Accuracy of evaluation of ovarian structures by transrectal ultrasonography in ewes

The objective of this study was to assess the accuracy of estimating the size and number of corpora lutea (CL) and ovarian follicles by ultrasonography (US) of the ewe in standing position. US observations were compared with those made at subsequent postmortem examination of the ovaries. Corriedale ewes (n=50) of unknown reproductive history and at random stages of the oestrus cycle were used for the study. Transrectal US was performed using a 7.5 MHz transducer with the ewe in standing position. The ewes were slaughtered 12 h after the US examination and the ovaries collected, dissected and the number and size of the CL and follicles evaluated. CL were classified as functional or non-functional on the basis of their colour and follicles were classified by size (2, 3, 4 and > or =5 mm). Accuracy of US was assessed by calculating its positive predictive value and sensitivity. The data were evaluated by Pearson correlation and linear regression analysis. The predictive value and sensitivity of US was 100% for the presence and 96% for the absence of CL. In four out of five ewes with double ovulations, the presence of both CL was correctly diagnosed by US. For functional CL, the sensitivity of US was 100%. The regression coefficient for CL diameter was significant (P< or =0.001; r2=0.4; n=35). The size of 85 of 117 follicles was accurately determined. The correlation between numbers of follicles counted by US and postmortem increased with size of follicle from r=0.44 (P< or =0.01) for 2 mm to r=0.85 (P< or =0.001) for >or =5 mm diameter follicles. The regression between the diameter of follicles determined by the two methods was significant (P< or =0.001; r2=0.8; n=117). The predictive value of US for the number of follicles was high (98-100%) for all follicle sizes except for 3 mm diameter follicles (predictive value 71%). Similarly, the sensitivity was high for all sizes of follicles (90-95%) except for those of 2 mm diameter (62%). It was concluded that ultrasound scanning provides a highly accurate method for determining the number of CL and follicles > or =4 mm diameter but that its predictive value and sensitivity are lower for smaller diameter follicles. The regression equation for diameter measured by US on that evaluated postmortem explained more of the variation for follicles than for CL.     

Determination of early pregnancy in ewes utilizing transrectal ultrasonography

Transrectal ultrasonography was used in ewes to determine the earliest day at which pregnancy could be detected, the number of embryos present, and the pattern of growth of the embryos. Twenty-one ewes were placed with 2 fertile rams and 20 ewes with 2 vasectomized rams. All ewes were treated to synchronize estrus and were observed for estrus twice daily. The 36 ewes that showed synchronized estrus were separated from the rams following mating. Transrectal ultrasonography was performed daily from estrus (Day 0) to Day 25 for all ewes and on Days 30, 35 and 40 post breeding for the 20 ewes mated to fertile rams. A 7.5 MHz transducer (human prostate, linear array) was utilized, with the ewes in dorsal recumbency in a tilting squeeze chute. Extraembryonic fluid and membranes were observed in the uterine horns ipsilateral to corpora lutea by Day 15 post breeding in all 17 ewes subsequently diagnosed as pregnant. Rhythmic pulsations (heartbeat) within the embryonic vesicles were first detected on Day 18 or 19. At least 1 embryo was detected by Day 20 in all the pregnant ewes, but not all the embryos were counted accurately until Day 25 (main effect of day; P < 0.05). Two ewes each had an embryo which died (absence of previously observed heartbeat) by Day 25 or Day 40, respectively, but each maintained the remaining embryos to term. The pattern of embryonic growth, as determined by crown-rump lengths on Days 20, 25, 30, 35 and 40, did not differ with the number of embryos carried (n = 1 to 4). In conclusion, transrectal ultrasonography was found to provide a rapid, accurate means for the early detection of pregnancy in ewes.     

Ultrasonographic study of ovarian function during early pregnancy and after parturition in the ewe

Transrectal ultrasonography of ovaries was performed in 11 ewes from Days 10 to 26 and on Day 30 of pregnancy to record the number, size, and position of ovarian follicles > or = 3 mm in diameter and corpora lutea (CL). Transrectal and/or transabdominal ultrasonography of the uterus was performed on Days 10 to 26, 30, 35, 40, 45 and 50 of gestation to ascertain the number and position of the conceptuses. In a second experiment, ultrasonography was conducted in 15 ewes on Days 10, 25, 30, 45 and 50 of pregnancy and from Days 13 to 29 after parturition. Ovarian data were classified into ovaries without CL (Group 1), ovaries with the CL in 1 ovary (Group 2), and ovaries with the CL in both ovaries during pregnancy (Group 3). In early pregnant ewes, the total number of follicles and the diameter of all follicles > or = 3 mm were smaller (P < 0.05) in the CL-bearing ovaries (both Group 2, n = 7 and Group 3, n = 8) than in the non-CL-bearing ovaries (Group 1, n = 7), while the largest follicle diameter was significantly smaller in Group 3 than in Group 1 or 2 ovaries. The number of 3-mm follicles in Group 2 ovaries was lower (P < 0.05) than in Group 1 or 3 ovaries, but the mean number of follicles > or = 5 mm in diameter was significantly lower in Group 3 than in Group 1. The total luteal volume per ovary was higher (P < 0.001) in Group 2 than in Group 3 ovaries of early pregnant ewes. The total follicle diameter and the number of follicles growing from 3 to > or = 5 mm in diameter was lower (P < 0.05) for Group 2 ovaries of ewes that carried twins (n = 3) compared with Group 2 ovaries from ewes with singletons (n = 4). There were no differences in follicular dynamics between Group 3 ovaries and the ovaries of Group 2 in ewes that carried twins. No follicles > 3 mm were seen in the ovaries of postpartum ewes that contained CL during gestation, until Days 21 and 25 postpartum for Groups 2 (n = 10) and 3 (n = 8), respectively, and follicles reaching > or = 5 mm in diameter were detected in only 2 ovaries (Group 2), on Days 27 and 28 postpartum, respectively. We conclude that during early pregnancy in ewes there is a suppression of antral follicle growth which appears to be exerted primarily by the developing conceptus but remains confined to CL-bearing ovaries. Residual local inhibition of follicular development extends into the postpartum period.     

Ultrasonographic study of the effects of the corpus luteum on antral follicular development in unilaterally ovulating western white-faced ewes

The objective of this study was to examine the local effects of the corpus luteum (CL) on ovarian antral follicle development by looking at follicle populations and dynamics in ovaries with or without CL, in unilaterally ovulating ewes, using a retrospective analysis of daily ultrasonographic records. The present report summarises the data from the first luteal phase of the breeding season (August-October; n = 4), a luteal phase in the mid-breeding season (November-December; n = 5), the last luteal phase of the breeding season (January-March; n = 5), and the luteal phase after GnRH-induced ovulations in mid-anoestrus (May-June; n = 4) of western white-faced ewes. Mean daily numbers of 3mm follicles that did not grow any larger were significantly reduced in the CL-containing ovaries of ewes at all periods of study except for the transition to anoestrus. With all scanning periods combined, daily numbers of 3mm follicles not growing further increased (P<0.05) between day 6 and 15 after ovulation in the CL-containing ovaries. Based on mean data for the whole periods of observation, the non-CL-bearing ovaries of ewes in the transition to anoestrus had fewer (P<0.05) follicles growing from 3 to > or =5mm in size before regression compared with the mid-breeding season and mid-anoestrus. The lifespan of follicles reaching > or =5mm in diameter was shorter (P < 0.05) in the CL- compared with non-CL-containing ovaries of anoestrous ewes induced to ovulate with GnRH ((6.5+/- 1.3) and (9.0+/- 1.0) days, respectively). Circulating concentrations of progesterone were lower during both transitional periods (into and out of anoestrus) and mid-anoestrus than during the mid-breeding season (P < 0.001), and were less during anoestrus than during both transitional periods (P < 0.05). It was concluded that CL/luteal structures locally suppressed the growth of ovarian antral follicles to the 3mm size-range except during the transition to anoestrus, but that there was no inhibitory effect of the CL on the growth of ovarian follicles to larger diameters. The presence of CL/luteal structures did not affect the length of the lifespan of follicles reaching > or =5mm in diameter nor the number of ovulations per ovary in cyclic ewes, but shortened large follicle lifespan in anoestrous ewes. Variations in peripheral concentrations of progesterone across the breeding season and between the breeding season and anoestrus did not alter the lifespan of large antral follicles. In the transition to anoestrus and during mid-anoestrus, the presence of the CL in an ovary appeared to maintain follicle development to ovulatory sizes and to increase the rate of turnover of large antral follicles, respectively.     

Ovarian function in ewes during the transition from breeding season to anoestrus

Transrectal ovarian ultrasonography was conducted in six Western white-faced ewes for 35 days from the last oestrus of the breeding season, to record the number and size of all ovarian follicles > or = 3 mm in diameter and luteal structures. Blood samples were collected once a day for estimation of serum concentrations of follicle-stimulating hormone (FSH), oestradiol and progesterone. Each ewe had five follicular waves (follicles growing from 3 to > or = 5 mm in diameter) over the scanning period. The duration of the growth phase of the largest ovarian follicles did not differ (P > 0.05) between waves, but follicular static and regressing phases decreased significantly (P < 0.05) after the decline in serum progesterone concentrations at the end of the last luteal phase of the breeding season. The intervals between the five follicular waves were: 9.2+/-0.4, 5.2+/-0.7, 8.3+/-0.8 and 5.8+/-0.7 days; the two shorter intervals differed (P < 0.05) from the two longer intervals. Using the cycle-detection program, rhythmic increases in serum FSH concentrations were detected in all ewes; the amplitude, duration and periodicity of FSH fluctuations did not vary (P > 0.05) throughout the period of study. The number of identified FSH peaks (7.8+/-0.5 peaks per ewe, per scanning period) was greater (P < 0.05) than the number of emerging follicular waves. Serum concentrations of oestradiol remained low (< or = 1 pg/ml) on most days, in five out of the six ewes studied, and sporadic elevations in oestradiol secretion above the non-detectable level were not associated with the emergence of follicular waves. The ovulation rate was lower than that seen during the middle portion of the breeding season (November-December) in white-faced ewes but the transitional ewes had larger corpora lutea (CL). Maximal serum concentrations of progesterone appeared to be lower and the plateau phase of progesterone secretion appeared to be shorter during the last luteal phase of the ovulatory season in comparison to the mid-breeding season of Western white-faced ewes. During the transition into anoestrus in ewes, the endogenous rhythm of FSH release is remarkably robust but the pattern of emergence of sequential follicular waves is dissociated from FSH and oestradiol secretion. Luteal progesterone secretion is suppressed because of fewer ovulations and diminished total luteal volume, but it may also result from diminished gonadotropic support. These season-related alterations in the normal pattern of ovine ovarian cycles appear to be due to reduction in ovarian responsiveness to gonadotropins and/or attenuation in secretion of luteinizing hormone (LH) occurring at the onset of the anovulatory season in ewes.     

Ovarian capillary blood flow in seasonally anoestrous ewes induced to ovulate by treatment with GnRH

The microsphere technique was used to obtain estimates of ovarian capillary blood flow near ovulation, in 8 seasonally anoestrous ewes, which were induced to ovulate by GnRH therapy. Plasma progesterone concentrations were monitored in jugular blood sampled between Days 4 and 7 after the onset of the preovulatory LH surge. The ewes were then slaughtered. Three of the ewes were treated with a single injection of 20 mg progesterone before GnRH therapy. In these ewes and 1 other, plasma progesterone values increased after ovulation and reached 1.0 ng/ml on Day 7 following the preovulatory LH surge (normal, functional CL), whilst in the other 4 ewes progesterone concentrations increased initially then declined to 0.5 ng/ml by Day 7 (abnormal CL). In the ewes exhibiting normal luteal function, the mean ovarian capillary blood flow was significantly greater (P less than 0.01) than that for ewes having abnormal luteal function. Irrespective of the type of CL produced, capillary blood flow was significantly greater (P less than 0.05) in ovulatory ovaries than in non-ovulatory ovaries. These findings indicate that the rate of capillary blood flow in ovaries near ovulation may be a critical factor in normal development and maturation of preovulatory follicles and function of subsequently formed CL.     

Response of anestrous ewes to the ram effect after follicular wave synchronization with a single dose of estradiol-17beta

Anestrous ewes respond to the introduction of rams with either an ovulation within 2-3 days that may be followed by luteal phases of normal or short length, with delayed ovulations (5-6 days later), or with the luteinization of follicles. The aim of this work was to study the relationship between the growth status of the largest follicle present when rams are introduced and the type of ovarian response in non-treated ewes and in ewes treated with estradiol-17beta before ram introduction. Thirteen anestrous Corriedale ewes were divided into 2 groups: E2 (n = 7) and C (n = 6). The E2 ewes received a single dose of 50 microg estradiol-17beta 5 days before the introduction of the rams to synchronize the onset of their follicle waves, while C ewes remained untreated. When the rams were introduced, all E2 ewes had the largest follicle in a growing stage in contrast with the C ewes (3 out of 6; P < 0.05). Five C and 4 E2 ewes ovulated after the introduction of the rams (Day 3.4 +/- 0.4 for C vs. 4.8 +/- 0.3 for E2 ewes, respectively, P < 0.05). Only one ewe from each group developed a normal luteal phase: 4 C and 3 E2 ewes had short luteal phases. One C ewe and 2 E2 ewes had short luteal phases originating from follicles that did not ovulate. After the first luteal phase, all ewes returned to anesirus without a second ovulation or luteal phase. The remaining E2 ewe did not ovulate or show any changes in progesterone serum concentrations. We conclude that the growth status of the largest follicle alone does not determine the ovarian responding pattern of anestrous ewes to the ram effect.     

Relationship of oestrus synchronization method, circulating hormones, luteinizing hormone and prostaglandin F-2 alpha receptors and luteal progesterone concentration to premature luteal regression in superovulated sheep

Ewes were treated with exogenous follicle-stimulating hormone (FSH) and oestrus was synchronized using either a dual prostaglandin F-2 alpha (PGF-2 alpha) injection regimen or pessaries impregnated with medroxy progesterone acetate (MAP). Natural cycling ewes served as controls. After oestrus or AI (Day 0), corpora lutea (CL) were enucleated surgically from the left and right ovaries on Days 3 and 6, respectively. The incidence of premature luteolysis was related (P less than 0.05) to PGF-2 alpha treatment and occurred in 7 of 8 ewes compared with 0 of 4 controls and 1 of 8 MAP-exposed females. Sheep with regressing CL had lower circulating and intraluteal progesterone concentrations and fewer total and small dissociated luteal cells on Day 3 than gonadotrophin-treated counterparts with normal CL. Progesterone concentration in the serum and luteal tissue was higher (P less than 0.05) in gonadotrophin-treated ewes with normal CL than in the controls; but luteinizing hormone (LH) receptors/cell were not different on Days 3 and 6. There were no apparent differences in the temporal patterns of circulating oestradiol-17 beta, FSH and LH. High progesterone in gonadotrophin-treated ewes with normal CL coincided with an increase in total luteal mass and numbers of cells, which were primarily reflected in more small luteal cells than in control ewes. Gonadotrophin-treated ewes with regressing CL on Day 3 tended (P less than 0.10) to have fewer small luteal cells and fewer (P less than 0.05) low-affinity PGF-2 alpha binding sites than sheep with normal CL. By Day 6, luteal integrity and cell viability was absent in ewes with prematurely regressed CL. These data demonstrate that (i) the incidence of premature luteal regression is highly correlated with the use of PGF-2 alpha; (ii) this abnormal luteal tissue is functionally competent for 2-3 days after ovulation, but deteriorates rapidly thereafter and (iii) luteal-dysfunctioning ewes experience a reduction in numbers of small luteal cells without a significant change in luteal mass by Day 3 and, overall, have fewer low-affinity PGF-2 alpha binding sites.     

Real-time ultrasonography for determining pregnancy status and viable fetal numbers in ewes

The reproductive tracts of 26 estrus synchronized, bred ewes were scanned with a portable 5.0 MHz real-time ultrasound unit within 1 to 6 d postbreeding. Intrarectal scanning was performed on alternate days until Days 28 to 30 and twice weekly until Days 50 of gestation. Transabdominal uterine scans were conducted twice weekly from Days 25 to 65 and continued weekly until parturition. A total of 24 ewes (92%) became pregnant. A nonpregnant ewe was recognized 100% of the time by both methods of ultrasonic screening. Correct identification of a gravid ewe as pregnant was 100% from Days 51 to 150 of gestation using transabdominal real-time ultrasonography. There was a significant association (P < 0.005) between the number of lambs born and the number of fetuses observed using transabdominal real-time ultrasonography after Day 25 of gestation. Accurate differentiation of fetal numbers by transabdominal scanning was 100.0% for ewes carrying one lamb and 97.3% for ewes carrying two lambs at Days 51 to 75 of gestation. Fetal attrition was documented in one ewe at Day 49 of gestation. Hydrops allantois was diagnosed in another ewe at 110 d of gestation. A total of 37 lambs were born to 23 ewes in the project flock. No congenital abnormalities were noted in any of the lambs. Transabdominal real-time ultrasonography is a safe, rapid, accurate and practical method for assessing pregnancy status, fetal number and fetal viability in sheep.     

Effect of diet and GnRH administration on post-partum ovarian cyclicity in autumn-lambing ewes

Using autumn-lambing ewes, this study investigated (i) the effects of diet on gonadotrophin secretion and responsiveness of the hypothalamic-pituitary-ovarian axis to exogenous GnRH during the early post-partum period; and (ii) whether ovulation prior to completion of uterine involution results in an increased incidence of aberrant ovarian cycles. Thirty-two ewes rearing 1.9+/-0.12 lambs were equally allocated to two dietary treatments at lambing (22 October +/-0.2 day). Diets comprised ad libitum hay and 1.5 kg per ewe per day of one of two concentrates (11.5 MJ ME, 195 g CP per kg) containing 300 g kg(-1) cracked maize grain (M) or 300 g kg(-1) sugar beet pellets (S). Half of the ewes on each diet (G) received 25 i.v. injections of 250 ng GnRH in 2 ml 0.9% saline at 2 h intervals from days 12-14 post-partum while remaining ewes (N) were monitored for the resumption of spontaneous ovarian cyclicity. Blood samples were obtained from all ewes throughout the study (lambing to 18 December) for measurement of circulating hormone concentrations and the uteri and ovaries of all ewes were examined via laparoscopy on day 21 post-partum. There were no effects of dietary treatment on ewe daily live weight loss, lamb daily live weight gain or the immediate post-partum increase in circulating FSH concentrations. Diet did not affect insulin concentrations or LH pulse frequency on day 12 post-partum but LH pulse amplitude was lower in ewes fed concentrate M compared to concentrate S (1.4+/-0.10 versus 1.7+/-0.12 ng ml(-1), respectively, P<0.05) and this was associated with an increased interval to the resumption of spontaneous ovarian cycles (35+/-3.1 versus 26+/-2.1 day, respectively, P<0.05). Administration of exogenous GnRH increased (P<0.05) the proportion of ewes on both diets that ovulated within 20 days of parturition and advanced the onset of ovarian cyclicity in ewes fed concentrate M by 9.5 days (significance of interaction, P<0.05). Four ewes, all of which ovulated before day 22 post-partum, had extended luteal activity while in remaining ewes, duration of the first luteal phase was inversely related to the time of first ovulation (r(2)=0.16, P<0.05). Results demonstrate that (i) the onset of ovarian cyclicity is influenced by diet and can be advanced by administration of exogenous GnRH; and (ii) ovulation during the early post-partum period is associated with an increased incidence of extended luteal activity.     

Ovarian function in ewes at the onset of the breeding season

Transrectal ultrasonography of ovaries was performed each day, during the expected transition from anoestrus to the breeding season (mid-August to early October), in six Western white-faced cross-bred ewes, to record ovarian antral follicles > or = 3 mm in size and luteal structures. Jugular blood samples were collected daily for radioimmunoassay (RIA) of follicle-stimulating hormone (FSH), oestradiol and progesterone. The first ovulation of the breeding season was followed by the full-length oestrous cycle in all ewes studied. Prior to the ovulation, all ewes exhibited a distinct increase in circulating concentrations of progesterone, yet no corpora lutea (CL) were detected and luteinized unovulated follicles were detected in only three ewes. Secretion of FSH was not affected by the cessation of anoestrus and peaks of episodic FSH fluctuations were associated with the emergence of ovarian follicular waves (follicles growing from 3 to > or = 5 mm). During the 17 days prior to the first ovulation of the breeding season, there were no apparent changes in the pattern of emergence of follicular waves. Mean daily numbers of small antral follicles (not growing beyond 3 mm in diameter) declined (P < 0.05) after the first ovulation. The ovulation rate, maximal total and mean luteal volumes and maximal serum progesterone concentrations, but not mean diameters of ovulatory follicles, were ostensibly lower during the first oestrous cycle of the breeding season compared with the mid-breeding season of Western white-faced ewes. Oestradiol secretion by ovarian follicles appeared to be fully restored, compared with anoestrous ewes, but it was not synchronized with the growth of the largest antral follicles of waves until after the beginning of the first oestrous cycle. An increase in progesterone secretion preceding the first ovulation of the breeding season does not result, as previously suggested, from the ovulation of immature ovarian follicles and short-lived CL, but progesterone may be produced by luteinized unovulated follicles and/or interstitial tissue of unknown origin. This increase in serum concentrations of progesterone does not alter the pattern of follicular wave development, hence it seems to be important mainly for inducing oestrous behaviour, synchronizing it with the preovulatory surge of luteinizing hormone (LH), and preventing premature luteolysis during the ensuing luteal phase. Progesterone may also enhance ovarian follicular responsiveness to circulating gonadotropins through a local mechanism.     

The introduction of rams induces an increase in pulsatile LH secretion in cyclic ewes during the breeding season

Application of the ram effect during the breeding season has been previously disregarded because the ewe reproductive axis is powerfully inhibited by luteal phase progesterone concentrations. However, anovulatory ewes treated with exogenous progestagens respond to ram introduction with an increase in LH concentrations. We therefore tested whether cyclic ewes would respond to ram introduction with an increase in pulsatile LH secretion at all stages of the estrous cycle. We did two experiments using genotypes native to temperate or Mediterranean regions. In Experiment 1 (UK), 12 randomly cycling, North of England Mule ewes were introduced to rams midway through a frequent blood-sampling regime. Ewes in the early (EL; n=3) [corrected] and late luteal (LL; n=6) phase responded to ram introduction with an increase in LH pulse frequency and mean and basal concentration [corrected] of LH (at least P<0.05). In Experiment 2 (Australia), the cycles of 32 Merino ewes were synchronised using intravaginal progestagen pessaries. Pessary insertion was staggered to produce eight ewes at each stage of the estrous cycle: follicular (F), early luteal (EL), mid-luteal (ML) and late luteal (LL). In all stages of the cycle, ewes responded to ram introduction with an increase in LH pulse frequency (P<0.01); EL, ML and LL ewes also had an increase in mean LH concentration (P<0.05). In conclusion, ram introduction to cyclic ewes stimulated an increase in pulsatile LH secretion, independent of ewe genotype or stage of the estrous cycle.     

The relationship between vaginal mucous impedance and serum concentrations of estradiol and progesterone throughout the sheep estrous cycle

The objective of this experiment was to assess the relationship between electrical resistance of the vaginal mucosa and serum concentrations of estradiol (E2) and progesterone (P4) during the estrous cycle in ewes. Vaginal impedance was recorded daily using a 2-electrode impedometer in 10 nonprolific Western white-faced and 7 prolific Finn ewes, during the mid-breeding season (October to December). Transrectal ultrasonography of ovaries was performed once a day to confirm ovulation and monitor follicle growth (follicles > or =3 mm in diameter) and development of corpora lutea (CL). Jugular blood samples were collected daily for radioimmunoassay (RIA) of estradiol and progesterone. In all ewes, a decline in vaginal impedance (to <40 ohms) was closely associated with the onset of behavioral estrus. In both breeds of sheep, there was no significant correlation between daily serum concentrations of estradiol and vaginal impedance throughout the estrous cycle. Daily serum concentrations of progesterone and the E2:P4 ratio were correlated with vaginal impedance during the period of luteolysis and follicular phase in both breeds (Western white-faced ewes: r = 0.62, P = 0.0002 and r = -0.56, P = 0.0002; Finn ewes: r = 0.61, P = 0.001 and r = -0.45, P = 0.03, respectively) and early in the cycle (Days 0 to 2, Day 0 = day of ovulation) in white-faced ewes (r = 0.61, P = 0.0003 and r = -0.36, P = 0.052, respectively) but not during the remaining portion of the luteal phase in either breed. In conclusion, vaginal mucous impedance appears to be primarily controlled by progesterone, but it also changes in response to shifts in the E2:P4 ratio when progesterone concentrations are low. Impedometric characteristics of the vaginal mucosa in cyclic ewes are an indicator of serum concentrations of progesterone and E2:P4 ratios during the terminal stage of the estrous cycle.     

Evaluation of gross anatomical features of cervix of tropical sheep using cervical silicone moulds

The lambing rate obtained following cervical artificial insemination (AI) with frozen semen in sheep is low mainly due to the inability of frozen-thawed sperm to traverse the tortuous nature of the cervical canal. Although acceptable fertility has been attained by circumventing the cervical barrier through laparoscope aided intrauterine AI, the emphasis is currently given on the development of alternate non-invasive transcervical AI procedures. The complex anatomy of the cervix does not facilitate easy transcervical passage for an insemination catheter. The aim of the present study was: (i) to examine the gross anatomy of the cervix in slaughtered ewe lambs and adult ewes of the native Malpura and Kheri breeds raised under semi-arid tropical environment; and (ii) to cast silicone moulds of the reproductive tracts for measuring the dimensions of the cervix. Eighty reproductive tracts were excised immediately from carcass of Malpura and Kheri ewes and the external os of each one was classified depending on their appearance as duckbill, spiral, rosette or flap. The cervical canal of each tract was filled with a silicone sealant for casting the mould. Fifty complete silicone moulds were obtained representing 25 from ewe lambs and 25 from adult ewes. The mean lengths of the cervical mould of ewe lambs and adult ewes were 3.8+/-0.12 and 5.3+/-0.15 cm, respectively. The average number of funnel shaped folds in the cervical mould of ewe lambs and adult ewes were 3.2+/-0.19 and 3.4+/-0.22. However, the second and third-folds from the os were observed to be accentric in both ewe lambs and adult ewes. The information generated in this study would be useful for increasing the success rate of penetration in ewes exhibiting estrus in order to improve the lambing rate of tropical ewes following transcervical AI.     

Corpus luteum function in ewes stimulated by rams

In late February Dorset rams were introduced (day = 0) to 40 mature Romney ewes that were observed by laparoscopy to be anovular. The ovaries of 20 of these ewes were examined by laparoscopy every second day while the remaining 20 ewes served as unoperated controls. Jugular blood samples were taken daily and plasma progesterone concentrations assayed to provide information on the functional status of any corpora lutea (CL) arising from ovulations stimulated by introduction of the rams. Eighty-five percent (-17/20) of the ewes that were repeatedly laparoscoped had ovulated within 4 days of ram introduction and premature regression of the CL had occurred between days 4 and 8 in 8 ewes and days 6 to 10 in 2 ewes. A second ovulation was observed after or during the premature regression of the first CL and this subsequent CL was maintained for the normal duration. The prematurely regressing CL produced a small peak in progesterone concentration on days 4 to 5 but the concentrations declined on days 6 to 7. In the unoperated ewes 50% (-10/20) appeared, from the progesterone profiles, to have ovulated by day 4 and half of these appeared to have premature CL regression. The interval from introduction of the ram to first oestrus was 23 days in ewes with premature regression of the CL and 19 days in ewes ovulating within 4 days but having no premature regression. From the results it was concluded that the premature regression of the CL is the cause of the delayed interval from ram introduction to first oestrus in Romney ewes and is a major factor contributing to the two peaks of oestrous activity observed after ram introduction.     

Ultrasound-guided transvaginal oocyte collection in prepubertal calves

The present study was designed to develop a technique for oocyte collection using an ultrasound-guided transvaginal approach in prepubertal calves too small to accommodate manual transrectal manipulation. A commercially available, 5 MHz, convex-array ultrasound transducer designed for intravaginal use in women was custom-modified for use in calves. In Experiment 1, calves 10 to 16 wk old (n = 10) were restrained in the standing position in an adjustable squeeze chute with regional anesthesia. In Experiment 2, the follicle aspiration procedure was performed in 6 wk-old calves (n = 20) in dorsal recumbency after tranquilization and caudal epidural anesthesia. Ovarian superstimulation was induced in half of the calves using 750 IU eCG (Experiment 1) or 200 mg Folltropin (Experiment 2). Consistent visualization of both ovaries using the transvaginal approach was accomplished after considerable practice. Two methods of ovarian immobilization were attempted, but both interfered with the ultrasound image and were consequently abandoned. The inability to immobilize the ovary resulted in attempts to spear the intended follicle "free-hand." The contents of follicles >or= 6 mm in diameter were aspirated, filtered, and oocytes were located using a stereomicroscope. Although ovarian superstimulation did not resolve the problem of ovary movement, the number of follicles of adequate size to aspirate was dramatically increased. Extremely sharp needles were found to be very important in the free-hand technique. A total of 232 follicles was aspirated, and 100 oocytes were collected (43%). In summary, a transvaginal ultrasound-guided technique for oocyte collection was developed for young calves in a standing position (10 to 16 wk of age) and dorsal recumbency (6 wk of age). Results demonstrate the potential utility of this approach for deriving oocytes from young calves.     

Comparisons of the ovulatory and steroidogenic activities of the left and right ovaries of the ewe

The ovary in which a CL was observed by laparoscopy in Finnish Landrace, Tasmanian Merino and Scottish Blackface ewes had no apparent effect on the location of the CL during the succeeding oestrous cycles, on the duration of the associated oestrous cycles, or on the peripheral progesterone concentrations on Days 7 and 11. Although 53% of CL were present in the right ovaries, the difference between the two sides was not significant.     

Determination of gestational age in sheep and goats using transrectal ultrasonographic measurement of placentomes

Three experiments were conducted to determine gestational age in the ewe and doe by measuring placentomes with a B-mode ultrasonograph and a 5 MHz transducer. Transrectal measurements were obtained by placing the female over a bale of hay. In Experiment 1, ewes (n = 12) and does (n = 15) were examined by transrectal ultrasonography every week from breeding to parturition to determine the growth pattern of placentomes during pregnancy. In Experiment 2, placentomes from 132 ewes and 169 does were measured between 30 and 90 d of gestation. A linear regression relationship between fetal age in days and placentome size in mm was calculated and adjusted for does (gestational age = 28.74 + 1.80PL + e, r(2) = 70.34) and for ewes (age = 47.98 + 0.62PL + e, r(2) = 15.59). In Experiment 3, the placentomes of 63 does were measured to validate this relationship by using linear regression. Gestational age was determined correctly in 66% of the does, with a range of +/- 7 d and in 96% with a margin of +/- 14 d. In conclusion, transrectal ultrasonography allowed for the measurement of placentome size, which increased rapidly during the first 70 to 90 d of gestation in ewes and does. In ewes, however, there was a poor correlation of placentome size with gestational age, while in goats, measurement of placentomes could be used along with pregnancy diagnosis by transrectal ultrasonography as an indication of gestation age.     

Ultrasonographic and laparoscopic evaluation of the reproductive tract of the captive female African lion (Panthera leo)

The use of transabdominal ultrasonography to assess the oestrous cycle has not been previously described in the African lion (Panthera leo). Twelve sexually mature lionesses and five female cubs had their reproductive organs assessed by transabdominal ultrasound. Ovarian findings were compared to laparoscopic findings while performing laparoscopic ovariectomy or salpingectomy. Vaginal cytology was performed and serum progesterone levels were determined. By combining all data the oestrous cycle stage of each lion was determined. One lion was far pregnant and was not operated on. In adults a uterine body could be seen ultrasonographically in 67% of lions while mural structures could be distinguished in 44% of lions. Five uterine horns could be seen in 3 lions. In 12 adults 10 ovaries were found of which eight had discernable follicles or luteal structures. During laparoscopy 12 active ovaries were seen with luteal structures seen in 11 ovaries and follicles in 2 ovaries. Using laparoscopy as the gold standard, ultrasonography had a sensitivity of 66% and specificity of 83% to detect ovarian reproductive activity. Two uterine cysts and a cluster of periovarian cysts were seen in three different lions. Three lions were pregnant, two were in oestrus, three in a luteal phase (dioestrus), and four were in anoestrus. Transabdominal ultrasound in combination with serum progesterone levels and vaginal cytology can be used to assess ovarian cyclical activity with reasonable accuracy in captive bred lions.