189 PREGNANCY RATES IN HEAT-STRESSED DAIRY CATTLE RECEIVING ONE OR TWO IN VITRO-PRODUCED EMBRYOS IN A TIMED EMBRYO TRANSFER PROGRAM

2006 ◽  
Vol 18 (2) ◽  
pp. 202
Author(s):  
M. Franco ◽  
J. Block ◽  
F. D. Jousan ◽  
L. A. de Castro e Paula ◽  
A. M. Brad ◽  
...  

The objective was to determine whether transfer of two embryos would increase pregnancy rates in heat-stressed dairy recipients receiving an in vitro-produced embryo transferred into the uterine horn ipsilateral to the corpus luteum (CL). Such a treatment would increase the likelihood that the cow receives at least one embryo competent for sustained development. In addition, transfer of two embryos into the ipsilateral uterine horn is likely to increase the amounts of interferon-tau and other embryonic-signaling molecules in the uterus. A total of 32 virgin crossbreed heifers and 26 lactating crossbreed cows were used as timed embryo transfer recipients after being subjected to an ovulation synchronization protocol as follows: GnRH (100 �g) and insertion of previously used progesterone-containing CIDR on Day -10, prostaglandin F2� and CIDR removal on Day -3, and GnRH (100 �g) on Day 0 (day of anticipated ovulation). All recipients had a palpable CL on Day 6 and were randomly selected to receive one (n = 31 recipients) or two (n = 27) embryos on Day 7. At Day 64, the pregnancy rate tended to be higher (P = 0.07) for cows than for heifers. While not significant, heifers that received two embryos tended to have lower pregnancy rates than those that received a single embryo (20% vs. 41%); there was no difference in pregnancy rate in cows (50% for two embryos vs. 57% for one embryo). Pregnancy losses between Day 64 and Day 127 occurred in one group only cows receiving two embryos. In that group, pregnancy rate was 50% at Day 64 but 17% at Day 127. Overall, there was no difference in pregnancy rates at day 127 between cows and heifers, but recipients that received two embryos (17% for cows and 20% for heifers) had lower pregnancy rates (P < 0.03) than recipients that received one embryo (57% for cows and 41% for heifers). Only one animal, a cow, had twin fetuses at day 127. In conclusion, unilateral transfer of two embryos failed to improve pregnancy rates of dairy cattle exposed to heat stress. The fact that fetal loss occurred sooner for heifers than for cows points out the importance of uterine capacity as a limiting factor for maintenance of fetal development of two conceptuses. The suitability of timed embryo transfer was evident from the high pregnancy rates achieved with crossbreed females that received a single embryo. This work was supported by BARD Grant No. US-1551-14, USDA TSTAR Grant No. 2004-14135-14715, Grant No. 2001-12101-11318 from the USDA-IFAFS Program, and CAPES Grant No. 134202-1).

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&lt;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&lt;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


2015 ◽  
Vol 27 (1) ◽  
pp. 164
Author(s):  
R. C. Fry ◽  
K. L. Fry ◽  
H. A. McCartney ◽  
W. R. Geddes ◽  
K. Geddes

The aim of this experiment was to investigate the effect of day of synchrony on the pregnancy rate of recipients following the transfer of Day 7 IVF embryos. In addition, the effect of IVF embryo grade and corpus luteum (CL) grade of recipients was determined. A total of 317 cumulus-oocyte complexes collected from 24 dry Brahman cows by TVR were matured, fertilized, and cultured under standard in vitro production procedures (Fry et al. 2003 Theriogenology 59, 446). A total of 89 (44 Grade 1, 43 Grade 2, and 2 Grade 3, IETS classification) in vitro-produced embryos were transferred to parous 4- to 9-year-old dry Brahman cross recipient cattle 7 days after IVF. Two groups of recipient cows were synchronised one day apart with an 8-day CIDR/pg protocol so that oestrous would be concentrated over 3 days with the middle day aligning with the day of IVF (Day 0). Donors that produced a large number of IVF embryos had these divided and transferred into recipients either on Day –1 or Day +1 of synchrony, and those producing less than 4 IVF embryos were transferred into recipients on Day 0. At embryo transfer the ovaries of the recipient were palpated and then scanned by rectal ultrasound and the grade of CL noted (Grade 1 = large distinct CL by palpation, Grade 2 = small distinct CL by palpation, Grade 3 = CL not distinguishable by palpation). Pregnancy was diagnosed by ultrasound scanning on Day 92. Although recipient numbers were low, differences in pregnancy rate between groups were analysed by Chi-squared. Data from the 2 Grade 3 embryos transferred were not included in the analysis (0/2 pregnant). Similar (P > 0.05) pregnancy rates were found when Day 7 IVF embryos were transferred to either Day 6 (17/32 = 53%), Day 7 (9/24 = 38%), or Day 8 (14/31 = 45%) recipients. Furthermore, neither the grade of the embryo (Grade 1: 20/44 = 45%, Grade 2: 20/43 = 47%) nor the grade of recipient CL (Grade 1: 17/45 = 38%, Grade 2: 17/29 = 59%, Grade 3: 6/13 = 46%) effected pregnancy rate (P > 0.05). This experiment demonstrates the flexibility of the IVF embryo to achieve an acceptable pregnancy rate over a range of recipient stages thereby allowing a high usage rate of good-quality recipients in an IVF embryo transfer program.


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.


2010 ◽  
Vol 22 (1) ◽  
pp. 301
Author(s):  
B. G. Moura ◽  
J. Almeida ◽  
F. L. Lima ◽  
G. Balbi ◽  
R. Calmerani ◽  
...  

The aim of the work was to study the effects of year period, technical team, breed, beef cattle and dairy cattle on the pregnancy rates in fresh embryos used in bovine transfer of IVF programs. The study was carried out at the fertilization laboratory In Vitro Nyltta Britto de Carvalho, in partnership with In Vitro Brazil, located at the Boa Vista farm, Barra do Pirai, during August 2007 to September 2008, seeking subsidies to improve the use of the technique in the field. During that period, aspirations and inovulations in 3 different periods I (August to December), II (January to April), and III (May to September) were carried out. The jobs were accomplished by 9 technical teams (A, B, C, D, E, F, G, H, and I) rendering services to the laboratory, by working with 2 beef breeds (Brahman and Nelore) and 3 dairy breeds (Gir, Girolando, and Holstein). The different breed receivers were synchronized, and in general, from 6 to 8 days after heat, they received embryo transfer, the cervical way, under low epidural anesthesia, where each female received 1 fresh embryo of IVF. All cows were submitted to gestation diagnosis by rectal palpation and ultrasonography, in general, 42 days after embryo transfer. The numbers of embryo transferred and pregnancy rates were submitted to the chi-square test, which presented significant differences (P < 0.05). There were pregnancy rates of 36.25%a (n = 960), 39.83%a (n = 1180), and 32.59%b (n = 919) in the I, II, and III periods, respectively. Among the 9 technical teams, there were verified pregnancy rates (%) of 33.51d (n = 1313), 30.30d (n = 330), 35.00cd (n = 405), 39.24cd (n = 1060), 59.25a (n = 7), 33.33d (n = 24), 53.57bc (n = 28), 43.31c (n = 157), and 58.33ab (n = 12) for A, B, C, D, E, F, G, H, and I teams, respectively. Among breeds there were rates (%) of 36.89ab (n = 412), 34.68b (n = 1286), 35.13ab (n = 74), 38.94a (n = 1140), and 37.80ab (n = 82) for Brahman, Nelore, Gir, Girolando, and Holstein, respectively. In the study, pregnancy rates (%) of 35.21b (n = 1698) in beef cattle and 38.65a (n = 1296) in dairy cattle were observed. The differences in pregnancy rates with respect to the evaluated factors, may be explained by individual, breed, and nutritional variations of the animals. There are few data in the literature with results on the embryo transfer use of IVF bovine under field conditions.


2018 ◽  
Vol 30 (1) ◽  
pp. 177
Author(s):  
Y. Hashiyada ◽  
Y. Aikawa ◽  
H. Matsuda ◽  
T. Yamanouchi ◽  
Y. Goto ◽  
...  

Monozygotic twin bovine embryos can be produced by blastomere separation of 2-cell embryos and commercial well-of-the-well (WOW) culture dish (Hashiyada et al. 2016 Reprod. Fertil. Dev. 28, 178) obtaining 60% and 48% of blastocyst formation and monozygotic blastocyst pairs, respectively. The present study was conducted to evaluate the fertility of blastocysts derived from this production system in Japanese Black beef cattle. Embryos were produced using oocytes collected by ovum pick-up technique. TCM-199 supplemented with 5% calf serum (CS), Brackett-Oliphant solution supplemented with 10 mg mL−1 BSA, and CR1aa containing 5% CS, were used for each culture step: in vitro maturation, fertilization, and culture (IVM,IVF, and IVC). Two-cell stage embryos were obtained 24 to 27 h post-insemination. Zonae pellucidae were removed by exposure to 0.25% pronase. Then, embryos were separated into blastomeres by gentle pipetting in IVC medium. Each blastomere was introduced into a single conical microwell of 25 wells, each having a diameter and depth of ~287 μm and 168 μm (Dai Nippon Printing, Tokyo, Japan). Blastomeres in wells were cultured covered with a droplet of 2.5 μL of IVC medium/well. The developed blastocysts in pairs on 7 days post-insemination were used for transfer. Single embryos of monozygotic twin embryos were transferred to Japanese Black cattle with a generally small body frame to produce twin calves from a set of recipients. Twin embryos were transferred in pairs to unilateral of uterus of non-lactating Holstein cows. Pregnancy and twin pregnancy were determined at 30 days of gestation by ultrasonography and were reconfirmed at 60 days with detection of fetal loss. Statistical significance was analysed by Fisher’s exact test. There was no significant difference in pregnancy rate or twin pregnancy rate between single embryo transfer (7/14, 50% and 2/7, 28.6%) and twin embryo transfer (9/21, 42.9% and 4/21, 19%). In either transfer method, fetal loss was not observed in diagnosis carried out at 60 days by ultrasonography. To date, 2 pairs of twin calves have been obtained from twin pregnant cows by twin embryo transfer within the normal range of gestation length (286 and 288 days) and birth weight (31-40 kg). These results indicate that blastocysts developed from blastomeres separated from 2-cell embryos by culturing with commercial WOW culture dish had fertility similar to that of intact embryos derived from standard in vitro culture and further demonstrate the possibility of production of normal twin calves.


2006 ◽  
Vol 24 (1) ◽  
pp. 23-28 ◽  
Author(s):  
David Johnson

Over a period of three years, acupuncture was offered to patients entering assisted reproduction therapy. Acupuncture sessions were given at varying, but usually weekly, intervals during the in vitro fertilisation (IVF) cycle, and immediately before and after embryo transfer. Twenty two patients (average age 36.2 years) were treated over a total of 26 IVF cycles and 15 pregnancies were achieved, as determined by presence of foetal heartbeat on ultrasound at four weeks post embryo transfer. This was a success rate of 57.7% compared with 45.3% for patients in the IVF unit not treated with acupuncture (P>0.05). Relaxing effects were noted following acupuncture and it is speculated that this may have contributed to the increase in pregnancy rate for the acupuncture group.


2006 ◽  
Vol 18 (2) ◽  
pp. 202 ◽  
Author(s):  
O. Dochi ◽  
M. Tanisawa ◽  
S. Goda ◽  
H. Koyama

Repeat-breeding is one of the important factors that affect dairy management. The objective of this study was to investigate the effect of transfer of frozen–thawed IVF embryos on pregnancy in repeat-breeder Holstein cattle. Cumulus–oocyte complexes (COCs) were collected by aspiration of 2–1-mm follicles from ovaries obtained at a local abattoir. COCs were matured for 20 h in TCM-199 supplemented with 5% calf serum (CS) and 0.02 mg/mL of FSH at 38.5°C under a 5% CO2 atmosphere in air. Matured oocytes were inseminated with spermatozoa of 5 × 106/mL in BO solution (Brackett and Oliphant 1975 Biol. Reprod. 12, 260–274) containing 10 mM hypotaurine and 4 units/mL heparin. After 18 h of gamete co-culture, presumptive zygotes were cultured in CR1aa (Rosenkrans et al. 1991 Theriogenology 35, 266) supplemented with 5% CS for 8 days at 38.5°C under 5% CO2, 5% O2, 90% N2 atmosphere in air. After in vitro fertilization, Day 7 and Day 8 blastocysts were frozen in 1.5 M ethylene glycol (EG) in Dulbecco's PBS (DPBS) supplemented with 0.1 M sucrose and 20% CS. Embryos were transferred into a freezing medium, loaded into 0.25-mL straws, and allowed to stand for 15–20 min for equilibration. The straws were then plunged into a −7°C methanol bath of a programmable freezer for 1 min, seeded at −7°C, maintained at −7°C for 15 min, cooled to −30°C at the rate of −0.3°C/min, and then plunged into liquid nitrogen. Recipient animals (43 heifers, 131 cows) included those that did not conceive after being artificially inseminated (AI) 3 to 15 times. The frozen–thawed IVF embryos were directly transferred to the recipient animals 7 days after estrus or AI. Pregnancy rates were analyzed by chi-square test. The results are presented in Table 1. There were no significant differences in the pregnancy rates between treatments. However, a slightly higher pregnancy rate was achieved by embryo transfer after AI. These results suggest that embryo transfer may increase the pregnancy rate in repeat-breeder Holstein cattle. Table 1. Pregnancy rates after transfer of IVF frozen–thawed embryos in repeat-breeder Holstein cattle


2008 ◽  
Vol 20 (1) ◽  
pp. 155
Author(s):  
M. Urakawa ◽  
A. Ideta ◽  
K. Hayama ◽  
K. Tsuchiya ◽  
Y. Aoyagi

Repeat-breeding Holstein cows (RBCs), i.e. cows that have undergone three or more AIs, have reduced pregnancy rates. The present study aimed to determine whether application of a combination of a PRID� (ASKA Pharmaceutical Co., Inc., Tokyo, Japan) + prostaglandin F2α (PGF2α) for estrus synchronization and AI + embryo transfer (ET) would improve pregnancy rates in RBCs. All RBCs selected showed a normal interval between the previous insemination and estrus, and were free of clinical disorders. The RBCs (n = 107) received a PRID for 9 days with a capsule containing 10 mg of estradiol benzoate, 0.5 mg IM of cloprostenol (PGF2α, Resipron�-C; ASKA, Japan) at 2 days before PRID removal. The cows were inseminated with frozen–thawed semen from a bull at 0 to 12 h after onset of estrus. Seven days after AI, single in vitro-produced embryos were transferred to the same RBCs. Pregnancy was diagnosed by palpation per rectum at e60 days post-insemination. Exp. 1 estimated the response to estrus synchronization treatment (n = 107). The factors examined were lactation number (mean � SD: 2.3 � 1.5, range: 1–6), insemination number (5.2 � 2.2, range: 3–13), and the interval from calving to next AI (275.9 � 97.6 days, range: 93–650 days). Exp. 2 compared the pregnancy rate in control RBCs (n = 161) and PRID-treated RBCs (n = 100). The factors examined included lactation number (2.5 � 1.6, range: 1–12), insemination number (6.0 � 2.5, range: 3–19), the interval from calving to next AI (288.0 � 102.1 days, range: 102–797 days), and whether the ET was into the uterine horn ipsilateral or contralateral to the CL in a total of 261 RBCs. Data were analyzed using logistic regression. The results for Exp. 1: 103 cows (96.3%) were confirmed to be in estrus (2.0 � 0.3 days after PRID removal). Although the frequency of estrus was affected by the lactation number (P = 0.014) and the interval from calving to the first AI (P = 0.005), the AI number did not have an affect (P = 0.624). Exp. 2: PRID synchronization had no effect on pregnancy rate (control: 37.3%, 60/161; and PRID-treated: 40.0%, 40/100). For the RBCs, the pregnancy rate was not affected by lactation number (P = 0.093), AI number (P = 0.694), or the interval from calving to next AI (P = 0.762). No significant difference (P = 0.428) in pregnancy rate was observed in cows when the embryos were transferred into the uterine horn contralateral to the CL (43.5%, 20/46), compared to those embryos transferred into the uterus horn ipsilateral to the CL (37.2%, 80/215). In conclusion, our findings suggest that the pregnancy rate may be improved by considering lactation number and the interval from calving to the next AI of RBCs in estrus synchronization using a combination of PRID + PGF2α and AI + ET.


2007 ◽  
Vol 19 (1) ◽  
pp. 297
Author(s):  
S. Li ◽  
W. Yu ◽  
J. Fu ◽  
Y. Bai ◽  
F. Jin ◽  
...  

Data collected from commercial embryo transfer programs in 63 farms in China during June 2002 to December 2005 was analyzed to examine the effects of various factors (biopsy, freezing, sample size, embryo development and quality, in vitro culture, and recipient quality) on pregnancy rates of in vivo-biopsied embryos. Embryos were flushed from superovulated dairy cattle and subjected to a biopsy for sexing determination using protocols and sexing kits supplied by AB Technology Ltd. Fresh embryos were implanted on the same day or frozen with AG freeze medium (AB Technology Ltd., Pullman, WA, USA) for later transfer. Recipients were synchronized with CIDA + PG protocols. Embryos were cultured in 6-well dishes containing 1.3 mL of holding medium (AB Technology Ltd.) in each well at room temperature (20–25�C) for examination of embryo survival in vitro. The chi-square test was used in statistic analysis. The implantation of fresh embryos after biopsy did not affect pregnancy rates (49.6%, 257/518) compared to that of non-biopsied fresh and frozen–thawed embryo groups (52.9%, 47/140 and 46.6%, 177/380, respectively). However, for biopsied embryos subjected to frozen and thawed procedures before implantation, particularly for those subjected to the removal of a larger biopsy, a reduced pregnancy rate was observed (41.8%, 297/710; P &lt; 0.01). Pregnancy rates among biopsied embryos at 3 different development stages (morula-early blastocyst, blastocyst, and expanded blastocyst) were not different. Similar results were found between embryo groups of grade 1 and 2. A significant decrease in pregnancy rate (0/10) was observed with embryos held in vitro for a longer period of time (&gt;5 h), suggesting detrimental effects of in vitro conditions on embryo survival. The highest pregnancy rate (68.0%) was observed in recipients synchronized for the first time before being implanted with biopsied embryos. Significant decreases in such rates were found in recipients synchronized for the second or third times or those with an abortion history at the first or second synchronization-implantation treatment (P &lt; 0.01). Better pregnancy rates (45.6%, 41/90; 46.1%, 76/165; and 45.5%, 5/11) were obtained for recipients implanted with biopsied embryos at Days 7.5, 8.0, and 8.5 post-heat detection, respectively, compared to 16% at Day 7 (3/18, P &lt; 0.05). It is concluded that mechanical treatment (cutting) does not reduce the survival of biopsied embryos; however, cryopreservation reduces their ability to survive in vivo. The analyses also suggest that holding embryos in vitro should not be longer than 5 h unless more favorable in vitro conditions can be provided. To achieve better results of implantation of biopsied embryos, embryo transfer should be performed during 7.5–8.5 days post-estrus, and the healthy recipients synchronized for the first time should be used.


2019 ◽  
Vol 31 (1) ◽  
pp. 181
Author(s):  
G. Gamarra Lazo ◽  
D. Di Scala ◽  
S. Maunas ◽  
R. Chaubet ◽  
S. Lacaze

We previously demonstrated the success of in vitro embryo production (IVP) in Lidia breed cattle (Gamarra Lazo et al. 2017 Reprod. Fertil. Dev. 30, 187). As in other species, the success of IVP is linked to the birth of calves from this technique. In the Lidia breed, an important factor to consider is the use of Lidia recipients in order to keep the temperament characteristic of this breed to next generations. The aim of the study was to produce ovum pickup (OPU)-IVP calves in the Lidia breed and to assess the effects of recipient and embryo related factors (status of the recipients; development stage of IVF embryos) on pregnancy rate following embryo transfer. Ovum pickup-IVP embryos from Lidia breeds were produced by a standard protocol (Gamarra Lazo et al. 2017 Reprod. Fertil. Dev. 30, 187). Numbers of blastocysts and expanded blastocysts were recorded on Day 7. A total of 27 blastocysts (B) and 34 expanded blastocysts (EB) of excellent quality (grade 1 according to IETS classification) were selected for fresh transfer. All embryos were transferred to Lidia breed recipients (heifers or cows) by a single operator under similar environmental and field conditions. Recipients were synchronized by subcutaneous insertion of an ear implant of 3.3mg of Norgestomet (Crestar®, MSD, Courbevoie, France) for 9 days. Two days before implant withdrawal, 0.5mg of Cloprostenol (Estrumate®, MSD) was injected. No oestrous detection was performed and synchronized females were selected as recipients when they presented a well developed corpus luteum at Day 9 after implant withdrawal (Day 6 to 7 after the expected oestrus). Blood samples were collected from recipients to determine pregnancy status using the bovine pregnancy associated glycoprotein (Idexx, Westbrook, ME, USA) 50-60 days after transfer. Pregnancy rates were analysed by chi-square analysis to compare results between heifers and cows and between B and EB embryo stages. The overall pregnancy rate after transfer of IVP fresh embryos from Lidia breed averaged 41.0% (n=25). A higher pregnancy rate was achieved in cows compared to heifers [51.2% (21/41) v. 20.0% (4/20) respectively, P&lt;0.05]. There was no difference in pregnancy rate between grade 1B [37% (10/27)] and EB [44.1% (15/34)] embryos (P&gt;0.05). Surprisingly, these results suggest that Lidia breed cows are the best recipients for OPU-IVP embryos. This may be related to the limited feasibility of manipulating the uterine horn during the embryo transfer in Lidia breed heifers, which have a low weight (less than 280kg) and present a narrow rectum diameter. It has been also observed that the cervix is very thin and difficult to cross, thus increasing the stress and potentially inflammatory and immune products secretion. Development stage of embryos did not affect pregnancy rate. To our knowledge, no OPU-IVP Lidia breed calves have been reported previously following transfer into Lidia breed recipients. In the current work, 13 OPU-IVP Lidia breed calves were born. Therefore, we confirmed the possibility of applying OPU-IVP and embryo transfer techniques in this breed within a genetic program.


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