Fertilization rates in superovulating cows after deposition of semen on the infundibulum, near the uterotubal junction or after insemination with high numbers of sperm

Three experiments were conducted with 105 superovulating Holstein dairy cows in attempts to improve the fertilization rate. Cows were superovulated with follicle-stimulating hormone (FSH) and time of estrus was regulated with prostaglandin F(2)alpha (PGF(2)alpha). Semen was deposited on each infundibulum through a laparoscope inserted through the flank (Experiment 1) or near the uterotubal junctions through flexible tubing passed through the cervix and uterine horns (Experiment 2). In the third experiment, high numbers of sperm in fresh semen were deposited in the uterus. Cows were necropsied and ova were recovered and examined about 3.5 d after the beginning of estrus. Deposition of 0.5 ml of frozen-thawed semen on each infundibulum (Experiment 1) reduced both ovum recovery and fertilization. In ten cows inseminated on the infundibulum, ova representing 43% of ovulation points were recovered and 9% of these recovered ova were fertilized. In ten control cows, ova representing 80% of ovulation points were recovered and 62% of them were fertilized. In a 2 x 2 experiment with 36 superovulating cows (Experiment 2), 1 ml of diluted fresh or frozen semen was deposited either near the uterotubal junction or in the uterine body. The overall fertilization rate was 61%, with no significant effect of site of semen deposition or type of semen used. In Experiment 3, 2 or 3 ml of neat semen (average of 4.4 billion sperm) was deposited in the uterus of 12 cows; 183 of 197 intact ova (93%) were fertilized. In 56 control cows inseminated with 0.5 to 1.5 ml of frozen diluted semen (average of 70 million sperm), 502 of 947 intact ova were fertilized (53%, P<0.001). Insemination with high numbers of fresh sperm overcame problems of sperm loss or sperm transport and improved the fertilization rate.     

A simplified fertility test in the dairy bull utilizing superovulated dairy cows

Dairy bull fertility level has received less attention than production transmitting ability. A simplified fertility test may be beneficial. A study was designed to test the use of tris-(1-aziridinyl)-phosphine oxide (TEPA) treated sperm, which arrests early cell division of the fertilized egg, in heterospermic insemination of superovulated cows. Semen samples were collected and pooled from University of Illinois dairy bulls. Semen samples were washed once, suspended in Illini Variable Temperature diluent (IVT) and incubated with or without TEPA (1.0 to 5.0 mg/ml) for 15 min. Samples were then washed again to remove excess TEPA. Additions of 1.0 to 5.0 mg/ml TEPA to sperm concentrations of 8 x 10(8) sperm/ml had no adverse effect on motility or morphology. The first part of the study utilized superovulated cows inseminated with treated (six cows) or untreated (six cows) sperm in different samples from the same bulls. Secondly, superovulated cows (eight cows) were artificially inseminated with treated and untreated split ejaculates from the same bulls. Lastly, superovulated cows (five cows) were heterospermically inseminated with treated (bull No. 1) and untreated (bull No. 2) spermatozoa. Out of 54 and 39 ova recovered in control and test cows, 40 blastocysts and 31 embryos arrested at the one- to five-cell stage resulted, respectively. Out of a predicted 123 ovulations, 78 fertilized ova were recovered; 40 of these were fertilized by control spermatozoa and 36 by TEPA-treated spermatozoa for parts one and two of the study respectively. These results indicated no significant difference in fertilizability of ova between control and TEPA-treated spermatozoa. Of 41 fertilized ova recovered (part 3), bull No. 1 fertilized significantly more ova (mean +/- standard deviation 5.0 +/- 2.3) than bull No. 2 (2.6 +/- 1.8). Results indicate a difference in fertility between bulls.     

Relationship of sperm migration in anterior vaginal fluid and embryo quality in superovulated dairy heifers

Anterior vaginal fluid samples from 26 superovulated dairy heifers at insemination were classified into 2 grades: 1) with abnormal sperm penetration ability when vanguard sperm migration was randomly oriented; and 2) with normal sperm penetration ability when vanguard sperm migration was parallel and unidirectional. Vanguard sperm behavior and vanguard sperm migration distances were evaluated for their effects on fertilized ova recovery rates and transferable embryo recovery rates. Twelve vaginal fluid samples (46%) showed abnormal sperm penetration ability (Grade 1) and the mean +/- sem distance traveled by the vanguard spermatozoa into these samples (7.3 +/- 0.9 mm) was different (P<0.0001) from the remainder of the samples (48 +/- 3.4 mm) in which sperm penetration was registered as normal (Grade 2). In heifers in which anterior vaginal fluid samples were Grades 1 and 2, the fertilized ova recovery rates were 78.3% and 80.2%, respectively (P=0.91). Transferable embryo recovery rates were 54.2% and 32% for Grades 1 and 2, respectively (P<0.001). Using correlation and linear regression analysis, the sperm migration distance in Grade-2 samples was not related to fertilized ova (P=0.77) or to transferable embryo recovery rates (P=0.97). These results indicate that vanguard sperm movement into capillary tubes varied qualitatively and was related to subsequent embryo quality.     

In vitro fertilization in the rabbit after delayed ovum recovery

Rabbit ovum donors were superovulated with pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Ova were recovered 16-17 h post-hCG from oviducts immediately after killing and from excised oviducts held in saline 30 min at 33 degrees or 38 degrees C prior to ovum recovery. In vivo-capacitated spermatozoa were used to inseminate both groups of ova. Data revealed a decrease in fertilization rates following a 30-min delay at 38 degrees C in ovum recovery. Thus, 64% (44/69 ova) were fertilized with rapid recovery, whereas 43% (39/90 ova) were fertilized following a 30-min delay. The decrease in fertilization imposed by delay in ovum recovery was apparently overcome when oviduct storage was at 33 degrees C. Under these conditions, 69% of inseminated ova were fertilized. Ova inseminated with in vitro-capacitated sperm showed a similar response to delayed ovum recovery. Embryonic development in culture of ova obtained from mated does was not affected by delay in recovery at 33 degrees or 38 degrees C provided mated does had been injected only with hCG. Ova from mated does receiving both PMSG and hCG were adversely affected by a 38 degrees C delay. The data emphasize the importance of rapid ovum recovery from oviducts and suggest the possibility of altering conditions to overcome damaging effects of delayed recovery.     

Can spermatozoa with abnormal heads gain access to the ovum in artificially inseminated super- and single-ovulating cattle?

The collective efficiency of barriers in the female tract against spermatozoa with abnormal heads was studied. In Experiment 1, Day 6 ova/embryos were recovered nonsurgically from superovulated (n = 24) and single-ovulating (n = 44) cows following artificial insemination with semen of bulls selected for normal spermatozoal motility (> or = 50%) and high content (> 30%) of spermatozoa with misshapen heads, random nuclear vacuoles or the diadem defect. To assess characteristics of spermatozoa capable of traversing barriers in the female tract, accessory spermatozoa were classified morphologically (x 1250) and compared with those of the inseminate. Superovulated cows proved inadequate for assessment of accessory spermatozoa due to evidence of poor sperm retention in the zona pellucida; thus, only single-ovulating cows were used. Accessory spermatozoa (n = 479) from 31 ova/embryos recovered from 44 cows were more normal in head shape than those in the inseminate (76 vs 62%; P < 0.05). Spermatozoa with normal head shape, but with nuclear vacuoles appeared as accessory spermatozoa at the same frequency as they were found in the inseminate (20 vs 17%, respectively). Only sperm cells with subtly misshapen heads appeared as accessory spermatozoa. In Experiment 2, semen pooled from 4 bulls having large numbers of spermatozoa exhibiting a gradation from severely asymmetrically misshapen heads to subtly misshapen heads was evaluated. Again, the accessory sperm population (960 sperm cells recovered from 64 ova/embryos) was enriched with spermatozoa of normal head shape relative to the inseminate (53 vs 26%, respectively; P < 0.05). Sperm cells with only nuclear vacuoles and those with subtly misshapen heads were not different between the accessory and inseminate populations (11 vs 8%, and 20 vs 25%, respectively). We conclude that morphologically abnormal spermatozoa are excluded from the accessory sperm population based upon severity of head shape distortion.     

Efforts to correlate laboratory with field observations on bull sperm fertility

This work represents efforts towards development of the zona-free hamster ovum sperm penetration assay for predicting relative levels of fertility for semen from individual bulls. Results reported here followed insemination of hamster vitelli with bull sperm, after frozen storage, with observations of sperm acrosomes and parallel inseminations of more than 1000 cows with semen from each bull. The average 75-day non-return rate for the four bulls was 74.0% (range 71.6 to 75.6). Laboratory studies indicated the following: the percentage of sperm with intact acrosomes varied from 55 to 73 between bulls, the percentage of motile sperm varied from 41 to 64, the percentage of sperm with progressive motility ranged from 24 to 40, the number of sperm interacting per (zona-free hamster) ovum ranged from 1.6 to 3.8, the number of sperm attached per ovum ranged from 1.4 to 2.9, the number of sperm within each penetrated ovum ranged from 1.5 to 1.8, the percentage of ova interacting with sperm ranged from 76 to 92, the percentage of ova penetrated ranged from 62 to 85, and the percentage of ova with male pronuclei ranged from 33 to 49. Although predictive ranking in the laboratory of these bulls with less than 4% variation in fertility levels was not possible, the zone-free hamster ovum test could be useful in identifying potentially subfertile bulls before they enter a young sire-sampling program.     

Evidence for ovulation and fertilization in beef cows with short estrous cycles

Calves were weaned from 15 Polled hereford anestrous cows 25 to 42 days after calving. In eight cows the uterus was flushed on day 6 or 8 after the first postweaning estrus (day 0), and in seven cows the oviduct ipsilateral to the ovary containing an ovulation papilla was removed and flushed on day 3. One ovum (morula) was recovered from the eight uterine flushings, while six ova were recovered from six of the seven oviductal flushings. Of the six, three were fertilized (4 to 8 cells), two unfertilized and only the broken zona pellucida of one was recovered. An ovulation papilla was observed in all cows at the time of oviduct removal. Six of the 15 cows had cycles less than 12 days, and from four of those six fertilized ova were recovered. The data indicate that previously anestrous cows ovulate at their first postweaning estrus and the ova released are capable of being fertilized. Failure to maintain pregnancy appears to be due to early corpus luteum regression.     

Recovery rates and embryo quality following dominant follicle ablation in superovulated cattle

To determine the association between dominant follicle ablation and the outcome of a superovulatory regimen, two data sets were constructed from records of 171 recoveries from non-ablated cows and 1214 recoveries from cows that underwent follicular ablation prior to FSH treatment. Data set 1 included all cows with 2 or more records (n = 1385). Data set 2 included paired data for 87 cows which had at least 2 records of both ablated and non-ablated superovulatory attempts. Dominant follicle ablation was performed by use of transvaginal, ultrasound guided aspiration 48 hr prior to the start of FSH. The same FSH protocols were used for both ablated and non-ablated cows. For all cows (data set 1), more total ova/embryos were recovered from the ablation group (12.1+/-0.3 vs 10.5+/-0.8; P=0.06). This difference could be accounted for by greater numbers of non-transferable embryos in the ablation group (6.5+/-0.2 vs 5.3+/-0.6; P>0.01). For the paired data (data set 2), greater numbers of total ova/embryos recovered from the ablation group (12.8+/-1.0 vs 9.7+/-0.7; P=0.01) could also be accounted for by higher numbers of nontransferable embryos in this group (7.8+/-0.8 vs 4.5+/-0.4; P>0.01). There were no differences between groups for high quality embryos, percent cows producing no ova/embryos or percent cows producing no transferable embryos. These data support the premise that synchronization of follicular waves following dominant follicle ablation increases total ova/embryo output. However, the additional embryos were primarily nontransferable thereby negating potential economic gains.     

Fertilization of rabbit ova and the role of ovum investments in the block to polyspermy

The rabbit ovum seldom becomes polyspermic despite the presence of supernumerary sperm with easy access to the ovum plasma membrane. The purpose of this study was to characterize the role of ovum investments in blocking polyspermy. Ova were inseminated with capacitated sperm in vitro and were fertilized. Early stages of development were normal. The incidence of polyspermy was determined by cytological examination of fixed ova. The incidence of polyspermy increased following removal of both the zona pellucida and corona radiata but not following removal of only the corona radiata. These results suggest that the failure of supernumerary sperm to penetrate the ovum plasma membrane is at least in part due to the presence of the zona pellucida.     

Comparison of sperm binding potential of uninseminated, inseminated-unfertilized, and fertilized-noncleaved human oocytes under hemizona assay conditions.

In this study, human oocytes obtained after ovarian hyperstimulation for in vitro fertilization (IVF) and gamete intrafallopian transfer (GIFT) were utilized to evaluate sperm/zona pellucida binding potential. Three groups of oocytes were evaluated: 1) uninseminated; 2) inseminated-unfertilized; and 3) fertilized-uncleaved. All oocytes had undergone germinal vesicle breakdown at the time of retrieval and were salt-stored (pH 7.2) for not more than 30 days. Sperm binding was recorded under hemizona assay (HZA) conditions using spermatozoa from eight fertile men (HZA control) and from 1) four teratozoospermic (HZA test) and 2) four normozoospermic (HZA test) infertile men. First, the mean numbers (+/- SD) of sperm tightly bound for fertile controls and teratospermic men to hemizonae from uninseminated oocytes were 69.7 +/- 16 and 14.5 +/- 7, respectively (P = 0.02). Likewise, hemizonae from uninseminated oocytes bound 102.0 +/- 19 and 114.0 +/- 28, respectively, for fertile controls and normospermic men (P = 0.5). Second, hemizonae obtained from inseminated-unfertilized IVF oocytes bound 44.2 +/- 12 and 19.7 +/- 6 for fertile controls and teratospermic men, respectively (P = 0.02). This category of oocytes bound 100.5 +/- 7 and 108.5 +/- 11 sperm, respectively, for fertile controls and normospermic semen (P = 0.3). Third, HZA results of fertilized but uncleaved oocytes showed a mean number of tightly bound sperm of 6.0 +/- 4 compared with 65.0 +/- 1 in control, uninseminated oocytes using fertile sperm. These results demonstrate that uninseminated and inseminated-unfertilized human oocytes, salt-stored under controlled pH conditions, give reliable information regarding sperm binding potential under HZA conditions.     

Embryo production and quality of Holstein heifers and cows supplemented with beta-carotene and tocopherol

The hypothesis was that the intramuscular injection (i.m.) of beta-carotene associated to tocopherol improves cow (n=86) and heifer (n=91) embryo production and quality. Time of estrus was synchronized in animals with an ear implant with 3 mg of norgestomet associated with an i.m. injection of 6 mg of norgestomet and 10mg of estradiol valerate (CRESTAR, Intervert International B.V., Boxmeer, Holland) and superovulated by 8 i.m. FSH/LHp injections (400 IU-heifers and 500 IU-cows) in decreasing concentrations at 12h intervals. Animals were inseminated 12 and 24h after observed onset of estrus and embryos recovered 7 days later. Animals were randomly allocated to one of three treatments: (1) vegetable oil vehicle (control), (2) 800 mg of beta-carotene and 500 mg of tocopherol (T800) and (3) 1200 mg of beta-carotene and 750 mg of tocopherol (T1200). Supplemental injections were given at the day norgestomet implants were inserted and at first superovulatory injection. An index (Embryo Quality Index or EQI) was proposed to more precisely evaluate embryo quality (excellent*1 + good*2 + regular*3 + poor*4 + degenerate*5 + unfertilized ova*5)/total. There was an interaction between physiological stage (heifer or cow) and treatment on EQI (P=0.01) and on the proportion of viable embryos (P=0.03), where both variables were improved in T1200 cows, but not in heifers. The average EQI for heifers and cows in control, T800 and T1200 were 2.6+/-0.3 and 3.6+/-0.3; 2.5+/-0.3 and 3.6+/-0.3; 2.9+/-0.3 and 2.7+/-0.3, respectively. The average total number of viable embryos was greater (P=0.01) in supplemented cows (3.5+/-1.1; 5.4+/-1.4 and 7.5+/-1.2 in control, T800 and T1200, respectively), but less (P=0.01) in heifers (7.5+/-1.2; 5.6+/-1.2 and 4.0+/-1.1 in control, T800 and T1200, respectively). Supplementation injections of beta-carotene associated to tocopherol improved embryo quality in superovulated Holstein cows, in the present experimental conditions and may be advantageous in similar embryo production systems. However, at dosages applied in the present experiment, this treatment should not be recommended for nulliparous heifers.     

Embryo production from superovulated cattle following insemination of sexed sperm

Two trials were conducted to ascertain fertilization rate, embryo quality and numbers of transferable embryos in superovulated heifers and cows inseminated with sexed sperm. Inseminates contained 2 x 10(6), 10 x 10(6) or 20 x 10(6) total sperm enriched for the X- or Y-chromosome ( approximately 90%) by flow cytometry/cell sorting. Non-sexed inseminates contained 40 x 10(6) total sperm. Donors in each trial were allocated to one of each of the bulls included in that study. Each donor was inseminated with frozen/thawed sperm from the same bull for each treatment in successive courses of superstimulation with twice daily i.m. injections of FSH for 4 d. Heifers and cows were inseminated 12 and 24 h after visually observed standing estrus in Trial 1. In Trial 2, a single timed inseminate was used 70-72 h following PGF(2alpha). Ova/embryos were collected non-surgically 7-7.5 d after insemination. In both trials, fewer ova were fertilized with sexed versus non-sexed treatments and with 2 x 10(6) sexed sperm compared to higher doses (P < 0.05). However, insemination of 20 x 10(6) total sexed sperm of >or=90% purity resulted in similar numbers of transferable embryos of the desired sex compared to that for non-sexed sperm.     

Estimation and manipulation of glutathione levels in prepuberal mouse ovaries and ova:Relevance to sperm nucleus transformation in the fertilized egg

Glutathione (GSH), the major low-molecular-weight thiol in mammalian cells, is believed to be a necessary factor for the transformation of the disulfide-stabilized sperm nucleus into the male pronucleus after fertilization. Its concentration in mouse ova, isolated from the ampulla of the oviduct after hormone-induced superovulation of 3–4-week-old mice, has been determined by an enzymic cycling microassay. The level found was 1.80 pmol per ovum. Mean ovum diameter was estimated as 71–72 μm, indicating a GSH concentration of 9–10 mM in the mouse egg. Administration of L-buthionine S, R-sulfoximine, an inhibitor of GSH biosynthesis, during the 2 days preceding ovulation, reduced ovum GSH content below 0.20 pmol (<1.0 mM). The mean GSH concentration of the hormone-stimulated ovaries was reduced from 3.2 mM to 0.2 mM under these conditions. It has also been demonstrated that measurement and manipulation of ovum and ovarian levels of GSH can aid in studying its function in ovaries, ova, and early embryos. Hormone-induced superovulation was achieved in BSO-treated prepuberal C57B1/6 X SJL mice whose ovaries contained less than 10% of control levels of GSH. Over 50% of the isolated ova were fertilized in vitro. However, abnormal one-cell embryos resulted in which the maternally derived pronucleus coexisted with an unchanged sperm nucleus, thus confirming that adequate levels of GSH are necessary for initiating transformation of the fertilizing sperm nucleus.     

The effect of estradiol valerate on follicular dynamics and superovulatory response in cows with Syncro-Mate-B implants

Two experiments were designed to evaluate the effect of estradiol valerate on follicular dynamics and superovulatory response in cows with Syncro-Mate-B (SMB)implants. In Experiment 1, 5 mg estradiol valerate (E(2)), injected at the same time as superstimulation treatments were initiated, resulted in fewer corpora lutea (CL), ova/embryos collected and fertilized ova (P<0.05) than if E(2) was administered with the SMB implant 7 days earlier. In Experiment 2, 31 beef cows and 26 Holstein cows were placed in one of four treatment groups. Group I (control) cows were superstimulated on Day 9 (estrus=Day 0). On Day 2, cows in Groups II, III, and IV received SMB and cows in Group III received E(2). On Day 9, cows in Group IV received E(2), and all cows were superstimulated with Folltropin. The number of CL did not differ (P>0.19) among groups. However, there were more follicles < 10 mm and fewer fertilized ova and transferable embryos (P<0.02) in Group IV cows. Ovarian ultrasonography revealed that the diameter of the largest follicle in Group III cows declined from Day 2 to Day 7 and subsequently increased until Day 13. In contrast, Groups I, II and IV were characterized by apparently linear growth between Days 2 and 13. Differences (P<0.05) were detected between Days 5 and 9. Mean diameter of the largest follicle was smaller for cows in Group III than for the remaining groups on Day 9. It was concluded that SMB did not adversely affect superovulatory response and that E(2) administration resulted in atresia of the antral follicles in the cows with SMB implants.     

Factors affecting success of embryo collection and transfer in large dairy herds

Our objective was to evaluate factors that affected the success of embryo transfer programs in large dairy herds. Non-lactating donor cows produced a larger number of ova/embryos (P<0.01) and viable embryos (P<0.01) than lactating cows. The interaction between season and donor class was correlated with the proportion of ova/embryos classified as fertilized (P=0.03), because lactating donors had fewer fertilized ova in the summer. There was no correlation between 305-day mature equivalent milk yield and response to superstimulation. Although the interval between superstimulation protocols was correlated with the number of ova/embryos (P=0.03), there was no correlation with the number of viable embryos. Pregnancy per embryo transfer (P/ET) in heifer recipients was correlated with embryo quality grade (P<0.01), season (P=0.04), and whether embryos were fresh or frozen/thawed (P<0.01). Lactating recipient cows tended to have a lower rate of P/ET during the summer (P=0.12 to P=0.08). Synchronization protocols tended to be (P=0.06; Herd 1) or were (P=0.02; Herd 2) correlated with P/ET. Lactating cows receiving vitrified IVF embryos had a lower (P=0.01) P/ET than those receiving fresh IVF embryos, especially in the summer (P=0.09). Milk yield was not correlated with P/ET. The use of heat abatement systems is critical to improve embryo production and P/ET. Synchronization protocols that optimized synchrony of ovulation may increase fertility of recipient cows and eliminate the need for estrous detection.     

Superovulatory responses of Holstein cows

Approximately 1000 registered cows and heifers were superovulated one to 10 times. Nonsurgical embryo recoveries were performed on all donors which exhibited estrus. Healthy donors produced more total ova and cleaving embryos and had a higher ovum recovery rate, fertilization rate and pregnancy rate from embryos transferred than did cows classified as infertile. While ovum number was not affected during 10 repeated superovulations, fertilization rate and embryo number decreased. The number of ova recovered from healthy cows was affected by season, and from infertile cows by the day of the estrous cycle on which FSH was started and by the number of days since calving. More ova were recovered from infertile cows synchronized with prostaglandins prior to superovulation than following a natural estrous cycle. The number of embryos recovered from infertile cows was affected by age and from healthy cows by daily milk production. Fertilization rates in both healthy and infertile cows were affected by age, time since calving, daily milk production, day of cycle FSH was injected and season. There was no effect of the day of recovery on the number of ova or embryos recovered from healthy or infertile cows.     

Effect of dominant follicle removal before superstimulation on follicular growth, ovulation and embryo production in Holstein cows

This study was to investigate whether removing the dominant follicle 48 h before superstimulation influences follicular growth, ovulation and embryo production in Holstein cows. After synchronization, ovaries were scanned to assess the presence of a dominant follicle by ultrasonography with a real-time linear scanning ultrasound system on Days 4, 6 and 8 of the estrus cycle (Day 0 = day of estrus). Twenty-six Holstein cows with a dominant follicle were divided into 2 groups in which the dominant follicle was either removed (DFR group, n=13) by ultrasound-guided follicular aspiration or left intact (control group, n=13) on Day 8 of the estrus cycle. Superovulation treatment was initiated on Day 10. All donors were superovulated with injections of porcine FSH (Folltropin) twice daily with constant doses (total: 400 mg) over 4 d. On the 6th and 7th injections of Folltropin, 30 mg and 15 mg of PGF2alpha (Lutalyse) were given. Donors were inseminated twice at 12 h and 24 h after the onset of estrus. Embryos were recovered on Day 6 or 7 after AI. During superstimulation, the number of follicles 2 to 5 mm (small), 6 to 9 mm (medium) and > or = 10 mm (large) was determined by ultrasonography on a daily basis. At embryo recovery, the number of corpora lutea (CL) was also determined by ultrasonography and blood samples were collected for analysis of progesterone concentration. Follicular growth during superstimulation was earlier in the DFR group than in the control group. The number of medium and large follicles was greater (P < 0.01) in the DFR group than in the control group on Days 1 to 2 and Days 3 to 4 of superstimulation, respectively. The numbers of CL (9.6+/-1.1 vs 6.1+/-0.9) and progesterone concentration (30.9+/-5.4 vs 18.6+/-3.5 ng/mL) were greater (P < 0.05) in the DFR group than in the control group, respectively. The numbers of total ova (7.7+/-1.3 vs 3.9+/-1.0) and transferable embryos (4.6+/-0.9 vs 2.3+/-0.8) were also greater (P < 0.05) in the DFR group than in the control group, respectively. It is concluded that the removal of the dominant follicle 48 h before superstimulation promoted follicular growth, and increased ovulation and embryo production in Holstein cows.     

Probable influence of ova and embryo prostaglandins in the differential ovum transport in pregnant and cycling rats.

In this study we explored the possible underlying mechanism(s) of the differential transport of unfertilized and fertilized ova in cycling and pregnant rats. The number of ova recovered from rat oviducts and uterus was not significantly different in estrus, metestrus and diestrus but dropped sharply at proestrus. When estrus rats were injected with indomethacin (10(-6)), a well known inhibitor of cyclooxygenase, delivered into both ovarian bursae, and sacrificed next day at metestrus, the number of ova in the oviduct was significantly smaller (p less than 0.025) than in controls at metestrus. On the other hand, when diestrus rats were injected with PGE1 (10(-6)) delivered into both ovarian bursae, and sacrificed next day at proestrus, no ova were found in the oviducts, and only a few of them were in the uterus. When fertilized ova were recovered from oviducts and uteri at day 4 of pregnancy (corresponding to proestrus of cycling rats) an average of 4 embryos were still found in the oviducts, proving a differential ovum transport between cycling and pregnant rats. In order to establish if there exists any ova or embryo releasing factor responsible for this difference, the prostaglandins released to the incubation medium by ovum or 3-day embryo were measured. Unfertilized ova produced significantly more PGE1 (p less than 0.05) than PGE2 or PGF2 alpha. The same pattern of PG production was observed with incubated embryos, but in this case the amount of PGE1 released was significantly higher (p less than 0.01) that the PGE1 released by unfertilized ova.(ABSTRACT TRUNCATED AT 250 WORDS)     

Repeatability of response to superovulation in Brangus cows

Repeatabilities of total numbers of ova (TO) and morphologically normal embryos (MNE) recovered following superovulation of Brangus cows were estimated from records of 210 attempts involving 75 cows. Repeatability of the number of TO recovered was 0.22 +/- 0.08 and the number of MNE recovered was 0.19 +/- 0.08. Overall mean numbers of TO and MNE were 9.0 and 5.0, respectively. The mean numbers of TO recovered were 8.9 from the first treatment, 9.5 from the second treatment, and 8.6 from the third treatment. The mean numbers of MNE recovered were 5.2 from the first treatment, 4.6 from the second treatment, and 5.2 from the third treatment. Regressions of TO or MNE numbers per collection indicated that the superovulatory response did not decline significantly with repeated superovulation.     

Asymmetry in the ovary and uterus of the golden hamster (Mesocricetus auratus)

On Day 9 of pregnancy (day of mating = Day 1), the number of corpora lutea in the right ovary was greater than that in the left (mean +/- s.e.m. 9.3 +/- 0.1 and 6.5 +/- 0.3 respectively; N = 70). Although the percentages of ova fertilized on the left and right side were not different (82% and 94%), the percentage wastage was higher on the left side (20%) than the right (14%). A significant difference in sperm numbers in the right (2.8 X 10(6] and the left (0.5 X 10(6] uterine horns were found 1.5 h after mating in 51 females. Morphometric measurements of the lower uterine luminal size showed that the right side (103.9 mm3) was larger than the left side (88.9 mm3; N = 5). It is obvious that there is structural and functional asymmetry in the ovary and uterus in the golden hamster.