Effect of site of deposition on the fertility of sheep inseminated with frozen-thawed semen

The widespread use of artificial insemination (AI) in sheep is currently prevented due to the lack of a cost effective insemination technique utilising frozen-thawed semen. The objective of the present study was to determine if the deposition of frozen-thawed semen in the vaginal fornix would result in a pregnancy rate comparable to that achieved following cervical insemination. Multiparous ewes of various breeds were synchronised and inseminated into either the vaginal fornix (n=78) or the cervix (n=79), at 57 h post sponge removal, with frozen-thawed semen. Information on mucus secretion and the depth to which it was possible to penetrate the cervix at insemination (cervically inseminated ewes only) was recorded at the time of AI. Pregnancy rate was subsequently determined either by return to service (oestrus) or after slaughter 30 days post insemination. Insemination site did not significantly influence pregnancy rate using frozen-thawed semen (36.2% compared to 27.6% for cervical and vaginal fornix insemination, respectively; P=0.26). Whilst depth of cervical penetration was positively associated with pregnancy rate (P<0.05), this association needs to be interpreted with caution as none of the ewes where the cervix could not be penetrated (score=0) was pregnant. In conclusion, pregnancy rate following insemination of frozen-thawed semen into the vaginal fornix was within 10% points of that obtained following cervical AI of frozen-thawed semen. As insemination into the vaginal fornix is technically easier than cervical insemination, it may be more practical for use in large scale applications.     

Lack of an effect of prostaglandin injection at estrus onset on the time of ovulation and on reproductive performance in weaned sows

We evaluated the effects of a PGF2alpha analogue on time of ovulation and reproductive performance in multiparous Camborough sows (n=47). At onset of first post-weaning estrus, sows received either an intravulval injection of 3.75 mg of prostaglandin analogue (PGF) or, served as a non-injected control (CON). Beginning 24 h after the onset of estrus, transcutaneous ultrasonography was carried out every six h to determine time of ovulation. At 36, 54, and 72 h after the onset of estrus, blood samples were taken for progesterone analysis. Weaning-to-estrus (WEI), duration of estrus, ovulation rate and number of live embryos at d 28 of gestation were recorded. Treatment had no effect (P > 0.05) on any parameters measured. Duration of estrus classified as less or greater than the overall mean also had no effect (P > 0.05) on any of the parameters measured. Results indicate that treatment did not advance ovulation nor did it improve reproductive performance in sows. Overall, a negative correlation of WEI with the ovulation rate (P = 0.0005, r = -0.54) was established.     

Effect of sterile service on estrus duration, fertility and prolificacy in artificially inseminated dairy goats

The effect of sterile service on estrus duration, fertility and prolificacy in artificially inseminated dairy goats during breeding season was studied. Nubian does (n=126) were divided into 2 equal groups: service and control. Estrus was synchronized with intravaginal sponges containing either fluorgestone acetate (FGA; 40 mg) or medroxiprogesterone acetate (MAP; 60 mg) for 12 or 14 d, respectively. Two vasectomized teaser bucks were used to detect estrus at 6-h intervals for 5 d after sponge removal (0600, 1200, 1800 and 2400 h). The teasers were fitted with aprons and permitted to mount all does in both groups, but to penetrate only the service does within the first 12 h of estrus. Does in both groups were inseminated twice at 12 and 24 h after estrus was first detected, using 1 straw per insemination containing 200 million of cooled spermatozoa from 1 buck. The semen was placed in mid-cervix. Estrus duration for the service and control does was (mean +/- SD) 29.4 +/- 6.5 and 41.8 +/- 9.6 h, respectively. Fertility for the service does was 73.7% (46/63); for control does it was 58.7% (37/63). Prolificacy was 2.1 (96/46) and 2.0 (74/37) for service and control does, respectively. Estrus duration (P<0.001) and fertility (P<0.05) differed between the service and control group, but prolificacy was similar (P>0.05). It is concluded that sterile service reduces the duration of estrus and increases fertility in artificially inseminated dairy goats.     

Some effects of the composition of inseminated semen and the site of its deposition on fertility in Gallus domesticus

It is most probable that during natural copulation the semen of the fowl is ejaculated into a shallow position in the vagina of the hen, but during the commercial application of artificial insemination it is generally considered necessary to evert the distal vagina and deposit semen to a depth of at least 5 cm to produce optimal fertilisation of the succession of eggs laid daily by a female for a week post-insemination. Aspects of the artificial insemination technique in relation to the types of semen that are obtained from the male fowl artificially are re-appraised in relation to their effect on fertility. It was confirmed that a smaller number of spermatozoa (50 × 10⁶) than is normally used in commercial practice (>80 × 10⁶) produced good fertility, even when inseminated within 0.5 cm of the vaginal opening in the cloaca. The results were achieved whether or not glucose was present in the inseminate. When semen was deposited in the cloaca, a better fertilisation rate was obtained if ductus-deferens semen was diluted with transparent fluid, which is produced by tumescent tissue in the cloaca during semen collection. However, the same advantageous effect was shown by dilution with synthetic aqueous fluids with and without glucose. The likely role of transparent fluid during natural copulation is discussed. On the basis of the number of spermatozoa found to maintain good fertility by artificial insemination, only 10 μl semen would be required to be ejaculated into each hen during copulation. This may account for the well-known ability of the male fowl to copulate frequently in a day, because the small volume of semen would be replenished, naturally, very quickly in the ductus deferens.     

Effect of PG600 and adjusted mating times on reproductive performance in weaned sows

The administration of PG600 to sows at weaning induces >90% of sows to return to estrus within a week, but farrowing rate and litter size are often not improved. This study evaluated the effects of adjusted artificial insemination (AI) times based on weaning to estrus interval (WEI) and estrus to ovulation interval (EOI) following PG600. All sows were given PG600 at weaning and allotted to adjusted (ADJ, n=47) or non-adjusted (NA, n=46) mating times after the onset of estrus. Adjusted mating involved: (1) 2-3 days WEI, AI at 36 h and 48 h; (2) 4 days WEI, AI at 24h and 36 h; (3) 5 days WEI, AI at 12h and 24h; and (4) 6-7 days WEI, AI at 0 h and 12h. Mating for NA occurred at 0 h and 24h after onset of estrus. There was no effect of treatment on return to estrus (92.9% versus 92.5%) or ovulation (92.7% versus 92.5% for ADJ and NA, respectively). The proportion of first AI occurring within 24h prior to ovulation was increased (83.8% versus 50.0%) and closer to ovulation for ADJ compared to NA treatment (19.4h versus 27.3h, P<0.05). Treatment did not influence (P>0.10) the proportion of second AI occurring within 24h of ovulation (72.8% versus 56.6%) but did influence (P<0.05) the interval from second AI to ovulation for ADJ compared to NA (10.6h versus 3.3h). The ADJ treatment increased (P<0.05) the proportion of sows that received an AI within 24h before ovulation (98.8% versus 87.0%). However, treatment did not influence pregnancy (87.4%) or farrowing (79.5%) rates but the NA treatment tended to increase (P<0.10) total number of pigs born (11.8 versus 8.9). In conclusion, while AI times for ADJ appeared to occur within optimal periods, farrowing rates were not improved and litter size decreased, suggesting that two AI at 12h intervals and closer to the time of ovulation may be detrimental. Overall, these data suggest that for sows injected with PG600 at weaning and receiving two AI, breeding at 0 h and 24h after onset of estrus is recommended.     

Effect of PGE2 on uterine contractility and tone in mares

A technique for transvaginal, ultrasound-guided intrauterine injection was developed. After preliminary study using different approaches, the procedure was successful in 24 of 25 (96%) mares, based on detecting fluid in the uterine lumen during and after the injection. The technique was used to study the effect of PGE2, reportedly produced by the embryonic vesicle, on uterine contractility on Day 12 (Day 0 = ovulation). Uterine contractility was scored (1 = minimal, 4 = maximal) every 10 min for 1 h and every 30 min for the next hour by a continuous 1-min ultrasound examination of a longitudinal section of the uterine body without knowledge of group. In Experiment 1, the main effect of group (1-mL vehicle, n = 6; 0.25 microgram PGE2, n = 7) tended to be significant (P < 0.09), and the effect of time was significant (P < 0.008). The mean score was higher for the PGE2 group (2.0 +/- 0.1) than for the vehicle group (1.7 +/- 0.1). An increase in contractility occurred between 0 and 5 min in the vehicle group (P < 0.0004) and between 0 and 10 min in the PGE2 group (P < 0.04). In Experiment 2, there was a tendency (P < 0.08) for effect of group (control without injection, n = 6; 1-mL vehicle, n = 6; 0.025 microgram PGE2, n = 6). The PGE2 group (2.0 +/- 0.1) was different from the vehicle group (1.6 +/- 0.1) and the control group (1.6 +/- 0.1). An increase in contractility occurred between 0 and 20 min in the PGE2 group, and the changes were not significant in the other groups. However, scores were higher in the PGE2 group before treatment, and there were no significant effects when data were converted to percentage changes. The results for an effect of intrauterine treatment of PGE2 on uterine contractility are considered uncertain because of the transient increase in contractility from vehicle injections in Experiment 1 and the higher score in the PGE2 group before treatment, with no significant differences in percentages in Experiment 2. Indirectly, however, an effect of PGE2 was suggested by a shorter (P < 0.05) period of detectability of intrauterine fluid in the PGE2 groups (21 +/- 31 min) than in the vehicle groups (50 +/- 42 min). The shorter period was attributable to greater dispersion of the fluid as a result of increased contractility. In Experiment 3, PGE2 (10 mg, n = 5) and vehicle (4 mL, n = 5) were given intravenously. In addition to uterine contractility, uterine tone was scored (1 = minimal, 4 = maximal) by transrectal digital compression. The main effect of group was significant (P < 0.03) for uterine contractility score, which increased between 0 and 20 min after PGE2 injection. The time effect and interaction were highly significant (P < 0.0001) for uterine tone score, and tone increased in the PGE2 group between 0 and 20 min after injection. The results indicated that PGE2 should be considered as a potential stimulator of both uterine contractions and uterine tone during the time of embryo mobility in mares.     

Antibodies to egg yolk in blood serum of rabbits and cattle and cervical mucus of cattle inseminated artificially.

Serum titers to egg yolk were induced in 6 rabbits by intravaginal deposition of an egg-yolk citrate extender used for artificial insemination of cattle. There was no effect of the low serum titers to egg yolk on fertility of the inseminated rabbits. Titers to egg-yolk semen extender were found in 3% of 59 cows of normal fertility compared to 29% of 14 repeat breeder cows of low fertility, all previously inseminated with semen diluted with egg yolk-citrate extender. Four of 6 cervical mucus samples (67%) from the repeat breeder cows had high titers to egg yolk, but only one also had a positive titer in blood serum.     

Effects of day of estrous cycle, time of day, luteolysis, and embryo on uterine contractility in mares

One-minute continuous ultrasonic scans of longitudinal sections of the uterine body were videotaped, and contractility scores (1 to 5, minimal to maximal contractility) were assigned without knowledge of mare identity, day of the estrous cycle or pregnancy status. Contractility was assessed, and plasma progesterone concentrations were determined for each of 3 daily examinations (at 0800, 1600 and 2400 hours) from Day 9 to Day 19 (Day 0 = day of ovulation). For both the nonbred (n=11) and pregnant (n=11) mares, there was no effect of hour of scan on the extent of uterine contractility. When data for the nonbred mares were normalized to the onset of luteolysis (defined for each mare as the first >/=25% decrease in plasma progesterone concentrations between successive samples), there was an abrupt increase (P<0.05) in contractility 24 hours prior to the onset of luteolysis. Contractility was also assessed daily in 20 nonbred and 27 pregnant mares from Day 0 to Day 17. For the nonbred mares, a biphasic profile in contractility occurred during the estrous cycle as indicated by the following significant changes: a decrease between Days 0 and 2, an increase between Days 2 and 4, a plateau between Days 4 and 7, a decrease between Days 7 and 11, an increase between Days 11 and 13, and a decrease between Days 14 and 16. For pregnant mares, contractility increased (P<0.05) prior to the late-diestrous increase for nonbred mares. In addition, a significant reduction in contractility was detected on Day 5 in these mares compared with that in the nonbred mares. Contractility in the uterine body in 7 mares was assessed every 5 minutes after departure of the embryonic vesicle from the uterine body. Levels of contractility in the uterine body were lower (P<0.05) 55 minutes after the vesicle had exited the body than 相似文献   

The influence of time of insemination relative to time of ovulation on farrowing frequency and litter size in sows, as investigated by ultrasonography

The objective of this experiment was to identify the optimal time of insemination relative to the time of ovulation, based on ultrasonographic detection of embryonic survival at 10 days after ovulation, number of sows farrowing, and litter size. Furthermore, the possible value of the interval from weaning to onset of estrus for prediction of the time of ovulation was examined. Crossbred sows (n = 143) that had farrowed 2 to 9 litters were weaned (Day 0) and observed for estrus every 8 h from Day 3 until end of estrus. Ultrasonography was performed every 6 h, from 12 h after onset of estrus until ovulation had been observed. The sows were inseminated once at various time intervals from ovulation. At Day 16, 25 of the sows were slaughtered and their uteri were flushed for embryos. In the remaining sows, the number of viable and dead piglets and mummified fetuses per sow was recorded at farrowing, with the sum of the 3 constituting the total number of piglets born per sow. The highest number of embryos recovered per sow was found after insemination during the interval from 24 h before to 4 h after ovulation. The lowest frequency of non-pregnant sows and the highest total number of piglets born per sow were found after insemination from 28 h before to 4 h after ovulation. Consequently, the optimal time for insemination was found to be in the interval 28 h before to 4 h after ovulation. The interval from weaning to onset of estrus and from onset of estrus to ovulation were negatively correlated, allowing a rough prediction of the time of ovulation from the interval from weaning to onset of estrus.     

The influence of estradiol and progesterone on the concentrations of uterine oxytocin receptors and plasma PGFM in response to oxytocin in ovariectomized gilts

Peripubertal gilts (n = 25) were treated with corn oil (CO) or ovarian steroids, one month following an ovariectomy. The first day of treatment was assigned as the first day of the experiment. The gilts received: Group (Gr) I (n = 4)--CO (2 mL x day(-1) from 1st to 12th day), Gr II (n = 4) and Gr III (n = 4)--progesterone (P4; 10 to 100 mg x day(-1) from 1st to 12th day), Gr IV (n = 5)--estradiol benzoate (EB; 400 microg x day(-1) from 1st to 3rd day), Gr V (n = 4) and Gr VI (n = 4)--EB + P4 (EB 400 microg x day(-1) from 1st to 3rd day, 20 microg x day(-1) at 6th and 9th day, 50 microg at 12th day plus P4 10 to 100 mg from 4th to 15th day). All gilts were injected with oxytocin (OT; 20 IU; i.v.) on the following days of the experiment: 13th (Gr I and Gr II), 15th (Gr III and Gr IV), 16th (Gr V) and 18th (Gr VI). Concentrations of the PGF2alpha metabolite--PGFM were determined in blood samples, collected from 30 min before to 120 min after OT injection. Baseline PGFM concentrations (30 min before OT) differed among treatment groups and were the highest in Gr V and Gr VI (P < 0.01 vs. other groups). The magnitude of the PGFM response to OT increased only in four of the five gilts of Gr IV and in three of the four gilts of Gr VI, and it was higher (P = 0.009) in Gr VI than in Gr IV. In the remaining groups, PGFM concentrations did not increase above the baseline in response to OT. The day after OT injection, oxytocin receptors (OTR) were found in the uterine tissues of all animals studied. The lowest OTR concentrations were in Gr I--75.5 +/- 11.2 fmol x mg protein(-1) and the highest in Gr IV--712.9 +/- 86.7 fmol x mg protein(-1); (P < 0.05 vs. other groups). The values of K of OTR differed among groups (P < 0.001) and ranged from 1.62 +/- 0.44 nM in Gr I to 12. 08 +/- 1.9 nM in Gr VI. A positive correlation (r = 0.54; P < 0.01) between plasma E2 and uterine OTR concentrations was observed. In conclusion, E2 and P4 are involved in both PGF2 synthesis/secretion and OTR formation, however, full PGF response to OT does not develop before puberty. Estrogens are evident stimulators of uterine OTR synthesis ingilts.     

Synchronization of ovulation and fertility in weaned sows treated with intravaginal triptorelin is influenced by timing of administration and follicle size

A 100 μg dose of triptorelin was tested for synchronizing ovulation in sows. In Experiment 1, conducted in April through June, sows (n = 125) were assigned to Control (untreated), TG-96 (Triptorelin Gel (TG) given intravaginally at 96 h post-weaning), or TG-E (given intravaginally at estrus). To optimize AI timing, sows were inseminated at 2 and 26 h after estrus for Control and TG-E and at 8 and 32 h following TG-96. Ovulation by 48 h post-treatment tended to be affected by treatment (P = 0.08) and more (P < 0.05) TG-96 sows ovulated (57.9%) compared to Controls (34.2%), but TG-E (45.1%) did not differ (P > 0.10). Duration of estrus was reduced (P < 0.005) in TG-96 (51 h) and TG-E (58 h) compared to Controls (65 h). There was no treatment effect on farrowing rate (71%) or total born (10.4). Average follicle size <6.5 mm at 96 h after weaning was associated with reduced (P < 0.01) estrus, ovulation and farrowing rate. Experiment 2 was conducted in August through September using 503 weaned sows. The TG-96 treatment reduced duration of estrus (P = 0.03), but treatment did not affect estrus expression, farrowing rate or total pigs born. In conclusion, use of a 100 μg dose of triptorelin intravaginally at 96 h or at estrus advanced ovulation and when used with timed insemination, resulted in similar farrowing rates and litter sizes comparable to sows mated based on estrus. However, ovulation induction and timed AI success may benefit from an approach that ensures sows have adequate follicle development at time of treatment.     

Effect of altering dose of PG600 on reproductive performance responses in prepubertal gilts and weaned sows

This study evaluated the effects of altering dose of PG600 on estrus and ovulation responses in prepubertal gilts and weaned sows. Experiment 1 tested the effects of one (1.0x, 400IU eCG+200IU hCG, n=74), one and a half (1.5x, n=82), or two (2.0x, n=71) doses of PG600 for prepubertal gilts. Estrus (58%) and ovulation (90%) were not affected (P>0.10) by dose. Higher doses increased (P<0.01) numbers of corpora lutea (17, 24, and 25), but not (P>0.10) the proportion of gilts with cysts (26, 36, and 46% for 1.0x, 1.5x, and 2.0x, respectively). Experiment 2 tested the effects of 0x (n=30), 0.5x (n=32), 1.0x (n=29), or 1.5x (n=30) doses of PG600 in weaned sows. Dose did not influence return to estrus (90%, P>0.10). There was an effect of dose (P<0.05) on incidence of cysts (3.4, 1.8, 6.4, and 29.8%, for 0x, 0.5x, 1.0x, and 1.5x doses, respectively). The 0.5x dose increased (P<0.01) farrowing rate (83.2%) compared to 0x (72.1%) and 1.5x (58.6%), but was not different from 1.0x (76.4%). Total pigs born (10.5+/-0.8) did not differ (P>0.10) among treatments. These data suggest that increasing dose of PG600 to 1.5x for gilts increases the number of corpora lutea but does not alter the proportion expressing estrus or ovulating. Reducing dose of PG600 for weaned sows did not alter estrus or ovulation, but the 0.5x dose increased farrowing rate compared to no PG600.     

Effect of naloxone on plasma concentrations of prolactin and LH in lactating sows

Six lactating sows were injected through an indwelling vena cava cannula with naloxone (2.5 mg/kg body weight) on Day 15 post partum. Blood samples were collected through the cannulas at 10-min intervals for 8 h before and 10 h after naloxone administration. Plasma prolactin and LH concentrations were measured by radioimmunoassay. Naloxone caused a marked suppression of plasma prolactin concentrations lasting 4-6 h. LH concentrations were also affected by naloxone: LH rose to reach maximum values 20-50 min after naloxone treatment. Pretreatment values were recorded 200-300 min after the treatment. These results indicate that endogenous opioids are involved in causing the endocrine patterns occurring during lactation, i.e. high prolactin and low LH concentrations.     

Amikacin sulfate in the cat: serum, urine and uterine tissue concentrations

Puerperal metritis and pyometra in non-breeding cats is frequently caused by gram-negative bacteria that are resistant to a variety of antibiotics. Amikacin has been found to be effective against pathogens associated with uterine infections in the mare and the woman, but its efficacy has not been studied in the cat. Serum concentrations of amikacin were determined in healthy adult cats (six male and six female) after administration of 5, 10, and 20 mg/kg body weight of amikacin sulfate, each dose given subcutaneously (s.c.), intramuscularly (i.m.) and intravenously (i.v.) to each of the cats using a repeat treatment design. In a subsequent experiment, the six females were given 10 mg/kg s.c. amikacin and samples of blood, urine and full-thickness uterine wall were taken at 40 and 120 minutes after treatment. Mean serum concentrations of amikacin peaked between 30 and 45 minutes after i.m. injection and between 45 and 60 minutes after s.c. injections. The serum amikacin concentration curves were similar regardless of dose or administration route except for a slightly longer retention time after the 20 mg/kg dose given i.m. and s.c. After s.c. injection of 10 mg/kg, the mean uterine concentration of amikacin at two hours after treatment was 4.1 ug/g; the concurrent mean serum concentration was 18.6 ug/ml.     

Effects of altering the dose and timing of triptorelin when given as an intravaginal gel for advancing and synchronizing ovulation in weaned sows

Previous studies have shown that triptorelin gel (TG) given intravaginally in gel form is effective for advancing the time of ovulation in weaned sows. Three experiments were performed to determine the effects of altering the dose and timing of administration of intravaginal TG for advancing and synchronizing ovulation in weaned sows. In all experiments, estrus was detected twice or three times daily and ultrasound was performed to determine ovulation at 8-hour intervals. In experiment 1, sows (n = 131) received intravaginal gel containing 0 (Placebo), 25, 100, or 200 μg of TG at 96 hours after weaning and sows were inseminated on each day of standing estrus. Wean-to-estrus interval and duration of estrus were correlated (P < 0.0001) with estrus duration longer in TG (P < 0.05) compared with Placebo. More sows ovulated (P < 0.001) by 48 hours after treatment with 200 (81%), 100 (64%), and 25 μg (63%) of TG compared with Placebo (42%). The farrowing rate and total pigs born did not differ (P > 0.10). In experiment 2, sows (n = 126) received 200 μg of TG at 72, 84, or 96 hours after weaning or were untreated (Control-96). Sows receiving TG were inseminated once 24 to 28 hours after treatment. Control-96 sows were inseminated on each day of standing estrus. Wean-to-estrus interval was not affected by treatment, but wean-to-ovulation interval was reduced (P < 0.05) by TG-72 and TG-84 compared with TG-96 and Control-96. More sows ovulated 40 hours after treatment (P < 0.001) with TG-72 (56.5%) and TG-84 (32.2%) compared with TG-96 and Control-96 (13%) and for all TG treatments 48 hours after treatment (64%) compared with Control-96 (34%, P < 0.05). The farrowing rate was lower (P < 0.05) for sows assigned to TG-72 and TG-84 compared with TG-96 and Control-96, whereas the number of liveborn pigs did not differ (P > 0.10). In experiment 3, sows (n = 113) were assigned to receive no treatment (Control), intravaginal gel alone (Placebo), or 200 μg of TG given intravaginally (OvuGel) at 96 hours after weaning. Wean-to-estrus interval did not differ, but the duration of estrus tended (P < 0.10) to be reduced with OvuGel compared with the other treatments. More sows ovulated (P < 0.001) by 48 hours after OvuGel treatment (79.1%) compared with Control (46.4%) and Placebo (37.9%) and by 56 hours (P < 0.05). The farrowing rate and the number of liveborn pigs did not differ among treatments. The results of these studies indicate that 200 μg of TG given intravaginally at 96 hours after weaning (OvuGel) synchronizes ovulation and results in fertility similar to Controls.     

Effect of mouse epidermal growth factor on plasma concentrations of FSH, LH and progesterone and on oestrus, ovulation and ovulation rate in merino ewes

The 24 h i.v. infusion of Merino ewes with 60 or 100 microgram mouse epidermal growth factor (EGF)/kg body weight on Days 4, 9 or 14 of the oestrous cycle decreased the strength of wool attachment and caused marked changes in subsequent reproductive performance. In ovaries removed 2 days after EGF treatment all follicles greater than or equal to 0.6 mm diameter were atretic. After 7 days either a normal pattern of atresia or no atresia was evident while after 12 days the pattern of follicular atresia was similar to that in controls. Irrespective of stage of cycle EGF caused dose-dependent increases in plasma FSH concentrations that persisted for up to 14 days. Changes in plasma LH concentrations were generally similar after infusion on Days 4 and 14, but were smaller and shorter-lived after infusion on Day 9. Irrespective of dose, the infusion of EGF on Days 4 and 14 caused immediate luteolysis then the formation of a luteinized follicle in many ewes. Most ewes treated on Day 4 returned to oestrus between Days 17 and 21 with the same ovulation rate (1.3) as the controls. Of those infused on Day 14 oestrus occurred about a cycle length later than expected and their ovulation rate then (1.9) was also similar to that of the controls (1.7). Luteal function was not affected in ewes infused on Day 9, and most returned to oestrus between Days 17 and 20 with an ovulation rate of 3.2. Fertile rams were not placed with the ewes until after the differences in ovulation rate had been observed. Mating occurred generally 2-4 weeks after treatment, and there were no differences between EGF-treated and control ewes in fertility or fecundity. The results are interpreted as indicating that mouse EGF induces ovarian follicular atresia but has differential effects on luteal function according to the stage of the oestrous cycle at which it is given. As a consequence of these two effects, which lead to differential changes in gonadotrophin secretion, ovarian function may be temporarily impaired, little affected or improved.     

Influence of serum luteinising hormone concentrations on ovulation,conception, and early pregnancy loss in polycystic ovary syndrome.

Women with the polycystic ovary syndrome do not respond well to treatment with luteinising hormone releasing hormone. To determine whether this might be due to an underlying endocrine disturbance basal concentrations of luteinising hormone were measured in 54 infertile women treated with pulsatile luteinising hormone releasing hormone and concentrations at the time of maximum follicular growth were measured in 23 of the patients. Forty one patients ovulated. Forty one patients ovulated and 27 conceived, but nine pregnancies terminated within four weeks after ovulation. Basal luteinising hormone concentrations were significantly lower in those who conceived (12.4 (range 1.3-29.0) IU/l) than in those who did not (19.0 (3.5-50.0) IU/l) and in those whose pregnancy progressed (9.6 (1.3-29.0) IU/l) than in those with early loss of pregnancy (17.9 (7.0-29.0) IU/l). Concentrations at the time of maximum follicular growth were significantly lower in women who ovulated (9.4 (2.9-35.4) IU/l) than in those who did not (29.0 (7.0-50.0) IU/l) and in those who conceived (6.2 (2.9-8.5) IU/l) than in those who did not (17.9 (4.0-50.0) IU/l). These results indicate that high concentrations of luteinising hormone during the follicular phase in women with polycystic ovaries have a deleterious effect on rates of conception and may be a causal factor in early pregnancy loss.     

Nitroprusside stimulates contractility and the synthesis of 14C-acetylated PAF-like substances in estrogen primed-mouse uterine horns.

We investigated the effect of sodium nitroprusside (SNP), a donor of nitric oxide, on the formation of platelet-activating factor (PAF) and uterine contractility in mouse uterine horns from mice treated with estrogen. Because the major pathway of PAF synthesis is the remodeling pathway in uterine tissue, we evaluated the incorporation of 14C-acetate into PAF-like molecules. Our results showed that SNP (100-300 mumol/L) caused a transient increase in the synthesis of PAF, which remained cell-associated. The addition of SNP (100-300 mumol/L) to a mouse uterine horn in an isolated organ bath preparation evoked a transient increase in contractility, which was inhibited by hemoglobin (2 micrograms/mL), a nitric oxide scavenger, but not by methylene blue (10 mumol/L), a guanylate cyclase inhibitor. The pharmacological characteristics of the contractions evoked by SNP resembled those evoked after mast cell activation, in that they were blocked by ritodrine (a beta 2 adrenergic agonist, 0.1 mumol/L); indomethacin (a cyclooxygenase inhibitor, 10 mumol/L); ketotifen (a mast cell stabilizer, 1.0 mumol/L); cromolyn sodium (a mast cell stabilizer, 100 mumol/L); pyrilamine (an H1 antagonist, 10 mumol/L); and ketanserine (5HT2 antagonist, 0.1 mumol/L). These data demonstrate that nitric oxide generated from SNP stimulated the synthesis of PAF and evoked contractility in uterine horns from mice treated with estrogen. This result suggests the possibility that these tissue conditions might be favorable for the generation of peroxynitrites, possible mediators of both effects. It is also shown that the contractility evoked by the addition of SNP was not due to production of PAF, because its antagonist, WEB 2086 (10-30 mumol/L, a concentration that blocked contractions evoked by PAF 1 nmol/L), had no effect on the SNP-evoked contractions.