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Behavioral indicators of ovarian phase in white-faced capuchins (Cebus capucinus).

In many primate species, conspicuous behavioral and/or morphological changes are indicators of the fertile phase of the female cycle. However, several primate species, such as the white-faced capuchin, lack these cues. This is referred to as "concealed ovulation," and is argued to be a reproductive strategy that confuses paternity and lowers the risk of infanticide. We studied 10 adult female white-faced capuchins in Santa Rosa National Park, Costa Rica, from January to June 2002. We determined their ovarian cycling patterns by analyzing fecal ovarian hormones, and compared simultaneously collected behavioral data to determine which, if any, cues females use to signal their fertile phases. We found that four females cycled during the study period but ceased to cycle without becoming pregnant. We considered several explanations for the lack of conception during our study, including reproductive seasonality. We found that female C. capucinus showed only small increases in rates of affiliative/proceptive behaviors directed toward adult males during their periovulatory phases. The best indicator of cycle phase was a significant increase in male affiliative behaviors (e.g., following and grooming bouts) and sexual behaviors (e.g., copulations and courtship displays) directed toward females during the periovulatory phase compared to the nonovulatory phase. Our finding that females exhibit little proceptive behavior, but that copulations and male courtship are nonetheless concentrated in periovulatory phases suggests that even though females do not provide behavioral and morphological cues to ovulation, males are still able to detect it. Infanticide occurs with some frequency in these monkeys, and there is evidence for postconceptive mating as a female strategy to lower risk of infanticide via paternity confusion. However, despite this occurrence of nonconceptive mating and the absence of female cues to ovulation, truly concealed ovulation does not appear to be characteristic of this study population of white-faced capuchins.

Animals↗

[Menstrual cycle lengths and the estimated time of ovulation in the cynomolgus monkey (Macaca fascicularis)].

Establishing of an efficient system for the judgement of an optimal mating time is an important prerequisite for the successful development of a large scale breeding of macaques as laboratory animals. In general, an optimal mating time has been judged on the basis of individual records of menstruation. The length of the previous menstrual cycle has usually been used for the prediction of the next cycle length. In the present study, the lengths of 240 menstrual cycles of 60 cynomolgus monkeys of our breeding colony were statistically analysed. The mean and the standard deviation of those cycle-lengths were 29.4 and 4.3 days, respectively. Correlationship between the length of a menstrual cycle and the length of its previous cycle was statistically significant (p less than 0.05). However, practically the correlation was not meaningful because the correlation coefficient (r) was very small even if the length of the second one of two successive cycles (r = 0.26) or the average length of three successive cycles (r = 0.36) is selected as a reference length. In other words, a cycle length can not be predicted on the basis of the length of the previous menstrual cycle. However, a marked increase of serum follicle stimulating hormone (FSH) concentration was observed from 8 to 15 days after menstruation. In 60 per cent of the animals observed, FSH increased 10 to 11 days after menstruation. Ovulation was observed one or two days later than the day of FSH increase. Therefore, the optimal mating time is judged to be between 11 and 14 days after menstruation regardless of the menstrual cycle length.

Animals↗

A novel basal lamina matrix of the stratified epithelium of the ovarian follicle.

Basal laminas are important sheets of specialized extracellular matrix that underlie and surround groups of cells, such as epithelia or endothelia, enabling the cells to orientate their basal/apical polarity and creating a microenvironment for them. Basal laminas can also individually encapsulate whole cells, such as muscle cells, thereby forming a microenvironment but not polarizing the enclosed cells. Other mesenchymal or stromal cells exist with no basal lamina. In the course of studying the bovine follicular basal lamina which underlies the multilayered epithelium of the ovarian follicle, we identified a developmentally regulated novel extracellular matrix (which we call focimatrix for focal intra-epithelial matrix). Focimatrix is composed of basal lamina-like material deposited as plaques or aggregates between the multilayers of the epithelial granulosa cells. The focimatrix does not encapsulate individual or groups of cells and therefore does not form a microenvironment for them. Focimatrix contains collagen type IV subunits alpha1 and alpha2 (but not alpha3-alpha6), and laminin chains alpha1, beta2 and gamma1 (but not alpha2 or beta1), and nidogen-1 and perlecan (but not versican). The amount of focimatrix increases with increasing follicular size, and its appearance precedes the expression by granulosa cells of the enzymes for steroid hormone synthesis, cholesterol side-chain cleavage cytochrome P450 (SCC) and 3beta-hydroxysteroid dehydrogenase (3beta-HSD), in the days preceding ovulation. The expression in granulosa cells of two components examined, nidogen-1 and perlecan, also increases substantially when follicles enlarge to a sufficient size capable of ovulating. Following ovulation the follicular basal lamina is degraded, and presumably focimatrix is too since it is not detected in corpora lutea that develop from the ovulating follicles. During this development the granulosa cells undergo an epithelial-mesenchymal transition (EMT) into luteal cells following ovulation, and substantially increase their expression of steroidogenic enzymes in the process. During EMT epithelial cells lose polarity. Since focimatrix exists on more than one side of the granulosa cells, we propose that it disrupts the polarity induced by the follicular basal lamina in the lead up to ovulation. Hence focimatrix maybe a key part of the follicular/luteal EMT.

Animals↗

Urinary luteinizing hormone testing and prediction of ovulation in spontaneous, clomiphene citrate and human menopausal gonadotropin-stimulated cycles. A clinical evaluation.

A urinary luteinizing hormone test was utilized to predict ovulation in 99 spontaneous, 122 clomiphene citrate, and 82 human menopausal gonadotropin stimulated cycles. Tests were performed in early morning and evening specimens and follicular development was monitored by daily ultrasonography. A positive detection rate of 98, 97, and 94%, respectively, was obtained. Evidence of luteinized unruptured follicles was seen more frequently in stimulated cycles, concurring with negative test results. In 2 spontaneous, 1 clomiphene citrate and 5 hMG induced cycles two distinct LH surges were detected concomitant with a pattern of follicular atresia and subsequent new follicular development. Most ovulations occurred between 16 and 28 h after LH detection, significantly earlier in spontaneous than in clomiphene citrate stimulated cycles (p less than 0.02), whereas pre-ovulatory follicles were larger in the clomiphene citrate group (p less than 0.001). The mean duration of the follicular and luteal phases, as calculated from the LH peak, was substantially shorter in the hMG cycles than in the other two groups (p less than 0.001).

Adult↗

The effects of administration of gonadotrophins and estrogens on prenatal survival in gilts.

In gilts ovulation occurs over a 4 to 8-hour period, with 70% of the ova being shed over a relatively short span of time. These oocytes supposedly give rise to more developed embryos at Days 10 to 12 which advance the uterine environment and reduce survival rates of less developed embryos because of an asynchronous environment. The aim of this experiment was to reduce embryo mortality by influencing the duration and pattern of ovulation. Crossbred gilts (n=98) were bred at their first observed estrus after being exposed to boars at 200 days of age. Estrus detection was carried out daily at 0000, 0800 and 1600 hours. All gilts were artificially inseminated with fresh semen, with a minimum of 2.7 billion spermatozoa, at both 16 and 32 hours after detection of estrus. Gilts were randomly assigned to one of the following treatments at detection of estrus: 1) 500 IU (2 ml) chorionic gonadotrophin (hCG) injected intravenously at the onset of estrus (n=22); 2) 16 microg (4 ml) gonadotrophin releasing hormone (GnRH) injected intravenously at the onset of estrus (n=25); 3) 11.5 microg estrogen added to the semen at the time of AI (n=25); 4) control, untreated gilts (n=26). All gilts were slaughtered at Day 30 of gestation (Day 0=day of detected estrus). The mean (+/-SEM) number of ovulations in pregnant gilts per treatment was 13.0+/-0.52, 12.6+/-0.51, 13.6+/-0.54 and 13.3+/-0.52, while the mean (+/-SEM) number of normal embryos per treatment was 10.3+/-0.67, 10.5+/-0.66, 10.3+/-0.69 and 10.5+/-0.67 for hCG, GnRH, estrogen and control groups, respectively, for an embryonic survival rate of 80+/-4.2%, 83+/-4.1%, 74+/-4.3% and 79+/-4.2% in pregnant gilts. If nonpregnant gilts are included, the embryonic survival rate for treatments 1 to 4 was 76+/-7.0%, 73+/-6.5%, 60+/-6.5%, and 64+/-6.4%, respectively. There was no significant difference between treatments for any of these variables. There was no evidence that administration of hCG, or GnRH at the onset of estrus, or the addition of estrogen to semen improved embryonic survival in gilts by Day 30 in this experiment.

Journal Article↗

Association of unexplained infertility with gonadotropin and ovarian antibodies.

PROBLEM: To determine the prevalence and characteristics of gonadotropin autoantibodies (GAB) associated with unexplained infertility, and to assess the relationship between ovarian autoantibodies (OVAB) and GAB. METHOD OF STUDY: Ovarian antibodies and GABs in sera of patients with unexplained infertility (n = 53) and a comparison (population) group from a blood bank (n = 40) were detected by immunoassay. Patients with unexplained infertility had either no prior gonadotropin treatment (n = 15) or two or more gonadotropin cycles to induce ovulation (n = 38). RESULTS: The GABs were detected in 67% of women with treatment, 27% of women without treatment and 8% of women in the population. The GABs recognized follicle stimulating hormone (FSH) and luteinizing hormone (LH) and their alpha and beta subunits and to a lesser extent thyroid stimulating hormone (TSH) and prolactin. There was no significant difference in OVAB prevalence between gonadotropin treated or untreated women while GABs were significantly more frequent in gonadotropin treated women (P < or = 0.01). CONCLUSION: Gonadotropin antibodies may represent a separate marker of ovarian autoimmunity in unexplained infertility as they are correlated with OVAB and are present in patients not treated with gonadotropin. However, as a higher frequency of GAB is associated with gonadotropin treatment, patients with ovarian autoimmunity may tend to have an immune response to gonadotropins in addition to an endocrine response.

Adolescent↗

Identification and opsonic activity of immunoglobulins recognizing Streptococcus zooepidemicus antigens in uterine fluids of mares.

A direct ELISA was used to measure immunoglobulin (Ig) isotypes G, Gt, A, and M recognizing Streptococcus zooepidemicus epitopes in uterine lavage fluids collected during the early post ovulatory period. A S. zooepidemicus isolate, used as the plate antigen in this assay, was inoculated into the uteri of 8 mares (3 resistant and 5 susceptible to endometritis) at oestrus prior to ovulation during Oestrous Cycles 1, 3 and 5. Resistant mares aged 2-5 years were nulliparous, with clinically normal reproductive tracts as determined by physical examination, bacteriological culture of the uterus, and endometrial biopsy. Susceptible, pluriparous mares, aged 16-24 years, had histories of endometritis and infertility and had chronic endometrial inflammatory changes detected by uterine biopsy. On Post ovulation Day 3 during Oestrous Cycle 1, on the day of ovulation during Oestrous Cycle 3 and on Post Ovulation Day 5 during Oestrous Cycle 5, uterine swab specimens were obtained for bacteriological culture and the uterus was irrigated with 50 ml sterile physiological saline solution containing tracer amounts of 125I-labelled human serum albumin (HSA). The titre of each Ig isotype was determined and the amount in uterine effluents was calculated from the dilution of the 125I-HSA. Total IgG concentration in uterine effluents was measured by radial immunodiffusion. Uterine fluids, either unheated or heated 30 min at 56 degrees C, were used to opsonize the S. zooepidemicus isolate in an assay of luminol-enhanced neutrophil chemiluminescence (CL). Mean peak CL generated was compared with CL generated by a control serum. Streptococcus zooepidemicus was isolated from 10 of 15 (67%) uterine specimens from susceptible mares.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Fertility of fully breast-feeding women in the early postpartum period.

OBJECTIVE: To examine bleeding between 6 and 8 weeks postpartum in fully breast-feeding women and its association with fertility as assessed by hormone analysis. METHODS: Seventy-two fully breast-feeding women were followed prospectively from 42 days postpartum. Vaginal bleeding was recorded daily. Women who experienced bleeding were compared with women who did not with respect to time of ovulation and time of first menses. RESULTS: Nearly half of the women experienced some vaginal bleeding or spotting between 6 and 8 weeks postpartum. These women eventually menstruated and ovulated earlier than the women who did not bleed, but the differences were not significant. The study had 34% and 45% power to detect a 20% difference in the proportion menstruating and ovulating, respectively, at 6 months postpartum, and 10% and 16% power to detect the same differences at 1 year. Seven women experienced ovarian follicular development before day 56, but neither bleeding nor follicular development was associated with ovulation in any woman in the first 8 weeks postpartum. CONCLUSIONS: It is unlikely that vaginal bleeding in fully breast-feeding women in the first 8 weeks postpartum represents a return to fertility.

Female↗

GnRH antagonist inhibition of luteinizing hormone secretion and ovulation in the pig.

A GnRH antagonist, Ac-D-p-Cl-Phe1,2, D-Trp3, D-Arg6, D-Ala10 GnRHb (Organon), was utilized to determine the effective dosage and duration to inhibit LH secretion in the pig. In a preliminary trial, barrows received either 10, 50, or 250 micrograms/kg BW of the GnRH antagonist. Secretion of LH was inhibited within 30 min for a duration of 12 h with the 100 micrograms/kg dose but persisted for greater than 48 h with the 250 micrograms/kg treatment. A second study determined effectiveness of the antagonist for inhibiting ovulation in cyclic gilts. At first detection of standing estrus, cyclic gilts were treated with either saline (control), 100, or 200 micrograms/kg BW of the GnRH antagonist (GnRH1). A second group of GnRH antagonist gilts received 200 micrograms/kg BW of the GnRH antagonist approximately 8 h prior to standing estrus (GnRH2). The GnRH1-treatment failed to inhibit or delay ovulation. Ovulation was inhibited and estrous cycles lengthened in GnRH2-treated gilts. These preliminary results suggest that ovulation in the gilt can be inhibited if the GnRH antagonist is administered prior to the LH surge.

Animals↗

Luteinizing hormone and ovulation timing in a therapeutic donor insemination program using frozen semen.

A series of 110 therapeutic donor insemination cycles was analyzed to determine the impact on fecundity when a urinary luteinizing hormone detection kit was used to plan inseminations. To prevent the transmission of human immunodeficiency virus, frozen semen, thawed after a 90-day quarantine, was used. The minimum standard for insemination with cryopreserved semen was a total of 24 x 10(6) motile sperm per milliliter after thawing. Fecundity was 0.12 when insemination timing was based on cervical mucus evaluation and basal body temperature charts and 0.13 when a urinary luteinizing hormone kit was also used to predict ovulation. Life table analysis with the log rank test showed no statistically significant difference in the number of cycles required to achieve conception between the group of patients using conventional methods of ovulation timing and the group of patients using the urinary luteinizing hormone kit. Urinary luteinizing hormone testing offers no advantage over conventional methods, such as cervical mucus examination and evaluation of basal body temperature, when ovulation is being timed for insemination with frozen donor semen.

Acquired Immunodeficiency Syndrome↗

Corticosteroids do not influence ovarian responsiveness to gonadotropins in patients with premature ovarian failure: a randomized, placebo-controlled trial.

OBJECTIVE: To determine the effect of corticosteroids on ovarian responsiveness to exogenous gonadotropins in patients with idiopathic premature ovarian failure (POF). DESIGN: Placebo-controlled, randomized, double-blind, multicenter study. SETTING: Two tertiary care academic centers for reproductive endocrinology and fertility and two general teaching hospitals. PATIENT(S): One hundred patients with idiopathic POF intended to enter the study. The study was discontinued after 36 patients failed to ovulate. INTERVENTION(S): Endocrine and immune parameters were tested on days 1 and 15. On day 1, subjects were randomized to receive either 9 mg of dexamethasone daily or placebo. From day 5 onward, 300 IU of hMG daily was added for 10 days in both groups. The dosage of dexamethasone was decreased stepwise in the second week and discontinued after day 15. Patients were monitored by transvaginal ultrasonography and by determining serum E2 levels. MAIN OUTCOME MEASURE(S): Ovulation rate. Fifty patients would have to be included in each study group to detect a statistically significant difference of 20% in the ovulation rate between the two groups with alpha = 0.05 and beta = 0.1 (one-tailed test). RESULT(S): No ovulation was recorded in the first 36 patients. Interim analysis showed that the 95% confidence intervals of an ovulation rate of 0 were 0-17% for the dexamethasone arm (n = 19) and 0-19% for the placebo arm (n = 17). Because the preset objective (a difference of 20%) would never be reached, the study was discontinued. CONCLUSION(S): Corticosteroids do not influence ovarian responsiveness to gonadotropins in patients with idiopathic POF.

Adrenal Cortex Hormones↗

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.

Animals↗

The effect of the intramammary infusion of Escherichia coli endotoxin on ovulation in lactating dairy cows.

The purpose of this experiment was to determine if intramammary inflammation during the periovulatory period affects the occurrence of ovulation in lactating dairy cows. Ten lactating, cyclic, Holstein dairy cows received 2 injections of prostaglandin F2alpha at eleven-day intervals, to synchronize luteolysis. The day of the second injection was designated as day 0. Ovulation was anticipated to occur 3-5 days later (on days 3-5). Beginning at the morning milking on day 1, cows received intramammary infusions of either Escherichia coli endotoxin (10 microg; n=5) or infusion vehicle (pyrogen free Hank's balanced salt solution; n=5) into 2 quarters immediately after milking. The same quarters were infused after each milking through day 4. Venous blood samples were collected daily from day -1 through 13 for determination of progesterone to monitor luteolysis and formation of a new corpus luteum. Blood samples were also collected at 4-h intervals (days -1 to 2), then at 2-h intervals (days 2 to 5) to measure concentrations of luteinizing hormone. Ovaries were examined ultrasonographically on days -1 through 5 and on day 12 to monitor follicular growth and formation of the corpus luteum. Collectively, these observations were used to determine if and when ovulation occurred. Intramammary infusion of E. coli endotoxin induced an immediate increase in the concentration of somatic cells in milk from treated quarters. However, this treatment had no effect on the occurrence or timing of ovulation. Based on ultrasonography and concentrations of progesterone, four of five cows in each treatment group appeared to have ovulated. Preovulatory surges of LH were detected within the intensive bleeding periods for three cows in each treatment group. The magnitude of the LH surge was reduced in cows receiving endotoxin.

Animals↗

Early-weaned sows: altrenogest therapy, estrus, ovulation, and reproductive performance.

Early weaning is a technique used to increase swine health status, and may cause consequences in reproductive performance of sows. An experiment was performed to evaluate these effects in a herd of sows, with weaning at 9 or 10 days post-farrowing, located in west of Minas Gerais, Brazil. Large-White sows (n=102), with three or four previous parturitions were randomly allocated to three treatment groups: T1: artificial insemination (AI) at first post-weaning estrus of the sows; T2: AI at second post-weaning estrus, T3: AI at first estrus, after an administration of a daily individual dose of 20 mg of altrenogest from 5 to 8 days post-weaning. The duration of the first post-weaning estrus did not differ among treatment groups; however, the second estrus of the T2 group was of shorter duration relative to the other treatment groups (P< or =0.035). Ovulation occurred earlier at the second estrus of the T2 group, compared with the T1 and T3 groups (P< or =0.027), being similar to that at the first estrus of T2 group (P=0.177). The relationship of the timing between ovulation and estrus was similar among treatment groups (P> or =0.221). There was no difference in farrowing rate among treatment groups (P> or =0.313). The T2 group produced a mean of 2.5 more piglets per litter (P=0.002). In conclusion, the use of altrenogest did not increase the reproductive performance of early-weaned sows.

Animals↗

Rapid detection of the preovulatory luteinizing hormone peak in the blue fox (Alopex lagopus) by an enzyme-linked immunosorbent assay.

An enzyme-linked immunosorbent assay (ELISA) developed for the rapid measurement of luteinizing hormone (LH) in blood of many mammalian species has been validated in the fox. For this purpose, daily plasma samples collected from six vixens during the periovulatory period were assayed for LH using ELISA. The results were compared with those obtained by radioimmunoassay (RIA), and related to parameters used to determine the time of ovulation (onset of oestrus, vaginal electrical resistance peak) and to ovarian findings. This ELISA provided a highly sensitive measure of LH (125 pg ml-1) from a 10 microliters plasma sample within 3 h. Plasma concentrations of LH measured by ELISA and RIA were significantly correlated (r = 0.911). In the blue fox, ovulation occurs once a year and coincides with the start of oestrus. The LH peak preceded the start of oestrus by 1.5 days (1-3) and preceded the vaginal electrical resistance peak by 2.2 days (1-4). The rapid and precise detection of the preovulatory LH peak by this assay provides a convenient method to determine the time of ovulation and therefore the optimum time for artificial insemination or oocyte collection for fertilization in vitro.

Animals↗

Sperm distribution in the genital tract of the bitch following artificial insemination in relation to the time of ovulation.

In the present study, sperm distribution in the genital tract of the bitch following artificial insemination (AI) in relation to the time of ovulation was investigated by histology, scanning electron microscopy (SEM) and flushing. Ten bitches were inseminated intravaginally with 500 x 10(6) spermatozoa: three dogs before ovulation, four dogs during ovulation and three dogs after ovulation. Ovariohysterectomy was performed 24 h after AI. Half of the genital tract was divided into nine segments (cervix, corpus uteri, caudal, middle and cranial uterine horn (UTH), utero-tubal junction (UTJ), isthmus, ampulla and infundibulum), which were processed for histology and SEM. The contralateral UTH and uterine tube (UT) were flushed, and several sperm characteristics were assessed. Histology revealed that the spermatozoa were mainly located in the uterine glands and at the UTJ, while very few spermatozoa were detected in the UT. Insemination during ovulation resulted in higher percentages of glands with spermatozoa in the different parts of the uterus (P < 0.05). Evaluation by SEM showed higher numbers of spermatozoa in several parts of the uterus for bitches inseminated during ovulation (P < 0.05). The mean number of spermatozoa flushed from the UTH and the UT was low. No significant differences in the evaluated sperm quality parameters were found between the flushings of the UTH and the UT. In conclusion, based on our findings, the uterine glands and the UTJ might act as sperm reservoirs in the bitch and sperm transport in the genital tract is affected by the time of AI in relation to ovulation.

Animals↗

[Current determination of ovulation time. 1. Rapid LH determination in serum and urine].

Short-term recording of LH rise is a possible approach to ovulation forecast. Hormone values in serum and four-hour urine can be obtained in three and half hours by modification of a commercial radio-immunological LH test. The results were compared with data obtained from the HI Gonavis test. Observable LH rise can be detected between three and six hours earlier by LH serum measurement, as compared to collected urine. Suitable information may be obtained also from LH urine assay, a non-invasive method of ovulation prediction.

Female↗