125 Influence of length of porcine follicle-stimulating hormone (p-FSH) treatment before ovum pickup on ovarian response and invitro embryo production in Holstein heifers

2021 ◽  
Vol 33 (2) ◽  
pp. 170
Author(s):  
J. C. L. Motta ◽  
R. V. Sala ◽  
V. A. Absalón-Medina ◽  
V. C. Fricke ◽  
M. Dominguez ◽  
...  
Keyword(s):  
Treatment Period ◽  
Recovery Rate ◽  
Body Condition Score ◽  
Ovarian Follicle ◽  
Ovarian Response ◽  
Medium Size ◽  
P Value ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Blastocyst Rate

Ovarian follicle stimulation with exogenous FSH before ovum pickup (OPU) in Bos taurus females is a common practice to increase invitro embryo production (IVP). The optimal stimulatory period length for OPU-IVP, however, has not been definitively ascertained. The objective of the present study, therefore, was to determine the effect of length of the superstimulatory treatment period before OPU on ovarian response and IVP in Holstein heifers. Nonpregnant heifers (n=57) 13.8±0.2 months of age with moderate body condition score (3.0±0.1; scale 1 to 5) were assigned in a completely randomised design to one of the following experimental groups: FSH2d: 200mg of p-FSH (Folltropin-V®, Vetoquinol) distributed in four injections (60, 60, 40, and 40mg) of FSH 12h apart; FSH3d: 200mg of p-FSH distributed in six injections (40, 40, 40, 40, 20, and 20mg) of FSH 12h apart. Timing of follicular wave emergence was synchronized by dominant follicle removal 36h before the first p-FSH injection in all heifers. An intravaginal progesterone (P4) implant (1.38g of P4 CIDR®, Zoetis) was inserted at the time of the first p-FSH injection and removed at the time of OPU, which occurred in all heifers at 44h (Nivet et al. 2012 Reproduction 143, 165-171; https://doi.org/10.1530/REP-11-0391) after the last p-FSH injection. Additionally, follicle number was determined at OPU and classified as small (<6mm), medium (6–10mm) or large (>10mm). Oocytes from follicles of different sizes were pooled by heifer at OPU and then classified and subjected to IVP procedures. Differences between treatment groups were evaluated using generalized linear mixed models (SAS 9.4; SAS Institute Inc.) and data are presented as mean±s.e.m. (Table 1). Lengthening the FSH treatment period resulted in a greater (P=0.01) number of large follicles; however, the number of small, medium-size, and total follicles was not different between groups. Number of total recovered oocytes, viable oocytes, cleaved oocytes, as well as recovery rate, percent viable oocytes, and cleavage rate were not different (P>0.2) between groups. Similarly, the number of blastocysts produced per heifer and blastocyst rate were not different (P>0.9) among groups. In conclusion, lengthening the period of FSH treatment by 1 day increased the number of large follicles at OPU but did not improve overall ovarian response, oocyte recovery, or embryo production. Table 1. Ovarian response and IVP of heifers treated with p-FSH during 2 or 3 days before ovum pickup Response FSH2d (n=28) FSH3d (n=29) P-value Small follicles (n) 5.9±0.6 5.7±0.8 0.83 Medium follicles (n) 17.0±2.4 12.9±1.6 0.18 Large follicles (n) 2.5±0.5 4.5±0.6 0.01 Total follicles (n) 25.4±2.6 23.1±1.8 0.60 Total oocytes (n) 17.0±2.5 13.0±1.2 0.23 Recovery rate (%) 62.6±3.7 56.9±3.1 0.26 Viable oocytes (n) 14.4±2.0 11.3±1.1 0.30 Viable oocytes (%) 85.0±2.4 88.0±3.0 0.31 Cleaved oocytes (n) 8.7±1.6 7.1±1.2 0.62 Cleavage rate (%) 54.7±5.7 54.1±5.7 0.96 Blastocysts (n) 3.2±0.6 3.2±0.7 0.98 Blastocyst rate (%) 20.6±4.0 22.4±3.7 0.97

71 Effect of follicle-stimulating hormone dose and circulating progesterone before ovum pickup and invitro embryo production in pregnant Holstein heifers

2021 ◽  
Vol 33 (2) ◽  
pp. 143
Author(s):  
C. Hayden ◽  
R. V. Sala ◽  
D. C. Pereira ◽  
L. C. Carrenho-Sala ◽  
M. Fosado ◽  
...  
Keyword(s):  
Body Condition Score ◽  
Medium Size ◽  
P Value ◽  
Limited Information ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Treatment Groups ◽  
Standard Procedures ◽  
Condition Score ◽  
Blastocyst Rate

Superstimulation with exogenous FSH before ovum pickup for invitro embryo production (IVP) improves embryo production. However, there is limited information on different FSH dose regimens and the effect of progesterone (P4) concentrations on IVP. Thus, the objective of the present study was to evaluate the effect of FSH dose and circulating P4 on ovarian superstimulation responses, before ovum pickup (OPU) on IVP in pregnant heifers. Pregnant Holstein heifers (n=47) 19.5±0.3 months of age with moderate body condition score (3.3±0.03; scale 1 to 5) were randomly assigned to 2 treatment groups, p-FSH (Folltropin-V, Vetoquinol) 160mg (FSH160) or p-FSH 300mg (FSH300), in a crossover design. Two sessions of OPU were performed on each animal at 48 and 62 days of gestation, with a washout interval between sessions of 14 days. Dominant follicle removal (DFR) was utilised to synchronize follicular wave emergence, and FSH treatments were initiated 36h later. Heifers in both groups received a total of 4 FSH injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3, and 32.0mg) or FSH300 (90.0, 80.0, 70.0, and 60.0mg). All heifers underwent OPU 40h after the last FSH treatment. Blood samples were collected for P4 quantitation at OPU and number of small (<6mm), medium (6–10mm), and large (>10mm) follicles were determined before OPU. Cumulus–oocyte complexes (COCs) recovered were processed and invitro embryo production was performed using standard procedures. Differences between treatment groups were evaluated using generalized linear mixed models, and data are presented as mean±s.e.m. (Table 1). Heifers treated with 300mg of p-FSH had a greater (P<0.05) number of medium-size follicles, total number of follicles, total COCs recovered, number of viable COCs, and number of blastocysts produced. In addition, heifers treated with 300mg of p-FSH had a greater cleavage rate (P=0.03). Circulating P4 was on average 10.1±0.4ng mL−1 and did not differ between gestation days (P=0.12). Furthermore, circulating P4 did not affect (P>0.10) any of the outcomes evaluated, nor was there an FSH dose by circulating P4 interaction (P>0.05). In conclusion, increasing the dose of p-FSH before OPU from 160 to 300mg resulted in an increase in the total number of follicles, number of oocytes, and number of blastocysts produced in pregnant donors without changes in blastocyst rate, regardless of progesterone concentrations at the time of OPU. Table 1. Ovum pickup and invitro embryo production in pregnant heifers treated with different doses of p-FSH Item FSH160 FSH300 P-value Small follicles (n) 5.0±0.6 3.5±0.4 0.02 Medium follicles (n) 14.4±0.8 18.3±0.9 0.0003 Large follicles (n) 2.9±0.3 2.5±0.3 0.28 Total follicles (n) 22.3±0.7 24.3±0.9 0.04 Total oocytes (n) 12.8±0.7 14.7±1.0 0.02 Recovery rate (%) 56.2±2.1 59.7±2.7 0.20 Viable oocytes (n) 11.4±0.7 13.3±1.0 0.02 Viable oocytes (%) 89.9±1.7 88.0±2.4 0.22 Cleavage rate (%) 63.8±4.3 71.5±3.7 0.03 Cleaved oocytes (n) 7.2±0.7 9.3±0.9 0.14 Blastocysts (n) 2.7±0.4 3.6±0.7 0.02 Blastocyst rate (%) 22.0±3.3 22.4±3.6 0.74

147 Simplification of the follicle-stimulating hormone protocol for superovulation of the first follicular wave in sheep

2021 ◽  
Vol 33 (2) ◽  
pp. 181
Author(s):  
C. Brochado ◽  
B. J. Duran ◽  
J. C. L. Motta ◽  
J. D. Kieffer ◽  
A. Pinczak ◽  
...  
Keyword(s):  
Dose Group ◽  
Body Condition Score ◽  
Ovarian Response ◽  
Chorionic Gonadotrophin ◽  
P Value ◽  
Corpora Lutea ◽  
Embryo Production ◽  
Follicular Wave ◽  
Condition Score ◽  
Dose Number

The objective of the present study was to evaluate the effect of length of the FSH superstimulatory treatment on ovarian response and embryo production in sheep. Poll Dorset ewes (n=63) 3.2±0.2 years old weighing 58.5±1.5kg and with a body condition score of 2.7±0.1 (0=emaciated; 5=obese) were used during the transition from the breeding to the non-breeding season. All ewes received an ovarian superstimulatory treatment during the first follicular wave (Day 0 protocol). On Day −9 all ewes received a 0.3-g progesterone intravaginal device (CIDR, Zoetis) for 6 days. On Day −3, the CIDR was removed and all ewes were administered 125μg of cloprostenol sodium intramuscularly (IM) (PGF, estroPLAN, Parnell) and 200IU of equine chorionic gonadotrophin (eCG)+100IU of human chorionic gonadotrophin (hCG) IM (P.G. 600, Merck). Thirty-six hours after CIDR removal (Day −1.5) ewes were administered 100μg of gonadorelin acetate IM (gonadotrophin-releasing hormone, GnRH; Gonabreed, Parnell). Superstimulatory treatments were initiated on Day 0 (84 h after CIDR removal) with a total of 240mg of NIH-FSH-P1 (12 mL, Folltropin, Vetoquinol), and a new CIDR was used from the first to the last FSH dose (6-dose=3 days; 8-dose=4 days). Ewes were randomised to receive the total dose of FSH distributed in eight decreasing doses (8-dose group, n=33) or six decreasing doses (6-dose group, n=30) twice daily 12h apart. Two doses of PGF were administered with the last two doses of FSH, and GnRH was administered 12h after the last FSH administration. Intrauterine AI with fresh semen was performed by laparoscopy 16h after GnRH administration. Ovarian response was determined by laparoscopy 6 days after AI and ova/embryos were collected surgically in ewes with >2 corpora lutea (CL). Data were analysed using generalized linear mixed models (SAS 9.4; SAS Institute Inc.) and presented as mean±s.e.m. in Table 1. The percentage of responding donors (>2 CL) was 93.3% (28/30) and 78.8% (26/33; P=0.5) for the 6-dose and 8-dose groups, respectively. The number of CL tended (P=0.06) to be lesser in ewes in the 6-dose than the 8-dose group. However, no differences (P>0.10) were found in total ova/embryo, fertilized ova, transferable embryos, or freezable embryos. In conclusion, despite a tendency for a greater ovarian response in ewes treated with 8 doses of FSH, embryo yield did not appear to differ after either 6 or 8 doses of FSH administered during the first follicular wave in sheep. Table 1. Ovulatory response and embryo production in responding ewes (>2 corpora lutea) superstimulated with either 6 or 8 doses of FSH during the first follicular wave Outcome Treatment P-value 6-dose 8-dose Number of ewes 28 26 Corpora lutea (n) 14.2±1.7 20.3±2.8 0.06 Total ova/embryos (n) 9.0±1.3 10.9±1.8 0.38 Fertilized ova (n) 8.1±1.4 10.2±1.9 0.34 Fertilized ova (% per ewe) 89.0±5.0 81.7±7.0 0.40 Degenerate embryos 1.3±0.4 2.0±0.8 0.89 Transferable embryos (Grade 1–3) 6.8±1.2 8.2±1.8 0.52 Transferable embryos (% per ewe) 85.7±4.4 75.4±8.4 0.46 Freezable embryos (Grade 1–2) 6.6±1.1 8.2±1.8 0.43 Freezable embryos (% per ewe) 80.5±5.6 75.4±8.4 0.61

176 Synchronisation of follicle wave emergence prior to superstimulation with purified FSH for ovum pickup affects blastocyst rate in pregnant Holstein heifers

2020 ◽  
Vol 32 (2) ◽  
pp. 215
Author(s):  
L. Carrenho-Sala ◽  
M. Fosado ◽  
R. Sala ◽  
E. Peralta ◽  
D. Pereira ◽  
...  
Keyword(s):  
Mixed Models ◽  
Ovarian Response ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Treatment Groups ◽  
Follicular Wave ◽  
The Mean ◽  
Blastocyst Rate ◽  
Oocyte Donors ◽  
Timing Of Initiation

The timing of initiation of superstimulatory treatments relative to follicle wave emergence has been shown to affect ovulatory response and invivo embryo production. The significant increase of invitro embryo production (IVP) and the possibility of using pregnant animals as oocyte donors has created the need to optimise superstimulatory treatments for IVP in pregnant cattle. Thus, the objective of the present study was to evaluate the effect of synchronisation of follicle wave emergence before superstimulation for ovum pickup (OPU) and IVP in pregnant heifers. Pregnant (47-62 days of gestation) Holstein heifers (n=28) 19.5±0.3 months of age were assigned in a completely randomised design to one of two groups: synchronisation of follicular wave emergence by dominant follicle removal (DFR; all follicles >6mm) or untreated control (no DFR). Superstimulatory treatments were initiated 36h after DFR or at random stages of the follicular wave in the no-DFR group and consisted of the administration of 160mg of purified FSH (Folltropin-V, Vetoquinol) over four injections 12h apart as follows: 48.0, 42.7, 37.3, and 32.0mg. Ovum pickup was performed in all heifers 40h after the last purified FSH injection. Heifers were subjected to OPU for oocyte recovery, and the number of follicles was determined. Recovered oocytes were processed in groups by treatment, and IVP was performed. Differences between treatment groups were evaluated using generalised linear mixed models. Results are presented in Table 1 and are expressed as means±s.e.m. for data collected at the time of OPU or as proportions for embryo production results. The number of small follicles (<6mm) at the time of OPU was greater in the no-DFR group than in the DFR group (P=0.04). Conversely, there were no differences between treatments in the number of medium follicles (6-10 mm; P=0.17), large follicles (>10 mm; P=0.11), total follicles (P=0.93), total number of recovered oocytes (P=0.4), or number of viable oocytes (P=0.53). The mean oocyte percentage recovery rate was not different between heifers in the DFR (53.6±4.7%) and no-DFR (56.5±4.7%) groups (P=0.52). Both cleavage and blastocyst rate were greater (P<0.008) in the DFR group than in the no-DFR group; as a result, the number of transferable embryos per animal was 5.6 in the DFR group and 2.8 in the no-DFR group. In summary, initiation of superstimulatory treatments at the time of follicle wave emergence improves cleavage and blastocyst rates, thus leading to greater embryo production. Table 1.Ovarian response and embryo production in pregnant heifers superstimulated with or without synchronisation of follicle wave emergence Variable DFR No DFR Small follicles, n 8.1±1.2A 12.1±1.8B Medium follicles, n 18.3±1.3 13.7±2.0 Large follicles, n 2.4±0.6 1.4±0.4 Total follicles, n 28.8±1.4 27.2±2.2 Total oocytes, n 15.4±1.5 16.0±1.9 Viable oocytes, n 13.7±1.5 13.4±1.8 Cleavage rate,% (n) 77.1 (192)A 64.4 (188)B Blastocyst rate,% (n) 40.6 (192)A 20.7 (188)B A,BMeans within a row with different superscripts differ (P<0.05).

149 EFFECTS OF FSH ADDED IN DEFINED MEDIUM MATURATION ON THE PRE-IMPLANTATION DEVELOPMENT AND EXPRESSION OF Bax, Bcl-2, AND CONNEXIN 43 GENES IN BOVINE IVP BLASTOCYSTS

2010 ◽  
Vol 22 (1) ◽  
pp. 233
Author(s):  
L. V. M. Gulart ◽  
L. Gabriel ◽  
L. P. Salles ◽  
G. R. Gamas ◽  
D. K. Souza ◽  
...  
Keyword(s):  
Internal Control ◽  
Connexin 43 ◽  
Calf Serum ◽  
Defined Medium ◽  
Culture Conditions ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Blastocyst Rate

FSH at low concentrations affect embryo production. In vitro culture conditions also affect embryo production and embryonic expression of genes and alter oocyte competence to produce embryos. The search for better and less variable culture conditions simulating those in vivo has led to the development of several systems of oocyte in vitro maturation culture. To compare the efficiency of the systems of MIV we utilized 4 groups: (1) TCM-199 control; (2) α-minimal essential medium (MEM); 3) α-MEM + 1 ng of FSH; 4) α-MEM+ 10 ng of FSH. The medium of Group 1 is non-defined by the presence of fetal calf serum (10%). Groups 2, 3, and 4 are defined and polyvinyl alcohol (1%) was used as a macromolecule. Porcine FSH (1 IU mg-1) was used at 1 and 10 ng mL-1 and at 100 ng in defined and non-defined medium, respectively. Bovine ovaries were collected at an abbatoir. Oocytes (n = 1718) with homogeneous cytoplasm and with more than 3 layers of granulosa cells were used. Mature oocytes from the 4 treatments (11 replicates of each treatment) were inseminated with frozen-thawed, motile sperm separated by Percoll, using Sperm TALP HEPES medium. Presumptive zygotes with up to 2 or 3 layers of cumulus cells were cultured in 50-mL drops of SOF medium, supplemented with 10% FCS and 1 mg mL-1 BSA under mineral oil in a humid 5% CO2 atmosphere at 38.5°C after. Cleavage rate was evaluated 72 h post-insemination (hpi), and blastocyst rate was evaluated 168-192 hpi. Cleavage and blastocyst rates were calculated on the basis of number of presumptive zygotes. The expression of the following genes (Bax, Bcl-2, and conexin 43) was evaluated in blastocysts by RT-PCR. One-way ANOVA was used to compare blastocyst number. There was no difference in the proportion of embryos with more than 8 blastomeres in all groups tested, indicating that the rate of development during the first 72 hpi was similar for oocytes matured in chemically defined medium and for oocytes matured in medium containing serum. Bax is a pro-apoptotic marker and Bcl-2 an antiapoptotic marker. Connexin 43 (Cx43) may be a marker of embryo competence. Glyceraldehyde 3-phosphate dehydrogenase was used as internal control. The Bax gene was not expressed in any group. The Bcl-2 and Cx43 genes were expressed, mainly in the α-MEM 10. Although no differences were observed in blastocyst rate among the groups (30% to 40%), the strong expression of Bcl-2 and of Cx43 on the group containing 10 ng mL-1 of FSH may indicate that FSH could improve embryo quality under defined conditions. The authors thank FAP-DF, CNPq, FUNPE, FINATEC, CAPES, and Biovitro Tecnologia de Embrioes Ltda, for laboratory assistance and grants, and Frigorifico Ponte Alta, Brasília-DF, for supplying bovine ovaries.

145 Comparison of single to multiple injections of follicle-stimulating hormone before ovum pickup in Holstein heifers: Oocyte recovery and embryo production

2021 ◽  
Vol 33 (2) ◽  
pp. 180
Author(s):  
D. G. B. Demetrio ◽  
J. F. Hasler ◽  
M. Oliveira ◽  
C. G. B. Demetrio ◽  
J. C. Fonseca ◽  
...  
Keyword(s):  
Time Synchronization ◽  
Random Effect ◽  
Oocyte Quality ◽  
Follicular Growth ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Multiple Injections ◽  
Oocyte Recovery ◽  
Blastocyst Rate ◽  
Stimulation Of

The demand for invitro-produced embryos from heifers with high genetic merit has increased over time. Synchronization and stimulation of follicular growth before ovum pickup (OPU) has been used to improve oocyte quality and, consequently, embryo production. Multiple injections involve extra labour and stress for both personnel and cattle. The release of FSH can be prolonged by using 0.5% hyaluronan (HA) as a diluent, allowing a decrease in the number of injections. The objective of this study was to compare oocyte recovery and embryo production between single or multiple injections of FSH before OPU of Holstein heifers. During April and May 2020, 20 Holstein heifers (8 to 15 mo old) from Ruann Dairy (Riverdale, CA) were randomly divided and submitted to two different treatments (crossover design). Gonadotrophin-releasing hormone (GnRH; Fertagyl®, Merck, 129µg, IM) was given to synchronize the follicular wave emergence. Treatment 1×FSH consisted of a single intramuscular (IM) injection of 100mg of FSH (Folltropin®, Vetoquinol) 36h after GnRH. The FSH consisted of a 2.5-mL injection of 400mg of FSH diluted in 10mL of 0.5% HA. OPU was performed 48 to 50h after FSH. Treatment 5×FSH consisted of 100mg of FSH divided into 5 equal IM injections (10-14h intervals) 36h after GnRH. The FSH consisted of 5×1-mL injections of 400mg of FSH in 20mL of saline. OPU was performed 18 to 20h after the last FSH injection. All donors received both treatments at a 14-day interval and the recovered oocytes were fertilized with the same sexed female-sorted semen in both rounds. OPU, oocyte classification, IVM, IVF, and culture (IVC) were performed as described by Demetrio et al. (2020 Anim. Reprod. 17, e20200053). All oocytes went into IVM, except for degenerated oocytes. The number of 4-cell (or more) embryos on Day 3 of IVC divided by the number of oocytes in IVC after IVF is defined as the cleavage rate. The number of blastocysts (early to hatched) on Day 7 of IVP divided by the number of oocytes in IVC after IVF is defined as the blastocyst rate. Poisson-normal (count data) and Logistic-normal (proportion data) models were used to analyse the data. Treatment, donor (random effect), and sire were included in the models. The results are summarized in Table 1. There were no differences between the two treatments on the number of oocytes recovered per OPU (total and grade 1 and 2), percentage of grade 1 and 2 oocytes, cleavage rate, blastocyst rate and number of embryos (total and grade 1). Oocyte recovery and embryo production are highly donor dependent. Stimulation of the follicular growth before OPU with one single injection of FSH diluted in 0.5% HA 36h after GnRH can be efficiently used for IVP in Holstein heifers, without decreasing the number of oocytes recovered and/or embryos produced with the advantage of reducing labour and stress of handling cattle. Table 1. Number and quality of oocytes and cleavage and blastocyst rates Treatment OPU Oocytes per donor Grade 1 and 2 oocytes (%) Cleavage rate (%) Blastocyst rate (%) Total embryos per OPU Grade 1 embryos per OPU 1×FSH 20 17.0 45.7 84 39.8 6.2 3.8 5×FSH 20 19.9 46.5 82 35.6 6.3 4.0

193 Effect of clinical metritis on oocyte recovery, oocyte quality, and early invitro developmental competence of embryos in Bos indicus dairy cattle

2020 ◽  
Vol 32 (2) ◽  
pp. 225
Author(s):  
M. Saleem ◽  
Z. Sarwar ◽  
M. Saad ◽  
I. Zahoor ◽  
N. Ahmad ◽  
...  
Keyword(s):  
Dairy Cattle ◽  
Recovery Rate ◽  
Body Condition Score ◽  
Bos Indicus ◽  
Oocyte Quality ◽  
Developmental Competence ◽  
Cleavage Rate ◽  
Healthy Group ◽  
Oocyte Recovery ◽  
Frozen Semen

Unhygienic practices at the time of parturition or AI lead to uterine infections. The uterine infections ultimately result in genetic drain by culling the elite animals. The invivo developmental competence of embryos is compromised in clinically metritic animals. The genetic potential of problematic females could be harvested by invitro embryo production (IVEP). Therefore, the objective of the present study was to evaluate the effect of clinical metritis on oocyte recovery, oocyte quality, and early invitro developmental competence of embryos in Bos indicus dairy cattle. This experiment was carried out from December 2017 to April 2018. Ovaries were collected from a local abattoir (Bos indicus; 5- to 8-year-old dairy cattle, body condition score 2.75±0.25, mixed parity). These ovaries (n=982) were divided into two groups: (1) clinically metritic (n=184), and (2) healthy (n=798), based upon the presence or absence of pus in the uterine lumen. Oocytes were aspirated from follicles using an 18G needle attached to a 10-mL syringe. Cumulus-oocyte complexes (COCs) were categorized into A, B, C, and D grades based on the number of layers of cumulus cells and integrity of ooplasm. The oocytes of grades A and B were subsequently transferred in groups (10/group) in four-well plates containing 100-μL droplets. The droplets with oocytes were covered with prewarmed mineral oil and incubated for 24h at 38.5°C, 5% CO2, and 95% relative humidity. The oocytes were evaluated for IVM on the basis of cumulus expansion. Frozen semen was thawed and prepared using the sperm swim-up procedure for each group. Spermatozoa and oocytes were incubated together for a period of 18h. The presumptive zygotes were invitro cultured for 4 days in a CO2 incubator under similar culture conditions. The cleavage rate, 4-cell, and 8-cell stages were recorded on Days 2, 3, and 4 after the day of insemination, respectively. Data on oocyte recovery, oocyte quality, IVM, cleavage rate, and 4-cell and 8-cell stages were analysed by Chi-squared test using SPSS software (version 20; IBM Corp.) for Windows. Results demonstrated that recovery rate was lower (63.8% vs. 71.7%; P<0.05) in clinically metritic compared with healthy cattle. Similarly, oocytes of grade A and B quality were lower (41.0% vs. 51.1%; P<0.05), whereas those of C and D quality were higher (59.0% vs. 48.9%; P<0.05) in clinically metritic compared with the healthy group. Moreover, 4-cell (38.2% vs. 54.8%) and 8-cell stage embryos (11.3% vs. 29.1%), were lower (P<0.05) in the clinically metritic compared with the healthy group, respectively. However, maturation rate and cleavage rate did not differ (P>0.05) between groups. In conclusion, metritis in slaughterhouse ovaries negatively affects oocyte recovery rate, oocyte quality, and early invitro developmental competence of embryos in Bos indicus dairy cattle.

PSX-A-23 Late-Breaking: Supplementation of Nerve Growth Factor (β-NGF) in the maturation medium and efficiency of in vitro embryo production in cattle

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 365-365
Author(s):  
Lucas Gonçalves ◽  
Muller C Martins ◽  
Natalia Arle ◽  
Rafaela T Torres ◽  
Luisa Migilo ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Oocyte Maturation ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Nerve Growth ◽  
Maturation Medium ◽  
In Vitro Embryo Production ◽  
Blastocyst Rate

Abstract The aim of this study was to evaluate the supplementation of Nerve Growth Factor (β-NGF) in the maturation medium in in vitro embryo production routines. Antral follicles were aspirated from ovaries of cows obtained from slaughterhouses and then oocytes were selected for quality (grades I and II) for in vitro maturation and subjected to 4 successive in vitro embryo production routines (IVEP). Supplementation of 100 ng of β-NGF was performed in the oocyte maturation medium 22 hours before in vitro fertilization. 48 hours after fertilization of the oocytes, an analysis was made of their cleavage rate by counting blastomeres with the aid of a stereoscopic microscope (cleavage rate = number of embryos / number of initial oocytes). Seven days after fertilization, the blastocyst rate was determined through the relation to the number of oocytes that started cleavage and reached this stage of development (blastocyst rate = number of blastocyst / number of oocytes that started cleavage). To verify the existence of a difference between the supplemented and the non-supplemented groups, the paired T test was applied, using the Excel / Action software (Microsoft). In vitro embryo production routines supplemented with β-NGF in the maturation medium had, on average, a higher cleavage rate (P = 0.0072) and a higher blastocyst rate (P = 0.0033) compared to non-supplemented routines with β-NGF. In this study was demonstrated that Nerve Growth Factor supplementation in the maturation medium improves the efficiency of in vitro embryo production in cattle, and this protein has a probable action in the oocyte maturation process.

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.

177 PERFORMANCE OF Gyr PREPUBERTAL HEIFERS IN IN VITRO EMBRYO PRODUCTION

2016 ◽  
Vol 28 (2) ◽  
pp. 219
Author(s):  
P. M. S. Rosa ◽  
A. J. R. Camargo ◽  
R. V. Serapião ◽  
L. S. A. Camargo ◽  
C. S. Oliveira
Keyword(s):  
Cleavage Rate ◽  
Embryo Production ◽  
Exact Test ◽  
Follicular Wave ◽  
Lactating Cows ◽  
In Vitro Embryo Production ◽  
Blastocyst Rate ◽  
Oocyte Donors ◽  
La Jolla

Bovine in vitro embryo production is highly relevant for dairy systems in Brazil, and Gyr dams are commonly used as oocyte donors. The aim of this study was to evaluate the use of prepubertal Gyr heifers as oocyte donors, an alternative to anticipate reproduction of those animals. For that, 11 Gyr [4 prepubertal (PP) donors and 7 adult cows © donors] were used in ovum pickup (OPU) sessions. The PP cows presented an average of 282.5 kg and 26.75 months, and had never displayed oestrous. Non-lactating cows presenting an average of 492 kg and 136 months were selected for C. Five replicates were performed, totaling 27 OPU sessions (C-17, PP-10) and 2–3 sessions per animal. Follicular wave was synchronised by aspiration of follicles larger than 8 mm 96 h before OPU. Cumulus-oocyte complexes (COC) were classified accordingly to their quality in viable (G1, G2, and G3) or non-viable (G4). Viable oocytes were matured and fertilized, and the presumptive zygotes were cultured in SOF medium at 38.5°C and 5% CO2 in air. Cleavage rate was assessed 48 to 72 h post-insemination (hpi) and blastocyst rate at 168 hpi. Mean number of structures was analysed by t-test, and percentage of viable, G1, G2, G3, G4, cleavage, and blastocyst rates were compared among groups by Fisher’s exact test (GraphPadInstat, La Jolla, CA, USA; P = 0.05). Results are followed by standard error values. All procedures were approved by a local ethics committee. We found that despite higher (P < 0.05) numbers for both viable oocytes (PP: 15 ± 2.6; C: 6.11 ± 0.76) and total oocytes (PP: 23.70 ± 2.83; C: 8.82 ± 1.19) in the PP group, the rate of viable oocytes was similar (P > 0.05) among PP and C groups (PP: 61.5 ± 6.51%, C: 66.79 ± 3.79%). Mean numbers of G1, G2, G3, and G4 oocytes were higher (P < 0.05) in the PP group (G1 = 7.1 ± 1.18; G2 = 4.9 ± 1.74; G3 = 3.9 ± 1.09; G4 = 7.8 ± 1.38) than in the C group (G1 = 2.70 ± 0.740; G2 = 2.47 ± 0.44; G3 = 1.11 ± 0.31; G4 = 2.52 ± 0.39). However, the proportion was similar (P > 0.05) among PP and C groups (PP: G1 = 29.5 ± 4.21%; G2 = 19.5 ± 2.85%; G3 = 15.9 ± 13.5%; G4 = 35.1 ± 6.33%; and C: G1 = 27.24 ± 4.44%; G2 = 29.60 ± 5.08%; G3 = 12.34 ± 3.01%, G4 = 30.79 ± 4.93%). Cleavage rate (PP: 91.3 ± 17.94%; C: 74.09 ± 4.65%), mean blastocyst number per OPU session (PP: 3.3 ± 1.29; C: 1.76 ± 0.28), and blastocyst rate (PP: 19.74 ± 7.40%; C: 27.03% ± 4.07%) were similar (P > 0.05) among groups. We conclude that prepubertal heifers presented increased numbers of viable oocytes per OPU session, but blastocyst yield was similar to adult cows. This data suggests that prepubertal Gyr heifers can be used as oocyte donors. Support from FAPERJ and Embrapa is acknowledged.

In vitro embryo production from early antral follicles of goats fed with a whole full-fat linseed based diet

Zygote ◽  
2021 ◽  
pp. 1-6
Author(s):  
J. Cadenas ◽  
N.A.R. Sá ◽  
A.C.A. Ferreira ◽  
V.M. Paes ◽  
F.L.N. Aguiar ◽  
...  
Keyword(s):  
Recovery Rate ◽  
Penetration Rate ◽  
Control Treatment ◽  
Follicular Growth ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Feed Supplementation ◽  
Antral Follicles ◽  
Sperm Penetration

Summary The present study aimed to use an in vitro follicle culture (IVFC) biotechnique as a tool to evaluate the influence of whole flaxseed as a feed supplementation in the diet on the in vitro development of caprine early antral follicles (EAFs) and further embryo production. In total, 18 adult goats were homogeneously allocated into two diet groups: Control and Flaxseed. EAFs from both experimental groups (300–400 µm) were isolated and cultured in vitro for 18 days. After IVFC, recovered cumulus–oocyte complexes were submitted to in vitro maturation, and subsequently to IVF and in vitro embryo culture. The endpoints evaluated were follicular growth and morphology, oocyte recovery rate and diameter, sperm penetration, pronuclei formation, embryo development, and estradiol production. The addition of the whole flaxseed in the diet did not affect (P > 0.05) follicular growth and diameter. A higher (P < 0.05) percentage of oocytes ≥ 110 µm was recovered from the flaxseed treatment. However, the sperm penetration rate was higher (P < 0.05) in the control treatment when compared with the flaxseed treatment, but no differences were found regarding the rate of fertilization nor cleaved embryos. In conclusion, dietary flaxseed increased the recovery rate of fully grown oocytes, but it did negatively affect the sperm penetration rate, even though there was no further effect on the cleavage rate.

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