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PubMed1989

Delayed fertilization during in vitro fertilization and embryo transfer cycles: analysis of causes and impact on overall results.

This study evaluated possible causes of delayed fertilization during in vitro fertilization (IVF) cycles, its repetitiveness, and its influence on IVF results in 23 patients (27 cycles) with delayed fertilization of greater than or equal to 1 preovulatory oocyte(s). In 15 cycles, reinsemination with husband's semen was performed at 18 hours. Possible causes of delayed fertilization were oocyte defects (10 cycles, 37.0%), sperm defects (4 cycles, 14.8%), oocyte and sperm defects (4 cycles, 14.8%), and no detectable gamete defects (9 cycles, 33.3%). Overall fertilization rate was 47.9%. No pregnancies were observed in 10 patients with one embryo transferred. Recurrence rate of delayed fertilization per patient was 17.3%; overall ongoing pregnancy rate/cycle was 10.3%. Although repetitiveness of delayed fertilization is low, it seems to impact negatively on IVF results.

Embryo Transfer↗
PubMed1996

Serum from women with polycystic ovary syndrome inhibits fertilization and embryonic development in the murine in vitro fertilization model.

OBJECTIVE: To study the effects of serum collected from women with polycystic ovary syndrome (PCOS) on the fertilization and early embryonic development of the murine oocyte. DESIGN: Sera from women with anovulation were used as a supplement in a murine IVF model. SETTING: Tertiary care academic medical center. PATIENTS: Four fertile, four hypothalamic amenorrheic, seven PCOS (three with elevated LH and four with elevated T), and three anovulatory women with normal hormone levels. RESULTS: When compared with serum from fertile women, serum from women with PCOS reduced fertilization rates (60% versus 42%) and subsequent early embryonic development rates (87% versus 31%). Serum from women with PCOS and elevated T levels had the lowest fertilization rates (22%). Polycystic ovary syndrome serum with elevated T or LH levels significantly decreased early embryonic development rates in comparison to fertile women (22%, 41% versus 87%). CONCLUSIONS: Serum from women with PCOS inhibited fertilization and blastocyst development. Because both T and LH caused inhibited blastocyst development, these data have implications regarding low pregnancy rates and live birth rates during ovulation induction in women with anovulation. These data also raise questions regarding the use of serum during IVF.

Amenorrhea↗
PubMed1989

Fertilization and cleavage of mouse oocytes exposed to the conditions of human oocyte retrieval for in vitro fertilization.

Ova from two strains of mice (a hybrid-inbred strain, B6D2F1, and a random-bred strain, CD1) were shocked by exposure to environmental conditions possibly encountered by human oocytes retrieved for in vitro fertilization (IVF). Shocked and control mouse ova were fertilized in vitro in either simple or complex media and zygote development to morulae and blastocyst stages compared with that of zygotes fertilized in vivo. Development of the hybrid-inbred zygotes following fertilization in the simple media of shocked and control ova was essentially the same as for ova fertilized in vivo (84 +/- 6.6, 89 +/- 1.6, 87 +/- 6.0% to the 2-cell stage and 89 +/- 5.2, 94 +/- 2.3, 99 +/- 1.0% of two cells to blastocysts, respectively); development in the complex media also was the same following fertilization of shocked and control ova (80 +/- 8.7, 90 +/- 2.7% to two cells and 33 +/- 3.5, 35 +/- 4.5% of two cells to blastocysts, respectively) but lower than that of in vivo zygotes (92 +/- 4.6 to two cells, 58 +/- 4.7 two cells to blastocysts). In contrast, the fertilization and development in the simple media of shocked and control random-bred ova was lower and more variable (90 +/- 5.8, 67 +/- 11.1% to two cells and 17 +/- 8.3, 42 +/- 13.6% two cells to blastocysts, respectively) than the development of in vivo zygotes (96 +/- 1.5% to two cells, 51 +/- 5.5 two cells to blastocysts).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2003

Effects of oviductal and cumulus cells on in vitro fertilization and embryo development of porcine oocytes fertilized with epididymal spermatozoa.

This study was designed to evaluate the effects of adding porcine oviductal epithelial cell (POEC) monolayers before or during the fertilization of denuded or cumulus-enclosed oocytes, in terms of fertilization results and subsequent embryo development. The variables determined were: penetration rate, mean number of spermatozoa per oocyte, male pronucleus formation rate, monospermy rate, cleavage rate after 48 h of fertilization, blastocyst rate, and mean number of nuclei per blastocyst. We used cumulus-free and cumulus-enclosed oocytes preincubated or fertilized in the presence of POEC, once the purity in epithelial cells of these cultures had been assessed. All the experiments involved the use of frozen-thawed epididymal spermatozoa to avoid replicate variability. The POEC cultures prepared showed a high proportion of epithelial cells (over 95%). Preincubation of oocytes with POEC before fertilization showed no effects on the fertilization variables determined. In contrast, during IVF under our experimental conditions, these cells attached to the cumulus cells and their interaction had a significant effect on some of the fertilization variables analyzed. The presence of POEC and cumulus cells during IVF increased oocyte penetrability. Moreover, in the absence of POEC, cumulus cells resulted in a reduced monospermy rate. On subsequent embryo culture, a lower cleavage and blastocyst formation rate were recorded when the oocytes had been preincubated with POEC before IVF.

Animals↗
PubMed1999

Fertilization, embryo quality, and cryosurvival in in vitro fertilization and intracytoplasmic sperm injection cycles.

PURPOSE: Our purpose was to investigate the influence of semen quality on fertilization, embryo morphology, cleavage, and cryosurvival in conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) programs. METHODS: A retrospective analysis of 513 couples undergoing IVF and 255 couples undergoing ICSI was done. RESULTS: Semen quality influenced fertilization in IVF and abnormal fertilization in IVF and ICSI, but no effects on the development, morphology, implantation capacity, or cryosurvival of embryos were found. Fertilization, embryo quality, and cryosurvival rates were similar after IVF and ICSI. The fertilization rate of mature oocytes in IVF was lower when cytoplasmic immaturity in the oocyte population was frequent. The speed of development of embryos was 2 hr faster after ICSI than after IVF. Two-cell-stage embryos survived best after cryopreservation with propanediol and sucrose on day 2. CONCLUSIONS: After fertilization, semen parameters had no effect on the quality or cryosurvival of embryos in either IVF or ICSI.

Adult↗
PubMed2003

Seminal reactive oxygen species as predictors of fertilization, embryo quality and pregnancy rates after conventional in vitro fertilization and intracytoplasmic sperm injection.

High seminal reactive oxygen species (ROS) are related to poor semen quality and impaired fertilization. We aimed at finding whether there is an association between ROS and fertilization, embryo quality and pregnancy rates after conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). In prepared semen of 147 male partners of infertile couples, ROS were assessed with luminol chemiluminescence. Spermiogram was assessed in native semen. ROS were negatively correlated with standard sperm characteristics and testicular volume, and positively with abnormal sperm head morphology. Fertilization rate and embryo morphology on day 2 and on day 4 were assessed in 41 IVF and 106 ICSI cycles. The influence of maternal (female age and number of oocytes) and paternal (sperm motility, morphology and ROS) factors on fertilization and embryo quality were assessed by means of regression analyses. After IVF, fertilization and pregnancy rates were negatively associated with ROS level (p = 0.031 and 0.041, respectively). In case of higher ROS, significantly fewer ICSI-derived embryos (p = 0.036) reached the morula-blastocyst stage on day 4. High seminal ROS levels are associated with impaired sperm fertilizing ability and lower pregnancy rates after IVF. In ICSI, a negative association of ROS with embryo development to the blastocyst stage has been observed.

Adult↗
PubMed1995

Assessment of fertilization failure and abnormal fertilization after intracytoplasmic sperm injection (ICSI).

The assessment of fertilization is an important part of intracytoplasmic sperm injection (ICSI) and oocytes are routinely examined about 17 h after injection using Nomarski differential interference contrast optics. However, it is not possible to conclusively determine the aetiology of fertilization anomalies in this manner, so cytological studies were undertaken to determine the causes of failed and abnormal fertilization after ICSI. Oocytes which exhibited no evidence of fertilization, one pronucleus (PN) or 3 PN were fixed in glutaraldehyde, stained with Hoechst 33342 and examined by fluorescence microscopy to identify PN, metaphase chromosomes, sperm heads and polar bodies. A total of 428 unfertilized oocytes were examined from 170 ICSI cycles. Overall, 82% of these unfertilized oocytes were still at metaphase II (non-activated) while the remaining 18% were activated and had 1 PN and two polar bodies. The majority (71%) of the metaphase II oocytes contained a swollen sperm head, which indicates that the spermatozoon was correctly injected but the oocyte did not activate and complete its second meiotic division. The swollen sperm head was located among the metaphase chromosomes in 4.3% of these oocytes, while in some cases (6.6%), the sperm chromosomes had undergone premature chromosome condensation (PCC). Other aetiologies of failed fertilization in these metaphase oocytes were ejection of the spermatozoon from the oocyte (19%) and complete failure of sperm head decondensation (10%). A similar pattern of anomalies was found in 1 PN oocytes, although the ratios were different (swollen sperm head, 51%; ejection of the spermatozoon, 19%; undecondensed sperm head, 30%). Seventy abnormally fertilized oocytes were also examined, of which 63 had 3 PN and a single polar body, indicating that the unextruded second polar body developed into the third PN. In conclusion, the present study demonstrates that the principal cause of fertilization failure after ICSI is failure of oocyte activation and not ejection of the spermatozoon from the oocyte. It is also apparent that further studies are needed to elucidate the mechanisms that control oocyte activation and sperm head decondensation in injected oocytes.

Adult↗
PubMed1999

Sperm treatment with extracellular ATP increases fertilization rates in in-vitro fertilization for male factor infertility.

Previous work from our laboratory has revealed that extracellular ATP is a rapid and potent activator of human sperm acrosome reaction and fertilizing ability. In the present study, we assessed the effects of in-vitro sperm incubation with ATP on fertilization and embryo development in couples undergoing in-vitro fertilization (IVF) for male factor infertility. Oocytes from 22 women undergoing ovulation induction were divided in two groups and inseminated in vitro either with selected spermatozoa from the corresponding partner suffering from male factor infertility pre-incubated with ATP (2.5 mM) for 1 h, or with spermatozoa incubated with 0.9% NaCl solution (control group). After insemination, fertilization was assessed by the presence of pronuclei and then by embryo cleavage. The fertilization rate in the group of oocytes inseminated with ATP-treated spermatozoa improved significantly with respect to the control group (65.7 versus 42.5%, P < 0.01). No significant differences were observed in embryo cleavage and embryo quality. Embryos from both treated and control groups were transferred together in 20 transfer procedures, and in two couples fertilization was not obtained. Nine pregnancies occurred: one biochemical, one miscarriage, and seven patients delivered 9 healthy babies. Two pregnancies were twin with an overall pregnancy rate of 40.9% per cycle and of 45% per transfer. In conclusion, the results of the present study demonstrate that, in humans, extracellular ATP induces a significant increase of sperm fertilizing potential, as these findings are a rationale for the use of ATP for in-vitro treatment of human spermatozoa during IVF.

Adenosine Triphosphate↗
PubMed1995

Micromorphometry and spermatozoa binding patterns of fertilized and unfertilized human oocytes after in-vitro fertilization.

Human oocytes (n = 380) from 71 in-vitro fertilization patients were measured 18 h after insemination to find out if certain parameters of oocyte morphology could be related to fertilization. In addition, the number and distribution patterns of spermatozoa bound to or within the zona pellucida of 534 oocytes were analysed. The mean diameter of the human oocyte was 167.7 +/- 9.5 microns and its mean volume was 2.5 x 10(6) micron 3. There were no significant differences in diameter between 112 fertilized and 168 unfertilized oocytes, although they displayed differences in the size of the perivitelline space and the zona pellucida. The age of the patients had no significant effect on the morphometry of the oocytes. Sperm binding patterns did not correlate with fertilization. The number and distribution of the spermatozoa on the surface of the zona pellucida was extremely heterogeneous and was not related to the occurrence of fertilization. All possible binding and distribution patterns were in the same range in both fertilized and unfertilized oocytes. In conclusion, the micromorphometry of human oocytes and their sperm binding patterns were not related to the occurrence of fertilization.

Adult↗
PubMed1995

A comparative prospective study using matched samples to determine the influence of subnormal hypo-osmotic test scores of spermatozoa on subsequent fertilization and pregnancy rates following in-vitro fertilization.

The achievement of pregnancies in vivo is rare in couples where the male partner has defective sperm membranes as shown by hypo-osmotic swelling (HOS) test scores of < 50%. However, there have been mixed reports on the value of the HOS test in predicting outcome following invitro fertilization; some studies suggest reduced fertilization rates and others find little, if any, predictability of decreased fertilization. The assumption has been made that fertilization rates are proportional to pregnancy rates; however, this may not necessarily be true since defective spermatozoa could lead to a less viable pre-embryo and therefore a decreased viable pregnancy rate. We performed a comparative prospective study using matched controls to evaluate fertilization rates and to determine subsequent pregnancy rates. The mean HOS scores were 70.0 and 36.7% respectively, with mean motile sperm concentrations of 35.7 and 34.0 x 10(6)/ml in 27 matched pairs. There was no difference in the mean number of oocytes retrieved, fertilization rates or number of embryos transferred between the two groups by HOS score. The clinical and viable pregnancy rates and implantation rates were 25.9, 18.5 and 9.9% for normal versus 3.7, 3.7 and 1.1% for subnormal groups. These data suggest that low HOS scores may be associated with the formation of defective embryos, leading to low pregnancy rates but normal fertilization rates.

Female↗
PubMed1993

The ability of the hemizona assay to predict human fertilization in different and consecutive in-vitro fertilization cycles.

The objective of this prospective study was to examine the ability of the hemizona assay (HZA) to predict fertilization outcome of mature, pre-ovulatory oocytes under in-vitro fertilization (IVF) conditions. Since a large number of patients were evaluated over a long period, the power of the HZA to prognosticate fertilization results in the same and subsequent (consecutive) IVF cycles of those same patients was assessed. For IVF, only metaphase II oocytes were used. For the HZA, both fresh oocytes donated by patients at the time of IVF and oocytes recovered from surgically removed ovarian tissue (and salt-stored) were used, and bisected by micromanipulation techniques. Matching hemizonae were co-incubated either with spermatozoa from the patient (test) or from a fertile man (control) for 4 h. The number of spermatozoa tightly bound to the zona was counted. Patients (n = 112) were divided into two groups based on HZA results (expressed as HZA index or HZI): HZI > or = 30% (n = 72) and < 30% (n = 40). The patients with HZI < 30% had significantly lower fertilization rates in both the HZA-IVF cycle and in subsequent cycles compared to patients with HZI > or = 30% (P < 0.03). Linear discriminant analysis indicated the HZA to have a sensitivity of 84%, and positive and negative predictive values of 85 and 70% respectively, for prediction of fertilization outcome in a total of 233 cycles. It was concluded that the HZA is a good predictor of fertilization rate in vitro, and can be used in the IVF setting to supply additional clinical information in male-factor patients.

Female↗
PubMed1985

Mouse sperm antigens that participate in fertilization. III. Passive immunization with a single monoclonal antisperm antibody inhibits pregnancy and fertilization in vivo.

Passive immunization was used to study the effect of antimouse sperm monoclonal antibodies on fertilization in vivo. The effects of two antibodies were compared in this investigation. One of them, M29, has been shown previously to localize to the equatorial segment of the sperm head and to inhibit mouse fertilization in vitro in a concentration-dependent manner. The second antibody, M2, binds to the same area of the sperm head, and also belongs to the M immunoglobulin class (IgM), but does not affect fertilization in vitro. Superovulated female mice received two antibody injections intraperitoneally (at the times of the pregnant mare's serum gonadotropin and human chorionic gonadotropin injections) at concentrations of 0.5-4.0 mg of IgM or control IgG; animals were mated within 6-12 h of the hCG injection. Fertilization and concomitant establishment of pregnancy were reduced significantly, in a dose-dependent manner, only in those animals immunized with M29 IgM (e.g., 4 mg M29 IgM: 12.6% of 304 eggs fertilized; 4 mg M2 IgM: 96% of 192 eggs fertilized). Intraperitoneal administration of the antibodies did not depress superovulation levels nor oocyte viability. 125I-labeled M29 IgM was used to determine the amount of antibody present in the oviductal ampulla at the time of fertilization in passively immunized mice. Luminal M29 IgM was found to be a linear function of the intraperitoneal dose: 0.002-0.003% of the injected dose was present in the oviductal lumen 14-16 h post-hCG.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Correlation of computerized semen analysis with successful fertilization of oocytes in an in vitro fertilization program.

Sixty-nine couples enrolled in 123 in vitro fertilization-embryo transfer cycles were categorized by percentage fertilization; the results of categorization were compared with those of computerized semen analysis carried out with the CellSoft semen analyzer. Four groups were established: group 1 had 75% fertilization or greater; group 2 had 34% to 74% fertilization; group 3 had 1% to 33% fertilization; and group 4 had 0% fertilization. Statistical differences in certain semen parameters (motility, linearity, and straight-line velocity) were found comparing groups 1 and 3 using the initial ejaculate. A significant number of patients in group 1 had all normal semen parameters, but no statistical difference could be found in group 3 or 4 because of variations in specific abnormal parameters in the groups. When the straight-line velocity-motile density (SLVMD) calculation was used, a significant difference was seen between group 1 and group 3 and between group 1 and group 4 (p less than 0.01); 65% of group 3 and 76% of group 4 had an abnormal SLVMD. SLVMD is a useful calculation to predict fertilization rates in vitro from the initial ejaculate.

Computers↗
PubMed1979

Infertility in the turkey. III. Effects of sera from sterile and fertile females on the fertilizing ability of spermatozoa.

In an attempt to understand the cause for spontaneous infertility in female turkeys, sera were obtained from a flock suffering this condition. These sera, from hens of known fertility, were tested for anti-fertility actions by incubating semen in them. After incubation, semen was inseminated into unrelated recipient hens. In the first of two experiments, semen incubated in sera from high fertility hens (average 89%) produced a fertility level of 72.7% over an 8 week period while semen incubated in sera from sterile hens significantly reduced fertility to 52.3%. In a second experiment sera from sterile donors again caused a significant reduction in the fertilizing ability of sperm (56.3% for sera from sterile hens vs. 73.5% for sera from high fertility hens). These results indicate the presence of a factor(s) in the sera of sterile hens which may be causally related to their infertility.

Animals↗
PubMed2004

[Effects of long-term fertilization on soil microorganism and its role in adjusting and controlling soil fertility].

To clarify the relationships between soil microorganisms and soil fertility under the condition of long-term fertilization, a 12-year fertilization experiment was installed on a fluvo-aquic soil, and the amount of soil microorganisms and the content of soil nutrients were determined and analyzed. The results showed that chemical fertilizers combined with organic manure could significantly improve soil fertility and increase the amount of soil microorganisms. Organic manure was obviously superior to corn straw in improving soil fertility. The correlation between soil microorganisms and soil fertility was significant. A positive correlation was found between the amount of bacteria and azotobacteria and the contents of organic matter, total N, alkalified N, total P and available P. The relationship between the amount of fungi and actinomycetes and the content of soil nutrients was not obvious.

Actinobacteria↗
PubMed2006

[Changes of crop yield and soil fertility under long-term fertilization and nutrients-recycling and reutilization on a black soil: IV. Soil organic carbon and its fractions].

A long-term experiment was conducted on a black soil of Northeast China to study the effects of applying chemical fertilizers and recycled organic manure (ROM) on the changes of soil organic carbon and its fractions. The results showed that from 1985 to 2004, soil total organic carbon (TOC) decreased by 7.83% in control,4.56% in N application, 1.61% in N + P application, and 5.56% in ROM application, but increased by 0.33% in N + P + K application. Comparing with single application of ROM, its application with chemical fertilizers, i. e., N + ROM, N + P + ROM, and N + P + K + ROM, increased the TOC concentration by 0.35%, 1.05% and 0.64%, respectively. The readily oxidized carbon (ROC) in fertilization treatments was increased by 8.64% to approximately 28.4%, and the increment was higher in treatments of chemical fertilizers plus ROM than in treatments of chemical fertilizers. The ROC was significantly correlated with soil TOC (Y = 14.192X + 23.9, R2 = 0.802) and stalk yields (Y = 19032X - 7950.6, R2 = 0.759). Light fraction organic carbon (LF-C) had the same trends with ROC. After 20 years fertilization, the organic carbon in soil humic acid and fulvic acid was decreased by 1.64% to approximately 26.23% and 2.33% to approximately 28.68%, respectively, but in treatments of chemical fertilizers plus ROM, the decreasing trend was slowed down.

Biomass↗
PubMed1990

Fertility desires and fertility: hers, his, and theirs.

The relationship between desired and achieved fertility may be misspecified by excluding husbands' fertility desires or by confounding effects of shared desires with the resolution of conflicting desires. Using couple data from the classic Princeton Fertility Surveys, we find relatively large husband effects on fertility outcomes as well as unique effects of spousal disagreement. Wives and husbands were equally likely to achieve fertility desires, and disagreeing couples experienced fertility rates midway between couples who wanted the same smaller or larger number of children. These conditions do not hold, however, when we include willingness to delay births for economic mobility as part of the measure of fertility desires. Among couples who both wanted a third child, only husbands' willingness to delay births had significant negative effects on birth rates.

Birth Rate↗
PubMed1994

Recent fertility decline in Dariusleut Hutterites: an extension of Eaton and Mayer's Hutterite fertility study.

A church book that has been routinely updated by the Dariusleut Hutterites enabled us to update some fertility tables presented by Eaton and Mayer in the 1950s. The age-specific (nuptial) fertility rates and the total fertility rates (TFRs) were calculated for every 5-year period from 1901-1905 to 1981-1985. Our calculations for Dariusleut, one of the three sects of the Hutterites, gave slightly lower age-specific nuptial fertility rates before 1951 compared with the figures given by Eaton and Mayer (1953) for all Hutterites in the corresponding time periods. The recent decline in Hutterite fertility, especially at higher maternal ages, was confirmed in this study. The TFR, given as a sum of 5-year-grouped age-specific nuptial fertility rates from 15 to 49 years of maternal age, was higher (8.80-9.83) in the years 1946-1965, but it declined to 8.13 in 1966-1970, to 7.22 in 1971-1975, to 6.39 in 1976-1980, and to 6.29 in 1981-1985. For the age groups 35-39 years and older the age-specific nuptial fertility rate in 1981-1985 decreased by more than 50% compared with the rate during the peak period of 1951-1955. The probability of a married woman having a live birth at each age from 15 to 49 years still peaked around the twentieth year of life, as found by Eaton and Mayer (1953), but it decreased more rapidly from the late twenties among the recently married women, most evidently in their thirties.

Adolescent↗