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Effect on fertility of uterine lavage performed immediately prior to insemination in mares.

OBJECTIVE: To determine the effect on fertility of large-volume uterine lavage with lactated Ringer's solution (LRS) performed immediately prior to insemination in mares. DESIGN: Prospective randomized controlled study. ANIMALS: 20 mares. PROCEDURE: Control mares (n = 10) were inseminated with 1 billion (estimated before cooling) progressively motile spermatozoa that had been cooled in a passive cooling unit for 24 hours. Mares (n = 10) in the treatment group were inseminated with 1 billion progressively motile spermatozoa (cooled as described for control mares) immediately after uterine lavage with 4 L of sterile LRS. RESULTS: There were no significant differences in pregnancy rates or size of the embryonic vesicle on days 12, 13, and 14 after ovulation between control and treated mares. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that uterine lavage with LRS can be performed immediately prior to insemination without adversely affecting fertility in mares. This is clinically important, because insemination may be necessary when a mare has inflammation-associated fluid (detectable ultrasonographically) in the uterus; removal of the fluid is desirable, because it adversely affects spermatozoal motility and fertility. This situation typically arises when mares require rebreeding after they have developed persistent mating-induced endometritis or are inseminated multiple times in a 24-hour period (during the period of physiologic mating-induced inflammation), which is a common practice when using cooled or frozen-thawed semen.

Animals↗

Accessory sperm: their importance to fertility and embryo quality, and attempts to alter their numbers in artificially inseminated cattle.

Accessory sperm number and its relationship to fertilization and embryo quality was evaluated in cattle after nonsurgical recovery of ova or embryos 6 d after insemination. Efforts to alter accessory sperm number per ovum included 1) blockage of retrograde sperm loss at insemination using a modified insemination device, 2) elevated sperm number per inseminate (40 x 10(6) vs 20 x 10(6], and 3) alteration in semen quality (percentage of viable and morphologically normal sperm in the inseminate). None of these efforts affected accessory sperm number per ovum or embryo. However, blockage of retrograde semen flow for 3 h or use of semen of below-average quality (decreased percentage of viable and morphologically normal sperm) resulted in significant decreases in number of viable embryos and increases in number of degenerate embryos and unfertilized ova compared with conventional insemination (P less than .03) and use of semen with an average percentage of viable and morphologically normal sperm (P less than .06). Number of accessory sperm per embryo or ovum was positively related to fertilization and embryo quality (P less than .05). Mean accessory sperm +/- SD and the median value (in parentheses) for unfertilized ova, degenerate embryos, and embryos classified fair to poor and excellent to good were, respectively, .3 +/- .8 (0), 5.4 +/- 8.9 (1.0), 15.8 +/- 28.6 (3.5), and 16.9 +/- 29.5 (5.0). We conclude that efforts to improve accessory sperm numbers per embryo or ovum failed and that high variation and skewness of accessory sperm toward 0 may make median values more meaningful than means.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Artificial insemination outcomes in beef females using bovine sperm with a detectable fertility-associated antigen.

In this study, semen samples from 25 bulls that had passed a breeding soundness evaluation were analyzed for the presence or absence of a 31-kDa protein, known as fertility-associated antigen (FAA), on spermatozoal membranes. Eighteen bulls had FAA on sperm (FAA-positive) and seven were devoid of FAA on sperm (FAA-negative). A single ejaculate from each bull was extended and frozen with 25 to 30 x 10(6) sperm in .5-mL straws. Crossbred replacement heifers (n = 865) were estrus-synchronized and artificially inseminated either at timed AI or 12 h after they were detected in estrus. Mature cows (n = 285) were inseminated 12 h after they were detected in estrus during a 45-d AI period. Pregnancy rates (pooled) to first AI service for females (n = 764) inseminated with FAA-positive sperm were 65.6% and were 49.7% for females (n = 386) inseminated with FAA-negative sperm (P < .005). Among the estrus-synchronized replacement heifers, pregnancy rates to synchronized AI service for heifers (n = 550) inseminated with FAA-positive sperm were 62% and were 45.7% for heifers (n = 315) inseminated with FAA-negative sperm (P < .005). These data indicate that pregnancy rates to first AI service at spontaneous and synchronized estrus are higher when using semen from bulls with detectable FAA on spermatozoal membranes compared to semen from bulls devoid of FAA on membranes. Fertility-associated antigen is an important determinant for fertility potential of sperm from bulls to be used in AI breeding programs.

Animals↗

Effect of day of the estrous cycle at the initiation of a timed artificial insemination protocol on reproductive responses in dairy heifers.

Our objectives were to identify stages of the estrous cycle at which initiation of a timed artificial insemination (Ovsynch/TAI) protocol may reduce pregnancy rates and to monitor ovarian follicle dynamics and corpus luteum development after initiation of the Ovsynch/TAI protocol at different stages of the cycle. Cycling Holstein heifers (n = 24) were injected twice with prostaglandin F2alpha to induce estrus and were scanned by ovarian ultrasonography to determine the day of ovulation (d 0). Heifers were assigned to initiate the Ovsynch/TAI protocol at d 2 (n = 5), 5 (n = 5), 10 (n = 4), 15 (n = 5), or 18 (n = 5) of the cycle. The Ovsynch/TAI was initiated with an injection of gonadotropin-releasing hormone agonist followed 7 d later with an injection of prostaglandin F2alpha. At 36 h after injection of prostaglandin F2alpha, heifers were injected with gonadotropin-releasing hormone agonist and inseminated 16 h later. Heifers were scanned daily during the Ovsynch/TAI protocol and every other day after insemination until 16 d later. Blood samples were collected daily starting at the 1st day heifers were scanned and continued until 16 d after insemination. Initiation of the Ovsynch/TAI protocol at d 15 of the estrous cycle caused heifers to ovulate prior to insemination. A shortened return to estrus (< 16 d) was caused by ovulation failure to the second gonadotropin-releasing hormone injection, by incomplete regression of the corpus luteum, and by short life-span of the induced corpus luteum. Day of the cycle in which the Ovsynch/TAI protocol is initiated affects dynamics of follicular development, plasma progesterone profiles, and occurrence of premature ovulation. Size of the pre-ovulatory follicle was associated positively with subsequent progesterone concentrations following insemination.

Animals↗

Effect of time of oocyte collection and site of insemination on oocyte transfer in mares.

The objective of the study was to compare embryo development rates after transfer of oocytes collected 22 or 33 h after hCG injection into recipients inseminated within the uterus or the oviduct. Oocytes were collected at approximately 22 or 33 h after hCG injections and incubated for approximately 16 or 1.5 h, respectively, before transfer. Intrauterine inseminations using 1 x 10(9) progressively motile sperm were done approximately 12 h before and 2 h after transfer. For intraoviductal inseminations (gamete intrafallopian transfer [GIFT]), semen was centrifuged through a Percoll gradient, and 200,000 progressively motile sperm were transferred with oocytes into the oviduct. Time of oocyte collection (22 or 33 h) after hCG injection did not affect embryo development rates (17/25, 68%, vs 12/23, 52%, respectively; P = 0.40). When results from oocyte collections at 22 and 33 h after hCG were combined, oocyte transfer with intraoviductal vs intrauterine insemination resulted in similar (P = 0.70) embryo development rates (12/22, 55%, and 17/26, 65%, respectively). However, the interaction between time of oocyte collection and site of insemination tended to be significant (P = 0.09), suggesting that GIFT using oocytes collected at 33 h after hCG may not be as effective as using oocytes collected at 22 h after hCG. Because intraoviductal insemination requires a low number of sperm, GIFT could be used in cases of male subfertility, frozen semen, or sexed sperm.

Animals↗

Synchronization of Bos indicus x Bos taurus cows for timed artificial insemination using gonadotropin-releasing hormone plus prostaglandin F2alpha in combination with melengestrol acetate.

Nonlactating Bos indicus x Bos taurus cows were used in three herds to determine the efficacy of different PGF2alpha treatments in combination with GnRH and melengestrol acetate (MGA) for a timed artificial insemination protocol. The start of the experiment was designated as d 0, at which time cows were assigned a body condition score and received 100 microg of GnRH. Cows were fed MGA (0.5 x mg x cow(-1) x d(-1)) on d 1 to 7. On d 7, cows received either a single injection of PGF2alpha (Lutalyse sterile solution; 25 mg; n = 297), a single injection of cloprostenol sodium (Estrumate; 500 microg; n = 297), or half the recommended dose of PGF2alpha (12.5 mg; n = 275) on d 7 and 8. On d 10, all cows were artificially inseminated and received 100 microg of GnRH. Pregnancy rates to the timed artificial insemination (39%) were not affected by treatment, herd, or treatment x herd. There was an effect (P < 0.01) of artificial insemination sire on timed artificial insemination pregnancy rate for one herd, but not the other two herds. Herd influenced (P < 0.05) 30-d pregnancy rates, but there were no treatment or treatment x herd effects as 72.3% of the cows became pregnant during the first 30 d of the breeding season. Results indicate that the type of PGF2alpha treatment administered 7 d after GnRH did not influence timed artificial insemination pregnancy rates in nonlactating Bos indicus x Bos taurus cows.

Animals↗

Influence of estradiol-17 beta on fertility in confined sheep inseminated with frozen semen.

Impaired sperm transport is believed to be a major cause of reduced fertiity in ewes inseminated with frozen semen. Because of suggestions that estradiol improves sperm transport, estradiol-17 beta was given to mature ewes at progestagen-induced estrus to test effects on fertility after artificial insemination with frozen semen. Conception rates, based on progesterone determinations made 18 days after insemination, were 43% for ewes injected with 50 microgram estradiol-17 beta and inseminated with frozen semsn, 52% for ewes inseminated with frozen semen but given no estradiol and 86% for ewes inseminated with fresh semen. Lambing rates were 6, 35 and 69%, respectively. The markedly greater difference between conception rates and lambing rates for the estradiol-treated ewes suggests that estradiol caused a substantial increase in the early embryonic mortality associated with the use of frozen semen.

Animals↗

Increased numbers of sperm in the oviducts and improved fertilization rates in rabbits after administration of phenylephrine or ergonovine near the time of insemination.

Phenylephrine, an alpha-adrenoceptor agonist, was administered im to does near the time of mating or insemination. The treatment increased sperm numbers in the oviducts by about 50-fold and in the uterus by about 10-fold at 2 or 2.5 h after insemination. Methoxamine, another alpha-adrenoceptor agonist that was given im, did not increase sperm numbers, although both phenylephrine' and methoxamine significantly increased the number and amplitude of uterine contractions when contractions were measured by strain gauge force transducers attached to the uterus of conscious does. Ergonovine, an ergot derivative given im, increased sperm numbers more than 10-fold in the oviducts and five to 10-fold in the uterus at 2 or 2.5 h after insemination. Ergonovine increased the frequency and amplitude of uterine contractions when given iv but not when given im. In tests with a range of doses of phenylephrine and ergonovine, 5 mg of phenylephrine and .6 mg of ergonovine appeared to be near optimal for maximizing the number of sperm in the uterus and oviducts at 2.5 h after insemination. Phenoxybenzamine, an alpha-adrenoceptor blocking agent, prevented the phenylephrine-induced increases in both uterine contractions and sperm numbers in the oviducts and uterus. Phenoxybenzamine also prevented the effect of ergonovine on sperm numbers. In does inseminated with low numbers of sperm (92,000; an inseminate selected to result in a low fertilization rate in control does), the administration of phenylephrine or ergonovine significantly increased ovum fertilization rates (16% for control does, 52 and 63%, respectively, for phenylephrine- and ergonovine-treated does).

Animals↗

Time schedules of intrauterine insemination after urinary luteinizing hormone surge detection and pregnancy results.

A urinary luteinizing hormone (LH) test (LH Color, Organon, Oss, The Netherlands), was used to time intrauterine insemination in 177 cycles. Morning and evening urine samples were tested. In 58 women (33%) the test was positive in the morning urine sample. Fifteen of these patients were inseminated 8-10 h thereafter and one patient (6.7%) conceived. The remaining 43 women were inseminated the following day, 25-31 h after LH detection, and seven pregnancies (16.3%) ensued. In 119 cycles showing a positive urinary test in the evening sample, insemination was performed the next day, between 17 and 23 h after the LH surge, and 18 patients (15.1%) became pregnant. Statistical analysis showed no significant differences in pregnancy rates between the three different schedules, or in the time of insemination between conceptional and non-conceptional cycles within each group. Most ovulations occurred between 16 and 28 h after the positive test was observed. These findings suggest that while the lifespan of the gametes allows a relatively long period for fertilization, from 8 to 31 h after urinary LH surge detection, better results may be expected when inseminating about 24 (+/- 6) h after the positive test.

Adult↗

Use of estradiol cypionate in a presynchronized timed artificial insemination program for lactating dairy cattle.

Experiment 1 evaluated pregnancy rates when estradiol cypionate (ECP) was used to induce ovulation as part of a timed artificial insemination (TAI) protocol in comparison to Ovsynch for lactating dairy cows in Florida (n = 371) and Texas (n = 321). Cows were presynchronized with two injections of PGF2, (25 mg, im) given 14 d apart with TAI protocols beginning 14 d after the second injection of PGF20. The TAI protocols consisted of an injection of GnRH (100 microg, im) followed by PGF2alpha 7 d later. Then, cows either received an injection of GnRH (Treatment I, Ovsynch) at 48 h after PGF2alpha and inseminated 16 to 24 h later or received an injection of ECP (1 mg, i.m.) at 24 h after PGF2alpha, (Treatment II; Heatsynch) and inseminated 48 h later. In Florida, pregnancy rates after TAI were 37.1 +/- 5.8% for Ovsynch compared with 35.1 +/- 5.0% for Heatsynch. In Texas, pregnancy rates were 28.2 +/- 3.6% for Ovsynch and 29.0 +/- 3.5% for Heatsynch. Overall pregnancy rates did not differ between Ovsynch and Heatsynch treatments. In Experiment 2, estrus and ovulation times were determined in lactating dairy cows submitted to the Heatsynch protocol. Frequencies of detected estrus and ovulation after ECP were 75.7% (28/37) and 86.5% (32/37), respectively. Mean intervals to ovulation were 55.4 +/- 2.7 h (n = 32) after ECP and 27.5 +/- 1.1 h (n = 27) after onset of estrus. Estrus occurred at 29.0 +/- 1.8 h (n = 28) after ECP. It is recommended that any cow detected in estrus by 24 h after ECP injection be inseminated at 24 h and all remaining cows be inseminated at 48 h because 75% (n = 24/32) of the ovulations occurred between > or = 48 h to < or = 72 h after ECP. Synchronization of ovulation and subsequent fertility indicated that estradiol cypionate could be used to induce ovulation for successful timed insemination.

Animals↗

Short communication: presynchronization using a single injection of PGF2alpha before synchronized ovulation and first timed artificial insemination in dairy cows.

Conception to synchronized ovulation (Ovsynch) using injections of GnRH and PGF2alpha and timed artificial insemination has been shown to be maximized when the program is initiated 5 to 12 d after estrus. The objective of this double-blinded field trial was to assess the effect of one injection of PGF2alpha, 10 d before the Ovsynch program, on the probability of pregnancy at first insemination in lactating dairy cows. The hypothesis was that cows that underwent luteolysis in response to PGF2alpha would be between 5 and 8 d postestrus at the start of Ovsynch. In five commercial dairy herds in Ontario, Canada, at 52 +/- 12 d in milk, 506 cows were assigned at random to receive either one i.m. injection of 500 microg of cloprostenol or saline. Ten days later, all cows received 100 microg of GnRH i.m., followed in 7 d by 500 microg of cloprostenol i.m. and 100 microg of GnRH i.m. 48 h later. All cows were artificially inseminated 0 to 20 h after the second injection of GnRH, without regard to detection of estrus. Pregnancy was diagnosed by transrectal palpation at least 35 d after insemination. The probability of pregnancy after first insemination was modeled with logistic regression, accounting for the correlation of cows with herd and the effect of season of calving. There was no difference in pregnancy risk between cows that received PGF2alpha presynchronization and controls (37.3 and 36.6%, respectively; odds ratio = 1.03, 95% confidence interval, 0.88 to 1.20). Parity and days in milk at insemination were not significant covariates.

Animals↗

Number of inseminations to conception in Holstein cows using censored records and time-dependent covariates.

Three methodologies that accommodate censoring or time-dependent covariates were used to estimate variance components for number of inseminations to conception. Data included 80,071 lactation records and 143,927 artificial inseminations in 47,509 Spanish Holstein cows. Up to 4 inseminations to conception, along with their respective censoring information, were analyzed. An ordinal-censored threshold model (CTM), a sequential threshold model (STM), and a grouped survival analysis via a discrete proportional hazards model (DPH) were implemented. Sire variance estimates on the liability scale were 0.016 and 0.010 for CTM and STM, respectively, and 0.012 for DPH on the logarithmic scale. Heritability estimates on the liability scale were 0.050 and 0.038 with CTM and STM, respectively. All models led to similar rankings of sires, and the strong correlations (0.97 to 0.98) between methodologies suggested robustness in ranking of sires of cows. Service sire variance estimates were 0.021 for both CTM and STM; DPH led to an approximate service sire variance of 0.020. Rankings for service sires between methodologies ranged from 0.76 to 0.90. These lower values are most likely due to differences in the treatment of time-dependent covariates. The STM had greater predictive ability of daughter fertility at first insemination than the other methodologies. However, the CTM predicted daughter fertility more accurately in subsequent inseminations. The DPH and STM had a similar predictive ability of daughter fertility in second and subsequent inseminations.

Algorithms↗

Rates of conception by artificial insemination of dairy cattle.

Over 41 mo, 1004 first and second service inseminations were evaluated in Holstein cows and heifers of breeding age. Cattle were inseminated either at the end of the heat check in which they were observed in heat (0 h) or at the end of the next heat check (12 h). Rate of conception was affected by estrous behavior at 12 h, site of semen placement in the reproductive tract, sire, and environmental temperature the day after insemination. Service number, time of day at which insemination occurred, inseminator, and timing of insemination in relation to the onset of estrus had no effect on fertility. With twice daily heat checks, a single daily service produced acceptable rates of conception.

Animals↗

Controlling first service and calving interval by prostaglandin F2 alpha, gonadotropin-releasing hormone, and timed insemination.

Our objective was to determine if calving intervals could be shortened and made less variable by using prostaglandin F2 alpha to control the occurrence of first services. Holstein cows (n = 348) were assigned at calving to four treatment groups. Control cows (n = 88) were inseminated at their first observed estrus after 40 d postpartum. Estrous cycles of the remaining cows were synchronized with prostaglandin F2 alpha to allow insemination (first services) 80 h after the second injection (n = 86), insemination at 80 h preceded by gonadotropin-releasing hormone at 72 h (n = 86), or insemination at 72 and 96 h (n = 88) after the second injection (51 to 57 d postpartum). By design, interval to first service was reduced to 57 d for treated cows (63 d for controls) and was less variable (12% of that for controls). Conception rate at first service was lower after timed inseminations than that of controls. Intervals to conception and subsequent calving were similar in all treated cows and controls. Of cows sampled, 23 of 176 (13%) failed to respond with luteolysis when progesterone in serum exceeded 1 ng/ml, and 26 of 176 (15%) had low concentrations of progesterone in serum and could not respond to prostaglandin F2 alpha. Poor response to timed inseminations may have occurred because only 72% of 176 cows sampled responded with luteolysis. When only cows observed in estrus were considered, conception rate approached that of controls (51%).(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗

Nonreturn rates of dairy cattle following uterine body or cornual insemination.

In the dairy cattle industry, uncertainty still remains regarding the most desirable site of inseminate deposition to maximize AI conception rates. The effect of site of inseminate deposition on nonreturn rates was determined from 2195 cornual and 2428 uterine body-bred dairy cattle. Twelve technicians from various areas of Pennsylvania and New York were chosen on the basis of their accuracy of semen deposition in retraining sessions, average nonreturn rates, and their willingness to cooperate in the study. For a 3-mo period (June, July, and August 1988), technicians alternated weeks of cornual and uterine breeding on all dairy cattle inseminated. One-half (.25 ml) of each semen unit was deposited approximately 5.1 cm into each uterine horn for cornual insemination. No significant difference in nonreturn rates was found between horn-bred (70.8%) and body-bred (69.5%) cows. The range of differences in percent nonreturn rates for technicians was 19 and 30% for body and cornual inseminations, respectively. A significant difference in nonreturn rates was found between technicians and between months with significantly higher average nonreturn rates (6.8%) in June. Cornual and uterine body deposition of semen yielded similar results; therefore, depositing an inseminate in the uterine horns to maximize fertility is unnecessary.

Animals↗

The effect of artificial insemination once versus twice per day.

The objective of this study was to determine the effect of inseminating Jersey cows and heifers once per day or according to the a.m.-p.m. rule. A total of 337 artificial inseminations (AI) were completed by three technicians at the University of Tennessee Dairy Experiment Station at Lewisburg for 6 mo. Cows and heifers were inseminated at estrus using the a.m.-p.m. rule on even days of the month. On odd days of the month, AI were once daily between 0800 and 1200 h. Estrus detection was conducted two to three times daily. Pregnancy was confirmed by rectal palpation 60 to 80 d after AI. Herd DHIA averages were a 12.2-mo calving interval, 76 d to first AI, 83% observed estruses, and a 50% conception rate during the trial. Pregnancy data were analyzed with a model including treatment, AI, lactation number, parity, technician, and group. This study grouped cows and heifers according to when they were in estrus and inseminated (a.m.-a.m., a.m.-p.m., or p.m.-a.m.); means were 43.7, 57.9, and 59.0%, respectively. The a.m.-p.m. AI versus once per day AI yielded a pregnancy rate of 55.6% versus 51.3%. These results show no difference among Jersey cows or heifers that were inseminated artificially once daily in the a.m. However, those cows and heifers inseminated in the a.m. of first estrus detection had a lower pregnancy rate.

Animals↗

Sperm numbers inseminated in dairy cattle and nonreturn rates revisited.

Three experiments were conducted to test fertility when sperm numbers per insemination ranged from 10 x 10(6) to 40 x 10(6) total sperm. All semen was from Holstein bulls that were on a regular schedule of semen collection. The semen was extended with heated homogenized whole milk, cooled, glycerolated, and frozen according to standard procedures. Semen was distributed to a large group of inseminators to minimize differential field effects on treatment. All experiments were a randomized block design, including a split plot in Experiment 2. In Experiment 1, data for 31,399 first inseminations distributed among treatments of 20 x 10(6), 25 x 10(6), 30 x 10(6), and 40 x 10(6) total sperm resulted in 69.8, 70.0, 70.1, and 70.1% nonreturns at 59 d, respectively. In Experiment 2, data for 18,197 first inseminations divided over treatments of 12 x 10(6), 16 x 10(6), and 20 x 10(6) total sperm resulted in 70.2, 72.4, and 70.8% nonreturns at 59 d, respectively. In Experiment 3, 38,890 first inseminations distributed over treatments of 10 x 10(6), 13 x 10(6), 16 x 10(6), and 20 x 10(6) total sperm resulted in 70.5, 72.2, 73.1, and 71.5% nonreturns at 59 d, respectively. Bull nonreturns ranged from 64 to 76% in the three trials. These results indicate that, under good conditions, total sperm numbers per straw can be reduced to 10 x 10(6) total sperm with a reduction of nonreturn rates at 59 d, for most bulls, of about 1 percentage unit from the maximum when professional inseminators are use.

Animals↗

A revised artificial insemination schedule for broiler breeder hens.

Broiler type hens maintained in individual cages were artificially inseminated with either 0.023, 0.035, or 0.047 ml. of pooled semen. Insemination intervals were nine, nine, and ten days on a repetitive basis. This schedule was adopted because it allowed an extension of the conventional insemination interval and yet remained compatible with a five-day industrial work-week. The results demonstrated that maintaining broiler breeder hens in cages and utilizing artificial insemination as a means of obtaining fertile eggs were feasible. The data also indicated that extending the insemination interval to 10 days is possible providing the number of spermatozoa inseminated is increased.

Animals↗