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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.

Animals↗

Hormonal state affects recovery from frontal cortex lesions in adult female rats.

Pseudopregnant and normal cycling female Sprague-Dawley rats were trained on a delayed-spatial alternation task. After acquisition training, the subjects received frontal cortex lesions and were subsequently tested for retention. The pseudopregnant animals with lesions were less impaired than their normal cycling counterparts. Learning and retention testing were not affected by hormonal status in the noninjured controls. Histological examination showed that the normal cycling rats with frontal cortex lesions possessed enlarged ventricles, indicating the presence of edema in this surgical group. However, brain tissue of pseudopregnant subjects given the same lesion and sham operates of both hormonal states did not display this characteristic of edema. We speculate that the behavioral and anatomical impairment observed in the normal cycling females with brain injury is due to higher levels of vasopressin, a potent vasoconstrictor synchronized with brain injury in our manipulation.

Animals↗

Aggressive behavior in female hamsters: the hormonal basis for fluctuations in female aggressiveness correlated with estrous state.

The frequency and sequencing of aggressive behaviors by naive female hamsters change during series of brief encounters, probably because of the lack of stable dominance relations. Such initial encounters seem most representative of interactions likely in free-ranging hamsters and have been emphasized in studies of the hormonal mediation of female aggression. Nonestrous females exhibit intense aggression toward conspecifics of either sex. Estrous females are not aggressive and spend much time in lordosis, indicative of sexual receptivity. The inhibition of fighting on estrous day depends on estrogen and progesterone. Whereas oil-injected adrenalectomized-ovariectomized females fight at high levels, comparable with intact nonestrous females, the combination of 17 beta-estradiol benzoate and progeterone suppresses fighting completely. In contrast, replacement of estradiol, progesterone, or testosterone propionate individually has on consistent effect. Hypophysectomized females also fight at high levels, indicating that pituitary hormones are not required for vigorous aggression. Further, individual anterior pituitary hormones did not produce marked changes in fighting. These results emphasize the roles of estrogen and progesterone in synchronizing aggression with current reproductive state.

Adolescent↗

Adrenal gland involvement in synchronizing the preovulatory release of LH in rats.

In this study we have demonstrated that acute adrenalectomy (1000 hr proestrus) has no effect on the release of LH on proestrous afternoon. However, chronic adrenalectomy results in the loss of some factor responsible for synchronizing the preovulatory LH surge. Since this investigator has shown previously (15) that progesterone can influence the timing of LH release in ovariectomized and ovariectomized-adrenalectomized animals, it is most likely that adrenal progesterone is involved in synchronizing this event.

Adrenal Glands↗

Synchronization of oestus and timed insemination of mares.

Oestrus was synchronized in 116 mares by means of an i.m. injection of prostaglandin F-2 alpha (Day 0) and of fluprostenol (a PG analogue) on Day 16. Mares were then randomly divided into three groups. Group A mares (N = 30) were given 2500 i.u. hCG I.M. ON Day 20 and artificially inseminated on Day 21 without detection of oestrus. Group B mares (N = 32) were given 2500 i.u. hCG i.m. on Day 20 and inseminated on Days 21 and 23, also without oestrus detection. Group C mares (N = 54) were teased on Days 18, 19, 21, 23 and 25 and inseminated on Days 19, 21, 23 and 25 while they were in oestrus. Semen was collected by artificial vagina from 3 stallions. One-third of the mares in each group were assigned to each stallion at random. The gel-free fraction was divided equally among the mares, and used within 1 h of collection. Pregnancy rates at about 60 days of gestation were not significantly different. A high rate of synchronization of oestrus (80%) was attained within 48 h of treatment with fluprostenol.

Animals↗

Estrous response and pregnancy rates following calf removal in beef cows treated with prostaglandin F(2)alpha.

Five hundred fifty-four suckled beef cows in three herds were allotted within postpartum interval to one of four treatments. All cows received two injections of prostaglandin F(2)alpha (PGF(2)alpha) 11 days apart. Treatment I served as a control. Calves were removed for 48 hr following the first injection of PGF(2)alpha in treatment II. Calves were removed similarly after the second injection of PGF(2)alpha in treatment III and after both injections of PGF(2)alpha in treatment IV. Pregnancy rates at the synchronized service, by 24 days and by 45 days of breeding were not (P>0.05) affected by treatment. Similarly, the treatments had no significant (P>0.05) effect on percentage of animals exhibiting estrus following the first and second injections of PGF(2)alpha.

Journal Article↗

Intracellular free calcium in response to oxytocin in pig endometrial cells.

Intracellular free calcium concentration ([Ca2+]i) in response to oxytocin (OT) was studied in stromal, glandular epithelial and luminal epithelial cells obtained from the endometrium of gilts 16 days post-estrus. The amplitude of increased [Ca2+]i in response to 100 nM OT was greatest in stromal cells, intermediate in glandular epithelial cells and not evident in luminal epithelial cells. During continuous OT administration, stromal cells responded initially with a synchronous spike of [Ca2+]i that did not require extracellular Ca2+ and then displayed spontaneous asynchronous [Ca2+]i spikes that required extracellular Ca2+. Each cell possessed its own characteristic response. Increasing concentrations of OT induced an increasing percentage of stromal cells responding, with some cells having nearly equal [Ca2+]i responses at all concentrations and others having graded [Ca2+]i responses as the concentration of OT increased. These results are consistent with the proposed mechanism of OT action in pig endometrium involving activation of phosphoinositide-Ca2+ signaling pathway.

Animals↗

Interspecies implantation and mitochondria fate of panda-rabbit cloned embryos.

Somatic cell nuclei of giant pandas can dedifferentiate in enucleated rabbit ooplasm, and the reconstructed eggs can develop to blastocysts. In order to observe whether these interspecies cloned embryos can implant in the uterus of an animal other than the panda, we transferred approximately 2300 panda-rabbit cloned embryos into 100 synchronized rabbit recipients, and none became pregnant. In another approach, we cotransferred both panda-rabbit and cat-rabbit interspecies cloned embryos into the oviducts of 21 cat recipients. Fourteen recipients exhibited estrus within 35 days; five recipients exhibited estrus 43-48 days after embryo transfer; and the other two recipients died of pneumonia, one of which was found to be pregnant with six early fetuses when an autopsy was performed. Microsatellite DNA analysis of these early fetuses confirmed that two were from giant panda-rabbit cloned embryos. The results demonstrated that panda-rabbit cloned embryos can implant in the uterus of a third species, the domestic cat. By using mitochondrial-specific probes of panda and rabbit, we found that mitochondria from both panda somatic cells and rabbit ooplasm coexisted in early blastocysts, but mitochondria from rabbit ooplasm decreased, and those from panda donor cells dominated in early fetuses after implantation. Our results reveal that mitochondria from donor cells may substitute those from recipient oocytes in postimplanted, interspecies cloned embryos.

Abdominal Muscles↗

JSAR Innovative Technology Award. Development of ovulation synchronization and fixed time artificial insemination in dairy cows.

Recently, reproductive management has become more difficult as a result of increased herd size. Problems with missing estrous signs and decrease in conception rate by artificial insemination (AI) performed at wrong timing have caused low AI conception rates. In 1995, ovulation synchronization and fixed-time AI (Ovsynch/TAI) was developed in the USA as a new reproductive technology, which was accepted as an useful reproductive management tool in many countries. However, no information on the use of Ovsynch/TAI was available in Japan. It was, therefore, warranted to show the ovulation rate and conception rate after Ovsynch/TAI using gonadotropin releasing hormone analogue (GnRH-A, fertirelin acetate) and prostaglandin F2alpha (PGF2alpha)-THAM, both were commercially available in this country. The conception rate after Ovsynch/TAI has been known to vary among different herds and individuals. Investigation and analysis of factors affecting the conception rate was also warranted to improve the conception rate. A series of experiments were carried out to establish Ovsynch/TAI using domestically produced GnRH-A and PGF2alpha and to study factors affecting conception rate after Ovsynch protocol. Ovsynch using 100 microg GnRH-A and 25 mg PGF2alpha were observed using ultrasonography. As a result, a high synchronization rate of ovulation at 16 to 20 h after the second GnRH injection was confirmed. The conception rate after Ovsynch/TAI was compared in 87 cows with the conception rate after AI at estrus induced by PGF2alpha (139 cows). Conception rate after Ovsynch/TAI was higher than the figure after AI at induced estrus (59.1% vs 20.9%, P<0.05). The dose of GnRH-A was also studied and a practical dose of GnRH-A was found to be 50 microg per cow. To clarify some factors affecting the conception rate after Ovsynch/TAI, 1,558 cows were investigated for the state of their ovaries, days after calving, parity, season, ovarian cyclicity postpartum and nutritional state at the day of Ovsynch. The overall conception rate after Ovsynch/TAI was 51.5%. Fifty-six cows (3.6%) showed estrus at 6 to 7 d after the first injection of GnRH-A. The conception rate after Ovsynch/TAI was low in cows that were 40 to 60 d postpartum, those in their 5th lactation or more, those bred in July to August, and those recovering ovarian cyclicity later than 56 d postpartum. The conception rate after Ovsynch/TAI was high in cows in which body condition score (BCS) was 3.75 at dry period and 3.0 at the day of Ovsynch. In conclusion, Ovsynch/TAI is an effective tool for the reproductive management of dairy cows. A steady and sufficient conception rate after Ovsynch/TAI could be expected by taking the factors affecting the conception rate into the consideration.

Animals↗

Effects of dietary supplementation with omega-3 polyunsaturated fatty acids on some reproductive characteristics in gilts.

The main objective of this study was to determine the effects of dietary supplementation with omega-3 polyunsaturated fatty acids on ovulation rate and litter size at day 27 post-mating in gilts. Estrous cycles were synchronized in crossbred gilts (n=48) using a progestin (Matrix; Intervet America Inc., Millsboro, DE, USA), fed at a rate of 15 mg/day for 18 days. Following progestin withdrawal, gilts were checked for estrus twice daily and mated by artificial insemination. At 27 days post-mating, gilts were killed and reproductive tracts collected and examined. Beginning 17 days before the initiation of progestin therapy and continuing until slaughter, gilts were fed 2.27 kg of a basal diet (n=24) or the basal diet supplemented with 1% Fertilium (JBS United, Inc., Sheridan, IN) (n=24), a source of omega-3 polyunsaturated fatty acids. All gilts in the treatment and control groups were pregnant as evidenced after examination of dissected reproductive tracts at approximately 27 days post-mating. Ovulation rate (17.5 vs. 17.9; SEM=0.5; p=0.61), number of embryos (14.5 vs. 14.3; SEM=0.6; p=0.77), embryo weight (1.11 vs. 1.14 g; SEM=0.02; p=0.45), and crown-rump length (26.4 vs. 26.9 mm; SEM=0.27; p=0.28), were similar for control and Fertilium-fed gilts, respectively. Backfat thickness at day 27 post-mating was greater (p<0.01) for Fertilium-fed gilts (14.0+/-0.5 mm) compared with controls (12.3+/-0.5 mm). The present treatment with Fertilium for the period of approximately 61 days did not alter ovulation rate and litter size at day 27 post-mating in gilts.

Animal Feed↗

Clinical experience with embryos produced by handmade cloning: work in progress.

Nuclear transfer in cattle has been shown to cause a high frequency of conceptus loss, excessive accumulation of allantoic fluid, increased birth weight as well as peri- and neonatal deaths. The aims of this preliminary study were to investigate the in vivo development of embryos and fetuses produced by a novel somatic cell cloning method, denominated handmade cloning (HMC), and to characterize the premature calves delivered by Caesarian section. Twenty-five day 7 fresh embryos including seven blastocysts produced by aggregation of two day 4 embryos, and seven vitrified embryos were transferred to synchronized Holstein-Friesian heifers. Embryos produced by aggregation had higher in vivo developmental competence than single embryos (67% versus 38% pregnancy rate on day 28). On days 28, 42, 63 and 250 after estrus, 12 (48%), 5 (20%), 3 (12%) and 2 (8%) recipients of fresh embryos remained pregnant, while 1 recipient of a vitrified embryo was pregnant. One recipient was euthanized due to development of hydrallantois. Caesarian sections were performed on the remaining three recipients on days 252 or 259. The premature calves weighed 60 kg, 47 kg and 45 kg, respectively, and displayed increased weights of body, heart, liver, kidneys, thyroid glands and increased size of placentomes. Furthermore, they had reduced respiratory function, hypoxia, acidosis and altered glucose metabolism. In conclusion, these preliminary data show that handmade somatic cell cloning resulted in an overall delivery rate of 9%, one case of hydrallantois (3%), oversized placentomes and fetuses, disproportionate growth of several internal organs and metabolic immaturity of the premature calves.

Animals↗

Mounting behavior as affected by stage of estrous cycle in Holstein heifers.

Nine Holstein heifers whose estrous cycles were synchronized were observed during three subsequent consecutive cycles. Observations of heifers during each periestrus period began when the first heifer reached day 15 of an estrous cycle and ended 36 h after onset of estrus for the ninth heifer. There were 118 1-h observations at 6-h intervals during the three periestrus periods. Estrous cycles were divided into postestrual, luteal, preestrual, and estrual stages by blood progesterone profiles and estrus. Heifers of these four stages accounted for 27.5, 48.3, 15.4, and 8.8% of heifers available to mount during the 118 observations, and heifers of each stage made 9.0, 5.2, 18.9, and 66.9% of attempted mounts. When number of heifers in estrus simultaneously increased from one to five, total attempted mounts and stands per hour increased from 10.1 to 63.5 and from 4.5 to 48.5. Stands per heifer per hour increased significantly as number of heifers in estrus increased from one to two and tended to increase as number in estrus increased further. Presence of one preestrual heifer in a group containing one estrual heifer increased total stands per hour from 2.3 to 5.0. Efficiency of detection of estrus may depend on proportion of animals within estrous cycle stages that have highest mounting activity.

Animals↗

Superovulation treatments and embryo transfer in Angora goats.

A high incidence of early luteal regression after PMSG superovulation was associated with low recovery of embryos from reproductive tracts of Angora goats flushed later than Day 5 after onset of oestrus. Embryos were successfully recovered (mean 7.9/female) by flushing on Days 2-5. Mean ovulation rate after an FSH regimen (16.1 +/- 0.8) was significantly higher than that after a single injection of PMSG (10.8 +/- 1.2). Fertilization rate and survival of embryos following transfer to naturally synchronized recipient feral goats did not differ between the two gonadotrophin regimens: the mean number of kids born to 47 donors treated with FSH (7.5 +/- 0.6) was significantly greater than that to 28 donors treated with PMSG (4.8 +/- 0.6). Irrespective of hormonal treatment, the numbers of embryos recovered and of kids born were correlated with ovulation rate (r = 0.82, P less than 0.001 for both). Embryo survival was influenced by ovulation rate in recipients, with 52%, 63% and 75% of transferred embryos being carried to term by recipients with 1,2 and 3 CL, respectively (P less than 0.01). More embryos survived (65%) when 2 embryos were transferred to each recipient than when 1 (51%) or 3 (48%) were transferred. In recipients receiving 2 embryos, survival was significantly improved by transfer of both embryos to the same oviduct (70%) than when one was transferred to each oviduct (62%). The percentage survival of embryos was optimal when oestrus of recipients was synchronized within +/- 1 day of oestrus in donors.

Animals↗

Variation in the timing of the reproductive season among breeds of sheep in relation to differences in photoperiodic synchronization of an endogenous rhythm.

Photoperiod may regulate seasonal reproduction either by providing the primary driving force for the reproductive transitions or by synchronizing an endogenous reproductive rhythm. This study evaluated whether breed differences in timing of the reproductive seasons of Finnish Landrace (Finn) and Galway ewes are due to differences in photoperiodic drive of the reproductive transitions or to differences in photoperiodic synchronization of the endogenous rhythm of reproductive activity. The importance of decreasing photoperiod after the summer solstice in determining the onset and duration of the breeding season was tested by housing ewes from the summer solstice in either a simulated natural photoperiod or a fixed summer-solstice photoperiod (18 h light:6 h dark; summer-solstice hold). Onset of the breeding season within each breed did not differ between these photoperiodic treatments, but Galway ewes began and ended their breeding season earlier than Finn ewes. The duration of the breeding season was shorter in Galway ewes on summer-solstice hold than on simulated natural photoperiod; duration did not differ between photoperiodic treatments in Finn ewes. The requirement for increasing photoperiod after the winter solstice for initiation of anoestrus was tested by exposing ewes from the winter solstice to either a simulated natural photoperiod or a winter-solstice hold photoperiod (8.5 h light:15.5 h dark). Onset of anoestrus within each breed did not differ between these photoperiodic treatments, but the time of this transition differed between breeds. These observations suggest that genetic differences in timing of the breeding season in Galway and Finn ewes do not reflect differences in the extent to which photoperiod drives the reproductive transitions, because neither breed requires shortening days to enter the breeding season or lengthening days to end it at appropriate times. These findings are consistent with the hypothesis that photoperiod synchronizes an endogenous rhythm of reproductive activity in both breeds and that genetic differences in timing of the breeding season reflect differences in photoperiodic synchronization of this rhythm.

Anestrus↗