEA group (n = 22) according to the random number table. Patients in the control group received placebo and patients in the DHEA group received DHEA. Patients in both groups received either DHEA (25 mg) or placebo orally 3 times a day for 90 days from the first day of menstruation. Patients were subsequently treated with an IVF cycle. In the control group, 22 patients completed the first cycle and 13 patients completed the second cycle. In the DHEA group, 22 patients completed the first cycle and 11 patients completed the second cycle. Serum sex hormone levels including serum E2 on hCG day, mean progesterone on hCG day, FSH on day 2, AMH on day 2, and gonadotropin dose were determined using a chemiluminescent immunoassay kit. The number of antral follicles of the bilateral ovaries was counted by transvaginal B-ultrasound, and the maximum length and transverse diameter of the ovaries were measured at the same time, to calculate the average diameter of the ovaries, observe the morphology of endometrium, and measure the thickness of the endometrium. The implantation rate, clinical pregnancy rate, persistent pregnancy rate, and live birth rate were compared between the two groups. Results There were no significant differences in serum E2, progesterone, endometrial thickness, recovered oocytes, mean number of transferred embryos, and mean score of leading embryo transfer between the DHEA group and the women who completed the first and second cycles (P > 0.05). The AMH, antral follicle count, serum E2 on hCG day, the number of recovered oocytes, fertilized oocytes, and the fertilization rate in the DHEA group were higher than those in the control group (P < 0.05). The doses of FSH on day 2, COH on day 3, and gonadotropin were lower than those in the control group (P < 0.05). There was no significant difference in the total number of embryos, the number of high-quality embryos, and the number of transplanted embryos between the two groups (P > 0.05). The implantation rate, clinical pregnancy rate, persistent pregnancy rate, and live birth rate in the DHEA group were higher than those in the control group (P < 0.05). Conclusion DHEA can significantly increase serum E2 level and improve IVF outcome by regulating the hormone synthesis process, thus improving oocyte and embryo quality.

Estimation of genetic parameters for superovulatory response traits in Japanese Black cows.

The aim of this study was to estimate genetic parameters for superovulatory response traits in order to explore the possibility of genetic improvement in Japanese Black cows. We analyzed 19 155 records of the total number of embryos and oocytes (TNE) and the number of good embryos (NGE) collected from 1532 donor cows between 2008 and 2018. A two-trait repeatability animal model analysis was performed for both. Because records of TNE and NGE did not follow a normal distribution, the records were analyzed following no, logarithmic, or Anscombe transformation. Without transformation, the heritability estimates were 0.26 for TNE and 0.17 for NGE. With logarithmic transformation, they were 0.22 for TNE and 0.18 for NGE. With Anscombe transformation, they were 0.26 for TNE and 0.18 for NGE. All analyses gave similar genetic correlations between TNE and NGE, ranging from 0.60 to 0.71. Spearman’s rank correlation coefficient between breeding values of cows with more than 10 records was ≥0.95 with both transformations. Thus, the genetic improvement of TNE and NGE of donor cows could be possible in Japanese Black cattle.

Оценка репродуктивной способности самцов крыс при остром токсоплазмозе

Toxoplasmosis is a parasitic disease of humans and animals caused by Toxoplasma gondii. Toxoplasma is an intracellular parasite that belongs to the simplest and has a complex development cycle. Infection with Toxoplasma is possible orally, transplacentally, percutaneously (if the integrity of the skin is damaged). This invasion is often the cause of problems with bearing pregnancy, as well as the development of congenital anomalies in children. The purpose of the study was to study the reproductive ability of male rats in acute toxoplasmosis. Materials and methods. The experiment was performed on 90 female and 45 male Wistar rats with a body weight of 180-200 g. The intact control males were orally injected with 2 ml of 0.2% starch gel. Experimental groups of males were infected with an invasive Toxoplasma gondii culture at a dose of 25 tachyzoites per 1 g of body weight (5000 tachyzoites per rat) and 50 tachyzoites per 1 g of body weight (10000 tachyzoites per rat). Then the males of all groups were coupled with the females for 3 days. The effect of toxoplasmas on the reproductive ability of male rats was assessed by the development of pregnancy and changes in the levels of pre- and post-implantation embryo death in female rats on the 7th, 14th, and 21st days after pregnancy. To account for changes in the pre- and post-implantation death of embryos in female rats after removal from the experiment, the uterus and ovaries were isolated, the uterine horns were opened, the number of implantation sites, the total number of embryos, the number of living and dead embryos, the number of resorption, and the number of yellow bodies in the ovaries were determined. Results and discussion. In the females of the 4th, 5th and 6th groups (coupling with males infected at the dose of 25 tachyzoites per 1 g of body weight), a decrease in the number of implantation sites in the uterus, the total number of embryos and the number of live embryos was recorded by 1.8-1.9 times compared to the control parameters. In female rats of the 7th, 8th and 9th groups (coupling with males infected at the dose of 50 tachyzoites per 1 g of body weight), there was a decrease in the number of implantation sites in the uterus, the total number of embryos and the number of live embryos by 5.6-6.8 times compared to the control. When compared to the results obtained from the females of the 4th, 5th and 6th groups, a decrease in these indicators was recorded by 3.1-3.5 times. Conclusion. Toxoplasma gondii has an effect on reproductive capacity in male rats expressed in changes of the levels of preimplantation mortality in female rats. The obtained effect depends on the dose of infection and the period of parasitosis development in males

MICROFLUIDIC SPERM SEPARATION DEVICE (MSSD) AND DENSITY GRADIENT CENTRIFUGATION (DGC): COMPARISON OF IMPROVEMENTS OF OUTCOMES USING PAIRED COMPARISONS

Comparison of a new technique to an older technique is often performed using patients who performed the new technique after they used old technique. Typically patients perform better in subsequent cycles, especially if they were unsuccessful in the first cycle. We chose to adjust this by comparing euploidy improvement within couples that underwent two consecutive IVF cycles utilizing either MSSD or DGC in the second cycle. Couples (N=150) undergoing a second cycle within a year period employing both ICSI and trophectoderm biopsy (TEBX) for preimplantation genetic testing for aneuploidy were reviewed for effects on outcome parameters based on sperm preparation techniques. All couples has DGC performed in first cycle were divided into two groups based on the sperm preparation technique in the second cycle. Group I (N=56); MSSD, Group II (N=94); DGC. T-test of (MSSD2-DGC1) VS. (DGC2-DGC1) compared MSSD with DGC. The t-test of paired comparisons showed there were no significant difference in maternal age, number mature eggs, 2PNs, number biopsied, number euploid or any of the stimulation parameter (not shown). Although there were no differences in total number of embryos biopsied, Group I, (MSSD) the proportions of biopsied embryos were significantly smaller on day 5 and larger on day 6 than DGC. Further, the post-processing motility was significantly higher in MSSD (improving from 74% to 94% MSSD) than with DGC (relatively unchanged from 72.5% to 70.6% with DGC in Group I).Table 1Comparisons of variables between MSSD and DGC (Paired comparison within groups)Mean Maternal ageMean # MIIMean # 2PNMean # biopsied% D5 BX%D6 BXMean # EuploidInitial Sperm motilityPost- processing sperm motilityGroup IDGC138.1 ± 3.88.5 ± 6.06.0 ± 3.93.3 ± 2.40.5 ± 0.40.4 ± 0.30.9 ± 1.146.6 ± 17.674.3 ± 25.6MSSD238.3 ± 3.79.3 ± 6.36.8 ± 5.13.5 ± 3.20.4 ± 0.30.5 ± 0.41.3 ± 1.944.3 ± 17.094.1 ± 8.9Δ MSSD2-DGC10.3 ± 0.70.8 ± 4.00.8 ± 3.70.2 ± 2.5-0.1 ± 0.50.1 ± 0.40.3 ± 1.5-2.3 ± 18.719.8 ± 26.6P value0.850.740.890.530.040.010.450.480.001Group IIΔDGC2-DGC10.3 ± 0.51.0 ± 4.80.9 ± 4.00.5 ± 2.70.01 ± 0.4-0.1 ± 0.50.5 ± 1.5-0.2 ± 17.5-1.9 ± 20.9DGC138.0 ± 4.08.5 ± 5.36.0 ± 4.43.2 ± 2.80.4 ± 0.40.5 ± 0.40.9 ± 1.342.6 ± 18.572.5 ± 26.6DGC238.3 ± 4.09.5 ± 6.06.9 ± 5.03.6 ± 3.30.5 ± 0.40.5 ± 0.41.5 ± 1.842.5 ± 19.770.6 ± 27.0 Open table in a new tab No effect was seen on outcome parameters based on sperm preparation technique used in this study. Subsequent IVF cycle showed trend to higher euploid rates irrespective of sperm preparation technique consistent with previous finding from our fertility center (Hodes-Wertz et al, 2015).

P–144 Undisturbed culture in time-lapse systems improves embryo development and quality

Abstract Study question Does culture in integrated time-lapse systems (TLS) improve embryo development and blastocyst quality compared to conventional benchtop incubators (CI), within the same IVF laboratory? Summary answer Under similar conditions, culture in TLS resulted in a significant increase in blastocyst rate, top quality blastocyst rate and proportion of biopsied embryos per treatment What is known already Integrated TLS have the potential of delivering a stable and undisturbed environment throughout the whole embryo culture, avoiding taking them out for assessment. However, there is still lack of quality evidence of the performance of these incubators compared to CI at supporting embryo culture until blastocyst stage. Studies abording this issue are still scarce, heterogeneous and have a small sample size. Although some authors have reported an improvement in embryo development and quality using TLS, global results are inconsistent. To our knowledge, the present study evaluates the effect of TLS on embryo quality on the largest sample size yet. Study design, size, duration Unicentric retrospective cohort study including 14248 ICSI treatments from 2016 to October 2020, with both autologous and donated oocytes. We compared blastocyst rate (BR) and proportion of top-quality blastocysts (TQB=Morphology ASEBIR score A) per treatment between those using TLS (N = 7500) and CI (N = 6748), and the proportion of embryos biopsied (EB) in cycles with pre-implantation genetic testing (PGT-A; N = 2642). We performed a sub-analysis in treatments using single-step culture medium (N-TLS=4398, N-CI=1140) in both types of incubators. Participants/materials, setting, methods Embryo cohorts were cultured until blastocyst stage in one of 3 TLS: EmbryoScope, EmbryoScope Plus (Vitrolife,) and Geri (Genea Biomedx), or in a CI (ASTEC). Embryo quality was assessed following ASEBIR morphological criteria. Culture protocols and media changed during the included time period. For that reason, we did a sub-study in the treatments performed since the implementation of Gems® (Genea Biomedx) single-step (SS) culture medium in all incubators. Statistical analysis was done using ANOVA tests. Main results and the role of chance Treatments were differently distributed and heterogeneous in terms of number of oocytes obtained per patient, so we stratified the analysis according to ovum origin and compared mean rates per cycle instead of total number of embryos per group. BR was statistically higher (P &amp;lt; 0,001) in the TLS group, in both autologous (62,98±29,37% vs 59,49±31,09% in CI) and oocyte donation treatments (69,25±22,07% vs 66,27±23,28% in CI). Proportion of TQB was also significantly higher in the TLS in both types of cycles (P &amp;lt; 0,001): 3,60±12,29% in TLS vs 2,27±9,71% in CI in autologous cycles, 8,68±15,31% in TLS vs 7,32±14,02% CI in ovum donation cycles. Results were corroborated in the SS media sub-study (P &amp;lt; 0,05): BR was 63,87±29,23% in TLS vs 57,53±30,61% in CI with autologous oocytes, and 70,76±21,63% in TLS vs 67,39±22,68% in CI with donated oocytes; TQB rates were 3,66±12,06% in TLS vs 2,05±9,26% in CI in autologous treatments and 8,81±15,21% in TLS vs 6,84±12,91% in CI in ovum donation treatments. Regarding PGT-A treatments, we found no significant difference in the biopsy rate in the total comparison, although the rate significantly increased in the TLS group since the implementation of single-step medium (52,36±24,69% in TLS vs 48,63±22,56% in CI; P = 0,007) Limitations, reasons for caution Not only culture conditions varied over time, but also the number of TLS in the laboratory, which increased lately. Hence, even though the most recent treatments included in the all-SS sub-study are more homogeneous in terms of culture conditions, they are unbalanced regarding the distribution among incubators. Wider implications of the findings: Our results demonstrate the superiority of TLS coupled with single-step culture media against traditional embryo culture systems at supporting embryo development. The optimal environment provided by TLS enhances embryo development until blastocyst stage as well as their quality, increasing the cumulative chances of getting a life-birth for each patien. Trial registration number Not applicable

VARIATION IN CYCLE CHARACTERISTICS AMONG PATIENTS < 35 WITH A SINGLE DIAGNOSIS

BackgroundInfertile women under 35 are among those considered to have a favorable prognosis with regards to assisted reproductive technology (ART) cycles. However, although reproductive age is the best predictor of outcome with ART, this population has a heterogeneous result with regards to live birth outcomes. Cycle characteristics, e.g. fertilization and embryo development, represent a potential counseling tool with which to manage patient expectations through an ART cycle. We hypothesized that infertility diagnosis would be associated with cycle characteristics among patients with ovarian (e.g. diminished ovarian reserve/DOR), anatomic (tubal factor/TF) and unexplained infertility.ObjectiveThe purpose of this study was to evaluate cycle characteristics among patients with DOR, TF, and unexplained infertility.Materials and MethodsA retrospective chart review of fresh autologous IVF cycles in patients < 35 years of age with a single diagnosis of unexplained infertility, tubal factor, or diminished ovarian reserve (DOR) performed at an academic center between 2010 and 2019 were identified. Patients with more than one identified indication for ART were excluded from analysis. Cycle characteristics were defined as total gonadotropin dose, number of oocytes retrieved, fertilization rate and total number of embryos for transfer and cryopreservation. ANOVA with post-hoc Tukey HSD was performed on variable outcomes.ResultsA total of 332 cycles were evaluated. In cycles with women < 35 years of age with a single diagnosis, unexplained infertility (46%; 153/332 cycles) was the most common single diagnosis, followed by DOR (36%; 119/332 cycles) and TF (18%, 60/332). No differences were observed in mean ages among unexplained (31.1 +/- 7.4 years), DOR (31.1 +/- 2.7 years), and TF (30.9 +/- 2.6 years) infertility (p=0.8). DOR cycles exhibited lower AMH values (1.5 +/- 1.62 ng/ml) versus unexplained infertility (3.27 +/- 1.84 ng/ml, p=0.002), higher total FSH dose (3801 +/- 1977 versus 2090 +/- 1005 mIU/mL, p<0.001), and lower total number of oocytes retrieved (9.3 +/- 5.3 versus 15.1 +/- 7.4, p<0.0001). No difference was observed in AMH, total FSH dose, and total number of oocytes retrieved between unexplained infertility and TF cycles. No statistically significant difference was seen in fertilization rates between the three groups (p=0.2); however, total number of embryos available for either transfer or cryopreservation was highest in tubal factor patients (7.1 +/- 6.6) compared to both unexplained (4.7 +/- 3.9, p<0.01) and DOR (3.7 +/- 3.0, p<0.001).ConclusionsDespite similar markers of ovarian reserve, total number of oocytes retrieved and fertilization rates, patients with unexplained infertility exhibit lower numbers of total embryos available for transfer or cryopreservation compared to those with TF infertility. These data suggest embryonic development as a potential contributor to ART outcomes in unexplained infertility.SUPPORT: N/A

Влияние хронического токсоплазмоза на изменение уровней пред- и постимплантационной гибели в эксперименте

The purpose of the study was to study the effect of chronic toxoplasmosis on changes in the levels of pre- and post-implantation mortality in the experiment. Materials and methods. In the experiment there were 90 female Wistar rats with a body weight of 180-200 g. For the development of pregnancy, females of the control and experimental groups were coupled with males for 3 days. After the onset of pregnancy, females of intact controls were orally injected with 2 ml of 0.2 % starch gel. The females of the experimental groups were infected with an invasive culture of Toxoplasma gondii at a dose of 25 tachyzoites per 1 g of body weight (5,000 tachyzoites per rat) and 50 tachyzoites per 1 g of body weight (10,000 tachyzoites per rat). On the 35th day after infection, females of the experimental groups were coupled with males for 3 days before infection. The effect of Toxoplasma on changes in the levels of pre- and post-implantation death was assessed after killing female rats on the 42nd, 49th and 56th (7th, 14th and 21st days after pregnancy) days after infection. Results and discussion. In animals infected at a dose of 25 tachyzoites per 1 g of body weight, there is a decrease in the number of implantation sites in the uterus, the total number of embryos and the number of living embryos at all stages of the development of Toxoplasma by 1.8-2.1 times compared with the control. In females infected at a dose of 50 tachyzoites per 1 g of body weight, a decrease in the number of implantation sites in the uterus and the total number of embryos at all stages of the development of the parasite was recorded by 2.2-2.5 times compared with the control values. There is a decrease in the number of living embryos by the 42nd day after infection by 4.3 times, by the 49th day – by 3.8 times and by the 56th day – by 5.1 times compared to the control. When compared with the results obtained from females with a lower dose of infection, a decrease in this indicator was revealed on the 42nd day after the development of Toxoplasma by 2.1 times, on the 49th day – by 1.7 times and on the 56th day – by 2.5 times. An increase in the number of dead embryos by 0.2-0.4 times in comparison with intact indicators and the results of experimental animals infected at a dose of 5,000 tachyzoites per rat were recorded. There was an increase in the level of resorptions on the 49th and 56th days by 1.4 and 1.6 times in comparison with the control and animals infected at a dose of 25 tachyzoites per 1 g of body weight. Conclusion. Experimental chronic toxoplasmosis causes an increase in pre-implantation and post-implantation embryo death. The recorded effect of Toxoplasma invasion depends on the dose of infection and the duration of the disease

COMPARATIVE ANALYSIS OF THE MORPHOLOGICAL CONDITION OF IN VIVO OBTAINS OF EMBRYOS FROM DONOR COWS OF DIFFERENT BREEDS

The aim of the study was a comparative analysis of the morphological status of embryos of cattle obtained in vivo from purebred and local donor cows of the I and II generations.&#x0D; The object of experimental studies were obtained embryos of cattle in vivo. They were obtained in the experimental farm "Ukrainka" of the Institute of Animal Science of the UAAS of Ukraine from 1985 to 1990. Information about animals and the results of research are stored in the archives of the Private Company "Bioservice". In order to study the stages of development of seven-day-old embryos of cattle, the embryo productivity of 11 purebred and 72 local donor cows was analyzed. Purebred cows were Ayshir (4 heads), Simmental (1 head) and Ukrainian Black-and-White dairy (6 heads) breeds. Crossbreeds of the first generation were donor cows from crossing red steppe and Holstein (2 goals), Simmental and Holstein (12 goals) and Ukrainian Black-and-White dairy and Holstein (9 goals) breeds. The crossbreeds of the second generation were donor cows from crossing red steppe and Holstein (14 goals), Simmental and Montbeliard (22 goals) and Ukrainian Black-and-White dairy and Holstein (13 goals) breeds.&#x0D; According to the results of the conducted research, a statistically significant difference (p &lt; 0.01) between purebred (7.8%) and local donor cows of the I and II generation (0.6 and 1.4%, respectively) was established by the number of embryos. obtained at the stage of expanded blastocyst. There was also a statistically significant difference (p &lt; 0.001) at the stage of late blastocyst between purebred (38.0%) and local donor cows of the I and II generation (7.1 and 9.2%, respectively). The analysis of the obtained research results shows that from purebred donor cows a significantly larger (p &lt; 0.001) total number of blastocysts was obtained by 60.4% compared to local donor cows of the I and II generation (19.6 and 24.2%, respectively).&#x0D; It was found that at the stage of late morula found significantly more embryos 44.7% and 32.9 (p &lt; 0.01), respectively, in donor cows of the second and fourth generation compared to purebred 19.4%. It should be noted that at the stage of early morula, a significantly larger number of embryos (p &lt; 0.001) was obtained from donor cows of the I and II generation (13.6 and 3.2%, respectively) compared to purebred 0.0%. According to the results of the comparative analysis of the total number of obtained morulae, a significantly higher number of embryos (p &lt; 0.001) obtained from donor cows of the I and II generation (48.6 and 50.0%, respectively) was found in comparison with purebred 21.7%.&#x0D; It was found that purebred donor cows received a greater 82.2% of the total number of suitable embryos compared to 68.2% (p &lt; 0.001) of local donor cows of the first and 73.5% (p &lt; 0.05) from local donor cows of the second generations.&#x0D; Further development of extracted morulae in vitro from purebred and local donor cows on the seventh day at a temperature of 37.5°C for 15 hours led to the formation of full-fledged blastocysts at 73.0% (400 of 548).&#x0D; It was found that purebred donor cows received more blastocysts (p &lt; 0.001) compared to local donor cows of the first generation by 40.8% and 36.2% compared to local donor cows of the second generation.&#x0D; It was found that from local donor cows of the second generation received 28.3%, and from local donor cows of the first generation by 26.9% more (p &lt; 0.001) morula compared to purebred donor cows.&#x0D; According to the results of the research, it was found that purebred donor cows received a higher total number of embryos (molula + blastocyst) compared to local donor cows of the first generation by 14.0% (p &lt; 0.001) and 8.7% &lt; 0.05) compared with local donor cows of the second generation.

287 Effects of Number of Sperm and Site of Uterine Semen Deposition on Conception Rate and Number of Embryos in Weaned Sows Receiving a Single Fixed Time Insemination

Abstract Induced ovulation with single fixed time artificial insemination (SFTAI), combined with uterine (IUI) or deep uterine insemination (DUI), could improve fertility with low numbers of sperm and allow greater use of high genetic merit boars. At weaning (0 h), sows (n = 534) were assigned by parity and estrus induction method (eCG or Control) to receive 1200 × 106 sperm by IUI, 600, 300, or 150 × 106 sperm by IUI or DUI, or 75 × 106 sperm by DUI. At 80 h post weaning, sows received OvuGel and 26 h later a pooled semen SFTAI. Ultrasound was performed to determine follicle size and time of ovulation (OV). Sows were slaughtered 27 d after AI to determine pregnancy and litter traits. There was no effect (P &amp;gt; 0.05) of eCG on estrus (93%) within 5 d of weaning or follicle size (6.1 mm) at OvuGel, but wean-to-estrus (3.8 vs. 4.0 d,P &amp;lt; 0.01) and AI-to-OV (15.9 vs. 17.0 h, P = 0.04) intervals and AI-to OV were slightly reduced. eCG did not affect pregnancy rate (78.6%), number of CL (21.7), or number of viable embryos (12.2). There was no effect of number of sperm or site of insemination (P &amp;gt; 0.05) on pregnancy rate (range: 80.9% to 70.5%), but AI occurring after ovulation reduced pregnancy rate (P &amp;lt; 0.02). Total number of embryos (range: 16.5 to 10.3) increased with CL number (P &amp;lt; 0.001) but was not affected by number of sperm or site of insemination (P &amp;gt; 0.05). Higher sperm treatments (1200 and 600 x 106) had more embryos compared to lower sperm treatments (P &amp;lt; 0.01), suggesting that lower sperm numbers effects litter size more than the pregnancy status. Acceptable fertility can be achieved with low sperm numbers when using SFTAI and uterine deposition, but AI-to-OV interval and ovulation rate influence final fertility.

Embryo blastulation and quality between days 5 and 6 of extended embryo culture.

This study aims to know what proportion of culture day 5 pre-blastocyst-stage embryos develop into blastocysts by culture day 6 and what patient and cycle characteristics are associated with delayed blastocyst formation. A retrospective observational cohort analysis was performed including a total of 9886 embryos from 1008 IVF cycles in 835 patients, who underwent treatment between January 1, 2016, and December 31, 2018. Autologous fresh in vitro fertilization (IVF) cycles at a single academic center were included in the analysis. Embryos were group-cultured using single-step culture media. Blastulation was defined as the presence of a new blastocyst. Usable blastulation was defined as the presence of a new good or excellent quality, expanded, hatching, or hatched blastocysts. The mean blastulation rate between days 5 and 6 of extended embryo culture was 30.9%. The mean percentage of embryos developing into usable blastocyst-stage embryos was 19.8%. The factors associated with blastulation on day 6 included the total number of embryos and the number of pre-blastocysts on day 5, as well as the use of ICSI. Age, the number of total embryos, those remained in culture and pre-blastocysts, as well as the blastulation rate on day 5 were associated with usable blastulation. It is important to know the usable blastocyst development rate between culture days 5 and 6 in order to adequately counsel patients debating whether to proceed with fresh ET on day 5 or forego ET with the expectation that embryos will be biopsied for PGT and/or cryopreserved on culture day 6. Our findings provide evidence to help guide patients in this difficult decision.

Duration of stimulation in patients with polycystic ovarian syndrome undergoing ICSI: Does it affect the outcome?

Back ground: Oocyte and embryo quality are affected by the duration of ovarian stimulation in differentcategories of patients undergoing ICSI specially forthose having polycystic ovarian syndrome as theyundergo ovarian stimulation usinglow dose step up antagonist protocol.Methods: Retrospective analysis of 139 patient underwent ovarian stimulatiaon using antagonist protocol and freeze all policyResults: Patients were divided into 3 groups according to the duration of ovarian stimulation (group A≤ 8 days, group B 9-10 days, group C ≥ 11 days).We found that in spite of having no statistically significant difference in the total number of retrievedoocytes, there was statistically significant differencein the median number of germinal vesicle oocytesbetween group A and group C (3 (1-13) & 1(0-5) Pvalue= 0.01). Also this difference is present betweengroup B and group C (3 (1-11) & 1(0-5) P value=0.01). For metaphase II oocytes there was a statistically significant difference in the median numberbetween group A and group B (3 (0-8) & 5 (1-14)P value= 0.001). Also this difference is present between group A and group C (3 (0-8) & 3 (3-12) Pvalue= 0.001).There was no statistically significantdifference in the total number of embryos in betweenthe 3 groups.Conclusion: We found that it seems to be safe toextend the duration of stimulation more than 8 dayswhen treating patients with polycystic ovarian syndrome undergoing IVF/ICSI, as it was associatedwith more increase in mature oocytes (MII)

146 Single injection of follicle-stimulating hormone before ovum pickup in lactating Holstein donors: Oocyte recovery and embryo production

Lactating donor cows frequently have decreased oocyte quality, lower fertilization rates, and impaired early embryonic development due to their lactational metabolic challenges. Synchronization and stimulation of follicular growth before ovum pickup (OPU) has been used to improve oocyte quality and consequently, embryo production. The objective of this study was to evaluate the effects of a single injection of FSH before OPU on oocyte recovery and embryo production in lactating Holstein donors. During June and July 2020, 22 lactating Holstein donors (open, 40 to 90 DIM, producing &amp;gt;90 lbs of milk) from Ruann Dairy (Riverdale, CA) were randomly assigned to one of two treatments (crossover design). Donors did not receive any injections before OPU when assigned to the No FSH treatment. Treatment 1×FSH consisted of a single intramuscular (IM) injection of 140mg of FSH (Folltropin®, Vetoquinol) 36h after gonadotrophin-releasing hormone (GnRH; Fertagyl, Merck®, 129µg, IM). The FSH consisted of a 3.5-mL IM injection of 400mg of FSH diluted in 10mL of 0.5% hyaluronan (HA). OPU was performed 48 to 52h after FSH. All donors received both treatments on a 14-day interval. The recovered oocytes were fertilized with the same sexed female-sorted semen in both rounds (3 different sires were used). OPU, oocyte classification, IVM, IVF, and culture (IVC) were performed as described by Demetrio et al. (2020 Anim. Reprod. 17, e20200053). All oocytes went into IVM, except for degenerated occytes. The number of 4-cell (or more) embryos on Day 3 of IVC divided by the number of oocytes in IVC after IVF is defined as the cleavage rate. The number of blastocysts (early to hatched) on Day 7 of IVP divided by the number of oocytes in IVC after IVF is defined as the blastocyst rate. Poisson-normal (count data) and Logistic-normal (proportion data) models were used to analyse the data. Treatment, donor (random effect), and sire were included in the models. The results are summarised in Table 1. Oocyte recovery and embryo production were highly donor dependent. There were no differences in the number of recovered oocytes among treatments. Stimulation of the follicular growth before OPU with one single injection of FSH diluted in 0.5% HA 36h after GnRH improved oocyte quality, cleavage rates, blastocyst rates, embryo quality, and the total number of embryos per OPU in lactating Holstein donors. Table 1. Oocyte recovery, cleavage rate, and embryo production results Treatment OPU Oocytes per donor Grade 1 and 2 oocytes Cleavage rate (%) Blastocyst rate (%) Total embryos per OPU Grade 1 embryos per OPU No FSH 22 18.2 30a 69a 22a 3.4a 1.6a 1×FSH 22 17.5 34b 77b 34b 5.3b 3.3b a,bValues with different superscripts in the same column differ (at least P&amp;lt;0.05).

Factors Associated with In Vitro Fertilization Live Birth Outcome: A Comparison of Different Classification Methods.

BackgroundIn vitro fertilization (IVF) is a useful assisted reproductive technology to achieve pregnancy in infertile couples. However, it is very important to optimize the success rate after IVF by controlling for its influencing factors. This study aims to classify successful deliveries after IVF according to couples’ characteristics and available data on oocytes, sperm, and embryos using several classification methods.Materials and MethodsThis historical cohort study was conducted in a referral infertility centre located in Tehran, Iran. The patients’ demographic and clinical variables for 6071 cycles during March 21, 2011 to March 20, 2014 were collected. We used six different machine learning approaches including support vector machine (SVM), extreme gradient boosting (XGBoost), logistic regression (LR), random forest (RF), naïve Bayes (NB), and linear discriminant analysis (LDA) to predict successful delivery. The results of the performed methods were compared using accuracy tools.ResultsThe rate of successful delivery was 81.2% among 4930 cycles. The total accuracy of the results exposed RF had the best performance among the six approaches (ACC=0.81). Regarding the importance of variables, total number of embryos, number of injected oocytes, cause of infertility, female age, and polycystic ovary syndrome (PCOS) were the most important factors predicting successful delivery.ConclusionA successful delivery following IVF in infertile individuals is considerably affected by the number of embryos, number of injected oocytes, cause of infertility, female age, and PCOS.

73 Use of embryo transfer technology to salvage the germplasm of elite dromedary camels (Camelus dromedarius) infected with brucellosis

The present study was conducted to evaluate the potential of embryo transfer technology to salvage invivo-produced embryos from elite dromedary camels infected with brucellosis without transmission of the pathogen to the recipients or offspring. Ten elite racing champions who tested positive for brucella comprised the experimental group, whereas 3 donors negative for brucella acted as the control. Each donor animal received a combination of 2500IU of equine chorionic gonadotrophin (eCG; Folligon; Intervet), given as a single intramuscular injection on Day 1 of the treatment protocol, and 400mg of pFSH (Folltropin; Bioniche), injected twice daily in declining doses of 2×80mg, 2×60mg, 2×40mg, and 2×20mg over 4 days, also beginning on Day 1. They were mated with a fertile bull on the 11th day after the start of treatment. The donors of the group 1 received a combination of tetracycline, and streptomycin on alternate days for 2 weeks before mating with a fertile male. An injection of 20µg of gonadotrophin-releasing hormone (Receptal, Intervet) was given to them immediately after mating to induce ovulation. The embryos were collected by the non-surgical method on Day 7 day after ovulation as per the guidelines published by IETS. All embryos were washed at least 3 times in holding medium supplemented with antibiotics and then transferred individually into the left uterine horn of synchronized recipients. All recipients and calves born were tested for brucellosis every 3 weeks. The data were analysed using a two-sample t-test (Minitab statistical software, Minitab Ltd.). Results are shown in Table 1. No difference was observed in the number of embryos collected per flushing and pregnancies established at Day 60. The proportion of pregnancies reaching term from the total number of embryos transferred and from the pregnancies established on Day 60 did not differ from embryos obtained from brucella-infected and control donors. All recipients tested negative for brucellosis during their gestation and until weaning. All calves born were also negative for brucellosis on birth and until weaning and handing over to clients. In conclusion, this is the first study in camels wherein we have demonstrated that invivo-produced embryos from elite dromedary females infected with brucellosis could be collected and transferred to synchronized recipients without transmitting the pathogen to the recipients or offspring. Table 1. Pregnancies established from invivo-produced embryos collected from elite dromedary camels infected with brucellosis Donor group (N) Total number of flushings No. of embryos collected per flushing (n±s.e.m.) Pregnancies on Day 60 (% mean±s.e.m.) Pregnancies reaching term (% mean±s.e.m.) from: Embryos transferred Pregnancies on Day 60 Brucella (10) 30 5.4±0.6 55.1±2.6 44.4±2.3 83.2±3.6 Control (3) 6 5.3±0.6 65.7±6.3 53.3±6.8 80.9±7.7 P-value 0.969 0.173 0.262 0.797

The Predictive Value of the Follicular Output Rate on Pregnancy Outcome of Patients with Polycystic Ovary Syndrome Undergoing In Vitro Fertilization and Embryo Transfer.

BackgroundThis retrospective study aimed to evaluate the predictive value of the follicular output rate (FORT) on the pregnancy outcome of patients with polycystic ovary syndrome (PCOS) undergoing in vitro fertilization and embryo transfer (IVF-ET).Material/MethodsBetween January 2012 and June 2016, a total of 1,541 patients with PCOS who underwent IVF-ET at our center were enrolled in the study. FORT was calculated as the pre-ovulatory follicle count (PFC)/antral follicle count (AFC)×100%.ResultsAccording to the FORT, patients were divided into low, medium, and high FORT groups. With an increase in the FORT, the PFC and serum estradiol at the day of human chorionic gonadotropin (hCG) injection, the number of retrieved oocytes, metaphase II (MII) oocytes, total number of embryos, and number of high-quality embryos significantly increased (P<0.05 and P<0.001) from the low to high FORT groups, while the AFC, gonadotropin (Gn) stimulation day, and total Gn decreased significantly (P<0.001). The live birth rate from frozen embryo transfer and the cumulative live birth rate was the lowest in middle FORT group but increased significantly in high FORT group (P<0.05). The correlation analysis between FORT and related factors showed that the FORT was negatively correlated with body mass index (BMI), Gn stimulation days, and total Gn (P<0.05).ConclusionsFORT is a powerful tool for measuring ovarian reactivity. For patients with PCOS, a high FORT to obtain high-quality embryos and perform frozen embryo transplantation can achieve good pregnancy outcome.

Comparative study between ovarian response prediction index versus anti-Müllerian hormone, antral follicular count as predictors of ovarian response in women undergoing IVF/ICSI cycles

Aim:This study aimed to compare individual predictors as AMH, AFC versus the ovarian response prediction index in determining the ovarian response to controlled ovarian stimulationMaterials and Methods: A prospective cohort study conducted on 84 infertile couples candidate for Intracytoplasmic sperm injection (ICSI) for the first time using either long agonist or antagonist protocol according to clinical evaluation. Participants were classified according to the number of MII oocytes into poor response with 3 or less oocytes and normal responders with 4 or more oocytes.Results: The study showed that women with poor response were statistically older than those with normal ovarian response (33.1±5.9 vs. 29.8±5.4, respectively). The number of cumulus (12.1±5.2 vs. 2.5±1.5), MII oocytes (7.8±3.6 vs. 2±0.8), grade A embryos (3±0.8 vs. 1.4±0.9) and total number of embryos (3.8±2.2 vs. 1.7±0.7) were significantly higher in normal responders. ORPI has the highest accuracy in predicting ovarian response (88%) when compared to AMH (83%) and AFC (86%).Conclusion: AMH, AFC and ORPI are good predictive of the ovarian response and help in choosing the protocol and gonadotropin dose of induction and prediction of OHSS