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Studies on a local effect of boar seminal plasma on ovulation time in gilts.

Previous studies showed that intrauterine infusion of seminal plasma at the onset of oestrus could advance ovulation in pigs, possibly to enhance the chances of fertilization by optimizing the chronological events of fertilization. This effect has been attributed to a local unilateral mechanism whereby infusion into a single uterine horn advances ovulation in the adjacent ovary. The present study was designed to elucidate possible mechanisms of local signal transduction. In a series of five experiments using 43 gilts, the ovarian response was investigated after infusion of seminal plasma at different sites of the female reproductive tract. The time of ovulation was detected sonographically at 4- or 2-h intervals. Single uterine horn infusion of 100 ml seminal plasma advanced ovulation on the ipsilateral ovary by 9.3 h (mean) compared with the contralateral ovary. Dissection of the ipsilateral isthmus abolished the unilateral seminal plasma effect. Unilateral infusion of 50 microliters or 1 ml seminal plasma or 50 microliters of the concentrated 1-10 kDa fraction in the lower isthmus was ineffective. Application of 5 ml seminal plasma into the tip of a ligated uterine horn lead to 3.6 h (mean) earlier ovulation on the adjacent ovary. In contrast, the infusion of 5 ml NaCl showed no effect. Application of 5 ml seminal plasma in the middle of the uterine horn between two ligatures was ineffective. It is concluded that, for the transduction of the local signal involved in the advancement of ovulation, contact of seminal plasma with the epithelium of the utero-tubal junction is essential.

Animals↗

Consistency and variation in fertility investigations in Europe.

A questionnaire survey among the teaching departments of obstetrics and gynaecology in Western Europe (response rate 71%) revealed only weak adherence to the World Health Organization recommendations for the standard investigation of the infertile couple. Both general and specific examinations were applied more frequently in the female than in the male partner. Although semen analysis and the ascertainment of ovulation were standard in virtually all departments, the criteria for normal semen and the methods used for the detection of ovulation varied greatly among both departments and countries. There were also large differences among countries in the preferred standard method for testing tubal patency. The data suggest that fertility investigations are based more on tradition and personal preferences than on the demonstrated utility of its components.

Europe↗

A statistical approach to the determination of the fertile period.

17 beta-Estradiol (E2), basal body temperature (BBT) and luteinizing hormone (LH), were measured in normally menstruating women. The preovulatory E2 observations were characterized by variations about a constant level during the preovula-peak. The BBT observations typically varied about a constant level during the preovulatory phase then about a higher constant level during the postovulatory phase. The change points in E2 and BBT were detected using a Bayesian detection rule and a midcycle 'fertile period' was marked off. The midcycle 'fertile periods' appear to bracket the ovulation event and the method shows promise as a natural family planning method. Several practical limitations of the method are discussed.

Bayes Theorem↗

Influence of time of insemination relative to ovulation and frequency of insemination on gilt fertility.

The objectives of this study were to determine the optimal time of insemination in the pre-ovulatory period (from 32 to 0 h before ovulation) and to evaluate once-daily versus twice-daily inseminations in gilts. In Experiment 1, pre-puberal gilts (n=102) were observed for estrus every 8h and ultrasonography was performed every 8h from the onset of estrus to confirmation of ovulation. The gilts were inseminated once with 4 x 10(9) spermatozoa at various intervals prior to ovulation. Pregnancy detection was conducted 24 days after AI and gilts were slaughtered 4-6 days later. Corpora lutea and the number of viable embryos were counted and the embryo recovery rate was calculated (based on the percentage of corpora lutea). Inseminations performed <24h before ovulation resulted in a higher embryo recovery rate (P=0.02) and produced 2.1 more embryos (P=0.01) than inseminations >or=24h before ovulation. However, the pregnancy rate was reduced when inseminations were performed >16 h before ovulation (P=0.08). In Experiment 2, pre-puberal gilts (n=105) were observed for estrus every 12h and ultrasonography was performed every 12h from the onset of estrus to confirmation of ovulation. Gilts were inseminated (with 4 x 10(9) spermatozoa) 12h after the onset of estrus, with inseminations repeated either every 12h (twice-daily) or 24h (once-daily) during estrus. The gilts were allowed to farrow. There were no differences (between gilts bred twice-daily versus once-daily) for return to estrus rate (P=0.36) and adjusted farrowing rate (P=0.19). However, gilts inseminated once-daily had 1.2 piglets less than those inseminated twice-daily (P=0.09). In conclusion, gilts should be inseminated up to 16 h before ovulation, as intervals >16 h reduced pregnancy rate and litter size.

Animals↗

Effect of pollinator-inflicted ovule damage on floral abscission in the yucca-yucca moth mutualism: the role of mechanical and chemical factors.

The long-term persistence of obligate mutualisms (over 40 Mya in both fig/fig wasps and yucca/yucca moths) raises the question of how one species limits exploitation by the other species, even though there is selection pressure on individuals to maximize fitness. In the case of yuccas, moths serve as the plant's only pollinator, but eggs laid by the moths before pollination hatch into larvae that consume seeds. Previous studies have shown that flowers with high egg loads are more likely to abscise. This suggests that yucca flowers can select against moths that lay many eggs per flower through selective abscission of flowers; however, it is not known how yucca moths trigger floral abscission. We tested how the moth Tegeticula yuccasella triggers floral abscission during oviposition in Yucca filamentosa by examining the effects of ovipositor insertion and egg laying on ovule viability and floral abscission. Eggs are not laid at the site of ovipositor insertion: we used this separation to test whether wounded ovules were more closely associated with the ovipositor site or an egg's location. Using a tetrazolium stain to detect injured ovules, we determined whether the number of ovipositions affected the number of wounded ovules in naturally pollinated flowers. Two wounding experiments were used to test the effect of mechanical damage on the probability of floral abscission. The types of wounds in these experiments mimicked two types of oviposition-superficial oviposition in the ovary wall and oviposition into the locular cavity-that have been observed in species of Tegeticula. The effect of moth eggs on ovule viability was experimentally tested by culturing ovules in vitro, placing moth eggs on the ovules, and measuring changes in ovule viability with a tetrazolium stain. We found that ovules were physically wounded during natural oviposition. Ovules showed a visible wounding response in moth-pollinated flowers collected 7-12 h after oviposition. Exact location of wounded ovules relative to eggs and oviposition scars, as well as results from the artificial wounding experiments, showed that the moth ovipositor inflicts mechanical damage on the ovules. Significantly higher abscission rates were observed in artificially wounded flowers in which only 4-8% of the ovules were injured. Eggs did not affect ovule viability as measured by the tetrazolium stain. These results suggest that physical damage to ovules caused by ovipositing is sufficient to explain selective fruit abscission. Whether injury as a mechanism of selective abscission in yuccas is novel or a preadaptation will require further study.

Adaptation, Physiological↗

LH surge induction by GnRH agonist at the time of ovulation.

Eight women were treated for 1 cycle at the time of ovulation with GnRH agonist (3 injections of 0.2 mg buserelin s.c. at 12-hour intervals) to obtain follicular rupture. Clomiphene citrate was administered to 1 patient and pure FSH to 2 patients, whilst in the case of a woman with hypothalamic amenorrhea a pulsatile GnRH regimen was used. Four patients had an untreated follicular phase. When the maximal follicular diameter was 20-22 mm all treatments were withheld and GnRH-A was administered. Plasma LH, FSH, progesterone and estradiol were determined 24 hours before, at the time of and 12, 24 and 48 hours after the 1st injection of buserelin. Ovulation was detected in all cycles. LH levels increased dramatically from baseline levels of 14.4 +/- 4.1 to 155 +/- 48 IU/l 12 hours after the beginning of treatment, then returned to preovulatory values 48 hours later (13.0 +/- 3.9 IU/l). The duration of the luteal phase was 13.2 days and normal mid-luteal progesterone plasma levels were detected (39.8 +/- 9.3 nmol/l). These data suggest that the GnRH agonist can be successfully used at the time of ovulation to induce an endogenous ovulatory LH peak and that it can be used in conjunction with different medical treatments to induce follicular maturation.

Adult↗

[Recovery of preovulatory human oocytes (author's transl)].

Rapid radio-immune assaying of LH was carried out four times daily on 52 women in the preovulatory phase of the cycle. This method enables the precise detection of the LH surge and recovery of the mature oocyte by laparoscopic aspiration of the follicular content, just before ovulation, which is between 28 and 36 hours after the start of the LH surge. The oocytes was recovered in 27 out of 41 patients with a normal cycle (66%). The rate of recovery per follicle aspirated is comparable in a natural cycle to that after treatment with Clomid (80 to 84%).

Adult↗

Seasonal effects on attempts to synchronize estrus and ovulation by intravaginal application of progesterone-releasing device (PRID) in mares.

To investigate seasonal effects on the efficacy of estrus synchronization in mares, we administered a progesterone-releasing device (PRID) intravaginally to eight Haflinger mares for 11 days. In January 3 of 8 mares responded to the treatment with estrus and ovulation, in March 7 with estrus and 6 of 7 mares with ovulation, in June 6 of 7 and in October 7 of 8 mares with estrus and ovulation. Follicle distribution patterns at PRID insertion were different between January/October, March/June and June/October (P<0.05). Number of follicles decreased during PRID treatment in January, March and June (difference of number of follicles at Day 12 minus number of follicles at Day 1: -4.2+/-2.7, -0.9+/-0.9 and -4.9+/-1.5 follicles), while it increased in October (3.9+/-1.2 follicles; P<0.05). Mean progesterone concentrations were lowest in January (0.3+/-0.1 ng mL(-1)) when compared with March (3.5+/-1.8 ng mL(-1); P=0.063), June (4.4+/-1.4 ng mL(-1); P<0.05) and October (2.2+/-0.9 ng mL(-1); P<0.05). At Day 2 of PRID treatment, mean progesterone concentrations significantly increased in all mares. Except from January, mean LH concentrations decreased within one day after PRID insertion and remained at low levels during treatments in January and March. Total secretion of LH during PRID-treatment was significantly lower in January and March when compared with June and October. In the 5 of 7 mares that ovulated during PRID treatment a distinct increase of plasma LH concentrations after ovulation was detected. Administration of the progesterone releasing intravaginal device PRID combined with the PGF2alpha analogue cloprostenol was able to induce estrus and ovulation in mares at different times of the year. However, efficacy of the treatment was not satisfactory concerning effectiveness in relation to season and synchrony of intervals from removal of PRID to ovulation in mares.

Administration, Intravaginal↗

Active immunization against follistatin and its effect on FSH, follicle development and superovulation in heifers.

Ovaries of heifers were examined daily by transrectal ultrasonography for one interovulatory interval before initiation of immunizations (control cycle, n = 14), and again after the fifth immunization with a sham-vaccine (Freund's adjuvant only; n = 7) or a recombinant porcine follistatin-vaccine (1 mg per vaccination; n = 7) to study the effect of follistatin on follicle dynamics. After the fifth immunization, 4 heifers had a follistatin antibody titer of > or = 1:3200, while the remaining 3 heifers had a titer of only 1:400. At wave emergence, the total number of follicles and the number of small follicles (3 to 5 mm) were higher (P < 0.05) in the follistatin group than in the control and sham groups. In addition, high-titer heifers had a greater (P < 0.05) number of follicles (total and small) per day than low-titer heifers. Plasma concentration of FSH remained unchanged after sham- or follistatin-immunization. Sham- and follistatin-vaccinated heifers were then given half the standard superovulatory dose of Folltropin (200 mg of FSH) 14 d after the sixth immunization. More ovulations were detected in follistatin- (10.9 +/- 2.4) than sham- (5.0 +/- 0.8) vaccinated heifers (P < 0.05). Moreover, heifers with a high titer had more ovulations (P < 0.02) than heifers with a low titer (15.0 +/- 2.5 vs 5.3 +/- 1.2). The number of ova-embryos classified as fertilized:unfertilized and transferable:discarded, and quality of the embryos were similar between sham and follistatin groups. By 80 d after the last immunization, when antibody titers were undetectable in the follistatin group, there was no difference in superovulatory response between sham (6.7 +/- 1.6) and follistatin (7.6 +/- 1.6) groups. In summary, follistatin immunization was associated with an increase in the number of small follicles at the time of wave emergence and a greater response to superovulatory treatment. The results suggest that effects of follistatin on follicular dynamics were not mediated through changes in pituitary secretion of FSH.

Animals↗

A comparative evaluation of the safety and contraceptive effectiveness of 65 mg and 100 mg of 90-day norethindrone (NET) injectable microspheres: a multicenter study.

The first of a second generation of slow-release injectable contraceptives is the norethindrone (NET) microspheres with a 90-day duration of action. It was evaluated at 65-mg and 100-mg doses for safety and contraceptive effectiveness in two randomized, single-blind trials among 131 women: 94 women for 12 months and 37 women for 6 months. The 6-month trial included additional evaluations of ovarian function and serum NET values. In the 6-month trial, no indication of ovulation was detected in the 100-mg dose group, while 3 of the 19 women in the 65-mg group showed signs of ovulation (progesterone greater than 3 ng/ml). No pregnancies were reported in the 100-mg group and one pregnancy in the 65-mg group resulted in a life-table pregnancy rate for that dose of 2.6 per 100 woman-years (95% confidence interval, 0 to 7.5). Days of vaginal bleeding were analyzed for 30 days before treatment and in 90-day reference periods after treatment. The mean number of vaginal bleeding and spotting days increased initially after the first injection in both dose groups, but decreased to below baseline in both dose groups after 6 months. The two doses appear comparable in clinical safety, side effects, vaginal bleeding patterns, and laboratory measures. With the preliminary estimate of efficacy, the 65-mg dose would be the minimally effective dose for the NET 90-day injectable contraceptive.

Adult↗

Incidence of spontaneous ovulation and development of the corpus luteum in non-mated dromedary camels (Camelus dromedarius).

The occurrence of spontaneous ovulation in dromedaries was examined in two separate studies including 20 non-lactating, barren and 12 lactating dromedaries, respectively. Lactating camels were milked twice a day with an automatic bucket milking machine. Ovarian activity was monitored by repeated ultrasonography. Blood samples for progesterone were collected daily or two to three times a week. To compare CL development after spontaneous and induced ovulations, ovulation was induced by a GnRH analogue in eight lactating dromedaries. Spontaneous ovulation was observed in one non-lactating camel (1 of 20 camels, 5%; 1 of 70 follicular waves, 1.4%), whereas, spontaneous ovulation was detected more frequently in lactating dromedaries (5 of 12 camels, 41.7%; 13 of 91 follicular waves, 14.3%). In one lactating camel, spontaneous ovulation occurred repeatedly for nine times. There was a significant effect of type of ovulation (spontaneous versus induced, P < 0.05) and day (P < 0.001) on serum progesterone concentration. Mean serum progesterone levels and total progesterone production (AUC) were higher after induced ovulation. Luteal diameter and serum progesterone concentration were positively correlated (r = 0.71, P < 0.001), but there was a significant difference between morphological and functional development of the CL. In dromedaries, morphological development starts earlier, morphological regression starts later and last longer than functional development and regression of the CL. Compared to induced ovulation, functional development of the CL after spontaneous ovulation might be altered but the morphological development is not affected.

Animals↗

Influence of premature induction of a luteinizing hormone surge with gonadotropin-releasing hormone on ovulation, luteal function, and fertility in cattle.

We tested the hypothesis that luteal function and fertility would be reduced in cattle induced to ovulate prematurely compared with those ovulating spontaneously. Estrus was synchronized in 56 beef cows (24 that were nonlactating and 32 that were nursing calves). At 6.4 +/- 0.1 d after estrus, all follicles > or = 5 mm were aspirated (day of aspiration = d 0) with a 17-gauge needle using the ultrasound-guided transvaginal approach. On d 1.5 and 2, cows were administered 2 luteolytic doses of PGF2alpha. Ovarian structures were monitored by transrectal ultrasonography from d -2 to 12, or ovulation. Emergence of a new follicular wave occurred on d 1.7 +/- 0.1. When the largest follicle of the newly emerged wave was 10 mm in diameter (d 4.8 +/- 0.1), cows were assigned on an alternating basis to receive 100 microg of GnRH (GnRH-10; n = 29) to induce ovulation or, upon detection of spontaneous estrus, to the spontaneous (SPON) treatment (n = 24). Cows were bred by AI at 12 h after GnRH (GnRH-10) or 12 h after the onset of estrus (SPON) as detected using an electronic surveillance system. Blood samples were collected every other day beginning 2 d after ovulation until pregnancy diagnosis 30 d after AI. Ovulation and AI occurred in 29/29 cows in the GnRH-10 and in 24/24 cows in the SPON treatment. Ovulation occurred later (P < 0.05) in the SPON (d 7.7 +/- 0.1) than GnRH-10 (d 6.8 +/- 0.1) treatment. Double ovulations were detected in 47% of cows, resulting in 1.5 +/- 0.1 ovulations per cow. Diameters of the ovulatory and the second ovulatory (in cows with 2 ovulations) follicles were greater (P < 0.05) in the SPON (12.0 +/- 0.3 mm and 10.5 +/- 0.4 mm, respectively) than in the GnRH-10 (10.7 +/- 0.1 mm and 9.2 +/- 0.3 mm) treatment. Cross-sectional areas of luteal tissue and plasma concentrations of progesterone during the midluteal phase were greater (P < 0.05) in the SPON (3.62 +/- 0.2 cm2 and 6.4 +/- 0.3 ng/mL) than in the GnRH-10 (3.0 +/- 0.2 cm2 and 5.4 +/- 0.2 ng/mL) treatment. The conception rate to AI in the SPON (100%) treatment was greater (P < 0.05) than in the GnRH-10 (76%) treatment. The animal model used in this study resulted in unusually high conception rates and double ovulations. In conclusion, premature induction of the LH surge reduced the diameter of ovulatory follicle(s), the luteal function, and the conception rate to AI.

Animals↗

Fine mapping a quantitative trait locus affecting ovulation rate in swine on chromosome 8.

Ovulation rate is an integral component of litter size in swine, but is difficult to directly select for in commercial swine production. Because a QTL has been detected for ovulation rate at the terminal end of chromosome 8p, genetic markers for this QTL would enable direct selection for ovulation rate in both males and females. Eleven genes from human chromosome 4p16-p15, as well as one physiological candidate gene, were genetically mapped in the pig. Large insert swine genomic libraries were screened, clones were isolated and then screened for microsatellite repeats, and informative microsatellite markers were developed for seven genes (GNRHR, IDUA, MAN2B2, MSX1, PDE6B, PPP2R2C, and RGS12). Three genes (LRPAP1, GPRK2L, and FLJ20425) were mapped using genotyping assays developed from single nucleotide polymorphisms. Two genes were assigned since they were present in clones that contained mapped markers (HGFAC and HMX1). The resulting linkage map of pig chromosome 8 contains markers associated with 14 genes in the first 27 cM. One inversion spanning at least 3 Mb in the human genome was detected; all other differences could be explained by resolution of mapping techniques used. Fourteen of the most informative microsatellite markers in the first 27 cM of the map were genotyped across the entire MARC swine resource population, increasing the number of markers typed from 2 to 14 and more than doubling the number ofgenotyped animals with ovulation rate data (295 to 600). Results from the revised data set for the QTL analysis, assuming breed specific QTL alleles, indicated that the most likely position of the QTL resided at 4.85 cM on the new linkage map (F1,592 = 20.5150, genome-wide probability less than 0.015). The updated estimate of the effect of an allele substitution was -1.65 ova for the Meishan allele. The F-ratio peak was closest to markers for MAN2B2 (4.80 cM) and was flanked on the other side by markers for PPP2R2C. Two positional candidate genes included in this study are MAN2B2 and RGS12. These results validate the presence of a QTL affecting ovulation rate on chromosome 8 and facilitate selection of positional candidate genes to be evaluated.

Animals↗

Cyclic changes of cervical mucus enzymes related to the time of ovulation. I. Alkaline phosphatase.

The concentration of alkaline phosphatase in cervical mucus was serially determined during a menstrual cycle in five normal ovulatory women and correlated with the time of ovulation as monitored by the basal body temperature and radioimmunoassay of serum lutenizing hormone (LH), progesterone, and estradiol. The activity of alkaline phosphatase decreased significantly at midcycle just prior to the LH surge and began to rise after ovulation. Self-detection of cervical mucus alkaline phosphatase may provide a practical method of ovulation prediction.

Adult↗

Floral genes expressed in tomato hypocotyl explants in liquid culture.

This paper confirms, at molecular level, previous data showing that small explants of many plants do form a floral meristem and express specific floral genes after only few days in culture. After 15-20 days of culture, small tomato hypocotyl explants develop differentiated structures often resembling primitive ancestral reproductive organs. Other specific reproductive functions such as chromosomal segregation (somatic meiosis) were also present and demonstrated by means of a cytological and histological analysis. By reverse transcriptase-PCR and in situ hybridization it was found that these structures are indeed able to express flower-specific genes. The TM8 gene, a tomato gene that is expressed very early during floral development, is detectable on the proliferating hypocotyl explants during the first week of culture. The MON9612 gene, which in vivo is expressed only by tomato pistils and ovules, is detectable on the ovulelike structures developed after 20 days of culture. The construction of transgenic tomato plants expressing the GUS gene under the control of the MON9612 promoter allowed us to follow the induction and the expression of this gene during explant proliferation and development of the flowerlike structures. These data confirm the hypothesis that a floral reprogramming can be induced in plant explants as a consequence of wounding and growth factors action. It appears to be an effort to survive stress by means of an unscheduled reproductive program.

Culture Techniques↗

Administration of gonadotropin-releasing hormone during metoestrus in cattle: influence on luteal function and cycle length.

Gonadotropin-releasing hormone (GnRH) has been used to warrant the success of artificial insemination by accurately timing occurrence of ovulation. In practical conditions, GnRH may be administered too late, after ovulation, with an eventual reduction in pregnancy rate. The aim of this study was to investigate whether GnRH administration after ovulation would have a negative effect on luteal function. Three cows and six heifers of the Finnish Ayrshire breed were used. Oestruses were synchronised. After detection of ovulation, one of the following treatments was implemented: gonadorelin (250 microg, i.m.) at either 0-24h (T1) or 24-48h (T2) post-ovulation or control (no gonadorelin, C). Every animal was assigned once to each of these three manipulations. Ultrasonography was performed on days 1, 4 or 5, 7 or 8, 11 or 12, 14 or 15 post-ovulation and daily from the beginning of the next oestrous signs until ovulation (day 0=day of ovulation). Blood samples for progesterone (P(4)) determinations were collected daily from day 1 after the occurrence of ovulation until recording of the next oestrus. Administration of GnRH during metoestrus did not induce ovulation of either large or small follicles and, thus, no accessory corpora lutea (CL) were formed. In T1, on day 14 or 15, the diameter of CL was 1.3+/-0.3mm smaller than in C (P<0.01), but no differences were found either on days 11 or 12 or on the same days of the T2 and C treatments. No significant differences in levels or profiles of P(4) curves were found between GnRH treatments and control. Neither had the treatments any effects on the length of the oestrous cycle. In conclusion, GnRH treatment during metoestrus does not seem to alter subsequent luteal function and, thus, this does not explain previous reports of reduced fertility post-treatment.

Animals↗

A randomized controlled trial evaluating metformin pre-treatment and co-administration in non-obese insulin-resistant women with polycystic ovary syndrome treated with controlled ovarian stimulation plus timed intercourse or intrauterine insemination.

BACKGROUND: There are few data in the literature regarding the utility of metfomin before and during gonadotrophin administration in women with polycystic ovary syndrome (PCOS). The aim of the present study was to assess the effect of the pre-treatment and co-administration of metformin in infertile PCOS women treated with controlled ovarian stimulation (COS) followed by timed intercourse (TI) or intrauterine insemination (IUI). METHODS: Seventy insulin-resistant primary infertile women with PCOS were randomized to receive metformin cloridrate (850 mg twice daily; group A) or placebo tablets (two tablets daily; group B) for 3 months. Three trials of COS using highly purified urinary FSH (hpFSH) plus TI/IUI were performed. Number of ampoules of gonadotrophin used, duration of the ovarian stimulation, cycle cancellation, ovulation, pregnancy, abortion, live birth, mono-ovulatory cycles, multiple pregnancies and ovarian hyperstimulation syndrome (OHSS) rates were assessed. RESULTS: No difference between groups was detected in ovulation, cycle cancellation, pregnancy, abortion, live birth, multiple pregnancies and OHSS rates. The mono-ovulatory cycle rates were significantly (P = 0.002) more frequent in group A than in group B, whereas the days of stimulation for non-cancelled cycles and the number of vials of gonadotrophins used were significantly (P < 0.001) higher in group A than in group B. CONCLUSION: In insulin-resistant women with PCOS, metformin pre-treatment and co-administration with hpFSH increases the mono-ovulatory cycles.

Adult↗