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PubMed1986

Removal of bacterial contaminants from semen for in vitro fertilization or artificial insemination by the use of buoyant density centrifugation.

Buoyant density centrifugation of semen produces the accumulation of populations of highly motile, morphologically normal spermatozoa in the lowermost 1 ml of Percoll (Pharmacia Fine Chemicals AB, Uppsala, Sweden) density gradients. In addition, the majority of bacteria present in semen are retained in the seminal plasma at the top of the gradients. Of 40 semen samples examined, 37 contained detectable bacteria, but after buoyant density centrifugation, the spermatozoal populations collected from the lowermost 1 ml of the Percoll columns were found to contain few or no bacteria. When preparations were collected using sterile technique (by boring a hole through the bottom of the centrifuge tube), 14 of the 20 preparations were found to be bacteria-free. When preparations were collected by passing a spinal needle from the surface through the seminal plasma to the bottom of the centrifuge tube, the sterility of the final spermatozoa preparations was not maintained, with only 5 of the 20 samples completely free of bacteria. The residual bacterial contamination of the remaining 15 samples was, however, very low (less than 5 colonies after a 48-hour culture period).

Bacteria↗
PubMed1993

Comparison of the methods of artificial insemination on the incidence of conception in single unmarried women.

OBJECTIVE: To compare pregnancy rates after intrauterine insemination (IUI) versus pericervical insemination in absolute male factor infertility using each patient as her own control. DESIGN: Ovulatory women with patent fallopian tubes without male partners were alternately inseminated with cryopreserved donor semen using either IUI or pericervical insemination techniques. A total of 81 cycles, which included up to 4 cycles per patient were performed. In this manner a comparison between the efficacy of each method could be evaluated. SETTING: The donor insemination program at the Center For Assisted Reproduction at Northwestern University Medical School. PATIENTS: Twenty-six single, healthy, unmarried women with patent fallopian tubes and < 40 years of age without male partners (absolute male factor infertility). MAIN OUTCOME MEASURES: Positive quantitative serum subunit of human chorionic gonadotropin followed by the presence of an intrauterine gestational sac seen by transvaginal ultrasonography. RESULTS: Fourteen (54%) of 26 patients conceived including two (14%) miscarriages within four insemination cycles. Seven (17.5%) patients after IUI, and 7 (17.1%) patients after pericervical insemination conceived. The pregnancy rates were similar regardless of the order of insemination method. CONCLUSION: These findings reveal that there is no statistical difference in the pregnancy outcome between these two methods of insemination in absolute male factor infertility.

Adult↗
PubMed1994

Sperm-immobilizing antibodies in women undergoing artificial insemination with donor sperm.

Forty-nine female partners of infertile couples undergoing AID (1 to 44 times) were tested for the presence of sperm-immobilizing antibodies in their serum. One hundred fifty-one infertile women undergoing ART treatment other than AID were enrolled as a control group. One of the 49 AID patients and 3 of the 151 women of the control group were positive for sperm-immobilizing antibodies. The sperm-immobilizing antibodies incidence in AID patients did not differ significantly from that in the control group. This suggests that multiple exposures to semen from different donors do not increase the possibility of inducing sperm-immobilizing antibodies in AID patients.

Antibodies↗
PubMed1996

Risks for transmission of hepatitis C virus during artificial insemination.

OBJECTIVE: To identify risks of hepatitis C virus transmission by semen from infected donors. DESIGN: Case report. SETTING: Assisted fertility clinic. PATIENTS: Hepatitis C virus-infected semen donor and recipients of his donations. INTERVENTION: Testing for hepatitis C virus by serology and polymerase chain reaction. MAIN OUTCOME MEASURES: Detection of hepatitis C virus antibodies and viral RNA. RESULTS: Hepatitis C virus RNA was detected in the semen donation before but not after purification; none of the recipients of the donors samples were found to have antibodies to hepatitis C virus. CONCLUSIONS: Hepatitis C virus RNA can be detected in semen donations from infected donors; purification of donations before insemination significantly reduces the amount of viral RNA in the semen pellet.

Antibodies, Viral↗
PubMed1988

Spermatozoa selection in discontinuous Percoll gradients for use in artificial insemination.

In a preliminary study of the ejaculate of 93 men, changes in motility and hypo-osmotic swelling before and after sperm separation by discontinuous Percoll gradients were evaluated. Both parameters improved significantly (P less than 0.01) in Percoll-separated spermatozoa. In a second stage, 99 couples underwent intrauterine insemination of separated semen by Percoll gradients. The population had infertility for a mean of 5.6 years. In a 1-year period, a total of 344 intrauterine insemination cycles were completed. Failure was considered when conception did not occur within four treatment cycles. The mean number of cycles per successful couple before pregnancy was 1.52. An overall 21% pregnancy rate was achieved (16% for oligoasthenospermia and 24% for asthenospermia). Sperm separation in Percoll gradients is a useful technique for intrauterine insemination in some cases of male subfertility.

Adult↗
PubMed2000

Induction of follicular wave emergence for estrus synchronization and artificial insemination in heifers.

The objective was to synchronize follicular wave emergence among cattle for synchronization of estrus and ovulation, and to determine pregnancy rate after AI at observed estrus. At random stages of the estrous cycle, a controlled internal drug release device (CIDR-B) was inserted intravaginally (Day 0) in 67 cross-bred beef heifers, and they were randomly allocated to receive either no further treatment (Control; n = 18); 5 mg of estradiol-17beta and 100 mg of progesterone im (E/P; n = 16); 100 microg im of GnRH (GnRH; n = 16); or transvaginal ultrasound-guided follicular ablation of all follicles > or = 5 mm (FA; n = 17). All heifers received a luteolytic dose of PGF (repeated 12 h later), and CIDR-B were removed on Days 9, 8, 6 or 5, in Control, E/P, GnRH or FA groups, respectively, so the dominant follicle of the induced wave was exposed to exogenous progesterone for a similar period of time in each group. Mean (+/- SEM) intervals (and range, in days) from treatment to follicular wave emergence in these groups were 3.5 +/- 0.6 (-2 to 8), 3.4 +/- 0.1 (3 to 4), 1.5 +/- 0.3 (-1 to 4), and 1.0 +/- 0.1 (0 to 2), respectively. Although the interval was longest (P<0.01) in the E/P and Control groups, it was least variable (P<0.01) in the E/P and FA groups. Intervals (and range, in days) from CIDR-B removal (and first PGF treatment) to estrus were 2.3 +/- 0.2 (1.5 to 4.5), 2.2 +/- 0.2 (1.5 to 3.0), 2.1 +/- 0.1,(1.5 to 3.5), and 2.5 +/- 0.1 (2.0 to 3.5), and to ovulation were 3.5 +/- 0.2 (2.5 to 5.5), 3.4 +/- 0.1 (3.0 to 4.5), 3.5 +/- 0.1 (2.5 to 4.5), and 3.8 +/- 0.1 (3.0 to 4.5), for Control, E/P, GnRH and FA groups, respectively (ns). The proportion of heifers displaying estrus was higher in the Control than in the FA group (94% versus 65%, P<0.05) and intermediate in EP and GnRH groups (87% and 75%). Heifers were inseminated approximately 12 h prior to ovulation (based on estrous behavior and ultrasound examinations). Pregnancy rates were 78%, 80%, 69% and 65% for Control, E/P, GnRH and FA groups, respectively (P=0.73). Results support the hypothesis that synchronous follicular wave emergence results in synchronous follicle development and, following progesterone removal, synchronous estrus and ovulation with high pregnancy rates to AI. The synchrony of estrus and ovulation in the E/P, GnRH and FA groups suggest that these treatments, in combination with CIDR-B, could be adapted to fixed-time insemination programs.

Animals↗
PubMed2001

Ram-induced ovulation to improve artificial insemination efficiency with frozen semen in sheep.

Ram effect, defined as shortening of seasonal anestrus in ewes by exposure to the ram, is now well recognized but the underlying mechanisms are still unclear. Little information also exists whether the ram is able to influence the estrus cycle and ovulation. Three experiments were conducted to investigate endocrine response, time of ovulation and pregnancy rate of ewes in proestrus, exposed to the ram (treated) or an adult ewe (control). In the first experiment, ewes (n = 20) were treated with fluorgestone acetate pessaries for 12 days and were given eCG and cloprostenol one day before withdrawal of pessaries. On the day after removal of the pessaries ewes in the treated group (n = 10) were exposed to the ram and those in the control group (n = 10) were exposed to an adult ewe. Blood samples were taken for LH assay every 20 min from 2 h before to 24 h after ram exposure. In the second experiment, ewes (n = 120) were induced into proestrus and on the day after removal of the pessaries were exposed to either a ram (n = 60) or a ewe (n = 60) as described above and were laparoscoped 50, 60 or 70 h after pessary withdrawal (n = 20 at each time interval). In the third experiment ewes (n = 90) were induced and exposed to the ram (n = 45) or an adult ewe (n = 45) and inseminated via a laparoscope whit frozen-thawed semen at 50 or 60 h after pessary removal, respectively. Exposure to the ram was followed in 2 h by a marked rise in LH, equivalent to a preovulatory surge in duration and amplitude. It was also followed by concentrated ovulation within 25 to 30 h and by an increased pregnancy rate in exposed ewes (73.3 vs. 53.3%).

Anestrus↗
PubMed2002

The use of progestins in regimens for fixed-time artificial insemination in beef cattle.

Four experiments were conducted to investigate modifications to gonadotropin releasing hormone (GnRH)-based fixed-time Al protocols in beef cattle. In Experiment 1, the effect of reducing the interval from GnRH treatment to prostaglandin (PGF) was examined. Lactating beef cows (n = 111) were given 100 mg gonadorelin (GnRH) on Day 0 (start of treatment) and either 500 microg cloprostenol (PGF) on Day 6 with Al and 100 microg GnRH 60 h later, or PGF on Day 7 with Al and GnRH 48 h later (6- or 7-day Co-Synch regimens). Pregnancy rates were 32/61 (53.3%) versus 26/50 (52.0%), respectively (P = 0.96). In Experiment 2. cattle (n = 196) were synchronized with a 7-day Co-Synch regimen and received either no further treatment or a CIDR-B device (Days 0-7). Pregnancy rates were 32/71 (45.1%) versus 33/77 (42.9%) in cows (P < 0.8), and 9/23 (39.1 %) versus 17/25 (68.0%) in heifers (P < 0.05). In Experiment 3, 49 beef heifers were randomly assigned to receive 12.5 mg pLH on Day 0, PGF on Day 7 and 12.5 mg of pLH on Day 9 with Al 12 h later (pLH Ovsynch), or similar treatment plus a CIDR-B device from Days 0 to 7 (pLH Ovsynch + CIDR-B), or 1 mg estradiol benzoate (EB) and 100 mg progesterone on Day 0, a CIDR-B device from Days 0 to 7 (EB/ P4 + CIDR-B), PGF on Day 7 (at the time of CIDR-B removal) and 1 mg i.m. EB on Day 8 with AI on Day 9 (52 h after PGF). Pregnancy rate in the EB/P4 + CIDR-B group (75.0%) was higher (P < 0.04) than in the pLH Ovsynch group (37.5%): the pLH Ovsynch + CIDR-B group was intermediate (64.7%). In Experiment 4, 266 non-lactating cows were allocated to a 7-day Co-Synch protocol (Co-Synch), a 7-day Co-Synch plus 0.6 mg per head per day melengestrol acetate (MGA) from Days 0 to 6 inclusive (Co-Synch + MGA) or MGA (Days 0-6) plus 2 mg EB and 50 mg progesterone on Day 0. 500 microg PGF on Day 7, 1 mg EB on Day 8 and fixed-time Al 28 h later (EB/ P4 + MGA). Pregnancy rates (P < 0.25) were 44.8% (39/87: Co-Synch), 47.8% (43/90; Co-Synch + MGA), and 60.7% (54/89: EB/P4 + MGA). In conclusion, a 6- or 7-day interval from GnRH to PGF in a Co-Synch regimen resulted in similar pregnancy rates in cows. The addition of a progestin to a Co-Synch or Ovsynch regimen significantly improved pregnancy rates in heifers but not in cows. Progestin-based regimens that included EB consistently resulted in high pregnancy rates to fixed-time Al.

Animals↗
PubMed1999

A preliminary study on the usefulness of huIL-8 in cervical relaxation of the ewe for artificial insemination and for embryo transfer.

The efficacy of using human interleukin 8 (huIL-8) as an agent for inducing cervical relaxation in estrous and diestrous sheep was assessed in a small pilot study. Multiparous, estrus-synchronized ewes were treated for either 2 or 5 consecutive days with vaginal suppositories with or without 5 micrograms cytokine. Cervical penetration with an insemination instrument was then assessed in vivo. After euthanasia, physical, histological and enzymological properties of the cervix were examined. Treatment of diestrous sheep with huIL-8 did not result in recruitment of neutrophils into the cervix. Treatment of estrous sheep with huIL-8 usually led to neutrophil recruitment to the cervix and to either full or partial penetration of the cervix. However, some animals receiving placebo treatment had neutrophil infiltration of both the vagina and cervix and, in one of these, partial penetration of the cervix was also achieved. Thus, treatment with IL-8 as the sole agent in the vaginal suppository was not sufficient to relax the cervix of the nonpregnant ewe in this study.

Animals↗
PubMed1999

Effect of cooling rates on post-thaw sperm motility, membrane integrity, capacitation status and fertility of dairy bull semen used for artificial insemination in Sweden.

We studied the effects of 2 different cooling rates during equilibration of semen from room temperature to 4 degrees C, at 4.2 degrees C/min (control split sample) or at 0.1 degree C/min (treatment split sample) on in vitro sperm viability post thawing and fertility after AI. Forty batches of split-frozen semen from 14 dairy bulls (Swedish Red and White breed) aged 14 to 16 m.o. or 66 to 79 m.o. were evaluated post-thawing for sperm motility (visual and computer-assisted sperm analysis [CASA], membrane integrity (fluorescent microscopy and flow cytometry post-loading with the combined fluorophores Calcein AM/EthD-1 and SYBR-14/PI); acrosomal status (with Pisum sativum agglutinin [PSA] staining); and capacitation status (CTC-assay). Fertility values (56-d nonreturn rate) of the slow cooling batches (treatment) were 0.4% units higher than for faster cooled (control) batches, but the difference was not statistically significant. Fertility values for the older bulls were 1.6% units higher than for the group of younger sires. No statistically significant correlations were found between semen viability parameters assessed in vitro and 56-d nonreturn rate. Visually assessed sperm motility, membrane integrity, capacitation and acrosomal status post-thawing did not differ significantly between cooling procedures, however the percentage of motile spermatozoa and the kinetic characteristics of spermatozoa--average path velocity (VAP), straight path velocity (VSL) and curvilinear velocity (VCL)--assessed by CASA differed significantly between cooling procedures. The results indicate that most of the in vitro sperm viability parameters post-thawing and the fertility results for bulls after AI did not differ significantly between the 2 semen cooling procedures tested.

Acrosome Reaction↗
PubMed1999

Effect of timing of artificial insemination on sex ratio.

For a number of years, the time of insemination or mating during estrus has been believed to influence the sex ratio of offspring, with early insemination resulting in more females and late insemination, more males. Possible mechanisms of altering the sex ratio include facilitating or inhibiting the transport of either X- or Y-chromosome-bearing sperm through the reproductive tract, preferential selection of sperm at fertilization, or sex-specific death of embryos after fertilization. In livestock species, there is evidence for preferential selection of X- or Y-bearing sperm, based on the maturational state of the oocyte at fertilization. In deer and sheep, early and late insemination appears to skew the sex ratio toward females and males, respectively. In cattle, conflicting reports on the effect of time of insemination on sex ratio make the premise less clear. Many of the published studies lack adequate observations for definitive conclusions and/or are based on infrequent observations of estrus, making it difficult to assess the effect of time of insemination on sex ratio. It is likely that any effect of time of insemination on sex ratio in cattle is relatively small. Evidence is accumulating that treatments used for synchronization of estrus or ovulation in cattle may influence the sex ratio.

Animals↗