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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↗

Effect of presence of clinical and subclinical endometritis at the initiation of Presynch-Ovsynch program on the first service pregnancy in dairy cows.

The present study examined the effect of presence of clinical or subclinical endometritis at the initiation of Presynch-Ovsynch estrous synchronization program on the first service pregnancy rate in dairy cows. Lactating Holstein cows (N=275) were given a thorough reproductive examination at 32-38 days in milk, 3 days prior to the scheduled start of Presynch-Ovsynch program. Based on the reproductive exam findings the cows were diagnosed and classified into three groups as clinical endometritis, subclinical endometritis and normal. All cows received two set-up injections of 25mg PGF(2alpha) (Lutalyse((R)), Pfizer Animal Health, New York, NY, USA) i.m., 14 days apart starting at 35-42 days in milk (DIM). All cows received 75microg of GnRH (Cystorelin, Merial, Iselin, NJ, USA) i.m. 14 days after the second pre-synchronization injection of PGF(2alpha), followed by a third injection of 25mg PGF(2alpha) i.m. 7 days later. Cows received a second injection of 75microg of GnRH i.m. 54h after the third PGF(2alpha), and received timed artificial insemination at the time of the second GnRH injection or 24h later. Multivariate logistic regression was used to analyze the odds of pregnancy at the first service. Variables included in the model were endometritis status (clinical endometritis, subclinical endometritis and normal), farm (two), presence of corpus luteum (CL, yes or no), timing of second GnRH in relation to AI (0 or 24h), sire fertility (bulls with greater compared with lesser estimated relative conception rates), parity (primiparous and multiparous) and their interactions. Of all variable included in the model, cows with corpus letuem (OR=1.83 versus OR=1.00; P=0.05) 3 days prior to the scheduled start of Presynch-Ovsynch program and primiparous cows (OR=1.00 versus OR=0.55; P=0.04) had increased odds of becoming pregnant at the first service. No differences were found in the odds of first service pregnancy among clinical, subclinical endometritis and normal cows (P>0.1). In summary, presence of clinical or subclinical endometritis at the initiation of Presynch-Ovsynch estrous synchronization program does not harm the first service pregnancy rate in dairy cows.

Animals↗

Synchronization of oestrus in adult female rats by utilizing the paradoxical effects of an LH-RH agonist.

Implantation of 25 microgram per rat of an LH-RH agonist, [Des-Gly10,D-Ala6,lProNHEt9]LH-RH, contained within a cholesterol matrix pellet caused a complete and continued suppression of oestrus in rats within 3 days. Removal of the pellets after 7 days led to synchronized oestrus 4 days later in 90-95% of the animals (N = 30); the remaining animals became oestrous on the 5th day. Fertility of these treated rats mated at the synchronized oestrus was equivalent to that of a control, sham-operated population allowed to mate over a 3-day period.

Animals↗

Comparison of the effectiveness of ovulation synchronization protocol in anestrous and cycling beef cows.

Applicability of ovulation synchronization protocol using GnRH and PGF(2alpha) (PGF) injection to anestrous beef cows remains controversial. We compared the effectiveness of the protocol in the anestrous stage of the beef cow with that in the cycling stage using the same animals. Ovaries of five Japanese Black and three Japanese Shorthorn cows were ultrasonographically examined, and blood samples were collected daily for hormonal analyses. Each animal received the protocol twice (Day -6 to -8: GnRH, Day 0: PGF, Day 2: GnRH). Additional blood samples were taken before and after GnRH injection for LH and FSH measurements to evaluate the pituitary function. For the ovarian status at the onset of the protocol cows were divided into anestrous (n=8) and cycling (n=8) stages. There was no significant difference in size of the dominant follicle at the first and second GnRH injections, and in the magnitude of the pituitary response to GnRH between the two stages. However, the size of the corpus luteum and progesterone concentrations at the PGF injection in the anestrous stage were significantly smaller and lower (P<0.01), respectively, and ovulation synchronization rate in the anestrous stage was significantly lower (P<0.05) than in the cycling stage. In conclusion, ovulation synchronization protocol in anestrous beef cows has limited effectiveness.

Animals↗

Male influence on oestrous cycles in female woolly opossum (Caluromys philander).

Plasma progesterone concentrations and the occurrence of oestrous cycles were studied in isolated woolly opossums subsequently subjected to male influences during a 40-day period. Pairing (N = 48) or exposure to male urine (N = 15) resulted in all females exhibiting oestrous during the stimulation phase, providing evidence that the activation of ovarian activity in the woolly opossum involves pheromonal cues from males. The latency of occurrence of oestrous in stimulated females depended upon their sexual state before male stimulation. In anoestrous females, the mean latency was 20.7 +/- 0.9 days (N = 35), a value which agrees with the duration of the follicular phase. In females which first entered oestrous before male stimulation, the latency of induced oestrous was inversely correlated to the date of occurrence of the previous oestrous. The inter-oestrous interval was normal (38.1 +/- 1 days, N = 5) when females were in oestrous at the beginning of male stimulation. In contrast, the inter-oestrous interval was significantly shortened (28.7 +/- 2 days, N = 7) or lengthened (51.1 +/- 1.7 days, N = 16) depending on whether females were in the luteal or follicular phases at the beginning of male stimulation. During pairing several females became pregnant and gave birth 24 +/- 0.9 days (N = 13) after copulation. In the woolly opossum, the response to male influences involves mechanisms similar to those observed in eutherians and results in enhancement and synchronization of oestrous cycles in females. Pheromonal interactions could play an important role in synchronizing oestrous cycles in wild females during the dry season, a period when animals regroup to feed on spatially localized food resources.

Animals↗

Administration of 6-methoxybenzoxazolinone (MBOA) does not augment ovulatory responses in St. Croix White ewes superovulated with PMSG.

The objective of this investigation was to examine the effects of 6-methoxy-benzoxazolinone (MBOA), a plant compound that resembles melatonin and alters ovarian function in rodents, in combination with PMSG on superovulatory responses in the cycling ewe. In Experiment I, St. Croix White ewes (n = 44) were synchronized (intra-vaginal progestin sponge) for 14days followed by hCG (750 IU) at 1 day after sponge removal (day 0). Ewes were assigned to one of six treatments administered on day -1: Control (no PMSG or MBOA; n = 7); PMSG (1000 IU i.m.; n = 7); Low MBOA (0.43 mg/kg i.m.; n = 7); High MBOA (1.15 mg/kg i.m.; n = 7); Low MBOA + PMSG (n = 8); High MBOA + PMSG (n = 8). In Experiment II, St. Croix White ewes (n = 24) were synchronized (progestin CIDR) for 14 days followed by hCG on day 1 after CIDR removal (day 0). Ewes were assigned to one of three treatments administered on day -1: Control (n = 8); PMSG (n = 8); Low MBOA+PMSG (n = 8). Laparoscopy was performed on day 9 to assess numbers of corpora lutea (CL) and visible follicles on each ovary. Blood samples were collected on day -13, -1, 0, 1, and days 6 or 7-12 for analysis of serum progesterone (P4) by RIA. Treatment groups receiving PMSG (alone or with MBOA) exhibited greater (P < 0.05) serum concentrations of P4 post-synchrony than Control and MBOA-only groups. Ovulation rate was lower (P < 0.05) for Control and MBOA-only treated ewes than ewes receiving PMSG. Ovulation rate in ewes treated with MBOA alone was similar (P > 0.10) to Controls, and PMSG treatment alone did not differ (P > 0.10) from MBOA + PMSG treatment. Ewes treated with PMSG alone did not differ (P > 0.10) in follicle number from High MBOA + PMSG treated ewes, however, Low MBOA + PMSG treated ewes had greater numbers of follicles at day 9 (P < 0.05) than the PMSG or High MBOA + PMSG groups in Experiment I; although, this was not replicated in Experiment II with numbers of follicles in the Low MBOA + PMSG group being similar (P > 0.10) to PMSG alone. In summary, the addition of MBOA in combination with PMSG as part of a synchronization-superovuation protocol in the ewe did not increase ovulation rate.

Animals↗

Responsiveness to progestagen-eCG-cloprostenol treatment in goat food restricted for long period and refed.

For 6 months, 10 adult Saanen crossbred goats were fed undernutrition diet (70% maintenance), and finally five goats were refed for 6 weeks with 150% maintenance. In all animals oestrus was synchronized using 45 mg FGA vaginal sponge for 11 days, 300 IU eCG and 50 microg cloprostenol 48 h prior to sponge removal. From oestrus onset, during a 24-h period, blood samples were collected for oestradiol and NEFA assay. Ovulation was verified by laparoscopy 3 days after sponge removal. Body mass loss was 18.62 +/- 3.03% of initial weight and in refed goats body weight recovery was 90.63 +/- 3.56%. NEFA level was higher in restricted goats (p < 0.05). Fifty per cent of underfed goats (2/4) and all refed goats (4/4) exhibited oestrus and ovulation. Significant relationship (p < 0.05) was found between weight loss and the interval sponge removal-oestrus onset (r = 0.91) or ovulation rate (r = 0.70). Only in the refed group was the ovulation rate related to the oestradiol amount (r = 0.99) (p < 0.05). Collectively results showed that a short period of improved feeding re-established the responsiveness of oestrus synchronization in chronically fasted goats.

Animal Feed↗

Interleukin 8 in the cervix of non-pregnant ewes.

Studies of cervical artificial insemination of ewes at hormone-synchronized oestrus indicate that the cervix remains relatively impenetrable to semen, in contrast to naturally breeding animals. During parturition the inflammatory response plays an important part in cervical dilation and possibly, to a lesser extent, in the non-pregnant cervix at oestrus to facilitate the transcervical transport of semen. The expression of interleukin 8 (IL-8), a pro-inflammatory cytokine in the ovine cervix, has been mapped and quantified, using semi-quantitative in situ hybridization, to ascertain the role played by inflammation in the ovine cervix during natural and artificially induced oestrous cycles. IL-8 gene expression was observed in both the luminal epithelium and fibroblastic cells of the cervix. The presence of IL-8 was confirmed using immunohistochemistry. IL-8 gene expression in the luminal epithelium varied throughout the oestrous cycle and was highest at oestrus and at day 5 of the oestrous cycle. In ewes artificially induced to ovulate, either by the withdrawal of progesterone pessaries after treatment for 12 days, or by two i.m. injections of prostaglandin 9 days apart, IL-8 gene expression at oestrus was significantly lower than it was at natural oestrus. Insemination increased IL-8 gene expression in progesterone-synchronized ewes. These data support the hypothesis that IL-8-induced inflammation is important in normal cervical function and that this process is inhibited during artificial synchronization of the oestrous cycle and is increased by exposure to semen.

Animals↗

Ovulation, fertilization and pronucleus development in superovulated gilts.

Estrus was synchronized in 45 gilts by ingestion of Zinc-Methallibur in the feed for 15 d. On Day 16 each gilts was treated with PMSG (1200 IU i.m.) followed in 72 h by hCG (500 IU i.m.). Gilts were inseminated 24 and 36 h after the onset of estrus followed by slaughter of groups (n = 4 or 5) at 40 h, 44 h, 48 h, 52 h, 56 h, 60 h and 64 h after hCG injection. Ovaries were evaluated macroscopically and oocytes/embryos were recovered by flushing the oviducts. The ovulation rate increased from 38% to 87% from 40 to 45 h and remained constant thereafter. At 40 h, 36% of oocytes were penetrated by a single spermatozoon. The rate of fertilization increased from 36% (40 h) to 59% (44 h), to 65% (48 h), to 73% (52 h), to 76% (56 h), 80% (60 h) and to 64% (64 h). At 40 h all fertilized ova contained a decondensed sperm head. After another 4 to 8 h early pronuclei were common, and 52 h after hCG treatment opposed pronuclei were predominant. The first cleavages were recorded 64 h after hCG injection.

Journal Article↗

Recent developments in oestrous synchronization of postpartum dairy cows with and without ovarian disorders.

This report reviews the most recent developments in prostaglandin-based oestrous synchronization programmes for postpartum dairy cows and addresses the efficiency of controlled breeding protocols based on such developments for cows with abnormal ovarian conditions. A double prostaglandin protocol applied 11-14 days apart seems to be capable of bringing most cows to oestrus. Because of varying oestrus onset times, improved conception rates are obtained following artificial insemination (AI) at detected oestrus rather than fixed-time AI in prostaglandin-treated cows. The administration of oestradiol or human chorionic gonadotrophin, or both these hormones, after prostaglandin treatment, improves the synchrony of oestrus yet does not enhance the conception rate. Progesterone-based treatments for oestrous synchronization are considered the most appropriate for non-cyclic or anoestrous postpartum dairy cows; prostaglandin alone being ineffective because of the absence of a mature corpus luteum in these cows. Improved oestrus synchrony and fertility rate have been reported using short-term progesterone treatment regimes (7-9 days) with or without oestradiol benzoate combined with the use of a luteolytic agent given 1 day before, or at the time of, progesterone withdrawal. The ovulation synchronization (Ovsynch) protocol, based on the use of gonadotrophin releasing hormone and prostaglandin, was developed to coordinate follicular recruitment, CL regression and the time of ovulation. This protocol allows fixed time insemination and has proved effective in improving reproductive management in postpartum dairy cows. However, timed AI following Ovsynch seems to have no beneficial effects in heifers, because of an inconsistent follicle wave pattern, and in anoestrous cows, given their lack of prostaglandin responsive CL. To date, there are several prostaglandin based, fixed-time insemination oestrous synchronization protocols for use in early postpartum dairy cows with ovarian disorders such as ovarian cysts and acyclicity.

Animals↗

Effect of gonadotropin-releasing hormone at estrus on subsequent luteal function and fertility in lactating Holsteins during heat stress.

The experiment used lactating Holstein cows (n = 94) from three herds in Mississippi. The experiment was conducted during late summer when temperatures were hot enough to cause a measurable stress response in cows in order to determine the effect of GnRH administration during estrus on fertility and subsequent luteal function of cows under these conditions. The mean ranges for ambient temperature, relative humidity, and temperature-humidity index during the study were 21.4 to 32.8 degrees C, 67 to 95%, and 21.6 to 29.6, respectively. After injection of PGF2 alpha for synchronization of estrus, cows were alternately injected with 100 micrograms of either GnRH or saline (control) at detection of estrus, followed by AI 10 to 12 h later. From 14 treated cows and 14 control cows, blood samples were drawn by venipuncture just prior to treatment and at 5-d intervals until 30 d after treatment. Serum progesterone concentrations were determined by radioimmunoassay. Mean progesterone concentrations were higher for the cows treated with GnRH than for the controls. The pregnancy rate from first AI was 28.6% for all treated cows and 17.7% for control cows. On d 20, 42.8% of the treated cows and 57.1% of the control cows exhibited progesterone concentrations that were typical of pregnancy. When pregnancy was diagnosed in these cows after d 45, pregnancy rates had decreased significantly for controls but not for cows given GnRH at estrus, suggesting greater embryonic survival in treated cows. We concluded that GnRH treatment enhanced secretion of luteal progesterone and embryo survival.

Animals↗

Follicular development and reproductive endocrinology during a synchronized estrous cycle in heifers and mature cows displaying contrasting superovulatory responses.

Ovarian follicular development and plasma concentrations of progesterone (P4), estradiol-17 beta (E2), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were compared during a synchronized estrous cycle between heifers and mature cows displaying contrasting superovulatory responses. Six heifers < 2 years old with a history of good responses to superovulatory (SOV) treatment and six cows 9 to 13 years old with poor responses to SOV treatments were used. Follicular development was monitored by daily ultrasonography. Blood samples were collected two to three times daily for P4 and E2 and thrice daily for LH and FSH analysis. Intensive sampling (samples every 15 min for 6 hr) was performed at critical periods of follicular development to analyze the pulsatile secretion of gonadotropins. In both cattle groups, a transient increase (P = 0.0001) in E2 occurred 4 to 5.7 d after the preovulatory LH surge or 2.3 d before the dominant follicle reached its maximum size. FSH concentrations increased (P = 0.006) before the emergence of the second cohort of follicles and then decreased despite no change in the concentration of E2. Contrary to our expectation and despite differences between groups in terms of age, number of previous SOV treatments, and divergent responses to superovulation, follicular development was similar in both groups. However, during the luteal phase, concentrations of E2 and FSH and LH pulse amplitudes were less (P < or = 0.05) in cows than in heifers. Therefore, follicular development monitored by ultrasonography and endocrine profiles during a synchronized estrous cycle are of limited value to predict quality of embryo donors.

Aging↗

Effect of timing of prostaglandin administration, controlled internal drug release removal and gonadotropin releasing hormone administration on pregnancy rate in fixed-time AI protocols in crossbred Angus cows.

Two experiments were conducted to investigate the effects of timing of prostaglandin F2(alpha) (PGF2(alpha)) administration, controlled internal drug release device (CIDR) removal and second gonodotropin releasing hormone (GnRH) administration on the pregnancy outcome in CIDR-based synchronization protocols. In Experiment 1, suckled Angus crossbred beef cows (n = 580) were given 100 microg of GnRH+a CIDR on Day 0. Cows in Group 1 (modified Ovsynch-P) received 25 mg of dinoprost (PGF2(alpha)) and CIDR device removal on Day 8 (AM), 100 microg of GnRH 36 h later on Day 9 (p.m.), and fixed-time AI (FTAI) 16 h later on Day 10 (47.5+/-1.1 h after PGF2(alpha)). Cows in Group 2 (Ovsynch-P) received 25mg of PGF2(alpha) and CIDR device removal on Day 7 (p.m.), 100 microg of GnRH 48 h later on Day 9 and FTAI 16 h later on Day 10 (66.6+/-1.2 h after PGF2(alpha)). Pregnancy rates were 56.5% (170/301) for Group 1 and 55.6% (155/279) for Group 2, respectively (P = 0.47). In Experiment 2, beef cows (n=734) were synchronized with 100 microg of GnRH+CIDR on Day 0, 25 mg of PGF2(alpha) and CIDR device removal on Day 7 and either 100 microg of GnRH 48 h later on Day 9 (Ovsynch-P) and FTAI 16 h later on Day 10 (64.9+/-3.3 h from PGF2(alpha)) or 100 microg of GnRH on Day 10 (CO-Synch-P) at the time of AI (63.2+/-4.2 h from PGF2(alpha)). Pregnancy rates were 48.8% (180/369) for Ovsynch-P and 44.7% (163/365) for CO-synch-P groups, respectively (P = 0.11). In both experiments, there was a locationxtreatment interaction (P<0.05); pregnancy rates between locations were different (P < 0.05) in the Ovsynch-P group. In conclusion, in a CIDR-based Ovsynch synchronization protocol, delaying administration of prostaglandin and CIDR removal by 12 h, or timing of the second GnRH by 16 h, did not affect pregnancy rates to FTAI. Therefore, there may be an opportunity to make changes in synchronization protocols with out adversely affecting FTAI pregnancy rates.

Animals↗

Ovarian follicular wave synchronization and pregnancy rate after fixed-time natural mating in llamas.

The study was designed to compare the efficacy of treatments intended to induce follicular wave synchronization among llamas (Experiment 1), and to determine the effect of these treatments on pregnancy rates after fixed-time natural mating (Experiment 2). In Experiment 1, llamas were treated with: (1) saline (control, n=20); (2) estradiol and progesterone (E/P, n=20); (3) LH (LH, n=20); or (4) transvaginal ultrasound-guided follicle ablation (FA, n=20). The ovarian response was monitored daily by transrectal ultrasonography. The intervals from treatment to follicular wave emergence and to the day on which the new dominant follicle reached >/=7 mm, respectively, did not differ between the LH (2.1+/-0.3 days and 5.2+/-0.5 days, respectively) and FA groups (2.3+/-0.3 days and 5.0+/-0.5 days), but both were shorter (P<0.05) and less variable (P<0.01) than in the control group (5.5+/-1.0 days and 8.4+/-2.0 days), while the E/P group (4.5+/-0.8 days and 7.7+/-0.5 days) was intermediate. In Experiment 2, llamas at unknown stages of follicular development were assigned randomly to control, E/P, and LH groups (n=30 per group). A single, fixed-time natural mating was permitted 10-12 days after treatment. Ovulation rates did not differ among groups (control, 93%; E/P, 90%; LH, 90%; P=0.99), but the pregnancy rate was higher (P<0.05) for synchronized llamas (LH and E/P groups combined, 41/54) than for non-synchronized llamas (control group, 15/28). In conclusion, LH and FA treatments were most effective for inducing follicular wave synchronization, while E/P treatment was intermediate. Synchronization treatments did not influence ovulation rate subsequent to fixed-time natural mating, but a higher pregnancy rate in synchronized than non-synchronized llamas warrants critical evaluation of the effects of follicular status on the developmental competence of the contained oocyte.

Animals↗

Evaluation of two hormonal protocols for synchronization of ovulation and timed artificial insemination in dairy cows managed in grazing-based dairies.

To evaluate the efficacy of two hormonal protocols for synchronization of ovulation and timed artificial insemination (TAI) in dairy cows managed in grazing-based dairies, lactating dairy cows (n = 142) from two grazing-based dairies were randomly assigned to one of three treatment groups. Cows in the first group (Ovsynch) received 50 microg of GnRH (d -10); 25 mg of PGF2alpha (d -3), and 50 microg of GnRH (d -1) followed by timed AI on d 0. Cows in the second group (PGF + Ovsynch) received a modified Ovsynch and timed AI similar to Ovsynch but with the addition of 25 mg of PGF2alpha 12 d (d -22) before initiation of Ovsynch. Cows in the third group (control) received standard reproductive management in place on each farm. Luteolysis occurred in 90.5% of cows exhibiting luteal function on d -22 in the PGF + Ovsynch treatment group, whereas none of the cows in the Ovsynch group underwent luteolysis on d -22. Synchronization rate (i.e., ovulatory response at 48 h after the second GnRH injection), conception rates at TAI and pregnancy rates after 35 d of breeding were similar for cows in the Ovsynch and PGF + Ovsynch groups. The proportion of anovular cows at the first GnRH injection of the synchronization protocols (d -10) was similar for cows receiving Ovsynch (28.0%) and PGF + Ovsynch (30.7%), and conception rate at TAI was similar for cycling (45.8%) and anovular (30.0%) cows. The cumulative pregnancy rate was greater for cows receiving TAI compared with control cows after 7 d of breeding (41.2 vs. 20.0%) but did not differ at 35 d of breeding (54.9 vs. 60.0%). Administration of PGF2alpha 12 d before initiation of Ovsynch did not improve synchronization, conception, or pregnancy rate compared with the standard Ovsynch protocol. Synchronization of ovulation to initiate timed AI at the onset of the breeding season resulted in earlier establishment of pregnancy compared with standard reproductive management.

Animals↗