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PubMed2005

JSAR Innovative Technology Award. Development of ovulation synchronization and fixed time artificial insemination in dairy cows.

Recently, reproductive management has become more difficult as a result of increased herd size. Problems with missing estrous signs and decrease in conception rate by artificial insemination (AI) performed at wrong timing have caused low AI conception rates. In 1995, ovulation synchronization and fixed-time AI (Ovsynch/TAI) was developed in the USA as a new reproductive technology, which was accepted as an useful reproductive management tool in many countries. However, no information on the use of Ovsynch/TAI was available in Japan. It was, therefore, warranted to show the ovulation rate and conception rate after Ovsynch/TAI using gonadotropin releasing hormone analogue (GnRH-A, fertirelin acetate) and prostaglandin F2alpha (PGF2alpha)-THAM, both were commercially available in this country. The conception rate after Ovsynch/TAI has been known to vary among different herds and individuals. Investigation and analysis of factors affecting the conception rate was also warranted to improve the conception rate. A series of experiments were carried out to establish Ovsynch/TAI using domestically produced GnRH-A and PGF2alpha and to study factors affecting conception rate after Ovsynch protocol. Ovsynch using 100 microg GnRH-A and 25 mg PGF2alpha were observed using ultrasonography. As a result, a high synchronization rate of ovulation at 16 to 20 h after the second GnRH injection was confirmed. The conception rate after Ovsynch/TAI was compared in 87 cows with the conception rate after AI at estrus induced by PGF2alpha (139 cows). Conception rate after Ovsynch/TAI was higher than the figure after AI at induced estrus (59.1% vs 20.9%, P<0.05). The dose of GnRH-A was also studied and a practical dose of GnRH-A was found to be 50 microg per cow. To clarify some factors affecting the conception rate after Ovsynch/TAI, 1,558 cows were investigated for the state of their ovaries, days after calving, parity, season, ovarian cyclicity postpartum and nutritional state at the day of Ovsynch. The overall conception rate after Ovsynch/TAI was 51.5%. Fifty-six cows (3.6%) showed estrus at 6 to 7 d after the first injection of GnRH-A. The conception rate after Ovsynch/TAI was low in cows that were 40 to 60 d postpartum, those in their 5th lactation or more, those bred in July to August, and those recovering ovarian cyclicity later than 56 d postpartum. The conception rate after Ovsynch/TAI was high in cows in which body condition score (BCS) was 3.75 at dry period and 3.0 at the day of Ovsynch. In conclusion, Ovsynch/TAI is an effective tool for the reproductive management of dairy cows. A steady and sufficient conception rate after Ovsynch/TAI could be expected by taking the factors affecting the conception rate into the consideration.

Animals↗
PubMed2001

Transfer of canine embryos at various developmental stages recovered by hysterectomy or surgical uterine flushing.

In dogs, embryo transfer (ET) techniques such as induciton of excessive ovulation and synchronization of estrus have not progressed well. Therefore, using embryos at various developmental stages, ET was investigated in dogs from a beagle colony in which the ovulation days were close, as estimated by the progesterone level. Embryos were, recovered 8-11 days after ovulation (4-9 days after mating) by excising the oviducts and uteri (excision method) in 16 animals and by surgical flushing of the uteri at laparotomy (surgical method) in 3 animals. In 24 dogs with -4 to +2 days of difference in the timing of ovulation between donor and recipient dogs, 1-10 embryos at the 8-cell to blastocyst stages were transferred per animal. The mean embryo recovery rate by the excision method (97.1%) was significantly higher than that by the surgical method (42.5%) (p<0.01). Twelve (57.1%) of 21 animals with -1 to +2 days difference in ovulation day became pregnant after the transfer of 8-cell to blastocyst stage embryos. Although 3 dogs with -4 to -2 days of difference of ovulation day underwent ET of morula or compacted morula, none of these dogs became pregnant. The mean ratio of the number of newborns to the number of transferred embryos was only 51.9%. The mean duration of the period between ovulation and delivery in the pregnant recipients was 65.8 days, which tended to be longer than that in natural mating. These results demonstrate that pregnancy can be induced by ET at the 8-cell to blastocyst stage in dogs with -1 to +2 days difference in ovulation day.

Animals↗
PubMed2002

The evaluation of infertility.

Infertility is a significant medical problem that affects many couples. Evaluation is the starting point for treatment of infertility as it may suggest specific causes and appropriate treatment modalities. Although the history and physical examination provide important information, specific diagnostic tests are required to evaluate infertility. Because the causes of infertility can be multifactorial, a systematic approach typically is used and involves testing for male factor, ovulatory factor, uterotubal factor, and peritoneal factor. Many of these diagnostic tests are laboratory based, including semen analysis, serum progesterone level, serum basal follicle-stimulating hormone level, and clomiphene citrate challenge, and can be done by the primary care physician. Moreover, by understanding the infertility evaluation, the primary care physician can serve as an important resource for advice about infertility. This article briefly reviews the diagnostic approach to infertility, with particular emphasis on important laboratory tests used for evaluation.

Adult↗
PubMed1979

The concentrations of urinary oestrone-3-glucuronide, LH and pregnanediol-3alpha-glucuronide as indices of ovarian function.

Methods based upon the principles of radioimmunoassay have been developed and evaluated for the measurement of oestrone-3-glucuronide, LH and pregnanediol-3alpha-glucuronide in samples of unextracted urine. The procedures have been applied to daily urine (early morning fraction and combined 24 hour collections) from 6 women throughout one complete menstrual cycle and to serial samples from an additional 14 women who only collected early morning specimens. The results showed that there were characteristic, well-defined changes in the concentration of all 3 metabolites in both samples of urine and from all subjects. In addition, there was a reasonable correlation between the concentration of all 3 compounds in samples of early morning urine and the corresponding rates of excretion per 24 hours. These findings suggest that the procedures may be of value for monitoring ovarian function over long periods of time, without the problems of stress and inconvenience to the patient. Furthermore, the ratio of values for oestrone-3-glucuronide to pregnanediol-3alpha-glucuronide may be used to indicate the start and finish of the fertile period.

Adult↗
PubMed1980

The effect of conception on the concentration of urinary oestrone-3-glucuronide, LH/hCG and pregnanediol-3 alpha-glucuronide.

The concentrations of oestrone-3-glucuronide, LH/hCG and pregnanediol-3 alpha-glucuronide have been measured in daily samples of early morning urine from 6 women throughout the normal ovarian cycle and from 5 subjects, who conceived during the period of study. In addition, the ratio of oestrone to pregnanediol glucuronide was calculate as an additional index of ovarian function. The results showed that the ratio during the follicular phase of the cycle was similar in both groups of women. Moreover, there was no significant difference between the groups in either the duration or intensity of the LH peak. Seven days later, however, the women who conceived excreted significantly higher amounts of oestrone-3-glucuronide (P less than 0.05) and 12 days after the LH peak the values for pregnanediol-3 alpha-glucuronide and LH/hCG were also elevated (P less than 0.05). A defined increment, in the concentration of hCH (3 times the mean basal level) occurred 11 to 15 days after the peak of LH and appeared 3 to 5 days after the increased excretion of the steroid glucuronides.

Adult↗
PubMed1977

A study of peroxidase levels in human cervical mucus as an index of ovulation.

Fourteen healthy women were screened for the following parameters throughout one complete menstrual cycle: levels of urinary oestrogens and pregnanediol; levels of plasma and urinary gonadotrophins; and pH, protein content and levels of peroxidase in the cervical mucus. It was found that concentrations of peroxidase in the cervical mucus were not a reliable index of ovulation.

Cervix Mucus↗
PubMed1982

Changes in the plasma concentrations of free and conjugated oestrogens in heifers after treatment to induce superovulation and the relationship with number of ovulations.

Oestradiol-17 beta and conjugated oestrone, oestradiol-17 beta and oestradiol-17 alpha were measured in peripheral plasma of heifers treated with PMSG/PGF-2 alpha to induce superovulation. Changes in the concentrations of each hormone were synchronous, the highest level being near oestrus. For a given number of ovulations the hormone with the highest concentration was total oestradiol-17 alpha, then came total oestrone, total oestradiol-17 beta and oestradiol-17 beta. For each oestrogen, the maximum preovulatory concentration measured was significantly correlated with the number of ovulations; the regression line for total oestradiol-17 alpha was twice as steep as that for oestradiol-17 beta. It is concluded that in animals treated to induce superovulation assay of total oestradiol-17 alpha gives a better induction of the number of follicles induced to ovulate than does the more conventional assay of oestradiol-17 beta.

Animals↗
PubMed1983

Prediction of bovine ovulation by a rapid radioimmunoassay for plasma LH.

A commonly used and specific plasma LH radioimmunoassay was modified to provide reliable results for cow LH concentrations in blood within 4-5 h. Blood samples were collected at 2- or 4-h intervals from at least 24 h before the expected oestrus until after its onset. Thereafter, samples were collected at intervals of 2 or 4 days until the next oestrus. For the assay, the antigen-antibody reaction took place during a 2-h incubation at 37 degrees C. The immune complex was precipitated by addition of rabbit anti-gamma-globulin and polyethylene glycol followed by an incubation at 22 degrees C for 30 min. The coefficient of variation between this assay and the standard assay was less than 15%. From a study of 25 cycles from 12 animals, we obtained intervals of congruent to 27.3 h between the beginning of the LH peak and ovulation and congruent to 17.5 h between the end of the peak and ovulation. This modified technique can be used to predict rapidly and precisely the time of ovulation in cattle.

Animals↗
PubMed1990

Urinary endocrine monitoring of the ovarian cycle and pregnancy in Goeldi's monkey (Callimico goeldii).

A non-invasive study of urinary hormones in 6 captive female Goeldi's monkeys provided accurate information on reproductive function. Conjugated oestrone accounted for 80-85% of the urinary oestrone and oestradiol measured. Radioimmunoassay measurements of conjugated oestrone provided a reliable indicator of cyclic ovarian function (mean cycle length: 24.1 +/- 0.9 days; n = 9) and pregnancy (gestation: 145, 155 days; n = 2). Measurements of urinary progesterone and pregnanediol glucuronide were only reliable as indicators of ovarian cyclicity. Elevations in urinary conjugated oestrone coincided with luteal-phase elevations of urinary progesterone and pregnanediol glucuronide. Urinary LH concentrations provided no indication of pituitary activity. However, the frequencies of female sexual solicitations of males were maximal when oestrone conjugate concentrations rose, suggesting a peri-ovulatory period. Ovulation was suppressed in 1 of 3 subordinate females housed in male-female-female trios.

Animals↗
PubMed1993

Determination of ovarian cycle in Goeldi's monkey (Callimico goeldii) via the measurement of steroids and peptides in plasma and urine.

Goeldi's monkey (Callimico goeldii) is an endangered species of New World primate. The present study provides the first description of the non-conception ovarian cycle in this species based on circulating reproductive steroid and peptide hormones. The data obtained were used to validate a non-invasive system for monitoring cyclicity based on urinary reproductive steroid metabolites. Nine sexually mature females were studied. In three females, matched blood and urine samples were collected once every 2-3 days for 90-120 days; in three other females, matched blood and urine samples were collected daily for 14-20 days for one peri-ovulatory period; and in the remaining three females, urine samples only were collected once every 1-3 days for 40-60 days. Plasma progesterone, oestrone-3-conjugates and bioactive LH were measured, in addition to urinary pregnanediol-3 alpha-glucuronide and oestrone-3-conjugates. The mean maximum concentration of plasma LH occurred 1-2 days before a significant rise in plasma progesterone, which was considered to occur 1 day after ovulation. On the basis of plasma progesterone titres, the duration of the ovarian cycle was estimated as 23.9 +/- 0.4 days (n = 9), and constituted a follicular phase of 10.7 +/- 0.3 days and a luteal phase of 13.5 +/- 0.3 days. Urinary pregnanediol-3 alpha-glucuronide demonstrated a high correlation with plasma progesterone (r = 0.8), and demonstrated a significant rise at the same time as plasma progesterone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1998

Ovarian follicular dynamics during anoestrus in ewes.

The aim of the present study was to document ovarian antral follicle dynamics throughout seasonal anoestrus in sheep. Daily transrectal ultrasonography was performed during four 17 day scanning periods from March to July in Western White-faced crossbred ewes. Blood samples were collected each day with ultrasonographic scanning for measurement of serum concentrations of FSH, oestradiol and progesterone. Blood samples were also taken every 15 min for 6 h, mid-way through each period of ultrasonographic examination, to determine the patterns of secretion of gonadotrophic hormones. Hormonal data were then related to observed changes in follicular populations and the patterns of antral ovarian follicle turnover. Ultrasonography showed that the ovaries of anoestrous ewes remained active and that the largest ovarian antral follicles grew to a periovulatory size (> or = 5 mm in diameter) at all stages of anoestrus. The total number of all ovarian follicles > or = 3 mm in diameter was lower during early anoestrus compared with at mid-anoestrus because of a significantly smaller number of small (3 mm) and medium (4 mm) ovarian follicles. The largest ovarian follicles (attaining > or = 5 mm in diameter before regression) exhibited a wave-like pattern of growth; an average of three waves of follicular development were recorded in sheep during each of the four 17 day scanning periods in anoestrus, with follicular waves emerging approximately every 5 days. This rhythmic pattern of follicular emergence was found to be associated with the occurrence of fluctuations in serum FSH concentrations. The growth rate of the largest follicles of the wave increased significantly from early to late anoestrus in sheep. In addition, ovarian follicles not growing beyond 3 mm in diameter showed organized patterns of growth and regression; their numbers tended to be lower (P = 0.09) at 3 days before and on the day of follicular wave emergence. Some ewes were seen to maintain synthesis of progesterone throughout anoestrus. This submaximal progesterone secretion tended to occur at irregular intervals and was not coupled with changes in concentrations or patterns of gonadotrophin release, ovulations or detectable morphological luteinization of ovarian antral follicles. It was concluded that the growth of ovarian antral follicles to an ovulatory size was maintained throughout anoestrus in ewes, with a transient shift in the number of small and medium-sized follicles during mid-anoestrus, and that the periodic emergence of waves of large follicles (> or = 5 mm in diameter) occurred in synchrony with an endogenous rhythm of FSH secretion.

Anestrus↗
PubMed2006

Fecal estradiol and progesterone metabolite levels in the three-toed sloth (Bradypus variegatus).

The present study was carried out to assess the possibility of measuring fecal steroid hormone metabolites as a noninvasive technique for monitoring reproductive function in the three-toed sloth, Bradypus variegatus. Levels of the estradiol (E2) and progesterone (P4) metabolites were measured by radioimmunoassay in fecal samples collected over 12 weeks from 4 captive female B. variegatus sloths. The validation of the radioimmunoassay for evaluation of fecal steroid metabolites was carried out by collecting 10 blood samples on the same day as defecation. There was a significant direct correlation between the plasma and fecal E2 and P4 levels (P < 0.05, Pearson's test), thereby validating this noninvasive technique for the study of the estrous cycle in these animals. Ovulation was detected in two sloths (SL03 and SL04) whose E2 levels reached 2237.43 and 6713.26 pg/g wet feces weight, respectively, for over four weeks, followed by an increase in P4 metabolites reaching 33.54 and 3242.68 ng/g wet feces weight, respectively. Interestingly, SL04, which presented higher levels of E2 and P4 metabolites, later gave birth to a healthy baby sloth. The results obtained indicate that this is a reliable technique for recording gonadal steroid secretion and thereby reproduction in sloths.

Animals↗
PubMed1976

On differences between the preovulatory luteinizing hormone surges of 4- and 5-day cyclic rats.

Preovulatory LH surges were studied in rats with ovarian cycles of 4 days (4d-rats) and 5 days (5d-rats). In 5d-rats the maximal peak height was about twice that observed in 4d-rats, whereas in 4d-rats peaks occurred about 1-5 h later than in 5d-rats and were much less consistently timed. From experiments in which LH releasing hormone (LH-RH) was infused into pentobarbitone-blocked pro-oestrous rats, it was concluded that (a) differences between the two types of preovulatory LH surges originate within the central nervous system, and (b) pro-oestrous LH-RH secretion may not be restricted to the period in which increased LH levels in blood were found, but may exceed this period for a considerable time.

Animals↗
PubMed1977

Sialic acid concentration in the reproductive organs, pituitary gland and urine of the Indian langur monkey (Presbytis entellus entellus).

The concentration of sialic acid was determined in the reproductive organs, pituitary gland and urine of male and female langur monkeys. In the male, sialic acid concentration was lowest in the testes and highest in the epididymides. Both caput and cauda epididymides contained almost the same concentration. Castration and subsequent androgen administration did not modify the concentration of sialic acid by comparison with intact animals, suggesting that sialic acid is not androgen-dependent in the langur. There was no significant difference in sialic acid concentration in the uterus during the proliferative and luteal phases of the menstrual cycle. After ovariectomy, there was a decrease in the concentration of sialic acid in the uterus, cervix and vagina while oestrogen or oestrogen plus progesterone administration increased its concentration significantly, but not to the control level. However, the difference in concentration between oestrogen-treated and oestrogen plus progesterone-treated monkeys was not significant. There was a peak of sialic acid excretion in the urine on day 8 of the normal 22-day menstrual cycle, i.e. a day before the probable day of ovulation. In pregnant monkeys siliac acid excretion increased significantly 2 days before delivery compared with the values recorded 7 to 3 days before parturition. The part played by sialic acid during the menstrual cycle and pregnancy in the female is not clear.

Animals↗
PubMed1984

Indices of ovulation: comparison of plasma and salivary levels of progesterone with urinary pregnanediol.

An increased daily excretion of pregnanediol, relative to that early in the menstrual cycle, is often taken to be evidence that a woman has ovulated. This paper assesses the value of alternative procedures for this purpose. Urine, plasma and saliva samples were collected during a 24-h period from 20 women during the follicular phase and from 20 women during the luteal phase. The 24-h excretion of pregnanediol was compared with (1) the concentration of progesterone in plasma, (2) the concentration of progesterone in saliva, (3) the concentration of pregnanediol in small urine samples, (4) the rate of excretion of pregnanediol and (5) the ratio of pregnanediol to creatinine in small urine samples. Each analyte increased substantially during the luteal phase. The median increases (ratio of luteal to follicular phase values) were 14.8, 3.2, 10.6, 11.9 and 11.1 respectively. By comparison, the median increase in 24-h pregnanediol output was 9.2. When the other analytes were used instead of the 24-h excretion of pregnanediol to assess the possibility of ovulation, the incidence of misclassifications (follicular samples classed as luteal and luteal samples classed as follicular) was 0, 12.8, 5.9, 2.0 and 1.0% respectively. It was concluded that the most satisfactory alternative to the measurement of 24-h pregnanediol output for the biochemical assessment of ovulation based on progesterone production was the measurement of the concentration of progesterone in plasma; the least satisfactory alternative was determination of the concentration of progesterone in saliva. If blood was not available, measurement of the ratio of pregnanediol to creatinine in a small urine sample was the preferred method.

Adult↗