MGA and growth promotants administered to beef feedlot heifers have no effect on subsequent oocyte quality or in vitro embryo production

We previously reported that follicular wave synchronization, by removal of the dominant follicle on Day 5 after ovum pickup (OPU), was effective in increasing oocyte quality in the developing follicles (Imai et al. 2006 32th Annual Conference of the IETS, poster presentation no. 277). The current study was designed to examine the effect of superstimulatory treatment to induce subsequent follicular wave synchronization on embryo production by OPU and IVM-IVF-IVC in Holstein dry cows. Cows were reared under the same feeding and environmental conditions, and 2 OPU sessions were conducted in each cow. In the first session, OPU was performed in 8 cows on arbitrary days of the estrous cycle by using a 7.5-MHz linear transducer with needle (Cova needle, Misawa Medical, Tokyo, Japan) connected to an ultrasound scanner (SSD-1200, Aloka, Tokyo, Japan). Follicles larger than 8 mm in diameter were then aspirated and a CIDR was inserted on Day 5 (the day of first OPU session = Day 0). Cows then received 30 mg of FSH (Antrin-R10; Kawasaki Mitaka Pharmaceutical Co., Tokyo, Japan) twice a day from Days 7 to 10 in decreasing doses (6, 6, 4, 4, 3, 3, 2, 2 mg) by i.m. injection. Cloprostenol (PGF; Clopromate C; Sumitomo Pharmaceuticals Co., Tokyo, Japan; 0.75 mg) was administered in the morning of Day 9 (third day of superstimulation). The second OPU session was performed 48 h after PGF administration (Day 11), and only follicles larger than 5 mm in diameter were aspirated. The CIDR was removed from the cows just before OPU. Collected oocytes were evaluated by their cumulus cell morphology, cytoplasmic color, and density. Grades 1 and 2 COC were matured, fertilized, and cultured as described by Imai et al. [2006 J. Reprod. Dev. 52(Suppl.), S19–S29]. Embryo development was assessed by the cleavage rate on Day 2 and by the blastocyst formation rate on Days 7 to 8 (the day of insemination = Day 0). Data were analyzed by Student's t-test. There were no differences in the mean (� SD) number of aspirated follicles or collected oocytes between the first (32.5 � 6.8 and 26.0 � 12.7, respectively) and second (29.3 � 10.4 and 19.0 � 9.4, respectively) OPU sessions (P > 0.1). The percentage of Grade 1 and 2 oocytes for the second OPU session (90.5 � 13.8%) was significantly higher (P < 0.01) than for the first OPU session (63.1 � 6.3%), and significant differences were found for cleavage (79.4 � 14.1, 61.8 � 25.1, P < 0.01) and blastocyst rates (68.1 � 16.7, 24.2 � 22.3, P < 0.001) between sessions. The mean numbers of blastocysts obtained per session were 4.3 � 2.9 and 12.8 � 8.7 in the first and second sessions, respectively (P < 0.01). These results indicate that superstimulatory treatment and subsequent follicular wave synchronization were effective on in vitro embryo production by increasing the oocyte quality.

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Effects of a high-energy diet on oocyte quality and in vitro embryo production in Bos indicus and Bos taurus cows

Journal of Dairy Science ◽

10.3168/jds.2014-8858 ◽

2015 ◽

Vol 98 (5) ◽

pp. 3086-3099 ◽

Cited By ~ 24

Author(s):

J.N.S. Sales ◽

L.T. Iguma ◽

R.I.T.P. Batista ◽

C.C.R. Quintão ◽

M.A.S. Gama ◽

...

Keyword(s):

Bos Taurus ◽

Bos Indicus ◽

Oocyte Quality ◽

High Energy ◽

Embryo Production ◽

In Vitro Embryo Production

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Beneficial Effects of Melatonin in the Ovarian Transport Medium on In Vitro Embryo Production of Iberian Red Deer (Cervus elaphus hispanicus)

Animals ◽

10.3390/ani10050763 ◽

2020 ◽

Vol 10 (5) ◽

pp. 763 ◽

Cited By ~ 2

Author(s):

Irene Sánchez-Ajofrín ◽

María Iniesta-Cuerda ◽

Patricia Peris-Frau ◽

Alicia Martín-Maestro ◽

Daniela-Alejandra Medina-Chávez ◽

...

Keyword(s):

Red Deer ◽

Oocyte Quality ◽

Scale Production ◽

Post Mortem ◽

Embryo Production ◽

Transport Medium ◽

Iberian Red Deer ◽

Blastocyst Quality ◽

In Vitro Embryo Production

A major limiting factor for the development of in vitro embryo production (IVP) in wild species, such as Iberian red deer, compared to livestock animals is the poor availability and limited access to biological material. Thus, the use of post-mortem ovaries from slaughtered animals represent a source of oocytes for the large scale production of embryos needed for research and to improve the efficiency of IVP. However, these oocytes are not as developmentally competent as their in vivo counterparts. Moreover, oocytes are usually obtained from ovaries that have been transported for long distances, which may also affect their quality. In order to overcome the issues associated with prolonged storage times of post-mortem material, in this study we examined the effect of melatonin supplementation to the ovary transport medium on oocyte quality, embryo yield, and blastocyst quality in Iberian red deer. When necessary, sheep was used as an experimental model due to the large number of samples required for analysis of oocyte quality parameters. Oocytes were in vitro matured and assessed for early apoptosis; DNA fragmentation; reactive oxygen species (ROS); reduced glutathione (GSH) content, mitochondrial membrane potential, and distribution; and relative abundance of mRNA transcript levels. After in vitro fertilization, embryo rates and blastocyst quality were also investigated. The results revealed that melatonin treatment significantly increased intracellular level of GSH in sheep oocytes. Moreover, the percentage of cleavage and blastocyst yield in red deer was greater compared to the Control group and there was lower abundance of oxidative stress- and apoptosis-related SHC1, TP53, and AKR1B1 mRNA transcripts in blastocysts for the Melatonin group. In conclusion, the supplementation of melatonin to the ovary storage medium had a positive effect on the developmental competence and quality of resulting blastocysts in Iberian red deer.

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205 GLUCOSE-6-PHOSPHATE DEHYDROGENAZE ACTIVITY IN OVINE OOCYTES IS ASSOCIATED WITH PRE-IMPLANTATION EMBRYONIC DEVELOPMENT IN VITRO

Reproduction Fertility and Development ◽

10.1071/rdv23n1ab205 ◽

2011 ◽

Vol 23 (1) ◽

pp. 201 ◽

Cited By ~ 1

Author(s):

F. Asghari ◽

M. Shahidi ◽

Y. Chashnidel ◽

H. Deldar ◽

Z. Ansari-Pirsaraei ◽

...

Keyword(s):

Oocyte Quality ◽

G6pd Activity ◽

Blue Staining ◽

Control Group ◽

Embryo Production ◽

Brilliant Cresyl Blue ◽

Cresyl Blue ◽

In Vitro Embryo Production ◽

Humidified Air

A large proportion of ovine oocytes fail to develop into viable embryos following maturation, fertilization, and culture in vitro. Accurate, fast, and noninvasive predictors of ovine oocyte quality are therefore in urgent need for oocyte selection before in vitro maturation (IVM). Recent studies have shown that oocyte competence can be predicted through the presence of the glucose-6-phosphate dehydrogenase (G6PD) enzyme, as indicated by brilliant cresyl blue (BCB), a dye that can be degraded by G6PD. Thus, oocytes that have completed their growth phase show decreased G6PD activity and exhibit cytoplasm with a blue colouration (BCB+), whereas growing oocytes are expected to have a high level of G6PD, which results in colourless cytoplasm (BCB–). The brilliant cresyl blue staining test, as a noninvasive intrinsic criterion, has been successfully used to identify the more competent oocytes in various species. Therefore, this study aimed to investigate whether BCB staining, as an indicator of G6PD activity, can be used to select developmentally competent ovine oocytes before IVM and thereby increase the efficiency of in vitro embryo production. Ovine ovaries were obtained from a local slaughterhouse and transported to the laboratory, where cumulus–oocyte complexes (COC) were recovered by slicing the ovaries. Only oocytes with one or more complete layers of unexpanded cumulus cells and a homogeneous cytoplasm were used. The COC were exposed to 26 mM BCB diluted in modified Dulbecco’s PBS for 90 min at 39°C in humidified air. After BCB exposure, the COC were examined under a stereomicroscope and divided into 2 groups: BCB+ (blue cytoplasm, low G6PD activity) and BCB– (colourless cytoplasm, high G6PD activity). Cumulus–oocyte complexes in the control group were incubated for IVM directly after selection, without exposure to BCB dye. After IVM, oocytes were subjected to IVF followed by embryo culture for 7 days (5% CO2, 39°C, humidified air). Results were analysed by a chi-square test, and P < 0.05 was considered statistically significant. The proportion of oocytes that cleaved by Day 2 after insemination was significantly (P < 0.05) higher for the control and BCB+ groups [67.3% (68/101) and 71.7% (81/113), respectively] than for the BCB– group [50.5% (46/91)]. Significant differences among groups were also observed on Day 7 after fertilization, when the embryos reached the blastocyst stage of development. The BCB+ group yielded a significantly (P < 0.05) higher proportion of blastocysts [34.5% (39/113)] than both the control [20.8% (21/101)] and BCB– [4.3% (4/93)] groups. In addition, the blastocyst rate of development in the control group was significantly (P < 0.05) higher than that for the BCB– group. In conclusion, results of this study show that selection of ovine oocytes based on G6PD activity through the BCB test can be used as an efficient predictor of in vitro embryonic developmental competence. This positive predictive parameter of oocyte quality may also be useful in increasing the efficiency of blastocyst production during in vitro embryo production procedures in the ovine.

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245 EFFECTS OF Gyr (BOS TAURUS INDICUS) DONOR MITOCHONDRIAL DNA AND OVUM PICKUP ORDER ON IN VITRO EMBRYO PRODUCTION

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab245 ◽

2015 ◽

Vol 27 (1) ◽

pp. 212

Author(s):

T. L. C. Pinto ◽

B. C. Lopes ◽

M. B. D. Ferreira ◽

T. M. Goncalves ◽

J. C. Souza ◽

...

Keyword(s):

Mixed Model ◽

Bos Taurus ◽

Bos Indicus ◽

Covariance Structure ◽

Oocyte Quality ◽

Embryo Production ◽

Bos Taurus Indicus ◽

Hindiii Restriction Site ◽

In Vitro Embryo Production

Ovum pick (OPU) up is a technique upon which in vitro embryo production (IVP) depends. It permits donor cow maternal ancestry to be assessed by mtDNA analysis. Repetitive ablation of follicles is thought to interfere with the donor cow endocrine profile and influence embryo yield. The objective was to evaluate the effects of mtDNA and OPU order on IVP fertility traits. Gyr donors (85) were submitted to 363 OPU sessions (5 OPU sessions/donor). Donor mtDNA was extracted from leukocytes and sorted by the presence of the HindIII restriction site within the amplified region, indicative of Bos taurus taurus mtDNA (Paneto et al. 2008 Genet. Mol. Res. 7, 592). All animals in the donor pedigree were classified, and the population was divided into two groups according to their maternal genetic grouping: Bos taurus indicus and Bos taurus taurus. For statistical analyses, data from 5 OPU sessions per each donor were submitted to the mixed model procedure of SAS® (SAS Institute Inc., Cary, NC, USA), using the lowest Akaike value to determine the best covariance structure between repeated OPU session results. The model included the effect of OPU session order (1–5) and mtDNA-based maternal grouping (Bos taurus v. Bos indicus) as independent variables. Total and viable oocytes, as well as blastocyst yield per OPU session, were the dependent variables studied. The means of total and viable oocytes and blastocysts produced per donor per OPU were compared by the Tukey test, at 5% significance. The combined OPU sessions resulted in 6084 oocytes, 2537 embryos, which produced 1105 pregnancies. Mean numbers (n = 42 donors/OPU session) of total and viable oocytes between OPU sessions 1 (31.9 ± 4.6 and 19.2 ± 2.9), 2 (35.3 ± 3.5 and 21.2 ± 2.2), 3 (28.9 ± 3.7 and 19.3 ± 2.4), 4 (25.0 ± 5.2 and 18.9 ± 33), and 5 (20.5 ± 6.3 and 13.8 ± 4.0) did not differ. Mean blastocyst production after IVP between OPU sessions 1 (6.5 ± 1.4), 2 (5.15 ± 1.04), 3 (5.5 ± 1.1), 4 (5.8 ± 1.6), and 5 (5.2 ± 1.9) was similar. Mean viable oocyte number was greater (P < 0.0001) for B. t. taurus (21.7 ± 1.4, n = 192 OPU sessions) compared with B. t. indicus (16.1 ± 0.9, n = 138 OPU sessions) maternal genetic groupings. However, in vitro blastocyst yields were similar (P = 0.23) between maternal genetic groupings (7.3 ± 1.6 and 6.2 ± 1.4 for B. taurus and B. indicus, respectively). In conclusion, repeated OPU sessions did not reduce oocyte and embryo yields as expected. Maternal B. taurus genetic origin was associated with higher oocyte quality, although it was not translated into higher embryo yields after in vitro culture. Results warrant further research, which may result in additional selection criteria for OPU Gyr donors considering their maternal genetic background.

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Effects of vulvar width and antral follicle count on oocyte quality, in vitro embryo production and pregnancy rate in Bos taurus taurus and Bos taurus indicus cows

Animal Reproduction Science ◽

10.1016/j.anireprosci.2020.106357 ◽

2020 ◽

Vol 217 ◽

pp. 106357

Author(s):

Gisvani Lopes de Vasconcelos ◽

Ellen Vasconcelos da Cunha ◽

Renata Maculan ◽

Jesús Alfonso Sánchez Viafara ◽

Anderson Weiny Barbalho Silva ◽

...

Keyword(s):

Pregnancy Rate ◽

Bos Taurus ◽

Antral Follicle ◽

Oocyte Quality ◽

Antral Follicle Count ◽

Embryo Production ◽

Bos Taurus Indicus ◽

In Vitro Embryo Production

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Laparoscopic Ovum Pick-Up Followed by In Vitro Embryo Production and Transfer in Assisted Breeding Programs for Ruminants

Animals ◽

10.3390/ani11010216 ◽

2021 ◽

Vol 11 (1) ◽

pp. 216

Author(s):

Hernan Baldassarre

Keyword(s):

Pregnancy Rate ◽

Commercial Application ◽

Embryo Production ◽

Breeding Programs ◽

Sheep And Goats ◽

Embryo Recovery ◽

Marker Selection ◽

Holstein Calves ◽

In Vitro Embryo Production

The potential of laparoscopic ovum pick-up (LOPU) followed by in vitro embryo production (IVEP) as a tool for accelerated genetic programs in ruminants is reviewed in this article. In sheep and goats, the LOPU-IVEP platform offers the possibility of producing more offspring from elite females, as the procedure is minimally invasive and can be repeated more times and more frequently in the same animals compared with conventional surgical embryo recovery. On average, ~10 and ~14 viable oocytes are recovered by LOPU from sheep and goats, respectively, which results in 3–5 transferable embryos and >50% pregnancy rate after transfer. LOPU-IVEP has also been applied to prepubertal ruminants of 2–6 months of age, including bovine and buffalo calves. In dairy cattle, the technology has gained momentum in the past few years stemming from the development of genetic marker selection that has allowed predicting the production phenotype of dairy females from shortly after birth. In Holstein calves, we obtained an average of ~22 viable oocytes and ~20% transferable blastocyst rate, followed by >50% pregnancy rate after transfer, declaring the platform ready for commercial application. The present and future of this technology are discussed with a focus on improvements and research needed.

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