Fertility studies in young and mature merino ewes 1. Cervical mucus production

The production of cervical mucus was determined at six-hour intervals during oestrus in young (1 (1 2 ) years old) and mature (3 to 7 years old) Merino ewes. The length of the cervix and the length of the cervical epithelium was also measured in similar ewes. Total weight of mucus secreted as well as secretion rate per unit of time was greater in mature ewes than young ewes. Production was maximal during the first six hours of oestrus and declined considerably after 12 hours in young and 18 hours in mature ewes. The cervix was significantly shorter in young ewes while the length of the cervical epithelium was not significantly different in young and mature. It was concluded that lower mucus production in young ewes is not related to lower mucus output per unit area.     

Oestrous cycle dynamics in peri-pubertal and mature Javanese thin-tail sheep

Duration of oestrus, time of ovulation and hormone profiles for progesterone and LH in prepubertal, pubertal and mature Javanese thin-tail sheep were studied at synchronized oestrus following progestagen-PMSG treatment and at the first natural oestrus after synchronization.The ewe lambs responded to progestagen-PMSG treatment by showing earlier onset of oestrus and an earlier and higher peak of LH concentration than mature ewes. For pre-pubertal, pubertal and mature ewes the mean LH peaks were 49.9, 43.9 and 37.9 ng/ml (P>0.05) at mean intervals of 7.5, 8.4 and 16.5 h (P < 0.05), respectively, after onset of oestrus. Duration of oestrus was 41.2 h in pubertal lambs and averaged 37.5 h in the other two groups (P>0.05). Except in one mature ewe, ovulation occurred between 24 and 36 h after onset of oestrus and the majority ovulated at around the end of oestrus. The corpora lutea developed normally, as indicated by plasma-progesterone changes. The patterns of plasma-progesterone changes were similar in all three groups, though the concentrations were lower in the ewe lambs.At the first natural oestrus after synchronization, mature ewes showed longer (P>0.05) oestrus (31.5 vs. 24.3 h), longer time interval from onset of oestrus to the LH peak (16.0 vs. 12.0 h) and from the LH peak to ovulation (21.0 vs. 19.6 h) than peri-pubertal lambs. Six of eight pre-pubertal lambs did not ovulate at their first natural oestrus, resulting in a conception rate of 11% for that group, while in pubertal lambs and mature ewes conception rates were 70% and 100%, respectively.     

Impact of ACTH during oestrus on the ultrastructure of the spermatozoa and their environment in the tubal reservoir of the postovulatory sow

This study investigated whether injections of synthetic ACTH (simulating short-term stress) in sows during standing oestrus have a negative effect on spermatozoa and the local intraluminal environment in the utero-tubal junction (UTJ) and isthmus. Seven of the 14 sows were given ACTH through a jugular catheter every 2 h from the onset of standing oestrus until the sow ovulated (ACTH-group), while the other seven sows were given NaCl solution (C-group). All sows were artificially inseminated before ovulation. Six hours after ovulation (detected with transrectal ultrasonography) the sows were anaesthetised, the right oviduct was fixed in toto by vascular perfusion with glutaraldehyde, and the UTJ and specimens from the isthmus were prepared for scanning electron microscopy (SEM). SEM revealed that a seemingly viable population of spermatozoa remained in the UTJ 6 h after ovulation. A majority of sows in the ACTH-group had moderately to exaggerated amounts of mucus in the intraluminal environment of the sperm reservoir. In conclusion, stress simulated by exogenous ACTH in sows may alter the intraluminal environment of the sperm reservoir.     

Ovarian programming of gamete progression and maturation in the female genital tract

Whilst the rate of displacement and migration of sperm cells in the female reproductive tract of rodents, farm animals and humans has attracted attention for at least 50 years, the overriding purpose of sperm transport has not always been kept in focus. This report is concerned with spermatozoa that can penetrate the egg investments and promote formation of a zygote, judgements involving a surgical approach and subsequent phase-contrast microscopy. A minimum period of 6–8 hours was required for such spermatozoa to be established in the oviducts in sheep and cows mated at the onset of oestrus. Sperm were then arrested in the caudal 12 cm of the isthmus for 17–18 hours or more until just before the moment of ovulation, when they were activated and displaced onwards to the site of fertilization at the ampullary-isthmic junction. The time-scale of these events differs in pigs as a result of the intra-uterine site of ejaculation and the 40-hour interval between the onset of oestrus and ovulation, but the pre-ovulatory sequestering of viable spermatozoa in the caudal tip of the oviduct is conspicuous for 36 hours or more. This function of the oviduct appears to be under local control from ovarian follicular hormones and, as judged by sperm motility and membranous changes, so does the process of capacitation. Completion of capacitation is interpreted as a peri-ovulatory event.     

Human sperm-cervical mucus interaction and the ability of spermatozoa to fuse with zona-free hamster oocytes

Samples of semen and cervical mucus were provided by 18 couples. Cervical mucus was obtained for each day possible and stored at 4 degrees C until all the samples were collected. Flat capillary tubes were loaded with the mucous samples and spermatozoa from the husband's semen sample were allowed to migrate through the cervical mucus (3 cm column) into culture medium. The spermatozoa recovered after migration through cervical mucus were assayed in vitro with zona-free hamster oocytes. Control experiments were carried out using spermatozoa from the same semen sample but prepared by the swimming-up technique. Altogether, 557 eggs in the control group and 1236 eggs in the experimental group were analysed, and the results demonstrated that the % of sperm penetration, the mean number of sperm decondensations per penetrated egg and the mean number of spermatozoa adhering per egg all had higher values (P less than 0.05) for the control samples than for the experimental samples. We suggest that cervical mucus modifies human spermatozoa, as measured by their interaction with zona-free hamster oocytes.     

Reproductive response of ewes synchronized with different lengths of MGA treatments in intrauterine insemination program

A total of 415 fat tailed ewes were randomly assigned to two groups to assess the effect of duration of melengestrol acetate (MGA) (9 versus 12d) administration on reproductive parameters associated with laparoscopic artificial insemination. At the end of MGA treatment, ewes in each group were subdivided and inseminated with one of two different insemination doses (10×10(7) or 20×10(7) sperm per 0.5 ml insemination dose) of fresh diluted semen. Inseminations were carried out 11-18 h after first detected estrus. Ewes were screened for their return to oestrus from 10 to 21 days post AI and inseminated at their returned oestrus. Pregnancy diagnosis was done from approximately 55 days after insemination in both synchronized and return estrus. For short (9-day) and long (12-day) term MGA treated groups, estrus rates were 62% versus 89% (P<0.0001), respectively. Ewes (n=115) that returned to estrus were inseminated (7-11h after estrus detection) with fresh diluted semen at different doses (20×10(7) or 40×10(7) or 60×10(7) sperm per 0.5 ml insemination dose). Pregnancy rates were 41% and 44% for short term and long term MGA treated ewes, respectively. Pregnancy rate of ewes which returned to oestrus was 53.4%. There was a significant (P<0.05) increase in pregnancy rates (38-52% for 11-16 h; 63% for 17-18 h) when insemination was held at 17-18 h after first detected estrus following MGA treatments. Pregnancy rates were found to be similar in ewes inseminated with 10×10(7) (36%) or 20×10(7) (47%) motile spermatozoa at first AI, and 20×10(7) (44%) or 40×10(7) (59%) or 60×10(7)(48%) at second AI. It was concluded that short term MGA treated ewes were recorded with lower estrus rates but was similar to pregnancy rates with long term MGA treatment. Acceptable pregnancy rates were achieved in MGA induced estrus when insemination is conducted at 17-18 h after estrus onset and with 20×10(7) sperm per insemination dose.     

The effect of oestrogen administered during the progestational phase of the cycle on transport of spermatozoa in ewes.

Two split-plot factorial experiments are described, the first with 72 entire cyclic ewes and the second with 80. The pattern of transport of spermatozoa through the reproductive tract was studied, following treatments with progestagen and oestrogen or with oestrogen alone during 2 weeks preceding insemination. A daily dose of 25 mug oestradiol-17 beta administered to ewes for 14 days preceeding oestrus had a deleterious effect on the passage of spermatozoa through the cervix into the uterus within the first 2 hr after insemination. The numbers of spermatozoa recoverable from the cranial region of the cervix 2 hr after insemination appeared to be related to the numbers in the oviducts at 24 hr. These numbers were related to fertility data from an earlier experiment using similar treatments. The data for log numbers of spermatozoa recoverable from the cervix formed a near-normal distribution and so were suitable for formal statistical analysis. There was an interaction between progestagen and oestrogen influence before mating on the pattern of sperm transport through the cervix.     

Comparison of seminal quality in Holstein bulls as yearlings and as mature sires

Semen quality was compared in 5 Holstein bulls from samples collected as young sires (yearlings) and again as mature bulls after a mean interval of 1,265 d. At both sampling periods, the semen was examined for ejaculate volume, sperm numbers, post-thaw progressive motility and sperm viability. Sperm viability was assessed on cryopreserved samples with fluorescent SYBR-14 to stain living spermatozoa and propidium iodide (PI) to identify dead spermatozoa. The fluorescent populations of stained spermatozoa were quantified by flow cytometry. The percentages of living spermatozoa for the individual bulls, as determined by green fluorescence of SYBR-14, ranged from 44 +/- 3.1 to 54 +/- 0.3 for yearlings, and from 38 +/- 1.5 to 55 +/- 1.0 for mature sires. No differences in sperm viability were found between samples taken from yearling bulls and those of mature bulls. The percentage of spermatozoa stained with SYBR-14 was negatively correlated (r = -0.97; P = 0.0001) with the percentage of dead spermatozoa as indicated by PI staining. Comparisons of identical samples run on 2 different flow cytometers indicated that either flow instrument could be used to assess sperm viability. Although the individual bulls differed (P < 0.05) in ejaculate volume and sperm numbers as yearlings, they did not differ in these parameters as mature bulls. The average number of spermatozoa per ejaculate changed as a result of maturation, increasing from 6.2 +/- 1.0 to 10.7 +/- 1.1 x 10(9). Aging was significantly correlated with ejaculate volume (r = 0.76; P = 0.01) but not with the total number of spermatozoa per ejaculate (r = 0.51; P = 0.13). The maturational changes that occurred in the 5 bulls were minimal with the exception of the increased volume of the ejaculate and the number of spermatozoa per ejaculate.     

Impact of ACTH administration on the oviductal sperm reservoir in sows: the local endocrine environment and distribution of spermatozoa

The objective of the study was to investigate if short-term stress in sows (simulated by injections of synthetic adrenocorticotrophic hormone (ACTH)) during standing oestrus had a negative effect on the local environment in the utero-tubal junction (UTJ) and isthmus and the distribution of spermatozoa in these segments. Fourteen sows were monitored for ovulation using ultrasonography in two consecutive oestruses. The sows were fitted with jugular catheters and, from onset of the second oestrus, blood samples were collected every second hour. In the 2nd oestrus, seven sows were given ACTH every second hour, from the onset of standing oestrus until the sow ovulated (ACTH-group), whereas the other seven sows remained as controls (C-group) and were given NaCl solution. The sows were artificially inseminated 16-18 h before expected ovulation. Six hours after ovulation the sows were anaesthetised, and blood samples were repeatedly taken from veins draining the uterus and the UTJ-isthmus, respectively. This oviduct was thereafter removed and divided in four adjacent sections consisting of: (i) the UTJ, (ii) the first, and (iii) the second isthmus segment prior to (iv), the ampullary-isthmic junction (AIJ) and the ampulla. The three first-mentioned segments were flushed to retrieve spermatozoa, whereas the last one was flushed to collect oocytes/ova. The number of spermatozoa attached to the zona pellucida was counted. The concentrations of cortisol in jugular blood of the ACTH-group sows during the time of ACTH-injections were significantly higher than of the C-group sows (p<0.05), as were the levels of progesterone (p<0.001). Progesterone and cortisol concentrations measured in the blood samples draining the UTJ-isthmic region 6 h after ovulation did not significantly differ between the groups, but the C-group displayed significantly higher concentrations of progesterone in the UTJ-isthmic region compared with the levels measured in parallel samples taken of jugular blood (p<0.01). The C-group, but not the ACTH-group, also displayed a significant elevation in progesterone concentration 6h after ovulation compared with the basal levels before ovulation (p<0.01). Numbers of retrieved spermatozoa were not significantly different between the C-group and the ACTH-group. However, there was a tendency for a larger number of spermatozoa among sows in the ACTH-group, especially in the isthmic segment adjacent to the AIJ. In conclusion, simulated stress induced by injections of ACTH during standing oestrus results in elevated concentrations of progesterone before ovulation and may interfere with the rise of progesterone after ovulation. However, ACTH-injections appeared to augment transport of spermatozoa through the female genital tract of pigs.     

Pulsatile release of oxytocin into the circulation of the ewe during oestrus, mating and the early luteal phase

Two experiments were designed to investigate release patterns of oxytocin into plasma during oestrus and the early luteal phase. In Exp. 1, blood samples were collected from 5 ewes every 30 min for 10 h during 6 days around oestrus and the early luteal phase. During oestrus concentrations of oxytocin were generally low (1.27 +/- 0.54 pg/ml; mean +/- s.d.) but with occasional pulses up to 6 pg/ml. By Day 5 mean basal concentrations had risen to 4.5 +/- 2.1 pg/ml with a fluctuating release pattern. In Exp. 2, a method was developed for continuous blood sampling from conscious, unrestrained ewes. On the predicted day of oestrus following an untreated oestrous cycle, 8-ml blood samples were collected every minute for two 35-min periods (8 ewes: 16 sampling periods). For 6 ewes a ram was introduced to the pen for part of this time, and resulting behaviour was recorded. Additional blood samples were assayed for LH and progesterone to determine the stage of the cycle. Overall mean oxytocin concentrations ranged from 1.5 +/- 0.53 to 6.8 +/- 5.25 pg/ml in different animals. Ewes which were both in oestrus and exposed to the ram showed a pulsatile oxytocin release pattern consisting of low baseline concentrations with short-duration pulses superimposed (duration 1-4 min; amplitude 2.5-31.7 pg/ml; frequency 3.18/h). Coitus was not temporally associated with pulsatile release. However, the importance of the presence of the ram was indicated by total separation of 2 oestrous ewes from the ram until after experimentation. In these animals only 1 pulse of oxytocin was detected in 2.7 h of sampling. It is concluded that, although mean oxytocin concentrations at oestrus were low, short duration pulses were released into the plasma at this time. This effect may be dependent on the presence of a ram.     

Oestrogen and progesterone concentrations in plasma and oviductal tissue of ewes exhibiting a natural or induced oestrus

Synchronisation of oestrus in Karagouniki ewes by administration of the standard dose of progesterone results in lower fertility than observed when these ewes ovulate naturally. This suggests that the optimum dose of progesterone may be breed dependent. The exogenous progesterone may perturb the concentrations of oestradiol-17beta and progesterone in blood plasma and the oviductal wall. This possibility was investigated using Karagouniki ewes allocated at random to three treatments (n=4 per treatment). Ewes were allowed to exhibit natural oestrus (N) or oestrus was synchronised by administration of 250 mg (LP) or 375 mg (HP) progesterone (subcutaneous implants) followed by PMSG at 8 mg/kg live weight i.m. 14 days later. Oestrus was observed using teaser rams. Blood samples were collected for plasma oestradiol-17beta and progesterone assay from the onset to the end of oestrus at 2 h intervals. The uterus of each ewe was recovered at the end of oestrus and samples of the oviductal wall were taken from both oviducts and prepared, separately, for progesterone and oestradiol-17beta assay. Statistical analysis was performed using univariate analysis of variance. Plasma oestradiol-17beta concentrations from the onset to the end of oestrus were highest for N ewes and lowest for HP ewes with the values for LP ewes occupying an intermediate position. The differences were significant (P<0.05) between HP and the other two treatments from 4 to 12 h after the onset of oestrus and then between all treatments until the end of oestrus. Plasma progesterone levels were similar and fairly constant from the onset to the end of oestrus for N and LP. The plasma progesterone levels for HP were significantly (P<0.05) higher than for the other two treatments throughout oestrus. In oviductal wall samples, the oestradiol-17beta concentration was significantly (P<0.05) higher for N ewes than for synchronised ewes and the levels were similar for LP and HP ewes. The concentration of oestradiol-17beta differed (P<0.05) between right and left oviducts for N ewes but not for ewes of either of the synchronised oestrus treatments. Progesterone concentrations in oviductal wall samples were highest (P<0.05) for HP ewes and the values for N and LP ewes were similar. The concentration of progesterone did not differ between right and left oviductal wall samples within treatments. It was concluded that the higher dose of exogenous progesterone perturbed the levels of oestradiol-17beta and progesterone in blood plasma and the oviductal wall, and this could explain the lower levels of fertility (relative to naturally occurring oestrus) observed when this protocol is used for Karagouniki ewes in practice.     

Fertility and sperm transport in Merino ewes at the first oestrus following embryonic death.

The embryos of ewes were killed with colchicine on Day 17 of gestation and the ewes were mated at the subsequent oestrus. Fertility was reduced at this mating, and fewer spermatozoa were found in the uterus and oviducts than in control animals. The total number of spermatozoa in the cervix and their distribution between the lumen and walls of the cervix were not altered, but the linear distribution along the cervical walls was changed. The density of the reamining spermatozoa in the control animals after flushing the cervix showed a progressive decrease from the posterior to the anterior segments. This did not occur in the untreated ewes. It seems likely that impaired sperm transport contributed to the lowered fertility.     

8-hour rhythm of ovarian progesterone secretion in ewe

Pulse character of hormones secretion in the hypothalamus-pituitary-gonads system is a necessary condition of physiological regulation of reproduction. At the same time, the rhythms of ovarian hormones secretion have not been adequately explored. The researches study mainly three sexually mature ewes. The stages of oestrus cycle were determined on behavioral reactions of females in the presence of ram. Blood samples from jugular vein were collected hourly over 24-hour period during follicular (15-16 days), early (3-4 days) and middle (7-9 days) luteal phase of oestrus cycle, pregnancy (40-105 days) and lactation (30-45 days). 27 experiments were performed. Plasma progesterone was determined by enzyme-immunoassay method. There was no diurnal rhythm of ovarian progesterone secretion in ewes. During early and middle luteal phase of oestrus cycle and lactation, an 8-hour rhythm of progesterone secretion was detected. Follicular phase of oestrus cycle and pregnancy were characterized by irregular rises of fluctuations of progesterone level. It seems that the 8-hour rhythm of progesterone secretion during luteal phase and lactation is controlled by action of intraovarian generator of ultradian rhythms.     

Controlled sheep breeding: update 1980-1985

This contribution to the Symposium concerns four topics which have been addressed in our laboratory over the past five years. First, the responses to a controlled light environment of Merino ewes and rams have been compared with those of two British breeds. The endocrinological patterns were similar in all breeds but cyclic ovarian activity and ram libido were different. While showing a degree of entrainment to photoperiod, the breeding patterns were much less rigidly controlled in the Merinos than in the others. Second, the effectiveness of establishment of a cervical reservoir of spermatozoa, in ewes in which oestrus and ovulation have been controlled, has been re-examined. This is highly dependent on the time of insemination relative to that of the release of LH. Maximum numbers are found when ewes are inseminated shortly after the LH peak, i.e. some 6-10 h after the onset of oestrus. Third, the quantitative and temporal endocrinological and behavioural events following standard, progestagen-PMSG treatment have been quantified. Contrary to earlier expressed beliefs, these events are remarkably predictable provided an intensive system of mating or detection of oestrus is used. The onset of oestrus in treated anoestrous crossbred ewes has a normal distribution, with a range of 24 h, centred around a mean of 33 h after withdrawal of a 30 mg Cronolone intravaginal sponge and injection of 500 i.u. PMSG. This period of time is dose-dependent. The LH peak occurs 4.5 +/- 0.7 h later and the times of onset of oestrus and of LH release are highly correlated (r = 0.93). Ovulation is some 24 h later again. Fourth, differences in the response of ewes to different batches of PMSG have been defined. While the three commercial preparations studied regularly induced ovulation in anoestrous ewes at doses of 250 i.u. and above, the quantitative responses varied greatly. One preparation would not induce multiple ovulation, even at high doses. There are differences in steroidogenesis and in pregnancy rates, associated with dose of PMSG and the consequent ovulation rate: the ideal would be for every ewe to shed two or three ova. A higher ovulation rate is acceptable, as early embryonic mortality generally reduces the litter size. This is particularly important in deep anoestrus. However, this does not solve the problem of breeding in early lactation.     

The effects of nutrition on the reproductive performance of Finn x Dorset ewes. II. Post-partum ovarian activity, conception and the plasma concentration of progesterone and LH.

After lambing forty-five ewes were allocated to three groups, two of sixteen and one of thirteen ewes. The lambs of the two groups of sixteen ewes were weaned on Day 1 after lambing and the ewes were fed a diet of 100% (Group H) or 50% (Group R) of maintenance energy requirements. The thirteen ewes in the third group (Group L) suckled twin lambs and were fed freely. During the first 3 weeks after lambing, oestrus was observed for 11/16 (Group H) and 8/16 (Group R) ewes; of the ewes which had shown oestrus in the two groups, ovulation occurred in 5/8 and 5/7 respectively. Only 1/13 Group-L ewes showed oestrus and ovulated during the same period. The mean plasma concentrations of progesterone and LH were unaffected by the treatments and were around 0-4 and 1-5 ng/ml, respectively. Restricted feeding had no effect on oestrus, ovulation or the hormone levels during the oestrus cycle following synchronization. The onset of oestrus and the start of the preovulatory discharge of LH were 3 and 6 hr later, respectively, in the lactating ewes (Group L) than in those in Groups H and R. Ewes in Group L also had a higher ovulation rate, 2-8 +/- 0-2 versus 2-1 +/- 0-2 (P less than 0-05). Restricted feeding reduced the number of ewes lambing; only 1/11 ewes in Group R, considered to have conceived because of the presence of high progesterone levels 17 days after mating, subsequently lambed compared with 6/12 in Group H and 5/9 in Group L.     

Effects of scrotal insulation and unilateral vasoligation on ejaculate characteristics and sperm reserves in the bull

Ten mature Bos indicus cross bulls were unilaterally vasoligated and the scrotum was insulated for 48 hours in six of the animals (treated). Semen was collected at 2-day intervals by electroejaculation, until the reproductive tract was recovered at slaughter either 14, 20 or 26 days after treatment. Decapitate spermatozoa and spermatozoa with proto-plasmic droplets increased significantly (P < 0.01) over pre-treatment values at 12 to 16 and 16 to 20 days, respectively, after insulation. Vasoligation and scrotal insulation had no effect on testicular sperm reserves, but epididymal sperm reserves on the ligated side were significantly (P < 0.05) reduced in the caput and increased in corpus and cauda. Total numbers of spermatozoa recovered from the patent side (epididymal sperm reserves and ejaculatory sperm output) exceeded sperm reserves recovered from the ligated side at 20 (28.2 vs. 23.2 × 10⁹) and 26 days (47.1 vs. 18.7 × 10⁹) after treatment in the insulated bulls. The percentage of decapitate spermatozoa was higher and the proportion of spermatozoa with protoplasmic droplets was consistently lower in the ligated cauda epididymidis. These findings are discussed in relation to likely alterations in epididymal function following scrotal insulation.     

Cervical mucus and oestrous behaviour responses to oestrogens in sheep: Progesterone-oestrogen relationships and role of progesterone pre-treatment

The final dose of progesterone (5, 10, 20 mg) and time to oestrogen injection relative to the final dose of progesterone (24–72 h) had no significant effect on the production of cervical mucus measured 24 h after the injection of 30 μg oestradiol benzoate (ODB). However, there were significant effects on the behavioural oestrous responses (time from injection of oestrogen to onset of oestrus and duration of oestrus). Time to onset of oestrus increased from 18 to 27.8 h with increasing dose of progesterone (P < 0.001) and decreased from 24.8 to 20 h with increasing time to oestrogen injection (P < 0.05). Conversely, the duration of oestrus decreased from 36.2 to 23.8 h with increasing dose of progesterone (P < 0.001) and increased from 29 to 39 h with increasing time to oestrogen injection (P < 0.01).Ovariectomized ewes became refractory to ODB as measured by the cervical mucus response after the fifth sequential daily injection of 20 μg oestradiol benzoate. Progesterone priming was not required to restore subsequent sensitivity to oestrogen treatment. However, there was a positive linear relationship between length of recovery period and level of response to subsequent treatment.It was concluded that: (1) progesterone pre-treatment or priming is not necessary in the cervical mucus bioassay in ovariectomized ewes; and (2) a period of 8–16 days is needed between assays for normal sensitivity to be regained.     

Ewe breed differences in fertility after cervical AI with frozen-thawed semen and associated differences in sperm penetration and physicochemical properties of cervical mucus

The objective of this study was to examine sperm penetration through cervical mucus and associated physicochemical properties of cervical mucus from Belclare and Suffolk ewes - two breeds with divergent pregnancy rate following cervical AI using frozen-thawed semen. In Experiment 1, sperm penetration through cervical mucus was assessed in 15 Belclare and 15 Suffolk ewes at 30, 48 and 57h post sponge removal. In Experiment 2, rheological properties of mucus from 17 Belclare and 19 Suffolk ewes at 48 and 57h post sponge removal were determined. In Experiment 3, 20 Belclare and 20 Suffolk ewes were used to assess mucus ferning and pH collected at 42, 48, 57 and 65h post sponge removal. In Experiment 1, a higher number of sperm penetrated cervical mucus from Belclare ewes at 48h, reflected by a breed by time interaction (P=0.05). In Experiment 2, mucus from Suffolk ewes tended to have higher elastic and complex moduli than that from Belclare ewes (P=0.06) regardless of time of collection. There was no effect of ewe breed on the viscous modulus. In Experiment 3, there was a significant effect of time post sponge removal on ferning (P<0.01), but there was no effect of breed. There was no effect of time or breed on mucus pH. It is concluded that breed differences in the rheological properties of cervical mucus affect the ability of sperm to swim through cervical mucus and this may explain breed differences in fertility observed after cervical AI using frozen-thawed semen.     

Composition lipidique des spermatozoides humains et susceptibilité au stress oxydant avant et après migration dans le mucus cervical

Spermatozoa are particularly susceptible to damage induced by ROS, especially as their plasma membrane contains large amounts of polyunsaturated fatty acids. Mammalian sperm cells develop the capacity to fertilise ova during transport in the male and female reproductive tracts. The nature and quality of the micro-environment of the female reproductive tract are important factors for sperm selection, capacitation and subsequent acrosome reaction.In vitro experiments using capacitating media have shown remodeling of the lipid composition of the sperm membrane during these steps and the same approaches have also shown that a low level of ROS was necessary. The oxidative status of the female genital tract is therefore certainly of primary importance for the physiological maturation of the sperm cell. It has been previously reported that an inappropriate oxidative balance in the male genital tract (ie, an excessive ROS production overwhelming all antioxidant strategies) impairs the structure and several functions of sperm cells. This phenomenon may arise in the female genital tract, but has never been investigated. The present paper is a review of the literature on these subjects and also reports our results concerning the changes in semen lipid content during cervical mucus migration and the effect of cervical mucus polymorphonuclear (PMN) cells on sperm characteristics. We showed that the sperm levels of vitamin E, cholesterol, phospholipids, sphingomyelin and plasmalogen assessed by HPLC decreased after migration through cervical mucus. These modifications were observed in parallel with lipid enrichment of the cervical mucus, suggesting an efflux of cholesterol and lipids from sperm cells. The spermatozoa recovered postmigration in the cervical mucus were characterised by low levels of the various lipid classes. Spermatozoa that migrated in cervical mucus samples with a considerable quantity of polymorphonuclear leukocytes (PMN) also showed significantly increased levels of sphingomyelin, diacyl phospholipids and plasmalogens in comparison to spermatozoa that migrated in cervical mucus devoid of PMN. Finally, we also found that PMA-induced ROS production was significantly increased for spermatozoa treated with cervical mucus containing PMN.     

Repeatability of the duration of oestrus and breed differences in the relationship between druation of oestrus and ovulation rate of sheep.

The duration of oestrus and the time interval from removal of progestagen-impregnated pessaries to the onset and end of oestrus were examined in Texel, Finnish Landrace, Galway and Fingalway (Finnish Landrace X Galway) ewes. The differences among the breeds in the relationship between these variables and ovulation rate at the controlled oestrus were also investigated. Breed differences were significant for all traits except the interval from pessary withdrawal to the onset of oestrus. The relationship between ovulation rate and both the interval from pessary withdrawal to the onset of oestrus and the duration of oestrus differed significantly among the breeds. The repeatability of the duration of oestrus was significant for Texel and Rambouillet ewes (mean = 0.5) and for pooled data from ewe lambs of various breeds. It was concluded that, in view of the breed differences in the relationship between ovulation rate and duration of oestrus and other traits, generalizations should not be made from among-breed to within-breed relationships. The high repeatability for the duration of oestrus may mean substantial heritabilities for the physiological determinants of oestrus duration.