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A comparison of progestin-based protocols to synchronize ovulation and facilitate fixed-time artificial insemination in postpartum beef cows.

The experimental objective was to compare pregnancy rates after fixed-time AI in postpartum suckled beef cows following administration of two progestin-based protocols to synchronize ovulation. Cows (n = 424) at three locations (n = 208, 122, and 92 per location) were stratified by age, BCS, and days postpartum (DPP) and assigned randomly to one of the two treatment protocols. The MGA Select-treated cows (MGA Select; n = 213) were fed melengestrol acetate (MGA, 0.5 mg x cow(-1) x d(-1)) for 14 d and carrier for 8 d, and then GnRH (100 microg i.m. Cystorelin; d 26) was injected 12 d after MGA withdrawal, and PG (25 mg i.m. Lutalyse) was administered 7 d after GnRH. Cows assigned to the 7-11 Synch protocol (7-11 Synch; n = 209) were fed carrier for 15 d and MGA for 7 d, and then injected with PG on d 22 (d 7 of MGA), GnRH on d 26, and PG again on d 33. Artificial insemination was performed at fixed times for cows in both treatments at 60 or 72 h after d 33 PG for 7-11 Synch and MGA Select groups, respectively. All cows were injected with GnRH (100 microg of i.m. Cystorelin) at AI. There was no treatment x location interaction for age (P = 0.90), BCS (P = 0.64), or DPP (P = 0.93), and the results were therefore pooled for the respective treatments (age [7-11 Synch, 5.5 +/- 0.2; MGA Select, 5.5 +/- 0.2], BCS [7-11 Synch, 5.7 +/- 0.1; MGA Select, 5.6 +/- 0.1], and DPP [7-11 Synch, 41.1 +/- 1.1; MGA Select, 42.1 +/- 1.1]). Blood samples were collected 8 and 1 d before MGA or carrier to determine pretreatment estrous cyclicity (progesterone >or=1 ng/mL; 7-11 Synch, 59/209 [28%]; MGA Select, 54/213 [25%]; P = 0.50) and again on d 33 PG to evaluate treatment response as a percentage of cows with progesterone concentrations in serum >or=1ng/mL (7-11 Synch, 184/209 [88%]; MGA Select, 177/213 [83%]; P = 0.15). Pregnancy rates resulting from fixed-time AI did not differ (P = 0.25) between treatments (7-11 Synch, 128/209 [61%]; MGA Select, 142/213 [67%]), nor did pregnancy rates (P = 0.77) at the end of the breeding season (7-11 Synch, 198/208 [95%]; MGA Select, 204/213 [96%]). These data indicate that pregnancy rates were comparable after fixed-time AI, following administration of the 7-11 Synch and MGA Select protocols. Both protocols provide opportunities for beef producers to use AI and eliminate the need to detect estrus.

Animals↗

Different doses of porcine luteinizing hormone in precocious puberty induction in gilts.

The use of hormonal protocols in puberty induction and synchronization of oestrus has lead to an increase in the efficiency of replacement gilts. The aim of this study was to evaluate different doses of porcine LH in precocious puberty induction and oestrus synchronization in a homogeneous group of gilts. Sixty-seven homogeneous prepubertal gilts (Camborough 22) at 137 +/- 4 days of age and 87 +/- 7 kg were treated with three different hormonal protocols: T1--600 UI of equine chorionic gonadotrophin (eCG; Novormon) and after a 72-h period 5 mg of LH (Lutropin); T2--600 UI of eCG and a 72-h period 2.5 mg of LH; T3--600 UI of eCG and a 72-h period 1.25 mg of LH. The ovaries were examined at slaughter, on day 6 after the hormonal treatment. There were no statistical differences (p > 0.05) between the different LH doses in the percentage of the detected oestrus (T1 = 42.85%; T2 = 60.87%, T3 = 52.18%), oestrus duration (T1 = 41.44 +/- 16.30 h; T2 = 48.57 +/- 16.29 h, T3 = 39.33 +/- 11.42 h), number of corpora lutea (T1 = 9.61 +/- 5.43; T2 = 9.86 +/- 3.32, T3 = 8.13 +/- 5.52) and percentage of animals presenting ovarian cystic degeneration (T1 = 33.33%; T2 = 39.13%, T3 = 39.13%). The T2 (2.5 mg of LH) presented the lowest dispersion (p < 0.05) of the LH-ovulation interval (T1 = 37.17 +/- 4.07 h; T2 = 38.26 +/- 2.84 h; T3 = 36.25 +/- 5.69 h). The LH dose reduction to 2.50 and 1.25 mg presented equal results with the recommended dose of 5.0 mg, and could be used in the precocious induction of oestrus in gilts. The 2.5-mg LH dose showed the lowest dispersion of ovulation and it can be used in fixed-time artificial insemination programmes.

Animals↗

Proteolysis of insulin-like growth factor binding proteins during preovulatory follicular development in cattle.

The present study was conducted to evaluate changes in follicular fluid (FF) insulin-like growth factor binding protein (IGFBP) proteolytic activity and levels of steroids and IGFBP during follicular development in cattle. Estrous cycles of cows were synchronized with two injections of prostaglandin F2alpha (PGF) 11 d apart and follicular growth monitored via daily rectal ultrasonography in order to identify the dominant follicle. All cows were ovariectomized 48 hr after the second injection of PGF. Follicular fluid was collected individually for all follicles > 5 mm and pooled for small (1 to 5 mm) follicles. Follicular fluid estradiol and androstenedione levels were greater (P < 0.05) and progesterone and IGFBP-3 levels not different (P > 0.10) in large dominant than in small (1 to 5 mm) or large (>5 mm) subordinate follicles, whereas IGFBP-2, -4 and -5 levels were less (P < 0.05) in large dominant than in small or large subordinate follicles. To evaluate proteolysis of IGFBPs, FF was incubated with recombinant human (125) I-labeled IGFBP-2, -3, -4, and -5 and proteins separated by 12% SDS-PAGE. Follicular fluid caused little or no proteolysis of (125)I-lableled IGFBP-2 or -3. However, cleavage of (125)I-labeled IGFBP-4 and -5 by FF from large dominant follicles was greater (P < 0.05) than by FF from small or large subordinate follicles indicating that a protease to IGFBP-4 and -5 exists in estrogen dominant follicles. We conclude that lower levels of IGFBP-2 in estrogen dominant follicles of cattle are not due to increased proteolysis, whereas decreases in IGFBP-4 and -5 levels are likely due, in part, to increased protease activity. Changes in IGFBP may alter levels of bioavailable IGFs that stimulate steroidogenesis and mitogenesis in developing bovine follicles.

Androstenedione↗

Excess dietary urea intake in ewes and its effect on ovulation rate and embryo development.

The effect of excess dietary urea on ovulation and early embryo development of sheep was studied. Thirty Border Leicester x Scottish Black face ewes randomly assigned to three treatments were given a basal control diet (C) which met energy requirements for body weight maintenance. Other treatments were basal diet plus 24 g of urea/day (low urea, L) or plus 48 g (high urea, H)/day. The reproductive cycles of the ewes were synchronized using a single injection of prostaglandin (PGF2 alpha) and progesterone by an intravaginal controlled internal drug release (CIDR) device for 12 days. Ovulation was induced by the use of pregnant mare serum gonadotrophin (PMSG). Ewes were inseminated approximately 52 hours after CIDR device removal using a laparoscopic technique. Embryos were recovered at Day 4 or Day 11 after insemination from half of the ewes from each treatment group. There were no significant differences in ovulation rates among the three groups. The embryo recovery rates were not affected by day of recovery. At embryo recovery on Day 4, 7/13 in C, 3/6 in L and 0/7 in H embryos were morulae. After 72 hours of in vitro culture 6/10 in C, 2/3 in L and 0/4 in H embryos developed to the blastocyst stage. Pregnancies sustained were C 6/8, L 5/7 and H 1/3 of the autotransfers. Throughout the experiment plasma urea levels were significantly affected by diet (p < 0.01). Plasma ammonia levels in the H group were significantly higher than those in the C and L groups (p < 0.05) for 4 hours after each feed. There was no treatment effect on plasma progesterone concentration. The luteinizing hormone (LH) surge onset time and amplitude were not correlated to ovulation rate and were not affected by treatment. It is concluded that high circulating concentrations of plasma urea and ammonia have an adverse effect on early embryo development. This effect was independent of any alterations in progesterone and LH concentrations.

Ammonia↗

Preovulatory follicle development and ovulation in the brushtail possum (Trichosurus vulpecula) monitored by repeated laparoscopy.

The common method for synchronizing oestrus in brushtail possums is by removal of their pouch young (RPY). However, there is little information on the ovarian response to this treatment, the timing and incidence of ovulation is poorly defined, and methods of identifying oestrus are unreliable. In this study, the development of preovulatory follicles, ovulation and reproductive tract changes following RPY were monitored by repeated laparoscopic observation. A total of 120 adult female possums underwent laparoscopy at intervals of 1-4 days over the period from 0 to 21 days after RPY. Tissue was collected from a further 30 animals for correlative histology of ovarian structures, and to quantify changes in reproductive tract organs. Only 80 of 120 animals ovulated, and the time of ovulation ranged from 7 to 18 days following RPY. In most animals, enlargement of vaginal cul-de-sac and uterine tissue occurred within 10 days. Correlative histology supported the macroscopic classification of ovarian structures, and healthy and atretic follicles could be identified by laparoscopy. Vaginal smears and plasma progesterone concentrations verified the occurrence of ovulation as observed by laparoscopy. A 'presumptive' preovulatory follicle, first identifiable approximately 5 days before ovulation, was recorded in all animals that ovulated and in none that failed to ovulate. Changes to its surface morphology indicated impending ovulation. This study has enabled the day of ovulation to be identified accurately for the first time in this species. It has also shown that there is wide variation in follicle development, lack of synchrony in the time of ovulation in the brushtail possum, and that some animals fail to ovulate following RPY.

Animals↗

Inhibition of intrafollicular PGE2 synthesis and ovulation following ultrasound-mediated intrafollicular injection of the selective cyclooxygenase-2 inhibitor NS-398 in cattle.

Ultrasound-mediated intrafollicular injection and aspiration procedures were used to investigate the ability of the selective cyclooxygenase-2 inhibitor, NS-398, to inhibit intrafollicular PGE2 synthesis and suppress ovulation in dairy cattle. Follicular growth and timing of the preovulatory gonadotropin surge were synchronized in 55 Holstein cows and the position of the ovulatory follicle was determined by daily ultrasound scanning. Preovulatory follicular fluid was aspirated from the largest follicle in four animals at 0, 6, 12, 18, and 24 h after GnRH injection (n = 20). The remaining 35 animals were subjected to ultrasound-mediated intrafollicular injection of NS-398 (10 microM final concentration; n = 19) or diluent (n = 16; controls). At 24 h after GnRH injection, follicular fluid was harvested from a subset of NS-398- (n = 9) and diluent-treated animals (n = 6). The remaining NS-398- and diluent-treated animals were subjected to ultrasonography every 6 h for 36 h after intrafollicular injection, and then daily through d 7 of the subsequent luteal phase to monitor ovulation and corpus luteum development. Follicular fluid PGE2 concentrations were increased following GnRH injection and reached a maximum at 24 h (P < 0.05). Follicular fluid PGE2 concentrations were decreased in NS-398- vs. diluent-treated follicles (7.2 vs. 52.2 ng/mL respectively; P < 0.05), but progesterone concentrations did not differ. Intrafollicular injection of NS-398 also inhibited follicle rupture (P < 0.001). All 10 control animals ovulated within 30 h of GnRH injection. Nine out of the ten NS-398-injected animals failed to ovulate. The NS-398-injected follicles developed morphological and endocrine characteristics resembling luteinized, unruptured follicles. Thus, intrafollicular PGE2 synthesis and follicle rupture, but not luteinization, were inhibited in cattle following ultrasound-mediated intrafollicular injection of NS-398. Ultrasound-mediated intrafollicular injection of NS-398 is a useful tool for mechanistic studies of intrafollicular regulation of the ovulatory process in cattle.

Animals↗

Changes in the level of endogenous leptin, FSH, 17beta-oestradiol and metabolites during lupin-induced increase in ovulation rate in ewes.

The aim of the study was to determine the changes in the plasma concentration of leptin during lupin feeding-induced increase in the ovulation rate (OR) in ewes. Additionally, alterations in the plasma level of glycogenic amino acids and glucose (as the factors influencing leptin secretion) and the levels of follicle-stimulating hormone (FSH) and 17beta-oestradiol (E-2) (as the hormones regulated by leptin and engaged in recruitment, selection and development of ovulatory follicles) were analysed. Ninety-six female Polish Lowland Sheep were used. All ewes were cyclic and synchronized with PGF2alpha. The ewes were divided into two groups: control (n = 48), fed only with hay, and experimental (n = 48), received additionally lupin (Lupinus angustifolius) grain as a high-protein and a high-energy supplement. They were given lupin from the second to 13th day of the oestrous cycle at increasing doses (150-750 g/day per ewe). On the 11th day of cycle blood samples for analysis of hormones, amino acids and total glucose concentration, were collected from the jugular vein. OR was determined by laparoscopy of ovaries on the sixth day of the following oestrous cycle. Mean OR of ewes supplemented with lupin grain (1.687 +/- 0.463) was 30.67% higher than that of control (1.291 +/- 0.454). In spite of the unchanged body mass, a significant increase (P < or = 0.05) in mean concentration of plasma leptin in the experimental ewes [2.17 +/- 0.15 ng/ml human equivalent (HE)] was found in comparison with control (1.42 +/- 0.12 ng/ml HE). A significantly (P < or = 0.05) higher plasma FSH level in the ewes fed lupin (105.21 +/- 5.87 ng/ml) compared with those fed hay (67.88 +/- 6.03 ng/ml) was also found. However, plasma level of E-2 decreased after lupin feeding. Moreover, in the ewes fed lupin the plasma concentrations of glucose and nine glycogenic amino acids (Gly, Ala, Val, Met, Leu, Ile, Tyr, Phe and Arg) were increased. It can be concluded that lupin feeding exerts the stimulatory effect on the OR in Polish Lowland Sheep. The increase in OR is connected with significantly higher plasma leptin level and coincident with rise in FSH, glycogenic amino acids and glucose concentration. In contrast, the level of plasma E-2 was significantly decreased in lupin-fed ewes.

Amino Acids↗

Successful intrauterine insemination of Eld's deer (Cervus eldi thamin) with frozen-thawed spermatozoa.

This study tested the efficacy of assisted reproduction (synchronization of oestrus and intrauterine artificial insemination (AI)) in contributing to the captive propagation of an endangered species, the Eld's deer (Cervus eldi thamin). Semen was collected from males preselected on the basis of under-represented genotype. Motility of spermatozoa after thawing from ejaculates diluted with BF5F extender (8% glycerol), frozen on dry ice in 0.5 ml straws and stored in liquid nitrogen was 60-70%. Intravaginal progesterone-releasing devices (controlled internal drug release, CIDR-type G) were inserted into 20 adult Eld's deer hinds for 14 days. In all hinds, semen (7.5-10 x 10(6) motile spermatozoa per uterine horn) was deposited by laparoscopy performed 70 h after removal of the CIDR device. Ovarian activity, before and after AI, was monitored by analysing pregnanediol-3 alpha-glucuronide (PdG) concentrations in voided urine collected three to seven times per week. During the period of CIDR device insertion, urinary PdG profiles were equal to, or above, normal luteal phase concentrations in all hinds. Within 48 h of device withdrawal, PdG concentrations returned to baseline values in 17 of the 20 females, and the onset of behavioural oestrus occurred at this time in 12 hinds. On the basis of sustained increases in urinary PdG, 9 of the 20 hinds were diagnosed as pregnant by 90 days after AI, all of which delivered offspring after a mean gestation of 241.1 days (range, 235-245). Seven singletons (two females, five males) were born alive and survived, and one singleton and one set of twins were stillborn (three females).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The effect of increased dietary intake on superovulatory response to FSH in heifers.

We have previously shown that the number of ovarian follicles <4 mm in diameter can be increased by enhanced dietary intake in heifers. This study investigated the effect of the same dietary treatment on superovulatory response. The estrous cycles of 24 mature Hereford x Friesian heifers were synchronized by a standard progesterone plus prostaglandin protocol. The animals were fed with either 100% (group M, n = 12) or 200% (group 2M, n = 12) maintenance requirements for a 3-week period. Starting from day 4 of the synchronized estrous cycle, all the animals were superovulated using a standard 4-day FSH regime followed by an injection of GnRH analogue (GnRHa) to induce ovulation. Rectal ultrasound scanning was carried out to assess ovarian follicular populations at the start of FSH treatment and on the day of GnRHa injection, and to determine the number of corpora lutea 5 days after GnRHa injection. The body weight (BW) and body condition score (BCS) were recorded weekly and plasma samples were collected throughout the experimental period. There were no differences in either BW or BCS between two groups at the start of the experiment. The BW and BCS were maintained during the experiment in the group M, whilst animals in the group 2M showed a non-significant (P > 0.05) increase in BW and BCS. Circulating concentrations of insulin were significantly (P < 0.01) higher in heifers from the group 2M throughout the controlled feeding period. The group 2M had significantly (P < 0.05) more follicles 2-4 mm in diameter at the start of FSH treatment and more (P < 0.01) follicles >9 mm in diameter on the day of GnRHa injection, when compared with the group M. Similarly, 5 days after GnRHa injection there were significantly (P < 0.01) more corpora lutea in the group 2M (18.1+/-2.2) than in the group M (10.6+/-3.0). In addition, plasma progesterone concentrations following GnRHa injection were significantly (P < 0.01) higher in heifers from the group 2M. In conclusion, these results confirm that increased dietary intake can enhance the recruitment of ovarian follicles in heifers. This treatment may provide a valuable approach to improving superovulatory response in cattle.

Animals↗

Serum progesterone levels in post-partum dairy cows after repeated application of the prostaglandin F2 alpha analogue D (+) cloprostenol sodium.

Reproductive management programmes based on strategic use of prostaglandin F2 alpha (PGF2 alpha) to induce and synchronize oestrus in post-partum dairy cows are widespread. Repeated shortening of the oestrous cycle during early lactation in high-yielding dairy cows, however, could impair corpus luteum function and thus decrease fertility. The objective of this study was to analyse the effect of repeated treatments with the prostaglandin F2 alpha analogue D (+) cloprostenol sodium on progesterone concentrations indicative of a functional corpus luteum in post-partum dairy cows. Furthermore, the influence of milk production, parity and endometritis on progesterone concentrations under these circumstances were studied. Eighty-four cows of a commercial dairy operation were treated three to four times with D (+) cloprostenol sodium (Preloban; Hoechst Roussel Vet, Wiesbaden, Germany) at 14-day intervals, starting 22-28 days post-partum. Blood samples were collected prior to treatment 1 (sample 1) and 14 days after treatments 1, 2 and 3 (samples 2-4) and serum progesterone (P4) levels were determined. The percentage of cows with P4 levels < 1 ng/ml decreased from 51% in sample 1 to 23% in samples 3 and 4. More primiparous cows had low P4 levels 14 days after the second treatment than older cows (P < 0.05). Cows with low progesterone levels in sample 3 or 4 had lower protein contents in milk on the second milk test day post-partum and in their cumulative milk yield of the first 100 days of lactation. Clinical endometritis at post-partum examination did not influence progesterone levels after treatment with PGF2 alpha. Repeated application of PGF2 alpha (more than twice) in the post-partum period does not influence serum progesterone levels 14 days after treatment. Failure to develop luteal tissue after treatment contributed to the conception failures after first service.

Animals↗

A coupled-oscillator model of ovarian-cycle synchrony among female rats.

The ovarian cycles of female rats become synchronized when they live together, as do the cycles of many other mammals. Ovarian cycles also become synchronized when rats live apart if they share a common air supply, indicating that ovarian-cycle synchrony is mediated by pheromones. We developed a coupled-oscillator model of ovarian-cycle synchrony to test several hypotheses about its pheromonal and neuroendocrine mechanisms and to guide our experimental research. The model spans three levels of organization: the group, the rat, and the neuroendocrine components of the ovarian system. The ovarian system (not the ovaries themselves) are modeled as an oscillating system. Coupling among ovarian systems is mediated by the exchange of two pheromones, one that delays the phase of the ovarian system and one that advances it. Computer simulation experiments showed that this coupled-oscillator model can explain the levels of ovarian-cycle synchrony observed in groups of female rats while, at the same time, matching an empirical distribution of ovarian-cycle lengths. By successfully matching computer simulation data with empirical data, we were able to infer theoretical predictions in a number of areas: (1) effect of initial conditions on the probability that a group will change to different synchrony level and phase relationships, i.e. the transition probability between all synchrony levels and phase relationships; (2) effects of individual differences in pheromone sensitivity on ovarian-cycle synchrony; (3) the timing of pheromone sensitivity during the ovarian cycle; and (4) the existence of partial luteinizing hormone surges, which may cause the "spontaneous" prolonged ovarian cycles associated with ovarian-cycle synchrony. The paper concludes by discussing the integrative role of this model for experimental research. In particular, we focus on the role of this model in interpreting theoretical aspects of ovarian-cycle synchrony as well as for guiding future experimental research into its mechanisms and functions.

Animals↗

Effect of season and gonadotropins on the superovulatory response in camel (Camelus dromedarius).

The purpose of the present investigation was to study the extent to which season and the gonadotropin preparation interferes with the superovulatory response in the dromedary. Adult camels were treated for superovulation during the breeding (November to April) and non-breeding season (May to October). Animals were synchronized by daily i.m. injections of progesterone (125 mg/animal/day, Jurox, UK) for 10 to 14 days. Superovulation was induced by 400mg pFSH alone (Follitropin V, Vetrepharm, Canada) administered in eight descending doses at 12h intervals or a combination of PMSG (2000IU, Folligon, Intervet, The Netherlands), injected with last injection of progesterone and 400mg pFSH in eight descending doses. The follicular development was daily assessed by ultrasonography of the ovaries. The donors were classified as per their response to the superovulatory treatment into very good (>10 follicles), good (5-10 follicle), poor (2-4 follicles) or no response (1 or no follicle) on each ovary. Ovulation was induced by injecting 3000 IU hCG (Chorulon, Intervet) at the time of first mating. The donors were mated twice at an interval of 12h when all or most of the follicles reached to a size of about 1.0-1.7 cm. Camels were flushed non-surgically on Day 6 or 7 after the ovulation. The proportion of camels showing very good response during the breeding as well as non-breeding season was higher (P<0.05) when a combination of pFSH and eCG was used compared with pFSH only. There was no difference (P>0.05) in the proportion of donors flushed successfully (embryos recovered) when treated either with a combination of pFSH and eCG or pFSH alone during the breeding and non-breeding season. The rate of recovery of ova/embryos and proportion of transferable embryos was higher (P<0.05) when donors were treated with pFSH+eCG compared with pFSH only during the breeding as well as non-breeding season. The results may indicate that ova/embryo recovery rate of the dromedary is influenced by the gonadotropin preparation but is not appreciably affected by the season.

Animals↗

Effects of bromocriptine administration during the follicular phase of the oestrous cycle on prolactin and gonadotrophin secretion and follicular dynamics in merino monovular ewes.

Two experiments using Spanish Merino ewes were conducted to investigate whether the secretion of prolactin during the follicular phase of the sheep oestrous cycle was involved in the patterns of growth and regression of follicle populations. In both experiments, oestrus was synchronized with two cloprostenol injections which were administered 10 days apart. Concurrent with the second injection (time 0), ewes (n = 6 per group) received one of the following treatments every 12 h from time 0 to 72 h: group 1: vehicle injection (control); group 2: 0.6 mg bromocriptine (0.03 mg per kg per day); and group 3: 1.2 mg bromocriptine (0.06 mg per kg per day). In Expt 1, blood samples were collected every 3 h from 0 to 72 h, and also every 20 min from 38 to 54 h to measure prolactin, LH and FSH concentrations. In Expt 2, transrectal ultrasonography was carried out every 12 h from time 0 until oestrus, and blood samples were collected every 4 h to measure prolactin, LH and FSH concentrations. Ovulation rates were determined by laparoscopy on day 4 after oestrus. Bromocriptine markedly decreased prolactin secretion, but did not affect FSH concentrations, the mean time of the LH preovulatory surge or LH concentrations in the preovulatory surge. Both doses of bromocriptine caused a similar decrease in LH pulse frequency before the preovulatory surge. The highest bromocriptine dose led to a reduction (P < 0.01) in the number of 2-3 mm follicles detected in the ovaries at each time point. However, bromocriptine did not modify the total number or the number of newly detected 4-5 mm follicles at each time point, the number of follicles > 5 mm or the ovulation rate. In conclusion, the effects of bromocriptine on gonadotrophin and prolactin secretion and on the follicular dynamics during the follicular phase of the sheep oestrous cycle indicate that prolactin may influence the viability of gonadotrophin-responsive follicles shortly after luteolysis.

Analysis of Variance↗

Time and incidence of ovulation and conception rates after incorporating estradiol cypionate into a timed artificial insemination protocol.

Two experiments were conducted to determine the effect of estradiol cypionate (ECP), when incorporated into a conventional GnRH-PGF(2alpha)-GnRH timed artificial insemination protocol (Ovsynch), on systemic estradiol (E(2)), time and incidence of ovulation, luteal development, and conception rate in Holstein cows. Our objective was to determine if administration of 0.25 mg of ECP at the time of the second GnRH injection would effectively synchronize ovulation and increase conception rate. In Experiment 1, lactating Holstein cows (n = 23; 58.7 +/- 1.2 d in milk) were synchronized with PGF(2alpha) (at d -10). Ten days later, Ovsynch was initiated with the administration of 100 mug of GnRH (d 0) followed by PGF(2alpha) on d 7. On d 9, cows were assigned randomly to be treated with either GnRH + 0.25 mg of ECP (OVS-ECP; n = 11) or GnRH and 1 mL of cottonseed oil (OVS-C; n = 12). Ovarian activity was monitored by ultrasonography on d 0, 7, and 9. To determine the time of ovulation, ultrasound examinations were conducted at 12 and 20 h posttreatment and then at least every 3 h until either 36 h posttreatment or ovulation was observed. Blood samples were collected on d 0, 7, 9, and 16 for progesterone analysis. Blood samples also were collected at the time of treatment (d 9, 0 h) and at 6, 12, 20, and 28 h for E(2) analysis. Incidence of ovulation did not differ between treatments. Mean ovulation time relative to the second GnRH administration was similar between treatments. Serum progesterone concentration did not differ between treatments at any time. Serum E(2) concentration was not different at the time of treatment (0 h); however, mean E(2) concentration was greater for the OVS-ECP group at 6 and 12 h after treatment compared with OVS-C. In Experiment 2, lactating dairy cows (n = 333) in 3 commercial herds were randomly assigned to OVS-ECP (n = 169) or OVS-C (n = 164). Cows were inseminated 22 to 24 h posttreatment. Conception rates did not differ between treatments. Estradiol cypionate treatment was successful in increasing serum E(2) when administered at the time of the second dose of GnRH in the Ovsynch protocol. Conception rates, however, were not affected by treatment.

Animals↗

Effect of level of dietary energy and protein on embryo survival and progesterone production on day eight of pregnancy in Rasa Aragonesa ewes.

The aim of this experiment was to investigate the effects of dietary protein and energy on ovulation rate and embryo survival to day 8 of pregnancy, and the associated concentrations of progesterone in jugular, ovarian and uterine veins, in a Spanish breed of sheep. In mid-October, three groups of ewes were fed to provide 1.5 x (H; n = 9), 0.5 x (L; n = 12) or 0.5 x plus 7.44 g CP/MJ ME (LP; n = 8) energy requirements for maintenance of live weight from day -14 relative to a synchronized mating on day 0. A significant effect of nutrition on ovulation rate was observed (H: 2.22 +/- 0.16; L: 1.50 +/- 0.16; LP: 1.88 +/- 0.12 corpora lutea; P < 0.05). Mean LH and progesterone concentrations were affected by nutrition on day 7, L ewes showing the highest mean LH level (P < 0.01), while H ewes presented the lowest mean LH concentration and the highest mean plasma progesterone concentration (P < 0.01). Laparotomies were performed on six animals of each group on day 8 to determine the effect of nutrition on embryo development. A significantly higher percentage of embryos recovered from L and LP ewes presented an earlier stage of development (morulae or early blastocysts) (P < 0.001), while 100% embryos of H ewes were expanded blastocysts. The ratio expanded blastocysts/corpora lutea was significantly higher in H ewes (0.86) when compared with L and LP groups together (0.57; P < 0.05). Mean progesterone concentration in the ovarian vein was 800-fold higher than mean jugular venous levels with no differences between groups. Samples from ovarian veins contralateral to corpus luteum-bearing ovaries showed mean progesterone concentrations significantly lower than samples opposite to corpus luteum (ipsilateral: 1037.84 +/- 138.45; contralateral: 30.4 +/- 11.22 ng/ml; P < 0.001). Mean progesterone concentration in the uterine vein was approximately 30-fold higher than in jugular and similar in both uterine horns and treatments. No effect of nutrition on pregnancy rate was observed (H: 89%; L: 92%; LP: 100%). These results suggest that neither dietary energy nor protein are able to modify pregnancy rate or progesterone concentrations in ovarian and uterine veins eight days after mating. However, the delay in embryo development observed in the embryos collected from L and LP ewes may give rise to compromised embryo growth and development some days later.

Animal Nutritional Physiological Phenomena↗

Differences between Belclare and Suffolk ewes in fertilization rate, embryo quality and accessory sperm number after cervical or laparoscopic artificial insemination.

Ewe breed has been shown to have a major effect on pregnancy rates following cervical AI using frozen-thawed semen. The main objective of this study was to examine the differences between purebred Belclare and Suffolk ewes (multiparous) in fertilization rate, number of accessory sperm and stage of embryo development on day 6 after cervical or laparoscopic AI with frozen-thawed semen. In experiment 1, Belclare and Suffolk ewes were synchronized for 12 days and were either cervically inseminated (year 1: n=28 and 31; year 2: n=16 and 15, respectively) or laparoscopically inseminated (year 2: n=13 and 14). In experiment 2, superovulated Belclare (n=4) and Suffolk (n=13) ewes were laparoscopically inseminated. All ewes were slaughtered 6 days after AI; oocytes/embryos were recovered, morphologically graded and stained to assess the number of cells and accessory spermatozoa. Data from both experiments were combined for statistical analysis. The proportion of ewes with fertilized oocytes was significantly higher following laparoscopic AI compared with cervical AI (54% versus 19%). More Belclare than Suffolk ewes yielded fertilized oocyte(s) after cervical AI (34% versus 10%, P<0.02) but there was no difference after laparoscopic AI (62% versus 60%). From the ewes that yielded at least one fertilized oocyte the proportion of Belclare ewes with embryos at the morula/blastocyst stage was significantly greater than for Suffolk ewes (94% versus 59%, P<0.02). A higher proportion of Belclare than Suffolk ewes had evidence of sperm reaching the site of fertilization following cervical AI (39% versus 15%, P<0.02) but there was no difference after laparoscopic AI (62% versus 64%, P>0.8). Amongst the ewes with evidence of sperm at the site of fertilization, laparoscopic AI resulted in a higher number of sperm per oocyte/embryo or per ewe than cervical AI (P<0.01). These results suggested that the difference in pregnancy rate between Suffolk and Belclare ewes following cervical AI was due to: (i) sperm traversing the cervix and uterus in a higher proportion of Belclare than Suffolk ewes, leading to a higher incidence of fertilization and (ii) the lower developmental competence of fertilized oocytes from Suffolk ewes.

Animals↗

Dexamethasone influences endocrine and ovarian function in dairy cattle.

Multiparous nonlactating Holstein cows were used to determine the effect of dexamethasone on ovarian follicular development and plasma hormone concentrations. Animals were randomly divided into two groups, control (C; n = 5) and treatment (T; n = 6), but managed as one group. Both groups were synchronized with two injections of PGF2alpha (25 mg i.m.) 11 d apart. One day after ovulation (d 0) the T group received a daily injection of dexamethasone (44 microg/kg of body weight; i.m.) until the first dominant follicle stopped growing or up to d 12 postovulation. The C group received vehicle injections. Blood samples were collected daily from all cows. Concentrations of LH and FSH did not differ between the C and T cows, whereas progesterone concentrations were lower in T than in C cows from d 4 onward. Treatment x day interaction influenced plasma insulin concentrations such that T cows had insulin concentrations 2.9- to 6.0-fold those of C cows between d 2 and 9. Dexamethasone decreased IGF-I and -II concentrations from d 5 onward. Concentrations of plasma leptin and the various IGF binding proteins were not affected by dexamethasone. Total number of follicles (> or = 5 mm) and plasma estradiol concentrations were less in T than in C cows on d 0, 1, and 4. The growth rate of the dominant follicles and maximum diameter of the dominant and subordinate follicles were not affected by dexamethasone. The diameter of the CL was 21 to 39% larger in T than in C cows between d 6 and 10. Treatment x day interaction influenced plasma cholesterol concentrations such that cholesterol levels decreased 46.8% in T cows and 19.5% in C cows between d 0 and 10. Plasma glucose concentrations were greater in T than in C cows between d 1 and 10. In summary, dexamethasone had significant effects on metabolism without a major impact on growth of the first-wave dominant follicle. Dexamethasone-induced suppression of luteal function was associated with decreased plasma IGF-I and -II concentrations.

Animals↗

[Synchronization of ovulation (OVSYNCH) in high producing dairy cattle herds. Part II: Milk yield, diseases and cost-benefit-analysis].

A method for synchronization of ovulation (OVSYNCH) was evaluated for its practical use in seven North German dairy cattle herds. The original procedure of PURSLEY et al. (1995) was applied, i. e. cows were treated with Gonadotropin Releasing Hormone (GnRH) at a random stage of the estrous cycle, followed by an injection of a prostaglandin F2alpha (PGF2alpha)-analog seven days later and another injection of GnRH two days after PGF2alpha. Cows were inseminated 24 h after the last injection by artificial insemination. Control animals were assigned by corresponding lactation number and month of lactation. Cows from the OVSYNCH group returned to the conventional reproductive management after the OVSYNCH treatment, which was the same management as that for the control group. The parameters of fertility and correlations between body condition score (BCS) as well as concentrations of serum progesterone and the success of OVSYNCH were presented in the first report. For the OVSYNCH group, the 100-day-duction during the time period from calving until a pregnancy was established differed markedly from healthy cows in a variety of parameters. Healthy cows had a greater FSC (65.22%) than sick cows (35.90%) (P < 0.05). Furthermore, mean days open were less in healthy cows than in sick cows (85.48 +/- 30.56 days vs 110.85 +/- 46.40 days) (P < 0.05). Cows showing a disease had a higher number of lactation (4.00 +/- 2.26 vs. 2.80 +/- 1.35; P < 0.01) and a decreased BCS (2.72 +/- 0.23 vs. 2.91 +/- 0.23; P < 0.05). Sick and healthy cows from the OVSYNCH group did not differ with regard to 100-day-milk-yield (milk kg, relative and absolute milk fat and protein content). The cost-benefit-analysis revealed higher total cost, and cost per pregnancy (16,408.07 euros and 278.10 euros, respectively) in the control group than in the OVSYNCH group (11,015.69 euros and 196.71 euros, respectively).

Animals↗