Regular and irregular characteristics of ovulation and the interovulatory interval in mares

Characteristics of regular and irregular ovarian events were studied by daily ultrasound examination in 102 non-bred mares. The mean diameter of the prevoluatory follicle on the day before a single ovulation was significantly higher in April (46 mm) and May (48 mm) than in July (40 mm). Diameter was also significantly different among single (44 mm), double unilateral (35 mm), and double bilateral (40 mm) preovulatory follicles. The incidence of apparent anovulation within the ovulatory season was 4.7%. All apparent anovulations involved filling of the follicle with blood (hemorrhagic follicles). The incidence of diestrous ovulations was 4%. Prolonged interovulatory intervals occurred in 6 of 69 (9%) intervals. In 2 intervals (30 and 33 days), both the luteal and follicular phases were prolonged and no structural ovarian or uterine irregularities were detected. Three prolonged intervals (34, 41, and 49 days) were associated with hemorrhagic follicles and 1 was associated with a late diestrous ovulation (47 days). The condition known as spontaneous persistence of the corpus luteum, which is thought to be common in nonpregnant mares, was not detected in any of 69 interovulatory intervals.     

Effects of low-dose follicle-stimulating hormone administration on follicular dynamics and preovulatory follicle characteristics in dairy cows during the summer

The well-documented phenomenon of reduced conception rate in dairy cows during the hot season involves impaired functioning of the ovarian follicles and their enclosed oocytes. Three experiments were performed to examine the administration of low doses of follicle-stimulating hormone (FSH) to induce turnover of follicles that are damaged upon summer thermal stress and to examine whether this FSH administration has beneficial effects on preovulatory follicles. In experiment 1, synchronized heifers were treated with 100 mg of Folltropin-V (n = 7) or 4.4 mg of Ovagen (n = 6) on day 3 of the estrous cycle. Treatment with both FSH sources resulted in greater (P < 0.05) numbers of follicles than in control animals (n = 12) on day 6 of the estrous cycle, indicating that low doses of FSH can increase the number of emerging follicles in a follicular wave. In experiment 2, milking cows were assigned to a control group (n = 4) or treated with 2.2 mg (FSH-2.2; n = 6) or 4.4 mg (FSH-4.4; n = 5) Ovagen. Follicle-stimulating hormone was administrated on day 3 or 4 and day 10 or 11 of the estrous cycle, coinciding with emergence of the first and second follicular waves, respectively. The number of follicles emerging during the first wave tended to be higher (P < 0.1) in FSH-4.4-treated cows than in controls. The second-wave dominant follicles emerged 2 d later in the treated cows and were smaller in diameter (P < 0.05) than controls, 2 d before aspiration. Despite being younger, the preovulatory follicles of FSH-4.4 cows expressed a steroidogenic capacity that was similar to controls with a tendency toward greater insulin concentrations (P < 0.09). In experiment 3, milking cows were assigned to a control group (n = 6) or treated with 4.4 mg Ovagen (FSH-4.4; n = 6). Follicle-stimulating hormone was administrated on day 3 and day 12 or 13 of the estrous cycle. The number of emerging follicles was higher (P < 0.05) in the treated vs control cows. However, the features of the preovulatory follicle developed in the subsequent cycle did not differ between groups. In summary, low doses of FSH can efficiently induce follicular turnover accompanied by a modest effect on the preovulatory follicle of the treated cycle. It appears that the administration of low doses of FSH, precisely timed to synchronize with the emergence of follicular waves, might have a beneficial effect on the preovulatory follicle and its enclosed oocyte.     

Detection of chemotactic factors in preovulatory follicular fluid from mares

Ovulation has been likened to an inflammatory process. Inflammatory cells accumulate in the ovulating follicle, presumably because of chemotactic factors. Chemotactic activity was measured in fluid aspirated from follicles of estrous mares 0, 12, 24, and 36 hours after ultrasonographic detection of a 35-mm follicle and IV treatment with 2,500 IU of human chorionic gonadotropin. Chemotaxis was assessed by measuring directional migration of equine neutrophils under agarose. Follicular fluid acted as a chemoattractant for neutrophils, but there was no significant difference in chemotactic activity among different time intervals after administration of human chorionic gonadotropin. On the basis of results of various treatments, chemotactic properties of serum and follicular fluid were similar. Chemotactic activity was significantly reduced by heating (56 C for 30 minutes) and by trypsinization and was virtually removed by charcoal treatment. Dialyzing the follicular fluid (3,500 and 8,000 molecular weight cut-off) significantly reduced the chemotactic activity of follicular fluid and serum. The importance of chemotactic factors in the process of ovulation in the mare is yet to be established.     

FSH and LH concentrations preceding post-partum ovulation in the mare

Equine follicle stimulating hormone (FSH) and luteinizing hormone (LH) were measured in the serum obtained from pregnant mares, bled daily, from up to 17 days before parturition until the first ovulation after parturition. LH was at baseline levels until 2 days before ovulation when it started, and continued, to rise until after ovulation. FSH surges occurred at approximately 24 and 14 days before this post-partum ovulation (12 and 2 days before parturition), thus showing a similar pattern to the cyclic mare, consistent with the hypothesis that 2 surges at these times prepare and prime follicles for subsequent ovulation. The high plasma oestradiol levels that occurred during and immediately before these FSH surges did not show a negative feedback, or the inverse relationship between FSH and oestradiol which occurs in the cyclic mare, and in other species.     

Milk and serum progesterone levels in mares after ovulation

Twenty-four Finnhorse mares were examined by rectal palpation and ultrasonography every 6 h during late oestrus to determine the time of ovulation. Milk and serum samples were collected every 6 h after the detected ovulation for progesterone analysis. The progesterone rises took place within 0-54 h and 0-60 h after ovulation, in milk and serum, respectively. Statistically significant differences (p less than 0.05) in progesterone levels were observed for the first time 12-18 h and 18-24 h after ovulation, in serum and milk, respectively, as compared to progesterone levels 0-6 h after ovulation.     

Colic-like discomfort associated with ovulation in two mares

Discomfort manifested by colic-like clinical signs in 2 young mares was presumed to be attributable to ovarian pain associated with follicular enlargement and ovulation. Diagnosis was based on the lack of detectable evidence of gastrointestinal disease, the finding of a large ovarian follicle or recent ovulation, the repetition of signs during several subsequent estrual periods, and the clinical response to pharmacologic suppression of estrus and ovulation. The similarity of the clinical signs in these 2 mares to cyclic intermenstrual pain in women was considered.     

Natriuretic peptide system regulation in granulosa cells during follicle deviation and ovulation in cattle

Natriuretic peptides (NPs) are known to regulate reproductive events in polyovulatory species, but their function and regulation in monovulatory species remain to be fully characterized. Using a well‐established in vivo model, we found that bovine granulosa cells from follicles near the deviation stage express mRNA for the three NP receptors (NPR1, NPR2 and NPR3), but not for NP precursors (NPPA, NPPB and NPPC). The abundance of NPR3 mRNA was higher in dominant compared to subordinate follicles at the expected time of follicular deviation. After deviation, mRNA for all NP receptors was significantly more abundant in the dominant follicle. Intrafollicular inhibition of oestrogen receptors downregulated NPR1 mRNA in dominant follicles. In granulosa cells from preovulatory follicles, NPPC mRNA increased at 3 and 6 h after systemic GnRH treatment, but decreased at 12 and 24 h to similar levels observed in samples collected at 0 h. After GnRH treatment, NPR1 mRNA was upregulated at 24 h, NPR3 mRNA gradually decreased after 3 h, while NPR2 mRNA was not regulated. The mRNA expression of the enzyme FURIN increased at 24 h after GnRH treatment. These findings revealed that the expression of mRNA encoding important components of the NP system is regulated in bovine granulosa cells during follicular deviation and in response to GnRH treatment, which suggests a role of NP system in the modulation of these processes in monovulatory species.     

Characterization of antral follicle populations during the estrous cycle in pigs selected for ovulation rate

The objectives of this study were to characterize and compare ovarian follicular populations in Gene Pool Control (GPC, randomly selected) and Relax Select line (RS, nine generations of selection for high ovulation rate followed by six generations of random selection) gilts during different stages of the estrous cycle. Thirty-five RS and 23 GPC gilts were allotted randomly within litter for ovary recovery on either d 3, 15 or 19 of the estrous cycle. Surface follicles on the ovaries were classified by size (small, less than 3 mm; medium, 3 to 6.9 mm; large, 7 to 12 mm), and counts were recorded for each ovary. Ovarian weight (OW), number of corpora lutea (CL), follicular fluid volume (FFV) from small, medium and large follicles, residual ovarian weight and follicular fluid weight (FFW) also were recorded. Total numbers of small and medium follicles were greatest on d 15, whereas total number of large follicles and FFW were greatest on d 19. The OW, FFW and follicle numbers of all classes were lowest on d 3. The RS gilts expressed longer interestrous intervals (21.9 vs 20.4 d, P less than .05) and higher ovulation rates (18.5 vs 15.3 CL, P less than .01) than GPC gilts. The left ovary of RS gilts was responsible for most of the ovulation rate advantage (10.3 vs 7.4 CL, P less than .01) Overall, GPC gilts had more total small follicles than RS gilts (P less than .01). The advantage was due primarily to higher numbers of small follicles at d 15.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of interrupted photoperiods on the induction of ovulation in anestrous mares

The ability of interrupted photoperiods to induce early estrus and ovulation was examined. Horse mares were exposed to long (16 h light) or short (10 h light), noninterrupted photoperiods, ambient light, or various interrupted photoperiod treatments from December 1 to April 15 (135 d). Follicular development was assessed by rectal palpation and estrous behavior was determined by teasing with a stallion. Serum concentrations of progesterone were used as an indicator of corpus luteum function. Differences among the light treatment groups were compared for the following behavioral and ovarian characteristics: days to first detectable 3-cm follicle, days to first estrous behavior, days to first ovulation, the number of mares ovulating within the treatment period, and the number of ovulations within the treatment period per mare. Compared with the ambient and 10L:14D (L = h of light and D = h of darkness) photoperiod treatments, ovulation was advanced to the greatest extent by a photoperiod of 16L:8D and the interrupted photoperiod 10L:8D:2L:4D. These two stimulatory photoperiod treatments were characterized by the presence of light 8 to 10 h after dusk. Therefore, the present data are consistent with an external coincidence model for the induction of seasonal breeding in horses, with the photoinducible phase occurring within the period 8 to 10 h after dusk.     

Use of gonadotropin-releasing hormone for hastening ovulation in transitional mares

Natural GnRH and its analog have potential for hastening ovulation in mares. A study was conducted to evaluate the efficacy of a GnRH agonist given either as an injectable or s.c. implant for induction of ovulation in mares. Forty-five seasonally anestrous mares (March) were assigned to one of three groups (n = 15/group): 1) untreated controls; 2) i.m. injection of the GnRH agonist buserelin at 12-h intervals (40 micrograms/injection for 28 d or until ovulation) and 3) GnRH agonist administered as a s.c. implant (approximately 100 micrograms/24 h for 28 d). Six mares per group were bled on d 0, 7, 14 and 21 after injection or insertion of implant. Samples were taken at -1, -.5 and 0 h and at .5, 1, 1.5, 2, 4, 6 and 8 h after GnRH. Additional daily samples were drawn for 28 d after injection or until ovulation. Samples were assayed for concentration of LH and FSH. Progesterone concentrations were determined in samples collected on d 4, 6 and 10 after ovulation. Number and size of follicles and detection of ovulation were determined by ultrasonography. Number of mares induced to ovulate within 30 d was 0 of 15, 7 of 15 and 9 of 15 for groups 1, 2 and 3, respectively. During treatment, follicle sizes were smaller for mares in group 3 (implant). The LH response to GnRH agonist (area under curve) was similar among groups at d 0 but was greater (P less than .05) for mares in group 3 on d 7 and 14 and groups 2 and 3 on d 21 than for controls. A similar pattern was detected for peak concentrations of LH after GnRH on d 0, 7, 14 and 21. Daily concentrations of LH remained low in untreated control mares compared with GnRH-treated mares throughout the sampling period. Concentrations of LH for mares in group 3 that ovulated were elevated greatly above those for group 2 mares, whereas concentrations of FSH were similar in both treatment groups prior to ovulation.     

Serrated granulosa and other discrete ultrasound indicators of impending ovulation in mares

The incidence of discrete structural changes in the preovulatory follicle as ovulation approaches was studied sequentially by ultrasonography. Examinations were done every 12 hours in 27 mares, beginning when the follicle was ≥35 mm. The following discrete end points were recorded as present or absent: 1) serration of granulosa, indicated by an irregular or notched appearance; 2) decreased turgidity, indicated during transducer pressure; 3) loss of spherical shape; 4) an apex, indicated by a reduced area at one end; and 5) echoic spots floating in the antrum. When the records were examined as though scanning had been done only every 24 hours beginning at 35 mm, distinct serration was detected at the examination before ovulation in 37% of mares but not earlier. When mares were examined every 12 hours, serration was detected at the last examination in 59%. Decreased turgidity at the last 12-hour examination was detected concomitantly with serration, but was detected alone in 9% to 12% of previous examinations. Mares with serration at the last examination at 12-hour intervals were examined every hour thereafter (n = 14). Serration and decreased turgidity were present at each examination until ovulation 4.9 ± 0.7 hours later. Loss of spherical shape initially occurred less frequently than decreased turgidity, but the incidence increased from 50% to 100% during 6 to 1 hours before ovulation. The incidence of an apical area reached 100%, and echoic spots increased to 50% during the few hours before ovulation. Results indicated that serrated granulosa and the other discrete indicators were useful for predicting impending ovulation; however, optimal efficiency would require examinations every few hours.     

Ovulation of the preovulatory follicle originating from the first-wave dominant follicle leads to formation of an active corpus luteum

The objective of our study was to compare the characteristics of the corpus luteum (CL) formed after ovulation of the dominant follicle (DF) of the first follicular wave (W1) and those of the CL formed after ovulation of the DF of the second (induced) follicular wave (W2). Non-lactating Holstein cows were used for this study. In Experiment 1, cows were treated with PGF2α and GnRH on days 6 and 8 (day 0 = day of follicular wave emergence) for W1 (n = 6) and W2 (n = 6), respectively. Dominant follicles were aspirated on day 9 to quantify the amounts of mRNA (VEGF120, VEGF164, FGF-2, StAR, P450-scc and 3β-HSD) in granulosa cells (GC). In Experiment 2, the size and blood flow area of the CL formed after ovulation of the DF in W1 (W1CL; n = 6) and W2 (W2CL; n = 6) (the day of DF ovulation in W1 and W2 was day 10) were evaluated on days 12, 15, 18 and 21. The plasma P4 concentration was measured on days 10 to 21. The amounts of VEGF164, P450-scc and 3β-HSD mRNA were higher (P < 0.05) in the DF in W1, and those of VEGF120,FGF-2 and StAR mRNA tended to be higher (P < 0.1) in the DF in W1. The size of the CL was greater in the W1CL on days 15, 18 and 21. The blood flow area of the CL was greater in the W1CL on days 12 and 15. The plasma P4 concentrations were higher in the W1CL. These results indicate that the CL formed after ovulation of the DF in W1 was greater in terms of size, blood flow and plasma P4 concentration.     

Efficacy of altrenogest administration to postpone ovulation and subsequent fertility in mares

A recent report suggested administration of altrenogest during the follicular phase could postpone ovulation. Based on these results, two questions were generated. We first hypothesized that by initiating a altrenogest treatment earlier in the estrous cycle, a greater and/or more consistent delay in ovulation would result. Second, we hypothesized that exposure to elevated progestin concentrations might alter viability of the ovulatory follicle and oocyte. The focus of the first experiment was to determine if initiation of altrenogest treatment at different stages of the estrous cycle would yield a more predictable time to ovulation, whereas the second experiment was designed to determine whether mares receiving altrenogest during estrus had compromised fertility. In the first experiment thirty mares of mixed light breed, ranging in age from 5-15 years, were randomly assigned to one of three groups. The two treated groups received altrenogest (0.088 mg/kg of body weight) for two days once a follicle of 30 or 35 mm in diameter was detected. Control mares were not treated. Mares treated with altrenogest whether initiated at the detection of a 30 or 35 mm follicle demonstrated similar (P>.05) day to ovulation interval when adjusted to 35 mm (5.4 and 5.6 days, respectively). Both treated groups demonstrated a delayed interval (P<.05) when compared to control (3.9 days). Thirty-six mares of similar breed and age, were randomly assigned to two groups for use in the second experiment. All mares were monitored daily via transrectal ultrasonography from the time a 35 mm or greater follicle was detected until ovulation. Treated mares received daily doses of altrenogest (0.088 mg/kg of body weight) for two days once a follicle of 35 mm or greater was detected. Control mares received no treatment. Fertility data were collected from mares inseminated every other day with 500 million motile spermatozoa from one of two stallions with proven fertility. Pregnancy data were collected via transrectal ultrasonography at days 12, 14 and 16 post-ovulation. Ovulation data were collected from 27 control cycles and 26 treated cycles. Contrary to previous reports and Experiment 1, no difference (P=0.35) was noted between groups with respect to days to ovulation. Control mares averaged 4.14 days and treated mares averaged 4.7 days to ovulation from initial detection of a 35 mm follicle. Fertility data were also similar (P=0.8) between control and treated mares (66.6% and 61.5% per cycle, respectively). Interestingly, a greater number (P=0.017) of treated cycles (5/26) resulted in follicular regression than did control cycles (0/27). While these data suggest that this dosage of altrenogest may not postpone ovulation, it did appear related to increased incidence of follicular regression. Fertility was unaffected, however, in those mares that ovulated. Further studies are needed in which initiation at different stages of estrus and different doses of altrenogest are used.     

Efficacy of short-term administration of altrenogest to postpone ovulation in mares

Estrogen from a growing follicle stimulates the preovulatory surge of luteinizing hormone (LH) while progesterone (P) is known to suppress LH. The possibility exists that administration of P, in the presence of an ovulatory follicle, would sufficiently suppress LH and, therefore, delay ovulation. The objective of this research was to elucidate the potential for oral administration of altrenogest (17-Allyl-17β-hydroxyestra-4,9,11-trien-3-one) to postpone ovulation of a preovulatory follicle (35 mm) for approximately two days. Fourteen light-horse mares, ranging in age from two to 19 years, were randomly assigned to one of three treatments (A-.044 mg/kg BW altrenogest for two days; B-.088 mg/kg BW altrenogest for two days; and C- no altrenogest). Mares began treatment when a 35-mm or greater follicle was observed via real-time transrectal ultrasonography. Both number of days until ovulation and follicular maintenance differed between treated and control mares. Number of days until ovulation was increased (P<.05) for mares in treatment A when compared with the control mares. Follicular diameter maintenance, a measurement of follicular diameter throughout treatment, also increased (P<.05) for mares in treatment A when compared with the control mares. Mean LH concentration was not different between mares treated with altrenogest at either treatment dose when compared with the control mares. Pregnancy rates and embryonic vesicle size change were also measured to determine potential effects of altrenogest administration. No differences (P>.05) were found in either characteristic.Short-term administration of altrenogest increased the number of days to ovulation. Further study is warranted to prove conclusively that altrenogest increases follicular maintenance, alters the preovulatory LH surge, and has no detrimental effects upon reproductive efficiency.