Estradiol and progesterone concentrations resulting from the administration of an exogenous hormone regimen in diestrous embryo transfer recipients

Estradiol and progesterone concentrations were evaluated from diestrous embryo transfer recipient mares (5 to 14 days post-ovulation) which were treated with an exogenous hormone regimen. Upon detection of the donor mare's ovulation (0 hours), 10 mg PGF_2α was given to the recipient mare; at 12, 24 and 36 hours 20 mg estradiol cypionate; at 48 hours, 500 mg progesterone in oil and then 22 mg altrenogest at 60, 72 and 96 hours. Altrenogest (22 mg/day) was continued until end of the trial (detection of a fetal heart beat). Embryos were transferred non-surgically 6 or 7 days after the start of treatment.Plasma samples were evaluated over three periods; period 1-between recipient mare ovulation and prior to PGF_2α period 2-between PGF and embryo transfer and period 3-post-transfer. During periods 2 and 3, estradiol was higher (P<.05) for mares which were 10 to 14 days post-ovulation (late diestrous) as compared to mares which were 5 to 9 days post ovulation (mid-diestrous) when treatment began. Progesterone concentrations were higher (P<.05) for the mid-diestrous mares in the same periods. The pregnancy rate was higher for the late diestrous mares than the mid-diestrous mares (58% (7/12) vs 10% (1/10)). However, no difference (P>.05) was detected in estradiol or progesterone in the late diestrous mares which were pregnant or open. During period 2, estradiol was higher (P<.05) in the pregnant than open mares. Whereas, during period 3, progesterone was higher (P<.05) in the open mares.These data suggest that estradiol is important for the establishment of pregnancy in the mare. Furthermore, hormone treatment developed in this study appears to have some potential in synchronization of diestrus mares to be used as embryo recipients.     

Effect of progesterone supplementation on pregnancy and embryo survival in ewes

Two hundred eighteen ewes were used in experiments 1) to develop a progesterone supplementation regimen capable of sustaining serum concentrations of progesterone at about 2.0 ng/ml for a period of 50 d (Exp. 1) and 2) to determine the effects of progesterone supplementation (d 6 to 50 after mating) on pregnancy and embryo survival rates in mated ewes (Exp. 2). In ovariectomized ewes in Exp. 1, s.c. administration of four cylindrical (9.5 x 60 mm) silastic implants, containing 20% (1.1 g) progesterone by weight, sustained mean serum concentrations of progesterone of 1.9 +/- .07 ng/ml compared with 1.03 +/- .05 ng/ml in ewes bearing two implants. In Exp. 2 each ewe (n = 159) was mated to two fertile rams at a spontaneous estrus (d 0) during mid-breeding season. Mean ovulation rate, determined on a subgroup of 46 ewes, was 1.45 +/- .05. On d 6, ewes were assigned randomly to control (two implants containing no progesterone) or progesterone-treated (four implants similar to those used in Exp. 1) groups. From d 7 to 50 after mating, progesterone concentrations in serum were greater (P less than .001) in progesterone-treated (four implants similar to those used in Exp. 1) groups. From d 7 to 50 after mating, progesterone concentrations in serum were greater (P less than .001) in progesterone-treated (3.50 +/- .06) than in control (2.65 +/- .05) ewes. Pregnancy rates (86% and 83%) and calculated embryo survival rates (77% and 78%) were similar (P greater than .05) for the control and progesterone-treated groups, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pregnancy diagnosis in dairy goats and cows using progesterone assay kits

Early pregnancy diagnosis in dairy goats and cows was studied using enzyme immunoassays (EIA) to measure progesterone concentrations in whole milk samples collected approximately 3 weeks after mating. Two qualitative on-farm assay kits and 2 quantitative assay kits, all designed for use in the dairy cow, were tested for their accuracy with goats milk samples. Accuracy of diagnosis of goat pregnancy ranged from 83 to 88%, and of non-pregnancy from 80 to 100%. Pregnancy diagnosis with samples of cows milk using 2 quantitative kits gave accuracies of 66 to 68% for pregnancy, and 90 to 91% for non-pregnancy. Possible causes of error in the early diagnosis of pregnancy with milk samples are discussed.     

Use of synchronized heifer-recipients in view of the effectiveness of embryo transfer in cattle

A possibility was investigated of using heifers prepared for embryo reception by i. m. double application of Oestrophan (Spofa) luteolytic preparation. After a clinical examination of ovaries and corpus luteum, out of the total number of 2894 animals synchronized for nonsurgical ipsilateral transfer in the D7 stage only 2038 (70.4%) recipients were used. Almost 30% (856) heifers were discarded from the role of recipients, mostly due to the functional disorders of the cyclic activity of ovaries manifesting themselves as luteal cysts (22.5%), cysts of corpus luteum (16.5%), small or young corpora lutea (12.7%), postovulation states (4.5%), and also as a high frequency of the follicular activity without the presence of corpus luteum (37.9%). The occurrence of follicular cysts (1.4%) and nonfunctional ovaries (2.8%) was considerably lower. The high number of discarded recipients which results, in the complex of causes, from qualitative nutritional disorders, husbandry and zoo-hygienic shortcomings and stress-inducing factors in the course of recipient selection and preparation, diminishes the transfer efficiency and considerably increases the claims for the numbers of recipient prepared for ovulation. In the conditions of our workplace where 10.6 to 12.8 transferable embryos are recorded from one successful superovulation and their survival is 65 to 68.6%, it will be necessary, at the present level of recipient discarding, to prepare 13.8 to 16.6 heifers per donor to make full use of the superovulation effect.     

Calf removal improves conception rates to the Ovsynch and CO-Synch protocols

Beef cows (n = 473) from two locations were stratified by breed, postpartum interval, age, and AI sire and were randomly allotted to one of four treatments for synchronization of ovulation. Ovulation synchronization protocols included the Ovsynch protocol with (n = 114) or without (n = 123) 48-h calf removal from d 7 to 9 (d 0 = 1st GnRH injection) or the CO-Synch protocol with (n = 119) or without (n = 117) 48-h calf removal from d 7 to 9. The Ovsynch protocol included administration of GnRH (100 microg; i.m.) on d 0, PGF2alpha (25 mg; i.m.) on d 7, GnRH (100 microg; i.m.) on d 9, and timed insemination on d 10. The CO-Synch protocol included administration of GnRH (100 microg; i.m.) on d 0, PGF2alpha (25 mg; i.m.) on d 7, and GnRH (100 microg; i.m.) with timed insemination on d 9. Blood samples were collected from all cows on d -10 and d 0 for analysis of serum progesterone. Cows with at least one serum progesterone concentration greater than 1 ng/mL were considered to be cyclic at the time of treatment. Conception rates of cows that received the CO-Synch + calf removal, Ovsynch + calf removal, CO-Synch, or Ovsynch protocol (63, 61, 54, and 52%, respectively) were not different (P = 0.50). Conception rates were not different (P = 0.80) among CO-Synch- and Ovsynch-treated cows; however, both estrual status and 48-h calf removal affected conception rates. Conception rates of cyclic cows (66%) were greater (P = 0.01) than those of anestrous cows (53%), regardless of which synchronization protocol was used. When data were pooled across synchronization protocol, conception rates of cows with 48-h calf removal (62%) were greater (P = 0.09) than conception rates of cows without calf removal (53%). The CO-Synch + calf removal protocol induces a fertile ovulation in cyclic and anestrous cows, requires handling cattle just three times, results in high conception rates from timed insemination, and should be a useful program for synchronization of ovulation in beef cows.     

Effectiveness of intravaginal progesterone inserts and FSH for inducing synchronized estrus and increasing lambing rate in anestrous ewes

The objectives of this study were to determine whether a new progesterone (P4)-releasing intravaginal insert would induce fertile estrus and whether FSH combined with the insert would increase prolificacy in anestrous ewes introduced to rams. Ewes of mixed breeding on six farms were assigned to four randomized treatments: control (C), n = 73; 12 d P4 (polycapralactone [PCL] insert with 0.82 g P4), (P12), n = 73; 12 d P4 plus i.m. FSH (Folltropin, 55 mg NIH-FSH-P1 equivalent) in propylene glycol, 24 h before insert removal, (P12F), n = 71; and 5 d P4 plus FSH (P5F), n = 77. Growth and ovulation of follicles were observed ultrasonographically in 20 ewes at four farms (five/treatment) at insert removal and 36, 48, 72, and 96 h later. Intact rams (1:15 ewes in multiple-sire groups) were joined at insert removal, and raddle marks were observed every 12 h for 5 d. On d 26 to 30, rams were removed; ewes were examined for pregnancy then and 20 d later. Percentage of ewes marked by rams was greater in P4-treated (66 to 79%) than in C (12%; P < 0.01) ewes and in P5F (79%) than in P12F (66%; P < 0.05). Diameters of largest follicles at insert removal were greater (P < 0.05) in P4-treated (5.5 +/- 0.2) than in C ewes (4.8 +/- 0.2). Progesterone increased numbers of follicles > 3 mm (P < 0.01) or ovulated (P < 0.05; 2.6 +/- 0.6 vs 1.3 +/- 0.6 in C ewes) and FSH increased number of follicles > 3 mm (P < 0.05). In FSH-treated ewes, ovulation rate tended to be greater after treatment with P4 for 5 than for 12 d (P = 0.09, 3.3 +/- 0.6 and 2.2 +/- 0.4, respectively). More P4-treated than C ewes lambed (P < 0.01) to the first (38 to 45 vs 0%) or both (63 to 66 vs 41%) service periods. Prolificacy (first service) did not differ between FSH-treated ewes (P12F + P5F; 1.8 +/- 0.1) and ewes treated with P4 only (P12; 1.6 +/- 0.1). However, FSH increased prolificacy to first service (1.8 +/- 0.1) over prolificacy to second service (C ewes 1.5 +/- 0.1; P < 0.05, and all ewes 1.4 +/- 0.1; P < 0.01). Pregnancy retention did not differ among treatments but was greater (P < 0.01) in ewes that conceived at the first (90.9 +/- 3.7) than at the second (72.5 +/- 3.3) service period. In conclusion, a PCL insert in combination with ram introduction at insert removal was more effective than ram introduction alone to induce synchronized estrus and ovulation and to yield pregnancy after one or two service periods. Treatment with P4 for 5 d was as effective as for 12 d to induce fertile estrus in FSH-treated anestrous ewes.     

Blood types of twin cattle after embryo transfer to inseminated recipients

The blood types, together with erythrocyte and plasma protein types were determined in 10 pairs of twins born after embryo transfer to the contralateral uterine horns of previously inseminated recipients. These estimations were carried out at the age of nine weeks. According to the haemolytic test, most of these pairs of twins had identical blood types, ie, no erythrocyte mosaic could be demonstrated and the blood type corresponded to that of the recipient's calf. In the cases where mosaicism could be demonstrated, the recipient calf's blood type predominated. The reason for this bias remains unclear although the suggestion that the recipient's calf had developed further and was thus able to influence the haematopoietic tissues of its co-twin is worth considering.     

Day of embryo collection, quality and pregnancy rates in cattle

In 1300 donor cows, total embryos increased from 8.5 to 15.3, (P less than 0.001), the mean number of transferable embryos from 3.1 to 6.5 (P = 0.067) and pregnancies increased from 1.3 to 3.3 (P = 0.584) as the day of collection increased from six to 7.5. Most of the embryos recovered on day 6, 7.5 and 8 were morulae, early blastocysts and late blastocysts respectively. Morulae formed the majority of the embryos collected on days 6 and 6.5. Pregnancy rates in early and late blastocysts were highest on days 7 (54.4 and 60.2 per cent) and 7.5 (53.6 and 53.1 per cent, P = 0.009 and 0.004). There were significant differences in pregnancy rates between embryo stages on days 7 and 7.5 (P less than 0.001), embryo grades on days 7, 7.5 and 8 (P less than 0.001), and within embryo grades 1 (P = 0.015) and 3 and 4 (pooled, P = 0.009). On all days combined there were significant differences in pregnancy rates between both embryo stages (P = 0.007) and embryo grades (P less than 0.001). It appears that the concept of embryo fitness may be applied to blastocysts but not to morulae.     

Effect of oestrous synchrony between embryo donors and recipients,embryo quality and state on the pregnancy rate in beef cattle

This study aimed to evaluate the effect of oestrous synchrony between donors and recipients and the embryo quality on the pregnancy rate in beef cow recipients. The experiment was performed over two years at an embryo transfer (ET ) centre in Southern Brazil. Ninety Aberdeen Angus cows were subjected to superovulation (SOV ) protocols, resulting in the recovery of 1,048 transferable embryos. Eleven groups were formed with intervals of 6 hr, from ?30 to +30 hr, with respect to recipient versus donor oestrous detection. Evaluation of embryo quality was according to the IETS guidelines. The overall pregnancy rate was 52%. Effects related to donor and recipient oestrous synchronization on pregnancy rate were observed (p  =  .01), ranging from 36% to 50%. The embryo quality rate affected the pregnancy rate, where Grade I resulted in 57% and Grade III in 43% of pregnancy (p  <  .001). The embryonic state (frozen or fresh) showed no (p  >  .05) effect on pregnancy rate: 53% for fresh embryos and 44% for frozen embryos. The odds ratio for explanatory variables causing pregnancy indicated that Grade III embryos had 31% less chance of conception compared to Grade I. Thus, oestrous synchrony between donor and recipient, considering ±30 hr apart, can affect the pregnancy rate along with embryo quality.     

Effect of estrus synchronization protocols on plasma progesterone profile and fertility in postpartum anestrous Kankrej cows

The study was aimed at induction/synchronization of estrus in postpartum anestrous Kankrej cows of zebu cattle maintained at an organized farm. The study included use of different hormone protocols, viz., Ovsynch, CIDR (controlled internal drug release), Ovsynch plus CIDR, and Heatsynch with estimation of plasma progesterone on days 0, 7, 9/11 (artificial insemination--AI) and on day 20 post-AI following fixed time insemination. Thirty selected anestrous animals were divided into five equal groups (four treatment and one control), and the findings were compared with the normal cyclic control group of six cows. All the protocols were initiated in cows with postpartum anestrous period of more than 4 months, considering the day of first GnRH injection or CIDR insertion as day 0. The animals were bred by fixed time artificial insemination. Pregnancy was confirmed per rectum on day 60 post-AI in non-return cases. The conception rates at induced/first heat in Ovsynch, CIDR, Ovsynch + CIDR, and Heatsynch protocols were 33.33, 66.66, 50.00 and 16.67%, respectively. The corresponding overall conception rates of three cycles post-treatment were 50.00% (3/6), 100.00% (6/6), 66.66% (4/6), and 50.00% (3/6). In normal cyclic and anestrous control groups, the pooled pregnancy rates were 83.33% (5/6) and 16.67% (1/6), respectively. The pooled mean plasma progesterone (nanograms per milliliter) concentrations were significantly (P < 0.05) higher on day 7 in Ovsynch (5.727 ± 1.26), CIDR (4.37 ± 0.66), Ovsynch plus CIDR (3.55 ± 0.34), and Heatsynch (5.92 ± 1.11) protocols as compared with their corresponding values obtained on days 0, 9/11 (AI), and on day 20 post-AI. In anestrous control group, the mean progesterone concentration at the beginning of experiment was 0.67 ± 0.33 ng/ml, which was at par with values of all other groups. The overall plasma progesterone levels on the day of initiating treatment were low in all groups, with smooth small inactive ovaries palpated per rectum twice at 10 days interval, suggesting that most of the animals used in the study were in anestrous phase. Mean (± SE) values of plasma progesterone (nanograms per milliliter) on day 20 post-AI were higher in conceived cows than the non-conceived cows of all the groups, but differed significantly (P < 0.05) only in normal cyclic group. These results suggest that use of different hormone protocols particularly Ovsynch, CIDR, and Ovsynch + CIDR may serve as an excellent tool for induction and synchronization of estrus and improvement of conception rate in postpartum anestrous Kankrej cows.     

Relationship between pregnancy rate and serum progesterone concentration in cases of porcine embryo transfer

The level of P4 at the time of embryo transfer (ET) is important. P4 concentrations and numbers of corpora lutea for 126 recipients were evaluated. Nuclear transfer embryos were transferred into 126 surrogates. 11 maintained their pregnancy until full-term delivery, 17 miscarried, and implantation failed in 98 animals. P4 levels in the full-term group were significantly different from those of the pigs that aborted or in which implantation failed (p < 0.05). However, the numbers of corpora lutea were not significantly different. These findings indicate that the concentration of progesterone can be an important factor for successful ET in pigs.     

家兔超数排卵和胚胎移植妊娠率的影响因素分析

应用FSH和HCG相结合的不同方法 (递减量与恒量法),对不同兔龄和是否经产的中国家兔进行了超数排卵处理。其中,递减量法超排处理:每间隔12 h依次肌肉注射15、15、10、10、5及5 U/只共60 U的FSH,6次之后间隔12 h,经耳缘静脉注射HCG 100 U/只,合笼;恒量法超排处理:每间隔12 h肌肉注射10 U/只共60 U的FSH,6次之后间隔12 h,经耳缘静脉注射HCG 100 U/只,合笼。17 h后,取卵经显微注射后移植到排卵点分布不同的家兔输卵管中,比较分析了超数排卵及胚胎移植妊娠率的影响因素,旨在优化兔超数排卵和胚胎移植方法。结果显示,①总量为60 U的FSH递减量肌肉注射法超排效果略优于恒量法(688枚、671枚),但差异不显著(P>0.05);②9～11月龄的母兔超排效果明显优于6～8月龄的母兔(794枚、565枚),差异显著(P<0.05);③经产母兔的超排效果明显优于未经产的母兔(887枚、472枚),差异显著(P<0.05);④经显微注射重组DNA的胚胎移植受体时,两侧卵巢均有排卵点的胚胎移植受体妊娠率明显高于一侧卵巢有排卵点和两侧卵巢均无排卵点的受体(81.25%、38.46%、37.50%),差异显著(P<0.05)。试验证明,通过肌肉注射递减总量为60 U的FSH对9～11月龄经产母兔的超排效果最佳,且两侧卵巢均有排卵点的受体胚胎移植妊娠率最高,为胚胎研究工作者提供了一定的参考依据。     

The comparison of treating Holstein dairy cows with progesterone, CIDR and GnRH after insemination on serum progesterone and pregnancy rates

This experiment was conducted to investigate the effects of different exogenous progesterone sources administrated after artificial insemination (AI) on serum progesterone (P(4) ) concentration and pregnancy rates in Holstein lactating cows. Sixty-four lactating Holstein dairy cows were allocated to four different treatments (n = 16 per treatment): the cows 1) were injected with physiological saline on days 5 and 13 after AI (control group); 2) were injected with progesterone on days 5 and 13 after AI (P group); 3) received controlled internal drug releasing device (CIDR) for a period from day 5 to 19 after AI (CIDR group); and 4) were injected with gonadotropin-releasing hormone (GnRH) agonist on days 5 and 13 after AI (GnRH group). Blood samples were collected on days 0 (AI day), 5, 13, 16 and 19 after AI to determine serum P(4) concentration. The results revealed a significant difference among treatment groups for serum P(4) concentration on days 13, 16 and 19 with the lowest concentration of serum P(4) for the control group. The pregnancy rate was also positively affected by all the treatments with CIDR having the greatest effect on pregnancy rate. Overall, the results indicated that CIDR has the greatest effect on serum P(4) concentration and pregnancy rate, although the administration of P and GnRH during days after AI increased serum P(4) concentration in lactating dairy cows as well.