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PubMed1997

Effect of the time of artificial insemination with frozen-thawed or fresh semen on embryo viability and early pregnancy rate in gilts.

The aim of the present study was to evaluate the effect of artificial insemination time (before or after ovulation) using either fresh or frozen-thawed boar semen on embryo viability and early pregnancy rate. Seventy-seven prepubertal crossbred (Landrace x Large White x Duroc) gilts were inseminated in 4 treatments. Artificial inseminations were performed 6 h either after (A) or before (B) ovulation using frozenthawed (A-frozen, n = 19; B-frozen, n = 19) or fresh semen (A-fresh, n = 21; B-fresh, n = 18). The gilts were induced to puberty by administration of 400 IU of eCG and 200 IU hCG (sc) followed by 500 IU of hCG (sc) 72 h later. Ovulation was predicted to occur 42 h after the second injection. All animals were slaughtered 96 h after AI. Embryos were collected and classified as viable (5- to 8-cells, morulae, compacted morulae and early blastocysts) and nonviable (fragmented, degenerated and 1- to 4-cell embryos). The total embryo viability rate was: 64.3% (A-frozen), 54.2% (A-fresh), 76.0% (B-frozen), 91.9% (B-fresh); (A-fresh vs B-fresh, P = 0.018; A-frozen vs B-frozen, P = 0.094). It was observed that AI before ovulation resulted in a higher percentage of total viable embryos than AI after ovulation (P = 0.041). The early pregnancy rate, defined as presence of at least one viable embryo, was 78.9, 80.9, 84.2 and 94.4% for A-frozen, A-fresh, B-frozen, B-fresh, respectively. There was no significant difference in the early pregnancy rate among groups. In conclusion, there was a detrimental effect upon total embryo viability rate when AI was performed after ovulation with either frozen-thawed or fresh semen. The total embryo viability rate and the early pregancy rate were not affected by AI with either frozen-thawed or fresh semen regardless of the time of AI.

Journal Article↗
PubMed1986

Immune dysfunction in rabbits associated with chronic administration of enemas and rectal insemination.

NZ rabbits were treated with various combinations of enemas and intrarectal insemination (1 or 3 ml of semen a week) to investigate the effects of intestinal uptake and immunogenicity of seminal components and of an unrelated antigen, bovine serum albumin (BSA), given simultaneously. For 5 months the treatment was limited to enemas and/or semen, and total immunoglobulins and antisperm and antilymphocyte antibodies were determined. Then, without interruption of the treatments, the animals received two courses of three consecutive daily intrarectal administrations of BSA, and the humoral response was determined 7 days after each course of administration. Only 1 of 18 intrarectally inseminated animals responded with production of antisperm antibodies; none had antilymphocyte antibodies. Total immunoglobulins, however, were significantly increased in animals receiving enemas alone (p less than 0.02) or followed by insemination (p less than 0.05). The humoral response to BSA was significantly (p less than 0.01) enhanced by prior administration of enemas but was moderately reduced by simultaneous administration of semen, in a dose-related fashion.

Animals↗
PubMed1995

Delaying transfer to the third day post-insemination, to select non-arrested embryos, increases development to the fetal heart stage.

The purpose of this study was to determine whether delaying embryo transfer by 24 h, until day 3 post-insemination, allowed improved selection of non-arrested embryos for transfer. We have retrospectively analysed pregnancy rates in a large series of patients who had embryo transfer either on day 2 or on day 3 post-insemination over a 27 month period. From January 1990 to March 1992, 567 patients received embryo transfer on day 2, and 661 patients had transfer on day 3 post-insemination, but these transfers were not contemporary. Pregnancy rates were slightly higher in patients who had embryo transfer on day 3 (37%) than in those patients who had their embryos transferred on day 2 (35%), but this difference was not significant. The implantation rate, as measured by the proportion of embryos developing to the fetal heart stage, was significantly higher following transfer on day 3 (23%) than after transfer on day 2 (19%) (P < 0.05), suggesting that selection of viable embryos is improved on day 3. Furthermore, of the embryos which gave rise to a fetal sac, significantly fewer miscarried before the fetal heart stage (P < 0.05) following transfer on day 3 (6%) than after transfer on day 2 (12%). Delaying transfer until day 3 provides a further 24 h to observe embryo development. During this period 16% of embryos arrested or became developmentally retarded; thus waiting until day 3 allowed these embryos to be identified and avoided for consideration for transfer. Embryo transfer may be safely delayed until day 3, and this may help in selecting embryos most likely to implant and develop after transfer.

Adult↗
PubMed1997

Intrafollicular ovarian pregnancy after ovulation induction/intrauterine insemination: pathophysiological aspects and diagnostic problems.

Ovarian pregnancy is a rare variant of ectopic implantation. A case of an intrafollicular ovarian pregnancy after ovulation induction/intrauterine insemination is presented. The woman had primary infertility of 4 years. Diagnostic laparoscopy revealed endometriosis and adhesions. After adhesiolysis and laser vaporization of endometriotic implants, the patient underwent ovulation induction with artificial insemination by husband/intrauterine insemination; she conceived at her second attempt. The pregnancy proved to be an ovarian intrafollicular one. She was treated by right partial ovariectomy. Three months later she conceived spontaneously with an intrauterine pregnancy which is still ongoing. The diagnostic problems resulting from the coexistence of ovarian hyperstimulation and the intrafollicular development of pregnancy are discussed. A re-evaluation of the criteria for the diagnosis of ovarian pregnancy based on the currently available diagnostic methods is proposed. Moreover, the pathophysiology of ovarian and especially intrafollicular implantation is reviewed.

Adult↗
PubMed1998

The predictive value of the zona-free hamster egg penetration test in relation to in-vitro fertilization at various insemination concentrations.

The aim of the study was to evaluate the predictive value of the zona-free hamster egg penetration test (ZHEPT) for success in in-vitro fertilization (IVF) at various insemination concentrations ranging between 0.1 and >0.6 x 10(6)/ml. The ZHEPT was assessed using sperm samples from 87 couples undergoing IVF treatment. A similar test was simultaneously performed on the same semen sample following ionophore induction of the acrosome reaction (ZHEPTii test). Both the tests were poorly correlated with the fertilization rate of IVF at all the insemination concentrations except at >0.6 x 10(6)/ml, when there was good correlation between the ZHEPTii test and the fertilization rate. Following exclusion of two cases with an oocyte problem, further statistical analysis revealed that both the ZHEPT and ZHEPTii tests were poorly correlated with fertilization rate in IVF in this treatment group. This study suggests that the ZHEPT (with and without ionophore induction of the acrosome reaction) has a poor predictive value for the success of fertilization in IVF treatment at any insemination concentration.

Adult↗
PubMed1998

Sex ratio associated with timing of insemination and length of the follicular phase in planned and unplanned pregnancies during use of natural family planning.

This was a multicentred, prospective study of pregnancies among women using natural family planning. The women maintained natural family planning charts of the conception cycle, recording acts of intercourse and signs of ovulation (cervical mucus changes, including peak day and basal body temperature). Charts were used to assess the most probable day of insemination relative to the day of ovulation and length of the follicular phase of the cycle. The sex ratio (males per 100 females) for 947 singleton births was 101.5, not significantly different from the expected value of 105. The sex ratio did not vary consistently or significantly with the estimated timing of insemination relative to the day of ovulation, with the estimated length of the follicular phase or with the planned or unplanned status of the pregnancy. Although these findings may be affected by imprecision of the data, the study suggests that manipulation of the timing of insemination during the cycle cannot be used to affect the sex of offspring.

Family Planning Services↗
PubMed1999

Comparison between intracytoplasmic sperm injection and in-vitro fertilization (IVF) with high insemination concentration after total fertilization failure in a previous IVF attempt.

The aim of this prospective study was to evaluate whether couples with total fertilization failure in a previous in-vitro fertilization (IVF) attempt should be offered an additional IVF treatment with elevated insemination concentration or should be treated with intracytoplasmic sperm injection (ICSI). In 23 cycles 228 sibling metaphase II (MII) oocytes were randomly divided: 143 and 85 oocytes were utilized for ICSI and IVF respectively. Of the 143 injected (ICSI) oocytes, 90 (62.9%) were normally fertilized (two pronuclei), whereas 21 (14.7%) oocytes were damaged by the ICSI procedure. Of the fertilized oocytes 72 (80%) developed into transferable embryos. No fertilization at all was observed in the 85 sibling MII oocytes which were inseminated (P < 0.001). In all 23 cycles at least one embryo, obtained by ICSI, could be replaced. Eight pregnancies were achieved of which six resulted in the delivery of nine healthy children. In conclusion, for couples with no or almost no fertilization of oocytes in previous IVF attempts, ICSI appeared to be far superior to an additional IVF attempt with further elevated insemination concentrations.

Adult↗
PubMed2002

Time from insemination to first cleavage predicts developmental competence of human preimplantation embryos in vitro.

BACKGROUND: The absence of reliable markers for the identification of viable embryos for transfer at the early cleavage stage is likely to contribute to the generally low implantation rates and high incidence of multiple gestation in IVF treatment. In this study, we investigate the relationship between timing of first cleavage and the incidence of blastocyst formation in vitro. METHODS: Couples (n = 70) with at least one embryo remaining after transfer were included in the analyses. All embryos (n = 579) were examined for early cleavage at 25 h after insemination. Following embryo transfer, the remaining embryos (n = 426) were cultured until day 7 of development, and assessed for blastocyst formation. RESULTS: Eighty-five embryos (14.7%) cleaved to the 2-cell stage within 25 h of insemination; 26 of these were selected for transfer on day 2. Of the 59 embryos remaining in culture, 19 (32.2%) developed to the blastocyst stage; this was a significantly higher number than was observed in embryos (61/367; 16.6%) that failed to cleave within 25 h of insemination (P < 0.01). Within these two groups of embryos the proportion of hatched blastocysts was 11/59 (18.6%) and 26/367 (7.1%) respectively (P < 0.005). CONCLUSIONS: These findings indicate that early cleavage is indicative of increased developmental potential in human embryos and may be useful as an additional criterion in the selection of embryos for transfer.

Adult↗
PubMed2005

Favourable pregnancy results with insemination of in vitro matured oocytes from unstimulated patients.

BACKGROUND: The purpose has been to develop an in vitro oocyte maturation (IVM) method for a wide range of patients. METHODS: A total of 239 cycles with immature oocyte retrieval (IOC) were carried out without hormonal priming. Patients with regular cycles and normal or polycystic ovaries (PCO) and anovulatory cycles with PCOS were included. Insemination or intracytoplasmic sperm injection (ICSI) according to sperm quality was alternatively used in fertilization of the matured oocytes. RESULTS: A total of 971 immature oocytes (mean 8.0 +/- 5.2) were collected in 122 IVM-IVF cycles and 851 oocytes (mean 7.3 +/- 4.4) in 117 IVM-ICSI cycles. The oocyte maturation and fertilization rate was 62.6% and 37.7% after insemination, and 53.9% and 69.3% after ICSI, respectively. The mean number of embryos transferred was 1.6. Clinical pregnancy rate per IOC was 23.8% in IVM-IVF and 17.1% in IVM-ICSI (ns). Implantation rate was higher in IVM-IVF (24.2%) than in IVM-ICSI (14.8%) (P < 0.05). CONCLUSIONS: Insemination of IVM oocytes functions well, resulting in comparable pregnancy rates per IOC between IVM-IVF and IVM-ICSI. Good pregnancy results can be achieved both in patients with regular cycles and with PCO(S) by transferring only one or two embryos at a time.

Cells, Cultured↗
PubMed1995

Timing of pronuclear development and first cleavages in human embryos after subzonal insemination: influence of sperm phenotype.

The sequential transformations of human sperm nuclei in human eggs after subzonal insemination (SUZI; n = 104) and the influence of sperm defects on this timing were studied. This chronology was compared to that of two control series of zygotes obtained after SUZI with normal spermatozoa (n = 35) and after in-vitro fertilization (IVF) with normal donor spermatozoa (D-IVF; n = 220). Pronuclear formation took place between 4.5 and 10.5 h post-SUZI for 92.8% of the zygotes. They remained visible for 13 h and began to disappear 18.5 h post-SUZI. The timespan 3 h. Zygotes obtained after D-IVF had a similar rate of pronuclear disappearance but approximately 4 h later. The second cell cycle was more rapid for zygotes obtained by D-IVF than by SUZI, but the developmental rate of zygotes obtained by SUZI varied according to sperm phenotypes. For patients with previous unexplained IVF failures (control group with normal spermatozoa), the developmental rate was lower, suggesting the influence of oocyte quality. In conclusion, the end of the first cell cycle of zygotes obtained by insemination under the zona pellucida appears 4 h earlier compared to zygotes obtained after insemination outside the zona pellucida.

Adult↗
PubMed1992

Influence of sperm parameters on outcome of subzonal insemination in the case of previous IVF failure. off.

Subzonal insemination (SUZI) has been proposed for patients with sperm male factor infertility, and in cases of in-vitro fertilization (IVF) failures. However despite SUZI, there still remain couples with very low fertilization rates and even with fertilization failures. Since sperm parameters are known to influence the IVF fertilization rate, we investigated the relation between sperm parameters and the SUZI issue in cases of previous IVF failures with normal or subnormal sperm. Twenty-seven couples were included in the study and were split into two groups according to whether they had normal or subnormal sperm. In the first part of the study a randomized prospective trial comparing SUZI to classic IVF insemination was carried out (11 cycles). In the second part, all the oocytes had SUZI (35 cycles). None of the control inseminated oocytes fertilized. Including all the cases, the fertilization rate after SUZI was 29.8% with a pregnancy rate of 15.2% per cycle. We concluded that: (i) SUZI is efficient for achieving fertilization in cases of IVF failures; (ii) the post SUZI fertilization rate is inversely correlated to the percentage of acrosome defects in the semen (P < 0.001); and (iii) when the sperm is normal, the oocyte quality might be responsible for the previous IVF failures. In spite of a good fertilization rate for this indication, it seems that the chance of having a baby is low.

Adult↗
PubMed1982

Separation of bovine spermatozoa by density on water insoluble Newtonian gels and their use for insemination.

Bovine spermatozoa were separated into different density subpopulations utilizing water insoluble hydrocarbon and silicone gels of defined specific gravity. Sperm density profiles were generated for 13 bulls. The separations were found to be repeatable and characteristic of the bull examined. Considerable density variation among animals was demonstrated. Analysis of the separated spermatozoa, before and after freezing, demonstrated that good motility and acrosomal integrity of spermatozoa were maintained. When the least dense fraction of spermatozoa was used for insemination, conception rates were similar to those obtained routinely by artificial insemination with unfractionated spermatozoa. Therefore, this system may be useful in separating spermatozoa of various densities and for removing extraneous matter from semen. However, the sex ratio, among 51 60-day-old fetuses recovered from heifers inseminated with the lowest density fraction of spermatozoa, was 26 males:25 females.

Acrosome↗
PubMed2003

Reducing a cost of traumatic insemination: female bedbugs evolve a unique organ.

The frequent wounding of female bedbugs (Cimex lectularius: Cimicidae) during copulation has been shown to decrease their fitness, but how females have responded to this cost in evolutionary terms is unclear. The evolution of a unique anatomical structure found in female bedbugs, the spermalege, into which the male's intromittent organ passes during traumatic insemination, is a possible counteradaptation to harmful male traits. Several functions have been proposed for this organ, and we test two hypotheses related to its role in sexual conflict. We examine the hypotheses that the spermalege functions to (i) defend against pathogens introduced during traumatic insemination; and (ii) reduce the costs of wound healing during traumatic insemination. Our results support the 'defence against pathogens' hypothesis, suggesting that the evolution of this unique cimicid organ resulted, at least partly, from selection to reduce the costs of mating-associated infection. We found no evidence that the spermalege reduces the costs of wound healing.

Animal Structures↗
PubMedDate not supplied

Influence of the concentration of motile sperm inseminated on the ovum penetration assay results: toward a standardized method.

The influence of the concentration of motile sperm inseminated on the zona-free hamster egg penetration rate has been studied. In 3 fertile men, the higher the concentration of motile sperm inseminated, the higher the ovum penetration rate. This correlation emphasizes the usefulness of standardizing the technical conditions under which the ovum penetration assay is performed. It is also suggested to refer to a standard curve for results analysis. In the 22 patients studied, the outcome of the heterologous ovum assay was in most cases related to the initial sperm parameters. However, some disagreement was also observed. In the absence of a correlation between the initial sperm parameters and the ovum penetration rates, further investigation is needed in order to either more precisely determine the aetiology of unexplained couple infertility, or, in case of intrauterine insemination or in vitro fertilization, to better understand the conditions under which spermatozoa are capacitated in vitro.

Animals↗
PubMed1988

Chromosome analysis of preovulatory human oocytes and oocytes failing to cleave following insemination in vitro.

These investigations were carried out within a clinical in vitro fertilization and egg replacement program. A gradual fixation method for chromosome preparations of animal oocytes, when modified and applied to human oocytes, gave numerical analyzability in about 80%. Of preovulatory oocytes recovered in stimulated cycles, 53% of oocytes with numbers of chromosomes in the haploid range had an apparently normal set of 23 chromosomes. Oocytes with a very low number of chromosomes or in first meiosis were also found. In a separate investigation the chromosome constitution of oocytes not showing clinical signs of fertilization 48 hours after insemination in vitro was also studied. Of the oocytes with numbers of chromosomes in the haploid range, 50% had a normal 23,X karyotype. A different distribution of abnormal chromosome preparations was found, compared with the preovulatory oocytes. Twelve oocytes (16%) carrying between 10 and 18 chromosomes were found among the inseminated oocytes, but none in the other study. Moreover, 15 preparations displayed no chromosomes in the inseminated oocytes, whereas in the uninseminated group, none lacked chromosomes completely. These results may indicate how unfertilized human oocytes in vitro undergo chromosome loss and degeneration and could represent different stages of degeneration. Among both preovulatory and unfertilized oocytes the distribution between hyper- and hypohaploid oocytes was of particular interest in that hypohaploid chromosome complements were in excess. This finding supports the hypothesis that aberration in the number of chromosomes in an oocyte is caused not only by nondisjunction but also by anaphase lag, since nondisjunction alone would result in a 1: 1 ratio between hyper- and hypohaploid complements. In conclusion, human oocytes recovered for in vitro fertilization seem to have an incidence of numerical chromosome abnormalities in the region of 40-50%. This is close to the figures estimated on the basis of observations made in spontaneous abortions and could be a major explanation for the low developmental potential of human cleavage stage ova.

Cell Division↗
PubMed1994

Embryo cleavage rates after oocyte insemination with capacitated sperms in the presence or absence of follicular fluid.

The objective of the present study was to compare the rate of embryo cleavage after oocyte insemination with capacitated spermatozoa in the presence or absence of follicular fluid. In the first experiment, a total of 67 oocytes were inseminated with spermatozoa which had been submitted to part of the capacitation procedures in the presence of follicular fluid (21 oocytes) and with spermatozoa capacitated only with Menezo B2 culture medium (46 oocytes). Thirteen and 29 cleaved embryos were obtained, respectively (p = 0.89). In the second experiment, a total of 61 oocytes were inseminated with spermatozoa submitted to the entire capacitation procedure in follicular fluid (17 oocytes) and capacitated only with Menezo B2 culture medium (44 oocytes). Eleven and 31 cleaved embryos were obtained, respectively (p = 0.67). The absence of significant differences in cleavage rate between the two experiments indicates that the quality of the spermatozoa capacitation process is not changed by the addition of follicular fluid.

Cleavage Stage, Ovum↗
PubMed2002

Intrauterine pregnancy following low-dose gonadotropin ovulation induction and direct intraperitoneal insemination for severe cervical stenosis.

BACKGROUND: We present a case of primary infertility related to extreme cervical stenosis, a subset of cervical factor infertility which accounts for approximately 5% of all clinical infertility referrals. CASE PRESENTATION: A 37 year-old nulligravida was successfully treated with ovulation induction via recombinant follicle stimulating hormone (FSH) and direct intraperitoneal insemination (IPI). Anticipating controlled ovarian hyperstimulation with in vitro fertilization/embryo transfer (IVF), the patient underwent hysteroscopy and cervical recanalization, but safe intrauterine access was not possible due to severe proximal cervical stricture. Hysterosalpingogram established bilateral tubal patency and confirmed an irregular cervical contour. Since the cervical canal could not be traversed, neither standard intrauterine insemination nor transcervical embryo transfer could be offered. Prepared spermatozoa were therefore placed intraperitoneally at both tubal fimbria under real-time transvaginal sonographic guidance using a 17 gage single-lumen IVF needle. Supplementary progesterone was administered as 200 mg/d lozenge (troche) plus 200 mg/d rectal suppository, maintained from the day following IPI to the 8th gestational week. A singleton intrauterine pregnancy was achieved after the second ovulation induction attempt. CONCLUSIONS: In this report, we outline the relevance of cervical factor infertility to reproductive medicine practice. Additionally, our andrology evaluation, ovulation induction approach, spermatozoa preparation, and insemination technique in such cases are described.

Journal Article↗
PubMed2006

In vitro culture conditions using chemically defined media for in vitro matured and intracytoplasmically inseminated porcine oocytes.

The present study investigated in vitro culture methods [droplet and Well of the Well (WOW)] using semi-defined and defined media [modified porcine zygote medium (mPZM)] and the additional effects of insulin on in vitro matured and intracytoplasmically inseminated porcine oocytes. In Experiment 1, in vitro matured and intracytoplasmically inseminated porcine oocytes were cultured for 6 days in the following four groups: 1) mPZM-3 (containing bovine serum albumin) + droplet (30 mul), 2) mPZM-3 + WOW, 3) mPZM-4 (containing polyvinyl alcohol) + droplet, and 4) mPZM-4+ WOW. The culture media (mPZM-3 and mPZM-4) and methods (droplet and WOW) did not significantly affect the cleavage rate, but the blastocyst rate of the oocytes cultured in mPZM-3 was significantly (P<0.01) higher than that of mPZM-4 (20.1 and 9.4%, respectively). The blastocyst rates as percentages of the cleaved oocytes (51.8 and 16.9%) and the hatched blastocyst rate as percentages of the number of blastocysts (12.3 and 2.2%) were also significantly (P<0.01) higher in mPZM-3 compared with those in mPZM-4. There was significant interaction (P<0.05) between the two main factors; the effects of the culture media and methods on the rate of hatched blasyocysts as percentages of the blastocysts produced and, the hatched blastocyst rate (20.3%) as percentages of the number of blastocysts produced in mPZM-3 were significantly (P<0.05) higher than in the other groups. In Experiment 2, the additional effects of insulin (100 ng/ml) in mPZM-3 and mPZM-4 media was investigated in the WOW culture system. Insulin addition did not improve cleavage, blastocyst formation, or the number of cells in blastocysts. However, as in Experiment 1, mPZM-3 resulted in a significantly higher blastocyst rate as percentages of the cleaved oocytes than mPZM-4 (33.9 and 18.4%). These results indicate that a chemically defined medium (mPZM-4) needs to be improved to provide more suitable culture conditions for in vitro development of in vitro matured and intracytoplasmically inseminated porcine oocytes. However, the WOW system may be a useful IVC method for blastocyst development of in vitro matured porcine oocytes following ICSI when a semi-defined medium (mPZM-3) is used.

Animals↗