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Further development of a transcervical technique for artificial insemination in sheep using previously frozen semen.

A transcervical technique (the Guelph System for transcervical AI) was used to inseminate 2060 ewes on 65 farms (average 31 ewes, range 5 to 107) in Ontario, Canada, from October 1990 to September 1992, using previously frozen semen. Estrus was synchronized using progestagen pessaries and PMSG with median inseminations done at 54 h from pessary removal. Maiden ewes were not included. Only ewes in which the cervix could be penetrated were inseminated with 150 million spermatozoa per insemination. A total of 1809 were penetrated and inseminated (penetration rate 87.8%). Success of penetration increased from 76.3% in the first 500 ewes to 97.9% in the last 500 (P=0.01). Cervical penetration was more successful in ewes in the accelerated lambing program (92.3%, average 3.1 mo since the previous lambing) than those in the annual lambing program (82.4%, average 7.0 mo since the previous lambing; P=0.06). The lambing rate for ewes bred during the combined traditional breeding seasons (Fall of 1990, 1991, 1992) was 50.7% compared to 24.4% for ewes bred at other periods (P=0.00001). The average time required for handling and insemination decreased from 8.62 min in the first 500 ewes to 3.62 min in the last 500 ewes. The Guelph System for Transcervical AI was found to be successful for cervical penetration in most ewes. Penetration success was affected by period since the last lambing and by inseminator experience. The lambing rate was higher for ewes bred during the traditional Fall breeding seasons than during other times of the year.

Journal Article↗

GnRH-agonist induction of fertile estrus with either natural mating or artificial insemination, followed by birth of pups in gray wolves (Canis lupus).

Although captive populations of endangered species such as the Mexican gray wolf (Canis lupus baileyi) can benefit from artificial insemination to accomplish genetic exchange, reliable techniques for timing insemination are lacking. We used the generic gray wolf (C. lupus) to test the efficacy of a short-acting GnRH-agonist implant, deslorelin, for inducing estrus. Of five females receiving implants on 17 or 18 January 2003, two mated naturally 10-17 days later, and the others were artificially inseminated using fresh semen, one on day 7 and all three on day 11. Relaxin tests revealed that one artificially inseminated female and both naturally mated females were pregnant on 1 March, and all three gave birth to healthy puppies on 4-6 April. Of the artificially inseminated females, only the one who subsequently conceived and gave birth was judged to be in cytologic estrus at the time of insemination. Two females were treated again with deslorelin on 12 January 2004, followed by collection of fecal samples for hormone analysis. One female, who was housed with a male, copulated on day 17 but did not conceive; the other was not with an adult male. Fecal progestin and estrogen profiles suggested that estrus, but not ovulation, was induced. These results indicated that deslorelin could induce fertile estrus in the gray wolf, although individual response varied. Further investigation is needed to better define and control the interval between implant insertion and ovulation for optimal timing of insemination.

Animals↗

The effect of a progesterone releasing intravaginal device (PRID) on pregnancy risk to fixed-time insemination following diagnosis of non-pregnancy in dairy cows.

The objective was to compare the probability of pregnancy after fixed-time insemination in cows diagnosed as non-pregnant and re-inseminated following the Ovsynch protocol, with or without exogenous progesterone. Cows (n=415) used in this study originated from 25 farms. Upon diagnosis of non-pregnancy between 30 and 60 days after AI, cows were randomly assigned to receive either a progesterone releasing intravaginal device (PRID; n=208) or a placebo intravaginal device (PID; n=207). All cows received GnRH at enrollment (Day 0), PGF(2alpha) concurrent with intravaginal device removal 7 days later, GnRH on Day 9 and fixed-time insemination 16h later (Day 10). Cows observed in estrus prior to Day 7, had the device removed and were inseminated. Ovaries were examined by transrectal palpation at the time of enrollment and the prominent structures were assessed and recorded. Body condition score, lameness status, interval from previous insemination, and times bred at enrollment were recorded. At intravaginal device removal, the occurrence and intensity of vaginitis was determined according to the amount of debris on the device. Overall, the intravaginal device retention rate was 91%. A total of 5.2% of PID-treated cows and 2.9% of PRID-treated cows were detected in estrus within the 7 days treatment period. Pregnancy status was diagnosed between 30 and 56 days after insemination and all cows were followed for a minimum of 150 days after enrollment. Approximately 28% of cows had evidence of mild vaginitis in response to the intravaginal device, whereas 6% of cows had copious debris associated with the intravaginal device at removal. The probability of pregnancy after fixed-time insemination was 43.8% versus 34.9% in PRID-treated versus PID-treated animals. Exogenous progesterone provided through an intravaginal device to non-pregnant cows that had not displayed estrus improved the probability of pregnancy after fixed-time AI.

Administration, Intravaginal↗

Insemination of heifers with sexed sperm.

Data from inseminating 1,000 heifers consecutively with sexed sperm and 370 heifers with control sperm in 11 small field trials are summarized. Semen was from 22 bulls of unknown fertility of various beef and dairy breeds, and 6 inseminators participated. Freshly collected sperm were sexed using a MoFlo flow cytometer/cell sorter after staining sperm with the DNA-binding dye Hoechst 33342; the principle is that the bovine X chromosome has 3.8% more DNA than the Y chromosome. Accuracy approaching 90% males or females was achieved. There was little difference in pregnancy rates between sexed, unfrozen and sexed, frozen sperm. In 5 of 6 field trials, there was little difference in pregnancy rates between insemination doses of 1.0 to 1.5 x 10(6) versus 3.0 x 10(6) sexed, frozen sperm. In the most recent trials, pregnancy rates with sexed, frozen sperm were within 90% of unsexed, frozen controls that had 7 to 20 times more sperm/insemination dose; however, in a few trials, control pregnancy rates were substantially higher than with low doses of sexed sperm. There were too few inseminations per bull to test bull differences in pregnancy rates rigorously. Insemination of sexed, frozen sperm bilaterally into the uterine horns produced pregnancy rates similar to insemination into the uterine body in 4 of 5 field trials. Pregnancy rates among inseminators did not differ significantly. There was no excess embryonic death between 1 and 2 months of gestation with pregnancies from sexed sperm, and very few abortions occurred between 2 months of gestation and term. Although rigorous epidemiological studies remain to be done, calves resulting from sexed sperm appear to exhibit no more abnormalities than controls.

Animals↗

Pre- and post-insemination episodes of sexual selection in the fowl, Gallus g. domesticus.

Although much attention has been recently directed to sexual selection arising after insemination from sexual promiscuity, little is known about the mechanisms determining reproductive success after insemination, and the way these mechanisms interact with each other and with selective mechanisms occurring before insemination: mate choice and mate acquisition. Here, we briefly review the findings of an on-going study investigating the mechanisms generating variation in reproductive success at both a pre- and a post-insemination stage in the domestic fowl. Female preference consistently favours socially dominant males before and after insemination. However, although social status mediates the number of sperm that a male inseminates into a female, dominant males may inseminate sperm of lower fertilising quality than their subordinates. We argue that mitochondrial genes may contribute to determine sperm quality, and speculate that the maternal control of mitochondrial genes may prevent sexual selection from operating on males, thus explaining both the lack of a positive correlation between social dominance and sperm quality and the maintenance of variation in male quality in the fowl.

Animals↗

Pregnancy rates with fresh versus computer-controlled cryopreserved semen for artificial insemination by donor in a private practice setting.

From late 1970 through December 1987, 580 women began 733 treatment programs (a total of 3405 insemination cycles) with fresh semen for artificial insemination by donor. The unadjusted pregnancy rate was 48.5% and the cumulative pregnancy rate after seven cycles by life-table analysis was 59.6% with a monthly fecundability of 11.9%. From 1988 to the present, 113 women have begun 115 treatment programs (a total of 371 insemination cycles) with cryopreserved semen for artificial insemination by donor. The unadjusted pregnancy rate to date is 18.3% and the cumulative pregnancy rate after seven cycles by life-table analysis is 48.6% with a monthly fecundability of 5.9%. This report shows that using cryopreserved semen for artificial insemination by donor will take somewhat longer to produce a pregnancy but the ultimate pregnancy rate will be similar to that with fresh semen for artificial insemination by donor.

Adult↗

Intrauterine insemination: the University of Minnesota experience.

Forty-five patients initiated intrauterine insemination between October 1981 and August 1983. Indications for insemination included poor semen (count less than 20 X 10(6)/ml and/or motility less than 40%), poor cervical mucus, presence of sperm antibodies, unexplained poor postcoital tests, or various combinations of the above. During this time period, 374 inseminations were performed in 163 cycles and resulted in eight pregnancies in the 45 patients receiving artificial insemination by homologous donor, for an overall pregnancy rate of 17.4%. The fact that five of the pregnancies occurred in the first insemination cycle and two in the second cycle was felt to indicate a cause-and-effect relationship. A trial of intrauterine insemination in selected patients would appear to be warranted.

Adult↗

Comparison of fecundability with fresh and frozen semen in therapeutic donor insemination.

A series of 401 therapeutic donor insemination (TDI) cycles was analyzed for determining the effect on fecundability of fresh versus cryopreserved semen, the number of inseminations per cycle, recipient age, and the reproductive health of the TDI recipient. We followed a protocol in which inseminations were performed during the periovulatory period determined by urinary luteinizing hormone surge or ultrasound-timed human chorionic gonadotropin injection. The minimum standard for insemination with fresh or cryopreserved semen was a total of 40 X 10(6) grade 3 sperm. We found no decreased fecundability when using frozen semen, rather than freshly ejaculated specimens. The number of inseminations per cycle did not affect the pregnancy outcome after TDI. The reproductive health of the recipient had a significant effect on the pregnancy rate. Fecundability was 15.6% for healthy women, as compared with 6.7% for women with a diagnosis of moderate, severe, or extensive endometriosis, pelvic adhesions, tubal disease, or ovulatory dysfunction. To avoid transmission of human T-lymphotropic virus type III from sperm donor to TDI recipient, we have recently utilized an exclusively frozen TDI program. With well-timed inseminations of 40 X 10(6) grade 3 motile sperm, we have been able to attain pregnancy rates similar to those previously obtained with fresh samples.

Female↗

Donor insemination--a look to the future.

In this article we have reviewed many aspects of donor insemination. The deficiencies in the screening of semen donors especially for sexually-transmitted diseases has been discussed and importance of a rational protocol for recruitment and screening of potential donors has been emphasized. Factors influencing the success and outcome of donor insemination have been reviewed with particular emphasis on providing prognostic guidelines to potential donor insemination couples as to the success and outcome of the treatment. The importance of accurate timing of inseminations in determining the success of donor insemination has also been discussed. However, as has been highlighted throughout this review, there is a lack of specific prospective controlled trials, which are clearly necessary to answer specific important questions. Also emphasized has been the need of these trials to answer such basic questions as the influence of accurate timing on conception rates in specific groups of insemination recipients.

Female↗

Relative efficiency of therapeutic donor insemination using a luteinizing hormone monitor.

A prospective randomized study was performed to evaluate the use of a urinary luteinizing hormone (LH) detection kit with 1 insemination as compared with 2 alternate day inseminations with timing based on previous cycle length and basal body temperature changes. The study involved 60 patients who underwent a total of 264 therapeutic donor insemination cycles using cryopreserved semen specimens. Patients alternated LH-kit timed cycles with cycles timed by non-LH methods for a total of 6 cycles or until pregnancy was achieved. Fecundability rates were 12.3% for LH-kit cycles and 5.3% for non-LH method cycles. The difference in outcome was not statistically significant. However, when the LH kit plus 1 insemination was compared with 2 inseminations timed by conventional methods, there appeared to be a distinct monetary and time expenditure advantage. These findings suggest that sufficient advantage may be derived from use of an LH kit to recommend its use on a routine basis for the timing of therapeutic donor insemination.

Adult↗

Results of artificial insemination at home by the partner with cryopreserved donor semen: a randomized study.

The use of cryopreserved semen offers the possibility of home insemination by the instructed partner. A comparative study was designed whereby participants were randomly allocated to use home or clinic insemination for six cycles. If no pregnancy had occurred after six cycles, the site of insemination was switched to the opposite location for a maximum of six further cycles. Fifty-three women with primary infertility fulfilling all entry criteria entered the study. In the first 6 cycles out of 29 home starters, 13 pregnancies were conceived, whereas in 24 clinic starters 11 pregnancies occurred, yielding no statistical difference in pregnancy rate. Of 138 couples who did not meet the criteria in the same period, 45 opted for home insemination, resulting in 20 home-inseminated pregnancies. Again, for comparable subgroups no statistical difference in pregnancy rate between home and clinic insemination was found.

Evaluation Studies as Topic↗

[Intra-uterine insemination with activated sperm. Results of conception compared in the various types of infertility in spouses].

A total of 180 infertile couples were treated with intra-uterine insemination of homologous capacitated sperm. The fertility disorder was: a cervical factor in 80 couples (44.4%), a male factor in 68 others (37.8%) and unexplained infertility in 32 couples (17.8%). All the patients received ovarian stimulation with HMG, and with ultrasonographic monitoring. Insemination was performed when the diameter of the dominant follicle reached 18 mm, and HCG 5,000 UI was given on the same day; another insemination was performed 40 hours later if the dominant follicle was still present. The 0.5 ml insemination sample was prepared through a migration-capacitation procedure into Earles medium which yielded 15% of the total motile spermatozoa in the ejaculate. A total of 22 pregnancies were recorded (12% of the couples) in 659 therapeutic cycles; the results however differed according to the infertility disorder: 18.8% of pregnancies in the cervical factor group, 15.6% in the unexplained infertility group and 2.9% only in the male factor group. The lowest number of inseminated motile spermatozoa for pregnancy to occur was 0.4 million. A mild hyperstimulation syndrome was noted in 28 cycles (3.1%); no complications of infectious or immunologic origin occurred. It appears that a procedure which results in a satisfactory pregnancy rate when the sperm is normal (cervical or unexplained infertility) yields poor conceptional results when the semen is abnormal. In cases where the male factor is predominant, intra-uterine insemination should not be performed in the periovulatory period but at ovulation time, and therefore requires either an accurate detection of the LH peak or complete hormonal and sonographic assessment in a stimulated cycle.

Chorionic Gonadotropin↗

Intra-uterine insemination versus timed intercourse for cervical hostility in subfertile couples.

BACKGROUND: The postcoital test has poor diagnostic and prognostic characteristics. Nevertheless, some physicians believe it can identify scanty or abnormal mucus that might impair fertility. One way to avoid 'hostile' cervical mucus is intrauterine insemination. With this technique, the physician injects sperm directly into the uterine cavity through a small catheter passed through the cervix; the theory is to bypass the "hostile" cervical mucus. Although most gynaecological societies do not endorse use of intrauterine insemination for hostile cervical mucus, some physicians consider it an effective treatment for women with infertility thought due to cervical mucus problems. OBJECTIVES: The aim of this review was to determine the effectiveness of intrauterine insemination with or without ovarian stimulation in women with cervical hostility who failed to conceive. SEARCH STRATEGY: We searched Cochrane Central Register of Controlled Trials (CENTRAL) on The Cochrane Library Issue 2, 2005, MEDLINE (1966 to June 2005), EMBASE (1980 to June 2005), POPLINE (to June 2005) and LILACS (to June 2005). In addition, we contacted experts and searched the reference list of relevant articles and book chapters. SELECTION CRITERIA: We included randomized and quasi-randomized controlled trials comparing intrauterine insemination with intercourse timed at the presumed fertile period. Participants were women with cervical hostility who failed to conceive for at least one year. DATA COLLECTION AND ANALYSIS: We assessed the titles and abstracts of 386 publications and two reviewers independently abstracted data on methods and results from five studies identified for inclusion. The main outcome is pregnancy rate per couple. MAIN RESULTS: We did not pool the outcomes of the included five studies in a meta-analysis due to the methodological quality of the trials and variations in the patient characteristics and interventions. Narrative summaries of the outcomes are provided. Each study was too small for a clinically relevant conclusion. None of the studies provided information on important outcomes such as spontaneous abortion, multiple pregnancies, and ovarian hyperstimulation syndrome. AUTHORS' CONCLUSIONS: There is no evidence from the published studies that intrauterine insemination is an effective treatment for cervical hostility. Given the poor diagnostic and prognostic properties of the postcoital test and the observation that the test has no benefit on pregnancy rates, intrauterine insemination (with or without ovarian stimulation) is unlikely to be a useful treatment for putative problems identified by postcoital testing.

Biomarkers↗

Cytogenetic analysis of human oocytes after subzonal insemination.

Subzonal insemination has been proposed to achieve fertilization in cases where standard in vitro fertilization has failed. We present the results of chromosome analysis of oocytes after subzonal insemination. Our data suggest that the main cause (76 per cent) of the absence of cleavage after subzonal insemination is the total absence of sperm nucleus evolution of the injected spermatozoa. Our results also suggest that spermatozoa chromatin development is normal after subzonal insemination. Aneuploidy does not seem to be increased in zygotes after subzonal insemination. However, polyploidy was often more important than predicted by the observation of pronuclei (PN). Pronucleus development might be asynchronous and can appear earlier or later than after standard IVF. The cytogenetic risk after subzonal insemination might therefore be triploidy (if a triploid egg is transferred, because only 2 PN were seen) rather than aneuploidy or structural abnormalities.

Cell Nucleus↗

Secrecy: an unresolved issue in the practice of donor insemination.

There is a paucity of information on the well-being of the families created through artificial insemination with donor sperm despite its widespread availability for more than a generation. Physicians who practice donor insemination have historically supported or encouraged secrecy while observing that couples are satisfied and well adjusted after the birth of their donor insemination offspring. However, from the perspective of the child's best interests, it has been argued that accurate genealogic knowledge should be a "right" and that an ethical conflict exists if the practitioner and the couple act together to deceive the child about his or her true genetic identity. The recognition that secrecy raises difficult social, psychologic, and ethical questions while holding considerable practical and emotional appeal for the couple is a major unresolved issue in the practice of donor insemination. It is suggested that further study and evaluation of donor insemination families is needed if accurate counseling and advice are to be provided to couples considering the option of donor insemination.

Advisory Committees↗

Effect of age on intrauterine insemination with frozen donor sperm.

OBJECTIVE: To assess the effect of variables that influence the pregnancy outcome of intrauterine insemination with frozen donor sperm. METHODS: A retrospective analysis of 408 cycles of frozen donor sperm inseminations were studied. Cycle fecundity and cumulative probability of pregnancy were compared according to several variables. RESULTS: The pregnancy rate was 13.5% per treatment cycle and 57.3% per patient. By life-table analysis, the cumulative probability of pregnancy was 75.6% after 12 cycle attempts, but there was a plateau after seven cycles. The cycle fecundity after the seventh cycle was 0.05, compared with 0.16 during the first seven cycles. Age had a profound impact on cycle fecundity. The cycle fecundity for women age 35 or less, 35-40, and over 40 years old were 0.2, 0.12, and 0.06, respectively. The cumulative probability of pregnancy after seven cycles for women 35 years or younger was 88%, compared with 65 and 42% in women 35-40 and over 40 years old, respectively. The number of motile sperm inseminated for pregnant cycles was higher than that for nonpregnant cycles. The fecundity rates for cycles with five or fewer, five to ten, greater than ten to 20, and over 20 million motile sperm inseminated were 5, 11, 16, and 20%, respectively. CONCLUSION: The most significant predictors of the fertility of intrauterine insemination with frozen donor sperm were the women's age and the total number of motile sperm inseminated. Fecundity dropped after seven cycles of treatment.

Adult↗

Leupeptin, a protease inhibitor, blocks insemination-induced flight muscle histolysis in the fire ant Solenopsis.

The effect of the protease inhibitor leupeptin on flight muscle histolysis in queen fire ants was studied by electron microscopy. In untreated animals artificially inseminated, muscle involution was apparent at 6 hr post-insemination and complete by 24 hr post-insemination. However, in animals pre-treated with leupeptin and subsequently artificially inseminated, no morphologic evidence of flight muscle breakdown was seen at any interval between 6 and 24 hr post-insemination. Such information appears to indicate that one or more proteases are involved in the process of insemination-induced muscle atrophy in fire ants. The most likely candidate is a soluble, calcium-activated myofilament-associated protease.

Animals↗

Effect of artificial insemination on submission rates of lactating dairy cows synchronised and resynchronised with intravaginal progesterone releasing devices and oestradiol benzoate.

This study investigated the hypothesis that a reduction in submission rates at a resynchronised oestrus is not due to the resynchrony treatment involving intravaginal progesterone releasing devices (IVDs) and oestradiol benzoate (ODB) but is associated with artificial insemination (AI) at the first synchronised oestrus. In Experiment 1, cows were synchronised for first oestrus with IVDs, with ODB administered at the time of device insertion (Day 0, 2 mg IM) and 24 h after removal (Day 9, 1 mg IM) and PGF(2alpha) injected at the time of device removal. Cows were then either inseminated (I) for 4 days or not inseminated (NI) following detection of oestrus (first round of AI). Every animal was resynchronised for a second round of AI by reinsertion of IVDs on Day 23 with administration of ODB (1 mg IM) at the time of insertion as well as 24 h after removal (Day 32). Cows detected in oestrus and inseminated for 4 days at the second round of AI were resynchronised for a third round by repeating the resynchrony treatment starting on Day 46 and inseminating cows on detection of oestrus for 4 days. In Experiment 2 the same oestrous synchronisation and resynchronisation treatments were used, but the timing of treatments differed. The cows had their cycles either presynchronised (treatment start Day -23) without AI and then resynchronised, starting on Day 0, for the first round of AI for AI at detected oestrus for 4 days, or they were synchronised (treatment start Day 0) for the first round of AI. In Experiment 1, 91.4% (64/70) and 92.6% (63/68) (P = 0.79) of cows in the I and NI treatments, respectively, were detected in oestrus after the initial synchronisation. At the second round of AI, submission rates for insemination were lower in the I group compared to the NI cows (74.5%, 35/47 versus 92.6%, 63/68, respectively; P = 0.007). Pregnancy rates (proportion treated that were classified as becoming pregnant) in I and NI cows 4 weeks (61.4%, 43/70 versus 63.2%, 43/68) and 7 weeks (77.1%, 54/70 versus 69.1%, 47/68) after the AI start date (AISD) did not differ significantly between treatments. In Experiment 2, presynchronisation and then resynchronisation of oestrous cycles before the first round of AI did not affect oestrous detection rates at the first round of AI (100%, 44/44 versus 98.0%, 50/51; P = 0.54), or pregnancy rates 1 week (63.6%, 28/44 versus 60.8%, 31/51; P = 0.70), 4 weeks (72.7%, 32/44 versus 76.5%, 39/51; P = 0.76) and 7 weeks (81.8%, 36/44 versus 88.2%, 45/51; P = 0.40) after AISD compared to cows that had their cycles synchronised for the first round of AI. These findings support our hypothesis that a reduction in submission rates at a resynchronised oestrus is associated with AI at the first synchronised oestrus and not due to a resynchrony treatment involving IVDs and ODB. This study supports the concept that early embryonic loss following AI at a synchronised oestrus could cause a reduction in submission rates following resynchronisation of oestrus, although investigation of the effect of passing an AI catheter or semen components were not studied per se.

Animals↗