Folkmar Elsaesser

Pregnancy rates relative to recipient plasma progesterone levels on the day of nonsurgical transfer of frozen/thawed bovine embryos

A total of 71 synchronized dairy heifers (Holstein Friesian x German Black Pied) were used as recipients of seven-day old frozen/thawed bovine embryos. Plasma progesterone concentrations and corpus luteum quality on the day of nonsurgical transfer (= day 7) were determined and related to pregnancy rates or estrus intervals in nonpregnant recipients. A total of 32 recipients (45.1 %) maintained pregnancy; 39 recipients (54.9 %) did not. No significant differences could be detected between progesterone levels in recipients that remained pregnant (3.14 +/- 0.24 ng/ml; x +/- SEM ) and those that did not maintain pregnancy (3.23 +/- 0.28 ng/ml). Optimal progesterone levels were between 2 and 5 ng/ml coinciding with a pregnancy rate of 51.1 % (24 47 ). Pregnancy rates apparently were decreased when progesterone levels were below 2 ng/ml (35.3 %; 6 17 ) or above 5 ng/ml (28.6 %; 2 7 ). Hence, optimal progesterone levels were identical to those for freshly collected embryos reported previously by Remsen et al. (1). Bovine corpus luteum quality graded by rectal palpation was related to some extent to progesterone levels but not to pregnancy rates. Out of 39 nonpregnant recipients seven animals (17.9 %) with a mean plasma progesterone level of 3.76 +/- 0.72 ng/ml showed an extended estrus interval of more than 55 days, probably indicating early embryonic mortality. Progesterone levels did not significantly differ between nonpregnant recipients with estrus intervals of various length. Plasma progesterone levels at the time of transfer are of limited diagnostic value for screening recipients prior to transfer of frozen/thawed embryos.

Superovulation of dairy cows with purified FSH supplemented with defined amounts of LH.

This study investigated the effects of a purified follicle stimulating hormone (FSH) preparation supplemented with three different amounts of bovine luteinizing hormone (bLH) and a commercially available FSH with a high LH contamination on superovulatory response, plasma LH and milk progesterone levels in dairy cows. A total of 112 lactating Holstein-Friesian crossbred dairy cows were used for these experiments; the cows were randomly assigned to treatment groups consisting of purified porcine FSH (pFSH) supplemented with bLH. Group 1 was given 0.052 IU LH/40 mg armour units (AU) FSH (n = 6); Group 2 was given 0.069 IU LH (n = 32); Group 3 received 0.423 IU LH (n = 34); while Group 4 cows (n = 36) were superovulated with a commercially available FSH-P. This compound appeared to contain 8.5 IU LH/40 mg AU FSH according to bioassay measurement. All animals received a total of 40 mg AU FSH at a constant dose twice daily over a 4-d period. Levels of milk progesterone and plasma LH were determined during the course of superovulatory treatment. The Group 1 treatment did not reveal multiple follicular growth, and no embryos were obtained. Superovulation of Group 3 cows resulted in significantly (P<0.05) more corpora lutea (CL; 12.6+/-1.1) and fertilized ova (5.1+/-1.3) compared with Groups 2 and 4 (10.1+/-0.9 and 2.6+/-0.6, 9.0+/-0.9 and 2.7+/-0.5, respectively). Due to a high percentage of degenerated embryos (33%) Group 3 yielded only one more transferable embryo than Groups 2 and 4. Among groups, LH levels differed in the period prior to induction of luteolysis and were similar thereafter. The progesterone pattern following FSH/LH administration reflected the amount of LH supplementation. Milk progesterone levels on the day prior to embryo collection were correlated to the number of CLs and recovered embryos. It is concluded that under the conditions of our experiment superovulation with 0.423 IU LH/40 mg AU FSH may yield a significantly improved superovulatory response in dairy cows. It is further suggested that LH supplementation exerts its effects mainly on follicular and oocyte maturation during the period prior to luteolysis.

Prospects and problems of pregnancy and fertility diagnosis in the pig by aid of progesterone determination.

Plasma progesterone levels were measured in 536 sows of different breeds from 6 different farms to assess this test as a method of determining early pregnancy. It was found that progesterone curves of pregnant and nonpregnant animals began to diverge on Day 16 after mating; nonpregnant animals who turned out to have a prolonged cycle could be eliminated by testing on Days 17-24. From Day 17 on only 1 pregnant animal showed values lower than 9 ng progesterone/ml plasma while 86% of all nonpregnant animals had values below 5 ng. 3.6% of animals were incorrectly diagnosed on the basis of plasma progesterone .4% of pregnant animals considered nonpregnant and the rest nonpregnant animals diagnosed as pregnant mainly due to a delayed return of estrus. A 2nd study found that sows with values below 9 ng on Days 10-15 had a history of infertility. Very high progesterone levels (110-144 ng/ml far above those normally observed in pregnant animals) were not associated with any functional disturbances. The method shows promise as a means of fertility diagnosis and can also be used to detect animals with weak or no behavioral estrus.

Rapid milk progesterone assay as a tool for the selection of potential donor cows prior to superovulation

This study investigated the accuracy of a commercially available rapid milk progesterone (P(4)) assay (RMPA) and its usefulness for the screening of potential donor cows prior to superovulatory treatment. Superovulation was induced in 90 lactating Holstein-Friesian crossbred dairy cows with twice daily injections over a 4-day period for a total of 40 mg follicle stimulating hormone (FSH-P), starting 9 to 13 d post estrus. Prior to induction of superovulation, a milk sample was collected and assayed for a P(4) level using the RMPA. The test determines P(4) by a simple visual color inspection of the respective sample, which is compared to a standard containing 10.5 ng/ml of P(4). All animals were divided into six groups according to the color intensity of their sample; three groups had a lower level, one group had an equal level and two groups had a higher P(4) level than the standard. Results of the semiquantitative RMPA were verified by a quantitative enzymeimmunoassay (EIA). Samples evaluated as equivalent to the standard had a mean P(4) level of 10.7 +/- 1.3 ng/ml (x +/- SEM). In total, P(4) levels differed (P<0.05) among groups, except in those with lower P(4) concentrations (1.1 +/- 0.0; 1.0 +/- 0.0; 3.7 +/- 1.5; 10.7 +/- 1.3; 13.8 +/- 1.3; 19.0 +/- 1.5 ng/ml, respectively). The correlation between RMPA-groups and EIA P(4) levels was 0.69 (P<0.001). Donors classified as having less P(4) than the standard yielded fewer corpora lutea (CL) (P<0.005), ova and embryos (P<0.05), and transferable embryos (P<0.05) compared with donors having similar or higher P(4) levels (3.4 +/- 1.0 vs 10.8 +/- 0.7 CL; 1.7 +/- 0.8 vs 6.2 +/- 0.9 ova and embryos; 1.2 +/- 0.7 vs 2.8 +/- 0.4 transferable embryos). Our results indicate that RMPA determines milk P(4) levels with sufficient accuracy and is a simple and useful tool for the screening of potential donor cows.

In vitro characterization of secretion rates from silastic micropellets containing estradiol

For local, controlled steroid hormone administration into tissues, such as the brain, we have prepared cylindric micropellets of 1 mm in length and 1 mm diameter. The micropellets are a mixture of silicone glue (silastic) and 0.1%, 1.0%, or 10% estradiol (E2). To evaluate in vivo E2 secretion rates, micropellets were implanted into the brains of 40 rats for either 1, 4, 8, or 12 weeks. In vitro 24 h E2 secretion rates of these implants were compared-after removal from the rat brain-with 24 h secretion rates of micropellets that had been incubated for the same periods of time in vitro only. In vitro release of E2 decreased steadily but asymptotically from the first day of incubation to the 3rd or 4th week, when an apparent steady state is achieved. With any E2 concentration the coefficient of variation for 24 h release rates rarely exceeded 15% within a group. The release rates increased nonlinearly with the concentration of E2 in the pellet. Subsequent to in vivo implantation the in vitro secretion of E2 was slightly higher than the in vitro secretion of micropellets incubated for the same period of time in vitro. Thus (1) the secretion rate from a pellet can be predicted rather exactly by the mixing ratio of silastic and E2 and (2) the secretion rate from the micropellet in vitro and in vivo appears to be rather similar. It is concluded that the method described is very useful for short-term (days) or long-term (weeks, albeit not constant) local exposure of defined tissues to steroid hormones.