Oestrus cycle characteristics and prediction of ovulation in Catalonian jennies

The Catalonian donkey breed is in danger of extinction, and much needs to be learned about the reproductive features of its females if breeding and conservation programmes are to be successful. This study reports the oestrous behaviour, oestrus cycle characteristics and dynamic ovarian events witnessed during 50 oestrous cycles (involving 106 ovulations) in 10 Catalonian jennies between March 2002 and January 2005. These jennies were teased, palpated transrectally and examined by ultrasound using a 5 MHz linear transducer—daily during oestrus and every other day during dioestrus. Predictors of ovulation were sought among the variables recorded.The most evident signs of oestrus were mouth clapping (the frequent vertical opening and closing of the mouth with ears depressed against the extended neck) and occasional urinating and winking of the vulval lips (homotypical behaviour). Interactions between jennies in oestrus were also recorded, including mounting, herding/chasing, the Flehmen response, and vocalization (heterotypical behaviour).Nine jennies ovulated regularly throughout the year; one had two anovulatory periods (54 and 35 days). The length of the oestrus cycle was 24.90 ± 0.26 days, with oestrus itself lasting 5.64 ± 0.20 days (mean ± S.E.M.) and dioestrus 19.83 ± 0.36 days. The incidence of single, double and triple ovulations was 55.66% (n = 59), 42.45% (n = 45) and 1.89% (n = 2), respectively. No significant difference was seen in the number of ovulations involving the left and right ovaries (52.63% [n = 70] compared to 47.37% [n = 63] respectively; P > 0.05). The mean interval between double ovulation was 1.44 ± 3.98 days. The mean diameter of the preovulatory follicle at day −1 was 44.9 ± 0.5 mm; the mean growth rate over the 5 days before ovulation was 3.7 mm/day.Data on preovulatory changes in oestrous behaviour, follicle size, follicle texture, the echographic appearance of the follicle and uterus, and uterine tone were subjected to stepwise logistic regression analysis to detect predictors of ovulation. The logit function showed the best predictors to be follicle size, follicular texture and oestrous behaviour. Certain combinations of these three variables allow the prediction of ovulation within 24 h with a probability of >75%.     

Ovarian follicular development in Holstein cows following synchronisation of oestrus with oestradiol benzoate and an intravaginal progesterone releasing insert for 5-9 days and duration of the oestrous cycle and concentrations of progesterone following ovulation

The aim of this study was to determine if the duration of treatment with an intravaginal progesterone releasing insert (IVP(4)) after treatment with oestradiol benzoate (ODB) at the time of insertion and 24 h after removal would affect selected variables including: size of ovarian follicles at the time of removal of inserts, diameter of ovulatory follicles, plasma concentrations of progesterone following ovulation, and duration of the following oestrous cycle. Characteristics of oestrus at a synchronised and spontaneous oestrus were also monitored. Non-lactating Holstein cows were synchronised with an IVP(4) for 5 (n = 10), 7 (n = 10), 8 (n = 9) or 9 (n = 9) days together with injections of ODB at device insertion (2 mg) and 24 h after removal (1 mg). Ultrasonography showed no significant effect of treatment on the day of emergence of preovulatory follicles relative to the day of removal of inserts (overall mean = -4.22 +/- 0.58; P = 0.15) for cows that ovulated within 120 h insert removal (n = 36). Treatment with ODB and an IVP(4) for 5 days reduced the diameter of preovulatory follicles at the time of removal of inserts and for the following 2 days compared to cows treated for 7-9 days (mean difference 2.56 +/- 1.15 mm; P = 0.033) but did not reduce the diameter of the ovulatory follicle (P = 0.21). Day of emergence relative to removal of inserts was associated with the diameter of the ovulatory follicle (R2 = 0.69; P < 0.001). Concentrations of progesterone and the diameter of the corpus luteum following ovulation were not affected by treatment (P > 0.20), but were affected by the diameter of the ovulatory follicle (P < 0.01). Diameter of the ovulatory follicle did not affect interoestrous and interovulatory intervals (P > 0.40). We conclude that treatment with an IVP(4) for 5 compared to 7-9 days with ODB administered at device insertion, and 24 h after removal reduced the diameter of preovulatory follicles at the time of removal of the insert but did not reduce the diameter of the ovulatory follicle or concentrations of progesterone in plasma. Emergence of preovulatory follicles closer to the time of removal of inserts reduced the diameter of the ovulatory follicle when oestrus was induced with ODB. Ovulation of smaller follicles reduced concentrations of progesterone in plasma following ovulation but did not affect oestrous cycle duration.     

Ultrasonographic evaluation of uterine involution and postpartum follicular dynamics in French jennies (Equus asinus)

Uterine involution and follicular dynamics during postpartum period were studied ultrasonographically in French jennies. For the study of uterine involution in postpartum jennies (n = 6, Group S), sonographic measurements of different parts of the uterus and endometrium were made at three-day interval, starting from the day of foaling and continued up to 33 days postpartum. Uterine dimensions were also recorded in non-pregnant jennies (n = 3, Group C) throughout a cycle and compared with the dimensions of Group S jennies observed on the day of complete involution. Follicular dynamics of first and second postpartum ovulatory cycles were studied and compared with that of the single estrous cycle of Group C jennies. Jugular venous blood samples of Group S jennies were collected at weekly intervals for 49 days, commencing at the appearance of first preovulatory follicle, to support the sonographic findings. The average involution period was 22.5 +/- 1.7 days. However, it was significantly delayed (P < 0.05) in jennies which came into first postpartum ovulatory heat within Day 9 than those who came later (25.0 +/- 1.0 versus 20.0 +/- 1.0). The endometrial layer was not discernible beyond Day 15 postpartum and thus was found to be unreliable index of uterine involution. The follicular growth rate (mm per day) and diameter (mm) of preovulatory follicle in postpartum jennies were similar to that in normal cycling jennies (P > 0.05). The first and second ovulations occurred at 14.6 +/- 0.8 and 39.0 +/- 0.8 days postpartum in Group S jennies. All the corpora lutea, either echogenic or centrally non-echogenic were functionally similar and had similar life span (P > 0.05). In conclusion, the postpartum reproductive events related to uterine involution and ovarian cyclicity apparently resemble that of mares.     

Behavioral, follicular and gonadotropin changes during the estrous cycle in donkeys

Sexual behavior, follicular development and ovulation, and concentrations of circulating gonadotropins during the estrous cycle were studied during the summer in 7 jennies. Mean behavioral estrous length was 6.4 +/- 0.6 days (mean +/- SEM, n=19; 5.6 +/- 0.5 days preovulatory and 0.8 +/- 0.2 days post-ovulatory). Mean diestrous length was 19.3 +/- 0.6 days (n=14). Females in estrus typically showed posturing, mouth clapping, clitoral winking, urinating and tail raising. Mouth clapping began approximately one day sooner and lasted approximately one day longer than winking and tail raising, so that the total duration of clapping was significantly greater than for the other two signs. Follicular changes and concentrations of gonadotropins were determined for 14 estrous cycles (2 per jenny). The follicular end points [diameter of the largest follicle and number of large (>25 mm), medium (20-24 mm), and small follicles (<20 mm)] showed a significant day effect. The diameter of the largest follicle and the number of large follicles began to increase significantly 7 days prior to ovulation with a maximum value the day before ovulation. Medium follicles reached a maximum number 4 days prior to ovulation, and small follicles decreased significantly prior to ovulation. After ovulation, all follicular end points, except the number of small follicles, remained low for the next 12 days. Mean values of FSH were low during estrus and high during diestrus with 2 significant peaks, one 3 days and one 9 days after ovulation. In contrast, mean levels of LH were low during diestrus and high during estrus with a maximum value the day after ovulation. The LH profile showed a more prolonged gradual increase prior to ovulation, than that which has been reported for ponies and horses.     

Pattern of follicular growth and resumption of ovarian activity in post-partum beef suckler cows

The ovaries of 18 post-partum beef suckler cows were examined daily, using ultrasound, from Day 5 post partum until a normal oestrous cycle was completed. Periods of growth and regression of medium-sized (5-9 mm) follicles were identified before one medium follicle became dominant (single large follicle greater than or equal to 10 mm). The mean (+/- s.e.m.) number of days from parturition to detection of the first post-partum dominant follicle was 10.2 +/- 0.5. The first post-partum dominant follicle ovulated in 2/18 (11%) cows. The interval from calving to first ovulation (mean +/- s.e.m. = 35.9 +/- 3.3 days) was characterized by the growth and regression of a variable number (mean = 3.2 +/- 0.2; range 1-6) of dominant follicles. The maximum diameter of the dominant follicle increased as the cows approached first ovulation (P less than 0.05). Behavioural oestrus was not detected in 16/18 (89%) cows at first ovulation. Following first ovulation, the length of the subsequent cycle was short (mean = 9.7 +/- 0.5 days; range 8-15 days) in 14/18 (78%) cows and was characterized by the development and ovulation of a single dominant follicle. During oestrous cycles of normal length (mean = 20.6 +/- 0.5 days; range 18-23 days) one (N = 2), two (N = 7) or three (N = 8) dominant follicles were identified. The growth rate, maximum diameter or persistence of non-ovulatory dominant follicles before first ovulation or during oestrous cycles were not different (P greater than 0.05). These data show that, in beef suckler cows, follicular development and formation of a dominant follicle occur early after parturition and the incidence of ovulation of the first dominant follicle is low. The number of dominant follicles that develop before first ovulation is variable; first ovulation is rarely associated with oestrus and short cycles are common after first ovulation. It is concluded that prolonged anoestrus in post-partum beef suckler cows is due to lack of ovulation of a dominant follicle rather than delayed development of dominant follicles.     

Time of ovarian follicular recruitment in cyclic pony mares

An experiment was carried out on pony mares to establish the time of the oestrous cycle at which ovarian follicles are recruited for ovulation. In one group (n=7), the cycle was interrupted at the preovulatory stage by removing the preovulatory follicle; in another group (n=13) the cycle was interrupted at day 6 of the luteal phase by inducing luteolysis with a prostaglandin injection (PG). In a subgroup (n=7) of those given PG, the ovary not bearing the corpus luteum was removed at the time of injection. A further group (n=6) served as surgical controls. The interval to the next ovulation and blood concentrations of FSH were observed. Anaesthesia alone induced in preovulatory mares was followed by normal ovulation 2.5+/-1 days later. Removal of the preovulatory follicle delayed the next ovulation (14.6+/-2.1 days; P < 0.01). Following PG injection, the interval to ovulation was similar regardless of whether an ovary was removed (12.8+/-4.3 days) or not (10+/-4.1 days). This similarity occurred despite a large and prolonged rise in plasma FSH levels that occurred only in the hemiovariectomized group. In addition, the intervals found after PG injection did not differ from those found after ablation of the preovulatory follicle. These observations indicate that 1) in the presence of the early active corpus luteum or dominant follicle, follicles grow to a similar stage of development; 2) recruitment of the follicle due to ovulation occurs 12 to 14 days before ovulation; 3) limiting new follicular growth to one ovary does not affect the time course to ovulation; and 4) prolonged high FSH levels do not alter the time course or ovulation rate.     

Time of ovulation in nulliparous and multiparous goats

Fifteen nulliparous and nine multiparous Serrana goats were used, through two successive oestrous cycles, in order to characterize their ovulation time with regard to the number of ovulations after induced and natural oestrus during the breeding season. The onset of oestrus was detected by the amount of vasectomized bucks after oestrus synchronization with prostaglandin, given 10 days apart, and in the following two expected natural oestrus. The preovulatory LH peak was determined from blood samples collected 0, 4, 8, 12, 16, 20 and 24 h after onset of oestrus. A transrectal ovarian ultrasound scanning was performed 20, 24, 28, 32, 36, 40, 44 and 60 h after onset of oestrus, for the detection of ovulations by means of the disappearance of large follicles (>4 to 5 mm). Single ovulations were observed in 76% of oestrous periods in nulliparous goats and in 18% of nulliparous goats. The onset of oestrus to LH peak interval was lower in nulliparous (12.1 ± 0.9 h, n = 38) than in multiparous (15.6 ± 1.0 h, n = 22, P < 0.05) goats with no oestrus interaction effects (P > 0.05). The LH peak to first ovulation interval was higher after natural (18.9 ± 0.7 h, n = 36) than after induced (15.8 ± 1.2 h, n = 24, P < 0.05) oestrus. The onset of oestrus to total ovulation interval was influenced by parity (P < 0.01) and oestrus type (P < 0.05) with a length of 30.1 ± 1.1 h (n = 15) and 33.4 ± 1.5 h (n = 9) for induced oestrus of nulliparous and multiparous goats, respectively, and 32.5 ± 1.0 h (n = 23) and 36.5 ± 1.1 h (n = 13) for natural oestrus of nulliparous and multiparous goats, respectively. The onset of oestrus to first ovulation interval was not influenced by parity, but an interval of 8.0 ± 1.6 h was observed between the first and second ovulations in polyovulatory oestrus. Consequently, nulliparous goats that are predominantly monovular ovulate earlier than multiparous goats that are predominantly polyovulatory. In conclusion, significant differences occurred in the number and time of ovulations between nulliparous and multiparous goats. More research is necessary for a deeper understanding of the mechanisms regulating monovularory and polyovulatory oestrous cycles regarding the parity of goats.     

Ultrasonic evaluation of the preovulatory follicle in the mare

Ultrasonically visible characteristics of preovulatory follicles in mares which single ovulated were studied daily for 79 preovulatory periods in 40 mares. The preovulatory follicle became the largest follicle in the ovary from which ovulation later occurred six or more days before ovulation in 65 of 79 (82%) preovulatory periods; the mean was day -7 (range, day -14 to day -4). The increase in mean diameter of the preovulatory follicle was linear (R(2)=99.5%) over day -7 (29.4 +/- 0.8 mm) to day -1 (45.2 +/- 0.5 mm; growth rate, 2.7 mm/day). Follicles which double-ovulated were smaller (P<0.05) on day -1 (36 +/- 1.6 mm; n=12 follicles). Preovulatory follicles exhibited a pronounced change in shape from a spherical to a conical or pear-shaped structure in 84% of the preovulatory periods. Remaining follicles retained a spherical shape. Scores representing thickness of the follicular wall increased (P<0.05) as the interval to ovulation decreased. There was no significant difference among days in mean gray-scale value of the follicular wall or in echogenicity of the follicular fluid. Although diameter and shape of the follicle and thickness of the follicular wall changed during the preovulatory period, no reliable ultrasonically visible predictor of impending ovulation was found.     

Relationships between sexual behaviour, dominant follicle area, uterus ultrasonic image and pregnancy rate in mares of two breeds differing in reproductive efficiency

Weak or equivocal expression of oestrous behaviour, related to different level of mares' reactivity, may cause problems in oestrus detection and thus influence the reproductive efficiency. The aim of the study was to test whether a breed characterised by low pregnancy rate and high emotional reactivity (Thoroughbreds) differs in oestrous behaviour from a primitive breed with higher reproductive efficiency (Koniks). Additionally, the follicle size was examined to determine how it influences the intensity of oestrus and uterine ultrasonic image and whether the pregnancy rate differs in relation to the intensity of oestrous behaviour and the size of the dominant follicle(s) area on day preceding ovulation in both breeds. During four reproductive seasons the behaviour of 20 Konik polski (K) and 37 Thoroughbred (T) was observed during daily teasing. Simultaneously, the ultrasonic examinations of the reproductive system were carried out. The behaviour of mares was quantified by scoring on an 8-point scale (behavioural score, BS), according to increasing sexual receptivity. Cross-sectional follicular area (FA) was taken as a product of the two largest perpendicular follicular diameters and mean values for each breed were estimated on 1693 and 1982 mm2 for K and T mares, respectively (P < 0.05). Mares were classified according to the pooled area of dominant follicle (FA) during the preovulatory period: group A (FA < or = breed mean) and group B (FA > breed mean). Uterus image (UI) was scored (1-5) according to the increasing uterine echogenicity. The BS was higher (P < 0.01) in K mares (BS = 5.19) than in T mares (BS = 4.04). The BS was significantly related to increasing follicular area (FA). There was no breed difference in uterus image (UI) score. However, significant regression of UI on FA was found in K mares. The intensity of oestrus was positively related with UI (r = 0.29; P < 0.01) only in K mares, no such relationship was found in T mares. The pregnancy rate was significantly higher for K mares (88.5%) than for T mares (46.0%) and lower for T mares with less intense oestrous behaviour (29.4%) as compared to T mares with more intense oestrus (60.0%). No differences in pregnancy rate was found in mares belonging to A or B group of follicular area. The ascertained weaker oestrous behaviour in Thoroughbred was related to lower pregnancy rate. It is hypothesised that oestrus intensity may be the result of breed differences in the response of the neural system to follicular secretions, or may be an effect of higher incidence of multiple non-synchronic ovulation and/or higher sensitivity to stress in Thoroughbred mares.     

The sow endometrium at different stages of the oestrous cycle: studies on the distribution of CD2, CD4, CD8 and MHC class II expressing cells

The aim of this study was to investigate the distribution of CD2+, CD4+, CD8+ lymphocyte subpopulations and MHC class II expressing cells in the sow endometrium throughout the oestrous cycle. Fifteen crossbred multiparous sows (Swedish Landrace x Swedish Yorkshire), with an average parity number of 3.4+/-0.7 (mean+/-S.D.) were used. Uterine samples from the mesometrial side of both horns, taken immediately after slaughter at late dioestrus (day 17, n=3), prooestrus (day 19, n=3), oestrus (day 1, n=3), early dioestrus (day 4, n=3) and dioestrus (days 11-12, n=3), were stored in a freezer at -70 degrees C until analysed by immunohistochemistry with an avidin-biotin peroxidase method using monoclonal antibodies to lymphocyte subpopulations and MHC class II molecules. The surface and glandular epithelium as well as connective tissue layers in subepithelial and glandular areas were examined by light microscopy.For the T lymphocyte subpopulations, all oestrous cycle stages and different tissue layers taken together, the most commonly observed cell type was CD2+ cells. The largest number of CD2+ cells within the surface and glandular epithelium were observed at oestrus and early dioestrus. In the surface epithelium, a larger number of CD8+ cells compared with CD4+ cells were observed and no CD4+ cells were found within the glandular epithelium at any stage of the oestrous cycle.In the subepithelial and glandular connective tissue layers, during the oestrus cycle stages, a larger number of CD4+ cells compared with CD8+ cells were found.Endothelial cells in the connective tissue generally expressed MHC class II. However, no obvious differences between oestrous cycle stages were observed. For other cells than endothelial cells, the result was as follows. In the surface epithelium, a large number of MHC class II expressing cells was observed at oestrus compared with the other stages. No MHC class II expressing cells were found at late dioestrus and dioestrus. MHC class II expressing cells were also found in the glandular epithelium, and in the subepithelial and glandular connective tissue layers during all oestrous cycle stages but with no significant differences between stages.In conclusion, the present study showed a variation in the distribution of T lymphocyte subpopulations (CD2+, CD4+ and CD8+) and MHC class II expressing cells in the sow endometrium during different stages of the oestrous cycle. Also a variation between different tissue layers was found. It is suggested that helper and cytotoxic function of the immune system have primary locations in different tissue layers of the endometrium.     

Corrigendum to "The sow endometrium at different stages of the oestrous cycle: studies on the distribution of CD2, CD4, CD8 and MHC class II expressing" cells. [Anim. Reprod. Sci. 68 (2001) 99-109

The aim of this study was to investigate the distribution of CD2(+), CD4(+), CD8(+) lymphocyte subpopulations and MHC class II expressing cells in the sow endometrium throughout the oestrous cycle. Fifteen crossbred multiparous sows (Swedish Landrace x Swedish Yorkshire), with an average parity number of 3.4 +/- 0.7 (mean +/- S.D.) were used. Uterine samples from the mesometrial side of both horns, taken immediately after slaughter at late dioestrus (day 17, n = 3), prooestrus (day 19, n = 3), oestrus (day 1, n = 3), early dioestrus (day 4, n = 3) and dioestrus (days 11-12, n = 3), were stored in a freezer at -70 degrees C until analysed by immunohistochemistry with an avidin-biotin peroxidase method using monoclonal antibodies to lymphocyte subpopulations and MHC class II molecules. The surface and glandular epithelium as well as connective tissue layers in subepithelial and glandular areas were examined by light microscopy. For the T lymphocyte subpopulations, all oestrous cycle stages and different tissue layers taken together, the most commonly observed cell type was CD2(+) cells. The largest number of CD2(+) cells within the surface and glandular epithelium were observed at oestrus and early dioestrus. In the surface epithelium, a larger number of CD8(+) cells compared with CD4(+) cells were observed and no CD4(+) cells were found within the glandular epithelium at any stage of the oestrous cycle. In the subepithelial and glandular connective tissue layers, during the oestrus cycle stages, a larger number of CD4(+) cells compared with CD8(+) cells were found. Endothelial cells in the connective tissue generally expressed MHC class II. However, no obvious differences between oestrous cycle stages were observed. For other cells than endothelial cells, the result was as follows. In the surface epithelium, a large number of MHC class II expressing cells was observed at oestrus compared with the other stages. No MHC class II expressing cells were found at late dioestrus and dioestrus. MHC class II expressing cells were also found in the glandular epithelium, and in the subepithelial and glandular connective tissue layers during all oestrous cycle stages but with no significant differences between stages. In conclusion, the present study showed a variation in the distribution of T lymphocyte subpopulations (CD2(+), CD4(+) and CD8(+)) and MHC class II expressing cells in the sow endometrium during different stages of the oestrous cycle. Also a variation between different tissue layers was found. It is suggested that helper and cytotoxic function of the immune system have primary locations in different tissue layers of the endometrium.     

Follicular growth, endocrine response and embryo yields in sheep superovulated with FSH after pretreatment with a single short-acting dose of GnRH antagonist

The objective of this study was to characterize follicular development, onset of oestrus and preovulatory LH surge, and in vivo embryo yields of sheep superovulated after treatment with a single dose of 1.5mg of GnRH antagonist (GnRHa). At first FSH dose, ewes treated with GnRH antagonist (n=12) showed a higher number of gonadotrophin-responsive follicles, 2-3mm, than control ewes (n=9, 13.5+/-3.8 versus 5.3+/-0.3, P<0.05). Administration of FSH increased the number of >or=4mm follicles at sponge removal in both groups (19.3+/-3.8, P<0.0005 for treated ewes and 12.7+/-5.4, P<0.01 for controls). Thereafter, a 25% of the GnRHa-treated sheep did not show oestrous behaviour whilst none control sheep failed (P=0.06). The preovulatory LH surge was detected in an 88.9% of control ewes and 66.7% of GnRHa-treated sheep. A 77.8% of control females showed ovulation with a mean of 9.6+/-0.9 CL and 3.3+/-0.7 viable embryos, while ewes treated with GnRHa and showing an LH surge exhibited a bimodal distribution of response; 50% showed no ovulatory response and 50% superovulated with a mean of 12.2+/-1.1 CL and 7.3+/-1.1 viable embryos. In conclusion, a single dose of GnRHa enhances the number of gonadotrophin-dependent follicles able to grow to preovulatory sizes in response to an FSH supply. However, LH secretion may be altered in some females, which can affect the preovulatory LH surge and/or can weak the terminal maturation of ovulatory follicles.     

Temporal relationships among LH, estradiol, and follicle vascularization preceding the first compared with later ovulations during the year in mares

Diameter of the preovulatory follicle, plasma concentrations of LH and estradiol, and vascularization of the follicle wall, based on color-Doppler signals, were characterized in 40 pony mares for 6 days preceding ovulation (Days -6 to -1; preovulatory period). Comparisons between the preovulatory periods preceding the first compared with a later ovulation during the year were used to study the relationships between LH and estradiol and between vascularization and estradiol. Diameter of the preovulatory follicle was greater (P<0.02) and concentration of LH was less (P<0.02) during the first preovulatory period, whereas concentration of estradiol was not different between the first and second preovulatory periods. Vascularized area (cm(2)) of the follicle wall increased at a reduced rate during the first preovulatory period, as indicated by an interaction (P<0.03) between day and group. Vascularized area was similar between the preovulatory groups on Day -6, and a reduced rate of increase resulted in a lesser (P<0.001) area on Day -1 before the first ovulation (1.4+/-0.1cm(2)) than before a later ovulation (2.2+/-0.2 cm(2)). Results demonstrated that follicle vascularization and the LH surge were attenuated preceding the first ovulation of the year with no indication that estradiol was involved in the differences between the first and later ovulations.     

Presence of males affects the incidence of ovulation after pouch young removal in brushtail possums (Trichosurus vulpecula)

The traditional method for inducing and synchronising oestrus in the brushtail possum (Trichosurus vulpecula) is by removal of their suckling pouch young (RPY). However, our studies have recently shown that, in addition to wide variation between animals in the time of ovulation after RPY, a proportion of animals failed to ovulate. Evidence from several mammalian species indicates that the presence of males can stimulate ovarian activity and synchronise oestrus in females. The aim of this study was to determine the effect of the male on the oestrous cycle of the female brushtail possum after RPY. A total of 67 adult female brushtail possums were treated as three replicates. In order to observe the day of preovulatory follicle emergence and ovulation, animals underwent laparoscopic examination at 1–4 day intervals over a period from 0–21 days after RPY. The first replicate (N=18, May/June 1995) involved only animals kept in isolation from males, whereas the two remaining replicates compared ovarian responses between animals kept with (N=10, July 1995; N=14, June 1996) or in isolation from (N=10, July 1995; N=15, June 1996) males. The incidence of ovulation after RPY was significantly higher in females that were housed with males than in those kept in isolation from males (100%, 92.8% vs. 50.0%, 66.7%, 14.3%; P<0.001). Every animal that ovulated, had previously had a preovulatory follicle present at the site where the corpus luteum formed. Conversely, none of the animals that failed to ovulate, developed a preovulatory follicle during the period of study. The range of mean day of preovulatory follicle emergence (6.00–6.86 days), of ovulation (11.80–12.20 days) and the synchrony of ovulation between animals (range 8–17 days) after RPY, were not significantly affected by the presence of males. This study demonstrates for the first time, that the presence of males significantly increases the incidence of ovulation after RPY in the brushtail possum. However neither the timing of reproductive events nor the synchrony of ovulation were affected by presence of the male.     

Effect of oestrus synchronization methods on oestrus behaviour, timing of ovulation and pregnancy rate during the breeding and low breeding seasons in Nili-Ravi buffaloes

The objective of this study was to determine the effect of oestrous synchronization methods on oestrous behaviour, timing of ovulation and pregnancy rate during the breeding and low breeding seasons in Nili-Ravi buffaloes. In Experiment 1, oestrous behaviour and timing of ovulation were determined from (n=34) oestruses. The mean (+/- S.E.M.) time of ovulation after the onset of standing oestrus was greater (P<0.05) in PGF(2alpha)-induced luteolysis (30.6+/-1.5h) compared to Ovsynch buffaloes (15.0+/-0.8h). In Experiment 2, pregnancy rates were compared between two methods of synchronization (detected oestrus and Ovsynch protocol) during the breeding and low breeding seasons. Pregnancy rates of buffaloes bred at detected oestrus (62.5%) or by the Ovsynch protocol (36.3%) during the breeding season did not differ significantly (P>0.05) from those which were inseminated during the low breeding season (55.5%) and (30.4%), respectively. This study demonstrates clearly that (1) timing of ovulation in Nili-Ravi buffalo is about 30h after the onset of standing oestrus and (2) buffaloes can be successfully synchronized with optimum fertility using either PGF(2alpha) alone (detected oestrus) or using (Ovsynch protocol) during low breeding season, to calve during the period when milk availability is short.     

Follicular dynamics and temporal relationships among body temperature, oestrus, the surge of luteinizing hormone and ovulation in Holstein heifers treated with norgestomet

The effects of chronic treatment with norgestomet on follicular dynamics, corpus luteum growth and function as well as the temporal relationships among body temperature, oestrous behaviour, the luteinizing hormone (LH) surge and ovulation following implant removal were studied in 16 Holstein heifers. Oestrous cycles of the heifers were initially synchronized using 2 injections of prostaglandin F-2 alpha (PGF-2 alpha) 12 days apart. The heifers were then implanted with a norgestomet ear implant for 9 days, beginning either at the middle of the synchronized cycle (dioestrus) or at the end of the synchronized cycle (pro-oestrus). Follicular dynamics, corpus luteum growth and regression, and plasma progesterone were not affected by norgestomet treatment at dioestrus. The dominant follicle present at the time of norgestomet implantation in the pro-oestrus group was maintained during the 9-day implant period of 6 of 8 heifers and ovulated after implant removal. Time from implant removal to onset of standing oestrus and time to LH peak following implant removal were highly correlated with the time of ovulation (r = 0.92 and 0.96, respectively). Onset of standing oestrus and the LH peak and the onset of standing oestrus and peak vaginal and rectal temperatures were also highly correlated (r = 0.96, 0.82 and 0.81, respectively). It is concluded that any decrease in pregnancy rates following treatment with norgestomet is not due to asynchrony among oestrus, the LH surge and ovulation.     

Analyses of the variation in the interval from an injection of prostaglandin F2α to oestrus as a method of studying patterns of follicle development during dioestrus in dairy cows

Lactating dairy cows experiencing normal oestrous cycles were injected once with either 0.5 mg of an analogue of prostaglandin F_2α (PGF) (Cloprostenol, 435 cows) or 25 mg of a PGF-Tham salt (237 cows) when at days 7–16 of the cycle (oestrus = day 0). In these two trials, 91% and 93% of the cows were detected in oestrus from 3–10 days post-injection increasing from 81% to 98% with advancing dioestrus. Over 70% of detected cows injected on day 7 (early dioestrus) or day 16 (late dioestrus) were in oestrus from 48 to 72 h post-injection. Comparable response rates among cows injected on days 11 and 12 (mid-dioestrus) were less than 30% with most response intervals being at 4 and 5 days post-injection (73 h–120 h). The variability in response intervals generally decreased with advancing dioestrus. A regression model for ordinal data, with post-injection interval to oestrus as the ordinal response and stage of cycle at injection as the explanatory variable, showed that both the interval to oestrus and the variation in this interval varied with the stage of cycle at injection.These response intervals appear to reflect a wave-like pattern in ovarian follicle development during dioestrus. The probability of the presence of a follicle in a less advanced stage of development at the time of PGF injection is greatest among animals treated during mid-dioestrus.     

The effect of melatonin implants on the response to the male effect and on the subsequent cyclicity of Rasa Aragonesa ewes implanted in April

Rasa Aragonesa ewes were used to evalutate whether treatment with melatonin implants in spring could modify: (i) the response to the male effect in terms of oestrous behaviour and ovulation rate; and (ii) the maintenance of sexual activity and ovulation rate at medium term, i.e. over the next 306 days. On 12 April, 42 ewes were divided into two groups, with (M; n = 21) or without (C; n = 21) a subcutaneous implant containing 18 mg melatonin. On 17 May (day 0), three aproned rams were introduced to each group to induce a ram effect. Ewes were observed for oestrus daily. The rams were removed 40 days later after which one aproned ram was introduced daily. Oestrous detection continued until 28 February, 306 days after the first male-female contact. The ovulation rate was determined by endoscopy in the first three cycles after ram introduction and in September-October and January. Progesterone was assayed from blood samples taken on 6 May, 10 and from day 0 to day 22 after ram introduction. Luteal activity before ram introduction was seen in 33% (M) and 29 (C)% of the ewes, respectively. Significantly more M ewes showed oestrous behaviour during the first 40 days after ram introduction (M: 100%; C: 62%; P < 0.01). Similar differences were observed for ewes anovulatory at ram introduction (M: 100%, C: 47%; P < 0.01). These differences were maintained over the three oestrous cycles in both groups. Treatment with melatonin implants was without detrimental effect on cyclic functions in the following breeding season, after seasonal anoestrus. Melatonin treatment significantly increased (P < 0.05) the mean ovulation rate of the first (1.62 +/- 0.11 versus 1.31 +/- 0.13), second (1.78 +/- 0.15 versus 1.36 +/- 0.15) and third cycles (M: 1.73 +/- 0.12 versus C: 1.27 +/- 0.14). There was a significant interaction between the effects of cyclicity at day 0 and melatonin treatment on the ovulation rate in the first cycle (P < 0.05). The mean ovulation rates of both groups were similar at the beginning (September) and middle (January) of the subsequent breeding season. Overall, the results confirmed that melatonin implants, combined with the ram effect, improved the reproductive parameters of reduced-seasonality ewes during spring mating, without impairing sexual activity or ovulation rate during the subsequent breeding season.     

Origin and fate of preovulatory follicles after induced luteolysis at different stages of the luteal phase of the oestrous cycle in goats

The effect of day of induced luteolysis on follicle dynamics, oestrus behaviour and ovulatory response in goats was studied by administering cloprostenol on Day 5 (n=10), Day 11 (n=10), or Day 16 (n=10) after detection of oestrus. Stage of the luteal phase affected the interval from cloprostenol injection to onset of oestrus, with behavioural oestrus being observed earlier in goats treated early in the luteal phase (43.4+/-3.2 h on Day 5 versus 57.0+/-2.6 h on Day 11 and 56.7+/-2.7 h on Day 16, P<0.01). The group treated on Day 5 also tended to have a higher proportion of does which exhibited oestrus behaviour (P=0.07) and ovulation (P=0.06). In all the cycles, at least one of the ovulatory follicles arose from antral follicles present in the ovary at cloprostenol injection. In 66.7% of monovular cycles, the ovulatory follicle was the largest follicle on the day of luteolysis. In 33.3% of polyovulatory cycles, one of the ovulatory follicles was the largest one present when cloprostenol was administered. In 80% of polyovulatory cycles, the second ovulatory follicle was present on the day of luteolysis; but in the three remaining cycles, the second ovulatory follicle emerged later. This shows that the largest follicle may not exert dominance over other follicles in the goat. Evaluation of follicular dynamics in different phases of luteal activity in current experiment showed an attenuation of dominance in the mid-luteal period. In does treated early or late in the luteal phase, the number of new growing follicles decreased with time (P<0.01 and 0.05, respectively), the mean number of follicles reaching 4-5mm in size also decreased (P<0.001 and 0.01, respectively) and the number of regressing follicles increased (P<0.05). These effects did not reach statistical significance in does treated in the mid-luteal phase.     

Synchronisation of oestrus in dairy cows using prostaglandin F2alpha,gonadotrophin-releasing hormone,and oestradiol cypionate

An oestrous synchronisation protocol was developed for use in lactating dairy cows using PGF(2alpha), GnRH, and oestradiol cypionate (ECP). In experiment 1, lactating dairy cows received two injections of PGF(2alpha) (on days 0 and 11) (PP; n=10) or two injections of PGF(2alpha) (days 0 and 11) and 100 microg of GnRH on day 3 (PGP; n=10). In experiment 2, cows were treated with PGP (n=7), or PGP and 1 mg of ECP at the same time (PGPE(0); n=7) or 1 day after the second PGF(2alpha) injection (PGPE(1); n=7). In experiment 3, 101 lactating dairy cows in a commercial herd were assigned to one of three treatments; PP, PGP, or PGPE(1). Follicular growth was measured by ultrasound in experiments 1 and 2. Every cow (experiments 1, 2, and 3) was blood sampled at selected intervals for progesterone and oestradiol assays and inseminated at oestrus. In experiment 1, a higher percentage of GnRH-treated cows ovulated after the first PGF(2alpha) injection (90% versus 50%; P<0.05). The GnRH-treated cows tended to have a larger dominant follicle present at the time of the second PGF(2alpha) injection (16.5+/-0.5 mm versus 15.0+/-0.7 mm; P<0.10). The percentage of cows that ovulated after the second PGF(2alpha) injection was similar (60%). In experiment 2, cows treated with ECP had higher peak preovulatory concentrations of oestradiol in plasma (6.99+/-0.63 versus 3.63+/-0.63; P<0.01) following the second PGF(2alpha) injection and a higher percentage ovulated (86% versus 43%; P<0.05). A higher percentage of PGPE(1)-treated cows in experiment 3 were observed in standing oestrus and ovulated after the second PGF(2alpha) injection (standing oestrus, 26.4, 34.3, and 62.6%, P<0.01; ovulated, 56, 63, and 78%, P<0.05; PP, PGP, and PGPE(1), respectively). In conclusion, the PGP protocol increased the number of cows that ovulated after the first PGF(2alpha) injection and produced a more mature dominant follicle at the time of the second PGF(2alpha) injection. Adding ECP to PGP (PGPE(1)) enhanced the expression of oestrus and increased ovulation percentage. The combination of PGP and ECP is potentially a new method to routinely synchronise oestrus and ovulation in dairy cows.