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Offspring from inseminations with mammalian sperm stained with Hoechst 33342, either with or without flow cytometry.

The technique of vital staining with a fluorochrome and subsequent flow cytometry has been employed to investigate the DNA content of living sperm. There have been several reports of chromosomal damage caused by staining cells with a bis-benzimidazole dye. Hoechst 33342, either with or without subsequent flow cytometry. Therefore the use of this technique might be expected to affect adversely the fertilising capacity of inseminated sperm, or to result in the production of congenital deformities in the young. Insemination experiments to date have resulted in the production of more than 400 offspring in four species, including 5 successive generations of rabbits, where stained or sorted sperm were used. All progeny were observed to be normal by anatomical criteria. There was no evidence to suggest that sperm treated in this manner were unable to fertilise oocytes in vivo or that embryonic development was affected.

Animals↗

Influence of donor insemination on sexual functions of recipients.

This paper presents the results of an investigation by personal interviews by the author, concerning different characteristics of sexual behaviour in 156 women during the period in which donor insemination was performed. The questions concerned sexual desire, orgasm, masturbation, libido alterations during the menstrual cycle, libido of wife compared to libido of husband, influence of sex films and sex books on sexual desire. The conclusion that can be drawn from this study is that donor insemination did not disturb the sexual part of marital life in the interviewed group of women.

Female↗

Factors affecting conception rate after artificial insemination and pregnancy loss in lactating dairy cows.

Objectives were to determine factors associated with conception rate (CR) and pregnancy loss (PL) in high producing lactating Holstein cows. In Study 1, CR was evaluated in 7633 artificial inseminations (AI) of 3161 dairy cows in two dairy farms. Pregnancy diagnosis was performed by palpation per rectum 39+/-3 days after AI. Environmental temperature was recorded at different intervals prior to and after AI. In Study 2, 1465 pregnancies from 1393 cows diagnosed at 31+/-3 days after AI by ultrasonography on three dairy farms were re-examined 14 days later to determine PL. Temperature > or =29 degrees C was considered to be heat stress (HS). Exposure to HS was defined as following: NH, no heat stress; HS1, exposure to at least 1 day of maximum temperature > or =29 degrees C and average daily maximum temperature (ADMT) <29 degrees C; and HS2, exposure to ADMT > or =29 degrees C. In Study 1, exposure of cows to HS1 and HS2 from 50 to 20 prior to AI was associated with reduced CR compared to cows not exposed to HS (28.8, 23.0, and 31.3%, respectively). Post-AI HS was not associated with CR. Cows inseminated following estrus detection or timed AI had similar CR. As the number of AI increased, CR decreased. Multiparous cows had lower CR than primiparous cows, and occurrence of milk fever and retained placenta was associated with decreased CR. In Study 2, PL was not associated with exposure to HS either prior to or after AI. Cows diagnosed with clinical mastitis experienced increased PL, but parity, number of AI, AI protocol, milk production, and days postpartum at AI were not associated with PL. In conclusion, CR was affected by HS prior to AI, parity, number of AI, and postparturient diseases, whereas PL was affected by clinical mastitis.

Abortion, Veterinary↗

Homologous intrauterine insemination in controlled ovarian hyperstimulation cycles: a comparison among three different regimens.

OBJECTIVE: The objective was to assess the efficacy of double intrauterine insemination (IUI) over a single periovulatory IUI in patients undergoing controlled ovarian hyperstimulation with low-dose recombinant follicle stimulating hormone (rFSH) combined with human chorionic gonadotropin (HCG). STUDY DESIGN: Ninety-four infertile women were randomly assigned to three groups; in group A (38 patients, 47 cycles) a single IUI was performed 36 h after HCG administration combined with timed intercourse the day of HCG administration; within group B (43 patients, 48 cycles) IUI alone was performed 36 h after HCG administration; in group C (39 patients, 43 cycles) a double IUI 12 and 36 h after HCG administration was performed. RESULTS: The mean age and the causes of infertility were similar between the three groups. The number of follicles greater than 15 mm on the day of HCG administration and the overall dose of rFSH required per cycle was not significantly different among the groups. The pregnancy rate (PR) per cycle and per patient was 14.9% and 18.4% in group A, 10.4% and 11.6% in group B, 20.9% and 23.1% in group C, respectively. There was no statistically significant difference in PR among the three groups. CONCLUSION: In rFSH/HCG cycles, two IUIs performed 12 and 36 h after HCG administration do not significantly improve pregnancy rates over a single insemination performed 36 h after HCG administration combined with or without timed intercourse the day of HCG administration.

Adult↗

Women with ovulatory dysfunction undergoing ovarian stimulation with clomiphene citrate for intrauterine insemination may benefit from administration of human chorionic gonadotropin.

OBJECTIVE: To investigate factors that may affect pregnancy outcome following ovarian stimulation with clomiphene citrate (CC) combined with intrauterine insemination (IUI). DESIGN: Retrospective cohort study. SETTING: University teaching hospital. PATIENT(S): Three hundred and twenty women who underwent 691 ovarian stimulation cycles with CC for IUI. INTERVENTION(S): Ovarian stimulation with CC followed by a single IUI either 24 hours after a spontaneous serum LH surge (>25 mIU/mL) or 36 hours after intramuscular human chorionic gonadotropin (hCG) administration (10,000 IU) when the largest follicle had reached a diameter of 17 mm. MAIN OUTCOME MEASURE(S): Clinical pregnancies. RESULT(S): Women with ovulatory dysfunction who received hCG had significantly higher pregnancy rates (24.6%) compared with women with other types of infertility. There were no differences in pregnancy rates between the LH surge group and the hCG group (14.3% vs 12.4%). A spontaneous LH surge was noted in a variety of follicular sizes (14 to 35 mm). There was no correlation for age, body mass index, follicular diameter, number of mature follicles, other sperm characteristics, and pregnancy outcome in either group. CONCLUSION(S): After ovarian stimulation with CC, IUI is equally effective 24 hours after a spontaneous LH surge or 36 hours after administration of hCG. Spontaneous LH surges were observed at a variety of follicular sizes with comparable pregnancy rates. In women with ovulatory dysfunction, hCG administration before insemination may be beneficial.

Adult↗

Flexible versus rigid intrauterine insemination catheters: a prospective, randomized, controlled study.

OBJECTIVE: To prospectively compare pregnancy rates for couples undergoing IUI by a single healthcare provider with random assignment to either rigid (Tomcat; Kendall Sovereign, Mansfield, MA) or flexible (Soft Pass; Cook, Spencer, IN) catheters. DESIGN: Randomized, controlled study. SETTING: Tertiary care infertility practice. PATIENT(S): One hundred infertile women enrolled from a single infertility practice who met the inclusion and exclusion criteria for IUI were enrolled. Of the 100 patients, 95 underwent ovarian stimulation before IUI: 35 with clomiphene citrate alone, 60 with clomiphene citrate combined with a single dose of gonadotropins. INTERVENTION(S): Timed IUI was performed with either rigid (Tomcat) or flexible (Cook) insemination catheters. MAIN OUTCOME MEASURE(S): Pregnancy rate per cycle. RESULT(S): No differences were noted between groups for the following: age, length of infertility, day-3 FSH level, number of prior IUI cycles, total motile sperm count, days of abstinence, infertility diagnoses, insemination ratings, and stimulation protocols. No difference in pregnancy rates was observed between rigid and flexible catheter groups. CONCLUSION(S): There is no statistically significant difference between flexible and rigid catheters for IUI.

Adult↗

Effect of ejaculatory abstinence period on the pregnancy rate after intrauterine insemination.

OBJECTIVE: To determine the optimal interval of ejaculatory abstinence for couples undergoing IUI. DESIGN: Retrospective analysis. SETTING: Reproductive endocrinology and infertility center. PATIENT(S): Infertile couples undergoing ovulation induction and IUI with partner's semen. INTERVENTION(S): Ovulation induction with clomiphene citrate and a single IUI procedure per cycle. MAIN OUTCOME MEASURES(S): Clinical pregnancy rates as a function of abstinence intervals. RESULT(S): Four hundred seventeen women underwent 929 cycles from June 1999 to October 2002 for a median of 4 IUI attempts per couple. The median ejaculatory abstinence interval was 4 days (range 0-30) with an overall pregnancy rate of 12% per cycle. Abstinence correlated positively with inseminate sperm count but negatively with motility. Variations in inseminate parameters did not correlate with pregnancy rates. However, abstinence intervals significantly affected pregnancy rates. The highest pregnancy rate was observed with an abstinence interval of 3 days or less (14%) and the lowest pregnancy rate seen with an abstinence interval of 10 days or more (3%). CONCLUSION(S): An abstinence interval of 3 days or less was associated with higher pregnancy rates following IUI. Prolonged abstinence decreases pregnancy rates, independent of other sperm parameters, perhaps as a result of sperm senescence and functional damage not readily identified by standard semen analysis. Abstinence intervals should be controlled for in studies examining pregnancy outcome in assisted reproduction.

Adult↗

Clomiphene citrate monitoring for intrauterine insemination timing: a randomized trial.

OBJECTIVE: To compare pregnancy rates with two different methods of intrauterine insemination (IUI) timing in patients treated with clomiphene citrate (CC). DESIGN: Prospective, randomized trial. SETTING: Academic medical center. PATIENT(S): One hundred fifty ovulatory, infertile women. INTERVENTION(S): Patients were randomized into a luteinizing hormone (LH) surge group or a follicle monitoring/human chorionic gonadotropin (hCG) group. All patients underwent baseline ultrasound, and took clomiphene citrate, during days 5-9. Patients in the LH surge group underwent IUI on the day after a home test for the LH surge was positive, whereas those in the hCG group received hCG according to ultrasound parameters and underwent insemination 33-40 hours later. Patients remained in the same study group for up to three cycles. MAIN OUTCOME MEASURE(S): Pregnancy rates per cycle and per patient. RESULT(S): No significant differences were found between groups in pregnancy rates per patient or per cycle. The LH surge group underwent IUI significantly later than the hCG group. Cancellation rates were significantly higher for the LH surge group (31% vs. 11%) and attributable mainly to failure to detect an LH surge. The majority of pregnancies in both treatment groups occurred in the first cycle. CONCLUSION(S): The decision to use hCG for IUI timing should be influenced by factors other than pregnancy rates.

Adult↗

Preventing high-order multiple pregnancies during controlled ovarian hyperstimulation and intrauterine insemination: 3 years' experience using low-dose recombinant follicle-stimulating hormone and gonadotropin-releasing hormone antagonists.

OBJECTIVE: To employ protocols of mild ovarian stimulation to prevent an excessively elevated rate of high-order multiple pregnancies. DESIGN: Case series. SETTING: University hospital. PATIENT(S): Six hundred and twenty one consecutive patients undergoing 1,259 controlled ovarian hyperstimulation and intrauterine insemination cycles. INTERVENTION(S): Patients received 50 IU per day of recombinant follicle-stimulating hormone (FSH) starting the third day of the cycle, then a gonadotropin-releasing hormone (GnRH) antagonist on the day in which a follicle > or =13 mm was visualized. Cycles were canceled if three or more follicles > or =16 mm and/or five or more follicles > or =11 mm were detected. MAIN OUTCOME MEASURE(S): Rate of high-order multiple pregnancies. RESULT(S): The clinical pregnancy rate per initiated cycle was 9.2% (95% confidence interval, 7.5-11.1%). The incidence of twins and high-order multiple pregnancies was 9.5% (95% CI, 5.3-16.2%) and 0 (0.0-3.2%), respectively. CONCLUSION(S): In controlled ovarian hyperstimulation and intrauterine insemination cycles, a protocol of 50 IU of recombinant FSH daily combined with the use of GnRH antagonists and a policy of strict cancellation based on echographic criteria are associated with a satisfactory pregnancy rate per initiated cycle and a low risk of high-order multiple pregnancies.

Adult↗

[Is there a future for Fallopian tube insemination in women?].

In unselected patients, the pregnancy rate after intrauterine insemination (i.u.i.) seldom goes beyond 10-15% per cycle. An insufficient number of spermatozoa at the fertilization site has been hypothesized for a long time to explain the low efficacy of this technique. Thus, the introduction of a larger number of male gametes into the female tubes has been thought of to give better results since the late eighties. First, a direct tubal catheterisation has been proposed for injection of spermatozoa, either by laparoscopy or transvaginally under ultrasound guidance or by tactile sensation. However, these procedures have been abandoned because of some severe traumatic and infectious complications. Alternatively, a spermatozoa suspension of several millilitres can be injected under pressure into the uterine cavity while sealing the cervical os, by various systems. This technique called Fallopian sperm perfusion (FSP) has yielded some interesting results, particularly in unexplained infertility. Nevertheless, the superiority of FSP over i.u.i. still remains controversial. This review describes the current knowledge about intratubal insemination and its potential role in the management of human infertility.

Adult↗

[Artificial insemination with donor frozen sperm].

The cryopreservation of human sperm decreases its fecondity, by reduction in the number of motile sperm. However the fertilizing ability of the spermatozoa is not faded. The use of frozen sperm requires an optimization of the methods of artificial insemination to hope to approach the results obtained with fresh sperm. Intrauterine insemination with sperm selection meets this aim.

Cryopreservation↗

Semen characteristics, cryopreservation, and successful artificial insemination in the Blue rock pigeon (Columba livia).

The present study was undertaken in the Blue rock pigeon (Columba livia) to evaluate the annual semen characteristics, to identify a suitable extender for semen short-term storage, to determine a protocol for cryopreservation of semen and finally to check whether intracloacal insemination would lead to the birth of a chick. Semen characteristics such as semen volume, sperm concentration, sperm motility, and percentage of normal spermatozoa were maximum during the monsoon season. TALP was observed to be the most suitable semen extender and the sperm survived best at 37 degrees C at a dilution of 1:100 in TALP. Further, cryopreservation studies on pigeon semen indicated that 8% DMSO with or without egg yolk (20%) proved to be a better cryoprotectant compared to glycerol and polyethylene glycol. In addition, the slow freezing protocol was better than the fast-freezing protocol and about 40% of the cryopreserved spermatozoa were motile following thawing. Computer-aided semen analysis indicated that pigeon spermatozoa were extremely active immediately after dilution in TALP and exhibited linear trajectories persisting up to 9h. But, with time there was a time-dependent decrease in the velocity parameters (VAP, VSL, and VCL). Cryopreserved spermatozoa following thawing also exhibited linear trajectories but had reduced velocity as evident from the significant decrease in VAP, VSL, and VCL. Further, artificial inseminations using fresh semen resulted in 45% fertilization and birth of a live chick.

Animals↗

Effect of post-insemination supplementation with PRID on pregnancy in repeat-breeder Holstein cows.

Embryonic mortality contributes to repeat-breeding in dairy cows; luteal insufficiency is a known cause of embryonic mortality. The objective of this study was to assess the efficacy of supplementation with exogenous progesterone for 14 days on pregnancy maintenance in inseminated repeat-breeder dairy cows. On Day 5 after insemination, treated cows ( n=143 ) received a modified PRID (i.e. without estradiol capsule), which was removed on Day 19. Control cows ( n=148 ) did not receive any treatment. Overall there was no effect of PRID supplementation on pregnancy rates. However, when the study population was stratified by parity and stage of lactation, PRID supplementation significantly improved pregnancy rate in first and second parity late lactation cows (risk ratio = 3.26; 95% CI 1.22, 8.69). Pregnancy rates did not differ between PRID-treated cows with ( n=81 ) and without vaginitis. Control cows tended ( P=0.077 ) to have a higher proportion of abortions than PRID-treated cows (7/50 versus 2/51, respectively). In conclusion, young late lactation repeat-breeder cows benefited from progesterone supplementation, in terms of maintaining pregnancy until traditional time of pregnancy diagnosis.

Animals↗

Luteal function and conception in lactating cows and some factors influencing luteal function after first insemination.

The objective of this study was to investigate the types and incidence of luteal sub-function in lactating cows after artificial insemination (AI) and their relationship with pregnancy, and to clarify the relationship between luteal function and parity, body condition score (BCS), milk yield, and dietary intake. In 19 cows, milk samples were collected daily from AI to confirmation of pregnancy. Milk progesterone concentrations were determined by EIA. Based on peak progesterone concentration and the day of onset of luteal phase, 15 of 30 progesterone profiles (50%) were normal, with progesterone concentration reaching 1.0 ng/ml within 5 days after insemination and > or =2.0 ng/ml thereafter. In addition, 6 (20%) were insufficient, (progesterone concentration remained < 2.0 ng/ml), 5 (17%) were delayed (progesterone reached 1.0 ng/ml after 5 days), 2 (7%) were both delayed and insufficient, one (3%) was short (progesterone >1.0 ng/ml for only 7 days), and one (3%) remained basal. Cows with a normal profile had a higher (P < 0.05) pregnancy rate than those with an abnormal profile (87% versus 33%, respectively). The amount of progesterone secreted in milk after first AI, as indicated by progesterone area under curve (AUC), was negatively correlated with milk yield (r = -0.83, P < 0.01), dry matter intake (r = -0.81, P < 0.05), total digestible nutrients (r = -0.82, P< 0.05), and digestible crude protein (r = -0.79, P <0.05). Cows that produced more milk and consumed more dry matter had less progesterone during the luteal phase. In conclusion, abnormal luteal function was associated with reduced pregnancy rates and high milk production and increased dietary intake during breeding were associated with reduced progesterone concentrations.

Animals↗

Effect of the interval between estrus onset and artificial insemination on sex ratio and fertility in cattle: a field study.

We have carried out a field trial in cattle to study the effect of the interval between the onset of estrus and AI on sex ratio and fertility. Data were obtained from 716 cows that had been inseminated at different times between 8 and 44 h from the visual detection of estrus. Before analyzing the data, it was grouped in three intervals considering the time between estrus onset and AI (8-18, 18-30, and > or = 30 h). Our results show that the percentage of calved females (73.05%) is significantly superior for early inseminations (8-18 h), and it decreases 1.85% per hour from the onset of estrus. Delayed AIs (> or = 30 h) produce a significant deviation of the sex ratio towards the males (72.06%); nevertheless, fertility (percentage of successful pregnancies) diminishes significantly, from 66.19% (8-18 h) to 45.35% (> or = 30 h). In conclusion, variations in the interval between the onset of estrus and AI modify sex ratio. However, we must consider its effect on fertility.

Animals↗

Sperm survival and heterogeneity are correlated with fertility after intrauterine insemination in superovulated ewes.

Efficient animal production involves accurate estimations of fertilizing ability. One key factor is the plasma membrane of the sperm cell, which is actively involved in the cascade of events before oocyte fusion. Many methods are used to analyze the characteristics of this membrane, including partition in aqueous two-phase systems which is an efficient method to analyze sperm surface changes accounting for loss of viability and different functional states. Centrifugal countercurrent distribution (CCCD) analysis can also be used in an aqueous two-phase system to determine the relationship between sperm parameters and in vivo fertility in ewes. In a previous work, we found a significant correlation between two post-CCCD parameters (heterogeneity and recovered viability) and field fertility when the same sample was used after cervical AI. The present study was intended to find out whether the control of several external factors that affect reproductive efficiency is able to increase the correlation coefficient between post-CCCD parameters and fertility. Thus, 90 Rasa aragonesa ewes were controlled on the same farm and received intrauterine inseminations using the same technical equipment. The fertilizing ability of the raw semen and sperm samples selected by a dextran/swim-up process was compared using a low number of spermatozoa per insemination (7 x 10(7)) to enhance possible fertility differences. A new post-CCCD parameter was considered; the loss of viability (LV) occurred during the CCCD process. This variable denotes the sperm surviving ability and corresponds to the difference between the total number of viable cells loaded and recovered after the CCCD run. The mean fertility of eight sperm control samples was 60% (range: 25-76%), and there was no significant correlation between standard parameters and in vivo fertility. LV ranged from 2 to 69% (average 27%) and was negatively correlated with fertility (r = -0.914, P < 0.01). Ejaculate heterogeneity (H) ranged from 20 to 47% and was positively, but not significantly, correlated with fertility (r = 0.391). A predictive equation for fertility was deduced by multiple analysis with a very high correlation coefficient (r = 0.967), and level of significance (P < 0.005): predictive fertility PF = 52.546 - 0.594 LV + 0.665 H. The mean fertility of 13 swim-up selected samples was 63% (range: 25-86%). Again, only parameters derived from the CCCD analysis were highly correlated with fertility, especially LV and H (P < 0.05).

Animals↗

Strategic use of gonadotrophin-releasing hormone (GnRH) to increase pregnancy rate and reduce pregnancy loss in lactating dairy cows subjected to synchronization of ovulation and timed insemination.

The objective of this study was to determine the effect of GnRH (100 microg i.m.) treatment 5 and 15 days after timed insemination (TAI) on pregnancy rate and pregnancy loss in lactating dairy cows subjected to synchronization of ovulation. The study included 831 lactating dairy cows subjected to a Presynch-Ovsynch protocol for first service. On the day of TAI (Day 0), cows were randomly assigned to one of four experimental groups. Cows in Group 1 (n = 214) were treated with GnRH on Day 5; cows in Group 2 (n = 209) were treated with GnRH on Day 15; cows in Group 3 (n = 212) were treated with GnRH on both Day 5 and Day 15; cows in Group 4 (n = 196) were not treated. Pregnancy rate was evaluated at Day 27 and Day 45 after TAI. The interestrus interval and the proportion of cows diagnosed not pregnant based on expression of estrus and insemination before pregnancy diagnosis on Day 27 were determined. The results of this study are: (1) GnRH treatment on Day 5 or Day 15 did not increase pregnancy rate, or reduce pregnancy loss between Day 27 and Day 55 after TAI; (2) cows treated with GnRH on both Day 5 and Day 15 had a lower (P < 0.01) proportion of cows diagnosed not pregnant based on expression of estrus before ultrasonography on Day 27 (26.5%) compared to control cows (52.9%), and these cows had an extended (P = 0.05) interestrus interval (23.4 days vs. 21.5 days); and (3) GnRH treatment on both Day 5 and Day 15 after TAI reduced pregnancy rate on Day 27 (36.8% vs. 44.4% for control cows; P < 0.03) and Day 55 (28.3% vs. 36.2% for control cows; P < 0.01). Therefore, strategies to stimulate CL function using multiple doses of GnRH during the luteal phase need to consider potential negative effects.

Abortion, Veterinary↗

Deep freezing of concentrated boar semen for intra-uterine insemination: effects on sperm viability.

The use of deep-frozen boar semen for artificial insemination (AI) is constrained by the need for high sperm numbers per dose, yielding few doses per ejaculate. With the advancement of new, intra-uterine insemination strategies, there is an opportunity for freezing small volumes containing high sperm numbers, provided the spermatozoa properly sustain cryopreservation. The present study aimed to concentrate (2 x 10(9) spz/mL) and freeze boar spermatozoa packed in a 0.5 mL volume plastic medium straw (MS) or a multiple FlatPack (MFP) (four 0.7 mL volume segments of a single FlatPack [SFP]) intended as AI doses for intra-uterine AI. A single freezing protocol was used, with a conventional FlatPack (SFP, 5 x 10(9) spz/5 mL volume) as control. Sperm viability post-thaw was monitored as sperm motility (measured by computer-assisted sperm analysis, CASA), as plasma membrane integrity (PMI, assessed either by SYBR-14/PI, combined with flow cytometry, or a rapid hypo-osmotic swelling test [sHOST]). Sperm motility did not differ statistically (NS) between test-packages and control, neither in terms of overall sperm motility (range of means: 37-46%) nor sperm velocity. The percentages of linearly motile spermatozoa were, however, significantly higher in controls (SFP) than in the test packages. Spermatozoa frozen in the SFP (control) and MFP depicted the highest PMI (54 and 49%, respectively) compared to MS (38%, P < 0.05) when assessed with flow cytometry. In absolute numbers, more viable spermatozoa post-thaw were present in the MFP dose than in the MS (P < 0.05). Inter-boar variation was present, albeit only significant for MS (sperm motility) and SFP (PMI). In conclusion, the results indicate that boar spermatozoa can be successfully frozen when concentrated in a small volume.

Animals↗