scholarly journals Influence of Injectable Progesterone on the Pregnancy Rate of Heifers Receiving Bovine Embryos

2020 ◽  
Vol 48 ◽  
Author(s):  
Caio Cezar Da Silva ◽  
Hélton Aparecido Garcia Gregianini ◽  
Jennifer Teodoro Ferreira Gregianini ◽  
José Antônio Dell’Aqua Junior ◽  
Jefferson Viana Alves Diniz ◽  
...  

Background: In vitro embryo production (IVEP) allows the spread of superior animal genetics, but pregnancy rates show a high variability with this biotechnique. In the initial stage of pregnancy, progesterone plays a fundamental role in uterine preparation, acting on embryonic growth, implantation, and development. However, on the day of the IVEP transfer to the recipients, progesterone levels may be lower than that expected, influencing the uterine environment and, consequently, the pregnancy rate. Therefore, the objective of this study was to evaluate the pregnancy rate in heifers after the administration of injectable progesterone (P4) in the fixed-time embryo transfer (FTET) protocol.Materials, Methods & Results: The experiment was conducted inside a rural property near the city of Rio Branco, Acre, Brazil. The experimental group consisted of 232 animals, including 78 zebuine (Bos indicus) and 154 mixed (½ blood B. indicus and ½ blood B. taurus) animals, aged between 16 and 24 months, with a mean weight of 300 and 330 kg for zebuine and mixed animals, respectively. The selected animals were previously synchronized using the progesterone-estrogen-prostaglandin-estrogen protocol. Embryo transfer was performed on day 18 of the protocol, which was 9 days after the removal of intravaginal progesterone implant. On day 15 of the protocol, that is, 144 h (6 days) after the device removal, the animals were randomly distributed into two experimental groups: Control Group (CG; 0.5 mL of 0.9% saline solution, intramuscular) and Treated Group (P4G; 0.5 mL of injectable P4, 150 mg, intramuscular). Chi-square test was used for the statistical analysis of the pregnancy rate at a 5% probability. After 23 days of embryo transfer, pregnancy was diagnosed by ultrasonography. The general pregnancy rate, considering all groups (CG and P4G) and breeds included, was 55.17% (128/232). The pregnancy rates of the P4G and CG groups, regardless of breeds, were 55.08% (65/118) and 55.26% (63/114), respectively, with no statistical difference (P = 0.8344). Angus animals presented a similar pregnancy rate in the P4G and CG groups of 54.93% (39/71) and 57.83% (48/83), respectively. Furthermore, similar results were found for the Nellore breed, with no difference in pregnancy rate between the CG (55.81%, 24/43) and P4G (48.57%, 17/35) groups.Discussion: The strategy of using P4 to increase the pregnancy rate is very widespread in fixed-time artificial insemination (FTAI) protocols, although differences still exist. The literature presents zero or negative effects, as in this study, when exogenous P4 was used to increase endogenous P4 concentrations and, consequently, the pregnancy rate. However, some studies demonstrated the beneficial effects of increasing blood P4 concentrations, and that the increased fertility depends on the method and time of P4 supplementation and the animal’s physiological state. In this context, the fact that the injectable progesterone supplementation did not increase the pregnancy rate in this study is justified mainly by factors such as the animal category used (heifers), time of P4 supplementation (day 4 after ovulation), biotechnology used (FTET), and the heterogeneous characteristics related to phases of the estrous cycle of heifers. Supplementation with 150 mg of injectable long-acting progesterone intramuscularly did not interfere in the pregnancy rate of Angus and Nellore heifers receiving bovine embryos.

2004 ◽  
Vol 16 (2) ◽  
pp. 212 ◽  
Author(s):  
T. Nishisouzu ◽  
M. Sugawara ◽  
S. Aoki ◽  
K. Kishida ◽  
M. Moriyoshi ◽  
...  

Treatments with GnRH and PGF2α for synchronization of ovulation has resulted in acceptable pregnancy rates after fixed-time artificial insemination in dairy cows without estrus detection. The objective of the present study was to evaluate the practicability of ovulation synchronization (Ovsynch, Pursley JR et al. 1995 Theriogenology 44, 915–923) in dairy cattle using GnRH and PGF2α for the embryo transfer recipients. Dairy cattle (cows; n=100, heifers; n=88) were randomly allocated to one of two groups. The control group (cows; n=45, heifers; n=37) was composed of cows in natural estrus. The ovulation synchronization group (cows; n=55, heifers; n=51) was treated with an intramuscular injection of 100μg of GnRH at a random stage of the estrous cycle. Seven days later, the cattle received PGF2α (Cows; 25–30mg) or PGF2α analog (Heifers; 0.5mg) in order to regress the corpora lutea (CL). Forty-eight hours later, cows and heifers received a second injection of 100μg GnRH. Embryo transfer was carried out 7 days after the second injection of GnRH in the ovsynch group and 7 days after estrus in the control group. The cattle judged to have CL 17mm were classified as acceptable recipients. The size of the follicles and the CL were determined to be of estrus stage and embryo transfer by means of ultrasonography. The mean numbers of follicles and CL were analyzed by ANOVA, while pregnancy rates were analyzed by chi-square test. The results are presented in the Table. The proportion of cows and heifers determined to be acceptable embryo transfers was not different between the control group and the ovsynch group. There were no differences in the proportion of acceptable embryo transfers between the control group and the ovsynch group. Follicle diameter at the time of estrus in the control group (cows; 20.7±0.7mm, heifers; 16.8±0.5mm) were significantly larger than that of the ovsynch group (cows; 18.0±1.0mm, heifers; 14.7±0.2mm) (P<0.05). Although CL diameter at the time of embryo transfer in heifers showed no differences between the control group and the ovsynch group (25.0±1.0mm v. 22.8±1.5mm), The CL diameter of the control cow group was larger than that of the ovsynch group (29.8±0.7mm v. 26.1±1.0mm, P<0.05). However, no differences in pregnancy rate were seen between the control group and the ovsynch group. These results suggest that ovsynch can be effectively applied in an embryo transfer program for cattle. Table 1 Proportion of acceptable embryo transfer recipients and pregnancy rate in dairy cattle in the control ovsynch groups


2014 ◽  
Vol 26 (1) ◽  
pp. 163
Author(s):  
C. A. C. Fernandes ◽  
J. R. Ribeiro ◽  
M. P. Palhao ◽  
A. C. S. Figueiredo ◽  
J. H. M. Viana ◽  
...  

In most Brazilian farms, the management of embryo recipients is done exclusively under grazing systems. Thus, the seasonal variation of grass availability can affect the energy balance and reduce weight gain, and consequently affects the rate of pregnancy. The aim of the present study was to evaluate the effect of the seasonal variation of daily weight gain (DWG) on the pregnancy rate of crossbred heifers (Bos taurus × Bos indicus). The study was performed in the southwest region of Minas Gerais State, Brazil, during one year. Two periods were evaluated: dry (April to September, average precipitation of 470 mm and low grass availability) and rainy (October to March, average precipitation of 1930 mm and high grass availability) seasons. The recipients were kept in a grazing system (Brachiaria brizantha) with water and minerals available ad libitum. Embryos were produced in vitro by the same laboratory, using oocytes and sex-sorted semen from the Gir breed (Bos indicus). A total of 404 fresh embryos (N = 484, 45.4% during dry and 54.5% during rainy season) were transferred by the same technician on Day 7 days of the oestrous cycle – synchronized with prostaglandin – of the recipients, which were weighted at the day of embryo transfer and 23 to 25 days later, at the diagnostic of pregnancy. Animals were retrospectively allocated into 4 groups according to the DWG observed between embryo transfer and pregnancy diagnosis: G1 (N = 132) up to 150 g; G2 (N = 132) between 151 and 250 g; G3 (N = 119) between 251 and 350 g; and G4 (N = 99) greater than 350 g of DWG. The pregnancy rates were compared among DWG groups and between seasons by chi-squared. The logistic regression model was performed using the PROC GENMOD to test the model, including the effects of DWG, season, and the interaction (DWG × season). During the dry season, there was change in frequency distribution of the heifers among DWG groups, with lower frequency (P < 0.05) of heifers in high (G3 and G4) when compared with low (G1 and G2) DWG groups (33.2, 31.4, 21.8, and 13.6% for G1, G2, G3, and G4, respectively). Altogether, pregnancy rate was not different between dry and rainy seasons (42.3 v. 45.8%, respectively; P > 0.05). When compared to G1, pregnancy rate was higher (P < 0.05) for recipients with daily weight gain greater than 250 g day–1 (G3 and G4) (35.6, 40.6, 52.1, and 51.0% for G1, G2, G3, and G4, respectively). Only the effect of the DWG was significant in logistic regression, and the odds rate for each gram of DWG was 0.0039, which means that pregnancy rate improved 1.17 times for recipients with 300 g of DWG. In conclusion, the DWG after embryo transfer, but not the season itself, affect pregnancy rate of in vitro-produced embryo recipients. Biotran, FAPEMIG, and CnPQ are acknowledged.


2010 ◽  
Vol 22 (1) ◽  
pp. 170 ◽  
Author(s):  
L. A. Lima ◽  
V. G. Pinheiro ◽  
J. R. Cury ◽  
C. M. Barros

There are reports in the literature indicating that use of eCG improves pregnancy rates in Bos indicus anestrous cows treated with a progesterone-based fixed-time AI (FTAI) protocol. More recently, replacement of eCG by FSH in FTAI protocols was reported to have beneficial effects (Bos indicus) or no effect (Bos taurus) on pregnancy rates. In the present experiment the effects of eCG and FSH on pregnancy rates were compared in an FTAI protocol. Primiparous lactating Nellore cows (40 to 80 days postpartum, n = 421) with a body condition score of 2.5 to 3.0 (on a 1 to 5 scale) were randomly allocated in 3 groups: Control (CTR), eCG, and FSH. In the control group, all animals received a progesterone (P4)-releasing intravaginal device (1.55 g, PRID®, Ceva Sante Animale S.A., Libourne, France) and 2.5 mg of estradiol benzoate (EB, i.m. Estrogin®, Farmavet, São Paulo, Brazil), on Day 0 (D0). Eight days later (D8), at the time of the intravaginal device withdrawal, PGF2α (150 μg, D-cloprostenol, i.m. Prolise®, ARSA S.L.R., Buenos Aires, Argentina) was administered. Twenty-four hours after PRID removal, cows were treated with EB (1.0 mg, i.m.), and FTAI was done 30 to 36 h later. In the eCG and FSH groups, the cows were treated with 20 mg of FSH (Folltropin-V®, i.m. Bioniche, Belleville, Canada) or400IU of eCG (Novormon®, i.m. Syntex, Buenos Aires, Argentina), respectively, at the time of PGF2α administration. Ovarian ultrasonography (Aloka SSD 500, 7.5-MHz probe, Aloka, Tokyo, Japan) was performed 10 days prior to and at the beginning of FTAI protocol to select the animals in postpartum anestrous (absence of CL in both examinations). The pregnancy diagnosis was performed by ultrasonography 30 days after FTAI. Data were analyzed by logistic regression (SAS Institute, Cary, NC, USA). Cows treated with eCG had a higher pregnancy rate (41.5%, 95/229; P < 0.05) than those treated with FSH (22.7%, 22/97) or in the control group (26.3%, 25/95). Pregnancy rates of animals treated with FSH did not differ (P > 0.05) from those in the control group. These results indicated that, in lactating primiparous anestrous Nellore cows, the use of eCG in a FTAI protocol improves the pregnancy rate, whereas FSH has no beneficial effect. V. G. Pinheiro received a fellowship from FAPESP (São Paulo, Brazil). The authors are grateful to Ceva Animal Health for providing the intravaginal devices (PRID®) used in this experiment.


2016 ◽  
Vol 28 (2) ◽  
pp. 184
Author(s):  
M. Pelizzari ◽  
A. Tribulo ◽  
J. Garzon ◽  
B. Bernal ◽  
R. Tribulo ◽  
...  

A retrospective analysis of factors that affect pregnancy rates from 4214 fresh in vitro-produced (IVP) embryos that were transferred at a fixed-time (FTET) in 20 different farms. Recipients were all cycling cows or heifers that were synchronized with 1 of 3 treatments: 1) treatments with progesterone (P4) devices and 2 mg of oestradiol benzoate (EB) on Day 0 (day of insertion) and 24 h after device removal (Day 8); 2) treatments with P4 devices and EB on Day 0, but with 0.5 mg of oestradiol cypionate (ECP) at device removal (Day 8); or 3) treatments with P4 devices and GnRH on Day 0 and a second GnRH 60 h after device removal (Day 5). Cows in all treatment groups also received 500 µg of cloprostenol (prostaglandin F2α) at the time of P4 device removal and 400 IU of eCG either at device removal or 3 days before device removal. All embryos were transferred 7 or 8 days after the expected time of oestrus (24 h after EB, 48 h after ECP or at the time of the second GNRH for each synchronization treatment, respectively). On the day of embryo transfer, recipients were examined by ultrasonography and those with corpus luteum >14 mm in diameter received a fresh, IVP embryo in the uterine horn ipsilateral to the corpus luteum. Pregnancy rates were determined by ultrasonography 35 days after FTET. Data were analysed by logistic regression. Independent variables were classified into the following three categories. 1) Factors related to the recipient and the environment; there were no significant differences in pregnancy rates for corpus luteum diameter (≥14 and <16 mm, ≥16 and <18 mm, or ≥18 mm; P = 0.46), number of corpus luteum (1 or ≥2; P = 0.26), and category of recipient (cow or heifer; P = 0.21). However, there were significant effects of farm (P = 0.01) and body condition score (BCS; P = 0.01). Cows with BCS ≥4.5 (1 to 5 scale) resulted in lower pregnancy rates (4/20, 20.0%) than those with BCS 2 (74/225, 32.9%), 2.5 (502/1434, 35.0%), 3 (570/1467, 38.9%), 3.5 (193/532, 36.3%), and 4 (44/118, 37.3%). 2) Factors related to the synchronization treatment; there were no significant differences between recipients receiving eCG at device removal (84/209, 40.2%) or 3 days before device removal (874/2291, 38.1%; P = 0.35). However, recipients synchronized with P4 devices and ECP had higher (P = 0.01) pregnancy rates (232/483, 48.0%) than those treated with EB (679/1888, 36.0%) or gonadotropin-releasing hormone (47/129, 36.4%). 3) Factors related to the embryo transfer technique; day of the recipient’s oestrous cycle (P = 0.36), stage of embryo transferred (IETS stages 6 or 7; P = 0.62), and operator (P = 0.57) did not affect pregnancy rates. However transfers made in the anterior third of the uterine horn resulted in higher (649/1545, 42.0%) pregnancy rates than those in the mid-third (845/2511, 33.6%) or in the distal third (6/35, 17.1%; P = 0.01). It was concluded that factors related to the recipient and the environment (farm and BCS), the synchronization treatment (ECP), and the embryo transfer technique (site of deposition) affect pregnancy rates in recipients of embryos produced in vitro and transferred at a fixed time.


2014 ◽  
Vol 26 (1) ◽  
pp. 119
Author(s):  
J. B. S. Borges ◽  
D. X. Thedy

Successful establishment of pregnancy in the cow depends on early rise of progesterone (P4) to improve embryo development. The present study was carried out to determine the corpus luteum (CL) function and pregnancy rate following the administration of 1.500 IU of hCG in suckled beef cows on Day 4 after fixed-time AI (FTAI). Crossbred Angus cows (n = 429), 40 to 70 days postpartum, were synchronized using 2 mg of oestradiol benzoate (EB) i.m. and an intravaginal progesterone-releasing device containing 1 g of P4 for 7 days. At device removal, prostaglandin F2a (150 mg of D-cloprostenol) was injected i.m.; 24 h later, 1 mg of EB was given and the FTAI were done 30 to 36 h after EB using semen of just 1 bull. Cows were assigned alternatively to receive 1.500 IU of hCG i.m. (n = 200) or 1 mL of saline i.m. (n = 229) on Day 4 after FTAI. Ovaries were examined by transrectal ultrasonography (n = 71) on Days 4 and 7 to detect the presence and to measure the area of CL. Blood samples were collected on Days 4, 7, 10, 14, and 30 post-oestrus to determine P4 concentration by RIA. Pregnancy diagnoses were done 30 days after FTAI. The effects of treatments on CL area and serum P4 levels were analysed by ANOVA (Proc GLM, SAS Institute Inc., Cary, NC, USA) and pregnancy rates compared by chi-square (Proc CATMOT, SAS). The induction of accessory CL determined on Day 7 was 44% in cows treated with hCG and not observed in the control group. The area of the CL on Day 7 was significantly higher (P < 0.01; 3.37 ± 2.56 v. 2.56 ± 0.22 cm2) in the hCG group. Despite the increase in CL size, the mean levels of P4 on Days 4, 7, 10, 14, and 30 post-AI indicated no difference (P > 0.05) in CL function between control and hCG-treated cows. On Day 30, pregnancy rates were not different (P = 0.08), at 46.7 and 55% for control and hCG-treated cows, respectively. In conclusion, administration of 1.500 IU of hCG on Day 4 post-FTAI did not improve pregnancy rate in suckled beef cows.


2011 ◽  
Vol 23 (1) ◽  
pp. 163 ◽  
Author(s):  
R. Núñez ◽  
T. de Castro ◽  
L. Cutaia ◽  
G. Bó ◽  
A. Menchaca

An important proportion of embryonic deaths in cattle occur during the critical period of embryo development and maternal recognition of pregnancy. High serum progesterone concentrations during this period are associated to higher embryo survival, and the use of eCG could enhance luteal activity. Two experiments were carried out to evaluate the effect of administering eCG at the time of removal of a progesterone-releasing device and 14 days after fixed-time artificial insemination (FTAI) in beef cattle. Multiparous suckled cows between 60 and 90 days postpartum (n = 297; Exp. 1), and two-year old prepuberal heifers (n = 689; Exp. 2) with a body condition score (BCS) between 3.5 and 5 (1 to 8 scale) were used. At the beginning of the experiments, none of females used had a corpus luteum detected by ultrasonography. Females received an intravaginal device containing 0.5 g progesterone (DIB 0.5, Syntex, Buenos Aires, Argentina) and 2 mg oestradiol benzoate i.m. (Benzoato de Oestradiol, Syntex). Eight days later, 500 μg of cloprostenol (Ciclase DL, Syntex) and 0.5 mg oestradiol cypionate (Cipiosyn, Syntex) were given i.m. and devices were removed; FTAI was performed 52 to 56 h after DIB removal. At the time of device removal, the animals were randomly allocated to 4 groups, to receive 400 IU of eCG i.m. (Novormon, Syntex) or placebo at device removal and/or 14 days after FTAI, in a 2 × 2 factorial design. Control group did not receive eCG. In Exp. 1, nose plates were placed on calves on the day of DIB insertion and left for 10 days in order to prevent suckling. Pregnancy diagnosis was performed by ultrasonography at 30 and 60 days after FTAI. Data were analysed by logistic regression. Pregnancy rate at 30 days after FTAI was affected by treatment (P < 0.05; Table 1). Late embryonic/fetal loss rates (non-pregnant females at 60 days after FTAI over those pregnant at 30 days) were 1.6% (3/182) for cows in Exp. 1 and 4.8% (14/292) for heifers in Exp. 2; no differences were detected among groups. In conclusion, administration of 400 IU of eCG at device removal increases pregnancy rate, both in anestrus cows and heifers. However, the administration of a second dose of eCG 14 days after FTAI suggests a possible effect on embryo survival within the first 30 days of pregnancy in cows but not in heifers. Further studies are required to confirm these results. Table 1.Pregnancy rates after administration of eCG at time of progesterone device removal and/or 14 days after insemination in beef cattle Supported by Syntex.


2004 ◽  
Vol 16 (2) ◽  
pp. 212
Author(s):  
L.F. Nasser ◽  
E.L. Reis ◽  
A.M. Oliveira ◽  
G.A. Bo ◽  
P.S. Baruselli

It has been shown recently that treatments with progesterone (P4)-releasing devices combined with estradiol benzoate (EB) plus P4 on Day 0, eCG and PGF on Day 5 and a second application of EB one day after device removal (Day 9) can be used successfully to transfer bovine embryos at a self-appointed time, without the necessity of estrus detection. Although the treatment solved one of the major problems in recipient management, estrus detection, it requires handling the recipients at least five times for treatments and embryo transfer. An experiment was designed to evaluate whether reducing one day of handling, by the administration of eCG and PGF at the time of removal of the P4 device (Day 8), results in comparable pregnancy rates than giving eCG on Day 5. A secondary objective was to determine the effect of injectable P4 at the time of device insertion plus EB treatment. Crossbred Bos taurus×Bos indicus beef heifers (n=301) were randomly assigned to 4 treatment groups in a 2 by 2 factorial design. All Heifers received a P4 device (DIB, Syntex, Argentina) plus 2mg EB i.m. (Syntex) at unknown stages of the estrous cycle (Day 0), with or without 50mg of P4 given i.m. at the same time. Heifers were further subdivided to receive PGF (0.150mg d-cloprostenol, Prolise, Tecnopec, Sao Paulo, Brazil) and 400IU of eCG (Novormon, Syntex) i.m. on Days 5 or 8. In all heifers, DIB devices were removed on Day 8 and 1 mg EB was administered i.m. on Day 9. Day 10 was arbitrarily considered as the day of estrus. On Day 17, heifers were bled for plasma P4 concentrations and examined by ultrasonography to determine the number of CL and their diameter. Heifers that had &gt;1 CL or a single CL with diameter ≥18mm received an in vitro-produced (IVP) embryo by nonsurgical transfer performed by the same veterinarian. Pregnancy rates were determined by ultrasonography 30 days later. The effects of Day of eCG administration (Day 5 or Day 8), P4 of treatment (E2 or E2+P4) and the day-by-P4 treatment interaction on the numbers of CL and plasma P4 were analyzed by ANOVA, and the proportion of recipients selected and pregnant were analyzed using non-parametric tests (NPAR1WAY, SAS). There was no significant effect of P4 treatment or the P4-by-day of eCG interaction in any of the parameters evaluated. However, there was a significant effect of day of eCG administration on plasma P4 concentrations (Day 5=2.4±0.3 v. Day 8=1.7±0.2; P=0.03) and the number of CL (Day 5=1.4±0.1 v. Day 8=1.1±0.0; P=0.02) on Day 17. Furthermore, the proportion of recipients pregnant/treated tended (P=0.1) to be higher in heifers in the Day 5 Group (71/151, 47.0%) than in those in the Day 8 Group (61/150, 40.7%). Although delaying the eCG and PGF administration from Day 5 to Day 8 saves one trip through the chute for treatments, it resulted in lower plasma P4 concentrations and tended to decrease pregnancy rates in bovine embryo recipients synchronized with DIB devices and EB and transferred at a fixed time. Furthermore, the administration of injectable P4 at the time of DIB insertion did not affect pregnancy rates.


1988 ◽  
Vol 28 (1) ◽  
pp. 1 ◽  
Author(s):  
RG Holroyd ◽  
PJ Dunster ◽  
PK O'Rourke

The effects of cattle tick infestations on liveweight and fertility of Droughtmaster (1/2 Bos indicus) cows and on calf weaning weights were determined over 3 years. Tick populations on the control (non-dipped) group of cattle fluctuated, with mean annual tick counts/side being 9.5, 8.9 and 13.6 for years 1, 2 and 3 respectively, while the treated group of cattle, which were dipped every 21 days, were free of ticks. Tick counts were not related to fertility or liveweight change in pregnant-lactating cows or to calf growtb or weaning weights. Treatment for ticks significantly (P<0.05) affected liveweight change in pregnant-lactating cows on only a few occasions, and annual liveweight changes were not significantly influenced by treatment. When lactating cow pregnancy rates were low (< 30% for control cows), treatment for ticks increased the pregnancy rate by about 100% in 2 of the 3 years, these differences being significant only in the last year. Calves in the treated group were born significantly earlier in the first year and had significantly lighter birth weights in the third year. Treated calves grew faster to weaning and had higher weaning weights (mean difference 17.9 kg) than control calves but differences were significant in the first and third years only.


2007 ◽  
Vol 19 (1) ◽  
pp. 225 ◽  
Author(s):  
N. Sakagami ◽  
K. Akiyama ◽  
Y. Nakazawa

A precise evaluation of embryo quality is important to estimate the suitability of embryo transfer to recipient animal. Recently, an objective evaluation method was reported for bovine embryos, in which the oxygen consumption of embryos can be noninvasively determined by scanning electrochemical microscopy (SECM) (Shiku et al. 2001 Anal. Chem. 73, 3751–3758). Trimarchi et al. (2000 Biol. Reprod. 62, 1866–1874) suggested that the oxygen consumption reflects the cell number and mitochondrial activity of embryos. The objectives of this study were (1) to examine the oxygen consumption of in vivo-derived embryos by SECM, (2) to investigate the relationship between oxygen consumption and morphological estimation of embryos, and (3) to assess the correlation among the oxygen consumption, embryo viability, and pregnancy rates. Fifty-six embryos were collected from Japanese Black cattle, which were superovulated with a total dose of 20 mg porcine FSH (FSH-R; Kawasaki Pharmaceutical Co., Ltd., Tokyo, Japan) followed by AI. The qualities of collected embryos at the stage of compacted morulae (CM), early blastocysts (EB), and blastocysts (BL) on Day 7 after AI were categorized as grade 1 and grade 2, according to the IETS manual (2002). The oxygen consumption rates of embryos were evaluated by SECM, as previously described by Abe et al. (2004 J. Mamm. Ova Res. 21). Embryos were frozen by programmable freezer in Dulbecco&apos;s PBS containing 1.5 M ethylene glycol, 0.1 M trehalose, and 20&percnt; calf serum. They were thawed by holding the straws in air for 8 s and then immersing them in a 30&deg;C water bath for 15 s. After thawing, the embryos were examined for oxygen consumption. Twenty-eight embryos were then cultured in TCM-199 supplemented with 20&percnt; fetal bovine serum and 0.1 mM &beta;-mercaptoethanol for 24 h to assess the viability of embryos by re-expansion of blastocole. The remaining 28 embryos were transferred to recipients. The pregnancy rates were determined by rectal palpation on Day 70. Data were analyzed by ANOVA. The consumption rates of BL embryos on Day 7 were significantly higher (P &lt; 0.05) than those of CM collected on the same day (0.84 vs. 1.29 &times; 10&minus;14 mol s&minus;1, respectively). A significant difference was also observed in consumption rates between grade 1 and 2 embryos at the BL stage (P &lt; 0.05). After freezing&ndash;thawing, the average oxygen consumption rates of embryos were 0.52 &times; 10&minus;14 mol s&minus;1 for CM (n &equals; 9), 0.67 &times; 10&minus;14 mol s&minus;1 for EB (n &equals; 8), and 0.96 &times; 10&minus;14 mol s&minus;1 for BL (n &equals; 11). The CM embryos with rates of &lt; 0.5 &times; 10&minus;14 mol s&minus;1 and the EB and BL embryos with those &lt; 0.6 &times; 10&minus;14 mol s&minus;1 did not show good morphological appearance after 24 h in culture. Pregnant animals were not obtained from embryos with rates &lt;0.5 &times; 10&minus;14 mol s&minus;1 for CM (n &equals; 5) and &lt;0.7 &times; 10&minus;14 mol s&minus;1 for EB (n &equals; 9). A high pregnancy rate (67&percnt;) was obtained from embryos with rates &gt;1.0 &times; 10&minus;14 mol s&minus;1 for BL (n &equals; 14). These results suggest that the measurement of oxygen consumption of embryos after embryo freezing and prior to embryo transfer may be useful for estimating embryo quality and suitability of embryo transfer.


2007 ◽  
Vol 19 (1) ◽  
pp. 219 ◽  
Author(s):  
S. Moriyasu ◽  
H. Hirayama ◽  
K. Sawai ◽  
S. Kageyama ◽  
S. Aoyagi ◽  
...  

Oxygen consumption is an important indicator of the metabolic activity of living cells, which may provide valuable information for evaluating embryo quality. We have found that the bovine embryos with high oxygen consumption possess stronger potential for further development. However, the relationship between respiratory activity and the pregnancy rate of embryos is still unclear. In this study, we investigated the respiration rates of bisected bovine embryos and the pregnancy rates of demi-embryos after embryo transfer. Compact morula-stage embryos were bisected evenly by micro glass needle. One hundred bisected embryos were incubated for 24 h in embryo culture medium (IVD101; Research Institute for the Functional Peptides, Yamagata, Japan) at 39�C under 5% CO2, 5% O2, 90% N2. After the incubation, demi-embryos were classified into 2 groups: blastocoel-formed (BC) and blastocoel-not-formed (CM) embryos. Oxygen consumption rates of demi-embryos were measured by scanning electrochemical microscopy (SECM; Hokuto Denko Corporation, Tokyo, Japan). Within 3 h after the measurement, 80 demi-embryos were transferred into recipient cows (one demi-embryo/one recipient) at 7–8 days after estrus. Recipient cows were diagnosed for pregnancy by ultrasonography approximately 40 days after estrus. Statistical difference was analyzed by Tukey's post-hoc test and chi-square test. A total of 27 recipient cows became pregnant; the pregnancy rates for cows with CM and BC demi-embryos were 40.6% (13/32) and 29.2% (14/48), respectively. Mean oxygen consumption rates (� 10-14 mol s-1) in pregnant and non-pregnant cows were 0.47 and 0.39 for CM demi-embryos and 0.63 and 0.52 for BC demi-embryos, respectively. Retrospective analysis showed that the respiratory activity of demi-embryos in the pregnant group was higher than those in the non-pregnant group. In particular, the pregnancy rates for demi-embryos with respiratory activity higher than 0.35 in CM and 0.40 in BC groups were 52.0% (13/25) and 35.9% (14/39), respectively. On the other hand, cows with demi-embryos having an oxygen consumption rate under 0.35 in CM (n = 7) and 0.40 in BC (n = 9) groups did not become pregnant. These results demonstrated that bovine demi-embryos with higher respiratory activity showed a high pregnancy rate after embryo transfer. It is generally known that the pregnancy rate after the transfer of bisected embryos is lower than that of whole embryos. The measurement of oxygen consumption by SECM procedures is a useful tool to assess the quality of pre-implantation embryos and may contribute to the improvement of the success rate for bisected embryo transfer.


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