Dietary propylene glycol and in vitro embryo production after ovum pick-up in heifers with different anti-Müllerian hormone profiles

2015 ◽  
Vol 27 (8) ◽  
pp. 1249 ◽  
Author(s):  
G. Gamarra ◽  
C. Ponsart ◽  
S. Lacaze ◽  
B. Le Guienne ◽  
P. Humblot ◽  
...  
Keyword(s):  
Propylene Glycol ◽  
Embryo Quality ◽  
Control Group ◽  
Follicular Growth ◽  
Short Term ◽  
Embryo Production ◽  
Blastocyst Quality ◽  
In Vitro Embryo Production ◽  
Ovarian Follicular Growth

Rapid genetic improvement in cattle requires the production of high numbers of embryos of excellent quality. Increasing circulating insulin and/or glucose concentrations improves ovarian follicular growth, which may improve the response to superovulation. The measurement of anti-Müllerian hormone (AMH) can help predict an animal’s response to superovulation treatment. The aim of the present study was to investigate whether increasing circulating insulin concentrations, through propylene glycol (PG) drenches, could improve in vitro embryo production in oestrus-synchronised superovulated heifers with different AMH profiles. Holstein heifers were grouped according to pre-experimental AMH concentrations as low (L) or high (H). The PG drench increased circulating insulin and glucose concentrations and reduced β-hydroxybutyrate and urea concentrations compared with the control group. AMH was a good predictor of follicle and oocyte numbers at ovum pick-up (OPU), and of oocyte and embryo quality (AMH H > AMH L). PG in the AMH H group increased the number of follicles and blastocyst quality above that in the control group, but did not improve these parameters in the AMH L group. These results indicate that short-term oral PG supplementation modifies an animal’s metabolic milieu and is effective in improving in vitro embryo production, after superovulation–OPU, more markedly in heifers with high rather than low AMH concentrations.

231 EFFECTS OF DIETARY PROPYLENE GLYCOL ON FOLLICULAR FLUID AND OVUM PICK UP IN VITRO EMBRYO PRODUCTION IN GROWTH-RESTRICTED HEIFERS WITH TWO PROFILES OF ANTI-MÜLLERIAN HORMONE

2015 ◽  
Vol 27 (1) ◽  
pp. 205
Author(s):  
G. Gamarra ◽  
C. Ponsart ◽  
S. Lacaze ◽  
B. Le Guienne ◽  
P. Humblot ◽  
...  
Keyword(s):  
Propylene Glycol ◽  
Follicular Fluid ◽  
Plasma Concentrations ◽  
Oestrous Cycle ◽  
Water Control ◽  
Embryo Production ◽  
Blood Samples ◽  
Metabolic Hormones ◽  
In Vitro Embryo Production

Fertility and embryo quality can be improved in cattle by using diets that induce a programmed modulation of circulating insulin concentrations. The aim of this study was to test whether the daily oral administration of propylene glycol (PG) could modify metabolite and hormone plasma and follicular fluid concentrations and improve in vitro embryo production in superovulated growth-restricted heifers (600 g day–1). Sixteen Holstein heifers were grouped according to their pre-experimental anti-Müllerian hormone (AMH) plasma concentrations: low (L = 1–80 pg mL–1; n = 7) or high (H: >150 pg mL–1; n = 9). Heifers received a single daily drench from Day 1 to Day 9 of an oestrous cycle [first cycle, 400 mL of water (control) and second cycle, 400 mL of PG]. Serial jugular blood samples were collected on Day 7 of each cycle to monitor plasma insulin, glucose, and β-hydroxybutyrate (BHB) concentrations in relation to the drench. Blood samples were also collected to measure insulin-like growth factor-1 (IGF1) and progesterone (P4) concentrations on Days 0, 2, 5, 7, and 9 of the oestrous cycle. Follicular fluid was collected on Day 9 to measure insulin and IGF1 concentrations. Ovarian ultrasonography was performed on Days 2 and 5 to count follicles between 2 and 8 mm in diameter and estimate their size. After ovum pickup (OPU) performed following superovulation on Day 5 of the oestrous cycle, oocytes were matured and fertilized in vitro, then embryos were cultured for 7 days. Propylene glycol increased plasma concentrations of insulin and glucose and reduced BHB in both groups of heifers compared with control. It also increased IGF1 concentrations on Days 5 and 7 in AMH L heifers and on Days 2, 5, and 7 in AMH H heifers, and reduced P4 concentrations on Days 5 and 9 of the oestrous cycle in all heifers. In follicular fluid, there was no difference in insulin concentrations between groups, but PG increased IGF1 concentrations in all heifers. In ovaries, PG increased the number of small follicles (2–3 mm) and total follicles on Day 2 of the cycle in all heifers, and medium follicles (4–8 mm) and total follicles on Day 5 in AMH H heifers. Propylene glycol improved the in vitro embryo development rate (total number of embryos/number of fertilized oocytes) in all heifers (AMH L: control, 37.9% v. PG, 50.0%; P < 0.05; AMH H: control, 36.4% v. PG, 48.3%; P < 0.05). In AMH H, the number of grade 1 blastocysts was increased by PG (control, 5.2 ± 1.0 v. PG, 8.9 ± 1.0; P < 0.01), whereas there was no difference between treatments in AMH L heifers (control, 1.9 ± 1.1 v. PG, 3.2 ± 1.1; P > 0.05). These results indicate that short-term oral PG supplementation affects the concentrations of metabolites and metabolic hormones in blood and IGF1 concentrations in follicular fluid. PG administration is effective in improving in vitro embryo production more markedly in heifers with high AMH compared with low AMH endocrine levels.

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

Animals ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 763 ◽  
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.

133 Test of minimum-intervention protocols for optimizing in vitro embryo production in bison

2019 ◽  
Vol 31 (1) ◽  
pp. 192
Author(s):  
M. L. Zwiefelhofer ◽  
E. M. Zwiefelhofer ◽  
S. X. Yang ◽  
S. Maeda ◽  
J. Singh ◽  
...  
Keyword(s):  
Control Group ◽  
Nominal Data ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Chi Squared ◽  
In Vitro Embryo Production ◽  
Categorical Distribution ◽  
Long Acting ◽  
Parks Canada

The study was done to determine whether minimal handling protocols for ovarian synchronization and ovarian superstimulation may be used to increase in vitro embryo production in bison. Ultrasound-guided cumulus-oocyte complex (COC) collection was done in a group of bison (n=23; random start) during the anovulatory season to synchronize new follicular wave emergence. The COC were classified morphologically (compact-good and -regular, expanded, denuded, degenerate) but not processed further. At the time of COC collection (Day 0), bison were assigned randomly to 3 groups and given 5mL of saline IM (non-superstimulated controls; n=11), 10 Armour units of pFSH (Antrin R10, Kyoritsu Seiyaku Corp., Tokyo, Japan) in 5mL of saline IM once per day from Day 0 to 2 (regular FSH; n=5), or 30 armour units of a sustained-release form of pFSH (Antrin R10Al, Kyoritsu Seiyaku Corp.) in 5mL of saline SC on Day 0 (long-acting FSH; n=7). On Day 4, a second COC collection was performed. Only compact COC were processed. The COC were matured in vitro for 25 to 28h at 38.8°C, fertilized (2×106 sperm mL−1) and co-incubated at 38.8°C in 5% CO2 for 18h. Presumptive zygotes were denuded and cultured at 38.8°C in 5% O2, 5% CO2 and 90% N2. Nominal data were compared by t-test and analysis of variance. Binomial data were compared among groups by chi-squared. There was no difference between the first (random) COC collection (n=23) and second collection (n=11 non-superstimulated controls) in the total number of follicles detected, but the distribution among size categories (3-4, 4-8, and &gt;8mm) differed, i.e. fewer in the 3 to 4mm category at the time of the second COC collection (12.2±1.0v. 8.1±1.4; P&lt;0.05). In the nonstimulated control group, there were no differences between the first and second COC collections in the number of follicles aspirated (12.7±1.0v. 10.4±1.5), number of COC collected (7.7±0.9v. 5.3±1.3), or in the categorical distribution of COC. At the second COC collection, the number of follicles in the &gt;8mm category was greater in the regular FSH group than in the control or long-acting FSH groups (2.8±0.5v. 1.1±0.3, and 1.9±0.4, respectively; P&lt;0.05), but no differences were detected in the number of follicles aspirated, COC collected, or in the categorical distribution of COC. The cleavage rate (of total oocytes submitted to in vitro maturation), recorded 2 days after IVF, was higher in the control group than in either the regular FSH or long-acting FSH groups [25/35 (71.4%), 7/28 (25.0%), 8/35 (22.8%); P&lt;0.0001]. The freezable embryo production rate, recorded 7 days after IVF, was greater in the control group than in the regular FSH or long-acting FSH groups [19/35 (54.3%), 5/28 (17.9%), 5/35 (14.3%); P&lt;0.01]. In conclusion, minimal-handling interventions used in the present study to increase embryo production in bison were not effective, likely as a result of the timing, frequency, and duration of superstimulation. A random start resulted in greater COC collection than collection 4 days after ovarian synchronization, and embryo production rates were greater in non-superstimulated bison. This work was supported by Parks Canada and Saskatchewan ADF. Antrin products donated by Kyoritsu Seiyaku Corp., Japan.

Pregnancy obtained in a late gestational mare by in vitro embryo production

2019 ◽  
Vol 31 (12) ◽  
pp. 1926
Author(s):  
Lino Fernando Campos-Chillon ◽  
Jan Martin ◽  
Joy L. Altermatt
Keyword(s):  
Transvaginal Ultrasound ◽  
Experimental Studies ◽  
Late Gestation ◽  
Follicular Growth ◽  
Ultrasound Guided ◽  
Embryo Production ◽  
In Vitro Embryo Production

Recently, the demand for invitro embryo production in the horse has increased worldwide. Most clinical transvaginal ultrasound-guided ovum pick-up (OPU) procedures are performed in non-pregnant donor mares, and few experimental studies have described invitro embryo production from oocytes of pregnant donors 21–150 days in gestation. This report discusses OPU, follicular growth and invitro embryo production in a pregnant mare during late gestation.

scholarly journals Effect of deslorelin acetate treatment in oocyte recovery and in vitro embryo production in domestic cats

2016 ◽  
Vol 19 (10) ◽  
pp. 1091-1095
Author(s):  
Camila Louise Ackermann ◽  
Eduardo Trevisol ◽  
Leticia Ferrari Crocomo ◽  
Tatiana da Silva Rascado ◽  
Rodrigo Volpato ◽  
...  
Keyword(s):  
Treated Group ◽  
Control Group ◽  
Domestic Cats ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Oocyte Recovery ◽  
Significant Difference ◽  
In Vitro Embryo Production ◽  
Blastocyst Rate

Objectives The present study investigated the effect of contraceptive treatment with deslorelin acetate on in vitro embryo production and oocyte recovery in domestic queens. Methods Twenty-one mature domestic cats were used. Eleven queens (treated group) and one tom were kept in an experimental cattery, and 10 queens were privately owned (control group). When in interestrus or diestrus (day 0) a deslorelin acetate implant (Suprelorin, 4.7 mg/animal) was inserted into the subcutaneous tissue of the interscapular region in all queens in the treated group. After 6 months of treatment, all animals were ovariohysterectomized, and the ovaries were used for in vitro embryo production. Percentage of cleavage was determined 18 h after oocyte insemination and blastocyst formation was assessed on the eighth day of culture. The rate of cumulus-oocyte complexes (COCs) recovery was analyzed by an unpaired t-test. The cleavage and blastocyst rates were expressed as percentages and analyzed by Fisher’s exact test. All analyses were performed using GraphPad Prism v5.0, with P <0.05 set as the level of significance. Results In the treated group, we recovered 8.3 ± 1.15 grade I COCs per queen; the cleavage rate was 60% and the blastocyst rate was 36%. In the control group, we recovered 18.4 ± 3.21 grade I COCs per queen; the cleavage rate was 55.97% and the blastocyst rate was 34%. Forty percent of treated females did not produce any blastocysts. In the treated group, we observed a significant decrease in COC recovery. Although there was no significant difference in cleavage and blastocyst rates between groups, 40% of treated females did not produce any blastocysts. Conclusions Recovery of grade I COCs is negatively affected by deslorelin treatment in domestic cats. Regarding embryo production, new studies are still necessary to evaluate the success of this technique owing to the individual effect caused by deslorelin acetate.

234 HORMONAL FOLLICLE STIMULATION IN HOLSTEIN COWS FOR IN VITRO EMBRYO PRODUCTION USING SPERM SORTED BY FLOW CYTOMETRY

2016 ◽  
Vol 28 (2) ◽  
pp. 248 ◽  
Author(s):  
L. Ferré ◽  
Y. Bogliotti ◽  
J. Chitwood ◽  
M. Kjelland ◽  
P. Ross
Keyword(s):  
Transvaginal Ultrasound ◽  
Prostaglandin F2α ◽  
Holstein Cows ◽  
Control Group ◽  
Hatching Rate ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
Group 2 ◽  
Group 1

Transvaginal ultrasound needle-guided ovum pick-up (OPU) and in vitro embryo production (IVP) offer a reliable alternative to conventional embryo transfer to produce offspring. The success of OPU/IVP greatly depends on the number and quality of retrieved oocytes. The aim of this study was to compare OPU/IVP performance from stimulated Holstein cows. Holstein (Bos taurus) >8-year-old pluriparous open dry cows (n = 28) were used for OPU as oocyte donors. Follicular waves in all groups were synchronized by gonadotropin-releasing hormone (GnRH), prostaglandin F2α (PGF), and CIDR administrated on Day 0, followed by stimulation treatments 48 h later. No pre-synch was used. Total hormone dosage were administrated as follows: Group 1: pFSH = 180 mg (Folltropin, Bioniche, Belleville, ON, Canada; n = 7), Group 2: pFSH/LH = 500 IU (Pluset, Calier, Barcelona, Spain; n = 7), Group 3: eCG = 1500 IU (eCG, Biogénesis-Bagó, Buenos Aires, Argentina; n = 7) and Group 4: Control (n = 7), no stimulation. All injections were performed intramuscularly (i.m.) twice a day, during three days. OPU was performed 48 (Group 1) or 24 h (Group 2 and 3) after the last injection. The control group received saline solution i.m. Follicles were classified according to diameter in 4 categories: small (2–5 mm); medium (6–9 mm); large (10–14 mm) and extra large (>15 mm). A Mindray DP-30 Vet (Mindray Medical, Shenzhen, China) was equipped with a micro-convex transducer 5.0- to 8.5-MHz probe along with a disposable 21G needle. The OPU flow rate was 15 mL min–1. Retrieved oocytes were classified according to IETS guidelines as viable (grade 1 + 2) and non-viable (grade 3 + 4). The IVP protocol was according to that in Reprod. Fertil. Devel. (2004, 16, 253). Fertilization (Day 0) was carried out using female sex-sorted semen selected with a discontinuous density gradient (PureSperm, Nidacon, Mölndal, Sweden) and diluted to 1 × 106 sperm mL–1. ANOVA was used for comparisons of mean values and a chi-squared test was used for proportions. Results are presented in the Table 1. In conclusion, pFSH stimulation before ovum pick-up in Holstein cows increased the number of collected and viable oocytes, cleavage, embryo development, and hatching rates in comparison to other follicle stimulation hormones and non-stimulation. A cost-benefit analysis of these methods could be valuable in order to inform whether or not a stimulation protocol is necessary for a commercial IVP operation. Table 1.Numbers of follicles, collected and viable oocytes, cleavage rate, blastocysts and hatching rate

30 Vitrification of Equine Embryos: Application in a Commercial Cloning Program

2018 ◽  
Vol 30 (1) ◽  
pp. 154
Author(s):  
G. Vichera ◽  
R. Jordan ◽  
V. Arnold ◽  
D. Dobler ◽  
R. Olivera
Keyword(s):  
Breeding Season ◽  
Production Efficiency ◽  
Embryo Quality ◽  
Donor Cell ◽  
Blastocyst Stage ◽  
Pregnancy Rates ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
The One

During a commercial horse cloning program, a critical point is the availability and maintenance of suitable recipient mares for a large quantity of embryo transfers. Usually, pregnancy rates and viable births off the breeding season decrease significantly, whereas the rate of in vitro embryo production remains constant. For this reason, an efficient vitrification system allows continuous embryo production throughout the year with the advantage of doing the transfers only during the breeding season. The vitrification technique evaluated in this study was the one described by Kuwayama et al. (2007 Theriogenology 67, 73-80). By using this method, we compared post-warming recovery efficiency, pregnancy rates, and viable foaling rates in 2 experimental groups: cloned blastocysts vitrified off-season and transferred in breeding season (VC, n = 337), and non-vitrified cloned blastocysts also transferred in breading season (no-VC, n = 516). To achieve this, equine oocytes were collected from slaughterhouse ovaries, matured, enucleated, and fused to a donor cell according to Olivera et al. (2016 PLoS One 11, e0164049, 10.1371/journal.pone.0164049). The reconstructed embryos (RE) were cultured in a well-of-the-well system by adding 3 RE per well for 7 to 8 days to reach the blastocyst stage, at which they were vitrified as mentioned above. During the breeding season, blastocysts were warmed and transferred in couples in a single cycling receptive mare. Pregnancies were confirmed by transrectal ultrasonography 15 days post-transfer. All variables were analysed by Fisher test (P < 0.05). The warming recovery rate was 91% (308/337) for cloned blastocysts. In addition, pregnancy and viable birth rates were similar for the VC and no-VC groups: 15.6% (24/154) v. 16.7% (43/258) for pregnancy rates, respectively, and 37.5% (9/24) v. 37.2% (16/43) for foaling rates, respectively. In summary, 9 viable cloned foals were obtained with off-season embryos warmed and transferred during the breeding season, showing that vitrification did not affect embryo quality. Hence, the proposed strategy provides the ability to maximize production efficiency of equine clones by generating a large number of pregnancies without stopping in vitro embryo production at any time of the year.

scholarly journals Evidence of haptoglobin in the porcine female genital tract during oestrous cycle and its effect on in vitro embryo production

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Francisco A. García-Vázquez ◽  
Carla Moros-Nicolás ◽  
Rebeca López-Úbeda ◽  
Ernesto Rodríguez-Tobón ◽  
Ascensión Guillén-Martínez ◽  
...  
Keyword(s):  
Western Blot ◽  
Female Genital Tract ◽  
Oestrous Cycle ◽  
Control Group ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Or Efficiency ◽  
In Vitro Embryo Production ◽  
Proteomic Analyses

AbstractRecent evidence supports involvement of the acute phase protein haptoglobin in numerous events during mammalian reproduction. The present study represents an in-depth investigation of haptoglobin expression and secretion in the porcine oviduct and uterus, and assesses its effect on porcine in vitro embryo production. A systematic study was made of sows in different oestrous stages: late follicular, early luteal and late luteal stages. Relative haptoglobin mRNA abundance was quantified by RT-qPCR. In addition, expression of the protein was analysed by immunohistochemistry and the results were complemented by Western-blot and proteomic analyses of the oviductal and uterine fluids. In vitro porcine fertilization and embryo culture were carried out in the presence of haptoglobin. The results indicate that haptoglobin mRNA expression in the porcine oviduct and uterus is most abundant during the late luteal stage of the oestrous cycle. By means of Western blot and proteomic analyses haptoglobin presence was demonstrated in the oviduct epithelium and in the oviductal and uterine fluids in different stages of the oestrous cycle. The addition of haptoglobin during gamete co-incubation had no effect on sperm penetration, monospermy or efficiency rates; however, compared with the control group, blastocyst development was significantly improved when haptoglobin was present (haptoglobin: 64.50% vs. control: 37.83%; p < 0.05). In conclusion, the presence of haptoglobin in the oviduct and uterus of sows at different stages of the oestrous cycle suggests that it plays an important role in the reproduction process. The addition of haptoglobin during in vitro embryo production improved the blastocyst rates.

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